Sample records for zerstoerungsfreie materialpruefung zfp

  1. 2013 annual meeting of the Deutsche Gesellschaft fuer Zerstoerungsfreie Pruefung (DGZfP). NDE in research, development and application; DGZfP-Jahrestagung 2013. Zerstoerungsfreie Materialpruefung. ZfP in Forschung, Entwicklung und Anwendung

    Energy Technology Data Exchange (ETDEWEB)



    This CD-ROM proceedings publication contains the papers and posters of the DGZfP annual meeting 2013, at Bremen, Germany, both in full text and as PowerPoint presentations. The following issues were discussed: computerized tomography, thermography; radiographic, ultrasonic, eddy current and microwave testing. 13 papers were recorded as separate entries in the INIS database.

  2. Diagnosis of structures. Practical applications and future tasks of non-destructive testing; Bauwerksdiagnose. Praktische Anwendungen Zerstoerungsfreier Pruefungen und Zukunftsaufgaben

    Energy Technology Data Exchange (ETDEWEB)



    The preservation of buildings is increasingly becoming the focus of public attention, not least because of traffic restrictions on roads and bridges, which have grown significantly in recent years. Here the building inspectors is of particular importance to assess the causes and extent of repair work as required. This non-destructive testing in the construction industry (ZfPBau method) have become indispensable. However, very few rules are found in construction in contrast to the classical industrial NDT. Also the offers to the qualification of examiners are low, but but in the meantime available. The symposium takes this conflict area to less regulation and a high demand of professional services. One of tasks of the future is the reliable evaluation of existing structures. The foundation was laid by the adoption of the directive for the recalculation of road bridges in 2011 to have results of non-destructive examination incorporated in recalculations. Meanwhile are first experiences on investigations and recalculations of bridges that will be presented at the symposium for the first time. [German] Die Erhaltung von Bauwerken rueckt immer mehr in den Fokus der Oeffentlichkeit, nicht zuletzt durch Verkehrseinschraenkungen an Strassen und Bruecken, die in den letzten Jahren spuerbar zugenommen haben. Dabei kommt den Bauwerkspruefern eine besondere Bedeutung zu, Ursachen und Umfang von Instandsetzungsmassnahmen bedarfsgemaess abzuschaetzen. Hierbei sind zerstoerungsfreie Pruefverfahren im Bauwesen (ZfPBau-Verfahren) nicht mehr wegzudenken. Allerdings sind im Gegensatz zur klassischen industriellen ZfP im Bauwesen sehr wenige Regelwerke anzutreffen. Auch die Angebote zur Qualifizierung von Pruefern sind gering aber mittlerweile vorhanden. Die Fachtagung greift dieses Spannungsfeld geringer Regelsetzung und grossem Bedarf qualifizierter Dienstleistungen auf. Ein Aufgabenfeld der Zukunft ist die zuverlaessige Bewertung von Bestandsbauwerken. Durch das Inkrafttreten der

  3. ZFP36L1 and ZFP36L2 inhibit cell proliferation in a cyclin D-dependent and p53-independent manner


    Suk, Fat-Moon; Chang, Chi-Ching; Lin, Ren-Jye; Lin, Shyr-Yi; Liu, Shih-Chen; Jau, Chia-Feng; Liang, Yu-Chih


    ZFP36 family members include ZFP36, ZFP36L1, and ZFP36L2, which belong to CCCH-type zinc finger proteins with two tandem zinc finger (TZF) regions. Whether ZFP36L1 and ZFP36L2 have antiproliferative activities similar to that of ZFP36 is unclear. In this study, when ZFP36L1 or ZFP36L2 was overexpressed in T-REx-293 cells, cell proliferation was dramatically inhibited and the cell cycle was arrested at the G1 phase. The levels of cell-cycle-related proteins, including cyclin B, cyclin D, cycli...

  4. RNA-binding proteins ZFP36L1 and ZFP36L2 promote cell quiescence. (United States)

    Galloway, Alison; Saveliev, Alexander; Łukasiak, Sebastian; Hodson, Daniel J; Bolland, Daniel; Balmanno, Kathryn; Ahlfors, Helena; Monzón-Casanova, Elisa; Mannurita, Sara Ciullini; Bell, Lewis S; Andrews, Simon; Díaz-Muñoz, Manuel D; Cook, Simon J; Corcoran, Anne; Turner, Martin


    Progression through the stages of lymphocyte development requires coordination of the cell cycle. Such coordination ensures genomic integrity while cells somatically rearrange their antigen receptor genes [in a process called variable-diversity-joining (VDJ) recombination] and, upon successful rearrangement, expands the pools of progenitor lymphocytes. Here we show that in developing B lymphocytes, the RNA-binding proteins (RBPs) ZFP36L1 and ZFP36L2 are critical for maintaining quiescence before precursor B cell receptor (pre-BCR) expression and for reestablishing quiescence after pre-BCR-induced expansion. These RBPs suppress an evolutionarily conserved posttranscriptional regulon consisting of messenger RNAs whose protein products cooperatively promote transition into the S phase of the cell cycle. This mechanism promotes VDJ recombination and effective selection of cells expressing immunoglobulin-μ at the pre-BCR checkpoint. Copyright © 2016, American Association for the Advancement of Science.

  5. Nondestructive characterization of materials damage; Zerstoerungsfreie Charakterisierung von Werkstoffschaedigung

    Energy Technology Data Exchange (ETDEWEB)

    Dobmann, G. [Fraunhofer Inst. fuer Zerstoerungsfreie Pruefverfahren, IZFP, Saarbruecken (Germany)


    The paper discusses two examples of application of NDT for early detection of materials damage, or ageing effects. The first case shows the potential of micromagnetic testing techniques for early detection of hydrogen-induced stress corrosion cracking in pipework. Pipe specimens made of steel X20Cr13, which differed in materials state induced by various heat treatments, were exposed to corrosive attack induced in a closed cycle by simultaneous rinsing with NACE solution and stress application through stamp impact. Various micromagnetic testing parameters were measured at the specimens` outside surface during the annealing period. Further, fatigue experiments were made with specimen sets made of steel 1.4541, under various mechanical and temperature stresses (room temperature and 300 C), for measuring the remanent magnetization of the cylindrical fatigue test blocks (equal geometry) cut out of the specimens, in order to conclude information on the martensite content and the fatigue effects. (Orig./CB) [Deutsch] In diesem Beitrag werden zwei Beispiele der ZFP zur Frueherkennung von Werkstoffschaedigung und Alterungserscheinungen diskutiert. Zum einen wurde das Potential mikromagnetischer Pruefverfahren zur Frueherkennung wasserstoffinduzierter, auch kathodische Spannungsrisskorrosion genannt (HISCC, hydrogene induced stress corrosion cracking), an Rohrleitungen nachgewiesen. Rohrproben unterschiedlicher Waermebehandlungszustaende der Stahlguete X20Cr13 wurden durch gleichzeitiges Spuelen mit NACE-Loesung in einem geschlossenen Kreislauf und Beaufschlagen einer Spannung durch Stempeldruck dem Korrosionsangriff ausgesetzt. Waehrend der Auslagerung wurden verschiedene mikromagnetische Pruefgroessen an der Aussenseite der Rohrproben aufgenommen. Zum anderen wurden an Probensaetzen der Stahlqualitaet 1.4541 Ermuedungsexperimente bei unterschiedlichen mechanischen und Temperaturbelastungen (bei Raumtemperatur und 300 C) durchgefuehrt. Das Ziel war die Messung der

  6. Zfp296 is a novel, pluripotent-specific reprogramming factor.

    Directory of Open Access Journals (Sweden)

    Gerrit Fischedick

    Full Text Available Expression of the four transcription factors Oct4, Sox2, Klf4, and c-Myc (OSKM is sufficient to reprogram somatic cells into induced pluripotent stem (iPSCs. However, this process is slow and inefficient compared with the fusion of somatic cells with embryonic stem cells (ESCs, indicating that ESCs express additional factors that can enhance the efficiency of reprogramming. We had previously developed a method to detect and isolate early neural induction intermediates during the differentiation of mouse ESCs. Using the gene expression profiles of these intermediates, we identified 23 ESC-specific transcripts and tested each for the ability to enhance iPSC formation. Of the tested factors, zinc finger protein 296 (Zfp296 led to the largest increase in mouse iPSC formation. We confirmed that Zfp296 was specifically expressed in pluripotent stem cells and germ cells. Zfp296 in combination with OSKM induced iPSC formation earlier and more efficiently than OSKM alone. Through mouse chimera and teratoma formation, we demonstrated that the resultant iPSCs were pluripotent. We showed that Zfp296 activates transcription of the Oct4 gene via the germ cell-specific conserved region 4 (CR4, and when overexpressed in mouse ESCs leads to upregulation of Nanog expression and downregulation of the expression of differentiation markers, including Sox17, Eomes, and T, which is consistent with the observation that Zfp296 enhances the efficiency of reprogramming. In contrast, knockdown of Zfp296 in ESCs leads to the expression of differentiation markers. Finally, we demonstrated that expression of Zfp296 in ESCs inhibits, but does not block, differentiation into neural cells.

  7. Zfp206 regulates ES cell gene expression and differentiation. (United States)

    Zhang, Wen; Walker, Emily; Tamplin, Owen J; Rossant, Janet; Stanford, William L; Hughes, Timothy R


    Understanding transcriptional regulation in early developmental stages is fundamental to understanding mammalian development and embryonic stem (ES) cell properties. Expression surveys suggest that the putative SCAN-Zinc finger transcription factor Zfp206 is expressed specifically in ES cells [Zhang,W., Morris,Q.D., Chang,R., Shai,O., Bakowski,M.A., Mitsakakis,N., Mohammad,N., Robinson,M.D., Zirngibl,R., Somogyi,E. et al., (2004) J. Biol., 3, 21; Brandenberger,R., Wei,H., Zhang,S., Lei,S., Murage,J., Fisk,G.J., Li,Y., Xu,C., Fang,R., Guegler,K. et al., (2004) Nat. Biotechnol., 22, 707-716]. Here, we confirm this observation, and we show that ZFP206 expression decreases rapidly upon differentiation of cultured mouse ES cells, and during development of mouse embryos. We find that there are at least six isoforms of the ZFP206 transcript, the longest being predominant. Overexpression and depletion experiments show that Zfp206 promotes formation of undifferentiated ES cell clones, and positively regulates abundance of a very small set of transcripts whose expression is also specific to ES cells and the two- to four-cell stages of preimplantation embryos. This set includes members of the Zscan4, Thoc4, Tcstv1 and eIF-1A gene families, none of which have been functionally characterized in vivo but whose members include apparent transcription factors, RNA-binding proteins and translation factors. Together, these data indicate that Zfp206 is a regulator of ES cell differentiation that controls a set of genes expressed very early in development, most of which themselves appear to be regulators.

  8. Structural analysis and dimerization profile of the SCAN domain of the pluripotency factor Zfp206

    KAUST Repository

    Liang, Yu; Huimei Hong, Felicia; Ganesan, Pugalenthi; Jiang, Sizun; Jauch, Ralf; Stanton, Lawrence W.; Kolatkar, Prasanna R.


    Zfp206 (also named as Zscan10) belongs to the subfamily of C2H2 zinc finger transcription factors, which is characterized by the N-terminal SCAN domain. The SCAN domain mediates self-association and association between the members of SCAN family transcription factors, but the structural basis and selectivity determinants for complex formation is unknown. Zfp206 is important for maintaining the pluripotency of embryonic stem cells presumably by combinatorial assembly of itself or other SCAN family members on enhancer regions. To gain insights into the folding topology and selectivity determinants for SCAN dimerization, we solved the 1.85 crystal structure of the SCAN domain of Zfp206. In vitro binding studies using a panel of 20 SCAN proteins indicate that the SCAN domain Zfp206 can selectively associate with other members of SCAN family transcription factors. Deletion mutations showed that the N-terminal helix 1 is critical for heterodimerization. Double mutations and multiple mutations based on the Zfp206SCAN-Zfp110SCAN model suggested that domain swapped topology is a possible preference for Zfp206SCAN-Zfp110SCAN heterodimer. Together, we demonstrate that the Zfp206SCAN constitutes a protein module that enables C2H2 transcription factor dimerization in a highly selective manner using a domain-swapped interface architecture and identify novel partners for Zfp206 during embryonal development. 2012 The Author(s).

  9. Structural analysis and dimerization profile of the SCAN domain of the pluripotency factor Zfp206

    KAUST Repository

    Liang, Yu


    Zfp206 (also named as Zscan10) belongs to the subfamily of C2H2 zinc finger transcription factors, which is characterized by the N-terminal SCAN domain. The SCAN domain mediates self-association and association between the members of SCAN family transcription factors, but the structural basis and selectivity determinants for complex formation is unknown. Zfp206 is important for maintaining the pluripotency of embryonic stem cells presumably by combinatorial assembly of itself or other SCAN family members on enhancer regions. To gain insights into the folding topology and selectivity determinants for SCAN dimerization, we solved the 1.85 crystal structure of the SCAN domain of Zfp206. In vitro binding studies using a panel of 20 SCAN proteins indicate that the SCAN domain Zfp206 can selectively associate with other members of SCAN family transcription factors. Deletion mutations showed that the N-terminal helix 1 is critical for heterodimerization. Double mutations and multiple mutations based on the Zfp206SCAN-Zfp110SCAN model suggested that domain swapped topology is a possible preference for Zfp206SCAN-Zfp110SCAN heterodimer. Together, we demonstrate that the Zfp206SCAN constitutes a protein module that enables C2H2 transcription factor dimerization in a highly selective manner using a domain-swapped interface architecture and identify novel partners for Zfp206 during embryonal development. 2012 The Author(s).

  10. Zfp462 deficiency causes anxiety-like behaviors with excessive self-grooming in mice. (United States)

    Wang, B; Zheng, Y; Shi, H; Du, X; Zhang, Y; Wei, B; Luo, M; Wang, H; Wu, X; Hua, X; Sun, M; Xu, X


    Zfp462 is a newly identified vertebrate-specific zinc finger protein that contains nearly 2500 amino acids and 23 putative C2H2-type zinc finger domains. So far, the functions of Zfp462 remain unclear. In our study, we showed that Zfp462 is expressed predominantly in the developing brain, especially in the cerebral cortex and hippocampus regions from embryonic day 7.5 to early postnatal stage. By using a piggyBac transposon-generated Zfp462 knockout (KO) mouse model, we found that Zfp462 KO mice exhibited prenatal lethality with normal neural tube patterning, whereas heterozygous (Het) Zfp462 KO (Zfp462 +/- ) mice showed developmental delay with low body weight and brain weight. Behavioral studies showed that Zfp462 +/- mice presented anxiety-like behaviors with excessive self-grooming and hair loss, which were similar to the pathological grooming behaviors in Hoxb8 KO mice. Further analysis of grooming microstructure showed the impairment of grooming patterning in Zfp462 +/- mice. In addition, the mRNA levels of Pbx1 (pre-B-cell leukemia homeobox 1, an interacting protein of Zfp462) and Hoxb8 decreased in the brains of Zfp462 +/- mice, which may be the cause of anxiety-like behaviors. Finally, imipramine, a widely used and effective anti-anxiety medicine, rescued anxiety-like behaviors and excessive self-grooming in Zfp462 +/- mice. In conclusion, Zfp462 deficiency causes anxiety-like behaviors with excessive self-grooming in mice. This provides a novel genetic mouse model for anxiety disorders and a useful tool to determine potential therapeutic targets for anxiety disorders and screen anti-anxiety drugs. © 2016 John Wiley & Sons Ltd and International Behavioural and Neural Genetics Society.

  11. The RNA-binding protein, ZFP36L2, influences ovulation and oocyte maturation.

    Directory of Open Access Journals (Sweden)

    Christopher B Ball

    Full Text Available ZFP36L2 protein destabilizes AU-rich element-containing transcripts and has been implicated in female fertility. In the C57BL/6NTac mouse, a mutation in Zfp36l2 that results in the decreased expression of a form of ZFP36L2 in which the 29 N-terminal amino acid residues have been deleted, ΔN-ZFP36L2, leads to fertilized eggs that arrest at the two-cell stage. Interestingly, homozygous ΔN-Zfp36l2 females in the C57BL/6NTac strain release 40% fewer eggs than the WT littermates (Ramos et al., 2004, suggesting an additional defect in ovulation and/or oocyte maturation. Curiously, the same ΔN-Zfp36l2 mutation into the SV129 strain resulted in anovulation, prompting us to investigate a potential problem in ovulation and oocyte maturation. Remarkably, only 20% of ΔN-Zfp36l2 oocytes in the 129S6/SvEvTac strain matured ex vivo, suggesting a defect on the oocyte meiotic maturation process. Treatment of ΔN-Zfp36l2 oocytes with a PKA inhibitor partially rescued the meiotic arrested oocytes. Furthermore, cAMP levels were increased in ΔN-Zfp36l2 oocytes, linking the cAMP/PKA pathway and ΔN-Zfp36l2 with meiotic arrest. Since ovulation and oocyte maturation are both triggered by LHR signaling, the downstream pathway was investigated. Adenylyl cyclase activity was increased in ΔN-Zfp36l2 ovaries only upon LH stimulation. Moreover, we discovered that ZFP36L2 interacts with the 3'UTR of LHR mRNA and that decreased expression levels of Zfp36l2 correlates with higher levels of LHR mRNA in synchronized ovaries. Furthermore, overexpression of ZFP36L2 decreases the endogenous expression of LHR mRNA in a cell line. Therefore, we propose that lack of the physiological down regulation of LHR mRNA levels by ZFP36L2 in the ovaries is associated with anovulation and oocyte meiotic arrest.

  12. The centromeric/nucleolar chromatin protein ZFP-37 may function to specify neuronal nuclear domains

    NARCIS (Netherlands)

    E. Payen; T. Verkerk (Ton); D. Michalovich (David); S.D. Dreyer; A. Winterpacht; B. Lee (Brendan); C.I. de Zeeuw (Chris); N.J. Galjart (Niels); F.G. Grosveld (Frank)


    textabstractMurine ZFP-37 is a member of the large family of C2H2 type zinc finger proteins. It is characterized by a truncated NH2-terminal Kruppel-associated box and is thought to play a role in transcriptional regulation. During development Zfp-37 mRNA is most

  13. 2003 annual meeting of the Deutsche Gesellschaft fuer zerstoerungsfreie Pruefung (DGZfP): NDE in research, development and application

    International Nuclear Information System (INIS)


    The annual meeting 2004 of the Deutsche Gesellschaft fuer Zerstoerungsfreie Pruefung e.V. (Berlin, Federal Republic of Germany) was proceeded in Salzburg (Austria) between 17th and 19th May, 2004. The topics of this proceeding are in general all aspects of non-destructive materials testing: repeating tests/inspections, characterization of materials, aerospace, simulation, automobiles, metallic products, optical processes, testing of tubes, testing of acoustic emission, digital radiography, technology of ultrasonic devices, testing of containers, railway, standardization and standards; computer tomography; thermography; special ultrasonic technologies; ultrasonic testing; transmission testing; eddy current testing; surface processes; radiation protection; construction industry; quality assurance

  14. 2007 annual meeting of the Deutsche Gesellschaft fuer Zerstoerungsfreie Pruefung (DGZfP): NDE in research, development and application

    International Nuclear Information System (INIS)


    The 2007 annual meeting of Deutsche Gesellschaft fuer Zerstoerungsfreie Pruefung was held at Fuerth in northern Bavaria, a town that had its 1000th anniversary in 2007. Subjects discussed at the conference were new materials, modern technologies, and innovative non-destructive testing. The conference was held close to the ''Technikum Fuerth'' and ''Uferstadt Fuerth'', both of which are centers of research and seats of young, innovative businesses, which gave young scientists an opportunity to present the results of their work and allowed the DGZfPL to present itself to young scientists and to invite discussions. (orig.)

  15. ZFPIP/Zfp462 is involved in P19 cell pluripotency and in their neuronal fate

    International Nuclear Information System (INIS)

    Masse, Julie; Piquet-Pellorce, Claire; Viet, Justine; Guerrier, Daniel; Pellerin, Isabelle; Deschamps, Stephane


    The nuclear zinc finger protein ZFPIP/Zfp462 is an important factor involved in cell division during the early embryonic development of vertebrates. In pluripotent P19 cells, ZFPIP/Zfp462 takes part in cell proliferation, likely via its role in maintaining chromatin structure. To further define the function of ZFPIP/Zfp462 in the mechanisms of pluripotency and cell differentiation, we constructed a stable P19 cell line in which ZFPIP/Zfp462 knockdown is inducible. We report that ZFPIP/Zfp462 was vital for mitosis and self-renewal in pluripotent P19 cells. Its depletion induced substantial decreases in the expression of the pluripotency genes Nanog, Oct4 and Sox2 and was associated with the transient expression of specific neuronal differentiation markers. We also demonstrated that ZFPIP/Zfp462 expression appears to be unnecessary after neuronal differentiation is induced in P19 cells. Taken together, our results strongly suggest that ZFPIP/Zfp462 is a key chromatin factor involved in maintaining P19 pluripotency and in the early mechanisms of neural differentiation but that it is dispensable in differentiated P19 cells.

  16. Molecular characterization and expression of DgZFP1, a gene ...

    African Journals Online (AJOL)



    Apr 12, 2010 ... Full Length Research Paper. Molecular characterization and ... stem mainly done by removing the auxiliary flower buds. But cultivars as potted ... DgZFP1 by using the first strand cDNA of chrysanthemum as a template.

  17. ZFP226 is a novel artificial transcription factor for selective activation of tumor suppressor KIBRA. (United States)

    Schelleckes, Katrin; Schmitz, Boris; Lenders, Malte; Mewes, Mirja; Brand, Stefan-Martin; Brand, Eva


    KIBRA has been suggested as a key regulator of the hippo pathway, regulating organ size, cell contact inhibition as well as tissue regeneration and tumorigenesis. Recently, alterations of KIBRA expression caused by promotor methylation have been reported for several types of cancer. Our current study aimed to design an artificial transcription factor capable of re-activating expression of the tumor suppressor KIBRA and the hippo pathway. We engineered a new gene named 'ZFP226' encoding for a ~23 kDa fusion protein. ZFP226 belongs to the Cys2-His2 zinc finger type and recognizes a nine base-pair DNA sequence 5'-GGC-GGC-GGC-3' in the KIBRA core promoter P1a. ZFP226 showed nuclear localization in human immortalized kidney epithelial cells and activated the KIBRA core promoter (p < 0.001) resulting in significantly increased KIBRA mRNA and protein levels (p < 0.001). Furthermore, ZFP226 led to activation of hippo signaling marked by elevated YAP and LATS phosphorylation. In Annexin V flow cytometry assays ZFP226 overexpression showed strong pro-apoptotic capacity on MCF-7 breast cancer cells (p < 0.01 early-, p < 0.001 late-apoptotic cells). We conclude that the artificial transcription factor ZFP226 can be used for target KIBRA and hippo pathway activation. This novel molecule may represent a molecular tool for the development of future applications in cancer treatment.

  18. Optical spectroscopy and luminescence properties of Ho3+ doped zinc fluorophosphate (ZFP) glasses for green luminescent device applications (United States)

    Reddy Prasad, V.; Damodaraiah, S.; Ratnakaram, Y. C.


    Ho3+ doped zinc fluorophosphate (ZFP) glasses with molar chemical compositions, (60-x) NH4H2PO4+20ZnO+10BaF2+10NaF+xHo2O3 (where x = 0.1, 0.3, 0.5, 1.0 and 1.5 mol%) were prepared by melt quenching technique. These glasses were characterized through physical, structural, optical, excitation, luminescence and decay curve analysis. From the absorption spectra, spectral intensities (fexp and fcal), Judd-Ofelt intensity parameters (Ω2, Ω4 and Ω6), radiative transition probabilities (AT), radiative lifetimes (τR) and branching ratios (βR) were evaluated for all Ho3+ doped ZFP glass matrices. From the photoluminescence spectra, peak stimulated emission cross-sections (σP) were calculated for all Ho3+ doped ZFP glasses. The Ho3+ doped ZFP glasses show strong green emission at 545 nm and red emission at 656 nm under excitation, 450 nm. The measured lifetimes (τmeas) of (5S2)5F4 level of Ho3+ doped ZFP glasses were obtained from decay profiles. The CIE color coordinates of Ho3+ doped ZFP glasses were calculated from emission spectra and 1.0 mol% of Ho3+ doped ZFP glass matrix gives green emission. Hence, these results confirm that the Ho3+ doped ZFP glasses could be considered as a promising candidate for visible green laser applications.

  19. Genetic and epigenetic mutations affect the DNA binding capability of human ZFP57 in transient neonatal diabetes type 1. (United States)

    Baglivo, Ilaria; Esposito, Sabrina; De Cesare, Lucia; Sparago, Angela; Anvar, Zahra; Riso, Vincenzo; Cammisa, Marco; Fattorusso, Roberto; Grimaldi, Giovanna; Riccio, Andrea; Pedone, Paolo V


    In the mouse, ZFP57 contains three classical Cys2His2 zinc finger domains (ZF) and recognizes the methylated TGC(met)CGC target sequence using the first and the second ZFs. In this study, we demonstrate that the human ZFP57 (hZFP57) containing six Cys2His2 ZFs, binds the same methylated sequence through the third and the fourth ZFs, and identify the aminoacids critical for DNA interaction. In addition, we present evidences indicating that hZFP57 mutations and hypomethylation of the TNDM1 ICR both associated with Transient Neonatal Diabetes Mellitus type 1 result in loss of hZFP57 binding to the TNDM1 locus, likely causing PLAGL1 activation. Copyright © 2013 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

  20. No evidence for pathogenic variants or maternal effect of ZFP57 as the cause of Beckwith-Wiedemann Syndrome

    DEFF Research Database (Denmark)

    Boonen, Susanne E; Hahnemann, Johanne M D; Mackay, Deborah


    in patients with BWS. We sequenced ZFP57 in 27 BWS probands and in 23 available mothers to test for a maternal effect. We identified three novel, presumably benign sequence variants in ZFP57; thus, we found no evidence for ZFP57 alterations as a major cause in sporadic BWS cases.......Beckwith-Wiedemann syndrome (BWS) is an overgrowth syndrome, which, in 50-60% of sporadic cases, is caused by hypomethylation of KCNQ1OT1 differentially methylated region (DMR) at chromosome 11p15.5. The underlying defect of this hypomethylation is largely unknown. Recently, recessive mutations...... of the ZFP57 gene were reported in patients with transient neonatal diabetes mellitus type 1, showing hypomethylation at multiple imprinted loci, including KCNQ1OT1 DMR in some. The aim of our study was to determine whether ZFP57 alterations were a genetic cause of the hypomethylation at KCNQ1OT1 DMR...

  1. Crystal optimization and preliminary diffraction data analysis of the SCAN domain of Zfp206

    International Nuclear Information System (INIS)

    Liang, Yu; Choo, Siew Hua; Rossbach, Michael; Baburajendran, Nithya; Palasingam, Paaventhan; Kolatkar, Prasanna R


    Crystals of the SCAN domain of Zfp206 are tetragonal, belonging to space group I422 with unit-cell parameters a = 67.57, c = 87.54 Å and one molecule in the asymmetric unit, and diffract to 1.85 Å resolution. Zfp206 (also named Zscan10) is a transcription factor that plays an important role in maintaining the pluripotent state of embryonic stem cells. Zfp206 is a member of the SCAN-domain family of C 2 H 2 zinc-finger transcription factors. The SCAN domain is a highly conserved motif of 84 residues which mediates the self-association of and heterodimerization between SCAN-domain family transcription factors. The SCAN domain may therefore be the key to the selective oligomerization of and may combinatorially enhance the regulatory versatility of C 2 H 2 zinc fingers. This paper describes crystallization attempts with the SCAN domain of Zfp206 (Zfp206SCAN) and optimization strategies to obtain diffraction-quality crystals. The best diffracting crystal was grown in a solution consisting of 0.3 M ammonium sulfate, 0.1 M Tris–HCl pH 8.6, 25% PEG 3350, 0.1 M ethylenediaminetetraacetic acid disodium salt dehydrate (EDTA) using the hanging-drop vapour-diffusion technique. Optimized crystals diffracted to 1.85 Å resolution and belonged to space group I422, with unit-cell parameters a = 67.57, c = 87.54 Å. A Matthews analysis indicated the presence of one Zfp206SCAN molecule per asymmetric unit

  2. Utilities` view about NDT-trends; Entwicklungstendenzen in der ZfP aus Betreibersicht

    Energy Technology Data Exchange (ETDEWEB)

    Hansch, M.K.T.; Broecker, B. [PreussenElektra AG (Germany)


    Nondestructive testing of nuclear plant components has become an increasingly important part in the performance of ISI activities. A change can be observed over recent years in ISI planning in that there is a trend towards placing more emphasis on component risk informed in-service inspection and online monitoring of components in response to operating history and conditions. Within the framework of the basic safety and in-service inspection concept, NDT and online monitoring of components are redundant, though important measures. This change of approach is a reaction to the advancements in the technical development of NDT methods and capabilities as well as related aspects, such as equipment and techniques for preparation, performance, and evaluation of inspections and subsequent data processing. (orig./CB) [Deutsch] Die zerstoerungsfreie Pruefung in den nuklearen Anlagen ist in den letzten Jahren immer mehr ins Rampenlicht getreten. Die betriebsbedingte Vorgehensweise bei der WKP ist im Rahmen der Betriebsueberwachung der Komponenten eine Tendenz zu einer kausalen wiederkehrenden Pruefung innerhalb des Basissicherheitskonzeptes, das die Betriebsueberwachung und die zerstoerungsfreie Pruefung als redundante Massnahmen involviert. Begleitet wird das o.g. Vorgehen von der technischen Entwicklung der zerstoerungsfreien Prueftechniken und deren Randgebiete, die die Vorbereitung, die Durchfuehrung, die Bewertung und die weitere Datenverwendung betreffen. (orig.)

  3. Transient neonatal diabetes, ZFP57, and hypomethylation of multiple imprinted loci

    DEFF Research Database (Denmark)

    Boonen, Susanne E; Mackay, Deborah J G; Hahnemann, Johanne M D


    Transient neonatal diabetes mellitus 1 (TNDM1) is the most common cause of diabetes presenting at birth. Approximately 5% of the cases are due to recessive ZFP57 mutations, causing hypomethylation at the TNDM locus and other imprinted loci (HIL). This has consequences for patient care because...

  4. Involvement of ZFPIP/Zfp462 in chromatin integrity and survival of P19 pluripotent cells

    International Nuclear Information System (INIS)

    Masse, Julie; Laurent, Audrey; Nicol, Barbara; Guerrier, Daniel; Pellerin, Isabelle; Deschamps, Stephane


    Toti- or pluripotent cells proliferation and/or differentiation have been shown to be strongly related to nuclear chromatin organization and structure over the last past years. We have recently identified ZFPIP/Zfp462 as a zinc finger nuclear factor necessary for correct cell division during early embryonic developmental steps of vertebrates. We thus questioned whether this factor was playing a general role during cell division or if it was somehow involved in embryonic cell fate or differentiation. To achieve this goal, we performed a knock-down experiment in the pluripotent P19 and differentiated 3T3 cell lines, both expressing endogenous ZFPIP/Zfp462. Using specific shRNA directed against ZFPIP/Zfp462 transcripts, we demonstrated that depletion of this protein induced cell death in P19 but had no effect in 3T3 cells. In addition, in the absence of the protein, the P19 cells exhibited a complete destructuration of pericentromeric domains associated with a redistribution of the HP1α proteins and an increase in DNA satellites transcribed RNAs level. These data suggested an instrumental role of ZFPIP/Zfp462 in maintaining the chromatin structure of pluripotent cells.

  5. Expert`s dictionary of technical terminology in non-destructive materials testing. 1359 terms and definitions from A - Z used in non-destructive materials and workpiece testing - for vocational training, students and professionals; Expert-Praxislexikon Zerstoerungsfreie Materialpruefung. 1359 Begriffe von A bis Z zur zerstoerungsfreien Material- und Werkstueckuntersuchung - fuer Lehre, Studium und Beruf

    Energy Technology Data Exchange (ETDEWEB)

    Steeb, S


    The monolingual, German dictionary is a compilation enhanced by figures and tables, of technical terms and definitions used in non-destructive materials testing and application of the following eight major testing methods: - Radiography using X-rays - radiography using isotope beams - crack examination with the magnetic particle method - magnetic stray field testing with scanning probe - penetration tests with light and uv radiation - ultrasonic tests including evaluation of results - eddy current testing, manual and automated - sound emission methods. (orig./MM) 179 figs., 8 tabs. [Deutsch] Die wichtigsten Begriffe aus dem Gebiet der folgenden acht Untersuchungsverfahren sind in diesem Woerterbuch zusammengestellt und erlaeutert: - Durchstrahlungsverfahren mit Roentgenstrahlen - Durchstrahlungsverfahren mit Isotopenstrahlen - Risspruefung nach dem Magnetpulververfahren - magnetische Streuflusspruefung mit Sondenabtastung - Eindringverfahren bei sichtbarem Licht und bei UV-Licht - Ultraschallverfahren mit Bewertung der Ergebnisse - Wirbelstromverfahren manuell und automatisch - Schallemissionsverfahren. (orig./MM)

  6. Diagnosis of structures. Practical applications and future tasks of non-destructive testing. Papers; Bauwerksdiagnose. Praktische Anwendungen Zerstoerungsfreier Pruefungen und Zukunftsaufgaben. Vortraege

    Energy Technology Data Exchange (ETDEWEB)



    The CD contains 17 papers and 20 posters on the subject of non-destructive diagnosis of structures. The titles of the 17 papers are: 1. Non-destructive testing in transition - from quality testing to life management; 2. Non-destructive testing of bridges on German motorways; 3. Structural diagnosis of concrete components - results obtained by a group of scientists of Deutsche Forschungsgemeinschaft; 4. Building diagnosis - its relevance and options for the building authorities; 5. Optimisation of permanent monitoring of structures by non-destructive test methods; 6. Assessment of Structures using Fibre-optic Sensors; 7. Methods and results in wood structure diagnosis; 8. The collapse of the roof of the Bad Reichenhall ice-skating rink - how could it have been prevented? 9. Integration of non-destructive structural tests in structure management of PPP models (Powerpoint); 10: Elements of sustainable life cycle management of engineering structures; 11. Structure monitoring as a component of life management systems; 12. Chloride and Sulfate analysis using LIBS - from the laboratory to the construction site; 13. Applications of georadar und ultrasonics - examples from the working practice of an engineering consultants' office; 14. Bridge analysis by automatic methods - new developments; 15. Fast large-area condition monitoring of reinforced-concrete bridges, floors and ceilings; 16. Influence of roof gap pressing on non-destructive thickness testing of the inner shells of motorway tunnels; 17. Non-destructive test methods for structure diagnosis - status and perspectives. [German] Die CD enthaelt 17 Vortraege und 20 Posterbeitraege zum Thema: ''Zerstoerungsfreie Pruefverahren fuer die Bauwerksdiagnose''. Die Themen der 17 Vortraege sind: 1. Die Zerstoerungsfreie Pruefung im Wandel - Von der Qualitaetspruefung zum Lebensdauermanagement; 2. ZfPBau-Verfahren fuer die Zustandserfassung von Bruecken der Bundesfernstrassen; 3. Strukturbestimmung von

  7. Neuronal migration is regulated by endogenous RNAi and chromatin-binding factor ZFP-1/AF10 in Caenorhabditis elegans. (United States)

    Kennedy, Lisa M; Grishok, Alla


    Endogenous short RNAs and the conserved plant homeodomain (PHD) zinc-finger protein ZFP-1/AF10 regulate overlapping sets of genes in Caenorhabditis elegans, which suggests that they control common biological pathways. We have shown recently that the RNAi factor RDE-4 and ZFP-1 negatively modulate transcription of the insulin/PI3 signaling-dependent kinase PDK-1 to promote C. elegans fitness. Moreover, we have demonstrated that the insulin/IGF-1-PI3K-signaling pathway regulates the activity of the DAF-16/FOXO transcription factor in the hypodermis to nonautonomously promote the anterior migrations of the hermaphrodite-specific neurons (HSNs) during embryogenesis of C. elegans. In this study, we implicate the PHD-containing isoform of ZFP-1 and endogenous RNAi in the regulation of HSN migration. ZFP-1 affects HSN migration in part through its negative effect on pdk-1 transcription and modulation of downstream DAF-16 activity. We also identify a novel role for ZFP-1 and RNAi pathway components, including RDE-4, in the regulation of HSN migration in parallel with DAF-16. Therefore, the coordinated activities of DAF-16, ZFP-1, and endogenous RNAi contribute to gene regulation during development to ensure proper neuronal positioning.

  8. ZFP521 regulates murine hematopoietic stem cell function and facilitates MLL-AF9 leukemogenesis in mouse and human cells. (United States)

    Garrison, Brian S; Rybak, Adrian P; Beerman, Isabel; Heesters, Balthasar; Mercier, Francois E; Scadden, David T; Bryder, David; Baron, Roland; Rossi, Derrick J


    The concept that tumor-initiating cells can co-opt the self-renewal program of endogenous stem cells as a means of enforcing their unlimited proliferative potential is widely accepted, yet identification of specific factors that regulate self-renewal of normal and cancer stem cells remains limited. Using a comparative transcriptomic approach, we identify ZNF521 / Zfp521 as a conserved hematopoietic stem cell (HSC)-enriched transcription factor in human and murine hematopoiesis whose function in HSC biology remains elusive. Competitive serial transplantation assays using Zfp521 -deficient mice revealed that ZFP521 regulates HSC self-renewal and differentiation. In contrast, ectopic expression of ZFP521 in HSCs led to a robust maintenance of progenitor activity in vitro. Transcriptional analysis of human acute myeloid leukemia (AML) patient samples revealed that ZNF521 is highly and specifically upregulated in AMLs with MLL translocations. Using an MLL-AF9 murine leukemia model and serial transplantation studies, we show that ZFP521 is not required for leukemogenesis, although its absence leads to a significant delay in leukemia onset. Furthermore, knockdown of ZNF521 reduced proliferation in human leukemia cell lines possessing MLL-AF9 translocations. Taken together, these results identify ZNF521/ZFP521 as a critical regulator of HSC function, which facilitates MLL-AF9-mediated leukemic disease in mice.

  9. ZFP57 maintains the parent-of-origin-specific expression of the imprinted genes and differentially affects non-imprinted targets in mouse embryonic stem cells

    DEFF Research Database (Denmark)

    Riso, Vincenzo; Cammisa, Marco; Kukreja, Harpreet


    ZFP57 is necessary for maintaining repressive epigenetic modifications at Imprinting control regions (ICRs). In mouse embryonic stem cells (ESCs), ZFP57 binds ICRs (ICRBS) and many other loci (non-ICRBS). To address the role of ZFP57 on all its target sites, we performed high-throughput and multi......-locus analyses of inbred and hybrid mouse ESC lines carrying different gene knockouts. By using an allele-specific RNA-seq approach, we demonstrate that ZFP57 loss results in derepression of the imprinted allele of multiple genes in the imprinted clusters. We also find marked epigenetic differences between ICRBS...... the imprinted expression over long distances. At non-ICRBS, ZFP57 inactivation results in acquisition of epigenetic features that are characteristic of poised enhancers, suggesting that another function of ZFP57 in early embryogenesis is to repress cis-acting regulatory elements whose activity is not yet...

  10. ThDof1.4 and ThZFP1 constitute a transcriptional regulatory cascade involved in salt or osmotic stress in Tamarix hispida. (United States)

    Zang, Dandan; Wang, Lina; Zhang, Yiming; Zhao, Huimin; Wang, Yucheng


    Identification of the upstream regulators of a gene is important to characterize the transcriptional pathway and the function of the gene. Previously, we found that a zinc finger protein (ThZFP1) is involved in abiotic stress tolerance of Tamarix hispida. In the present study, we further investigated the transcriptional pathway of ThZFP1. Dof motifs are abundant in the ThZFP1 promoter; therefore, we used them to screen for transcriptional regulators of ThZFP1. A Dof protein, ThDof1.4, binds to the Dof motif specifically, and was hypothesized as the upstream regulator of ThZFP1. Further study showed that overexpression of ThDof1.4 in T. hispida activated the expression of GUS controlled by the ThZFP1 promoter. In T. hispida, transient overexpression of ThDof1.4 increased the transcripts of ThZFP1; conversely, transient RNAi-silencing of ThDof1.4 reduced the expression of ThZFP1. Chromatin immunoprecipitation indicated that ThDof1.4 binds to the ThZFP1 promoter. Additionally, ThDof1.4 and ThZFP1 share similar expression patterns in response to salt or drought stress. Furthermore, like ThZFP1, ThDof1.4 could increase the proline level and enhance ROS scavenging capability to improve salt and osmotic stress tolerance. Together, these results suggested that ThDof1.4 and ThZFP1 form a transcriptional regulatory cascade involved in abiotic stress resistance in T. hispida.

  11. Tamarix hispida zinc finger protein ThZFP1 participates in salt and osmotic stress tolerance by increasing proline content and SOD and POD activities. (United States)

    Zang, Dandan; Wang, Chao; Ji, Xiaoyu; Wang, Yucheng


    Zinc finger proteins (ZFPs) are a large family that play important roles in various biological processes, such as signal transduction, RNA binding, morphogenesis, transcriptional regulation, abiotic or biotic stress response. However, the functions of ZFPs involved in abiotic stress are largely not known. In the present study, we cloned and functionally characterized a ZFP gene, ThZFP1, from Tamarix hispida. The expression of ThZFP1 is highly induced by NaCl, mannitol or ABA treatment. To study the function of ThZFP1 involved in abiotic stress response, transgenic T. hispida plants with overexpression or knockdown of ThZFP1 were generated using a transient transformation system. Gain- and loss-of-function studies of ThZFP1 suggested that ThZFP1 can induce the expression of a series of genes, including delta-pyrroline-5-carboxylate synthetase (P5CS), peroxidase (POD) and superoxide dismutase (SOD), leading to accumulation of proline and enhanced activities of SOD and POD. These physiological changes enhanced proline content and reactive oxygen species (ROS) scavenging capability when exposed to salt or osmotic stress. All the results obtained from T. hispida plants were further confirmed by analyses of the transgenic Arabidopsis plants overexpressing ThZFP1. These data together suggested that ThZFP1 positively regulates proline accumulation and activities of SOD and POD under salt and osmotic stress conditions. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  12. Overexpression of GsZFP1 enhances salt and drought tolerance in transgenic alfalfa (Medicago sativa L.). (United States)

    Tang, Lili; Cai, Hua; Ji, Wei; Luo, Xiao; Wang, Zhenyu; Wu, Jing; Wang, Xuedong; Cui, Lin; Wang, Yang; Zhu, Yanming; Bai, Xi


    GsZFP1 encodes a Cys2/His2-type zinc-finger protein. In our previous study, when GsZFP1 was heterologously expressed in Arabidopsis, the transgenic Arabidopsis plants exhibited enhanced drought and cold tolerance. However, it is still unknown whether GsZFP1 is also involved in salt stress. GsZFP1 is from the wild legume Glycine soja. Therefore, the aims of this study were to further elucidate the functions of the GsZFP1 gene under salt and drought stress in the forage legume alfalfa and to investigate its biochemical and physiological functions under these stress conditions. Our data showed that overexpression of GsZFP1 in alfalfa resulted in enhanced salt tolerance. Under high salinity stress, greater relative membrane permeability and malondialdehyde (MDA) content were observed and more free proline and soluble sugars accumulated in transgenic alfalfa than in the wild-type (WT) plants; in addition, the transgenic lines accumulated less Na(+) and more K(+) in both the shoots and roots. Overexpression of GsZFP1 also enhanced the drought tolerance of alfalfa. The fold-inductions of stress-responsive marker gene expression, including MtCOR47, MtRAB18, MtP5CS, and MtRD2, were greater in transgenic alfalfa than those of WT under drought stress conditions. In conclusion, the transgenic alfalfa plants generated in this study could be used for farming in salt-affected as well as arid and semi-arid areas. Copyright © 2013 Elsevier Masson SAS. All rights reserved.

  13. Striking a balance: regulation of transposable elements by Zfp281 and Mll2 in mouse embryonic stem cells. (United States)

    Dai, Qian; Shen, Yang; Wang, Yan; Wang, Xin; Francisco, Joel Celio; Luo, Zhuojuan; Lin, Chengqi


    Transposable elements (TEs) compose about 40% of the murine genome. Retrotransposition of active TEs such as LINE-1 (L1) tremendously impacts genetic diversification and genome stability. Therefore, transcription and transposition activities of retrotransposons are tightly controlled. Here, we show that the Krüppel-like zinc finger protein Zfp281 directly binds and suppresses a subset of retrotransposons, including the active young L1 repeat elements, in mouse embryonic stem (ES) cells. In addition, we find that Zfp281-regulated L1s are highly enriched for 5-hydroxymethylcytosine (5hmC) and H3K4me3. The COMPASS-like H3K4 methyltransferase Mll2 is the major H3K4me3 methylase at the Zfp281-regulated L1s and required for their proper expression. Our studies also reveal that Zfp281 functions partially through recruiting the L1 regulators DNA hydroxymethylase Tet1 and Sin3A, and restricting Mll2 at these active L1s, leading to their balanced expression. In summary, our data indicate an instrumental role of Zfp281 in suppressing the young active L1s in mouse ES cells. © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.

  14. Maintenance of the marginal-zone B cell compartment specifically requires the RNA-binding protein ZFP36L1. (United States)

    Newman, Rebecca; Ahlfors, Helena; Saveliev, Alexander; Galloway, Alison; Hodson, Daniel J; Williams, Robert; Besra, Gurdyal S; Cook, Charlotte N; Cunningham, Adam F; Bell, Sarah E; Turner, Martin


    RNA-binding proteins of the ZFP36 family are best known for inhibiting the expression of cytokines through binding to AU-rich elements in the 3' untranslated region and promoting mRNA decay. Here we identified an indispensable role for ZFP36L1 as the regulator of a post-transcriptional hub that determined the identity of marginal-zone B cells by promoting their proper localization and survival. ZFP36L1 controlled a gene-expression program related to signaling, cell adhesion and locomotion; it achieved this in part by limiting expression of the transcription factors KLF2 and IRF8, which are known to enforce the follicular B cell phenotype. These mechanisms emphasize the importance of integrating transcriptional and post-transcriptional processes by RNA-binding proteins for maintaining cellular identity among closely related cell types.

  15. Maintenance of the marginal zone B cell compartment specifically requires the RNA-binding protein ZFP36L1 (United States)

    Newman, Rebecca; Ahlfors, Helena; Saveliev, Alexander; Galloway, Alison; Hodson, Daniel J; Williams, Robert; Besra, Gurdyal S.; Cook, Charlotte N; Cunningham, Adam F; Bell, Sarah E; Turner, Martin


    RNA binding proteins (RBP) of the ZFP36 family are best known for inhibiting the expression of cytokines through binding to AU rich elements in the 3’UTR and promoting mRNA decay. Here we show an indispensible role for ZFP36L1 as the regulator of a post-transcriptional hub that determined the identity of marginal zone (MZ) B cells by promoting their proper localization and survival. ZFP36L1 controlled a gene expression program related to signaling, cell-adhesion and locomotion, in part by limiting the expression of the transcription factors KLF2 and IRF8, which are known to enforce the follicular B cell phenotype. These mechanisms emphasize the importance of integrating transcriptional and post-transcriptional processes by RBP for maintaining cellular identity between closely related cell types. PMID:28394372

  16. Manipulation of flowering time and branching by overexpression of the tomato transcription factor SlZFP2. (United States)

    Weng, Lin; Bai, Xiaodong; Zhao, Fangfang; Li, Rong; Xiao, Han


    Flowering of higher plants is orchestrated by complex regulatory networks through integration of various environmental signals such as photoperiod, temperature, light quality and developmental cues. In Arabidopsis, transcription of the flowering integrator gene FLOWERING LOCUS T (FT) that several flowering pathways converge to is directly regulated by more than ten transcription factors. However, very little is known about the transcriptional regulation of the FT homolog SINGLE FLOWER TRUESS (SFT) in the day-neutral plant tomato (Solanum lycopersicum). Previously, we showed that the zinc finger transcription factor SlZFP2 plays important roles in regulation of seed germination and fruit ripening in tomato and also found that overexpression of SlZFP2 impacted flowering and branching. Here, we characterized in detail the early flowering and high branching phenotypes by overexpression of this transcription factor. Our data showed that overexpression of SlZFP2 accelerated flowering in an SFT-dependent manner as demonstrated by elevated SFT expression in the leaves and the transcription factor's binding ability to SFT promoter in vitro and in vivo. Furthermore, overexpression of the SlZFP2 gene in the sft plants failed to rescue the mutant's late flowering. Through analysis of grafting phenotype, growth response of branches to auxin application and transcriptome profiling by RNA sequencing, we also showed that overexpression of SlZFP2 affected shoot apical dominance through multiple regulatory pathways. Our results suggest that the transcription factor SlZFP2 has potential applications in genetic modification of plant architecture and flowering time for tomato production and other crops as well. © 2016 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.

  17. A novel zinc finger protein Zfp277 mediates transcriptional repression of the Ink4a/arf locus through polycomb repressive complex 1

    DEFF Research Database (Denmark)

    Negishi, Masamitsu; Saraya, Atsunori; Mochizuki, Shinobu


    . METHODOLOGY/PRINCIPAL FINDINGS: We examined the function of Zinc finger domain-containing protein 277 (Zfp277), a novel zinc finger protein that interacts with the PcG protein Bmi1. Zfp277 binds to the Ink4a/Arf locus in a Bmi1-independent manner and interacts with polycomb repressor complex (PRC) 1 through...... is essential for the recruitment of PRC1 to the Ink4a/Arf locus. Our findings also highlight dynamic regulation of both Zfp277 and PcG proteins by the oxidative stress pathways....

  18. The use of ZFP lossy floating point data compression in tornado-resolving thunderstorm simulations (United States)

    Orf, L.


    In the field of atmospheric science, numerical models are used to produce forecasts of weather and climate and serve as virtual laboratories for scientists studying atmospheric phenomena. In both operational and research arenas, atmospheric simulations exploiting modern supercomputing hardware can produce a tremendous amount of data. During model execution, the transfer of floating point data from memory to the file system is often a significant bottleneck where I/O can dominate wallclock time. One way to reduce the I/O footprint is to compress the floating point data, which reduces amount of data saved to the file system. In this presentation we introduce LOFS, a file system developed specifically for use in three-dimensional numerical weather models that are run on massively parallel supercomputers. LOFS utilizes the core (in-memory buffered) HDF5 driver and includes compression options including ZFP, a lossy floating point data compression algorithm. ZFP offers several mechanisms for specifying the amount of lossy compression to be applied to floating point data, including the ability to specify the maximum absolute error allowed in each compressed 3D array. We explore different maximum error tolerances in a tornado-resolving supercell thunderstorm simulation for model variables including cloud and precipitation, temperature, wind velocity and vorticity magnitude. We find that average compression ratios exceeding 20:1 in scientifically interesting regions of the simulation domain produce visually identical results to uncompressed data in visualizations and plots. Since LOFS splits the model domain across many files, compression ratios for a given error tolerance can be compared across different locations within the model domain. We find that regions of high spatial variability (which tend to be where scientifically interesting things are occurring) show the lowest compression ratios, whereas regions of the domain with little spatial variability compress

  19. Genome-wide DNA methylation analysis of transient neonatal diabetes type 1 patients with mutations in ZFP57. (United States)

    Bak, Mads; Boonen, Susanne E; Dahl, Christina; Hahnemann, Johanne M D; Mackay, Deborah J D G; Tümer, Zeynep; Grønskov, Karen; Temple, I Karen; Guldberg, Per; Tommerup, Niels


    Transient neonatal diabetes mellitus 1 (TNDM1) is a rare imprinting disorder characterized by intrautering growth retardation and diabetes mellitus usually presenting within the first six weeks of life and resolves by the age of 18 months. However, patients have an increased risk of developing diabetes mellitus type 2 later in life. Transient neonatal diabetes mellitus 1 is caused by overexpression of the maternally imprinted genes PLAGL1 and HYMAI on chromosome 6q24. One of the mechanisms leading to overexpression of the locus is hypomethylation of the maternal allele of PLAGL1 and HYMAI. A subset of patients with maternal hypomethylation at PLAGL1 have hypomethylation at additional imprinted loci throughout the genome, including GRB10, ZIM2 (PEG3), MEST (PEG1), KCNQ1OT1 and NESPAS (GNAS-AS1). About half of the TNDM1 patients carry mutations in ZFP57, a transcription factor involved in establishment and maintenance of methylation of imprinted loci. Our objective was to investigate whether additional regions are aberrantly methylated in ZFP57 mutation carriers. Genome-wide DNA methylation analysis was performed on four individuals with homozygous or compound heterozygous ZFP57 mutations, three relatives with heterozygous ZFP57 mutations and five controls. Methylation status of selected regions showing aberrant methylation in the patients was verified using bisulfite-sequencing. We found large variability among the patients concerning the number and identity of the differentially methylated regions, but more than 60 regions were aberrantly methylated in two or more patients and a novel region within PPP1R13L was found to be hypomethylated in all the patients. The hypomethylated regions in common between the patients are enriched for the ZFP57 DNA binding motif. We have expanded the epimutational spectrum of TNDM1 associated with ZFP57 mutations and found one novel region within PPP1R13L which is hypomethylated in all TNDM1 patients included in this study. Functional

  20. Gambogic Acid Efficiently Kills Stem-Like Colorectal Cancer Cells by Upregulating ZFP36 Expression

    Directory of Open Access Journals (Sweden)

    Fang Wei


    Full Text Available Background/Aims: Gambogic acid (GA, the main active compound of Gamboge hanburyi, has been reported to be a potential novel antitumor drug. Whether GA inhibits putative cancer stem cells (CSCs, which are considered to be the major cause of cancer treatment failure, remains largely unknown. This study investigated whether GA inhibits the CSCs of colorectal cancer (CRC and its possible mechanisms. Methods: We performed CCK8 and tumor sphere formation assays, percentage analysis of both side population and CD133+CD44+ cells, and the detection of stem cells markers, in order to assess the role of GA in inhibiting the stem celllike features of CRC. An mRNA microarray was performed to identify the downstream gene affected by GA and rescue assays were performed to further clarify whether the downstream gene is involved in the GA induced decrease of the stem cell-like CRC population. CRC cells were engineered with a CSC detector vector encoding GFP and luciferase (Luc under the control of the Nanog promoter, which were utilized to investigate the effect of GA on putative CSC in human tumor xenograft-bearing mice using in vivo bioluminescence imaging. Results: Our results showed that GA significantly reduced tumor sphere formation and the percentages of side population and CD133+CD44+ cells, while also decreasing the expression of stemness and EMT-associated markers in CRC cells in vitro. GA killed stem-like CRC cells by upregulating the expression of ZFP36, which is dependent on the inactivation of the EGFR/ ERK signaling pathway. GFP+ cells harboring the PNanog-GFP-T2A-Luc transgene exhibited CSC characteristics. The in vivo results showed that GA significantly inhibited tumor growth in nude mice, accompanied by a remarkable reduction in the putative CSC number, based on whole-body bioluminescence imaging. Conclusion: These findings suggest that GA significantly inhibits putative CSCs of CRC both in vitro and in vivo by inhibiting the activation of the

  1. GhZFP1, a novel CCCH-type zinc finger protein from cotton, enhances salt stress tolerance and fungal disease resistance in transgenic tobacco by interacting with GZIRD21A and GZIPR5. (United States)

    Guo, Ying-Hui; Yu, Yue-Ping; Wang, Dong; Wu, Chang-Ai; Yang, Guo-Dong; Huang, Jin-Guang; Zheng, Cheng-Chao


    * Zinc finger proteins are a superfamily involved in many aspects of plant growth and development. However, CCCH-type zinc finger proteins involved in plant stress tolerance are poorly understood. * A cDNA clone designated Gossypium hirsutum zinc finger protein 1 (GhZFP1), which encodes a novel CCCH-type zinc finger protein, was isolated from a salt-induced cotton (G. hirsutum) cDNA library using differential hybridization screening and further studied in transgenic tobacco Nicotiana tabacum cv. NC89. Using yeast two-hybrid screening (Y2H), proteins GZIRD21A (GhZFP1 interacting and responsive to dehydration protein 21A) and GZIPR5 (GhZFP1 interacting and pathogenesis-related protein 5), which interacted with GhZFP1, were isolated. * GhZFP1 contains two typical zinc finger motifs (Cx8Cx5Cx3H and Cx5Cx4Cx3H), a putative nuclear export sequence (NES) and a potential nuclear localization signal (NLS). Transient expression analysis using a GhZFP1::GFP fusion gene in onion epidermal cells indicated a nuclear localization for GhZFP1. RNA blot analysis showed that the GhZFP1 transcript was induced by salt (NaCl), drought and salicylic acid (SA). The regions in GhZFP1 that interact with GZIRD21A and GZIPR5 were identified using truncation mutations. * Overexpression of GhZFP1 in transgenic tobacco enhanced tolerance to salt stress and resistance to Rhizoctonia solani. Therefore, it appears that GhZFP1 might be involved as an important regulator in plant responses to abiotic and biotic stresses.

  2. C3HC4-type RING finger protein NbZFP1 is involved in growth and fruit development in Nicotiana benthamiana.

    Directory of Open Access Journals (Sweden)

    Wenxian Wu

    Full Text Available C3HC4-type RING finger proteins constitute a large family in the plant kingdom and play important roles in various physiological processes of plant life. In this study, a C3HC4-type zinc finger gene was isolated from Nicotiana benthamiana. Sequence analysis indicated that the gene encodes a 24-kDa protein with 191 amino acids containing one typical C3HC4-type zinc finger domain; this gene was named NbZFP1. Transient expression of pGDG-NbZFP1 demonstrated that NbZFP1 was localized to the chloroplast, especially in the chloroplasts of cells surrounding leaf stomata. Virus-induced gene silencing (VIGS analysis indicated that silencing of NbZFP1 hampered fruit development, although the height of the plants was normal. An overexpression construct was then designed and transferred into Nicotiana benthamiana, and PCR and Southern blot showed that the NbZFP1 gene was successfully integrated into the Nicotiana benthamiana genome. The transgenic lines showed typical compactness, with a short internode length and sturdy stems. This is the first report describing the function of a C3HC4-type RING finger protein in tobacco.

  3. The ascorbate peroxidase APX1 is a direct target of a zinc finger transcription factor ZFP36 and a late embryogenesis abundant protein OsLEA5 interacts with ZFP36 to co-regulate OsAPX1 in seed germination in rice. (United States)

    Huang, Liping; Jia, Jing; Zhao, Xixi; Zhang, MengYao; Huang, Xingxiu; E Ji; Ni, Lan; Jiang, Mingyi


    Seed germination is a vital developmental process. Abscisic acid (ABA) is an essential repressor of seed germination, while ROS (reactive oxygen species) also plays a vital role in regulating seed germination. ABA could inhibit the production of ROS in seed germination, but the mechanism of ABA reduced ROS production in seed germination was hitherto unknown. Here, by ChIP (chromatin immunoprecipitation)-seq, we found that ZFP36, a rice zinc finger transcription factor, could directly bind to the promoter of OsAPX1, coding an ascorbate peroxidase (APX) which has the most affinity for H 2 O 2 (substrate; a type of ROS), and act as a transcriptional activator of OsAPX1 promoter. Moreover, ZFP36 could interact with a late embryogenesis abundant protein OsLEA5 to co-regulate the promoter activity of OsAPX1. The seed germination is highly inhibited in ZFP36 overexpression plants under ABA treatment, while an RNA interference (RNAi) mutant of OsLEA5 rice seeds were less sensitive to ABA, and exogenous ASC (ascorbate acid) could alleviate the inhibition induced by ABA. Thus, our conclusion is that OsAPX1 is a direct target of ZFP36 and OsLEA5 could interact with ZFP36 to co-regulate ABA-inhibited seed germination by controlling the expression of OsAPX1. Copyright © 2017. Published by Elsevier Inc.

  4. Genome-wide DNA methylation analysis of transient neonatal diabetes type 1 patients with mutations in ZFP57

    DEFF Research Database (Denmark)

    Bak, Mads; Boonen, Susanne E; Dahl, Christina


    of developing diabetes mellitus type 2 later in life. Transient neonatal diabetes mellitus 1 is caused by overexpression of the maternally imprinted genes PLAGL1 and HYMAI on chromosome 6q24. One of the mechanisms leading to overexpression of the locus is hypomethylation of the maternal allele of PLAGL1......BACKGROUND: Transient neonatal diabetes mellitus 1 (TNDM1) is a rare imprinting disorder characterized by intrautering growth retardation and diabetes mellitus usually presenting within the first six weeks of life and resolves by the age of 18 months. However, patients have an increased risk...... and HYMAI. A subset of patients with maternal hypomethylation at PLAGL1 have hypomethylation at additional imprinted loci throughout the genome, including GRB10, ZIM2 (PEG3), MEST (PEG1), KCNQ1OT1 and NESPAS (GNAS-AS1). About half of the TNDM1 patients carry mutations in ZFP57, a transcription factor...

  5. History and practice of material research on the examples of Material Testing and Materialpruefungsanstalt (MPA) Stuttgart, liquid crystals and screen technology as well as superconductivity. An interdisciplinary teaching project of the University of Stuttgart; Geschichte und Praxis der Materialforschung an den Beispielen Materialpruefung und Materialpruefungsanstalt (MPA) Stuttgart, Fluessigkristalle und Bildschirmtechnik sowie Supraleitung. Ein interdisziplinaeres Lehrprojekt der Universitaet Stuttgart

    Energy Technology Data Exchange (ETDEWEB)

    Hentschel, Klaus; Webel, Josef (eds.)


    The knowledge of material research and its history is not very common among scientists and engineers alike. Within the scope of an interdisciplinary teaching project carried out for the first time in the summer semester 2014 and ever since then every summer semester at the University of Stuttgart, an attempt is made to approach material research both from a scientific, technical and historical perspective. The Material Testing and Materials Testing Institute in Stuttgart (MPA), the liquid crystals and the screen technology as well as the superconductivity were selected as topics, which have a long tradition in research and teaching in Stuttgart. In this anthology the materials of the teaching project are summarized. [German] Das Wissen um die Materialforschung und ihre Geschichte ist selbst unter Naturwissenschaftlern und Ingenieuren wenig verbreitet. Im Rahmen eines erstmals im Sommersemester 2014 und seither dann jedes Sommersemester an der Universitaet Stuttgart durchgefuehrten interdisziplinaeren Lehrprojektes wird deshalb der Versuch unternommen, sich der Materialforschung sowohl aus naturwissenschaftlicher und technischer als auch aus historischer Perspektive anzunaehern. Als Themenbereiche wurden dafuer die Materialpruefung und Materialpruefungsanstalt (MPA) Stuttgart, die Fluessigkristalle und die Bildschirmtechnik sowie die Supraleitung ausgewaehlt, die in Stuttgart auf eine lange Tradition in Forschung und Lehre zurueckblicken. Im vorliegenden Sammelband sind die Materialien des Lehrprojektes zusammengefasst.

  6. Regulation of trichome development in tobacco by JcZFP8, a C2H2 zinc finger protein gene from Jatropha curcas L. (United States)

    Shi, Xiaodong; Gu, Yuxi; Dai, Tingwei; Wu, Yang; Wu, Peng; Xu, Ying; Chen, Fang


    Trichomes are epidermal outgrowths of plant tissues that can secrete or store large quantities of secondary metabolites, which contribute to plant defense responses against stress. The use of bioengineering methods for regulating the development of trichomes and metabolism is a widely researched topic. In the present study, we demonstrate that JcZFP8, a C2H2 zinc finger protein gene from Jatropha curcas L., can regulate trichome development in transgenic tobacco. To understand the underlying mechanisms, we performed transcriptome profiling of overexpression JcZFP8 transgenic plants and wild-type tobacco. Based on the analysis of differentially expressed genes, we determined that genes of the plant hormone signal transduction pathway was significantly enriched, suggesting that these pathways were modulated in the transgenic plants. In addition, the transcript levels of the known trichome-related genes in Arabidopsis were not significantly changed, whereas CycB2 and MYB genes were differentially expressed in the transgenic plants. Despite tobacco and Arabidopsis have different types of trichomes, all the pathways were associated with C2H2 zinc finger protein genes. Our findings help us to understand the regulation of multicellular trichome formation and suggest a new metabolic engineering method for the improvement of plants. Copyright © 2018 Elsevier B.V. All rights reserved.

  7. Comparative analysis of chimeric ZFP-, TALE- and Cas9-piggyBac transposases for integration into a single locus in human cells. (United States)

    Luo, Wentian; Galvan, Daniel L; Woodard, Lauren E; Dorset, Dan; Levy, Shawn; Wilson, Matthew H


    Integrating DNA delivery systems hold promise for many applications including treatment of diseases; however, targeted integration is needed for improved safety. The piggyBac (PB) transposon system is a highly active non-viral gene delivery system capable of integrating defined DNA segments into host chromosomes without requiring homologous recombination. We systematically compared four different engineered zinc finger proteins (ZFP), four transcription activator-like effector proteins (TALE), CRISPR associated protein 9 (SpCas9) and the catalytically inactive dSpCas9 protein fused to the amino-terminus of the transposase enzyme designed to target the hypoxanthine phosphoribosyltransferase (HPRT) gene located on human chromosome X. Chimeric transposases were evaluated for expression, transposition activity, chromatin immunoprecipitation at the target loci, and targeted knockout of the HPRT gene in human cells. One ZFP-PB and one TALE-PB chimera demonstrated notable HPRT gene targeting. In contrast, Cas9/dCas9-PB chimeras did not result in gene targeting. Instead, the HPRT locus appeared to be protected from transposon integration. Supplied separately, PB permitted highly efficient isolation of Cas9-mediated knockout of HPRT, with zero transposon integrations in HPRT by deep sequencing. In summary, these tools may allow isolation of 'targeted-only' cells, be utilized to protect a genomic locus from transposon integration, and enrich for Cas9-mutated cells. Published by Oxford University Press on behalf of Nucleic Acids Research 2017.

  8. The KRAB Zinc Finger Protein Roma/Zfp157 Is a Critical Regulator of Cell-Cycle Progression and Genomic Stability

    Directory of Open Access Journals (Sweden)

    Teresa L.F. Ho


    Full Text Available Regulation of DNA replication and cell division is essential for tissue growth and maintenance of genomic integrity and is particularly important in tissues that undergo continuous regeneration such as mammary glands. We have previously shown that disruption of the KRAB-domain zinc finger protein Roma/Zfp157 results in hyperproliferation of mammary epithelial cells (MECs during pregnancy. Here, we delineate the mechanism by which Roma engenders this phenotype. Ablation of Roma in MECs leads to unscheduled proliferation, replication stress, DNA damage, and genomic instability. Furthermore, mouse embryonic fibroblasts (MEFs depleted for Roma exhibit downregulation of p21Cip1 and geminin and have accelerated replication fork velocities, which is accompanied by a high rate of mitotic errors and polyploidy. In contrast, overexpression of Roma in MECs halts cell-cycle progression, whereas siRNA-mediated p21Cip1 knockdown ameliorates, in part, this phenotype. Thus, Roma is an essential regulator of the cell cycle and is required to maintain genomic stability.

  9. Nondestructive determination of the load bearing capacity of rigid pavement structures. Zerstoerungsfreie Tragfaehigkeitsbestimmung fuer starre Deckenkonstruktionen

    Energy Technology Data Exchange (ETDEWEB)

    Heinrich, U. (Bauakademie, IHLGB, Berlin (Germany))


    Actual parameters of the bearing capacity of traffic areas are in case of intensive use an indispensable necessity for the determination of the remaining use value with the aim to optimize reinforcing layers and reconstruction periods. Until now there is only one adequate measuring method for flexible layers. With the Rollig load test (RLT) initial data can be gained for the calculation of the load bearing capacity of rigid pavement structures. The method is described with the example of an airplane runway. (BWI).

  10. Non-destructive profile measurement of intensive heavy ion beams; Zerstoerungsfreie Profilmessung intensiver Schwerionenstrahlen

    Energy Technology Data Exchange (ETDEWEB)

    Becker, Frank


    Within the framework of the FAIR-project (Facility for Antiproton and Ion Research) at GSI (Helmholtz Center for Heavy Ion Research), high intensity beams from protons to uranium ions with kinetic energies up to 30 AGeV are foreseen. Present GSI-accelerators like the UNILAC and the Heavy Ion Synchrotron (SIS-18) with a magnetic rigidity of 18 Tm will be used as injectors for the future synchrotron (SIS-100). Their beam current will be increased by up to two orders of magnitude. An accurate beam position and beam profile measurement is mandatory for a safe operation of transport sections, in particular in front of production targets (Fragment Separator (FRS)-target, anti p-production-target and Warm Dense Matter (WDM)-targets). Conventional intercepting profile monitors will not withstand the thermal stress of intensive ion beams, particularly for low energy applications or focused beams. For transverse profile determination a non-intercepting Beam Induced Fluorescence (BIF)-monitor was developed, working with residual gas. The BIF-monitor exploits fluorescence light emitted by residual gas molecules after atomic collisions with beam ions. Fluorescence-images were recorded with an image-intensified camera system, and beam profiles were obtained by projecting these images. Within the scope of this dissertation the following topics have been investigated: The photon yield, profile shape and background contribution were determined for different ion species (H{sup +}, S{sup 6+}, Ar{sup 18+}, K{sup +}, Ni{sup 9+}, Xe{sup 48+}, Ta{sup 24+}, Au{sup 65+}, U{sup 73+}), beam energies (7.7 AkeV-750 AMeV), gas pressures (10{sup -6}-3 mbar) and gas species (N{sub 2}, He, Ne, Ar, Kr, Xe). Applying an imaging spectrograph and narrowband 10 nm interference filters, the spectral response was mapped and associated with the corresponding gas transitions. Spectrally resolved beam profiles were also obtained form the spectrographic images. Major results are the light yield showing a linear dependence on the gas-pressure for constant profile width and as well as the light yield being proportional to the differential energy loss. For nitrogen, spectral investigation shows a four times higher light yield compared to rare gas species, normalized with respect to the differential energy loss. Helium is the only rare gas that shows broadened beam profiles. All other rare gases and nitrogen show reasonable beam profiles that correspond well with each other. Furthermore the dose-distribution in a cave for beam energies ≥ 100 AMeV was measured and simulated in order to develop a shielding concept that protects the camera system against radiation damage. According to simulations the neutron dose decreases by 94 % in the center of a 1 m{sup 3} concrete cube. Possible profile distortions due to effects like momentum transfer, gas dynamics and the electrical field of the ion beam are discussed. Technical improvements are presented.

  11. 2011 annual meeting of the Deutsche Gesellschaft fuer Zerstoerungsfreie Pruefung (DGZfP). NDE in research, development and application

    International Nuclear Information System (INIS)


    This CD-ROM proceedings publication contains the papers and posters of the DGZfP annual meeting 2011, at Bremen, Germany, both in full text and as PowerPoint presentations. The following issues were discussed: Radiographic inspection, ultrasonic inspection, computerized tomography, and thermography. Ten papers were recorded as separate entries in the ENERGY database. [de

  12. 2013 annual meeting of the Deutsche Gesellschaft fuer Zerstoerungsfreie Pruefung (DGZfP). NDE in research, development and application

    International Nuclear Information System (INIS)


    This CD-ROM proceedings publication contains the papers and posters of the DGZfP annual meeting 2013, at Bremen, Germany, both in full text and as PowerPoint presentations. The following issues were discussed: computerized tomography, thermography; radiographic, ultrasonic, eddy current and microwave testing. 13 papers were recorded as separate entries in the INIS database.

  13. Thermografic measurement of crack initiation and propagation at thin sheet joints; Rissentstehung thermometrisch ermitteln. Zerstoerungsfreie Bestimmung der Rissinitiierung in mechanisch gefuegten und widerstandpunktgeschweissten Verbindungen

    Energy Technology Data Exchange (ETDEWEB)

    Bathke, W.; Stahlfeld, G. [Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin (Germany). Fachgruppe V.5 - Sicherheit in der Fuegetechnik


    This contribution demonstrates how a thermometric procedure might be applied to determine crack initiation during fatigue testing of joints at steel sheets. The procedure is based on the measurement of the temperature increase which is produced by the heat at the respective joint caused by deformation energy. Such investigations are aimed at detection of crack initiation before it becomes visible at the specimen surface. Thermografic measurements at different mechanical joints and resistance welded spots are compared and various applications are suggested. (orig.) [German] In diesem Beitrag wird gezeigt, wie sich ein thermometrisches Verfahren einsetzen laesst, um die Rissentstehung waehrend der Dauerschwingpruefung von Stahlblechen zu erfassen. Vergleichend werden Messungen an Proben, die durch Stanznieten, Clinchen und Widerstandspunktschweissen gefuegt wurden, gegenuebergestellt. Hierzu wird die am Fuegepunkt waehrend der Pruefung in Waerme umgewandelte Formaenderungsenergie kontinuierlich in Form der Temperaturerhoehung gemessen. Ziel dieser Untersuchungen ist es, solche Temperaturerhoehungen zur Erkennung der Rissentstehung zu verwenden, bevor der Riss die Blechoberflaeche erreicht hat und visuell erkennbar wird. Zudem werden verschiedene Anwendungsmoeglichkeiten vorgeschlagen. (orig.)

  14. Non-destructive testing of concrete structures with the impact-echo method; Zerstoerungsfreie Pruefung von Betonbauteilen mit dem Impact-Echo-Verfahren

    Energy Technology Data Exchange (ETDEWEB)

    Algernon, Daniel; Feistkorn, Sascha; Scherrer, Michael [SVTI Schweizerischer Verein fuer technische Inspektionen, Wallisellen (Switzerland). Nuklearinspektorat


    The impact-echo method is based on the use of elastic waves. It was developed in the 1980 for the testing of concrete structures and is currently widespread. Main application areas are the component and coating thickness measurement and detection of delaminations, voids and other defects. Specifically, the method is also used to check the injection faults of clamping channels. Another application is the determination of mechanical material parameters such as the modulus of elasticity. Since the original development of the method has undergone several enhancements. The conversion of a single-point measurement method towards a area component testing, the use by the optimized measurement data acquisition and evaluation enlarged and delivered an important prerequisite for increasing the efficiency. The use of air-coupled sensors not only increases the measurement speed but also provides advantages in rough component surfaces. The imaging analysis in conjunction with signal processing algorithms simplifies the interpretation and allows statistical evaluation. [German] Das Impact-Echo-Verfahren beruht auf der Nutzung elastischer Wellen. Es wurde in den 1980er Jahren fuer die Pruefung von Stahlbetonbauteilen entwickelt und ist derzeit weit verbreitet. Haupteinsatzgebiete sind die Bauteil- und Schichtdickenmessung sowie die Detektion von Delaminationen, Hohl- und anderen Fehlstellen. Insbesondere wird das Verfahren auch zur Pruefung des Verpresszustandes von Spannkanaelen herangezogen. Eine weitere Anwendung ist die Bestimmung mechanischer Materialparameter wie dem Elastizitaetsmodul. Seit der urspruenglichen Entwicklung hat das Verfahren verschiedene Weiterentwicklungen erfahren. Die Ueberfuehrung von einem Einzelpunktmessverfahren hin zu einer flaechigen Bauteilpruefung hat die Einsatzmoeglichkeiten durch die optimierte Messdatenaufnahme und -auswertung vergroessert und eine wichtige Voraussetzung zur Erhoehung der Leistungsfaehigkeit geliefert. Der Einsatz luftgekoppelter Sensoren erhoeht nicht nur die Messgeschwindigkeit sondern liefert auch Vorteile bei rauen Bauteiloberflaechen. Die bildgebende Auswertung in Verbindung mit Signalverarbeitungsalgorithmen vereinfacht die Interpretation und ermoeglicht eine statistische Beurteilung.

  15. Licence template for mobile handling and storage of radioactive substances for the nondestructive testing of materials; Mustergenehmigung zur ortsveraenderlichen Verwendung und Lagerung radioaktiver Stoffe im Rahmen der zerstoerungsfreien Materialpruefung

    Energy Technology Data Exchange (ETDEWEB)

    Lange, A. [Niedersaechsisches Ministerium fuer Umwelt, Energie und Klimaschutz (Germany); Schumann, J. [Landesamt fuer Arbeitsschutz, Gesundheitsschutz und technische Sicherheit, Berlin (Germany); Huhn, W.


    The Technical Committee ''Radiation Protection'' (Fachausschuss ''Strahlenschutz'') and the Laender Committee ''X-ray ordinance'' (Laenderausschuss ''Roentgenverordnung'') have appointed a working group for the formulation of licence templates for the nationwide use of X-ray equipment or handling of radioactive substances. To date, the following licence templates have been adopted: - Mobile operation of X-ray equipment under technical radiography to the coarse structural analysis in material testing; - Mobile operation of a handheld X-ray fluorescence system; - Mobile operation of a flash X-ray system; - Operation of an X-ray system for teleradiology The licence template ''Mobile handling and storage of radioactive substances for the nondestructive testing of materials'' is scheduled for publication. The licence template ''Practices in external facilities and installations'' is currently being revised. The licence template ''Mobile handling and storage of radioactive substances for the nondestructive testing of materials'' is used as an example to demonstrate the legal framework and the results of the working group.

  16. Nuclear technology in materials testing and radiation protection

    International Nuclear Information System (INIS)

    Neider, R.


    A report of the 1974 activities of the laboratories for physical and measuring technical fundamentals, radiation effects and radiation protection, application of radionuclides and testing of radioactive materials of the Bundesanstalt fuer Materialpruefung (BAM) is given. (RW/LH) [de

  17. Localization and Differential Expression of the Krüppel-Associated Box Zinc Finger Proteins 1 and 54 in Early Mouse Development

    DEFF Research Database (Denmark)

    Albertsen, Maria; Teperek, Marta; Elholm, Grethe


    -fused reporter gene into zygotes demonstrated the intracellular distribution of ZFP1-green fluorescent protein (GFP) and ZFP54-GFP colocalized with a DNA marker in the two-cell embryo. The KRAB domain was essential to colocalize with DNA, and deletion of the KRAB domain in ZFP1-GFP and ZFP54-GFP localized...

  18. Non-destructive, preclinical evaluation of root canal anatomy of human teeth with flat-panel detector volume CT (FD-VCT); Zerstoerungsfreie praeklinische Evaluation der Wurzelkanalanatomie menschlicher Zaehne mittels Flaechendetektor-Volumen-CT (FD-VCT)

    Energy Technology Data Exchange (ETDEWEB)

    Heidrich, G.; Hassepass, F.; Dullin, C.; Grabbe, E. [Universitaetsklinikum Goettingen, Abt. Diagnostische Radiologie (Germany); Attin, T.; Hannig, C. [Universitaetsklinikum Goettingen, Abt. fuer Zahnerhaltung, Praeventive Zahnheilkunde und Paradontologie (Germany)


    Purpose: Successful endodontic diagnostics and therapy call for adequate depiction of the root canal anatomy with multimodal diagnostic imaging. The aim of the present study is to evaluate visualization of the endodont with flat-panel detector volume CT (FD-VCT). Materials and methods: 13 human teeth were examined with the prototype of a FD-VCT. After data acquisition and generation of volume data sets in volume rendering technology (VRT), the findings obtained were compared to conventional X-rays and cross-section preparations of the teeth. Results: The anatomical structures of the endodont such as root canals, side canals and communications between different root canals as well as dentricles could be detected precisely with FD-VCT. The length of curved root canals was also determined accurately. The spatial resolution of the system is around 140 {mu}m. Only around 73% of the main root canals detected with FD-VCT and 87% of the roots could be visualized with conventional dental X-rays. None of the side canals, shown with FD-VCT, was detectable on conventional X-rays. In all cases the enamel and dentin of the teeth could be well delineated. No differences in image quality could be discerned between stored and freshly extracted teeth, or between primary and adult teeth. (orig.)

  19. Progress report on a NDT round robin on austenitic circumferential pipe welds; Fortschrittsbericht ueber einen ZfP-Ringversuch an austenitischen Rohrleitungs-Rundschweissnaehten

    Energy Technology Data Exchange (ETDEWEB)

    Brast, G [Preussische Elektrizitaets-AG (Preussenelektra), Hannover (Germany); Maier, H J; Knoch, P; Mletzko, U [Stuttgart Univ. (Germany). Staatliche Materialpruefungsanstalt


    The objective of the project is establish on the basis of Round Robin tests the current state of efficiency of various, defined testing methods, so that required or achievable optimizations can be defined and made. The project work up to date encompasses mon-destructive examinations of 15 austenitic welds with nominal widths DN 150/200/250 and wall thicknesses from 8 to 18 mm. Except for one test piece, (elbow/elbow), the joining welds are straight pipe to elbow welds. The results of the Round Robin tests show that the NDE detection limits for the fault examined (intercrystalline stress corrosion cracking) are in the range assumed so far, i.e. from about 20 to 25% of the wall thickness to be examined. The defect detection rates of the ultrasonic test methods applied are approx. 70% and thus are about equal in achievement with comparable international Round Robin tests (PISC; ASME/PDI, ENIQ, etc.). Clearly better are the fault detection rates of radiography. Evaluation of the individual results indicates the detection limits can be improved, by 1. reducing the misalignment of edges, 2. grinding of welds, 3. avoiding sharp notches at the root, 4. producing coaxial surfaces. (orig./CB) [Deutsch] Ein Ziel des Vorhabens ist es, mit Ringversuchen den derzeitigen Stand der Leistungsfaehigkeit einzelner Pruefverfahren und -techniken zu erkennen, um moeglicherweise notwendige Optimierungen vornehmen zu koennen. Das Vorhaben umfasst bis jetzt zerstoerungsfreie Pruefungen an 15 austenitischen Naehten mit Nennweiten DN 150/200/250 und Wandstaerken zwischen 8 und 18 mm. Mit einer Ausnahme (Bogen/Bogen) handelt es sich um Verbindungen Geradrohr/Bogen. Die Ergebnisse des Ringversuches weisen darauf hin, dass die Nachweisgrenzen der ZfP fuer den vorliegenden Fehlertyp (Interkristalline Spannungsrisskorrosion) in der bisher schon angenommenen Groessenordnung von ca. 20-25% der geprueften Wanddicke liegen. Die Fehlerauffind-Raten der US-Pruefung liegen mit ca. 70% im Rahmen

  20. 2015 DACH annual meeting. NDE in research, development and application. Abstracts; DACH-Jahrestagung 2015. ZfP in Forschung, Entwicklung und Anwendung. Vortraege

    Energy Technology Data Exchange (ETDEWEB)



    The main topics of this meeting are: Computed Tomography; Air Ultrasound; Accreditation, certification, validation; CT algorithms; Ultrasound - imaging; Education; Digital X-ray detectors; Ultrasound - Simulation; Material characterization; Train; Phased Array; Aviation; Acoustic emission testing; Leak testing; Local defect resonance; Composite materials; Building construction; Standards and regulations; Condition and production monitoring; Surface method; POD; Renewable energy; Thermography. [German] Die Themenschwerpunkte auf dieser Tagung waren: Computertomographie; Luftultraschall; Akkreditierung, Zertifizierung, Validierung; CT Algorithmen; Ultraschall - Bildgebung; Ausbildung; Digitale Roentgendetektoren; Ultraschall - Simulation; Materialcharakterisierung; Bahn; Phased Array; Luftfahrt; Schallemissionspruefung; Dichtheitspruefung; Lokale Defektresonanz; Verbundwerkstoffe; Bauwesen; Normen und Regelwerke; Zustands- und Fertigungsueberwachung; Oberflaechenverfahren; POD; Erneuerbare Energien; Thermographie.

  1. Effect of the linkers between the zinc fingers in zinc finger protein 809 on gene silencing and nuclear localization

    Energy Technology Data Exchange (ETDEWEB)

    Ichida, Yu, E-mail:; Utsunomiya, Yuko; Onodera, Masafumi


    Zinc finger protein 809 (ZFP809) belongs to the Kruppel-associated box-containing zinc finger protein (KRAB-ZFP) family and functions in repressing the expression of Moloney murine leukemia virus (MoMLV). ZFP809 binds to the primer-binding site (PBS)located downstream of the MoMLV-long terminal repeat (LTR) and induces epigenetic modifications at integration sites, such as repressive histone modifications and de novo DNA methylation. KRAB-ZFPs contain consensus TGEKP linkers between C2H2 zinc fingers. The phosphorylation of threonine residues within linkers leads to the inactivation of zinc finger binding to target sequences. ZFP809 also contains consensus linkers between zinc fingers. However, the function of ZFP809 linkers remains unknown. In the present study, we constructed ZFP809 proteins containing mutated linkers and examined their ability to silence transgene expression driven by MLV, binding ability to MLV PBS, and cellular localization. The results of the present study revealed that the linkers affected the ability of ZFP809 to silence transgene expression. Furthermore, this effect could be partly attributed to changes in the localization of ZFP809 proteins containing mutated linkers. Further characterization of ZFP809 linkers is required for understanding the functions and features of KRAB-ZFP-containing linkers. - Highlights: • ZFP809 has three consensus linkers between the zinc fingers. • Linkers are required for ZFP809 to silence transgene expression driven by MLV-LTR. • Linkers affect the precise nuclear localization of ZFP809.

  2. The zinc finger transcription factor 191 is required for early embryonic development and cell proliferation

    International Nuclear Information System (INIS)

    Li Jianzhong; Chen Xia; Yang Hua; Wang Shuiliang; Guo Baoyu; Yu Long; Wang Zhugang; Fu Jiliang


    Human zinc finger protein 191 (ZNF191/ZNF24) was cloned and characterized as a SCAN family member, which shows 94% identity to its mouse homologue zinc finger protein 191 (Zfp191). ZNF191 can specifically interact with an intronic polymorphic TCAT repeat (HUMTH01) in the tyrosine hydroxylase (TH) gene. Allelic variations of HUMTH01 have been stated to have a quantitative silencing effect on TH gene expression and to correlate with quantitative and qualitative changes in the binding by ZNF191. Zfp191 is widely expressed during embryonic development and in multiple tissues and organs in adult. To investigate the functions of Zfp191 in vivo, we have used homologous recombination to generate mice that are deficient in Zfp191. Heterozygous Zfp191 +/- mice are normal and fertile. Homozygous Zfp191 -/- embryos are severely retarded in development and die at approximately 7.5 days post-fertilization. Unexpectedly, in Zfp191 -/- and Zfp191 +/- embryos, TH gene expression is not affected. Blastocyst outgrowth experiments and the RNA interference-mediated knockdown of ZNF191 in cultured cells revealed an essential role for Zfp191 in cell proliferation. In further agreement with this function, no viable Zfp191 -/- cell lines were obtained by derivation of embryonic stem (ES) cells from blastocysts of Zfp191 +/- intercrosses or by forced homogenotization of heterozygous ES cells at high concentrations of G418. These data show that Zfp191 is indispensable for early embryonic development and cell proliferation

  3. An A20/AN1-type zinc finger protein modulates gibberellins and abscisic acid contents and increases sensitivity to abiotic stress in rice (Oryza sativa). (United States)

    Zhang, Ye; Lan, Hongxia; Shao, Qiaolin; Wang, Ruqin; Chen, Hui; Tang, Haijuan; Zhang, Hongsheng; Huang, Ji


    The plant hormones gibberellins (GA) and abscisic acid (ABA) play important roles in plant development and stress responses. Here we report a novel A20/AN1-type zinc finger protein ZFP185 involved in GA and ABA signaling in the regulation of growth and stress response. ZFP185 was constitutively expressed in various rice tissues. Overexpression of ZFP185 in rice results in a semi-dwarfism phenotype, reduced cell size, and the decrease of endogenous GA3 content. By contrast, higher GA3 content was observed in RNAi plants. The application of exogenous GA3 can fully rescue the semi-dwarfism phenotype of ZFP185 overexpressing plants, suggesting the negative role of ZFP185 in GA biosynthesis. Besides GA, overexpression of ZFP185 decreased ABA content and expression of several ABA biosynthesis-related genes. Moreover, it was found that ZFP185, unlike previously known A20/AN1-type zinc finger genes, increases sensitivity to drought, cold, and salt stresses, implying the negative role of ZFP185 in stress tolerance. ZFP185 was localized in the cytoplasm and lacked transcriptional activation potential. Our study suggests that ZFP185 regulates plant growth and stress responses by affecting GA and ABA biosynthesis in rice. © The Author 2015. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For permissions, please email:

  4. Non-destructive detection of fatigue in austenitic steel X6CrNiNb1810 in the HCF and VHCF range. Final report; Zerstoerungsfreie Detektion der Ermuedung von austenitischem Stahl X6CrNiNb1810 im HCF- und VHCF-Bereich. Abschlussbericht

    Energy Technology Data Exchange (ETDEWEB)

    Seiler, Georg; Boller, Christian [Univ. des Saarlandes, Saarbruecken (Germany). Lehrstuhl fuer Zerstoerungsfreie Pruefung und Qualitaetsmanagement


    In addition to load cycles in the Low Cycle Fatigue (LCF)-regime due to start up and shut down procedures of power plants, in some components additional high-frequency loadings in the High Cycle Fatigue (HCF)- and Very High Cycle Fatigue (VHCF)-regime occur. These loadings are induced e.g. by stresses due to thermal cyclic fluctuations and fluid dynamic processes. Therefore it is necessary to characterize experimentally the cyclic deformation behavior of metastable austenitic steels at operating temperature particularly in the HCF- and VHCF-regime and to develop a nondestructive method to detect fatigue processes. This joint research project was conducted in cooperation between the Institute of Materials Science and Engineering (WKK) of the University of Kaiserslautern and the Chair of Non-Destructive Testing and Quality Assurance of Saarland University (LZfPQ). WKK was focused on experimental investigations of the cyclic deformation behavior of metastable austenitic steel in the HCF- and VHCF-range, taking into account cyclic hardening and softening processes and in particular to consider fatigue-induced changes in microstructure. The samples were taken from a real pressurized water reactor component (surgeline). LZfPQ has focused on the development and application of a testing concept based on electromagnetic ultrasonic measurements. Within this subproject, electromagnetic acoustic transducers (EMATs) for in situ Operation in WKK's fatigue testing equipment have been developed and assembled. Single-step fatigue tests at elevated temperature (300 C) were conducted both with total-stress and strain control. During the test, the fatigue specimens were characterized by EMAT ultrasound in transmission mode. Different sensor types and wavemodes were used. The transmission signal was evaluated in terms of time of flight and amplitude. Both measured values are seen to behave in a characteristic manner depending on the test duration and the mechanical measured value (stress or strain amplitude, respectively) and thus also depending on the fatigue state. In particular, early prediction of the specimen failure is possible on this basis. The EMAT approach is thus a potential method for non-destructive early fatigue detection within an aging management system for power plants and should be further pursued.

  5. Marketingová strategie ve vybrané organizaci


    Krejčová, Kamila


    The main objective of my bachelor's thesis "Marketing strategy in the company ZFP akademie, a. s." is the analyses of marketing strategy of the company ZFP akademie, a.s. and its product portfolio. Than I make its marketing strategy clear and show the future possible development.

  6. Mouse Homologue of the Schizophrenia Susceptibility Gene ZNF804A as a Target of Hoxc8

    Directory of Open Access Journals (Sweden)

    Hyun Joo Chung


    Full Text Available Using a ChIP-cloning technique, we identified a Zinc finger protein 804a (Zfp804a as one of the putative Hoxc8 downstream target genes. We confirmed binding of Hoxc8 to an intronic region of Zfp804a by ChIP-PCR in F9 cells as well as in mouse embryos. Hoxc8 upregulated Zfp804a mRNA levels and augmented minimal promoter activity in vitro. In E11.5 mouse embryos, Zfp804a and Hoxc8 were coexpressed. Recent genome-wide studies identified Zfp804a (or ZNF804A in humans as a plausible marker for schizophrenia, leading us to hypothesize that this embryogenic regulatory control might also exert influence in development of complex traits such as psychosis.

  7. The Rat Homolog of the Schizophrenia Susceptibility Gene ZNF804A Is Highly Expressed during Brain Development, Particularly in Growth Cones

    DEFF Research Database (Denmark)

    Hinna, Katja Hvid; Rich, Karen; Fex Svenningsen, Åsa


    it decreases towards adult levels. This time point is developmentally the equivalent to the second trimester of fetal development in humans. An exception to this expression pattern is the hippocampus where the expression of Zfp804A appears to increase again in the adult brain. Using laser capture...... developmental mechanisms are suggested in the pathophysiology for schizophrenia, expression of Zfp804A, the rat homolog of ZNF804A, was investigated in the developing rat brain. We found that expression of Zfp804A in most brain regions is developmentally regulated and peaks around birth, where after...... expression was therefore investigated with immunochemistry in such cultures. Interestingly, before day 4, the protein is mostly found in the perinuclear region of the cell but at day 4, ZFP804A was instead found throughout the cell and particularly in the growth cones. In conclusion we demonstrate that Zfp...

  8. 32. MPA seminar 2006. Lectures; 32. MPA Seminar 2006. Vortraege

    Energy Technology Data Exchange (ETDEWEB)



    This conference CD contains the full text of 47 papers on the following subjects: 1. Ageing management; 2. Integrity analysis; 3. Materials engineering; 4. State monitoring; 5. Pressure equipment status monitoring; 6. Quality assurance; 7. Non-destructive testing; 8. Fracture-mechanical analysis of components. [German] Diese Tagungs-CD enthaelt den Text von 47 Vortraegen mit folgenden Themenschwerpunkten: 1. Alterungsmanagement; 2. Integritaetsanalyse; 3. Werkstofftechnik; 4. Zustandsueberwachung; 5. Beschaffenheit Druckgeraete; 6. Qualitaetssicherung; 7. Zerstoerungsfreie Pruefung; und 8. Bruchmechanische Bauteilanalyse.

  9. Some aspects of industrial non-destructive evaluation by X- and γ-ray computed tomography

    International Nuclear Information System (INIS)

    Reimers, P.; Goebbels, J.; Weise, H.P.; Wilding, K.


    The development of an industrial CAT-Scanner at the Bundesanstalt fuer Materialpruefung (BAM) is described and some practical results on typical nondestructive testing problems are presented. General criteria for the image quality, especially spatial and density resolution, are discussed on the basis of the appropriate mathematical relationships. The limiting parameter in industrial CAT will be the final specific photon emissivity of available radiation sources. Therefore the relative density resolution obtainable with the three most common types of radiation sources is calculated by variation of material thickness and scanning time. (orig.)

  10. The Rat Homolog of the Schizophrenia Susceptibility Gene ZNF804A Is Highly Expressed during Brain Development, Particularly in Growth Cones.

    Directory of Open Access Journals (Sweden)

    Katja Hvid Hinna

    Full Text Available A single nucleotide polymorphism in the ZNF804A gene, rs1344706, is associated with schizophrenia. The polymorphism has been suggested to alter fetal expression of ZNF804A. It has also been reported to be associated with altered cortical functioning and neural connectivity in the brain. Since developmental mechanisms are suggested in the pathophysiology for schizophrenia, expression of Zfp804A, the rat homolog of ZNF804A, was investigated in the developing rat brain. We found that expression of Zfp804A in most brain regions is developmentally regulated and peaks around birth, where after it decreases towards adult levels. This time point is developmentally the equivalent to the second trimester of fetal development in humans. An exception to this expression pattern is the hippocampus where the expression of Zfp804A appears to increase again in the adult brain. Using laser capture and quantitative PCR we found that Zfp804A mRNA expression in the adult rat hippocampus is highest in the CA1 sub region, where the overall firing rates of neurons is higher than in the CA3 region. In cultured cortical neurons Zfp804A mRNA expression peaked at day 4 and then decreased. The ZFP804A protein expression was therefore investigated with immunochemistry in such cultures. Interestingly, before day 4, the protein is mostly found in the perinuclear region of the cell but at day 4, ZFP804A was instead found throughout the cell and particularly in the growth cones. In conclusion we demonstrate that Zfp804A increases in the rat brain at the time of birth, coinciding with neuronal differentiation. We also show that ZFP804A is localized to growth cones of growing neurites. These data implicate ZFP804A in growth cone function and neurite elongation. The polymorphism rs1344706 lowers expression of ZNF804A during prenatal brain development. This may affect ZNF804A's role in cone function and neurite elongation leading to synaptic deficits and altered neural connectivity.

  11. Green tea increases anti-inflammatory tristetraprolin and decreases pro-inflammatory tumor necrosis factor mRNA levels in rats

    Directory of Open Access Journals (Sweden)

    Roussel Anne M


    Full Text Available Abstract Background Tristetraprolin (TTP/ZFP36 family proteins have anti-inflammatory activity by binding to and destabilizing pro-inflammatory mRNAs such as Tnf mRNA, and represent a potential therapeutic target for inflammation-related diseases. Tea has anti-inflammatory properties but the molecular mechanisms have not been completely elucidated. We hypothesized that TTP and/or its homologues might contribute to the beneficial effects of tea as an anti-inflammatory product. Methods Quantitative real-time PCR was used to investigate the effects of green tea (0, 1, and 2 g solid extract/kg diet on the expression of Ttp family genes (Ttp/Tis11/Zfp36, Zfp36l1/Tis11b, Zfp36l2/Tis11d, Zfp36l3, pro-inflammatory genes (Tnf, Csf2/Gm-csf, Ptgs2/Cox2, and Elavl1/Hua/Hur and Vegf genes in liver and muscle of rats fed a high-fructose diet known to induce insulin resistance, oxidative stress, inflammation, and TNF-alpha levels. Results Ttp and Zfp36l1 mRNAs were the major forms in both liver and skeletal muscle. Ttp, Zfp36l1, and Zfp36l2 mRNA levels were more abundant in the liver than those in the muscle. Csf2/Gm-csf and Zfp36l3 mRNAs were undetectable in both tissues. Tea (1 g solid extract/kg diet increased Ttp mRNA levels by 50–140% but Tnf mRNA levels decreased by 30% in both tissues, and Ptgs2/Cox2 mRNA levels decreased by 40% in the muscle. Tea (2 g solid extract/kg diet increased Elavl1/Hua/Hur mRNA levels by 40% in the liver but did not affect any of the other mRNA levels in liver or muscle. Conclusion These results show that tea can modulate Ttp mRNA levels in animals and suggest that a post-transcriptional mechanism through TTP could partially account for tea's anti-inflammatory properties. The results also suggest that drinking adequate amounts of green tea may play a role in the prevention of inflammation-related diseases.

  12. Bundesanstalt fuer Martialforschung und -pruefung (BAM). Annual report 2001; Bundesanstalt fuer Materialforschung und -pruefung (BAM). Jahresbericht 2001

    Energy Technology Data Exchange (ETDEWEB)



    The volume reports the activities of the BAM, (Federal Office for Materials Research and Testing), and its departments for Analytical Chemistry/Reference Materials, Chemical Safety Engineering, Hazardous Materials Containment, Environmental Compatibility of Materials, Technology of Structural Materials, Polymers, Structural Stability and Safety, Materials Protection/Nondestructive Testing, and for Scientific-Technical Interdisciplinary Activities. (MM) [German] Der vorliegende Band berichtet ueber die Aktivitaeten der Bundesanstalt fuer Materialforschung und -pruefung und ihrer Abteilungen Analytische Chemie/Referenzmaterialien, Chemische Sicherheitstechnik, Gefahrgutumschliessungen, Umweltvertraeglichkeit von Materialien, Werkstofftechnik der Konstruktionswerkstoffe, Funktion von Polymeren, Bauwerkssicherheit, Materialschutz/Zerstoerungsfreie Pruefung und Technisch-wissenschaftliche Querschnittsfunktionen. (MM)

  13. A conserved PHD finger protein and endogenous RNAi modulate insulin signaling in Caenorhabditis elegans. (United States)

    Mansisidor, Andres R; Cecere, Germano; Hoersch, Sebastian; Jensen, Morten B; Kawli, Trupti; Kennedy, Lisa M; Chavez, Violeta; Tan, Man-Wah; Lieb, Jason D; Grishok, Alla


    Insulin signaling has a profound effect on longevity and the oxidative stress resistance of animals. Inhibition of insulin signaling results in the activation of DAF-16/FOXO and SKN-1/Nrf transcription factors and increased animal fitness. By studying the biological functions of the endogenous RNA interference factor RDE-4 and conserved PHD zinc finger protein ZFP-1 (AF10), which regulate overlapping sets of genes in Caenorhabditis elegans, we identified an important role for these factors in the negative modulation of transcription of the insulin/PI3 signaling-dependent kinase PDK-1. Consistently, increased expression of pdk-1 in zfp-1 and rde-4 mutants contributed to their reduced lifespan and sensitivity to oxidative stress and pathogens due to the reduction in the expression of DAF-16 and SKN-1 targets. We found that the function of ZFP-1 in modulating pdk-1 transcription was important for the extended lifespan of the age-1(hx546) reduction-of-function PI3 kinase mutant, since the lifespan of the age-1; zfp-1 double mutant strain was significantly shorter compared to age-1(hx546). We further demonstrate that overexpression of ZFP-1 caused an increased resistance to oxidative stress in a DAF-16-dependent manner. Our findings suggest that epigenetic regulation of key upstream signaling components in signal transduction pathways through chromatin and RNAi may have a large impact on the outcome of signaling and expression of numerous downstream genes.

  14. Differential trypanosome surface coat regulation by a CCCH protein that co-associates with procyclin mRNA cis-elements.

    Directory of Open Access Journals (Sweden)

    Pegine Walrad


    Full Text Available The genome of Trypanosoma brucei is unusual in being regulated almost entirely at the post-transcriptional level. In terms of regulation, the best-studied genes are procyclins, which encode a family of major surface GPI-anchored glycoproteins (EP1, EP2, EP3, GPEET that show differential expression in the parasite's tsetse-fly vector. Although procyclin mRNA cis-regulatory sequences have provided the paradigm for post-transcriptional control in kinetoplastid parasites, trans-acting regulators of procyclin mRNAs are unidentified, despite intensive effort over 15 years. Here we identify the developmental regulator, TbZFP3, a CCCH-class predicted RNA binding protein, as an isoform-specific regulator of Procyclin surface coat expression in trypanosomes. We demonstrate (i that endogenous TbZFP3 shows sequence-specific co-precipitation of EP1 and GPEET, but not EP2 and EP3, procyclin mRNA isoforms, (ii that ectopic overexpression of TbZFP3 does not perturb the mRNA abundance of procyclin transcripts, but rather that (iii their protein expression is regulated in an isoform-specific manner, as evidenced by mass spectrometric analysis of the Procyclin expression signature in the transgenic cell lines. The TbZFP3 mRNA-protein complex (TbZFP3mRNP is identified as a trans-regulator of differential surface protein expression in trypanosomes. Moreover, its sequence-specific interactions with procyclin mRNAs are compatible with long-established predictions for Procyclin regulation. Combined with the known association of TbZFP3 with the translational apparatus, this study provides a long-sought missing link between surface protein cis-regulatory signals and the gene expression machinery in trypanosomes.

  15. Report on the joint USA-Germany drop test program for a vitrified high level waste cask

    International Nuclear Information System (INIS)

    Golliher, K.G.; Witt, C.R.; Wieser, K.E.


    A series of full-scale drop tests was performed on a ductile iron transport cask in a cooperative program between the US Department of Energy (DOE) and Bundesantalt fuer Materialpruefung (BAM) in Germany. The tests, which were performed at BAM's test facility located near Lehre, Germany, were performed on a prototype cask designed for transport of Vitrified High Level Waste (VHLW) canisters. The VHLW cask is a right circular cylinder with a diameter of 1156 mm and a height of 3454 cm, and weighs approximately 24.6 kg including its payload of a single VHLW canister. The drop tests were performed with a non-radioactive, prototype VHLW canister in the cavity. (J.P.N.)

  16. Targeting artificial transcription factors to the utrophin A promoter: effects on dystrophic pathology and muscle function. (United States)

    Lu, Yifan; Tian, Chai; Danialou, Gawiyou; Gilbert, Rénald; Petrof, Basil J; Karpati, George; Nalbantoglu, Josephine


    Duchenne muscular dystrophy is caused by a genetic defect in the dystrophin gene. The absence of dystrophin results in muscle fiber necrosis and regeneration, leading to progressive muscle fiber loss. Utrophin is a close analogue of dystrophin. A substantial, ectopic expression of utrophin in the extrasynaptic sarcolemma of dystrophin-deficient muscle fibers can prevent deleterious effects of dystrophin deficiency. An alternative approach for the extrasynaptic up-regulation of utrophin involves the augmentation of utrophin transcription via the endogenous utrophin A promoter using custom-designed transcriptional activator proteins with zinc finger (ZFP) motifs. We tested a panel of custom-designed ZFP for their ability to activate the utrophin A promoter. Expression of one such ZFP efficiently increased, in a time-dependent manner, utrophin transcript and protein levels both in vitro and in vivo. In dystrophic mouse (mdx) muscles, administration of adenoviral vectors expressing this ZFP led to significant enhancement of muscle function with decreased necrosis, restoration of the dystrophin-associated proteins, and improved resistance to eccentric contractions. These studies provide evidence that specifically designed ZFPs can act as strong transcriptional activators of the utrophin A promoter. These may thus serve as attractive therapeutic agents for dystrophin deficiency states such as Duchenne muscular dystrophy.

  17. Presumed pluripotency markers UTF-1 and REX-1 are expressed in human adult testes and germ cell neoplasms

    DEFF Research Database (Denmark)

    Kristensen, David M; Nielsen, John E; Skakkebaek, Niels E


    UTF-1 and REX-1/ZFP42 are transcription factors involved in pluripotency. Because of phenotypic similarities between pluripotent embryonic stem cells and testicular germ cell tumours (TGCT) and the derivation of pluripotent cells from testes, we investigated the expression of UTF-1 and REX-1 during...... human gonadal development and in TGCT....

  18. Cloning and analysis of the mouse Fanconi anemia group A cDNA and an overlapping penta zinc finger cDNA. (United States)

    Wong, J C; Alon, N; Norga, K; Kruyt, F A; Youssoufian, H; Buchwald, M


    Despite the cloning of four disease-associated genes for Fanconi anemia (FA), the molecular pathogenesis of FA remains largely unknown. To study FA complementation group A using the mouse as a model system, we cloned and characterized the mouse homolog of the human FANCA cDNA. The mouse cDNA (Fanca) encodes a 161-kDa protein that shares 65% amino acid sequence identity with human FANCA. Fanca is located at the distal region of mouse chromosome 8 and has a ubiquitous pattern of expression in embryonic and adult tissues. Expression of the mouse cDNA in human FA-A cells restores the cellular drug sensitivity to normal levels. Thus, the expression pattern, protein structure, chromosomal location, and function of FANCA are conserved in the mouse. We also isolated a novel zinc finger protein, Zfp276, which has five C(2)H(2) domains. Interestingly, Zfp276 is situated in the Fanca locus, and the 3'UTR of its cDNA overlaps with the last four exons of Fanca in a tail-to-tail manner. Zfp276 is expressed in the same tissues as Fanca, but does not complement the mitomycin C (MMC)-sensitive phenotype of FA-A cells. The overlapping genomic organization between Zfp276 and Fanca may have relevance to the disease phenotype of FA. Copyright 2000 Academic Press.

  19. Journal of Biosciences | Indian Academy of Sciences

    Indian Academy of Sciences (India)

    Permanent link: Keywords. Epigenetics; genomic imprinting; heterochromatin; HP1; KRAB-ZFP. Abstract. Twenty five years ago it was proposed that conserved components of constitutive heterochromatin assemble heterochromatinlikecomplexes in euchromatin ...

  20. DNA Origami Scaffolds as Templates for Functional Tetrameric Kir3 K+ Channels. (United States)

    Kurokawa, Tatsuki; Kiyonaka, Shigeki; Nakata, Eiji; Endo, Masayuki; Koyama, Shohei; Mori, Emiko; Tran, Nam Ha; Dinh, Huyen; Suzuki, Yuki; Hidaka, Kumi; Kawata, Masaaki; Sato, Chikara; Sugiyama, Hiroshi; Morii, Takashi; Mori, Yasuo


    In native systems, scaffolding proteins play important roles in assembling proteins into complexes to transduce signals. This concept is yet to be applied to the assembly of functional transmembrane protein complexes in artificial systems. To address this issue, DNA origami has the potential to serve as scaffolds that arrange proteins at specific positions in complexes. Herein, we report that Kir3 K + channel proteins are assembled through zinc-finger protein (ZFP)-adaptors at specific locations on DNA origami scaffolds. Specific binding of the ZFP-fused Kir3 channels and ZFP-based adaptors on DNA origami were confirmed by atomic force microscopy and gel electrophoresis. Furthermore, the DNA origami with ZFP binding sites nearly tripled the K + channel current activity elicited by heterotetrameric Kir3 channels in HEK293T cells. Thus, our method provides a useful template to control the oligomerization states of membrane protein complexes in vitro and in living cells. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Conversion of Human Fibroblasts to Stably Self-Renewing Neural Stem Cells with a Single Zinc-Finger Transcription Factor

    Directory of Open Access Journals (Sweden)

    Ebrahim Shahbazi


    Full Text Available Direct conversion of somatic cells into neural stem cells (NSCs by defined factors holds great promise for mechanistic studies, drug screening, and potential cell therapies for different neurodegenerative diseases. Here, we report that a single zinc-finger transcription factor, Zfp521, is sufficient for direct conversion of human fibroblasts into long-term self-renewable and multipotent NSCs. In vitro, Zfp521-induced NSCs maintained their characteristics in the absence of exogenous factor expression and exhibited morphological, molecular, developmental, and functional properties that were similar to control NSCs. In addition, the single-seeded induced NSCs were able to form NSC colonies with efficiency comparable with control NSCs and expressed NSC markers. The converted cells were capable of surviving, migrating, and attaining neural phenotypes after transplantation into neonatal mouse and adult rat brains, without forming tumors. Moreover, the Zfp521-induced NSCs predominantly expressed rostral genes. Our results suggest a facilitated approach for establishing human NSCs through Zfp521-driven conversion of fibroblasts.

  2. RNA interference and retinoblastoma-related genes are required for repression of endogenous siRNA targets in Caenorhabditis elegans. (United States)

    Grishok, Alla; Hoersch, Sebastian; Sharp, Phillip A


    In Caenorhabditis elegans, a vast number of endogenous short RNAs corresponding to thousands of genes have been discovered recently. This finding suggests that these short interfering RNAs (siRNAs) may contribute to regulation of many developmental and other signaling pathways in addition to silencing viruses and transposons. Here, we present a microarray analysis of gene expression in RNA interference (RNAi)-related mutants rde-4, zfp-1, and alg-1 and the retinoblastoma (Rb) mutant lin-35. We found that a component of Dicer complex RDE-4 and a chromatin-related zinc finger protein ZFP-1, not implicated in endogenous RNAi, regulate overlapping sets of genes. Notably, genes a) up-regulated in the rde-4 and zfp-1 mutants and b) up-regulated in the lin-35(Rb) mutant, but not the down-regulated genes are highly represented in the set of genes with corresponding endogenous siRNAs (endo-siRNAs). Our study suggests that endogenous siRNAs cooperate with chromatin factors, either C. elegans ortholog of acute lymphoblastic leukemia-1 (ALL-1)-fused gene from chromosome 10 (AF10), ZFP-1, or tumor suppressor Rb, to regulate overlapping sets of genes and predicts a large role for RNAi-based chromatin silencing in control of gene expression in C. elegans.

  3. Cerebral haemodynamic physiology during steep Trendelenburg position and CO2 pneumoperitoneum

    NARCIS (Netherlands)

    Kalmar, A. F.; Dewaele, F.; Foubert, L.; Hendrickx, J. F.; Heeremans, E. H.; Struys, M. M. R. F.; Absalom, A.

    The steep (40) Trendelenburg position optimizes surgical exposure during robotic prostatectomy. The goal of the current study was to elucidate the influence of this patient positioning on cerebral blood flow and zero flow pressure (ZFP), and to assess the validity of different methods of evaluating

  4. Economic importance of non-destructive testing

    International Nuclear Information System (INIS)

    Loebert, P.


    On May 21 to 23, 1979, the annual meeting of the Deutsche Gesellschaft fuer Zerstoerungsfreie Pruefung took place in Lindau near the Bodensee lake. About 600 experts from Germany and abroad participated in the meeting, whose general subject was 'The Economic Importance of Non-Destructive Testing'. Theoretical problems and practical investigations were discussed in a number of papers on special subjects. Apart from the 33 papers, there was also a poster show with 53 stands with texts, drawings, diagrams, and figures where the authors informed those interested on the latest state of knowledge in testing. The short papers were read in six sessions under the headings of rentability of non-destructive testing, X-ray methods, electromagnetic methods, and ultrasonic methods 1 and 2. (orig.) [de

  5. Condition based maintenance of a 20 kV-PE/XLPE-insulated cable network using the IRC-Analysis; Zustandsorientierte Instandhaltung eines polymerisolierten 20-kV-Kabelnetzes mit der IRC-Analyse. Moderne Diagnostik reduziert Stoerungsrate

    Energy Technology Data Exchange (ETDEWEB)

    Hoff, G.; Kranz, H.G. [BUGH Wuppertal (Germany). Labs. fuer Hochspannungstechnik; Beigert, M.; Petzold, F. [Seba Dynatronic Mess- und Ortungstechnik GmbH, Baunach (Germany); Kneissl, C. [Bereich Konzeption und Messtechnik, Lech Elektrizitaetswerke AG, Augsburg (Germany)


    For a preventive maintenance of a polymer insulated cable network a destruction free estimation of the status of the buried PE/XLPE-cables is needed. This contribution presents a condition based maintenance concept which is based on the IRC-Analysis. With this concept a major German utility was able to reduce the amount of failures in a part of the 20kV-cable network. The general tendency of increasing faults was broken. (orig.) [German] Die praeventive Instandhaltung in polymerisolierten Kabelnetzen setzt eine zerstoerungsfreie Zustandbestimmung von gelegten PE/VPE-isolierten Kabeln voraus. Die-Verfasser beschreiben ein zustandsorientiertes Wartungskonzept auf Basis der IRC-Analyse, mit dem es gelungen ist, in einem Teil des 20-kV-Kabelnetzes eines Energieversorgers die in der Vergangenheit stetig steigende Stoerungsrate drastisch zu reduzieren. (orig.)

  6. Predicting success of oligomerized pool engineering (OPEN for zinc finger target site sequences

    Directory of Open Access Journals (Sweden)

    Goodwin Mathew J


    Full Text Available Abstract Background Precise and efficient methods for gene targeting are critical for detailed functional analysis of genomes and regulatory networks and for potentially improving the efficacy and safety of gene therapies. Oligomerized Pool ENgineering (OPEN is a recently developed method for engineering C2H2 zinc finger proteins (ZFPs designed to bind specific DNA sequences with high affinity and specificity in vivo. Because generation of ZFPs using OPEN requires considerable effort, a computational method for identifying the sites in any given gene that are most likely to be successfully targeted by this method is desirable. Results Analysis of the base composition of experimentally validated ZFP target sites identified important constraints on the DNA sequence space that can be effectively targeted using OPEN. Using alternate encodings to represent ZFP target sites, we implemented Naïve Bayes and Support Vector Machine classifiers capable of distinguishing "active" targets, i.e., ZFP binding sites that can be targeted with a high rate of success, from those that are "inactive" or poor targets for ZFPs generated using current OPEN technologies. When evaluated using leave-one-out cross-validation on a dataset of 135 experimentally validated ZFP target sites, the best Naïve Bayes classifier, designated ZiFOpT, achieved overall accuracy of 87% and specificity+ of 90%, with an ROC AUC of 0.89. When challenged with a completely independent test set of 140 newly validated ZFP target sites, ZiFOpT performance was comparable in terms of overall accuracy (88% and specificity+ (92%, but with reduced ROC AUC (0.77. Users can rank potentially active ZFP target sites using a confidence score derived from the posterior probability returned by ZiFOpT. Conclusion ZiFOpT, a machine learning classifier trained to identify DNA sequences amenable for targeting by OPEN-generated zinc finger arrays, can guide users to target sites that are most likely to function

  7. Development of new testing methods for the numerical load analysis for the drop test of steel sheet containers for the final repository Konrad

    International Nuclear Information System (INIS)

    Protz, C.; Voelzke, H.; Zencker, U.; Hagenow, P.; Gruenewald, H.


    The qualification of steel sheet containers as intermediate-level waste container for the final repository is performed by the BAM (Bundeasmt fuer Materialpruefung) according to the BfS (Bundesamt fuer Strahlenschutz) requirements. The testing requirements include the stacking pressure tests, lifting tests, drop tests thermal tests (fire resistance) and tightness tests. Besides the verification using model or prototype tests and transferability considerations numerical safety analyses may be performed alternatively. The authors describe the internal BAM research project ConDrop aimed to develop extended testing methods for the drop test of steel sheet containers for the final repository Konrad using numerical load analyses. A finite element model was developed using The FE software LS-PrePost 3.0 and ANSYS 12.0 and the software FE-Code LS-DYNA for the simulation of the drop test (5 m height). The results were verified by experimental data from instrumented drop tests. The container preserves its integrity after the drop test, plastic deformation occurred at the bottom plate, the side walls, the cask cover and the lateral uprights.

  8. Rescue the failed half-ZFN by a sensitive mammalian cell-based luciferase reporter system.

    Directory of Open Access Journals (Sweden)

    Weifeng Zhang

    Full Text Available ZFN technology is a powerful research tool and has been used for genome editing in cells lines, animals and plants. The generation of functional ZFNs for particular targets in mammalian genome is still challenging for an average research group. The modular-assembly method is relatively fast, easy-to-practice but has a high failure rate. Some recent studies suggested that a ZFP with low binding activity might be able to form a working ZFN pair with another binding active half-ZFP. In order to unveil the potential ZFP candidates among those with low binding activities, this paper established a highly sensitive mammalian cell-based transcriptional reporter system to assess the DNA binding activities of ZFPs by inserting multiple copies of ZFN target sequence fragment (TSF of an interested gene (e. g., hPGRN or hVEGF. Our results showed that this system increased the screening sensitivity up to 50-fold and markedly amplified the differences in the binding activities between different ZFPs. We also found that the targeted chromosomal gene repair efficiency of each hPGRN or hVEGF ZFN pair was in proportion with the combination of the binding activities of the ZFL (Left zinc finger and ZFR (Right zinc finger. A hPGRN ZFR with low binding ability was able to form a biological active ZFN if combined with a hPGRN ZFL with relatively high binding ability. Lastly, site-specific genome editing by hPGRN ZFNs generated by this system was confirmed by sequencing, and the PGRN knock-out cell line showed significantly decreased cell growth compared with the control. Our system will provide a valuable tool for further optimizing the nucleases with regard to specificity and cytotoxicity.

  9. Zebrafish Model of NF1 for Structure-Function Analysis, Mechanisms of Glial Tumorigenesis, and Chemical Biology (United States)


    models. Zinc-finger nucleases (ZFNs) are chimeric fusions between a zinc-finger protein (ZFP) and the nuclease domain of FokI (Urnov et al., 2010). ZFNs...reference line. Once a target site is chosen, the user can click on the ZFN entry (QueryID; see Fig. 4B) for details about each construct (Fig. 4C...Zhong, Y. (1997). Requirement of Drosophila NF1 for activation of adenylyl cyclase by PACAP38-like neuro- peptides . Science 276, 795–798.ports 8, 1265

  10. The genomic landscape of pediatric and young adult T-lineage acute lymphoblastic leukemia | Office of Cancer Genomics (United States)

    Genetic alterations that activate NOTCH1 signaling and T cell transcription factors, coupled with inactivation of the INK4/ARF tumor suppressors, are hallmarks of T-lineage acute lymphoblastic leukemia (T-ALL), but detailed genome-wide sequencing of large T-ALL cohorts has not been carried out. Using integrated genomic analysis of 264 T-ALL cases, we identified 106 putative driver genes, half of which had not previously been described in childhood T-ALL (for example, CCND3, CTCF, MYB, SMARCA4, ZFP36L2 and MYCN).

  11. Derivation of hybrid ES cell lines from two different strains of mice

    Directory of Open Access Journals (Sweden)

    Ho-Tak Lau


    Full Text Available Parental origin-dependent expression of the imprinted genes is essential for mammalian development. Zfp57 maintains genomic imprinting in mouse embryos and ES cells. To examine the allelic expression patterns of the imprinted genes in ES cells, we obtained multiple hybrid ES clones that were directly derived from the blastocysts generated from the cross between mice on two different genetic backgrounds. The blastocyst-derived ES clones displayed largely intact DNA methylation imprint at the tested imprinted regions. These hybrid ES clones will be useful for future studies to examine the allelic expression of the imprinted genes in ES cells and their differentiated progeny.

  12. Electromagnetic and ultrasonic techniques to evaluate stress states of components; Elektromagnetische und Ultraschallverfahren zur Spannungsanalyse an Bauteilen

    Energy Technology Data Exchange (ETDEWEB)

    Schneider, E.; Kern, R.; Theiner, W.A. [Fraunhofer Inst. fuer Zerstoerungsfreie Pruefverfahren, IZFP, Saarbruecken (Germany)


    The electromagnetic and ultrasonic techniques are comparably recent NDT methods for determination of stress states of components. They are simple in application, but require pre-measurement preparation: Electromagnetic techniques need calibration, and quantitative stress analysis by ultrasonic techniques needs reference values, i.e. verified materials-specific quantities to be obtained with representative specimens. Electromagnetic and ultrasonic techniques have been developed for specific tests at defined components, and the corresponding instruments and sensors have been used in practice for several years now. The paper summarizes fundamental aspects and explains the state of the art by means of several examples. (orig./CB) [Deutsch] Elektromagnetische und Ultraschallverfahren sind vergleichsweise neue zerstoerungsfreie Verfahren zur Bestimmung von Eigenspannungen in Bauteilen. Ihre Anwendung ist einfach, setzt aber Vorarbeiten voraus: Elektromagnetische Verfahren muessen kalibriert und zur quantitativen Spannungsanalyse mittels Ultraschallverfahren muessen materialspezifische Kenngroessen an repraesentativen Materialproben ermittelt werden. Elektromagnetische und Ultraschallverfahren sind fuer konkrete Anwendungen an Bauteilen entwickelt, angepasste Geraete und Sensoren seit Jahren in der Nutzung. Der Beitrag fasst die Grundlagen zusammen und stellt den Stand der Technik anhand ausgewaehlter Anwendungen dar. (orig.)

  13. Limits of applicability in NDT inspections. Practical examples; Grenzen der Anwendbarkeit bei ZfP-Pruefungen. Beispiele aus der Praxis

    Energy Technology Data Exchange (ETDEWEB)

    Sodeikat, Christian [Ingenieurbuero Schiessl Gehlen Sodeikat GmbH, Muenchen (Germany); Lohse, Carsten [Bewehrungsnachweis und -analyse, Augustusburg (Germany)


    Property owners, developers, architects, planners and in many cases courts require further information regarding the state of existing and under construction buildings. In order to avoid concerns on the building structure and to avoid nuisance by noise and dirt and ultimately save costs, structural inspection should be carried out as non-destructive. NDT methods, however, always have application limits that can be very different depending on the methods used and test equipment. However, not every inspection task, the implementation of which customer want, can be solved with NDT methods. The following article presents practical examples of application limits of different NDT methods. [German] Eigentuemer, Bauherren, Architekten, Planer und vielfach auch Gerichte benoetigen weitergehende Informationen bzgl. des Zustands bestehender und auch im Bau befindlicher Bauwerke. Um die Bausubstanz nicht zu beeintraechtigen, Belaestigungen durch Laerm und Dreck zu vermeiden und letztlich Kosten zu sparen, sollen Bauwerksuntersuchungen moeglichst zerstoerungsfrei durchgefuehrt werden. ZfP-Verfahren weisen jedoch immer Anwendungsgrenzen auf, die je nach eingesetzten Verfahren und Pruefgeraeten sehr unterschiedlich sein koennen. Jedoch kann nicht jede Pruefaufgabe, deren Umsetzung sich Auftraggeber wuenschen, mit ZfP-Verfahren geloest werden. Der nachfolgende Beitrag stellt Praxisbeispiele fuer Anwendungsgrenzen verschiedener ZfP-Verfahren dar.

  14. 33. MPA seminar 2007. Proceedings; 33. MPA Seminar 2007. Vortraege

    Energy Technology Data Exchange (ETDEWEB)



    In a single PDF file, the CD-ROM contains all papers presented at the 33. MPA Seminar in Stuttgart. Subjects: Life management of nuclear facilities: Materials, integrity analysis, fracture mechanics; Behaviour of welds; Material development and qualification for power plants with enhanced steam parameters; monitoring concepts (e.g. state monitoring of components, test concepts); Design of pipelines with flanged joints; Assembly and joining technology for flanged joints; Non-destructive testing in plant engineering; Materials for new nuclear reactors. Many of the papers are available as separate items in this database. [German] In einer einzigen PDF-Datei sind auf der CD-ROM alle Vortraege enthalten, die auf dem 33. MPA-Seminar in Stuttgart gehalten wurden. Die Themenschwerpunkte sind: Lebensdauermanagement kerntechnischer Anlagen - Werkstoffverhalten, Integritaetsanalyse, Bruchmechanik; Verhalten von Schweissverbindungen; Werkstoffentwicklung und -qualifizierung fuer Kraftwerke mit erhoehten Dampfparametern; Ueberwachungskonzepte (z.B. Zustandsueberwachung von Komponenten, Pruefkonzepte); Auslegung von Rohrleitungen mit Flanschverbindungen; Montage- und Dichtungstechnik fuer Flanschverbindungen; Zerstoerungsfreie Pruefung in der Anlagentechnik; Werkstoffe fuer neue Kernreaktoren. Ein grosser Teil der Vortraege wurde separat aufgenommen.

  15. The quality assurance topical report - International co-operation for manufacturing of nuclear packages

    International Nuclear Information System (INIS)

    Anderson, R.; Bonifacio, A.; Kingsley, K.; Bittner, R.; Ankrum, G.T.; Wieser, K.E.


    The paper addresses the manufacturing quality assurance (QA) concerns associated with fabrication of radioactive material packaging in one country for use in another. A case study is discussed which shows a method of implementation that has been successfully practised for casks fabricated in the Federal Republic of Germany (FRG) for use in the USA. The casks utilized are the Castor series, which are fabricated in the FRG from ductile cast iron. Although international efforts have helped to standardize QA measures, detailed implementation can vary from country to country. A means of co-ordinating requirements between countries was required. The vehicle used to accomplish this was the Quality Assurance Topical Report (QATR), which is an administrative QA document that clearly defines the roles and responsibilities of all affected organizations. The close correlation of requirements is documented in the QATR. In addition, a programme is described which permits the USA competent authority to depend on the FRG authority for inspection and enforcement procedures. It is of great importance that the fabrication and inspection of any component important to safety be properly controlled and documented. The USA requirements for these QA measures are enforced by the Nuclear Regulatory Commission (NRC). Enforcement of applicable regulations in the FRG is the responsibility of the competent authority, Bundesanstalt fuer Materialpruefung (BAM). As a part of the development of this QATR, NRC and BAM representatives met to establish a mutual understanding of QA. As a result, the QATR concept has been approved by both BAM and the NRC. A problem with manufacturing equipment in a foreign country is the need to periodically send company representatives to witness hold points and to review manufacturing records. The methods described in the QATR provide a practical and workable alternative by permitting the BAM to serve as the purchaser (or utility) agent. The utility is only required

  16. The Transcriptional Repressive Activity of KRAB Zinc Finger Proteins Does Not Correlate with Their Ability to Recruit TRIM28.

    Directory of Open Access Journals (Sweden)

    Kristin E Murphy

    Full Text Available KRAB domain Zinc finger proteins are one of the most abundant families of transcriptional regulators in higher vertebrates. The prevailing view is that KRAB domain proteins function as potent transcriptional repressors by recruiting TRIM28 and promoting heterochromatin spreading. However, the extent to which all KRAB domain proteins are TRIM28-dependent transcriptional repressors is currently unclear. Our studies on mouse ZFP568 revealed that TRIM28 recruitment by KRAB domain proteins is not sufficient to warrant transcriptional repressive activity. By using luciferase reporter assays and yeast two-hybrid experiments, we tested the ability of ZFP568 and other mouse KRAB domain proteins to repress transcription and bind TRIM28. We found that some mouse KRAB domain proteins are poor transcriptional repressors despite their ability to recruit TRIM28, while others showed strong KRAB-dependent transcriptional repression, but no TRIM28 binding. Together, our results show that the transcriptional repressive activity of KRAB-ZNF proteins does not correlate with their ability to recruit TRIM28, and provide evidence that KRAB domains can regulate transcription in a TRIM28-independent fashion. Our findings challenge the current understanding of the molecular mechanisms used by KRAB domain proteins to control gene expression and highlight that a high percentage of KRAB domain proteins in the mouse genome differ from the consensus KRAB sequence at amino acid residues that are critical for TRIM28 binding and/or repressive activity.

  17. Disruption of murine mp29/Syf2/Ntc31 gene results in embryonic lethality with aberrant checkpoint response.

    Directory of Open Access Journals (Sweden)

    Chia-Hsin Chen

    Full Text Available Human p29 is a putative component of spliceosomes, but its role in pre-mRNA is elusive. By siRNA knockdown and stable overexpression, we demonstrated that human p29 is involved in DNA damage response and Fanconi anemia pathway in cultured cells. In this study, we generated p29 knockout mice (mp29(GT/GT using the mp29 gene trap embryonic stem cells to study the role of mp29 in DNA damage response in vivo. Interruption of mp29 at both alleles resulted in embryonic lethality. Embryonic abnormality occurred as early as E6.5 in mp29(GT/GT mice accompanied with decreased mRNA levels of α-tubulin and Chk1. The reduction of α-tubulin and Chk1 mRNAs is likely due to an impaired post-transcriptional event. An aberrant G2/M checkpoint was found in mp29 gene trap embryos when exposed to aphidicolin and UV light. This embryonic lethality was rescued by crossing with mp29 transgenic mice. Additionally, the knockdown of zfp29 in zebrafish resulted in embryonic death at 72 hours of development postfertilization (hpf. A lower level of acetylated α-tubulin was also observed in zfp29 morphants. Together, these results illustrate an indispensable role of mp29 in DNA checkpoint response during embryonic development.

  18. Targeted disruption in mice of a neural stem cell-maintaining, KRAB-Zn finger-encoding gene that has rapidly evolved in the human lineage.

    Directory of Open Access Journals (Sweden)

    Huan-Chieh Chien

    Full Text Available Understanding the genetic basis of the physical and behavioral traits that separate humans from other primates is a challenging but intriguing topic. The adaptive functions of the expansion and/or reduction in human brain size have long been explored. From a brain transcriptome project we have identified a KRAB-Zn finger protein-encoding gene (M003-A06 that has rapidly evolved since the human-chimpanzee separation. Quantitative RT-PCR analysis of different human tissues indicates that M003-A06 expression is enriched in the human fetal brain in addition to the fetal heart. Furthermore, analysis with use of immunofluorescence staining, neurosphere culturing and Western blotting indicates that the mouse ortholog of M003-A06, Zfp568, is expressed mainly in the embryonic stem (ES cells and fetal as well as adult neural stem cells (NSCs. Conditional gene knockout experiments in mice demonstrates that Zfp568 is both an NSC maintaining- and a brain size-regulating gene. Significantly, molecular genetic analyses show that human M003-A06 consists of 2 equilibrated allelic types, H and C, one of which (H is human-specific. Combined contemporary genotyping and database mining have revealed interesting genetic associations between the different genotypes of M003-A06 and the human head sizes. We propose that M003-A06 is likely one of the genes contributing to the uniqueness of the human brain in comparison to other higher primates.

  19. Custom-Designed Molecular Scissors for Site-Specific Manipulation of the Plant and Mammalian Genomes (United States)

    Kandavelou, Karthikeyan; Chandrasegaran, Srinivasan

    Zinc finger nucleases (ZFNs) are custom-designed molecular scissors, engineered to cut at specific DNA sequences. ZFNs combine the zinc finger proteins (ZFPs) with the nonspecific cleavage domain of the FokI restriction enzyme. The DNA-binding specificity of ZFNs can be easily altered experimentally. This easy manipulation of the ZFN recognition specificity enables one to deliver a targeted double-strand break (DSB) to a genome. The targeted DSB stimulates local gene targeting by several orders of magnitude at that specific cut site via homologous recombination (HR). Thus, ZFNs have become an important experimental tool to make site-specific and permanent alterations to genomes of not only plants and mammals but also of many other organisms. Engineering of custom ZFNs involves many steps. The first step is to identify a ZFN site at or near the chosen chromosomal target within the genome to which ZFNs will bind and cut. The second step is to design and/or select various ZFP combinations that will bind to the chosen target site with high specificity and affinity. The DNA coding sequence for the designed ZFPs are then assembled by polymerase chain reaction (PCR) using oligonucleotides. The third step is to fuse the ZFP constructs to the FokI cleavage domain. The ZFNs are then expressed as proteins by using the rabbit reticulocyte in vitro transcription/translation system and the protein products assayed for their DNA cleavage specificity.

  20. Immunosuppressive potential of bardoxolone methyl using a modified murine local lymph node assay (LLNA). (United States)

    Kitsukawa, Mika; Tsuchiyama, Hiromi; Maeda, Akihisa; Oshida, Keiyu; Miyamoto, Yohei


    2-Cyano-3, 12-dioxooleana-1, 9-dien-28-oic acid methyl ester (CDDO-Me; bardoxolone methyl) is one of the synthetic oleanane triterpenoids (SOs). It is known that it is the strongest Nrf2/ARE signaling inducer of SOs and slightly inhibits immune response. Little was known about the immunomodulatory action of CDDO-Me in vivo. We assessed its immunosuppressive potential by using the modified mouse lymph node assay (LLNA) including immunosuppression-related gene expression analysis. In the modified LLNA, CDDO-Me showed a significant decrease in lymph node weight and changes in expressions of the immunosuppression-related genes, Zfp459 and Fmo2. It has been already reported that a decrease in lymph node weight was induced by several types of immunosuppressive chemicals such as calcineurin inhibitors, antimetabolites, steroids, and alkylators. In addition, changes in Zfp459 and Fmo2 expression was reported in response after only treatment of antimetabolites. From these results, CDDO-Me is considered to have an immunosuppressive action and similar mechanism to antimetabolites.

  1. Global and stage specific patterns of Krüppel-associated-box zinc finger protein gene expression in murine early embryonic cells.

    Directory of Open Access Journals (Sweden)

    Andrea Corsinotti

    Full Text Available Highly coordinated transcription networks orchestrate the self-renewal of pluripotent stem cell and the earliest steps of mammalian development. KRAB-containing zinc finger proteins represent the largest group of transcription factors encoded by the genomes of higher vertebrates including mice and humans. Together with their putatively universal cofactor KAP1, they have been implicated in events as diverse as the silencing of endogenous retroelements, the maintenance of imprinting and the pluripotent self-renewal of embryonic stem cells, although the genomic targets and specific functions of individual members of this gene family remain largely undefined. Here, we first generated a list of Ensembl-annotated KRAB-containing genes encoding the mouse and human genomes. We then defined the transcription levels of these genes in murine early embryonic cells. We found that the majority of KRAB-ZFP genes are expressed in mouse pluripotent stem cells and other early progenitors. However, we also identified distinctively cell- or stage-specific patterns of expression, some of which are pluripotency-restricted. Finally, we determined that individual KRAB-ZFP genes exhibit highly distinctive modes of expression, even when grouped in genomic clusters, and that these cannot be correlated with the presence of prototypic repressive or activating chromatin marks. These results pave the way to delineating the role of specific KRAB-ZFPs in early embryogenesis.

  2. On-line monitoring and inservice inspection in codes; Betriebsueberwachung und wiederkehrende Pruefungen in den Regelwerken

    Energy Technology Data Exchange (ETDEWEB)

    Bartonicek, J.; Zaiss, W. [Gemeinschaftskernkraftwerk Neckar GmbH, Neckarwestheim (Germany); Bath, H.R. [Bundesamt fuer Strahlenschutz, Salzgitter (Germany). Geschaeftsstelle des Kerntechnischen Ausschusses (KTA)


    Betriebsueberwachung und wiederkehrende Pruefungen in angemessenem Umfang. In auslaendischen kerntechnischen Regelwerken erfolgt die Absicherung im Betrieb hauptsaechlich durch zerstoerungsfreie wiederkehrende Pruefungen, wobei der Umfang und die Pruefintervalle vorwiegend vom ASME-Code, Section XI abgeleitet wurden. (orig.)

  3. Training to NDT construction experts (ZFPBau). Developments in recent years; Ausbildung zum ZfPBau-Experten. Entwicklungen der letzten Jahre

    Energy Technology Data Exchange (ETDEWEB)

    Taffe, Alexander [Hochschule fuer Technik und Wirtschaft, Berlin (Germany); Feistkorn, Sascha [SVTI Schweizerischer Verein fuer technische Inspektionen, Wallisellen (Switzerland). Nuklearinspektorat; Dauberschmidt, Christoph [Hochschule Muenchen (Germany)


    Classical destructive test methods are fixed part of industrial sectors for decades such as aerospace, automotive, railway and power plants. A high degree of standardization of procedures, and training in form of an ISO standard are present, but not in non-destructive testing of civil engineering (ZfPBau). Except for the rebound hammer NDT methods in civil engineering are not standardized. Also of personnel qualifications can be found in regulations very few requirements, although it is obvious that this is an indispensable prerequisite for the proper application of the procedures and reliable inspection results. In this contribution experiences with a presented training concept in which the construction inspector will trained to DIN 1076 in a two-day course. Here are the necessary conditions worked out, such as the creation of suitable test bodies or the definition of necessary course content for establishing training courses. [German] Klassische zerstoerungsfreie Pruefverfahren sind seit Jahrzehnten fester Bestandteil industrieller Bereiche wie Luftfahrt, Automotive, Eisenbahn und Kraftwerke. Ein hoher Normungsgrad der Verfahren und Ausbildungsstandards in Form einer ISO-Norm liegen vor. Bei der zerstoerungsfreien Pruefung im Bauwesen (ZfPBau) ist das anders. Bis auf den Rueckprallhammer sind ZfPBau-Verfahren nicht genormt. Auch zur Personalqualifikation finden sich in Regelwerken nur sehr wenige Anforderungen, obwohl es offensichtlich ist, dass dies eine unverzichtbare Voraussetzung fuer die richtige Anwendung der Verfahren und zuverlaessige Pruefaussagen ist. In diesem Beitrag werden Erfahrungen mit einem Ausbildungskonzept vorgestellt, bei dem Bauwerkspruefer nach DIN 1076 in einem zweitaegigen Lehrgang ausgebildet werden. Dabei werden die notwendigen Voraussetzungen heraus gearbeitet, wie z.B. die Erstellung von geeigneten Testkoerpern oder die Festlegung notwendiger Kursinhalte, die zum Etablieren von Ausbildungskursen erforderlich sind.

  4. Residual stress analysis in carbon fiber-reinforced SiC ceramics; Eigenspannungsanalyse in kohlenstoffaserverstaerkten SiC-Keramiken

    Energy Technology Data Exchange (ETDEWEB)

    Broda, M.


    Systematic residual stress analyses are reported, carried out in long-fiber reinforced SiC ceramics. The laminated C{sub fiber}/SiC{sub matrix} specimens used were prepared by polymer pyrolysis, and the structural component specimens used are industrial products. Various diffraction methods have been applied for non-destructive evaluation of residual stress fields, so as to completely detect the residual stresses and their distribution in the specimens. The residual stress fields at the surface ({mu}m) have been measured using characteristic X-radiation and applying the sin {sup 2}{psi} method as well as the scatter vector method. For residual stress field analysis in the mass volume (cm), neutron diffraction has been applied. The stress fields in the fiber layers (approx. 250{mu}m) have been measured as a function of their location within the laminated composite by using an energy-dispersive method and synchrotron radiation. By means of the systematic, process-accompanying residual stress and phase analyses, conclusions can be drawn as to possible approaches for optimization of fabrication parameters. (orig./CB) [Deutsch] Im Rahmen der Arbeit werden systematische Eigenspannungsanalysen an langfaserverstaerkten SiC-Keramiken durchgefuehrt. Hierbei werden polymerpyrolytisch abgeleitete, laminierte C{sub Faser}/SiC{sub Matrix} Proben und Bauteile untersucht, welche industriell gefertigt wurden. Fuer die zerstoerungsfreie Eigenspannungsermittlung kommen verschiedene Beugungsverfahren zum Einsatz. Dadurch kann die Eigenspannungsverteilung in diesen Proben vollstaendig erfasst werden, d.h. der Eigenspannungszustand im Oberflaechenbereich ({mu}m) wird mit Hilfe charakteristischer Roentgenstrahlung unter Nutzung der sin{sup 2}{psi}-Methode als auch der Streuvektor-Methode beschrieben. Fuer die Analyse der Eigenspannungen im Volumen (cm) wird die Neutronenbeugung herangezogen. Um den Spannungszustand in den einzelnen Fasermatten (ca. 250 {mu}m) in Abhaengigkeit ihrer Lage

  5. The industrial application of fracture mechanics concepts discussed at the background of international standards and guidelines; Die industrielle Anwendung bruchmechanischer Konzepte vor dem Hintergrund internationaler Bewertungsvorschriften und Regelwerke

    Energy Technology Data Exchange (ETDEWEB)

    Zerbst, U. [GKSS-Forschungszentrum Geesthacht GmbH (Germany). Inst. fuer Werkstofforschung; Langenberg, P. [Ingenieurbuero fuer Werkstofftechnik, Aachen (Germany)


    Many features from the background for an intensified application of fracture mechanics concepts in many industries world-wide. These include requirements for a permanent increase of the level of performance of technical components and structures by the introduction of new materials, joining technologies and design principles, the problem of ageing components and life extension, an increased emphasis on non-destructive in-service inspection combined with improved NDT techniques, and also a number of failure events caused by fatigue and fracture The aim of the present paper is to give a brief state-of-the-art review on how fracture mechanics is applied in different industrial branches today. This is based on standards and guidelines in the aerospace industry, in the nuclear and fossil power generation, in the chemical and petrochemical and the pipeline industry, in civil engineering, offshore technique and other fields. Based on the review an outlook is given on a future development that would be reasonable and desirable from the point of view of a basically unified philosophy of fracture mechanics application. (orig.) [German] Die Erhoehung der Leistungsparameter vieler Maschinen und Anlagen verbunden mit dem Einsatz neuer Werkstoffe, Fuegeverfahren und Konstruktionsprinzipien, der Betrieb vieler Strukturen ueber ihre projektierte Lebensdauer hinaus, technische Verbesserungen und ein durchgaengigerer Einsatz zerstoerungsfreier Defektpruefverfahren, aber auch immer wieder einmal auftretende Schadensfaelle bilden den Hintergrund fuer die zunehmende Nutzung bruchmechanischer Bewertungsvorschriften in der industriellen Praxis. Die vorliegende Studie zieht eine momentane Bilanz dieser Entwicklung am Beispiel von Fachbereichsstandards der Luft- und Raumfahrtindustrie, der konventionellen und Kernkrafttechnik, der Chemie und Petrochemie, der Pipelineindustrie, des Stahlbaus, der Offshore-Technik und anderer Bereiche. Ausgehend von dieser Bestandsaufnahme wird ein Ausblick

  6. TomoWELD. Precise detection of weld defects; TomoWELD. Defekte in Schweissnaehten praezise erkennen

    Energy Technology Data Exchange (ETDEWEB)

    Walter, David [Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin (Germany)


    katastrophale Folgen haben. Daher muessen in Kernkraftwerken ausnahmslos alle Schweissnaehte ueberprueft werden. Dafuer zugelassene zerstoerungsfreie Methoden verwenden Ultraschall und Roentgenstrahlung. Die an der BAM entwickelte Technik heisst TomoWELD.

  7. Inherent dynamics of head domain correlates with ATP-recognition of P2X4 receptors: insights gained from molecular simulations.

    Directory of Open Access Journals (Sweden)

    Li-Dong Huang

    Full Text Available P2X receptors are ATP-gated ion channels involved in many physiological functions, and determination of ATP-recognition (AR of P2X receptors will promote the development of new therapeutic agents for pain, inflammation, bladder dysfunction and osteoporosis. Recent crystal structures of the zebrafish P2X4 (zfP2X4 receptor reveal a large ATP-binding pocket (ABP located at the subunit interface of zfP2X4 receptors, which is occupied by a conspicuous cluster of basic residues to recognize triphosphate moiety of ATP. Using the engineered affinity labeling and molecular modeling, at least three sites (S1, S2 and S3 within ABP have been identified that are able to recognize the adenine ring of ATP, implying the existence of at least three distinct AR modes in ABP. The open crystal structure of zfP2X4 confirms one of three AR modes (named AR1, in which the adenine ring of ATP is buried into site S1 while the triphosphate moiety interacts with clustered basic residues. Why architecture of ABP favors AR1 not the other two AR modes still remains unexplored. Here, we examine the potential role of inherent dynamics of head domain, a domain involved in ABP formation, in AR determinant of P2X4 receptors. In silico docking and binding free energy calculation revealed comparable characters of three distinct AR modes. Inherent dynamics of head domain, especially the downward motion favors the preference of ABP for AR1 rather than AR2 and AR3. Along with the downward motion of head domain, the closing movement of loop139-146 and loop169-183, and structural rearrangements of K70, K72, R298 and R143 enabled ABP to discriminate AR1 from other AR modes. Our observations suggest the essential role of head domain dynamics in determining AR of P2X4 receptors, allowing evaluation of new strategies aimed at developing specific blockers/allosteric modulators by preventing the dynamics of head domain associated with both AR and channel activation of P2X4 receptors.

  8. ZifBASE: a database of zinc finger proteins and associated resources

    Directory of Open Access Journals (Sweden)

    Punetha Ankita


    Full Text Available Abstract Background Information on the occurrence of zinc finger protein motifs in genomes is crucial to the developing field of molecular genome engineering. The knowledge of their target DNA-binding sequences is vital to develop chimeric proteins for targeted genome engineering and site-specific gene correction. There is a need to develop a computational resource of zinc finger proteins (ZFP to identify the potential binding sites and its location, which reduce the time of in vivo task, and overcome the difficulties in selecting the specific type of zinc finger protein and the target site in the DNA sequence. Description ZifBASE provides an extensive collection of various natural and engineered ZFP. It uses standard names and a genetic and structural classification scheme to present data retrieved from UniProtKB, GenBank, Protein Data Bank, ModBase, Protein Model Portal and the literature. It also incorporates specialized features of ZFP including finger sequences and positions, number of fingers, physiochemical properties, classes, framework, PubMed citations with links to experimental structures (PDB, if available and modeled structures of natural zinc finger proteins. ZifBASE provides information on zinc finger proteins (both natural and engineered ones, the number of finger units in each of the zinc finger proteins (with multiple fingers, the synergy between the adjacent fingers and their positions. Additionally, it gives the individual finger sequence and their target DNA site to which it binds for better and clear understanding on the interactions of adjacent fingers. The current version of ZifBASE contains 139 entries of which 89 are engineered ZFPs, containing 3-7F totaling to 296 fingers. There are 50 natural zinc finger protein entries ranging from 2-13F, totaling to 307 fingers. It has sequences and structures from literature, Protein Data Bank, ModBase and Protein Model Portal. The interface is cross linked to other public

  9. Improvement and qualification of ultrasonic testing of dissimilar welds in the primary circuit of NPPs; Verbesserung und Qualifizierung der Ultraschallpruefung von Mischnaehten im Primaerkreis von KKW

    Energy Technology Data Exchange (ETDEWEB)

    Mitzscherling, Steffen; Barth, Enrico; Homann, Tobias; Prager, Jens [Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin (Germany); Goetschel, Sebastian; Weiser, Martin [Konrad-Zuse-Zentrum fuer Informationstechnik Berlin (ZIB) (Germany)


    The austenitic and dissimilar welds found in the primary circuit of nuclear power plants are not only extremely relevant to safety but also place very high demands on material testing. In addition to limited accessibility, the macroscopic structure of the weld seam is of paramount importance for ultrasound testing. In order to reliably determine material errors in position and size, the grain orientations and the elastic constants of the anisotropic weld bead structure must be known. The following work steps are used for the imaging representation of possible material defects: First, the weld seam is sounded in order to be able to determine important weld seam parameters, such as, for example, the grain orientation, using an inverse method. On the basis of these parameters, the sound paths are simulated in the next step by means of raytracing (RT). Finally, this RT simulation is assigned the measurement data (A-scans) from different transmitter and receiver positions and superimposed according to the Synthetic Aperature Focusing Technique (SAFT) method. The combination of inverse process, RT and SAFT also ensures a correct visualization of the faults in anisotropic materials. We explain these three methods and present the test arrangement of test specimens with artificial test errors. Measurement data as well as their evaluation are compared with the results of a CIVA simulation. [German] Die im Primaerkreislauf von Kernkraftwerken anzutreffenden austenitischen Schweiss- und Mischnaehte sind nicht nur extrem sicherheitsrelevant, sondern stellen auch sehr hohe Anforderungen an die Materialpruefung. Neben der eingeschraenkten Zugaenglichkeit ist das makroskopische Gefuege der Schweissnaht fuer die Pruefung mit Ultraschall von hoechster Bedeutung. Um Materialfehler zuverlaessig in Position und Groesse bestimmen zu koennen, muessen die Kornorientierungen und die elastischen Konstanten des anisotropen Schweissnahtgefueges bekannt sein. Fuer die bildgebende Darstellung

  10. Glucocorticoid and cytokine crosstalk: Feedback, feedforward, and co-regulatory interactions determine repression or resistance. (United States)

    Newton, Robert; Shah, Suharsh; Altonsy, Mohammed O; Gerber, Antony N


    Inflammatory signals induce feedback and feedforward systems that provide temporal control. Although glucocorticoids can repress inflammatory gene expression, glucocorticoid receptor recruitment increases expression of negative feedback and feedforward regulators, including the phosphatase, DUSP1, the ubiquitin-modifying enzyme, TNFAIP3, or the mRNA-destabilizing protein, ZFP36. Moreover, glucocorticoid receptor cooperativity with factors, including nuclear factor-κB (NF-κB), may enhance regulator expression to promote repression. Conversely, MAPKs, which are inhibited by glucocorticoids, provide feedforward control to limit expression of the transcription factor IRF1, and the chemokine, CXCL10. We propose that modulation of feedback and feedforward control can determine repression or resistance of inflammatory gene expression toglucocorticoid. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  11. 2017 Annual meeting of the German Society for Non-destructive Testing (DGZfP). Report; DGZfP-Jahrestagung 2017. Berichtsband

    Energy Technology Data Exchange (ETDEWEB)



    The proceedings contain lectures with the following main Standards and regulations; Material characterization; Production monitoring; Ultrasound procedures; Simulation; Various NDT procedures; Automotive; Air ultrasound; Algorithms / simulation; ZfP knowledge yesterday, today and tomorrow; UT Phased Array - Total Focusing Method; Composite materials; Transportation - Railway; Wooden structures; Thermography; Pressure equipment; Computed Tomography; Defect detection / acoustic procedures; Reliability (POD); Composite materials - methods; Transportation - Aviation; Condition monitoring; Microwaves and terahertz. Two lectures were recorded separately for this database. [German] Dieser Tagungsband enthaelt Vortraege mit folgenden Themenschwerpunkten: Normen und Regelwerke, Materialcharakterisierung, Fertigungsueberwachung, Ultraschallverfahren, Simulation, Diverse ZfP-Verfahren, Automotive, Luftultraschall, Algorithmen/Simulation, ZfP-Wissen gestern, heute und morgen, UT Phased Array - Total Focusing Method, Verbundwerkstoffe, Verkehrswesen - Bahn, Holzstrukturen, Thermographie, Druckgeraete, Computertomographie, Defektnachweis/ Akustische Verfahren, Reliability (POD), Verbundwerkstoffe - Verfahren, Verkehrswesen - Luftfahrt, Zustandsueberwachung, Mikrowellen und TeraHertz. Zwei Vortraege wurden fuer diese Datenbank separat aufgenommen.

  12. Microarray-based screening of differentially expressed genes in glucocorticoid-induced avascular necrosis (United States)

    Huang, Gangyong; Wei, Yibing; Zhao, Guanglei; Xia, Jun; Wang, Siqun; Wu, Jianguo; Chen, Feiyan; Chen, Jie; Shi, Jingshen


    The underlying mechanisms of glucocorticoid (GC)-induced avascular necrosis of the femoral head (ANFH) have yet to be fully understood, in particular the mechanisms associated with the change of gene expression pattern. The present study aimed to identify key genes with a differential expression pattern in GC-induced ANFH. E-MEXP-2751 microarray data were downloaded from the ArrayExpress database. Differentially expressed genes (DEGs) were identified in 5 femoral head samples of steroid-induced ANFH rats compared with 5 placebo-treated rat samples. Gene Ontology (GO) and pathway enrichment analyses were performed upon these DEGs. A total 93 DEGs (46 upregulated and 47 downregulated genes) were identified in GC-induced ANFH samples. These DEGs were enriched in different GO terms and pathways, including chondrocyte differentiation and detection of chemical stimuli. The enrichment map revealed that skeletal system development was interconnected with several other GO terms by gene overlap. The literature mined network analysis revealed that 5 upregulated genes were associated with femoral necrosis, including parathyroid hormone receptor 1 (PTHR1), vitamin D (1,25-Dihydroxyvitamin D3) receptor (VDR), collagen, type II, α1, proprotein convertase subtilisin/kexin type 6 and zinc finger protein 354C (ZFP354C). In addition, ZFP354C and VDR were identified to transcription factors. Furthermore, PTHR1 was revealed to interact with VDR, and α-2-macroglobulin (A2M) interacted with fibronectin 1 (FN1) in the PPI network. PTHR1 may be involved in GC-induced ANFH via interacting with VDR. A2M may also be involved in the development of GC-induced ANFH through interacting with FN1. An improved understanding of the molecular mechanisms underlying GC-induced ANFH may provide novel targets for diagnostics and therapeutic treatment. PMID:28393228

  13. Microarray‑based screening of differentially expressed genes in glucocorticoid‑induced avascular necrosis. (United States)

    Huang, Gangyong; Wei, Yibing; Zhao, Guanglei; Xia, Jun; Wang, Siqun; Wu, Jianguo; Chen, Feiyan; Chen, Jie; Shi, Jingshen


    The underlying mechanisms of glucocorticoid (GC)‑induced avascular necrosis of the femoral head (ANFH) have yet to be fully understood, in particular the mechanisms associated with the change of gene expression pattern. The present study aimed to identify key genes with a differential expression pattern in GC‑induced ANFH. E‑MEXP‑2751 microarray data were downloaded from the ArrayExpress database. Differentially expressed genes (DEGs) were identified in 5 femoral head samples of steroid‑induced ANFH rats compared with 5 placebo‑treated rat samples. Gene Ontology (GO) and pathway enrichment analyses were performed upon these DEGs. A total 93 DEGs (46 upregulated and 47 downregulated genes) were identified in GC‑induced ANFH samples. These DEGs were enriched in different GO terms and pathways, including chondrocyte differentiation and detection of chemical stimuli. The enrichment map revealed that skeletal system development was interconnected with several other GO terms by gene overlap. The literature mined network analysis revealed that 5 upregulated genes were associated with femoral necrosis, including parathyroid hormone receptor 1 (PTHR1), vitamin D (1,25‑Dihydroxyvitamin D3) receptor (VDR), collagen, type II, α1, proprotein convertase subtilisin/kexin type 6 and zinc finger protein 354C (ZFP354C). In addition, ZFP354C and VDR were identified to transcription factors. Furthermore, PTHR1 was revealed to interact with VDR, and α‑2‑macroglobulin (A2M) interacted with fibronectin 1 (FN1) in the PPI network. PTHR1 may be involved in GC‑induced ANFH via interacting with VDR. A2M may also be involved in the development of GC‑induced ANFH through interacting with FN1. An improved understanding of the molecular mechanisms underlying GC‑induced ANFH may provide novel targets for diagnostics and therapeutic treatment.

  14. C/EBPβ Mediates Growth Hormone-Regulated Expression of Multiple Target Genes (United States)

    Cui, Tracy X.; Lin, Grace; LaPensee, Christopher R.; Calinescu, Anda-Alexandra; Rathore, Maanjot; Streeter, Cale; Piwien-Pilipuk, Graciela; Lanning, Nathan; Jin, Hui; Carter-Su, Christin; Qin, Zhaohui S.


    Regulation of c-Fos transcription by GH is mediated by CCAAT/enhancer binding protein β (C/EBPβ). This study examines the role of C/EBPβ in mediating GH activation of other early response genes, including Cyr61, Btg2, Socs3, Zfp36, and Socs1. C/EBPβ depletion using short hairpin RNA impaired responsiveness of these genes to GH, as seen for c-Fos. Rescue with wild-type C/EBPβ led to GH-dependent recruitment of the coactivator p300 to the c-Fos promoter. In contrast, rescue with C/EBPβ mutated at the ERK phosphorylation site at T188 failed to induce GH-dependent recruitment of p300, indicating that ERK-mediated phosphorylation of C/EBPβ at T188 is required for GH-induced recruitment of p300 to c-Fos. GH also induced the occupancy of phosphorylated C/EBPβ and p300 on Cyr61, Btg2, and Socs3 at predicted C/EBP-cAMP response element-binding protein motifs in their promoters. Consistent with a role for ERKs in GH-induced expression of these genes, treatment with U0126 to block ERK phosphorylation inhibited their GH-induced expression. In contrast, GH-dependent expression of Zfp36 and Socs1 was not inhibited by U0126. Thus, induction of multiple early response genes by GH in 3T3-F442A cells is mediated by C/EBPβ. A subset of these genes is regulated similarly to c-Fos, through a mechanism involving GH-stimulated ERK 1/2 activation, phosphorylation of C/EBPβ, and recruitment of p300. Overall, these studies suggest that C/EBPβ, like the signal transducer and activator of transcription proteins, regulates multiple genes in response to GH. PMID:21292824

  15. Calibration and characterisation of solar cells: Comparison of pulsed and steady-state simulation of sunlight; Kalibrierung und Charakterisierung von Solarzellen: Vergleich von gepulster und stationaerer Sonnensimulation

    Energy Technology Data Exchange (ETDEWEB)

    Metzdorf, J; Kessler, F R; Bentlage, V; Holstenberg, H C; Sperling, A; Winter, S; Wittchen, T


    lassen sich mit einem stationaeren Simulator bei vertretbarem technischen und wirtschaftlichen Aufwand nicht erzeugen. Flasher-Kalibrierungen fuehren jedoch oft zu Fehlern insbesondere bei hocheffizienten Solarzellen, verursacht durch Verzerrungen der transient gemessenen Strom-Spannungs-Kennlinien. Die daraus folgenden Kennlinienaufspaltungen wurden in Abhaengigkeit von Solarzellentechnologie und Messbedingungen systematisch untersucht, dokumentiert und modellmaessig simuliert. Auf der Basis dieser Grundsatzanalyse lassen sich die prinzipiell unvermeidbaren transienten Stoereffekte nicht nur erklaeren sondern auch vollstaendig korrigieren. Dazu wurde ein einfaches Korrekturverfahren fuer die Umrechnung in die unverzerrte Kennlinie entwickelt, das nur zwei bestimmte Dunkelkennlinien aus Hilfsmessungen benoetigt. Daneben wurden Detailverbesserungen fuer die Kalibrierung von Referenzsolarzellen erreicht, Prototypen grossflaechiger Referenzzellen entwickelt sowie neue und weiterentwickelte transiente und quasistationaere Laborverfahren zur optoelektronischen Charakterisierung von Solarzellen getestet, mit denen Stoerstelleneigenschaften ueberwiegend zerstoerungsfrei und z.T. orts- bzw. tiefenaufgeloest untersucht werden koennen. (orig.)

  16. Residual stresses in microwave sintered Ni/8Y-ZrO{sub 2} and NiCr8020/8Y-ZrO{sub 2} gradient materials; Eigenspannungen in mikrowellengesinterten Ni/8Y-ZrO{sub 2} und NiCr8020/8Y-ZrO{sub 2} Gradientenwerkstoffen

    Energy Technology Data Exchange (ETDEWEB)

    Dantz, D.


    The connection between the residual macro- and microstresses and the macro- and micromechanical properties of pressureless microwave-sintered NiCr8020/8Y-ZrO{sub 2}, or Ni/8Y-ZrO{sub 2}, respectively, layered composites with gradient intermediate layers will be analysed by way of experimental methods. Non-destructive residual stress analyses will be done using various diffraction methods. Apart from conventional methods such as the sin{sup 2}{psi} method, or neutron scattering, specific analytical methods will be applied for examining the gradient sintered composites. The residual stress gradients found in the surface regions of specimens will be analysed with the scattering vector method. For examination of residual stress distributions in the core layers of the sintered specimens, an energy-dispersive technique with very high local resolution will be used, applying high-energy synchrotron radiation. The technique permits an analysis of residual stress distributions even in a homogeneous single layer of a gradient composite. In addition, line profile analyses will be made, and optical and electron-microscopical methods will be used for microstructure analyses. (orig./CB) [German] Im Rahmen dieser Arbeit wurde der Zusammenhang zwischen den Makro- und Mikroeigenspannungen und den makro- und mikromechanischen Eigenschaften an drucklos mikrowellengesinterten NiCr8020/8Y-ZrO{sub 2}- bzw. Ni/8Y-ZrO{sub 2}-Schichtverbundsystemen mit gradierter Zwischenschicht mit Hilfe experimenteller Methoden systematisch untersucht. Die Eigenspannungsanalysen erfolgten zerstoerungsfrei mit Hilfe unterschiedlicher Beugungsmethoden. Neben den konventionellen Methoden, wie dem sin{sup 2}{psi}-Verfahren oder auch der Neutronenbeugung, wurden zur Untersuchung der gradiert zusammengesetzten Sinterkoerper spezielle Analyseverfahren eingesetzt. Die im probenoberflaechennahen Bereich vorliegenden Eigenspannungsgradienten wurden mit Hilfe des Streuvektorverfahrens analysiert. Fuer die

  17. Use of modeling and simulation in the planning, analysis and interpretation of ultrasonic testing; Einsatz von Modellierung und Simulation bei der Planung, Analyse und Interpretation von Ultraschallpruefungen

    Energy Technology Data Exchange (ETDEWEB)

    Algernon, Daniel [SVTI Schweizerischer Verein fuer technische Inspektionen, Wallisellen (Switzerland). ZfP-Labor; Grosse, Christian U. [Technische Univ. Muenchen (Germany). Lehrstuhl fuer Zerstoerungsfreie Pruefung


    Acoustic testing methods such as ultrasound and impact echo are an important tool in building diagnostics. The range includes thickness measurements, the representation of the internal component geometry as well as the detection of voids (gravel pockets), delaminations or possibly locating grouting faults in the interior of metallic cladding tubes of tendon ducts. Basically acoustic method for non-destructive testing (NDT) is based on the excitation of elastic waves that interact with the target object (e.g. to detect discontinuity in the component) at the acoustic interface. From the signal received at the component surface this interaction shall be detected and interpreted to draw conclusions about the presence of the target object, and optionally to determine its size and position (approximately). Although the basic underlying physical principles of the application of elastic waves in NDT are known, it can be complicated by complex relationships in the form of restricted access, component geometries, or the type and form of reflectors. To estimate the chances of success of a test is already often not trivial. These circumstances highlight the importance of using simulations that allow a theoretically sound basis for testing and allow easy optimizing test systems. The deployable simulation methods are varied. Common are in particular the finite element method, the Elasto Finite Integration Technique and semi-analytical calculation methods. [German] Akustische Pruefverfahren wie Ultraschall und Impact-Echo sind ein wichtiges Werkzeug der Bauwerksdiagnose. Das Einsatzspektrum beinhaltet Dickenmessungen, die Darstellung der inneren Bauteilgeometrie ebenso wie die Ortung von Kiesnestern, Delaminationen oder u.U. die Ortung von Verpressfehlern im Innern metallischer Huellrohre von Spannkanaelen. Grundsaetzlich beruhen akustische Verfahren zur Zerstoerungsfreien Pruefung (ZfP) auf der Anregung elastischer Wellen, die mit dem Zielobjekt (z. B. zu detektierende Ungaenze

  18. The ISTIMES project: a new integrated system for monitoring critical transport infrastructures interested by natural hazards (United States)

    Proto, Monica; Massimo, Bavusi; Francesco, Soldovieri


    , Eidgenoessische Materialpruefungs-und Forschungsanstalt (EMPA) from Switzerland, Laboratoire Central des Ponts et Chaussées (LCPC) from France, Lund University (ULUND) from Sweden, Tel Aviv University (TAU) from Israel, Territorial Data Elaboration (TDE) from Romania and Norsk Elektro Optikk (NEO) from Norway.

  19. The IDA-LIKE peptides IDL6 and IDL7 are negative modulators of stress responses in Arabidopsis thaliana. (United States)

    Vie, Ane Kjersti; Najafi, Javad; Winge, Per; Cattan, Ester; Wrzaczek, Michael; Kangasjärvi, Jaakko; Miller, Gad; Brembu, Tore; Bones, Atle M


    Small signalling peptides have emerged as important cell to cell messengers in plant development and stress responses. However, only a few of the predicted peptides have been functionally characterized. Here, we present functional characterization of two members of the IDA-LIKE (IDL) peptide family in Arabidopsis thaliana, IDL6 and IDL7. Localization studies suggest that the peptides require a signal peptide and C-terminal processing to be correctly transported out of the cell. Both IDL6 and IDL7 appear to be unstable transcripts under post-transcriptional regulation. Treatment of plants with synthetic IDL6 and IDL7 peptides resulted in down-regulation of a broad range of stress-responsive genes, including early stress-responsive transcripts, dominated by a large group of ZINC FINGER PROTEIN (ZFP) genes, WRKY genes, and genes encoding calcium-dependent proteins. IDL7 expression was rapidly induced by hydrogen peroxide, and idl7 and idl6 idl7 double mutants displayed reduced cell death upon exposure to extracellular reactive oxygen species (ROS). Co-treatment of the bacterial elicitor flg22 with IDL7 peptide attenuated the rapid ROS burst induced by treatment with flg22 alone. Taken together, our results suggest that IDL7, and possibly IDL6, act as negative modulators of stress-induced ROS signalling in Arabidopsis. © The Author 2017. Published by Oxford University Press on behalf of the Society for Experimental Biology.

  20. OsDOG, a gibberellin-induced A20/AN1 zinc-finger protein, negatively regulates gibberellin-mediated cell elongation in rice. (United States)

    Liu, Yaju; Xu, Yunyuan; Xiao, Jun; Ma, Qibin; Li, Dan; Xue, Zhen; Chong, Kang


    The A20/AN1 zinc-finger proteins (ZFPs) play pivotal roles in animal immune responses and plant stress responses. From previous gibberellin (GA) microarray data and A20/AN1 ZFP family member association, we chose Oryza sativa dwarf rice with overexpression of gibberellin-induced gene (OsDOG) to examine its function in the GA pathway. OsDOG was induced by gibberellic acid (GA(3)) and repressed by the GA-synthesis inhibitor paclobutrazol. Different transgenic lines with constitutive expression of OsDOG showed dwarf phenotypes due to deficiency of cell elongation. Additional GA(1) and real-time PCR quantitative assay analyses confirmed that the decrease of GA(1) in the overexpression lines resulted from reduced expression of GA3ox2 and enhanced expression of GA2ox1 and GA2ox3. Adding exogenous GA rescued the constitutive expression phenotypes of the transgenic lines. OsDOG has a novel function in regulating GA homeostasis and in negative maintenance of plant cell elongation in rice. Copyright © 2011 Elsevier GmbH. All rights reserved.

  1. Enhancement of adipogenesis and fibrogenesis in skeletal muscle of Wagyu compared with Angus cattle. (United States)

    Duarte, M S; Paulino, P V R; Das, A K; Wei, S; Serão, N V L; Fu, X; Harris, S M; Dodson, M V; Du, M


    Intramuscular fat and collagen content are major factors affecting beef quality, but mechanisms regulating intramuscular adipose and connective tissue deposition are far from clear. Japanese Wagyu cattle are well known for their extremely high marbling. The objective of this study was to evaluate intramuscular fat (IMF) and collagen deposition in the muscle of Wagyu compared with Angus cattle. Animals were managed under the same condition and slaughtered at an averaging 585 ± 12.1 kg of BW. Samples of sternomandibularis muscle were collected from Wagyu (n = 3) and Angus (n = 3) for molecular and histological investigations of adipogenesis and fibrogenesis. With exception of C/EBPβ (P = 0.2864), the expression of the adipogenic markers C/EBPα (P = 0.008), PPARγ (P = 0.028), and zip finger protein 423 (Zfp423; P = 0.047) in Wagyu were greater than in Angus muscle, which was consistent with greater IMF deposition in Wagyu (P Angus cattle. These results clearly show that both IMF and collagen contents are enhanced in Wagyu cattle and more adipogenic cells are detected in Wagyu muscle, indicating intramuscular adipogenesis is enhanced in Wagyu compared with Angus muscle.

  2. Mesenchymal and embryonic characteristics of stem cells obtained from mouse dental pulp. (United States)

    Guimarães, Elisalva Teixeira; Cruz, Gabriela Silva; de Jesus, Alan Araújo; Lacerda de Carvalho, Acácia Fernandes; Rogatto, Silvia Regina; Pereira, Lygia da Veiga; Ribeiro-dos-Santos, Ricardo; Soares, Milena Botelho Pereira


    Several studies have demonstrated that human dental pulp is a source of mesenchymal stem cells. To better understand the biological properties of these cells we isolated and characterized stem cells from the dental pulp of EGFP transgenic mice. The pulp tissue was gently separated from the roots of teeth extracted from C57BL/6 mice, and cultured under appropriate conditions. Flow cytometry, RT-PCR, light microscopy (staining for alkaline phosphatase) and immunofluorescence were used to investigate the expression of stem cell markers. The presence of chromosomal abnormalities was evaluated by G banding. The mouse dental pulp stem cells (mDPSC) were highly proliferative, plastic-adherent, and exhibited a polymorphic morphology predominantly with stellate or fusiform shapes. The presence of cell clusters was observed in cultures of mDPSC. Some cells were positive for alkaline phosphatase. The karyotype was normal until the 5th passage. The Pou5f1/Oct-4 and ZFP42/Rex-1, but not Nanog transcripts were detected in mDPSC. Flow cytometry and fluorescence analyses revealed the presence of a heterogeneous population positive for embryonic and mesenchymal cell markers. Adipogenic, chondrogenic and osteogenic differentiation was achieved after two weeks of cell culture under chemically defined in vitro conditions. In addition, some elongated cells spontaneously acquired a contraction capacity. Our results reinforce that the dental pulp is an important source of adult stem cells and encourage studies on therapeutic potential of mDPSC in experimental disease models. Copyright © 2011 Elsevier Ltd. All rights reserved.

  3. Hierarchical mechanisms for transcription factor-mediated reprogramming of fibroblasts to neurons (United States)

    Wapinski, Orly L.; Vierbuchen, Thomas; Qu, Kun; Lee, Qian Yi; Chanda, Soham; Fuentes, Daniel R.; Giresi, Paul G.; Ng, Yi Han; Marro, Samuele; Neff, Norma F.; Drechsel, Daniela; Martynoga, Ben; Castro, Diogo S.; Webb, Ashley E.; Brunet, Anne; Guillemot, Francois; Chang, Howard Y.; Wernig, Marius


    SUMMARY Direct lineage reprogramming is a promising approach for human disease modeling and regenerative medicine with poorly understood mechanisms. Here we reveal a hierarchical mechanism in the direct conversion of fibroblasts into induced neuronal (iN) cells mediated by the transcription factors Ascl1, Brn2, and Myt1l. Ascl1 acts as an “on target” pioneer factor by immediately occupying most cognate genomic sites in fibroblasts. In contrast, Brn2 and Myt1l do not access fibroblast chromatin productively on their own; instead Ascl1 recruits Brn2 to Ascl1 sites genome-wide. A unique trivalent chromatin signature in the host cells predicts the permissiveness for Ascl1 pioneering activity among different cell types. Finally, we identified Zfp238 as a key Ascl1 target gene that can partially substitute for Ascl1 during iN cell reprogramming. Thus, precise match between pioneer factor and the chromatin context at key target genes is determinative for trans-differentiation to neurons and likely other cell types. PMID:24243019

  4. DNA hypomethylation, transient neonatal diabetes, and prune belly sequence in one of two identical twins. (United States)

    Laborie, Lene Bjerke; Mackay, Deborah J G; Temple, I Karen; Molven, Anders; Søvik, Oddmund; Njølstad, Pål Rasmus


    One known genetic mechanism for transient neonatal diabetes is loss of methylation at 6q24. The etiology of prune belly sequence is unknown but a genetic defect, affecting the mesoderm from which the triad abdominal muscle hypoplasia, urinary tract abnormalities, and cryptorchidism develop, has been suggested. We investigated a family, including one twin, with transient neonatal diabetes and prune belly sequence. Autoantibody tests excluded type 1 diabetes. Microsatellite marker analysis confirmed the twins being monozygotic. We identified no mutations in ZFP57, KCNJ11, ABCC8, GCK, HNF1A, HNF1B, HNF3B, IPF1, PAX4, or ZIC3. The proband had loss of methylation at the 6q24 locus TNDM and also at the loci IGF2R, DIRAS3, and PEG1, while the other family members, including the healthy monozygotic twin, had normal findings. The loss of methylation on chromosome 6q24 and elsewhere may indicate a generalized maternal hypomethylation syndrome, which accounts for both transient neonatal diabetes and prune belly sequence.

  5. Single-Cell Landscape of Transcriptional Heterogeneity and Cell Fate Decisions during Mouse Early Gastrulation

    Directory of Open Access Journals (Sweden)

    Hisham Mohammed


    Full Text Available The mouse inner cell mass (ICM segregates into the epiblast and primitive endoderm (PrE lineages coincident with implantation of the embryo. The epiblast subsequently undergoes considerable expansion of cell numbers prior to gastrulation. To investigate underlying regulatory principles, we performed systematic single-cell RNA sequencing (seq of conceptuses from E3.5 to E6.5. The epiblast shows reactivation and subsequent inactivation of the X chromosome, with Zfp57 expression associated with reactivation and inactivation together with other candidate regulators. At E6.5, the transition from epiblast to primitive streak is linked with decreased expression of polycomb subunits, suggesting a key regulatory role. Notably, our analyses suggest elevated transcriptional noise at E3.5 and within the non-committed epiblast at E6.5, coinciding with exit from pluripotency. By contrast, E6.5 primitive streak cells became highly synchronized and exhibit a shortened G1 cell-cycle phase, consistent with accelerated proliferation. Our study systematically charts transcriptional noise and uncovers molecular processes associated with early lineage decisions.

  6. Zinc finger protein 521 antagonizes early B-cell factor 1 and modulates the B-lymphoid differentiation of primary hematopoietic progenitors. (United States)

    Mega, Tiziana; Lupia, Michela; Amodio, Nicola; Horton, Sarah J; Mesuraca, Maria; Pelaggi, Daniela; Agosti, Valter; Grieco, Michele; Chiarella, Emanuela; Spina, Raffaella; Moore, Malcolm A S; Schuringa, Jan Jacob; Bond, Heather M; Morrone, Giovanni


    Zinc finger protein 521 (EHZF/ZNF521) is a multi-functional transcription co-factor containing 30 zinc fingers and an amino-terminal motif that binds to the nucleosome remodelling and histone deacetylase (NuRD) complex. ZNF521 is believed to be a relevant player in the regulation of the homeostasis of the hematopoietic stem/progenitor cell compartment, however the underlying molecular mechanisms are still largely unknown. Here, we show that this protein plays an important role in the control of B-cell development by inhibiting the activity of early B-cell factor-1 (EBF1), a master factor in B-lineage specification. In particular, our data demonstrate that: (1) ZNF521 binds to EBF1 via its carboxyl-terminal portion and this interaction is required for EBF1 inhibition; (2) NuRD complex recruitment by ZNF521 is not essential for the inhibition of transactivation of EBF1-dependent promoters; (3) ZNF521 represses EBF1 target genes in a human B-lymphoid molecular context; and (4) RNAi-mediated silencing of ZNF521/Zfp521 in primary human and murine hematopoietic progenitors strongly enhances the generation of B-lymphocytes in vitro. Taken together, our data indicate that ZNF521 can antagonize B-cell development and lend support to the notion that it may contribute to conserve the multipotency of primitive lympho-myeloid progenitors by preventing or delaying their EBF1-driven commitment toward the B-cell lineage.

  7. Development of manufacturing process for production of 500 MWe calandria sheets

    International Nuclear Information System (INIS)

    Hariharan, R.; Ramesh, P.; Lakshminarayana, B.; Bhaskara Rao, C.V.; Pande, P.; Agarwala, G.C.


    Calandria tubes made of zircaloy-2 are being used as structural components in pressurised heavy water power reactors. The sheets required for producing calandria tube for 235 MWe reactors are being manufactured at Zircaloy Fabrication Plant (ZFP), NFC utilizing a 2 Hi/4 Hi rolling mill procured for the purpose, by carrying out cold rolling process to achieve the required size after hot rolling suitable extruded slabs. Due to limitation of width of the sheet that can be rolled with the mill as well as the size of the slab that can be extruded with the existing press, difficulties arose in producing acceptable full length sheets of size 6600 mm long x 435 mm wide x 1.6 mm thick for manufacturing 500 MWe calandria tube. This paper deals with the details of the process problem resolved. They are: (a)designing of suitable hot and cold rolling pass schedules, (b)selection and standardization of process parameters such as beta quenching, hot rolling and cold rolling, and (c)details of the overall manufacturing process. Due to implementation of above, sheets required for manufacturing 500 MWe calandria tube sheets were successfully rolled. About 40 nos. of acceptable full length sheets have already been manufactured. (author). 1 fig., 3 tabs

  8. Culture medium, gas atmosphere and MAPK inhibition affect regulation of RNA-binding protein targets during mouse preimplantation development. (United States)

    Calder, Michele D; Watson, Patricia H; Watson, Andrew J


    During oogenesis, mammalian oocytes accumulate maternal mRNAs that support the embryo until embryonic genome activation. RNA-binding proteins (RBP) may regulate the stability and turnover of maternal and embryonic mRNAs. We hypothesised that varying embryo culture conditions, such as culture medium, oxygen tension and MAPK inhibition, affects regulation of RBPs and their targets during preimplantation development. STAU1, ELAVL1, KHSRP and ZFP36 proteins and mRNAs were detected throughout mouse preimplantation development, whereas Elavl2 mRNA decreased after the two-cell stage. Potential target mRNAs of RBP regulation, Gclc, Slc2a1 and Slc7a1 were detected during mouse preimplantation development. Gclc mRNA was significantly elevated in embryos cultured in Whitten's medium compared with embryos cultured in KSOMaa, and Gclc mRNA was elevated under high-oxygen conditions. Inhibition of the p38 MAPK pathway reduced Slc7a1 mRNA expression while inhibition of ERK increased Slc2a1 mRNA expression. The half-lives of the potential RBP mRNA targets are not regulated in parallel; Slc2a1 mRNA displayed the longest half-life. Our results indicate that mRNAs and proteins encoding five RBPs are present during preimplantation development and more importantly, demonstrate that expression of RBP target mRNAs are regulated by culture medium, gas atmosphere and MAPK pathways.

  9. Histone deacetylase 3 is required for maintenance of bone mass during aging (United States)

    McGee-Lawrence, Meghan E.; Bradley, Elizabeth W.; Dudakovic, Amel; Carlson, Samuel W.; Ryan, Zachary C.; Kumar, Rajiv; Dadsetan, Mahrokh; Yaszemski, Michael J.; Chen, Qingshan; An, Kai-Nan; Westendorf, Jennifer J.


    Histone deacetylase 3 (Hdac3) is a nuclear enzyme that removes acetyl groups from lysine residues in histones and other proteins to epigenetically regulate gene expression. Hdac3 interacts with bone-related transcription factors and co-factors such as Runx2 and Zfp521, and thus is poised to play a key role in the skeletal system. To understand the role of Hdac3 in osteoblasts and osteocytes, Hdac3 conditional knockout (CKO) mice were created with the Osteocalcin (OCN) promoter driving Cre expression. Hdac3 CKOOCN mice were of normal size and weight, but progressively lost trabecular and cortical bone mass with age. The Hdac3 CKOOCN mice exhibited reduced cortical bone mineralization and material properties and suffered frequent fractures. Bone resorption was lower, not higher, in the Hdac3 CKOOCN mice, suggesting that primary defects in osteoblasts caused the reduced bone mass. Indeed, reductions in bone formation were observed. Osteoblasts and osteocytes from Hdac3 CKOOCN mice showed increased DNA damage and reduced functional activity in vivo and in vitro. Thus, Hdac3 expression in osteoblasts and osteocytes is essential for bone maintenance during aging. PMID:23085085

  10. Central Role of the PPARγ Gene Network in Coordinating Beef Cattle Intramuscular Adipogenesis in Response to Weaning Age and Nutrition. (United States)

    Moisá, Sonia J; Shike, Daniel W; Faulkner, Dan B; Meteer, William T; Keisler, Duane; Loor, Juan J


    Adipogenic/lipogenic transcriptional networks regulating intramuscular fat deposition (IMF) in response to weaning age and dietary starch level were studied. The longissimus muscle (LM) of beef steers on an early weaning (141 days age) plus high-starch diet (EWS) or a normal weaning (NW, 222 days age) plus starch creep-feed diet (CFS) was biopsied at 0 (EW), 25, 50, 96 (NW), 167, and 222 (pre-slaughter) days. Expression patterns of 35 target genes were studied. From NW through slaughter, all steers received the same high-starch diet. In EWS steers the expression of PPARG, other adipogenic (CEBPA, ZFP423) and lipogenic (THRSP, SREBF1, INSIG1) activators, and several enzymes (FASN, SCD, ELOVL6, PCK1, DGAT2) that participate in the process of IMF increased gradually to a peak between 96 and 167 days on treatment. Steers in NW did not achieve similar expression levels even by 222 days on treatment, suggesting a blunted response even when fed a high-starch diet after weaning. High-starch feeding at an early age (EWS) triggers precocious and sustained adipogenesis, resulting in greater marbling.

  11. Combined analysis of mRNA and miRNA identifies dehydration and salinity responsive key molecular players in citrus roots. (United States)

    Xie, Rangjin; Zhang, Jin; Ma, Yanyan; Pan, Xiaoting; Dong, Cuicui; Pang, Shaoping; He, Shaolan; Deng, Lie; Yi, Shilai; Zheng, Yongqiang; Lv, Qiang


    Citrus is one of the most economically important fruit crops around world. Drought and salinity stresses adversely affected its productivity and fruit quality. However, the genetic regulatory networks and signaling pathways involved in drought and salinity remain to be elucidated. With RNA-seq and sRNA-seq, an integrative analysis of miRNA and mRNA expression profiling and their regulatory networks were conducted using citrus roots subjected to dehydration and salt treatment. Differentially expressed (DE) mRNA and miRNA profiles were obtained according to fold change analysis and the relationships between miRNAs and target mRNAs were found to be coherent and incoherent in the regulatory networks. GO enrichment analysis revealed that some crucial biological processes related to signal transduction (e.g. 'MAPK cascade'), hormone-mediated signaling pathways (e.g. abscisic acid- activated signaling pathway'), reactive oxygen species (ROS) metabolic process (e.g. 'hydrogen peroxide catabolic process') and transcription factors (e.g., 'MYB, ZFP and bZIP') were involved in dehydration and/or salt treatment. The molecular players in response to dehydration and salt treatment were partially overlapping. Quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) analysis further confirmed the results from RNA-seq and sRNA-seq analysis. This study provides new insights into the molecular mechanisms how citrus roots respond to dehydration and salt treatment.

  12. Somatic drivers of B-ALL in a model of ETV6-RUNX1; Pax5+/− leukemia

    International Nuclear Information System (INIS)

    Weyden, Louise van der; Giotopoulos, George; Wong, Kim; Rust, Alistair G.; Robles-Espinoza, Carla Daniela; Osaki, Hikari; Huntly, Brian J.; Adams, David J.


    B-cell precursor acute lymphoblastic leukemia (B-ALL) is amongst the leading causes of childhood cancer-related mortality. Its most common chromosomal aberration is the ETV6-RUNX1 fusion gene, with ~25 % of ETV6-RUNX1 patients also carrying PAX5 alterations. We have recreated this mutation background by inter-crossing Etv6-RUNX1 (Etv6 RUNX1-SB ) and Pax5 +/− mice and performed an in vivo analysis to find driver genes using Sleeping Beauty transposon-mediated mutagenesis and also exome sequencing. Combination of Etv6-RUNX1 and Pax5 +/− alleles generated a transplantable B220 + CD19+ B-ALL with a significant disease incidence. RNA-seq analysis showed a gene expression pattern consistent with arrest at the pre-B stage. Analysis of the transposon common insertion sites identified genes involved in B-cell development (Zfp423) and the JAK/STAT signaling pathway (Jak1, Stat5 and Il2rb), while exome sequencing revealed somatic hotspot mutations in Jak1 and Jak3 at residues analogous to those mutated in human leukemias, and also mutation of Trp53. Powerful synergies exists in our model suggesting STAT pathway activation and mutation of Trp53 are potent drivers of B-ALL in the context of Etv6-RUNX1;Pax5 +/− . The online version of this article (doi:10.1186/s12885-015-1586-1) contains supplementary material, which is available to authorized users

  13. Recent advances in lossy compression of scientific floating-point data (United States)

    Lindstrom, P.


    With a continuing exponential trend in supercomputer performance, ever larger data sets are being generated through numerical simulation. Bandwidth and storage capacity are, however, not keeping pace with this increase in data size, causing significant data movement bottlenecks in simulation codes and substantial monetary costs associated with archiving vast volumes of data. Worse yet, ever smaller fractions of data generated can be stored for further analysis, where scientists frequently rely on decimating or averaging large data sets in time and/or space. One way to mitigate these problems is to employ data compression to reduce data volumes. However, lossless compression of floating-point data can achieve only very modest size reductions on the order of 10-50%. We present ZFP and FPZIP, two state-of-the-art lossy compressors for structured floating-point data that routinely achieve one to two orders of magnitude reduction with little to no impact on the accuracy of visualization and quantitative data analysis. We provide examples of the use of such lossy compressors in climate and seismic modeling applications to effectively accelerate I/O and reduce storage requirements. We further discuss how the design decisions behind these and other compressors impact error distributions and other statistical and differential properties, including derived quantities of interest relevant to each science application.

  14. Computational modeling identifies key gene regulatory interactions underlying phenobarbital-mediated tumor promotion (United States)

    Luisier, Raphaëlle; Unterberger, Elif B.; Goodman, Jay I.; Schwarz, Michael; Moggs, Jonathan; Terranova, Rémi; van Nimwegen, Erik


    Gene regulatory interactions underlying the early stages of non-genotoxic carcinogenesis are poorly understood. Here, we have identified key candidate regulators of phenobarbital (PB)-mediated mouse liver tumorigenesis, a well-characterized model of non-genotoxic carcinogenesis, by applying a new computational modeling approach to a comprehensive collection of in vivo gene expression studies. We have combined our previously developed motif activity response analysis (MARA), which models gene expression patterns in terms of computationally predicted transcription factor binding sites with singular value decomposition (SVD) of the inferred motif activities, to disentangle the roles that different transcriptional regulators play in specific biological pathways of tumor promotion. Furthermore, transgenic mouse models enabled us to identify which of these regulatory activities was downstream of constitutive androstane receptor and β-catenin signaling, both crucial components of PB-mediated liver tumorigenesis. We propose novel roles for E2F and ZFP161 in PB-mediated hepatocyte proliferation and suggest that PB-mediated suppression of ESR1 activity contributes to the development of a tumor-prone environment. Our study shows that combining MARA with SVD allows for automated identification of independent transcription regulatory programs within a complex in vivo tissue environment and provides novel mechanistic insights into PB-mediated hepatocarcinogenesis. PMID:24464994

  15. [Psychiatric care between World War II and "Psychiatrie-Enquête": early approaches of reformation in Württemberg]. (United States)

    Croissant, Daniela; Längle, Gerhard


    The article describes care in a psychiatric clinic between 1946 and 1975. This happens against the background of the current psychiatry-historical literature in which this phase of psychiatric care is described often summarily with the destructive words of the report of the 'Psychiatrie-Enquête' of 1975. Improvements achieved in this time were hardly examined up to now though they contributed substantially to the later effects of the 'Psychiatrie-Enquête'. The medical annual reports of the psychiatric clinic of Zwiefalten, today ZfP Südwürttemberg, refering to the mentioned period were sighted and evaluated concerning their contents. In the called period evident organizational and structural defects are deplored in the annual reports. Nevertheless, from the late 1940 s on, modern care elements appear, as for example the broadening of the range of the therapeutic offers, multiprofessional treatment, diagnosis-specific concepts for the wards, opening of stations and extensive outpatient care. It is shown that already before the appearance of the final report of the Enquête commission clear progress concerning psychiatric care was achieved. © Georg Thieme Verlag KG Stuttgart · New York.

  16. Identification of Yellow Pigmentation Genes in Brassica rapa ssp. pekinensis Using Br300 Microarray

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    Hee-Jeong Jung


    Full Text Available The yellow color of inner leaves in Chinese cabbage depends on its lutein and carotene content. To identify responsible genes for yellow pigmentation in leaves, the transcriptome profiles of white (Kenshin and yellow leaves (Wheessen were examined using the Br300K oligomeric chip in Chinese cabbage. In yellow leaves, genes involved in carotene synthesis (BrPSY, BrPDS, BrCRTISO, and BrLCYE, lutein, and zeaxanthin synthesis (BrCYP97A3 and BrHYDB were upregulated, while those associated with carotene degradation (BrNCED3, BrNCED4, and BrNCED6 were downregulated. These expression patterns might support that the content of both lutein and total carotenoid was much higher in the yellow leaves than that in the white leaves. These results indicate that the yellow leaves accumulate high levels of both lutein and β-carotene due to stimulation of synthesis and that the degradation rate is inhibited. A large number of responsible genes as novel genes were specifically expressed in yellow inner leaves, suggesting the possible involvement in pigment synthesis. Finally, we identified three transcription factors (BrA20/AN1-like, BrBIM1, and BrZFP8 that are specifically expressed and confirmed their relatedness in carotenoid synthesis from Arabidopsis plants.

  17. Intelligent robot / manipulator systems for NDT of primary components in nuclear power plants; Intelligente Roboter / Handhabungssysteme fuer die Pruefung von Primaerkreiskomponenten in Kernkraftwerken

    Energy Technology Data Exchange (ETDEWEB)

    Dirauf, F.; Gottfried, R.; Bauer, R. [Siemens AG, KWU, Erlangen (Germany)


    Gruppenstrahlenpruefkoepfen entwickelt. Die zerstoerungsfreie Pruefung von Stutzen im Reaktordruckbehaelter von Siedewasserreaktoren erfolgte ueblicherweise von der Aussenseite der Komponente. Der von Siemens neu entwickelte Stutzenmanipulator kann mittels Handhabungsgestaenge oder optional durch Schwimmeinrichtung an den zu pruefenden Stutzen gefahren und dort durch pneumatische Saugteller fixiert werden. Aufgrund des modularen Aufbaus koennen unterschiedliche Pruefkopfsysteme fuer die verschiedenen Stutzengroessen und Pruefbereiche, wie z.B. Stutzenkanten, Stutzeninnenbereiche und Stutzeneinschweissnaehte, montiert werden. Der mobile Pruefroboter SISTAR fuer die Pruefung von DWR-Reaktordruckbehaeltern besteht aus einer schwimmfaehigen zylinderfoermigen Plattform, die sich unter Wasser mit Propellerantrieben oder mit Unterstuetzung durch Halteseile in ihre vorgesehene Position im Reaktordruckbehaelter bewegt. Dort richtet sie sich selbsttaetig horizontal aus und fixiert sich im Zentrum des RDB mit Hilfe von Beinen die synchron radial ausgefahren werden. Je nach Pruefaufgabe wird die Plattform mit einem oder zwei Roboterarmen ausgeruestet. (orig./DGE)

  18. The Promyelocytic Leukemia Zinc Finger Protein: Two Decades of Molecular Oncology

    International Nuclear Information System (INIS)

    Suliman, Bandar Ali; Xu, Dakang; Williams, Bryan Raymond George


    The promyelocytic leukemia zinc finger (PLZF) protein, also known as Zbtb16 or Zfp145, was first identified in a patient with acute promyelocytic leukemia, where a reciprocal chromosomal translocation t(11;17)(q23;q21) resulted in a fusion with the RARA gene encoding retinoic acid receptor alpha. The wild-type Zbtb16 gene encodes a transcription factor that belongs to the POK (POZ and Krüppel) family of transcriptional repressors. In addition to nine Krüppel-type sequence-specific zinc fingers, which make it a member of the Krüppel-like zinc finger protein family, the PLZF protein contains an N-terminal BTB/POZ domain and RD2 domain. PLZF has been shown to be involved in major developmental and biological processes, such as spermatogenesis, hind limb formation, hematopoiesis, and immune regulation. PLZF is localized mainly in the nucleus where it exerts its transcriptional repression function, and many post-translational modifications affect this ability and also have an impact on its cytoplasmic/nuclear dissociation. PLZF achieves its transcriptional regulation by binding to many secondary molecules to form large multi-protein complexes that bind to the regulatory elements in the promoter region of the target genes. These complexes are also capable of physically interacting with its target proteins. Recently, PLZF has become implicated in carcinogenesis as a tumor suppressor gene, since it regulates the cell cycle and apoptosis in many cell types. This review will examine the major advances in our knowledge of PLZF biological activities that augment its value as a therapeutic target, particularly in cancer and immunological diseases.

  19. Molecular Factors Underlying the Deposition of Intramuscular Fat and Collagen in Skeletal Muscle of Nellore and Angus Cattle. (United States)

    Martins, Taiane S; Sanglard, Letícia M P; Silva, Walmir; Chizzotti, Mário L; Rennó, Luciana N; Serão, Nick V L; Silva, Fabyano F; Guimarães, Simone E F; Ladeira, Márcio M; Dodson, Michael V; Du, Min; Duarte, Marcio S


    Studies have shown that intramuscular adipogenesis and fibrogenesis may concomitantly occur in skeletal muscle of beef cattle. Thus, we hypothesized that the discrepancy of intramuscular fat content in beef from Nellore and Angus was associated with differences in intramuscular adipogenesis and fibrogenesis during the finishing phase. To test our hypothesis, longissimus muscle samples of Nellore (n = 6; BW = 372.5 ± 37.3 kg) and Angus (n = 6; BW = 382.8 ± 23.9 kg) cattle were collected for analysis of gene and protein expression, and quantification of intramuscular fat and collagen. Least-squares means were estimated for the effect of Breed and differences were considered at P ≤ 0.05. A greater intramuscular fat content was observed in skeletal muscle of Angus compared to Nellore cattle (P≤0.05). No differences were observed for mRNA expression of lipogenic and lipolytic markers ACC, FAS, FABP4, SERBP-1, CPT-2, LPL, and ACOX (P > 0.05) in skeletal muscle of Nellore and Angus cattle. Similarly, no differences were observed in mRNA expression of adipogenic markers Zfp423, PPARγ, and C/EBPα (P>0.05) However, a greater PPARγ protein content was observed in skeletal muscle of Angus compared to Nellore cattle (P≤0.05). A greater abundance of adipo/fibrogenic cells, evaluated by the PDGFRα content, was observed in skeletal muscle of Angus than Nellore cattle (P≤0.05). No differences in fibrogenesis were observed in skeletal muscle of Angus and Nellore cattle, which is in accordance with the lack of differences in intramuscular collagen content in beef from both breeds (P>0.05). These findings demonstrate that difference in intramuscular fat content is associated with a slightly enhanced adipogenesis in skeletal muscle of Angus compared to Nellore cattle, while no difference in fibrogenesis.

  20. Methylomic changes in individuals with psychosis, prenatally exposed to endocrine disrupting compounds: Lessons from diethylstilbestrol.

    Directory of Open Access Journals (Sweden)

    Fabrice Rivollier

    Full Text Available In the Western world, between 1940 and 1970, more than 2 million people were exposed in utero to diethylstilbestrol (DES. In exposed individuals, and in their descendants, adverse outcomes have been linked to such exposure, including cancers, genital malformations, and less consistently, psychiatric disorders. We aimed to explore whether prenatal DES exposure would be associated with DNA methylation changes, and whether these epigenetic modifications would be associated with increased risk of psychosis.From 247 individuals born from mothers exposed to DES, we selected 69 siblings from 30 families. In each family, at least one sibling was exposed in utero to DES. We performed a methylome-wide association study using HumanMethylation450 DNA Analysis BeadChip® in peripheral blood. We analyzed methylation changes at individual CpGs or regions in exposed (n = 37 versus unexposed individuals (n = 32. We also compared exposed individuals with (n = 7 and without psychosis (n = 30.There were more individuals with schizophrenia in the DES-exposed group. We found no significant differences between exposed and unexposed individuals with respect to differentially methylated CpGs or regions. The largest difference was in a region near the promoter of an ADAMTS proteoglycanase gene (ADAMTS9. Compared to exposed individuals without psychosis, exposed individuals with psychosis had differential methylation in the region encompassing the gene encoding the zinc finger protein 57 (ZFP57.In utero exposure to DES was not associated with methylation changes at specific CpG or regions. In exposed individuals, however, psychosis was associated with specific methylomic modifications that could impact neurodevelopment and neuroplasticity.

  1. SOX9 as a Predictor for Neurogenesis Potentiality of Amniotic Fluid Stem Cells (United States)

    Wei, Pei-Cih; Chao, Angel; Peng, Hsiu-Huei; Chao, An-Shine; Chang, Yao-Lung; Chang, Shuenn-Dyh; Wang, Hsin-Shih; Chang, Yu-Jen; Tsai, Ming-Song; Sieber, Martin; Chen, Hua-Chien; Chen, Shu-Jen; Lee, Yun-Shien


    Preclinical studies of amniotic fluid-derived cell therapy have been successful in the research of neurodegenerative diseases, peripheral nerve injury, spinal cord injury, and brain ischemia. Transplantation of human amniotic fluid stem cells (AFSCs) into rat brain ventricles has shown improvement in symptoms of Parkinson's disease and also highlighted the minimal immune rejection risk of AFSCs, even between species. Although AFSCs appeared to be a promising resource for cell-based regenerative therapy, AFSCs contain a heterogeneous pool of distinct cell types, rendering each preparation of AFSCs unique. Identification of predictive markers for neuron-prone AFSCs is necessary before such stem cell-based therapeutics can become a reality. In an attempt to identify markers of AFSCs to predict their ability for neurogenesis, we performed a two-phase study. In the discovery phase of 23 AFSCs, we tested ZNF521/Zfp521, OCT6, SOX1, SOX2, SOX3, and SOX9 as predictive markers of AFSCs for neural differentiation. In the validation phase, the efficacy of these predictive markers was tested in independent sets of 18 AFSCs and 14 dental pulp stem cells (DPSCs). We found that high expression of SOX9 in AFSCs is associated with good neurogenetic ability, and these positive correlations were confirmed in independent sets of AFSCs and DPSCs. Furthermore, knockdown of SOX9 in AFSCs inhibited their neuronal differentiation. In conclusion, the discovery of SOX9 as a predictive marker for neuron-prone AFSCs could expedite the selection of useful clones for regenerative medicine, in particular, in neurological diseases and injuries. PMID:25154783

  2. Drought response in wheat: key genes and regulatory mechanisms controlling root system architecture and transpiration efficiency (United States)

    Kulkarni, Manoj; Soolanayakanahally, Raju; Ogawa, Satoshi; Uga, Yusaku; Selvaraj, Michael G.; Kagale, Sateesh


    Abiotic stresses such as drought, heat, salinity and flooding threaten global food security. Crop genetic improvement with increased resilience to abiotic stresses is a critical component of crop breeding strategies. Wheat is an important cereal crop and a staple food source globally. Enhanced drought tolerance in wheat is critical for sustainable food production and global food security. Recent advances in drought tolerance research have uncovered many key genes and transcription regulators governing morpho-physiological traits. Genes controlling root architecture and stomatal development play an important role in soil moisture extraction and its retention, and therefore have been targets of molecular breeding strategies for improving drought tolerance. In this systematic review, we have summarized evidence of beneficial contributions of root and stomatal traits to plant adaptation to drought stress. Specifically, we discuss a few key genes such as DRO1 in rice and ERECTA in Arabidopsis and rice that were identified to be the enhancers of drought tolerance via regulation of root traits and transpiration efficiency. Additionally, we highlight several transcription factor families, such as ERF (ethylene response factors), DREB (dehydration responsive element binding), ZFP (zinc finger proteins), WRKY and MYB that were identified to be both positive and negative regulators of drought responses in wheat, rice, maize and/or Arabidopsis. The overall aim of this review was to provide an overview of candidate genes that have been tested as regulators of drought response in plants. The lack of a reference genome sequence for wheat and nontransgenic approaches for manipulation of gene functions in the past had impeded high-resolution interrogation of functional elements, including genes and QTLs, and their application in cultivar improvement. The recent developments in wheat genomics and reverse genetics, including the availability of a gold-standard reference genome

  3. An eleven gene molecular signature for extra-capsular spread in oral squamous cell carcinoma serves as a prognosticator of outcome in patients without nodal metastases. (United States)

    Wang, Weining; Lim, Weng Khong; Leong, Hui Sun; Chong, Fui Teen; Lim, Tony K H; Tan, Daniel S W; Teh, Bin Tean; Iyer, N Gopalakrishna


    Extracapsular spread (ECS) is an important prognostic factor for oral squamous cell carcinoma (OSCC) and is used to guide management. In this study, we aimed to identify an expression profile signature for ECS in node-positive OSCC using data derived from two different sources: a cohort of OSCC patients from our institution (National Cancer Centre Singapore) and The Cancer Genome Atlas (TCGA) head and neck squamous cell carcinoma (HNSCC) cohort. We also sought to determine if this signature could serve as a prognostic factor in node negative cancers. Patients with a histological diagnosis of OSCC were identified from an institutional database and fresh tumor samples were retrieved. RNA was extracted and gene expression profiling was performed using the Affymetrix GeneChip Human Genome U133 Plus 2.0 microarray platform. RNA sequence data and corresponding clinical data for the TCGA HNSCC cohort were downloaded from the TCGA Data Portal. All data analyses were conducted using R package and SPSS. We identified an 11 gene signature (GGH, MTFR1, CDKN3, PSRC1, SMIM3, CA9, IRX4, CPA3, ZSCAN16, CBX7 and ZFP3) which was robust in segregating tumors by ECS status. In node negative patients, patients harboring this ECS signature had a significantly worse overall survival (p=0.04). An eleven gene signature for ECS was derived. Our results also suggest that this signature is prognostic in a separate subset of patients with no nodal metastasis Further validation of this signature on other datasets and immunohistochemical studies are required to establish utility of this signature in stratifying early stage OSCC patients. Copyright © 2014 Elsevier Ltd. All rights reserved.

  4. Inducing indel mutation in the SOX6 gene by zinc finger nuclease for gamma reactivation: An approach towards gene therapy of beta thalassemia. (United States)

    Modares Sadeghi, Mehran; Shariati, Laleh; Hejazi, Zahra; Shahbazi, Mansoureh; Tabatabaiefar, Mohammad Amin; Khanahmad, Hossein


    β-thalassemia is a common autosomal recessive disorder characterized by a deficiency in the synthesis of β-chains. Evidences show that increased HbF levels improve the symptoms in patients with β-thalassemia or sickle cell anemia. In this study, ZFN technology was applied to induce a mutation in the binding domain region of SOX6 to reactivate γ-globin expression. The sequences coding for ZFP arrays were designed and sub cloned in TDH plus as a transfer vector. The ZFN expression was confirmed using Western blot analysis. In the next step, using the site-directed mutagenesis strategy through the overlap PCR, a missense mutation (D64V) was induced in the catalytic domain of the integrase gene in the packaging plasmid and verified using DNA sequencing. Then, the integrase minus lentivirus containing ZFN cassette was packaged. Transduction of K562 cells with this virus was performed. Mutation detection assay was performed. The indel percentage of the cells transducted with lenti virus containing ZFN was 31%. After 5 days of erythroid differentiation with 15 μg/mL cisplatin, the levels of γ-globin mRNA were sixfold in the cells treated with ZFN compared to untreated cells. In the meantime, the measurement of HbF expression levels was carried out using hemoglobin electrophoresis and showed the same results. Integrase minus lentivirus can provide a useful tool for efficient transient gene expression and helps avoid disadvantages of gene targeting using the native virus. The ZFN strategy applied here to induce indel on SOX6 gene in adult erythroid progenitors may provide a method to activate fetal hemoglobin expression in individuals with β-thalassemia. © 2017 Wiley Periodicals, Inc.

  5. Profiling trait anxiety: transcriptome analysis reveals cathepsin B (Ctsb as a novel candidate gene for emotionality in mice.

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    Ludwig Czibere

    Full Text Available Behavioral endophenotypes are determined by a multitude of counteracting but precisely balanced molecular and physiological mechanisms. In this study, we aim to identify potential novel molecular targets that contribute to the multigenic trait "anxiety". We used microarrays to investigate the gene expression profiles of different brain regions within the limbic system of mice which were selectively bred for either high (HAB or low (LAB anxiety-related behavior, and also show signs of comorbid depression-like behavior. We identified and confirmed sex-independent differences in the basal expression of 13 candidate genes, using tissue from the entire brain, including coronin 7 (Coro7, cathepsin B (Ctsb, muscleblind-like 1 (Mbnl1, metallothionein 1 (Mt1, solute carrier family 25 member 17 (Slc25a17, tribbles homolog 2 (Trib2, zinc finger protein 672 (Zfp672, syntaxin 3 (Stx3, ATP-binding cassette, sub-family A member 2 (Abca2, ectonucleotide pyrophosphatase/phosphodiesterase 5 (Enpp5, high mobility group nucleosomal binding domain 3 (Hmgn3 and pyruvate dehydrogenase beta (Pdhb. Additionally, we confirmed brain region-specific differences in the expression of synaptotagmin 4 (Syt4.Our identification of about 90 polymorphisms in Ctsb suggested that this gene might play a critical role in shaping our mouse model's behavioral endophenotypes. Indeed, the assessment of anxiety-related and depression-like behaviors of Ctsb knock-out mice revealed an increase in depression-like behavior in females. Altogether, our results suggest that Ctsb has significant effects on emotionality, irrespective of the tested mouse strain, making it a promising target for future pharmacotherapy.

  6. Pluripotent State Induction in Mouse Embryonic Fibroblast Using mRNAs of Reprogramming Factors

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    Ahmed Kamel El-Sayed


    Full Text Available Reprogramming of somatic cells has great potential to provide therapeutic treatments for a number of diseases as well as provide insight into mechanisms underlying early embryonic development. Improvement of induced Pluripotent Stem Cells (iPSCs generation through mRNA-based methods is currently an area of intense research. This approach provides a number of advantages over previously used methods such as DNA integration and insertional mutagenesis. Using transfection of specifically synthesized mRNAs of various pluripotency factors, we generated iPSCs from mouse embryonic fibroblast (MEF cells. The genetic, epigenetic and functional properties of the iPSCs were evaluated at different times during the reprogramming process. We successfully introduced synthesized mRNAs, which localized correctly inside the cells and exhibited efficient and stable translation into proteins. Our work demonstrated a robust up-regulation and a gradual promoter de-methylation of the pluripotency markers, including non-transfected factors such as Nanog, SSEA-1 (stage-specific embryonic antigen 1 and Rex-1 (ZFP-42, zinc finger protein 42. Using embryonic stem cells (ESCs conditions to culture the iPS cells resulted in formation of ES-like colonies after approximately 12 days with only five daily repeated transfections. The colonies were positive for alkaline phosphatase and pluripotency-specific markers associated with ESCs. This study revealed the ability of pluripotency induction and generation of mouse mRNA induced pluripotent stem cells (mRNA iPSCs using transfection of specifically synthesized mRNAs of various pluripotency factors into mouse embryonic fibroblast (MEF cells. These generated iPSCs exhibited molecular and functional properties similar to ESCs, which indicate that this method is an efficient and viable alternative to ESCs and can be used for further biological, developmental and therapeutic investigations.

  7. Critical role of zinc finger protein 521 in the control of growth, clonogenicity and tumorigenic potential of medulloblastoma cells. (United States)

    Spina, Raffaella; Filocamo, Gessica; Iaccino, Enrico; Scicchitano, Stefania; Lupia, Michela; Chiarella, Emanuela; Mega, Tiziana; Bernaudo, Francesca; Pelaggi, Daniela; Mesuraca, Maria; Pazzaglia, Simonetta; Semenkow, Samantha; Bar, Eli E; Kool, Marcel; Pfister, Stefan; Bond, Heather M; Eberhart, Charles G; Steinkühler, Christian; Morrone, Giovanni


    The stem cell-associated transcription co-factor ZNF521 has been implicated in the control of hematopoietic, osteo-adipogenic and neural progenitor cells. ZNF521 is highly expressed in cerebellum and in particular in the neonatal external granule layer that contains candidate medulloblastoma cells-of-origin, and in the majority of human medulloblastomas. Here we have explored its involvement in the control of human and murine medulloblastoma cells. The effect of ZNF521 on growth and tumorigenic potential of human medulloblastoma cell lines as well as primary Ptc1-/+ mouse medulloblastoma cells was investigated in a variety of in vitro and in vivo assays, by modulating its expression using lentiviral vectors carrying the ZNF521 cDNA, or shRNAs that silence its expression. Enforced overexpression of ZNF521 in DAOY medulloblastoma cells significantly increased their proliferation, growth as spheroids and ability to generate clones in single-cell cultures and semisolid media, and enhanced their migratory ability in wound-healing assays. Importantly, ZNF521-expressing cells displayed a greatly enhanced tumorigenic potential in nude mice. All these activities required the ZNF521 N-terminal motif that recruits the nucleosome remodeling and histone deacetylase complex, which might therefore represent an appealing therapeutic target. Conversely, silencing of ZNF521 in human UW228 medulloblastoma cells that display high baseline expression decreased their proliferation, clonogenicity, sphere formation and wound-healing ability. Similarly, Zfp521 silencing in mouse Ptc1-/+ medulloblastoma cells drastically reduced their growth and tumorigenic potential. Our data strongly support the notion that ZNF521, through the recruitment of the NuRD complex, contributes to the clonogenic growth, migration and tumorigenicity of medulloblastoma cells.

  8. Identification of Genes Whose Expression Profile Is Associated with Non-Progression towards AIDS Using eQTLs (United States)

    Le Clerc, Sigrid; van Manen, Daniëlle; Coulonges, Cédric; Ulveling, Damien; Laville, Vincent; Labib, Taoufik; Taing, Lieng; Delaneau, Olivier; Montes, Matthieu; Schuitemaker, Hanneke; Zagury, Jean-François


    Background Many genome-wide association studies have been performed on progression towards the acquired immune deficiency syndrome (AIDS) and they mainly identified associations within the HLA loci. In this study, we demonstrate that the integration of biological information, namely gene expression data, can enhance the sensitivity of genetic studies to unravel new genetic associations relevant to AIDS. Methods We collated the biological information compiled from three databases of expression quantitative trait loci (eQTLs) involved in cells of the immune system. We derived a list of single nucleotide polymorphisms (SNPs) that are functional in that they correlate with differential expression of genes in at least two of the databases. We tested the association of those SNPs with AIDS progression in two cohorts, GRIV and ACS. Tests on permuted phenotypes of the GRIV and ACS cohorts or on randomised sets of equivalent SNPs allowed us to assess the statistical robustness of this method and to estimate the true positive rate. Results Eight genes were identified with high confidence (p = 0.001, rate of true positives 75%). Some of those genes had previously been linked with HIV infection. Notably, ENTPD4 belongs to the same family as CD39, whose expression has already been associated with AIDS progression; while DNAJB12 is part of the HSP90 pathway, which is involved in the control of HIV latency. Our study also drew our attention to lesser-known functions such as mitochondrial ribosomal proteins and a zinc finger protein, ZFP57, which could be central to the effectiveness of HIV infection. Interestingly, for six out of those eight genes, down-regulation is associated with non-progression, which makes them appealing targets to develop drugs against HIV. PMID:26367535

  9. Identification of Genes Whose Expression Profile Is Associated with Non-Progression towards AIDS Using eQTLs.

    Directory of Open Access Journals (Sweden)

    Jean-Louis Spadoni

    Full Text Available Many genome-wide association studies have been performed on progression towards the acquired immune deficiency syndrome (AIDS and they mainly identified associations within the HLA loci. In this study, we demonstrate that the integration of biological information, namely gene expression data, can enhance the sensitivity of genetic studies to unravel new genetic associations relevant to AIDS.We collated the biological information compiled from three databases of expression quantitative trait loci (eQTLs involved in cells of the immune system. We derived a list of single nucleotide polymorphisms (SNPs that are functional in that they correlate with differential expression of genes in at least two of the databases. We tested the association of those SNPs with AIDS progression in two cohorts, GRIV and ACS. Tests on permuted phenotypes of the GRIV and ACS cohorts or on randomised sets of equivalent SNPs allowed us to assess the statistical robustness of this method and to estimate the true positive rate.Eight genes were identified with high confidence (p = 0.001, rate of true positives 75%. Some of those genes had previously been linked with HIV infection. Notably, ENTPD4 belongs to the same family as CD39, whose expression has already been associated with AIDS progression; while DNAJB12 is part of the HSP90 pathway, which is involved in the control of HIV latency. Our study also drew our attention to lesser-known functions such as mitochondrial ribosomal proteins and a zinc finger protein, ZFP57, which could be central to the effectiveness of HIV infection. Interestingly, for six out of those eight genes, down-regulation is associated with non-progression, which makes them appealing targets to develop drugs against HIV.

  10. Deletions of the long arm of chromosome 5 define subgroups of T-cell acute lymphoblastic leukemia. (United States)

    La Starza, Roberta; Barba, Gianluca; Demeyer, Sofie; Pierini, Valentina; Di Giacomo, Danika; Gianfelici, Valentina; Schwab, Claire; Matteucci, Caterina; Vicente, Carmen; Cools, Jan; Messina, Monica; Crescenzi, Barbara; Chiaretti, Sabina; Foà, Robin; Basso, Giuseppe; Harrison, Christine J; Mecucci, Cristina


    Recurrent deletions of the long arm of chromosome 5 were detected in 23/200 cases of T-cell acute lymphoblastic leukemia. Genomic studies identified two types of deletions: interstitial and terminal. Interstitial 5q deletions, found in five cases, were present in both adults and children with a female predominance (chi-square, P=0.012). Interestingly, these cases resembled immature/early T-cell precursor acute lymphoblastic leukemia showing significant down-regulation of five out of the ten top differentially expressed genes in this leukemia group, including TCF7 which maps within the 5q31 common deleted region. Mutations of genes known to be associated with immature/early T-cell precursor acute lymphoblastic leukemia, i.e. WT1, ETV6, JAK1, JAK3, and RUNX1, were present, while CDKN2A/B deletions/mutations were never detected. All patients had relapsed/resistant disease and blasts showed an early differentiation arrest with expression of myeloid markers. Terminal 5q deletions, found in 18 of patients, were more prevalent in adults (chi-square, P=0.010) and defined a subgroup of HOXA-positive T-cell acute lymphoblastic leukemia characterized by 130 up- and 197 down-regulated genes. Down-regulated genes included TRIM41, ZFP62, MAPK9, MGAT1, and CNOT6, all mapping within the 1.4 Mb common deleted region at 5q35.3. Of interest, besides CNOT6 down-regulation, these cases also showed low BTG1 expression and a high incidence of CNOT3 mutations, suggesting that the CCR4-NOT complex plays a crucial role in the pathogenesis of HOXA-positive T-cell acute lymphoblastic leukemia with terminal 5q deletions. In conclusion, interstitial and terminal 5q deletions are recurrent genomic losses identifying distinct subtypes of T-cell acute lymphoblastic leukemia. Copyright© Ferrata Storti Foundation.

  11. The insulin/IGF signaling regulators cytohesin/GRP-1 and PIP5K/PPK-1 modulate susceptibility to excitotoxicity in C. elegans.

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    Nazila Tehrani

    Full Text Available During ischemic stroke, malfunction of excitatory amino acid transporters and reduced synaptic clearance causes accumulation of Glutamate (Glu and excessive stimulation of postsynaptic neurons, which can lead to their degeneration by excitotoxicity. The balance between cell death-promoting (neurotoxic and survival-promoting (neuroprotective signaling cascades determines the fate of neurons exposed to the excitotoxic insult. The evolutionary conserved Insulin/IGF Signaling (IIS cascade can participate in this balance, as it controls cell stress resistance in nematodes and mammals. Blocking the IIS cascade allows the transcription factor FoxO3/DAF-16 to accumulate in the nucleus and activate a transcriptional program that protects cells from a range of insults. We study the effect of IIS cascade on neurodegeneration in a C. elegans model of excitotoxicity, where a mutation in a central Glu transporter (glt-3 in a sensitizing background causes Glu-Receptor -dependent neuronal necrosis. We expand our studies on the role of the IIS cascade in determining susceptibility to excitotoxic necrosis by either blocking IIS at the level of PI3K/AGE-1 or stimulating it by removing the inhibitory effect of ZFP-1 on the expression of PDK-1. We further show that the components of the Cytohesin/GRP-1, Arf, and PIP5K/PPK-1 complex, known to regulate PIP2 production and the IIS cascade, modulate nematode excitotoxicity: mutations that are expected to reduce the complex's ability to produce PIP2 and inhibit the IIS cascade protect from excitotoxicity, while overstimulation of PIP2 production enhances neurodegeneration. Our observations therefore affirm the importance of the IIS cascade in determining the susceptibility to necrotic neurodegeneration in nematode excitotoxicity, and demonstrate the ability of Cytohesin/GRP-1, Arf, and PIP5K/PPK-1 complex to modulate neuroprotection.

  12. The stabilization of hypoxia inducible factor modulates differentiation status and inhibits the proliferation of mouse embryonic stem cells. (United States)

    Binó, Lucia; Kučera, Jan; Štefková, Kateřina; Švihálková Šindlerová, Lenka; Lánová, Martina; Kudová, Jana; Kubala, Lukáš; Pacherník, Jiří


    Hypoxic conditions are suggested to affect the differentiation status of stem cells (SC), including embryonic stem cells (ESC). Hypoxia inducible factor (HIF) is one of the main intracellular molecules responsible for the cellular response to hypoxia. Hypoxia stabilizes HIF by inhibiting the activity of HIF prolyl-hydroxylases (PHD), which are responsible for targeting HIF-alpha subunits for proteosomal degradation. To address the impact of HIF stabilization on the maintenance of the stemness signature of mouse ESC (mESC), we tested the influence of the inhibition of PHDs and hypoxia (1% O2 and 5% O2) on spontaneous ESC differentiation triggered by leukemia inhibitory factor withdrawal for 24 and 48 h. The widely used panhydroxylase inhibitor dimethyloxaloylglycine (DMOG) and PHD inhibitor JNJ-42041935 (JNJ) with suggested higher specificity towards PHDs were employed. Both inhibitors and both levels of hypoxia significantly increased HIF-1alpha and HIF-2alpha protein levels and HIF transcriptional activity in spontaneously differentiating mESC. This was accompanied by significant downregulation of cell proliferation manifested by the complete inhibition of DNA synthesis and partial arrest in the S phase after 48 h. Further, HIF stabilization enhanced downregulation of the expressions of some pluripotency markers (OCT-4, NANOG, ZFP-42, TNAP) in spontaneously differentiating mESC. However, at the same time, there was also a significant decrease in the expression of some genes selected as markers of cell differentiation (e.g. SOX1, BRACH T, ELF5). In conclusion, the short term stabilization of HIF mediated by the PHD inhibitors JNJ and DMOG and hypoxia did not prevent the spontaneous loss of pluripotency markers in mESC. However, it significantly downregulated the proliferation of these cells. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  13. Chorionic villi derived mesenchymal like stem cells and expression of embryonic stem cells markers during long-term culturing. (United States)

    Katsiani, E; Garas, A; Skentou, C; Tsezou, A; Messini, C I; Dafopoulos, K; Daponte, A; Messinis, I E


    Mesenchymal stem cells (MSCs) can be obtained from a variety of human tissues. MSCs derived from placental chorionic villi of the first trimester are likely to resemble, biologically, embryonic stem cells (ESC), due to the earlier development stage of placenta. In the present study long-term cultures of MSC-like cells were assessed in order to evaluate MSCs multipotent characteristics and molecular features during the period of culture. CV-cells obtained from 10 samples of chorionic villus displayed typical fibroblastoid morphology, undergone 20 passages during a period of 120 days, maintaining a stable karyotype throughout long term expansion. The cells were positive, for CD90, CD73, CD105, CD29, CD44, HLA ABC antigens and negative for CD14, CD34, AC133, and HLA DR antigens as resulted from the flow cytometry analysis. CV-cells were differentiated in adipocytes, osteoblasts, chondrocytes and neuronal cells under specific culture conditions. The expression of the ESC-gene markers POU5F1 (Oct-4) and NANOG was observed at earliest stages (4-12 passages) and not at the late stages (14-20 passages) by RT-PCR analysis. ZFP42 and SOX2 expression were not detected. Moreover, CV-cells were found to express GATA4 but not NES (Nestin). Chorionic villi-derived cells possess multipotent properties, display high proliferation rate and self-renew capacity, share common surface antigens with adult MSCs and express certain embryonics stem cells gene markers. These characteristics highlight chorionic villi as an attractive source of MSCs for the needs of regenerative medicine.

  14. Early Gene Expression in Wounded Human Keratinocytes Revealed by DNA Microarray Analysis

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    Pascal Barbry


    Full Text Available Wound healing involves several steps: spreading of the cells, migration and proliferation. We have profiled gene expression during the early events of wound healing in normal human keratinocytes with a home-made DNA microarray containing about 1000 relevant human probes. An original wounding machine was used, that allows the wounding of up to 40% of the surface of a confluent monolayer of cultured cells grown on a Petri dish (compared with 5% with a classical ‘scratch’ method. The two aims of the present study were: (a to validate a limited number of genes by comparing the expression levels obtained with this technique with those found in the literature; (b to combine the use of the wounding machine with DNA microarray analysis for large-scale detection of the molecular events triggered during the early stages of the wound-healing process. The time-courses of RNA expression observed at 0.5, 1.5, 3, 6 and 15 h after wounding for genes such as c-Fos, c-Jun, Egr1, the plasminogen activator PLAU (uPA and the signal transducer and transcription activator STAT3, were consistent with previously published data. This suggests that our methodologies are able to perform quantitative measurement of gene expression. Transcripts encoding two zinc finger proteins, ZFP36 and ZNF161, and the tumour necrosis factor α-induced protein TNFAIP3, were also overexpressed after wounding. The role of the p38 mitogen-activated protein kinase (p38MAPK in wound healing was shown after the inhibition of p38 by SB203580, but our results also suggest the existence of surrogate activating pathways.

  15. Mouse obesity network reconstruction with a variational Bayes algorithm to employ aggressive false positive control

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    Logsdon Benjamin A


    Full Text Available Abstract Background We propose a novel variational Bayes network reconstruction algorithm to extract the most relevant disease factors from high-throughput genomic data-sets. Our algorithm is the only scalable method for regularized network recovery that employs Bayesian model averaging and that can internally estimate an appropriate level of sparsity to ensure few false positives enter the model without the need for cross-validation or a model selection criterion. We use our algorithm to characterize the effect of genetic markers and liver gene expression traits on mouse obesity related phenotypes, including weight, cholesterol, glucose, and free fatty acid levels, in an experiment previously used for discovery and validation of network connections: an F2 intercross between the C57BL/6 J and C3H/HeJ mouse strains, where apolipoprotein E is null on the background. Results We identified eleven genes, Gch1, Zfp69, Dlgap1, Gna14, Yy1, Gabarapl1, Folr2, Fdft1, Cnr2, Slc24a3, and Ccl19, and a quantitative trait locus directly connected to weight, glucose, cholesterol, or free fatty acid levels in our network. None of these genes were identified by other network analyses of this mouse intercross data-set, but all have been previously associated with obesity or related pathologies in independent studies. In addition, through both simulations and data analysis we demonstrate that our algorithm achieves superior performance in terms of power and type I error control than other network recovery algorithms that use the lasso and have bounds on type I error control. Conclusions Our final network contains 118 previously associated and novel genes affecting weight, cholesterol, glucose, and free fatty acid levels that are excellent obesity risk candidates.

  16. Meta-analysis of differentiating mouse embryonic stem cell gene expression kinetics reveals early change of a small gene set.

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    Clive H Glover


    Full Text Available Stem cell differentiation involves critical changes in gene expression. Identification of these should provide endpoints useful for optimizing stem cell propagation as well as potential clues about mechanisms governing stem cell maintenance. Here we describe the results of a new meta-analysis methodology applied to multiple gene expression datasets from three mouse embryonic stem cell (ESC lines obtained at specific time points during the course of their differentiation into various lineages. We developed methods to identify genes with expression changes that correlated with the altered frequency of functionally defined, undifferentiated ESC in culture. In each dataset, we computed a novel statistical confidence measure for every gene which captured the certainty that a particular gene exhibited an expression pattern of interest within that dataset. This permitted a joint analysis of the datasets, despite the different experimental designs. Using a ranking scheme that favored genes exhibiting patterns of interest, we focused on the top 88 genes whose expression was consistently changed when ESC were induced to differentiate. Seven of these (103728_at, 8430410A17Rik, Klf2, Nr0b1, Sox2, Tcl1, and Zfp42 showed a rapid decrease in expression concurrent with a decrease in frequency of undifferentiated cells and remained predictive when evaluated in additional maintenance and differentiating protocols. Through a novel meta-analysis, this study identifies a small set of genes whose expression is useful for identifying changes in stem cell frequencies in cultures of mouse ESC. The methods and findings have broader applicability to understanding the regulation of self-renewal of other stem cell types.

  17. Cosmological Particle Data Compression in Practice (United States)

    Zeyen, M.; Ahrens, J.; Hagen, H.; Heitmann, K.; Habib, S.


    In cosmological simulations trillions of particles are handled and several terabytes of unstructured particle data are generated in each time step. Transferring this data directly from memory to disk in an uncompressed way results in a massive load on I/O and storage systems. Hence, one goal of domain scientists is to compress the data before storing it to disk while minimizing the loss of information. To prevent reading back uncompressed data from disk, this can be done in an in-situ process. Since the simulation continuously generates data, the available time for the compression of one time step is limited. Therefore, the evaluation of compression techniques has shifted from only focusing on compression rates to include run-times and scalability.In recent years several compression techniques for cosmological data have become available. These techniques can be either lossy or lossless, depending on the technique. For both cases, this study aims to evaluate and compare the state of the art compression techniques for unstructured particle data. This study focuses on the techniques available in the Blosc framework with its multi-threading support, the XZ Utils toolkit with the LZMA algorithm that achieves high compression rates, and the widespread FPZIP and ZFP methods for lossy compressions.For the investigated compression techniques, quantitative performance indicators such as compression rates, run-time/throughput, and reconstruction errors are measured. Based on these factors, this study offers a comprehensive analysis of the individual techniques and discusses their applicability for in-situ compression. In addition, domain specific measures are evaluated on the reconstructed data sets, and the relative error rates and statistical properties are analyzed and compared. Based on this study future challenges and directions in the compression of unstructured cosmological particle data were identified.

  18. Identification of two evolutionarily conserved 5' cis-elements involved in regulating spatiotemporal expression of Nolz-1 during mouse embryogenesis.

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    Sunny Li-Yun Chang

    Full Text Available Proper development of vertebrate embryos depends not only on the crucial funtions of key evolutionarily conserved transcriptional regulators, but also on the precisely spatiotemporal expression of these transcriptional regulators. The mouse Nolz-1/Znf503/Zfp503 gene is a mammalian member of the conserved zinc-finger containing NET family. The expression pattern of Nolz-1 in mouse embryos is highly correlated with that of its homologues in different species. To study the spatiotemporal regulation of Nolz-1, we first identified two evolutionarily conserved cis-elements, UREA and UREB, in 5' upstream regions of mouse Nolz-1 locus. We then generated UREA-LacZ and UREB-LacZ transgenic reporter mice to characterize the putative enhancer activity of UREA and UREB. The results indicated that both UREA and UREB contained tissue-specific enhancer activity for directing LacZ expression in selective tissue organs during mouse embryogensis. UREA directed LacZ expression preferentially in selective regions of developing central nervous system, including the forebrain, hindbrain and spinal cord, whereas UREB directed LacZ expression mainly in other developing tissue organs such as the Nolz-1 expressing branchial arches and its derivatives, the apical ectodermal ridge of limb buds and the urogenital tissues. Both UREA and UREB directed strong LacZ expression in the lateral plate mesoderm where endogenous Nolz-1 was also expressed. Despite that the LacZ expression pattern did not full recapitulated the endogenous Nolz-1 expression and some mismatched expression patterns were observed, co-expression of LacZ and Nolz-1 did occur in many cells of selective tissue organs, such as in the ventrolateral cortex and ventral spinal cord of UREA-LacZ embryos, and the urogenital tubes of UREB-LacZ embryos. Taken together, our study suggests that UREA and UREB may function as evolutionarily conserved cis-regulatory elements that coordinate with other cis-elements to regulate

  19. A Genome-Wide mQTL Analysis in Human Adipose Tissue Identifies Genetic Variants Associated with DNA Methylation, Gene Expression and Metabolic Traits.

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    Petr Volkov

    Full Text Available Little is known about the extent to which interactions between genetics and epigenetics may affect the risk of complex metabolic diseases and/or their intermediary phenotypes. We performed a genome-wide DNA methylation quantitative trait locus (mQTL analysis in human adipose tissue of 119 men, where 592,794 single nucleotide polymorphisms (SNPs were related to DNA methylation of 477,891 CpG sites, covering 99% of RefSeq genes. SNPs in significant mQTLs were further related to gene expression in adipose tissue and obesity related traits. We found 101,911 SNP-CpG pairs (mQTLs in cis and 5,342 SNP-CpG pairs in trans showing significant associations between genotype and DNA methylation in adipose tissue after correction for multiple testing, where cis is defined as distance less than 500 kb between a SNP and CpG site. These mQTLs include reported obesity, lipid and type 2 diabetes loci, e.g. ADCY3/POMC, APOA5, CETP, FADS2, GCKR, SORT1 and LEPR. Significant mQTLs were overrepresented in intergenic regions meanwhile underrepresented in promoter regions and CpG islands. We further identified 635 SNPs in significant cis-mQTLs associated with expression of 86 genes in adipose tissue including CHRNA5, G6PC2, GPX7, RPL27A, THNSL2 and ZFP57. SNPs in significant mQTLs were also associated with body mass index (BMI, lipid traits and glucose and insulin levels in our study cohort and public available consortia data. Importantly, the Causal Inference Test (CIT demonstrates how genetic variants mediate their effects on metabolic traits (e.g. BMI, cholesterol, high-density lipoprotein (HDL, hemoglobin A1c (HbA1c and homeostatic model assessment of insulin resistance (HOMA-IR via altered DNA methylation in human adipose tissue. This study identifies genome-wide interactions between genetic and epigenetic variation in both cis and trans positions influencing gene expression in adipose tissue and in vivo (dysmetabolic traits associated with the development of

  20. REX-1 expression and p38 MAPK activation status can determine proliferation/differentiation fates in human mesenchymal stem cells.

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    Dilli Ram Bhandari

    Full Text Available BACKGROUND: REX1/ZFP42 is a well-known embryonic stem cell (ESC marker. However, the role of REX1, itself, is relatively unknown because the function of REX1 has only been reported in the differentiation of ESCs via STAT signaling pathways. Human mesenchymal stem cells (hMSCs isolated from young tissues and cancer cells express REX1. METHODOLOGY/PRINCIPAL FINDING: Human umbilical cord blood-derived MSCs (hUCB-MSCs and adipose tissue-derived MSCs (hAD-MSCs strongly express REX1 and have a lower activation status of p38 MAPK, but bone marrow-derived MSCs (hBM-MSCs have weak REX1 expression and higher activation of p38 MAPK. These results indicated that REX1 expression in hMSCs was positively correlated with proliferation rates but inversely correlated with the phosphorylation of p38 MAPK. In hUCB-MSCs, the roles of REX1 and p38 MAPK were investigated, and a knockdown study was performed using a lentiviral vector-based small hairpin RNA (shRNA. After REX1 knockdown, decreased cell proliferation was observed. In REX1 knocked-down hUCB-MSCs, the osteogenic differentiation ability deteriorated, but the adipogenic potential increased or was similar to that observed in the controls. The phosphorylation of p38 MAPK in hUCB-MSCs significantly increased after REX1 knockdown. After p38 MAPK inhibitor treatment, the cell growth in REX1 knocked-down hUCB-MSCs almost recovered, and the suppressed expression levels of CDK2 and CCND1 were also restored. The expression of MKK3, an upstream regulator of p38 MAPK, significantly increased in REX1 knocked-down hUCB-MSCs. The direct binding of REX1 to the MKK3 gene was confirmed by a chromatin immunoprecipitation (ChIP assay. CONCLUSIONS/SIGNIFICANCE: These findings showed that REX1 regulates the proliferation/differentiation of hMSCs through the suppression of p38 MAPK signaling via the direct suppression of MKK3. Therefore, p38 MAPK and REX-1 status can determine the cell fate of adult stem cells (ASCs. These

  1. Drought Response in Wheat: Key Genes and Regulatory Mechanisms Controlling Root System Architecture and Transpiration Efficiency

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    Manoj Kulkarni


    Full Text Available Abiotic stresses such as, drought, heat, salinity, and flooding threaten global food security. Crop genetic improvement with increased resilience to abiotic stresses is a critical component of crop breeding strategies. Wheat is an important cereal crop and a staple food source globally. Enhanced drought tolerance in wheat is critical for sustainable food production and global food security. Recent advances in drought tolerance research have uncovered many key genes and transcription regulators governing morpho-physiological traits. Genes controlling root architecture and stomatal development play an important role in soil moisture extraction and its retention, and therefore have been targets of molecular breeding strategies for improving drought tolerance. In this systematic review, we have summarized evidence of beneficial contributions of root and stomatal traits to plant adaptation to drought stress. Specifically, we discuss a few key genes such as, DRO1 in rice and ERECTA in Arabidopsis and rice that were identified to be the enhancers of drought tolerance via regulation of root traits and transpiration efficiency. Additionally, we highlight several transcription factor families, such as, ERF (ethylene response factors, DREB (dehydration responsive element binding, ZFP (zinc finger proteins, WRKY, and MYB that were identified to be both positive and negative regulators of drought responses in wheat, rice, maize, and/or Arabidopsis. The overall aim of this review is to provide an overview of candidate genes that have been identified as regulators of drought response in plants. The lack of a reference genome sequence for wheat and non-transgenic approaches for manipulation of gene functions in wheat in the past had impeded high-resolution interrogation of functional elements, including genes and QTLs, and their application in cultivar improvement. The recent developments in wheat genomics and reverse genetics, including the availability of a

  2. Microarray profiling of mononuclear peripheral blood cells identifies novel candidate genes related to chemoradiation response in rectal cancer.

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    Pablo Palma

    Full Text Available Preoperative chemoradiation significantly improves oncological outcome in locally advanced rectal cancer. However there is no effective method of predicting tumor response to chemoradiation in these patients. Peripheral blood mononuclear cells have emerged recently as pathology markers of cancer and other diseases, making possible their use as therapy predictors. Furthermore, the importance of the immune response in radiosensivity of solid organs led us to hypothesized that microarray gene expression profiling of peripheral blood mononuclear cells could identify patients with response to chemoradiation in rectal cancer. Thirty five 35 patients with locally advanced rectal cancer were recruited initially to perform the study. Peripheral blood samples were obtained before neaodjuvant treatment. RNA was extracted and purified to obtain cDNA and cRNA for hybridization of microarrays included in Human WG CodeLink bioarrays. Quantitative real time PCR was used to validate microarray experiment data. Results were correlated with pathological response, according to Mandard´s criteria and final UICC Stage (patients with tumor regression grade 1-2 and downstaging being defined as responders and patients with grade 3-5 and no downstaging as non-responders. Twenty seven out of 35 patients were finally included in the study. We performed a multiple t-test using Significance Analysis of Microarrays, to find those genes differing significantly in expression, between responders (n = 11 and non-responders (n = 16 to CRT. The differently expressed genes were: BC 035656.1, CIR, PRDM2, CAPG, FALZ, HLA-DPB2, NUPL2, and ZFP36. The measurement of FALZ (p = 0.029 gene expression level determined by qRT-PCR, showed statistically significant differences between the two groups. Gene expression profiling reveals novel genes in peripheral blood samples of mononuclear cells that could predict responders and non-responders to chemoradiation in patients with

  3. Impact of benzo(a)pyrene, Cu and their mixture on the proteomic response of Mytilus galloprovincialis

    International Nuclear Information System (INIS)

    Maria, V.L.; Gomes, T.; Barreira, L.; Bebianno, M.J.


    Highlights: •Distinct protein expression profiles dependent of BaP and Cu accumulation, metabolism and chemical interactions in mussels, Mytilus galloprovincialis. •Processes that involve adhesion and motility, cytoskeleton and cell structure, stress response, transcription regulation and energy metabolism are common mechanisms. •Traditional (ATP synthase, GST, HSP and actin) and novel biomarkers for BaP (ZFP), Cu (chitin synthase) and mixture (MVP) exposures identified in mussels. -- Abstract: In natural waters, chemical interactions between mixtures of contaminants can result in potential synergistic and/or antagonic effects in aquatic animals. Benzo(a)pyrene (BaP) and copper (Cu) are two widespread environmental contaminants with known toxicity towards mussels Mytilus spp. The effects of the individual and the interaction of BaP and Cu exposures were assessed in mussels Mytilus galloprovincialis using proteomic analysis. Mussels were exposed to BaP [10 μg L −1 (0.396 μM)], and Cu [10 μg L −1 (0.16 μM)], as well as to their binary mixture (mixture) for a period of 7 days. Proteomic analysis showed different protein expression profiles associated to each selected contaminant condition. A non-additive combined effect was observed in mixture in terms of new and suppressed proteins. Proteins more drastically altered (new, suppressed and 2-fold differentially expressed) were excised and analyzed by mass spectrometry, and eighteen putatively identified. Protein identification demonstrated the different accumulation, metabolism and chemical interactions of BaP, Cu and their mixture, resulting in different modes of action. Proteins associated with adhesion and motility (catchin, twitchin and twitchin-like protein), cytoskeleton and cell structure (α-tubulin and actin), stress response (heat shock cognate 71, heat shock protein 70, putative C1q domain containing protein), transcription regulation (zinc-finger BED domain-containing and nuclear receptor

  4. HITS-CLIP analysis uncovers a link between the Kaposi's sarcoma-associated herpesvirus ORF57 protein and host pre-mRNA metabolism.

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    Emi Sei


    Full Text Available The Kaposi's sarcoma associated herpesvirus (KSHV is an oncogenic virus that causes Kaposi's sarcoma, primary effusion lymphoma (PEL, and some forms of multicentric Castleman's disease. The KSHV ORF57 protein is a conserved posttranscriptional regulator of gene expression that is essential for virus replication. ORF57 is multifunctional, but most of its activities are directly linked to its ability to bind RNA. We globally identified virus and host RNAs bound by ORF57 during lytic reactivation in PEL cells using high-throughput sequencing of RNA isolated by cross-linking immunoprecipitation (HITS-CLIP. As expected, ORF57-bound RNA fragments mapped throughout the KSHV genome, including the known ORF57 ligand PAN RNA. In agreement with previously published ChIP results, we observed that ORF57 bound RNAs near the oriLyt regions of the genome. Examination of the host RNA fragments revealed that a subset of the ORF57-bound RNAs was derived from transcript 5' ends. The position of these 5'-bound fragments correlated closely with the 5'-most exon-intron junction of the pre-mRNA. We selected four candidates (BTG1, EGR1, ZFP36, and TNFSF9 and analyzed their pre-mRNA and mRNA levels during lytic phase. Analysis of both steady-state and newly made RNAs revealed that these candidate ORF57-bound pre-mRNAs persisted for longer periods of time throughout infection than control RNAs, consistent with a role for ORF57 in pre-mRNA metabolism. In addition, exogenous expression of ORF57 was sufficient to increase the pre-mRNA levels and, in one case, the mRNA levels of the putative ORF57 targets. These results demonstrate that ORF57 interacts with specific host pre-mRNAs during lytic reactivation and alters their processing, likely by stabilizing pre-mRNAs. These data suggest that ORF57 is involved in modulating host gene expression in addition to KSHV gene expression during lytic reactivation.

  5. Impact of benzo(a)pyrene, Cu and their mixture on the proteomic response of Mytilus galloprovincialis

    Energy Technology Data Exchange (ETDEWEB)

    Maria, V.L., E-mail: [CIMA, Faculty of Sciences and Technology, University of Algarve, Campus de Gambelas, 8005-139 Faro (Portugal); Gomes, T., E-mail: [CIMA, Faculty of Sciences and Technology, University of Algarve, Campus de Gambelas, 8005-139 Faro (Portugal); Barreira, L., E-mail: [CCMAR, Faculty of Sciences and Technology, University of Algarve, Campus de Gambelas, 8005-139 Faro (Portugal); Bebianno, M.J., E-mail: [CIMA, Faculty of Sciences and Technology, University of Algarve, Campus de Gambelas, 8005-139 Faro (Portugal)


    Highlights: •Distinct protein expression profiles dependent of BaP and Cu accumulation, metabolism and chemical interactions in mussels, Mytilus galloprovincialis. •Processes that involve adhesion and motility, cytoskeleton and cell structure, stress response, transcription regulation and energy metabolism are common mechanisms. •Traditional (ATP synthase, GST, HSP and actin) and novel biomarkers for BaP (ZFP), Cu (chitin synthase) and mixture (MVP) exposures identified in mussels. -- Abstract: In natural waters, chemical interactions between mixtures of contaminants can result in potential synergistic and/or antagonic effects in aquatic animals. Benzo(a)pyrene (BaP) and copper (Cu) are two widespread environmental contaminants with known toxicity towards mussels Mytilus spp. The effects of the individual and the interaction of BaP and Cu exposures were assessed in mussels Mytilus galloprovincialis using proteomic analysis. Mussels were exposed to BaP [10 μg L{sup −1} (0.396 μM)], and Cu [10 μg L{sup −1} (0.16 μM)], as well as to their binary mixture (mixture) for a period of 7 days. Proteomic analysis showed different protein expression profiles associated to each selected contaminant condition. A non-additive combined effect was observed in mixture in terms of new and suppressed proteins. Proteins more drastically altered (new, suppressed and 2-fold differentially expressed) were excised and analyzed by mass spectrometry, and eighteen putatively identified. Protein identification demonstrated the different accumulation, metabolism and chemical interactions of BaP, Cu and their mixture, resulting in different modes of action. Proteins associated with adhesion and motility (catchin, twitchin and twitchin-like protein), cytoskeleton and cell structure (α-tubulin and actin), stress response (heat shock cognate 71, heat shock protein 70, putative C1q domain containing protein), transcription regulation (zinc-finger BED domain-containing and nuclear

  6. Repositioning of Memantine as a Potential Novel Therapeutic Agent against Meningitic E. coli-Induced Pathogenicities through Disease-Associated Alpha7 Cholinergic Pathway and RNA Sequencing-Based Transcriptome Analysis of Host Inflammatory Responses.

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    Jing-Yi Yu

    Full Text Available Neonatal sepsis and meningitis (NSM remains a leading cause worldwide of mortality and morbidity in newborn infants despite the availability of antibiotics over the last several decades. E. coli is the most common gram-negative pathogen causing NSM. Our previous studies show that α7 nicotinic receptor (α7 nAChR, an essential regulator of inflammation, plays a detrimental role in the host defense against NSM. Despite notable successes, there still exists an unmet need for new effective therapeutic approaches to treat this disease. Using the in vitro/in vivo models of the blood-brain barrier (BBB and RNA-seq, we undertook a drug repositioning study to identify unknown antimicrobial activities for known drugs. We have demonstrated for the first time that memantine (MEM, a FDA-approved drug for treatment of Alzheimer's disease, could very efficiently block E. coli-caused bacteremia and meningitis in a mouse model of NSM in a manner dependent on α7 nAChR. MEM was able to synergistically enhance the antibacterial activity of ampicillin in HBMEC infected with E. coli K1 (E44 and in neonatal mice with E44-caused bacteremia and meningitis. Differential gene expression analysis of RNA-Seq data from mouse BMEC infected with E. coli K1 showed that several E44-increased inflammatory factors, including IL33, IL18rap, MMP10 and Irs1, were significantly reduced by MEM compared to the infected cells without drug treatment. MEM could also significantly up-regulate anti-inflammatory factors, including Tnfaip3, CISH, Ptgds and Zfp36. Most interestingly, these factors may positively and negatively contribute to regulation of NF-κB, which is a hallmark feature of bacterial meningitis. Furthermore, we have demonstrated that circulating BMEC (cBMEC are the potential novel biomarkers for NSM. MEM could significantly reduce E44-increased blood level of cBMEC in mice. Taken together, our data suggest that memantine can efficiently block host inflammatory responses to

  7. Contribution of Heritability and Epigenetic Factors to Skeletal Muscle Mass Variation in United Kingdom Twins. (United States)

    Livshits, Gregory; Gao, Fei; Malkin, Ida; Needhamsen, Maria; Xia, Yudong; Yuan, Wei; Bell, Christopher G; Ward, Kirsten; Liu, Yuan; Wang, Jun; Bell, Jordana T; Spector, Tim D


    Skeletal muscle mass (SMM) is one of the major components of human body composition, with deviations from normal values often leading to sarcopenia. Our major aim was to conduct a genome-wide DNA methylation study in an attempt to identify potential genomic regions associated with SMM. This was a mixed cross-sectional and longitudinal study. Community-based study. A total of 1550 middle-aged United Kingdom twins (monozygotic [MZ] and dizygotic [DZ]), 297 of which were repeatedly measured participated in the study. Appendicular lean mass assessed using dual-energy X-ray absorptiometry technology, and methylated DNA immunoprecipitation sequencing DNA methylation profiling genome-wide were obtained from each individual. Heritability estimate of SMM, with simultaneous adjustment for covariates obtained using variance decomposition analysis, was h(2) = 0.809 ± 0.050. After quality control and analysis of longitudinal stability, the DNA methylation data comprised of 723 029 genomic sites, with positive correlations between repeated measurements (Rrepeated = 0.114-0.905). Correlations between MZ and DZ twins were 0.51 and 0.38 at a genome-wide average, respectively, and clearly increased with Rrepeated. Testing for DNA methylation association with SMM in 50 discordant MZ twins revealed 36 081 nominally significant results, of which the top-ranked 134 signals (P 0.40) were subjected to replication in the sample of 1196 individuals. Seven SMM methylation association signals replicated at a false discovery rate less than 0.1, and these were located in or near genes DNAH12, CAND1, CYP4F29P, and ZFP64, which have previously been highlighted in muscle-related studies. Adjusting for age, smoking, and blood cell heterogeneity did not alter significance of these associations. This epigenome-wide study, testing longitudinally stable methylation sites, discovered and replicated a number of associations between DNA methylation at CpG loci and SMM. Four replicated signals were