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Sample records for apex gene knockout

  1. The mammalian gene function resource: The International Knockout Mouse Consortium

    NARCIS (Netherlands)

    A. Bradley (Allan); K. Anastassiadis (Konstantinos); A. Ayadi (Abdelkader); J.F. Battey (James); C. Bell (Cindy); M.-C. Birling (Marie-Christine); J. Bottomley (Joanna); S.D.M. Brown (Steve); F. Bürger (Friederike); C.J. Bult (Carol); W. Bushell (Wendy); F.S. Collins (Francis); C. Desaintes (Christian); B. Doe (Brendan); E. Aris (Economides); J.T. Eppig (Janan); R.H. Finnell (Richard); C. Fletcher (Colin); M. Fray (Martin); D. Frendewey (David); R.H. Friedel (Roland); F.G. Grosveld (Frank); J. Hansen; Y. Hérault (Yann); G. Hicks (Geoffrey); A. Hörlein (Andreas); C. Houghton (Catherine); M. Hrabé De Angelis (Martin); D. Huylebroeck (Danny); V. Iyer (Vivek); P.J. de Jong (Pieter); J.A. Kadin (James); C. Kaloff (Cornelia); K. Kennedy (Karen); M. Koutsourakis (Manousos); K.C. Kent Lloyd (K.); S. Marschall (Susan); J. Mason (Jeremy); C. McKerlie (Colin); M.P. McLeod (Michael); H. von Melchner (Harald); M. Moore (Matt); A.O. Mujica (Alejandro); A. Nagy (Andras); M. Nefedov (Mikhail); L.M. Nutter (Lauryl); G. Pavlovic (Guillaume); J.L. Peterson (Jane); I. Pollock; R. Ramirez-Solis (Ramiro); D.E. Rancourt (Derrick); M. Raspa (Marcello); J.E. Remacle (Jacques); M. Ringwald (Martin); B. Rosen (Barry); N. Rosenthal (Nadia); J. Rossant (Janet); P. Ruiz Noppinger (Patricia); S. Ryder; J.Z. Schick (Joel Zupicich); F. Schnütgen (Frank); C.J. Schofield (Christopher); C. Seisenberger (Claudia); M. Selloum (Mohammed); E.M. Simpson (Elizabeth); W.C. Skarnes (William); D. Smedley (Damian); W.L. Stanford (William); A. Francis Stewart (A.); K. Stone (Kevin); K. Swan (Kate); H. Tadepally (Hamsa); J.L. Teboul (Jean Louis); G.P. Tocchini-Valentini (Glauco); D. Valenzuela (David); A.P. West (Anthony); K.-I. Yamamura (Ken-Ichi); Y. Yoshinaga (Yuko); M. Wurst (Martin)

    2012-01-01

    textabstractIn 2007, the International Knockout Mouse Consortium (IKMC) made the ambitious promise to generate mutations in virtually every protein-coding gene of the mouse genome in a concerted worldwide action. Now, 5 years later, the IKMC members have developed highthroughput gene trapping and, i

  2. Genomic structure of the rat major AP endonuclease gene (Apex with an adjacent putative O-sialoglycoprotease gene (Prsmg1/Gcpl1 and a processed Apex pseudogene (Apexp1.

    Directory of Open Access Journals (Sweden)

    Yao,Ming

    1999-12-01

    Full Text Available Genomic sequencing and chromosomal assignment of the gene encoding rat APEX nuclease, a multifunctional DNA repair enzyme, were performed. An active Apex gene and a processed pseudogene were isolated from a rat genomic library. The active Apex gene consists of 5 exons and 4 introns spanning 2.1 kb. The putative promoter region of the Apex gene lacks the typical TATA box, but contains CAAT boxes and a CpG island having putative binding sites for several transcription factors, such as Sp1, AP-2, GATA-1 and ATF. A putative O-sialoglycoprotease (a homologue of Pasteurella haemolytica glycoprotease, gcp; abbreviated as Prsmg1/Gcpl1 gene consisting of 11 exons and 10 introns spanning 7.3 kb lies immediately adjacent to the Apex gene in a 5'-to-5' orientation. The Apex gene locus was mapped to rat chromosome 15p12 using in situ hybridization. The processed pseudogene (designated as rat Apexp1 has a nucleotide sequence 87.1% identical to that of the rat Apex cDNA, although several stop codons interrupting the coding sequences and multiple nucleotide deletions were observed. The Apexp1 is located in an inactive LINE sequence. Calculation of nucleotide substitution rates suggests that the immediate, active progenitor of Apexp1 arose 23 million years ago and that the non-functionalization occurred 15 million years ago.

  3. Efficient Gene Knockout in Goats Using CRISPR/Cas9 System

    OpenAIRE

    Ni, Wei; Qiao, Jun; Hu, Shengwei; Zhao, Xinxia; Regouski, Misha; Yang, Min; Polejaeva, Irina A.; Chen, Chuangfu

    2014-01-01

    The CRISPR/Cas9 system has been adapted as an efficient genome editing tool in laboratory animals such as mice, rats, zebrafish and pigs. Here, we report that CRISPR/Cas9 mediated approach can efficiently induce monoallelic and biallelic gene knockout in goat primary fibroblasts. Four genes were disrupted simultaneously in goat fibroblasts by CRISPR/Cas9-mediated genome editing. The single-gene knockout fibroblasts were successfully used for somatic cell nuclear transfer (SCNT) and resulted i...

  4. Production of p53 gene knockout rats by homologous recombination in embryonic stem cells

    OpenAIRE

    Tong, Chang; Li, Ping; Wu, Nancy L; Yan, Youzhen; Ying, Qi-Long

    2010-01-01

    The use of homologous recombination to modify genes in embryonic stem (ES) cells provides a powerful means to elucidate gene function and create disease models1. Application of this technology to engineer genes in rats has previously been impossible in the absence of germline competent ES cells in this species. We have recently established authentic rat ES cells2, 3. Here we report the generation of the first gene knockout rats using the ES cell-based gene targeting technology. We designed a ...

  5. Effects of alpha-AMPK knockout on exercise-induced gene activation in mouse skeletal muscle

    DEFF Research Database (Denmark)

    Jørgensen, Sebastian Beck; Wojtaszewski, Jørgen; Viollet, Benoit;

    2005-01-01

    We tested the hypothesis that 5'AMP-activated protein kinase (AMPK) plays an important role in regulating the acute, exercise-induced activation of metabolic genes in skeletal muscle, which were dissected from whole-body a2- and a1-AMPK knockout (KO) and wild-type (WT) mice at rest, after treadmi...

  6. Hitting two birds with one stone: The unforeseen consequences of nested gene knockouts in Caenorhabditis elegans.

    Science.gov (United States)

    Jovelin, Richard; Cutter, Asher D

    2016-01-01

    Nested genes represent an intriguing form of non-random genomic organization in which the boundaries of one gene are fully contained within another, longer host gene. The C. elegans genome contains over 10,000 nested genes, 92% of which are ncRNAs, which occur inside 16% of the protein coding gene complement. Host genes are longer than non-host coding genes, owing to their longer and more numerous introns. Indel alleles are available for nearly all of these host genes that simultaneously alter the nested gene, raising the possibility of nested gene disruption contributing to phenotypes that might be attributed to the host gene. Such dual-knockouts could represent a source of misinterpretation about host gene function. Dual-knockouts might also provide a novel source of synthetic phenotypes that reveal the functional effects of ncRNA genes, whereby the host gene disruption acts as a perturbed genetic background to help unmask ncRNA phenotypes. PMID:27386165

  7. Have gene knockouts caused evolutionary reversals in the mammalian first arch?

    Science.gov (United States)

    Smith, K K; Schneider, R A

    1998-03-01

    Many recent gene knockout experiments cause anatomical changes to the jaw region of mice that several investigators claim are evolutionary reversals. Here we evaluate these mutant phenotypes and the assertions of atavism. We argue that following the knockout of Hoxa-2, Dlx-2, MHox, Otx2, and RAR genes, ectopic cartilages arise as secondary consequences of disruptions in normal processes of cell specification, migration, or differentiation. These disruptions cause an excess of mesenchyme to accumulate in a region through which skeletal progenitor cells usually migrate, and at a site of condensation that is normally present in mammals but that is too small to chondrify. We find little evidence that these genes, when disrupted, cause a reversion to any primitive condition and although changes in their expression may have played a role in the evolution of the mammalian jaw, their function during morphogenesis is not sufficiently understood to confirm such hypotheses. PMID:9631652

  8. Germ line knockout of IGFBP-3 reveals influences of the gene on mammary gland neoplasia.

    Science.gov (United States)

    Blouin, Marie-José; Bazile, Miguel; Birman, Elena; Zakikhani, Mahvash; Florianova, Livia; Aleynikova, Olga; Powell, David R; Pollak, Michael

    2015-02-01

    Insulin-like growth factor binding protein-3 (IGFBP-3) is an important carrier protein for insulin-like growth factors (IGFs) in the circulation. IGFBP-3 antagonizes the growth-promoting and anti-apoptotic activities of IGFs in experimental systems, but in certain contexts can increase IGF bioactivity, probably by increasing its half-life. The goal of this study was to investigate the role of IGFBP-3 in breast carcinogenesis and breast cancer metastasis. In the first part of the study, we exposed IGFBP-3 knockout and wild-type female mice to dimethylbenz[a]anthracene (DMBA) and followed them for appearance of primary tumors for up to 13 months. In the second part, mice of each genotype received an IV injection of 4T1 mammary carcinoma cells and then lung nodules were counted. Our results show that IGFBP-3 knockout mice developed breast tumors significantly earlier than the wild-type (13.9 ± 1.1 versus 22.5 ± 3.3 weeks, respectively, P = 0.0144), suggesting tumor suppression activity of IGFBP-3. In tumors of IGFBP-3 knockout mice, levels of phospho-AKT(Ser473) were increased compared to wild-type mice. The lung metastasis assay showed significantly more and larger lung nodules in IGFBP-3 knockout mice than in wild-type mice. While we observed increased levels of IGFBP-5 protein in the IGFBP-3 knockout mice, our findings suggest that this was not sufficient to completely compensate for the absence of IGFBP-3. Even though knockout of IGFBP-3 is associated with only a subtle phenotype under control conditions, our results reveal that loss of this gene has measurable effects on breast carcinogenesis and breast cancer metastasis. PMID:25614235

  9. Unintentional miRNA ablation is a risk factor in gene knockout studies: a short report.

    Directory of Open Access Journals (Sweden)

    Ivan Osokine

    2008-02-01

    Full Text Available One of the most powerful techniques for studying the function of a gene is to disrupt the expression of that gene using genetic engineering strategies such as targeted recombination or viral integration of gene trap cassettes. The tremendous utility of these tools was recognized this year with the awarding of the Nobel Prize in Physiology or Medicine to Capecchi, Evans, and Smithies for their pioneering work in targeted recombination mutagenesis in mammals. Another noteworthy discovery made nearly a decade ago was the identification of a novel class of non-coding genes called microRNAs. MicroRNAs are among the largest known classes of regulatory elements with more than 1000 predicted to exist in the mouse genome. Over 50% of known microRNAs are located within introns of coding genes. Given that currently about half of the genes in mouse have been knocked out, we investigated the possibility that intronic microRNAs may have been coincidentally deleted or disrupted in some of these mouse models. We searched published murine knockout studies and gene trap embryonic stem cell line databases for cases where a microRNA was located within or near the manipulated genomic loci, finding almost 200 cases where microRNA expression may have been disrupted along with another gene. Our results draw attention to the need for careful planning in future knockout studies to minimize the unintentional disruption of microRNAs. These data also raise the possibility that many knockout studies may need to be reexamined to determine if loss of a microRNA contributes to the phenotypic consequences attributed to loss of a protein-encoding gene.

  10. Outcome of urogenital infection with Chlamydia muridarum in CD-14 gene knockout mice

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    Ramsey Kyle H

    2006-09-01

    Full Text Available Abstract Background CD14 has been postulated to play a role in chlamydial immunity and immunopathology. There is evidence to support this role in human infections but its function in a mouse model has not been investigated. Methods Female CD14 gene knockout and C57BL/6J wild type mice were infected intravaginally with Chlamydia muridarum. The infection course was monitored by detection of viable chlamydiae from serially collected cervical-vaginal swabs. The sequela of tubal factor infertility was assessed using hydrosalpinx formation as a surrogate marker. Results A significantly abbreviated infection course was observed in the CD14 gene knockout mice but hydrosalpinx formation occurred at similar rates between the two groups. Conclusion Involvement of CD14 during chlamydial infection impedes infection resolution but this does not affect the sequela of infertility as assessed by hydrosalpinx formation.

  11. Generating Targeted Gene Knockout Lines in Physcomitrella patens to Study Evolution of Stress-Responsive Mechanisms

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    Maronova, Monika; Kalyna, Maria

    2016-01-01

    The moss Physcomitrella patens possesses highly efficient homologous recombination allowing targeted gene manipulations and displays many features of the early land plants including high tolerance to abiotic stresses. It is therefore an invaluable model organism for studies of gene functions and comparative studies of evolution of stress responses in plants. Here, we describe a method for generating targeted gene knockout lines in P. patens using a polyethylene glycol-mediated transformation of protoplasts including basic in vitro growth, propagation, and maintenance techniques. PMID:26867627

  12. Agrobacterium-mediated transformation of Eucalyptus globulus using explants with shoot apex with introduction of bacterial choline oxidase gene to enhance salt tolerance.

    Science.gov (United States)

    Matsunaga, Etsuko; Nanto, Kazuya; Oishi, Masatoshi; Ebinuma, Hiroyasu; Morishita, Yoshihiko; Sakurai, Nozomu; Suzuki, Hideyuki; Shibata, Daisuke; Shimada, Teruhisa

    2012-01-01

    Eucalyptus globulus is one of the most economically important plantation hardwoods for paper making. However, its low transformation frequency has prevented genetic engineering of this species with useful genes. We found the hypocotyl section with a shoot apex has the highest regeneration ability among another hypocotyl sections, and have developed an efficient Agrobacterium-mediated transformation method using these materials. We then introduced a salt tolerance gene, namely a bacterial choline oxidase gene (codA) with a GUS reporter gene, into E. globulus. The highest frequency of transgenic shoot regeneration from hypocotyls with shoot apex was 7.4% and the average frequency in four experiments was 4.0%, 12-fold higher than that from hypocotyls without shoot apex. Using about 10,000 explants, over 250 regenerated buds were confirmed as transformants by GUS analysis. Southern blot analysis of 100 elongated shoots confirmed successful generation of stable transformants. Accumulation of glycinebetaine was investigated in 44 selected transgenic lines, which showed 1- to 12-fold higher glycinebetaine levels than non-transgenic controls. Rooting of 16 transgenic lines was successful using a photoautotrophic method under enrichment with 1,000 ppm CO(2). The transgenic whole plantlets were transplanted into potting soil and grown normally in a growth room. They showed salt tolerance to 300 mM NaCl. The points of our system are using explants with shoot apex as materials, inhibiting the elongation of the apex on the selection medium, and regenerating transgenic buds from the side opposite to the apex. This approach may also solve transformation problems in other important plants.

  13. Phenotypes Associated with Knockouts of Eight Dense Granule Gene Loci (GRA2-9) in Virulent Toxoplasma gondii.

    Science.gov (United States)

    Rommereim, Leah M; Bellini, Valeria; Fox, Barbara A; Pètre, Graciane; Rak, Camille; Touquet, Bastien; Aldebert, Delphine; Dubremetz, Jean-François; Cesbron-Delauw, Marie-France; Mercier, Corinne; Bzik, David J

    2016-01-01

    Toxoplasma gondii actively invades host cells and establishes a parasitophorous vacuole (PV) that accumulates many proteins secreted by the dense granules (GRA proteins). To date, at least 23 GRA proteins have been reported, though the function(s) of most of these proteins still remains unknown. We targeted gene knockouts at ten GRA gene loci (GRA1-10) to investigate the cellular roles and essentiality of these classical GRA proteins during acute infection in the virulent type I RH strain. While eight of these genes (GRA2-9) were successfully knocked out, targeted knockouts at the GRA1 and GRA10 loci were not obtained, suggesting these GRA proteins may be essential. As expected, the Δgra2 and Δgra6 knockouts failed to form an intravacuolar network (IVN). Surprisingly, Δgra7 exhibited hyper-formation of the IVN in both normal and lipid-free growth conditions. No morphological alterations were identified in parasite or PV structures in the Δgra3, Δgra4, Δgra5, Δgra8, or Δgra9 knockouts. With the exception of the Δgra3 and Δgra8 knockouts, all of the GRA knockouts exhibited defects in their infection rate in vitro. While the single GRA knockouts did not exhibit reduced replication rates in vitro, replication rate defects were observed in three double GRA knockout strains (Δgra4Δgra6, Δgra3Δgra5 and Δgra3Δgra7). However, the virulence of single or double GRA knockout strains in CD1 mice was not affected. Collectively, our results suggest that while the eight individual GRA proteins investigated in this study (GRA2-9) are not essential, several GRA proteins may provide redundant and potentially important functions during acute infection. PMID:27458822

  14. Knockout of exogenous EGFP gene in porcine somatic cells using zinc-finger nucleases

    International Nuclear Information System (INIS)

    Research highlights: → EGFP gene integrated in porcine somatic cells could be knocked out using the ZFN-KO system. → ZFNs induced targeted mutations in porcine primary cultured cells. → Complete absence of EGFP fluorescence was confirmed in ZFN-treated cells. -- Abstract: Zinc-finger nucleases (ZFNs) are expected as a powerful tool for generating gene knockouts in laboratory and domestic animals. Currently, it is unclear whether this technology can be utilized for knocking-out genes in pigs. Here, we investigated whether knockout (KO) events in which ZFNs recognize and cleave a target sequence occur in porcine primary cultured somatic cells that harbor the exogenous enhanced green fluorescent protein (EGFP) gene. ZFN-encoding mRNA designed to target the EGFP gene was introduced by electroporation into the cell. Using the Surveyor nuclease assay and flow cytometric analysis, we confirmed ZFN-induced cleavage of the target sequence and the disappearance of EGFP fluorescence expression in ZFN-treated cells. In addition, sequence analysis revealed that ZFN-induced mutations such as base substitution, deletion, or insertion were generated in the ZFN cleavage site of EGFP-expression negative cells that were cloned from ZFN-treated cells, thereby showing it was possible to disrupt (i.e., knock out) the function of the EGFP gene in porcine somatic cells. To our knowledge, this study provides the first evidence that the ZFN-KO system can be applied to pigs. These findings may open a new avenue to the creation of gene KO pigs using ZFN-treated cells and somatic cell nuclear transfer.

  15. Knockout of exogenous EGFP gene in porcine somatic cells using zinc-finger nucleases

    Energy Technology Data Exchange (ETDEWEB)

    Watanabe, Masahito [Japan Science and Technology Agency (JST), ERATO, Nakauchi Stem Cell and Organ Regeneration Project, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639 (Japan); Department of Life Sciences, School of Agriculture, Meiji University, 1-1-1 Higashimita, Tama-ku, Kawasaki, Kanagawa 214-8571 (Japan); Umeyama, Kazuhiro [Department of Life Sciences, School of Agriculture, Meiji University, 1-1-1 Higashimita, Tama-ku, Kawasaki, Kanagawa 214-8571 (Japan); International Cluster for Bio-Resource Research, Meiji University, 1-1-1 Higashimita, Tama-ku, Kawasaki, Kanagawa 214-8571 (Japan); Matsunari, Hitomi [Department of Life Sciences, School of Agriculture, Meiji University, 1-1-1 Higashimita, Tama-ku, Kawasaki, Kanagawa 214-8571 (Japan); Takayanagi, Shuko [Japan Science and Technology Agency (JST), ERATO, Nakauchi Stem Cell and Organ Regeneration Project, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639 (Japan); Department of Life Sciences, School of Agriculture, Meiji University, 1-1-1 Higashimita, Tama-ku, Kawasaki, Kanagawa 214-8571 (Japan); Haruyama, Erika; Nakano, Kazuaki; Fujiwara, Tsukasa; Ikezawa, Yuka [Department of Life Sciences, School of Agriculture, Meiji University, 1-1-1 Higashimita, Tama-ku, Kawasaki, Kanagawa 214-8571 (Japan); Nakauchi, Hiromitsu [Japan Science and Technology Agency (JST), ERATO, Nakauchi Stem Cell and Organ Regeneration Project, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639 (Japan); Center for Stem Cell Biology and Regenerative Medicine, Institute of Medical Science, Tokyo University, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639 (Japan); and others

    2010-11-05

    Research highlights: {yields} EGFP gene integrated in porcine somatic cells could be knocked out using the ZFN-KO system. {yields} ZFNs induced targeted mutations in porcine primary cultured cells. {yields} Complete absence of EGFP fluorescence was confirmed in ZFN-treated cells. -- Abstract: Zinc-finger nucleases (ZFNs) are expected as a powerful tool for generating gene knockouts in laboratory and domestic animals. Currently, it is unclear whether this technology can be utilized for knocking-out genes in pigs. Here, we investigated whether knockout (KO) events in which ZFNs recognize and cleave a target sequence occur in porcine primary cultured somatic cells that harbor the exogenous enhanced green fluorescent protein (EGFP) gene. ZFN-encoding mRNA designed to target the EGFP gene was introduced by electroporation into the cell. Using the Surveyor nuclease assay and flow cytometric analysis, we confirmed ZFN-induced cleavage of the target sequence and the disappearance of EGFP fluorescence expression in ZFN-treated cells. In addition, sequence analysis revealed that ZFN-induced mutations such as base substitution, deletion, or insertion were generated in the ZFN cleavage site of EGFP-expression negative cells that were cloned from ZFN-treated cells, thereby showing it was possible to disrupt (i.e., knock out) the function of the EGFP gene in porcine somatic cells. To our knowledge, this study provides the first evidence that the ZFN-KO system can be applied to pigs. These findings may open a new avenue to the creation of gene KO pigs using ZFN-treated cells and somatic cell nuclear transfer.

  16. Improving cold storage and processing traits in potato through targeted gene knockout.

    Science.gov (United States)

    Clasen, Benjamin M; Stoddard, Thomas J; Luo, Song; Demorest, Zachary L; Li, Jin; Cedrone, Frederic; Tibebu, Redeat; Davison, Shawn; Ray, Erin E; Daulhac, Aurelie; Coffman, Andrew; Yabandith, Ann; Retterath, Adam; Haun, William; Baltes, Nicholas J; Mathis, Luc; Voytas, Daniel F; Zhang, Feng

    2016-01-01

    Cold storage of potato tubers is commonly used to reduce sprouting and extend postharvest shelf life. However, cold temperature stimulates the accumulation of reducing sugars in potato tubers. Upon high-temperature processing, these reducing sugars react with free amino acids, resulting in brown, bitter-tasting products and elevated levels of acrylamide--a potential carcinogen. To minimize the accumulation of reducing sugars, RNA interference (RNAi) technology was used to silence the vacuolar invertase gene (VInv), which encodes a protein that breaks down sucrose to glucose and fructose. Because RNAi often results in incomplete gene silencing and requires the plant to be transgenic, here we used transcription activator-like effector nucleases (TALENs) to knockout VInv within the commercial potato variety, Ranger Russet. We isolated 18 plants containing mutations in at least one VInv allele, and five of these plants had mutations in all VInv alleles. Tubers from full VInv-knockout plants had undetectable levels of reducing sugars, and processed chips contained reduced levels of acrylamide and were lightly coloured. Furthermore, seven of the 18 modified plant lines appeared to contain no TALEN DNA insertions in the potato genome. These results provide a framework for using TALENs to quickly improve traits in commercially relevant autotetraploid potato lines. PMID:25846201

  17. Global analysis of gene expression in the developing brain of Gtf2ird1 knockout mice.

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    Jennifer O'Leary

    Full Text Available BACKGROUND: Williams-Beuren Syndrome (WBS is a neurodevelopmental disorder caused by a hemizygous deletion of a 1.5 Mb region on chromosome 7q11.23 encompassing 26 genes. One of these genes, GTF2IRD1, codes for a putative transcription factor that is expressed throughout the brain during development. Genotype-phenotype studies in patients with atypical deletions of 7q11.23 implicate this gene in the neurological features of WBS, and Gtf2ird1 knockout mice show reduced innate fear and increased sociability, consistent with features of WBS. Multiple studies have identified in vitro target genes of GTF2IRD1, but we sought to identify in vivo targets in the mouse brain. METHODOLOGY/PRINCIPAL FINDINGS: We performed the first in vivo microarray screen for transcriptional targets of Gtf2ird1 in brain tissue from Gtf2ird1 knockout and wildtype mice at embryonic day 15.5 and at birth. Changes in gene expression in the mutant mice were moderate (0.5 to 2.5 fold and of candidate genes with altered expression verified using real-time PCR, most were located on chromosome 5, within 10 Mb of Gtf2ird1. siRNA knock-down of Gtf2ird1 in two mouse neuronal cell lines failed to identify changes in expression of any of the genes identified from the microarray and subsequent analysis showed that differences in expression of genes on chromosome 5 were the result of retention of that chromosome region from the targeted embryonic stem cell line, and so were dependent upon strain rather than Gtf2ird1 genotype. In addition, specific analysis of genes previously identified as direct in vitro targets of GTF2IRD1 failed to show altered expression. CONCLUSIONS/SIGNIFICANCE: We have been unable to identify any in vivo neuronal targets of GTF2IRD1 through genome-wide expression analysis, despite widespread and robust expression of this protein in the developing rodent brain.

  18. Revealing the essentiality of multiple archaeal pcna genes using a mutant propagation assay based on an improved knockout method

    DEFF Research Database (Denmark)

    Zhang, Changyi; Guo, Li; Deng, Ling;

    2010-01-01

    , a hyperthermophilic crenarchaeon but failed with two conventional knockout methods. Then, a new knockout scheme denoted the marker insertion and target gene deletion (MID) was developed with which transformants were obtained for each pMID-pcna plasmid. We found that mutant cells persisted in transformant cultures...... during incubation of pMID-pcna3 and pMID-araS-pcna1 transformants under counter selection. Studying the propagation of mutant cells by semi-quantitative PCR analysis of the deleted target gene allele (Deltapcna1 or Deltapcna3) revealed that mutant cells lost propagativity, demonstrating that these pcna...

  19. DNA-free two-gene knockout in Chlamydomonas reinhardtii via CRISPR-Cas9 ribonucleoproteins.

    Science.gov (United States)

    Baek, Kwangryul; Kim, Duk Hyoung; Jeong, Jooyeon; Sim, Sang Jun; Melis, Anastasios; Kim, Jin-Soo; Jin, EonSeon; Bae, Sangsu

    2016-01-01

    Microalgae are versatile organisms capable of converting CO2, H2O, and sunlight into fuel and chemicals for domestic and industrial consumption. Thus, genetic modifications of microalgae for enhancing photosynthetic productivity, and biomass and bio-products generation are crucial for both academic and industrial applications. However, targeted mutagenesis in microalgae with CRISPR-Cas9 is limited. Here we report, a one-step transformation of Chlamydomonas reinhardtii by the DNA-free CRISPR-Cas9 method rather than plasmids that encode Cas9 and guide RNAs. Outcome was the sequential CpFTSY and ZEP two-gene knockout and the generation of a strain constitutively producing zeaxanthin and showing improved photosynthetic productivity. PMID:27466170

  20. DNA-free two-gene knockout in Chlamydomonas reinhardtii via CRISPR-Cas9 ribonucleoproteins

    Science.gov (United States)

    Baek, Kwangryul; Kim, Duk Hyoung; Jeong, Jooyeon; Sim, Sang Jun; Melis, Anastasios; Kim, Jin-Soo; Jin, EonSeon; Bae, Sangsu

    2016-01-01

    Microalgae are versatile organisms capable of converting CO2, H2O, and sunlight into fuel and chemicals for domestic and industrial consumption. Thus, genetic modifications of microalgae for enhancing photosynthetic productivity, and biomass and bio-products generation are crucial for both academic and industrial applications. However, targeted mutagenesis in microalgae with CRISPR-Cas9 is limited. Here we report, a one-step transformation of Chlamydomonas reinhardtii by the DNA-free CRISPR-Cas9 method rather than plasmids that encode Cas9 and guide RNAs. Outcome was the sequential CpFTSY and ZEP two-gene knockout and the generation of a strain constitutively producing zeaxanthin and showing improved photosynthetic productivity. PMID:27466170

  1. Improved generation of rat gene knockouts by target-selected mutagenesis in mismatch repair-deficient animals.

    NARCIS (Netherlands)

    van Boxtel, R.; Toonen, P.W.; Verheul, M.; van Roekel, H.S.; Nijman, I.J.; Guryev, V.; Cuppen, E.

    2008-01-01

    BACKGROUND: The laboratory rat (Rattus norvegicus) is one of the preferred model organisms in physiological and pharmacological research, although the availability of specific genetic models, especially gene knockouts, is limited. N-ethyl-N-nitrosourea (ENU)-driven target-selected mutagenesis is cur

  2. Polymorphisms in DNA Repair Genes (APEX1, XPD, XRCC1 and XRCC3 and Risk of Preeclampsia in a Mexican Mestizo Population

    Directory of Open Access Journals (Sweden)

    Ada Sandoval-Carrillo

    2014-03-01

    Full Text Available Variations in genes involved in DNA repair systems have been proposed as risk factors for the development of preeclampsia (PE. We conducted a case-control study to investigate the association of Human apurinic/apyrimidinic (AP endonuclease (APEX1 Asp148Glu (rs1130409, Xeroderma Pigmentosum group D (XPD Lys751Gln (rs13181, X-ray repair cross-complementing group 1 (XRCC Arg399Gln (rs25487 and X-ray repair cross-complementing group 3 (XRCC3 Thr241Met (rs861539 polymorphisms with PE in a Mexican population. Samples of 202 cases and 350 controls were genotyped using RTPCR. Association analyses based on a χ2 test and binary logistic regression were performed to determine the odds ratio (OR and a 95% confidence interval (95% CI for each polymorphism. The allelic frequencies of APEX1 Asp148Glu polymorphism showed statistical significant differences between preeclamptic and normal women (p = 0.036. Although neither of the polymorphisms proved to be a risk factor for the disease, the APEX1 Asp148Glu polymorphism showed a tendency of association (OR: 1.74, 95% CI = 0.96–3.14 and a significant trend (p for trend = 0.048. A subgroup analyses revealed differences in the allelic frequencies of APEX1 Asp148Glu polymorphism between women with mild preeclampsia and severe preeclampsia (p = 0.035. In conclusion, our results reveal no association between XPD Lys751Gln, XRCC Arg399Gln and XRCC3 Thr241Met polymorphisms and the risk of PE in a Mexican mestizo population; however, the results in the APEX1 Asp148Glu polymorphism suggest the need for future studies using a larger sample size.

  3. GRK5-Knockout Mice Generated by TALEN-Mediated Gene Targeting.

    Science.gov (United States)

    Nanjidsuren, Tsevelmaa; Park, Chae-Won; Sim, Bo-Woong; Kim, Sun-Uk; Chang, Kyu-Tae; Kang, Myung-Hwa; Min, Kwan-Sik

    2016-10-01

    Transcription activator-like effector nucleases (TALENs) are a new type of engineered nuclease that is very effective for directed gene disruption in any genome sequence. We investigated the generation of mice with genetic knockout (KO) of the G protein-coupled receptor kinase (GRK) 5 gene by microinjection of TALEN mRNA. TALEN vectors were designed to target exons 1, 3, and 5 of the mouse GRK5 gene. Flow cytometry showed that the activity of the TALEN mRNAs targeted to exons 1, 3, and 5 was 8.7%, 9.7%, and 12.7%, respectively. The TALEN mRNA for exon 5 was injected into the cytoplasm of 180 one-cell embryos. Of the 53 newborns, three (5.6%) were mutant founders (F0) with mutations. Two clones from F028 showed a 45-bp deletion and F039 showed the same biallelic non-frame-shifting 3-bp deletions. Three clones from F041 were shown to possess a combination of frame-shifting 2-bp deletions. All of the mutations were transmitted through the germline but not to all progenies (37.5%, 37.5%, and 57.1% for the F028, F039, and F041 lines, respectively). The homozygote GRK5-KO mice for 28 and 41 lines created on F3 progenies and the homozygous genotype was confirmed by PCR, T7E1 assay and sequencing. PMID:27565865

  4. Benefits of a "vulnerability gene"? A study in serotonin transporter knockout mice.

    Science.gov (United States)

    Kästner, Niklas; Richter, S Helene; Lesch, Klaus-Peter; Schreiber, Rebecca S; Kaiser, Sylvia; Sachser, Norbert

    2015-04-15

    Over the past years, certain "vulnerability genes" have been identified that play a key role in the development of mood and anxiety disorders. In particular, a low-expressing variant of the human serotonin transporter (5-HTT) gene has been described that renders individuals more susceptible to adverse experience and hence to the development of psychiatric diseases. However, some authors have recently argued that lower 5-HTT expression not only increases vulnerability to adverse experiences, but also enhances susceptibility to beneficial experiences, thus promoting phenotypic plasticity. The aim of the present study was to assess the effects of 5-HTT expression on susceptibility to beneficial experience in a hypothesis-driven experimental approach. Using a well-established rodent model for the human polymorphism, male heterozygous 5-HTT knockout (HET) and 5-HTT wildtype (WT) mice were either provided with the beneficial experience of cohabitation with a female (mating experience) or kept as naïve controls in single-housing conditions. Following the experimental treatment, they were tested for their anxiety-like behaviour and exploratory locomotion in three widely used behavioural tests. Interestingly, while cohabitation reduced anxiety-like behaviour and increased exploratory locomotion in the open field test in HET mice, it did not affect WT mice, pointing to a genotype-dependent susceptibility to the beneficial experience. Thus, our results might support the view of the low expressing version of the 5-HTT gene as a "plasticity" rather than a "vulnerability" variant. PMID:25629942

  5. Acid sphingomyelinase gene knockout ameliorates hyperhomocysteinemic glomerular injury in mice lacking cystathionine-β-synthase.

    Directory of Open Access Journals (Sweden)

    Krishna M Boini

    Full Text Available Acid sphingomyelinase (ASM has been implicated in the development of hyperhomocysteinemia (hHcys-induced glomerular oxidative stress and injury. However, it remains unknown whether genetically engineering of ASM gene produces beneficial or detrimental action on hHcys-induced glomerular injury. The present study generated and characterized the mice lacking cystathionine β-synthase (Cbs and Asm mouse gene by cross breeding Cbs(+/- and Asm(+/- mice. Given that the homozygotes of Cbs(-/-/Asm(-/- mice could not survive for 3 weeks. Cbs(+/-/Asm(+/+, Cbs(+/-/Asm(+/- and Cbs(+/-/Asm(-/- as well as their Cbs wild type littermates were used to study the role of Asm(-/- under a background of Cbs(+/- with hHcys. HPLC analysis revealed that plasma Hcys level was significantly elevated in Cbs heterozygous (Cbs(+/- mice with different copies of Asm gene compared to Cbs(+/+ mice with different Asm gene copies. Cbs(+/-/Asm(+/+ mice had significantly increased renal Asm activity, ceramide production and O(2.(- level compared to Cbs(+/+/Asm(+/+, while Cbs(+/-/Asm(-/- mice showed significantly reduced renal Asm activity, ceramide production and O(2.(- level due to increased plasma Hcys levels. Confocal microscopy demonstrated that colocalization of podocin with ceramide was much lower in Cbs(+/-/Asm(-/- mice compared to Cbs(+/-/Asm(+/+ mice, which was accompanied by a reduced glomerular damage index, albuminuria and proteinuria in Cbs(+/-/Asm(-/- mice. Immunofluorescent analyses of the podocin, nephrin and desmin expression also illustrated less podocyte damages in the glomeruli from Cbs(+/-/Asm(-/- mice compared to Cbs(+/-/Asm(+/+ mice. In in vitro studies of podocytes, hHcys-enhanced O(2.(- production, desmin expression, and ceramide production as well as decreases in VEGF level and podocin expression in podocytes were substantially attenuated by prior treatment with amitriptyline, an Asm inhibitor. In conclusion, Asm gene knockout or corresponding enzyme

  6. Lack of hepcidin gene expression and severe tissue iron overload in upstream stimulatory factor 2 (USF2) knockout mice.

    OpenAIRE

    Nicolas, Gaël; Bennoun, Myriam; Devaux, Isabelle; Beaumont, Carole; Grandchamp, Bernard; Kahn, Axel; Vaulont, Sophie

    2001-01-01

    We previously reported the disruption of the murine gene encoding the transcription factor USF2 and its consequences on glucose-dependent gene regulation in the liver. We report here a peculiar phenotype of Usf2(-/-) mice that progressively develop multivisceral iron overload; plasma iron overcomes transferrin binding capacity, and nontransferrin-bound iron accumulates in various tissues including pancreas and heart. In contrast, the splenic iron content is strikingly lower in knockout animal...

  7. Site-Directed Genome Knockout in Chicken Cell Line and Embryos Can Use CRISPR/Cas Gene Editing Technology

    Directory of Open Access Journals (Sweden)

    Qisheng Zuo

    2016-06-01

    Full Text Available The present study established an efficient genome editing approach for the construction of stable transgenic cell lines of the domestic chicken (Gallus gallus domesticus. Our objectives were to facilitate the breeding of high-yield, high-quality chicken strains, and to investigate gene function in chicken stem cells. Three guide RNA (gRNAs were designed to knockout the C2EIP gene, and knockout efficiency was evaluated in DF-1 chicken fibroblasts and chicken ESCs using the luciferase single-strand annealing (SSA recombination assay, T7 endonuclease I (T7EI assay, and TA clone sequencing. In addition, the polyethylenimine-encapsulated Cas9/gRNA plasmid was injected into fresh fertilized eggs. At 4.5 d later, frozen sections of the embryos were prepared, and knockout efficiency was evaluated by the T7EI assay. SSA assay results showed that luciferase activity of the vector expressing gRNA-3 was double that of the control. Results of the T7EI assay and TA clone sequencing indicated that Cas9/gRNA vector-mediated gene knockdown efficiency was approximately 27% in both DF-1 cells and ESCs. The CRISPR/Cas9 vector was also expressed in chicken embryos, resulting in gene knockdown in three of the 20 embryos (gene knockdown efficiency 15%. Taken together, our results indicate that the CRISPR/Cas9 system can mediate stable gene knockdown at the cell and embryo levels in domestic chickens.

  8. Resources for methylome analysis suitable for gene knockout studies of potential epigenome modifiers

    Directory of Open Access Journals (Sweden)

    Wilson Gareth A

    2012-07-01

    Full Text Available Abstract Background Methylated DNA immunoprecipitation (MeDIP is a popular enrichment based method and can be combined with sequencing (termed MeDIP-seq to interrogate the methylation status of cytosines across entire genomes. However, quality control and analysis of MeDIP-seq data have remained to be a challenge. Results We report genome-wide DNA methylation profiles of wild type (wt and mutant mouse cells, comprising 3 biological replicates of Thymine DNA glycosylase (Tdg knockout (KO embryonic stem cells (ESCs, in vitro differentiated neural precursor cells (NPCs and embryonic fibroblasts (MEFs. The resulting 18 methylomes were analysed with MeDUSA (Methylated DNA Utility for Sequence Analysis, a novel MeDIP-seq computational analysis pipeline for the identification of differentially methylated regions (DMRs. The observed increase of hypermethylation in MEF promoter-associated CpG islands supports a previously proposed role for Tdg in the protection of regulatory regions from epigenetic silencing. Further analysis of genes and regions associated with the DMRs by gene ontology, pathway, and ChIP analyses revealed further insights into Tdg function, including an association of TDG with low-methylated distal regulatory regions. Conclusions We demonstrate that MeDUSA is able to detect both large-scale changes between cells from different stages of differentiation and also small but significant changes between the methylomes of cells that only differ in the KO of a single gene. These changes were validated utilising publicly available datasets and confirm TDG's function in the protection of regulatory regions from epigenetic silencing.

  9. Age-Dependent Defects of Regulatory B Cells in Wiskott-Aldrich Syndrome Gene Knockout Mice.

    Directory of Open Access Journals (Sweden)

    Tadafumi Yokoyama

    Full Text Available The Wiskott-Aldrich syndrome (WAS is a rare X-linked primary immunodeficiency characterized by recurrent infections, thrombocytopenia, eczema, and high incidence of malignancy and autoimmunity. The cellular mechanisms underlying autoimmune complications in WAS have been extensively studied; however, they remain incompletely defined. We investigated the characteristics of IL-10-producing CD19+CD1dhighCD5+ B cells (CD1dhighCD5+ Breg obtained from Was gene knockout (WKO mice and found that their numbers were significantly lower in these mice compared to wild type (WT controls. Moreover, we found a significant age-dependent reduction of the percentage of IL-10-expressing cells in WKO CD1dhighCD5+ Breg cells as compared to age-matched WT control mice. CD1dhighCD5+ Breg cells from older WKO mice did not suppress the in vitro production of inflammatory cytokines from activated CD4+ T cells. Interestingly, CD1dhighCD5+ Breg cells from older WKO mice displayed a basal activated phenotype which may prevent normal cellular responses, among which is the expression of IL-10. These defects may contribute to the susceptibility to autoimmunity with age in patients with WAS.

  10. Conditional knockout of the Slc5a6 gene in mouse intestine impairs biotin absorption.

    Science.gov (United States)

    Ghosal, Abhisek; Lambrecht, Nils; Subramanya, Sandeep B; Kapadia, Rubina; Said, Hamid M

    2013-01-01

    The Slc5a6 gene expresses a plasma membrane protein involved in the transport of the water-soluble vitamin biotin; the transporter is commonly referred to as the sodium-dependent multivitamin transporter (SMVT) because it also transports pantothenic acid and lipoic acid. The relative contribution of the SMVT system toward carrier-mediated biotin uptake in the native intestine in vivo has not been established. We used a Cre/lox technology to generate an intestine-specific (conditional) SMVT knockout (KO) mouse model to address this issue. The KO mice exhibited absence of expression of SMVT in the intestine compared with sex-matched littermates as well as the expected normal SMVT expression in other tissues. About two-thirds of the KO mice died prematurely between the age of 6 and 10 wk. Growth retardation, decreased bone density, decreased bone length, and decreased biotin status were observed in the KO mice. Microscopic analysis showed histological abnormalities in the small bowel (shortened villi, dysplasia) and cecum (chronic active inflammation, dysplasia) of the KO mice. In vivo (and in vitro) transport studies showed complete inhibition in carrier-mediated biotin uptake in the intestine of the KO mice compared with their control littermates. These studies provide the first in vivo confirmation in native intestine that SMVT is solely responsible for intestinal biotin uptake. These studies also provide evidence for a casual association between SMVT function and normal intestinal health.

  11. Effects of activation of central nervous histamine receptors in cardiovascular regulation; studies in H1 and H2 receptor gene knockout mice

    OpenAIRE

    Suzuki, Hideaki; Mobarakeh, Jalal Izadi; Nunoki, Kazuo; Sukegawa, Jun; Watanabe, Haruo; Kuramasu, Atsuo; Watanabe, Takeshi; Yanai, Kazuhiko; Yanagisawa, Teruyuki

    2006-01-01

    To elucidate the central roles of histamine receptors in cardiovascular regulatory system, systolic, mean, and diastolic blood pressures (BPs) and heart rate (HR) were examined in conscious H-1 receptor gene knockout (H1KO) mice, H-2 receptor gene knockout (H2KO) mice, H-1 and H-2 receptor gene double knockout (DKO) mice, and their respective control mice by the tail-cuff system. Histamine, histamine-trifluoromethyl-toluidine derivative (HTMT, an H-1 agonist), dimaprit (an H-2 agonist), and i...

  12. Interleukin-6 gene knockout antagonizes high-fat-induced trabecular bone loss.

    Science.gov (United States)

    Wang, Chunyu; Tian, Li; Zhang, Kun; Chen, Yaxi; Chen, Xiang; Xie, Ying; Zhao, Qian; Yu, Xijie

    2016-10-01

    The purpose of the study was to determine the roles of interleukin-6 (IL6) in fat and bone communication. Male wild-type (WT) mice and IL6 knockout (IL6(-/-)) mice were fed with either regular diet (RD) or high-fat diet (HFD) for 12 weeks. Bone mass and bone microstructure were evaluated by micro-computed tomography. Gene expression related to lipid and bone metabolisms was assayed with real-time quantitative polymerase chain reaction. Bone marrow cells from both genotypes were induced to differentiate into osteoblasts or osteoclasts, and treated with palmitic acid (PA). HFD increased the body weight and fat pad weight, and impaired lipid metabolism in both WT and IL6(-/-) mice. The dysregulation of lipid metabolism was more serious in IL6(-/-) mice. Trabecular bone volume fraction, trabecular bone number and trabecular bone thickness were significantly downregulated in WT mice after HFD than those in the RD (P alkaline phosphatase (ALP) activity and higher mRNA levels of Runx2 and Colla1 than those in WT osteoblasts both in the control and PA treatment group (P < 0.05). IL6(-/-) mice showed significantly lower mRNA levels of PPARγ and leptin and higher mRNA levels of adiponectin in comparison with WT mice on HFD. In conclusion, these findings suggested that IL6 gene deficiency antagonized HFD-induced bone loss. IL6 might bridge lipid and bone metabolisms and could be a new potential therapeutic target for lipid metabolism disturbance-related bone loss. PMID:27493246

  13. High-throughput knockout screen in Schizosaccharomyces pombe identifies a novel gene required for efficient homolog disjunction during meiosis I

    OpenAIRE

    Rumpf, Cornelia; Cipak, Lubos; Novatchkova, Maria; LI, ZHANG; Polakova, Silvia; Dudas, Andrej; Kovacikova, Ines; Miadokova, Eva; Ammerer, Gustav; Gregan, Juraj

    2010-01-01

    Meiosis is the process which produces haploid gametes from diploid precursor cells. This reduction of chromosome number is achieved by two successive divisions. Whereas homologs segregate during meiosis I, sister chromatids segregate during meiosis II. To identify novel proteins required for proper segregation of chromosomes during meiosis, we applied a high-throughput knockout technique to delete 87 S. pombe genes whose expression is upregulated during meiosis and analyzed the mutant phenoty...

  14. Establishment of liver specific glucokinase gene knockout mice:a new animal model for screening anti-diabetic drugs

    Institute of Scientific and Technical Information of China (English)

    Ya-li ZHANG; Xiao-hong TAN; Mei-fang XIAO; Hui LI; Yi-qing Mao; Xiao YANG; Huan-ran TAN

    2004-01-01

    AIM: To characterize the liver-specific role of glucokinase in maintaining glucose homeostasis and to create an animal model for diabetes. METHODS: We performed hepatocyte-specific gene knockout of glucokinase in mice using Cre-loxP gene targeting strategy. First, two directly repeated loxP sequences were inserted to flank the exon 9 and exon 10 of glucokinase in genomic DNA. To achieve this, linearized targeting vector was electroporated into ES cells. Then G418- and Gancyclovir-double-resistant clones were picked and screened by PCR analysis and the positives identified by PCR were confirmed by Southern blot. A targeted clone was selected for microinjection into C57BL/6J blastocysts and implanted into pseudopregnant FVB recipient. Chimeric mice and their offspring were analyzed by Southern blot. Then by intercrossing the Alb-Cre transgenic mice with mice containing a conditional gk allele, we obtained mice with liver-specific glucokinase gene knockout. RESULTS: Among 161 double resistant clones 4 were positive to PCR and Southern blot and only one was used for further experiments. Eventually we generated the liver specific glucokinase knockout mice. These mice showed increased glucose level with age and at the age of 6 weeks fasting blood glucose level was significantly higher than control and they also displayed impaired glucose tolerance. CONCLUSION: Our studies indicate that hepatic glucokinase plays an important role in glucose homeostasis and its deficiencies contribute to the development of diabetes. The liver glucokinase knockout mouse is an ideal animal model for MODY2, and it also can be applied for screening anti-diabetic drugs.

  15. Different Effects of sgRNA Length on CRISPR-mediated Gene Knockout Efficiency.

    Science.gov (United States)

    Zhang, Jian-Ping; Li, Xiao-Lan; Neises, Amanda; Chen, Wanqiu; Hu, Lin-Ping; Ji, Guang-Zhen; Yu, Jun-Yao; Xu, Jing; Yuan, Wei-Ping; Cheng, Tao; Zhang, Xiao-Bing

    2016-01-01

    CRISPR-Cas9 is a powerful genome editing technology, yet with off-target effects. Truncated sgRNAs (17nt) have been found to decrease off-target cleavage without affecting on-target disruption in 293T cells. However, the potency of 17nt sgRNAs relative to the full-length 20nt sgRNAs in stem cells, such as human mesenchymal stem cells (MSCs) and induced pluripotent stem cells (iPSCs), has not been assessed. Using a GFP reporter system, we found that both 17nt and 20nt sgRNAs expressed by lentiviral vectors induce ~95% knockout (KO) in 293T cells, whereas the KO efficiencies are significantly lower in iPSCs (60-70%) and MSCs (65-75%). Furthermore, we observed a decrease of 10-20 percentage points in KO efficiency with 17nt sgRNAs compared to full-length sgRNAs in both iPSCs and MSCs. Off-target cleavage was observed in 17nt sgRNAs with 1-2nt but not 3-4nt mismatches; whereas 20nt sgRNAs with up to 5nt mismatches can still induce off-target mutations. Of interest, we occasionally observed off-target effects induced by the 17nt but not the 20nt sgRNAs. These results indicate the importance of balancing on-target gene cleavage potency with off-target effects: when efficacy is a major concern such as genome editing in stem cells, the use of 20nt sgRNAs is preferable. PMID:27338021

  16. Molecular characterization and development of Sarcocystis speeri sarcocysts in gamma interferon gene knockout mice.

    Science.gov (United States)

    Dubey, J P; Verma, S K; Dunams, D; Calero-Bernal, R; Rosenthal, B M

    2015-11-01

    The North American opossum (Didelphis virginiana) is the definitive host for at least three named species of Sarcocystis: Sarcocystis falcatula, Sarcocystis neurona and Sarcocystis speeri. The South American opossums (Didelphis albiventris, Didelphis marsupialis and Didelphis aurita) are definitive hosts for S. falcatula and S. lindsayi. The sporocysts of these Sarcocystis species are similar morphologically. They are also not easily distinguished genetically because of the difficulties of DNA extraction from sporocysts and availability of distinguishing genetic markers. Some of these species can be distinguished by bioassay; S. neurona and S. speeri are infective to gamma interferon gene knockout (KO) mice, but not to budgerigars (Melopsittacus undulatus); whereas S. falcatula and S. lindsayi are infective to budgerigars but not to KO mice. The natural intermediate host of S. speeri is unknown. In the present study, development of sarcocysts of S. speeri in the KO mice is described. Sarcocysts were first seen at 12 days post-inoculation (p.i.), and they became macroscopic (up to 4 mm long) by 25 days p.i. The structure of the sarcocyst wall did not change from the time bradyzoites had formed at 50-220 days p.i. Sarcocysts contained unique villar protrusions, 'type 38'. The polymerase chain reaction amplifications and sequences analysis of three nuclear loci (18S rRNA, 28S rRNA and ITS1) and two mitochondrial loci (cox1 and cytb) of S. speeri isolate from an Argentinean opossum (D. albiventris) confirmed its membership among species of Sarcocystis and indicated an especially close relationship to another parasite in this genus that employs opossums as its definitive host, S. neurona. These results should be useful in finding natural intermediate host of S. speeri. PMID:26303093

  17. Model-driven in Silico glpC Gene Knockout Predicts Increased Succinate Production from Glycerol in Escherichia Coli

    Directory of Open Access Journals (Sweden)

    Bashir Sajo Mienda

    2015-04-01

    Full Text Available Metabolic engineered targeting for increased succinate production in Escherichia coli using glycerol as a low cost carbon source has attracted global attention in recent years. Succinate production in engineered E. coli has progressed significantly using an experimental trial and error approach. The use of a model-guided, targeted metabolic gene knockout prediction for increased succinate production from glycerol under anaerobic conditions in E. coli still remains largely underexplored. In this study, we applied a model-driven, targeted glpC/b2243 in silico metabolic gene knockout using E. coli genome scale model iJO1366 under the OptFlux software platform with the aim of predicting high succinate flux. The results indicated that the mutant model lacking the glpC/b2243 gene will demonstrate increased succinate flux that is 30% higher than its wild-type control model. We can hypothesize that an additional NADH molecule was generated following the deletion of the gene and/or the alternatively preferred GldA-DhaKLM fermentative route for glycerol metabolism in E. coli may have been activated. Although the exact metabolic mechanism involved in increasing the succinate flux still remains obscure; the current study informs other studies that a model-driven, metabolic glpC/b2243 gene knockout could be applicable in filling our knowledge gap using a comprehensive experimental inquiry in the future; leading to a better understanding of the underlying metabolic function of this gene in relation to succinate production in E. coli from glycerol.

  18. True-breeding targeted gene knock-out in barley using designer TALE-nuclease in haploid cells.

    Directory of Open Access Journals (Sweden)

    Maia Gurushidze

    Full Text Available Transcription activator-like effector nucleases (TALENs are customizable fusion proteins able to cleave virtually any genomic DNA sequence of choice, and thereby to generate site-directed genetic modifications in a wide range of cells and organisms. In the present study, we expressed TALENs in pollen-derived, regenerable cells to establish the generation of instantly true-breeding mutant plants. A gfp-specific TALEN pair was expressed via Agrobacterium-mediated transformation in embryogenic pollen of transgenic barley harboring a functional copy of gfp. Thanks to the haploid nature of the target cells, knock-out mutations were readily detected, and homozygous primary mutant plants obtained following genome duplication. In all, 22% of the TALEN transgenics proved knocked out with respect to gfp, and the loss of function could be ascribed to the deletions of between four and 36 nucleotides in length. The altered gfp alleles were transmitted normally through meiosis, and the knock-out phenotype was consistently shown by the offspring of two independent mutants. Thus, here we describe the efficient production of TALEN-mediated gene knock-outs in barley that are instantaneously homozygous and non-chimeric in regard to the site-directed mutations induced. This TALEN approach has broad applicability for both elucidating gene function and tailoring the phenotype of barley and other crop species.

  19. Effects of Chronic Mild Stress in Female Bax Inhibitor-1-Gene Knockout Mice

    OpenAIRE

    Sui, Zhi-Yan; Chae, Han-Jung; Huang, Guang-Biao; Zhao, Tong; Shrestha Muna, Sushma; Chung, Young-Chul

    2012-01-01

    Objective The anti-apoptotic protein Bax inhibitor-1 (BI-1) is a regulator of apoptosis linked to endoplasmic reticulum (ER) stress, and BI-1-/- mice exhibit increased sensitivity to tissue damage. The purpose of this study was to investigate the role of BI-1 in the pathogenesis of chronic mild stress (CMS)-induced depression-like behaviors in BI-1-/- mice. Methods We delivered CMS for 2 or 6 weeks in BI-1-knockout and wild-type mice. Control groups of BI-1-knockout and wild-type mice were le...

  20. Novel therapeutic targets in osteoarthritis: Narrative review on knock-out genes involved in disease development in mouse animal models.

    Science.gov (United States)

    Veronesi, Francesca; Della Bella, Elena; Cepollaro, Simona; Brogini, Silvia; Martini, Lucia; Fini, Milena

    2016-05-01

    Osteoarthritis (OA) can affect every joint, especially the knee. Given the complexity of this pathology, OA is difficult to treat with current therapies, which only relieve pain and inflammation and are not capable of restoring tissues once OA has started. Currently, researchers focus on finding a therapeutic strategy that may help to arrest disease progression. The present narrative review gives an overview of the genes involved in the development and progression of OA, assessing in vivo studies performed in knock-out mice affected by OA, to suggest new therapeutic strategies. The article search was performed on the PubMed database and www.webofknowledge.com website with the following keywords: "knee osteoarthritis" AND "knockout mice". The included studies were in English and published from 2005 to 2015. Additional papers were found within the references of the selected articles. In the 55 analyzed in vivo studies, genes mainly affected chondrocyte homeostasis, inflammatory processes, extracellular matrix and the relationship between obesity and OA. Genes are defined as inducing, preventing and not influencing OA. This review shows that joint homeostasis depends on a variety of genetic factors, and preventing or restoring the loss of a gene encoding for protective proteins, or inhibiting the expression of proteins that induce OA, might be a potential therapeutic approach. However, conclusions cannot be drawn because of the wide variability concerning the technique used for OA induction, the role of the genes, the method for tissue evaluations and the lack of assessments of all joint tissues. PMID:27059198

  1. 小鼠基因敲除的研究进展%Advances of gene knockout in mice

    Institute of Scientific and Technical Information of China (English)

    张剑; 杨晓梅; 高建刚

    2011-01-01

    With the successful completion of the Human Genome Project (HGP), biological research of the post-genome era has a urgent need for an effective gene function analysis. Application of the knockout mouse model provides a strong support for the study of gene function and the search for new therapeutic interventions in human disease. Two different techniques, gene targeting and gene trapping produce knockout mice from embryonic stem cells (ES cells). The characteristic of gene trapping is high-throughput, random, and sequence tagged, while gene targeting is a specific gene knockout. Two decades ago, the first gene targeting and gene trapping mice were generated. In recent years, new tools for gene targeting and gene trapping have been emerging, and the related organizations have been formed. These organi- zations can knock out genes in the mouse genome using these two methods. The international gene trap consortium (IGTC) and the knockout mouse project (KOMP) have begun to create a worldwide resource for research facilities, and plan to knock out all the mouse genes. KOMP organizers consider it as important as the HGP. From conventional gene targeting to high throughput conditional gene targeting, gene targeting methods have changed. The combined advantages of trapping and targeting enhance the gene trapping spectrum and gene targeting efficiency. As a newly developed insertional mutation system, transposons in trapping genes have more advantages than retrovirus. Emergence of the international knockout mouse consortium (IKMC) is the beginning of global cooperation. The organization is committed to systematically knock out all genes in the mouse genorne, and then to carry out functional genomics studies.%随着人类基因组计划(HGP)的顺利完成,后基因时代的生物学研究迫切需要一种有效的基因功能分析方法。基因敲除小鼠模型的应用,为研究基因的功能和寻找新的治疗人类疾病的干预措施

  2. New insight into the role of the β3 subunit of the GABAA-R in development, behavior, body weight regulation, and anesthesia revealed by conditional gene knockout

    Directory of Open Access Journals (Sweden)

    Hileman Stanley M

    2007-10-01

    Full Text Available Abstract Background The β3 subunit of the γ-aminobutyric acid type A receptor (GABAA-R has been reported to be important for palate formation, anesthetic action, and normal nervous system function. This subunit has also been implicated in the pathogenesis of Angelman syndrome and autism spectrum disorder. To further investigate involvement of this subunit, we previously produced mice with a global knockout of β3. However, developmental abnormalities, compensation, reduced viability, and numerous behavioral abnormalities limited the usefulness of that murine model. To overcome many of these limitations, a mouse line with a conditionally inactivated β3 gene was engineered. Results Gene targeting and embryonic stem cell technologies were used to create mice in which exon 3 of the β3 subunit was flanked by loxP sites (i.e., floxed. Crossing the floxed β3 mice to a cre general deleter mouse line reproduced the phenotype of the previously described global knockout. Pan-neuronal knockout of β3 was achieved by crossing floxed β3 mice to Synapsin I-cre transgenic mice. Palate development was normal in pan-neuronal β3 knockouts but ~61% died as neonates. Survivors were overtly normal, fertile, and were less sensitive to etomidate. Forebrain selective knockout of β3 was achieved using α CamKII-cre transgenic mice. Palate development was normal in forebrain selective β3 knockout mice. These knockouts survived the neonatal period, but ~30% died between 15–25 days of age. Survivors had reduced reproductive fitness, reduced sensitivity to etomidate, were hyperactive, and some became obese. Conclusion Conditional inactivation of the β3 gene revealed novel insight into the function of this GABAA-R subunit. The floxed β3 knockout mice described here will be very useful for conditional knockout studies to further investigate the role of the β3 subunit in development, ethanol and anesthetic action, normal physiology, and pathophysiologic processes.

  3. Learning Apex programming

    CERN Document Server

    Kaufman, Matt

    2015-01-01

    If you are a developer who has some object-oriented programming experience, Learning Apex Programming is the perfect book for you. This book is most appropriate for developers who wish to gain an understanding of the Force.com platform and how to use Apex to create business applications.

  4. Investigation on the Metabolic Regulation of pgi gene knockout Escherichia coli by Enzyme Activities and Intracellular Metabolite Concentrations

    Directory of Open Access Journals (Sweden)

    Nor ‘Aini, A. R.

    2006-01-01

    Full Text Available An integrated analysis of the cell growth characteristics, enzyme activities, intracellular metabolite concentrations was made to investigate the metabolic regulation of pgi gene knockout Escherichia coli based on batch culture and continuous culture which was performed at the dilution rate of 0.2h-1. The enzymatic study identified that pathways of pentose phosphate, ED pathway and glyoxylate shunt were all active in pgi mutant. The glycolysis enzymes i.e glyceraldehyde-3-phosphate dehydrogenase, fructose diphosphatase, pyruvate kinase, triose phosphate isomerase were down regulated implying that the inactivation of pgi gene reduced the carbon flux through glycolytic pathway. Meanwhile, the pentose phosphate pathway was active as a major route for intermediary carbohydrate metabolism instead of glycolysis. The pentose phosphate pathway generates most of the major reducing co-factor NADPH as shown by the increased of NADPH/NADP+ ratio in the mutant when compared with the parent strain. The fermentative enzymes such as acetate kinase and lactate dehydrogenase were down regulated in the mutant. Knockout of pgi gene results in the significant increase in the intracellular concentration of glucose-6-phosphate and decrease in the concentration of oxaloacetate. The slow growth rate of the mutant was assumed to be affected by the accumulation of glucose-6-phosphate and imbalance of NADPH reoxidation.

  5. Lack of hepcidin gene expression and severe tissue iron overload in upstream stimulatory factor 2 (USF2) knockout mice.

    Science.gov (United States)

    Nicolas, G; Bennoun, M; Devaux, I; Beaumont, C; Grandchamp, B; Kahn, A; Vaulont, S

    2001-07-17

    We previously reported the disruption of the murine gene encoding the transcription factor USF2 and its consequences on glucose-dependent gene regulation in the liver. We report here a peculiar phenotype of Usf2(-/-) mice that progressively develop multivisceral iron overload; plasma iron overcomes transferrin binding capacity, and nontransferrin-bound iron accumulates in various tissues including pancreas and heart. In contrast, the splenic iron content is strikingly lower in knockout animals than in controls. To identify genes that may account for the abnormalities of iron homeostasis in Usf2(-/-) mice, we used suppressive subtractive hybridization between livers from Usf2(-/-) and wild-type mice. We isolated a cDNA encoding a peptide, hepcidin (also referred to as LEAP-1, for liver-expressed antimicrobial peptide), that was very recently purified from human blood ultrafiltrate and from urine as a disulfide-bonded peptide exhibiting antimicrobial activity. Accumulation of iron in the liver has been recently reported to up-regulate hepcidin expression, whereas our data clearly show that a complete defect in hepcidin expression is responsible for progressive tissue iron overload. The striking similarity of the alterations in iron metabolism between HFE knockout mice, a murine model of hereditary hemochromatosis, and the Usf2(-/-) hepcidin-deficient mice suggests that hepcidin may function in the same regulatory pathway as HFE. We propose that hepcidin acts as a signaling molecule that is required in conjunction with HFE to regulate both intestinal iron absorption and iron storage in macrophages. PMID:11447267

  6. Claudin-2 knockout by TALEN-mediated gene targeting in MDCK cells: claudin-2 independently determines the leaky property of tight junctions in MDCK cells.

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    Shinsaku Tokuda

    Full Text Available Tight junctions (TJs regulate the movements of substances through the paracellular pathway, and claudins are major determinants of TJ permeability. Claudin-2 forms high conductive cation pores in TJs. The suppression of claudin-2 expression by RNA interference in Madin-Darby canine kidney (MDCK II cells (a low-resistance strain of MDCK cells was shown to induce a three-fold increase in transepithelial electrical resistance (TER, which, however, was still lower than in high-resistance strains of MDCK cells. Because RNA interference-mediated knockdown is not complete and only reduces gene function, we considered the possibility that the remaining claudin-2 expression in the knockdown study caused the lower TER in claudin-2 knockdown cells. Therefore, we investigated the effects of claudin-2 knockout in MDCK II cells by establishing claudin-2 knockout clones using transcription activator-like effector nucleases (TALENs, a recently developed genome editing method for gene knockout. Surprisingly, claudin-2 knockout increased TER by more than 50-fold in MDCK II cells, and TER values in these cells (3000-4000 Ω·cm2 were comparable to those in the high-resistance strains of MDCK cells. Claudin-2 re-expression restored the TER of claudin-2 knockout cells dependent upon claudin-2 protein levels. In addition, we investigated the localization of claudin-1, -2, -3, -4, and -7 at TJs between control MDCK cells and their respective knockout cells using their TALENs. Claudin-2 and -7 were less efficiently localized at TJs between control and their knockout cells. Our results indicate that claudin-2 independently determines the 'leaky' property of TJs in MDCK II cells and suggest the importance of knockout analysis in cultured cells.

  7. Pathogenicty and immune prophylaxis of cag pathogenicity island gene knockout homogenic mutants

    Institute of Scientific and Technical Information of China (English)

    Huan-Jian Lin; Jing Xue; Yang Bai; Ji-De Wang; Ya-Li Zhang; Dian-Yuan Zhou

    2004-01-01

    AIM: To clarify the role of cag pathogenicity island (cagPAI)of Helicobacter pylori(H pylori) in the pathogenicity and immune prophylaxis of H pyloriinfection.METHODS: Three pairs of H pylori including 3 strains of cagPAI positive wildtype bacteria and their cagPAI knockout homogenic mutants were utilized. H pylori binding to the gastric epithelial cells was analyzed by flow cytometry assays.Apoptosis of gastric epithelial cells induced by H pylori was determined by ELISA assay. Prophylaxis effect of the wildtype and mutant strains was compared by immunization with the sonicate of the bacteria into mice model.RESULTS: No difference was found in the apoptasis between cagPAI positive and knockout H pylori strains in respective of the ability in the binding to gastric epithelial cells as well as the induction of apoptosis. Both types of the bacteria were able to protect the mice from the infection of H pylori after immunization, with no difference between them regarding to the protection rate as well as the stimulation of the proliferation of splenocytes of the mice.CONCLUSION: The role of cagPAI in the pathogenicity and prophylaxis of H pylori infection remains to be cleared.

  8. A gene regulatory network model for floral transition of the shoot apex in maize and its dynamic modeling.

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    Zhanshan Dong

    Full Text Available The transition from the vegetative to reproductive development is a critical event in the plant life cycle. The accurate prediction of flowering time in elite germplasm is important for decisions in maize breeding programs and best agronomic practices. The understanding of the genetic control of flowering time in maize has significantly advanced in the past decade. Through comparative genomics, mutant analysis, genetic analysis and QTL cloning, and transgenic approaches, more than 30 flowering time candidate genes in maize have been revealed and the relationships among these genes have been partially uncovered. Based on the knowledge of the flowering time candidate genes, a conceptual gene regulatory network model for the genetic control of flowering time in maize is proposed. To demonstrate the potential of the proposed gene regulatory network model, a first attempt was made to develop a dynamic gene network model to predict flowering time of maize genotypes varying for specific genes. The dynamic gene network model is composed of four genes and was built on the basis of gene expression dynamics of the two late flowering id1 and dlf1 mutants, the early flowering landrace Gaspe Flint and the temperate inbred B73. The model was evaluated against the phenotypic data of the id1 dlf1 double mutant and the ZMM4 overexpressed transgenic lines. The model provides a working example that leverages knowledge from model organisms for the utilization of maize genomic information to predict a whole plant trait phenotype, flowering time, of maize genotypes.

  9. Highly efficient gene knockout by injection of TALEN mRNAs into oocytes and host transfer in Xenopus laevis

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    Keisuke Nakajima

    2015-01-01

    Full Text Available Zinc-finger nucleases, transcription activator-like effector nucleases (TALENs and the CRISPR/Cas (clustered regularly interspaced short palindromic repeats/CRISPR-associated proteins system are potentially powerful tools for producing tailor-made knockout animals. However, their mutagenic activity is not high enough to induce mutations at all loci of a target gene throughout an entire tadpole. In this study, we present a highly efficient method for introducing gene modifications at almost all target sequences in randomly selected embryos. The gene modification activity of TALEN is enhanced by adopting the host-transfer technique. In our method, the efficiency is further improved by injecting TALEN mRNAs fused to the 3′UTR of the Xenopus DEADSouth gene into oocytes, which are then transferred into a host female frog, where they are ovulated and fertilized. The addition of the 3′UTR of the DEADSouth gene promotes mRNA translation in the oocytes and increases the expression of TALEN proteins to near-maximal levels three hours post fertilization (hpf. In contrast, TALEN mRNAs without this 3′UTR are translated infrequently in oocytes. Our data suggest that genomic DNA is more sensitive to TALEN proteins from fertilization to the midblastula (MBT stage. Our method works by increasing the levels of TALEN proteins during the pre-MBT stages.

  10. Gamma aminobutyric acid transporter subtype 1 gene knockout mice: a new model for attention deficit/hyperactivity disorder

    Institute of Scientific and Technical Information of China (English)

    Ping Yang; Guoqiang Cai; Youqing Cai; Jian Fei; Guoxiang Liu

    2013-01-01

    Attention deficit/hyperactivity disorder (ADHD) is characterized by hyperactivity,impaired sustained attention,impulsivity,and is usually accompanied by varying degrees of learning difficulties and lack of motor coordination.However,the pathophysiology and etiology of ADHD remain inconclusive so far.Our previous studies have demonstrated that the gamma aminobutyric acid transporter subtype 1 (GAT1) gene knockout (ko) mouse (gat1-/-)is hyperactive and exhibited impaired memory performance in the Morris water maze.In the current study,we found that the gat1-/-mice showed low levels of attentional focusing and increased impulsivity.In addition,the gat1-/-mice displayed ataxia characterized by defects in motor coordination and balance skills.The hyperactivity in the ko mice was reduced by both methylphenidate and amphetamine.Collectively,these results suggest that GAT1 ko mouse is a new animal model for ADHD studying and GAT1 may be a new target to treat ADHD.

  11. Rapid-throughput skeletal phenotyping of 100 knockout mice identifies 9 new genes that determine bone strength.

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    J H Duncan Bassett

    Full Text Available Osteoporosis is a common polygenic disease and global healthcare priority but its genetic basis remains largely unknown. We report a high-throughput multi-parameter phenotype screen to identify functionally significant skeletal phenotypes in mice generated by the Wellcome Trust Sanger Institute Mouse Genetics Project and discover novel genes that may be involved in the pathogenesis of osteoporosis. The integrated use of primary phenotype data with quantitative x-ray microradiography, micro-computed tomography, statistical approaches and biomechanical testing in 100 unselected knockout mouse strains identified nine new genetic determinants of bone mass and strength. These nine new genes include five whose deletion results in low bone mass and four whose deletion results in high bone mass. None of the nine genes have been implicated previously in skeletal disorders and detailed analysis of the biomechanical consequences of their deletion revealed a novel functional classification of bone structure and strength. The organ-specific and disease-focused strategy described in this study can be applied to any biological system or tractable polygenic disease, thus providing a general basis to define gene function in a system-specific manner. Application of the approach to diseases affecting other physiological systems will help to realize the full potential of the International Mouse Phenotyping Consortium.

  12. Knockout and functional analysis of two DExD/H-box family helicase genes in Sulfolobus islandicus REY15A.

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    Song, Xueguo; Huang, Qihong; Ni, Jinfeng; Yu, Yang; Shen, Yulong

    2016-07-01

    DExD/H-box helicases represent the largest family of helicases. They belong to superfamily 2 helicases and participate in nucleotide metabolism, ribosome biogenesis, and nucleocytoplasmic transport. The biochemical properties and structures of some DExD/H-box helicases in the archaea have been documented, but many of them have not been characterized; and reports on in vivo functional analyses are limited. In this study, we attempted gene knockout of 8 putative DExD/H-box helicases in Sulfolobus islandicus REY15A and obtained two deletion mutants, SiRe_0681 and SiRe_1605. We determined that ΔSiRe_0681 grew faster than wild type cells in the presence of methyl methanesulfonate (MMS). Flow cytometry analysis showed that this strain had fewer G1/S phase cells than the wild type, and the genes coding for cell division proteins were up-regulated. The stain ΔSiRe_1605 was more sensitive to MMS than the wild type cell, and many nucleotide metabolism and DNA repair enzymes were found to be down-regulated. Intriguingly, deletion of either gene led to silencing simultaneously of over 80 genes located at a specific region. This study provides a novel insight into the in vivo functions of predicted DExD/H-box family helicases in the archaea. PMID:27290726

  13. Model-guided metabolic gene knockout of gnd for enhanced succinate production in Escherichia coli from glucose and glycerol substrates.

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    Mienda, Bashir Sajo; Shamsir, Mohd Shahir; Illias, Rosli Md

    2016-04-01

    The metabolic role of 6-phosphogluconate dehydrogenase (gnd) under anaerobic conditions with respect to succinate production in Escherichia coli remained largely unspecified. Herein we report what are to our knowledge the first metabolic gene knockout of gnd to have increased succinic acid production using both glucose and glycerol substrates in E. coli. Guided by a genome scale metabolic model, we engineered the E. coli host metabolism to enhance anaerobic production of succinic acid by deleting the gnd gene, considering its location in the boundary of oxidative and non-oxidative pentose phosphate pathway. This strategy induced either the activation of malic enzyme, causing up-regulation of phosphoenolpyruvate carboxylase (ppc) and down regulation of phosphoenolpyruvate carboxykinase (ppck) and/or prevents the decarboxylation of 6 phosphogluconate to increase the pool of glyceraldehyde-3-phosphate (GAP) that is required for the formation of phosphoenolpyruvate (PEP). This approach produced a mutant strain BMS2 with succinic acid production titers of 0.35 g l(-1) and 1.40 g l(-1) from glucose and glycerol substrates respectively. This work further clearly elucidates and informs other studies that the gnd gene, is a novel deletion target for increasing succinate production in E. coli under anaerobic condition using glucose and glycerol carbon sources. The knowledge gained in this study would help in E. coli and other microbial strains development for increasing succinate production and/or other industrial chemicals. PMID:26878126

  14. Upregulated Expression of Cytotoxicity-Related Genes in IFN-γ Knockout Mice with Schistosoma japonicum Infection

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    Xiaotang Du

    2011-01-01

    Full Text Available It is well accepted that IFN-γ is important to the development of acquired resistance against murine schistosomiasis. However, the in vivo role of this immunoregulatory cytokine in helminth infection needs to be further investigated. In this study, parasite burden and host immune response were observed in IFN-γ knockout mice (IFNg KO infected with Schistosoma japonicum for 6 weeks. The results suggested that deficiency in IFN-γ led to decreased egg burden in mice, with low schistosome-specific IgG antibody response and enhanced activation of T cells during acute infection. Microarray and qRT-PCR data analyses showed significant upregulation of some cytotoxicity-related genes, including those from the granzyme family, tumor necrosis factor, Fas Ligand, and chemokines, in the spleen cells of IFNg KO mice. Furthermore, CD8+ cells instead of NK cells of IFNg KO mice exhibited increased transcription of cytotoxic genes compared with WT mice. Additionally, Schistosoma japonicum-specific egg antigen immunization also could activate CD8+ T cells to upregulate the expression of cytotoxic genes in IFNg KO mice. Our data suggest that IFN-γ is not always a positive regulator of immune responses. In certain situations, the disruption of IFN-γ signaling may up-regulate the cytotoxic T-cell-mediated immune responses to the parasite.

  15. Targeted Chromosomal Translocations and Essential Gene Knockout Using CRISPR/Cas9 Technology in Caenorhabditis elegans.

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    Chen, Xiangyang; Li, Mu; Feng, Xuezhu; Guang, Shouhong

    2015-12-01

    Many genes play essential roles in development and fertility; their disruption leads to growth arrest or sterility. Genetic balancers have been widely used to study essential genes in many organisms. However, it is technically challenging and laborious to generate and maintain the loss-of-function mutations of essential genes. The CRISPR/Cas9 technology has been successfully applied for gene editing and chromosome engineering. Here, we have developed a method to induce chromosomal translocations and produce genetic balancers using the CRISPR/Cas9 technology and have applied this approach to edit essential genes in Caenorhabditis elegans. The co-injection of dual small guide RNA targeting genes on different chromosomes resulted in reciprocal translocation between nonhomologous chromosomes. These animals with chromosomal translocations were subsequently crossed with animals that contain normal sets of chromosomes. The F1 progeny were subjected to a second round of Cas9-mediated gene editing. Through this method, we successfully produced nematode strains with specified chromosomal translocations and generated a number of loss-of-function alleles of two essential genes (csr-1 and mes-6). Therefore, our method provides an easy and efficient approach to generate and maintain loss-of-function alleles of essential genes with detailed genetic background information. PMID:26482793

  16. [Constructing recombinant plasmid pSH-CUP and knockout of acid trehalase gene in baker's yeast].

    Science.gov (United States)

    He, Dongqin; Xiao, Dongguang; Lv, Ye

    2008-02-01

    The ATH1 gene encoded acid trehalase in Saccharomyces cerevisiae. The gene disruption cassette combined the heterologous dominant kan(r) resistance marker with a Cre/loxP-mediated marker removal procedure. The gene disruption cassette was produced by PCR using the same long oligonucleotides comprising 50 nucleotides that annealed to sites upstream or downstream of the genomic target sequence to be deleted. After transformation of the linear disruption cassettes with a Cre/loxP-mediated marker into the cells of Saccharomyces cerevisiae BY-6, selected transformants were checked by PCR for correct the integration of the cassette and concurrent deletion of the chromosomal target sequence. The copper-resistance gene (CUP1-MT1) was cloned into pSH47, which yielded pSH-CUP. The recombinant plasmid pSH-CUP was transformed into the cells of Saccharomyces cerevisiae BY-6(delta ATH1, G418(r)), and transformants were selected for copper resistance. Upon expression of the Cre recombinase results in removal of the kan(r) gene, leaving behind a single loxP site at the chromosomal locus. Construction of the recombinant plasmid pSH-CUP avoided inserting non-yeast gene and made the loxP - kanMX - loxP gene disruption cassette more conventional for eukaryotic organism gene disruption.

  17. Development of a transformation system for gene knock-out in the flavinogenic yeast Pichia guilliermondii.

    Science.gov (United States)

    Boretsky, Yuriy R; Pynyaha, Yuriy V; Boretsky, Volodymyr Y; Kutsyaba, Vasyl I; Protchenko, Olga V; Philpott, Caroline C; Sibirny, Andriy A

    2007-07-01

    Pichia guilliermondii is a representative of a yeast species, all of which over-synthesize riboflavin in response to iron deprivation. Molecular genetic studies in this yeast species have been hampered by a lack of strain-specific tools for gene manipulation. Stable P. guilliermondii ura3 mutants were selected on the basis of 5'-fluoroorotic acid resistance. Plasmid carrying Saccharomyces cerevisiae URA3 gene transformed the mutant strains to prototrophy with a low efficiency. Substitution of a single leucine codon CUG by another leucine codon CUC in the URA3 gene increased the efficiency of transformation 100 fold. Deletion cassettes for the RIB1 and RIB7 genes, coding for GTP cyclohydrolase and riboflavin synthase, respectively, were constructed using the modified URA3 gene and subsequently introduced into a P. guilliermondii ura3 strain. Site-specific integrants were identified by selection for the Rib(-) Ura(+) phenotype and confirmed by PCR analysis. Transformation of the P. guilliermondii ura3 strain was performed using electroporation, spheroplasting or lithium acetate treatment. Only the lithium acetate transformation procedure provided selection of uracil prototrophic, riboflavin deficient recombinant strains. Depending on the type of cassette, efficiency of site-specific integration was 0.1% and 3-12% in the case of the RIB1 and RIB7 genes, respectively. We suggest that the presence of the ARS element adjacent to the 3' end of the RIB1 gene significantly reduced the frequency of homologous recombination. Efficient gene deletion in P. guilliermondii can be achieved using the modified URA3 gene of S. cerevisiae flanked by 0.8-0.9 kb sequences homologous to the target gene. PMID:17467833

  18. The role of nuclear factor E2-Related factor 2 and uncoupling protein 2 in glutathione metabolism: Evidence from an in vivo gene knockout study.

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    Chen, Yanyan; Xu, Yuanyuan; Zheng, Hongzhi; Fu, Jingqi; Hou, Yongyong; Wang, Huihui; Zhang, Qiang; Yamamoto, Masayuki; Pi, Jingbo

    2016-09-01

    Nuclear factor E2-related factor 2 (NRF2) and uncoupling protein 2 (UCP2) are indicated to protect from oxidative stress. They also play roles in the homeostasis of glutathione. However, the detailed mechanisms are not well understood. In the present study, we found Nrf2-knockout (Nrf2-KO) mice exhibited altered glutathione homeostasis and reduced expression of various genes involved in GSH biosynthesis, regeneration, utilization and transport in the liver. Ucp2-knockout (Ucp2-KO) mice exhibited altered glutathione homeostasis in the liver, spleen and blood, as well as increased transcript of cystic fibrosis transmembrane conductance regulator in the liver, a protein capable of mediating glutathione efflux. Nrf2-Ucp2-double knockout (DKO) mice showed characteristics of both Nrf2-KO and Ucp2-KO mice. But no significant difference was observed in DKO mice when compared with Nrf2-KO or Ucp2-KO mice, except in blood glutathione levels. These data suggest that ablation of Nrf2 and Ucp2 leads to disrupted GSH balance, which could result from altered expression of genes involved in GSH metabolism. DKO may not evoke more severe oxidative stress than the single gene knockout. PMID:27453341

  19. Whole body analysis of the knockout gene mouse model for cystic fibrosis using thermal and fast neutron activation analysis

    International Nuclear Information System (INIS)

    A genetically engineered 'knockout gene' mouse model for human cystic fibrosis (CF) has been utilized to study bone mineralization. In CF, the so-called cystic fibrosis transmembrane conductance regulator (CFTR) protein, a chloride ion channel, is either absent or defective. To produce the animal model the murine CFTR gene has been inactivated producing CF symptoms in the homozygotic progeny. CF results in abnormal intestinal absorption of minerals and nutrients which presumably results in substandard bone mineralization. The objective of this study was to determine the feasibility of using whole-body thermal and fast neutron activation analysis to determine mineral and trace-element differences between homozygote controls (+/+) and CF (-/-), murine siblings. Gender-matched juvenile +/+ and -/- litter mates were lyophilized and placed in a BN capsule to reduce thermal-neutron activation and irradiated for 10 seconds at φfast ∼ 1 x 1013 n x cm-2 x s-1 using the MURR pneumatic-tube facility. Phosphorus was measured via the 31P15(n,α)28Al13 reaction. After several days decay, the whole-body specimens were re-irradiated in the same facility, but without thermal-neutron shielding, for 5 seconds and the gamma-ray spectrum was recorded at two different decay periods allowing measurement of 77mSe, 24Na, 27mg, 38Cl, 42k, 49Ca, 56Mn, 66Cu and 80Br from the corresponding radiative-capture reactions. (author)

  20. Knockout of the TauT gene predisposes C57BL/6 mice to streptozotocin-induced diabetic nephropathy.

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    Xiaobin Han

    Full Text Available Diabetic nephropathy is the leading cause of end stage renal disease in the world. Although tremendous efforts have been made, scientists have yet to identify an ideal animal model that can reproduce the characteristics of human diabetic nephropathy. In this study, we hypothesize that taurine insufficiency is a critical risk factor for development of diabetic nephropathy associated with diabetes mellitus. This hypothesis was tested in vivo in TauT heterozygous (TauT+/- and homozygous (TauT-/- knockout in C57BL/6 background mice. We have shown that alteration of the TauT gene (also known as SLC6A6 has a substantial effect on the susceptibility to development of extensive diabetic kidney disease in both TauT+/- and TauT-/-mouse models of diabetes. These animals developed histological changes characteristic of human diabetic nephropathy that included glomerulosclerosis, nodular lesions, arteriosclerosis, arteriolar dilation, and tubulointerstitial fibrosis. Immunohistochemical staining of molecular markers of smooth muscle actin, CD34, Ki67 and collagen IV further confirmed these observations. Our results demonstrated that both homozygous and heterozygous TauT gene deletion predispose C57BL/6 mice to develop end-stage diabetic kidney disease, which closely replicates the pathological features of diabetic nephropathy in human diabetic patients.

  1. Identification of 9 uterine genes that are regulated during mouse pregnancy and exhibit abnormal levels in the cyclooxygenase-1 knockout mouse

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    Soper Jessica

    2007-07-01

    Full Text Available Abstract Background Preterm birth is the leading cause of all infant mortality. In 2004, 12.5% of all births were preterm. In order to understand preterm labor, we must first understand normal labor. Since many of the myometrial changes that occur during pregnancy are similar in mice and humans and mouse gestation is short, we have studied the uterine genes that change in the mouse during pregnancy. Here, we used microarray analysis to identify uterine genes in the gravid mouse that are differentially regulated in the cyclooxygenase-1 knockout mouse model of delayed parturition. Methods Gestational d18.0 uteri (n = 4 were collected from pregnant wild-type and cyclooxygenase-1 knockout mice. Part of the uterus was used for frozen sections and RNA was isolated from the remainder. Microarray analysis was performed at the Indiana University School of Medicine Genomic Core and analyzed using the Microarray Data Portal. Northern analysis was performed to confirm microarray data and the genes localized in the gravid uterus by in situ hybridization. Results We identified 277 genes that are abnormally expressed in the gravid d18.0 cyclooxygenase-1 knockout mouse. Nine of these genes are also regulated in the normal murine uterus during the last half of gestation. Many of these genes are involved in the immune response, consistent with an important role of the immune system in parturition. Expression of 4 of these genes; arginase I, IgJ, Tnfrsf9 and troponin; was confirmed by Northern analysis to be mis-regulated during pregnancy in the knockout mouse. In situ hybridization of these genes demonstrated a similar location in the gravid wild-type and Cox-1 knockout mouse uteri. Conclusion To our knowledge, this is the first work to demonstrate the uterine location of these 4 genes in the mouse during late pregnancy. There are several putative transcription factor binding sites that are shared by many of the 9 genes identified here including; estrogen and

  2. Engineering Clostridium beijerinckii with the Cbei_4693 gene knockout for enhanced ferulic acid tolerance.

    Science.gov (United States)

    Liu, Jun; Guo, Ting; Shen, Xiaoning; Xu, Jiahui; Wang, Junzhi; Wang, Yanyan; Liu, Dong; Niu, Huanqing; Liang, Lei; Ying, Hanjie

    2016-07-10

    A mutant strain of Clostridium beijerinckii NCIMB 8052, C. beijerinckii M11, which exhibited ferulic acid tolerance up to 0.9g/L, was generated using atmospheric pressure glow discharge and high-throughput screening. Comparative genomic analysis revealed that this strain harbored a mutation of the Cbei_4693 gene, which encodes a hypothetical protein suspected to be an NADPH-dependent FMN reductase. After disrupting the Cbei_4693 gene in C. beijerinckii NCIMB 8052 using the ClosTron group II intron-based gene inactivation system, we obtained the Cbei_4693 gene inactivated mutant strain, C. beijerinckii 4693::int. Compared with C. beijerinckii NCIMB 8052, 6.23g/L of butanol was produced in P2 medium containing 0.5g/L of ferulic acid by 4693::int, and the ferulic acid tolerance was also significantly increased up to 0.8g/L. These data showed, for the first time, that the Cbei_4693 gene plays an important role in regulating ferulic acid tolerance in ABE fermentation by C. beijerinckii. PMID:27164255

  3. Antinociceptive effects of morphine and naloxone in mu-opioid receptor knockout mice transfected with the MORS196A gene

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    Tao Pao-Luh

    2010-04-01

    Full Text Available Abstract Background Opioid analgesics such as morphine and meperidine have been used to control moderate to severe pain for many years. However, these opioids have many side effects, including the development of tolerance and dependence after long-term use, which has limited their clinical use. We previously reported that mutations in the mu-opioid receptors (MOR S196L and S196A rendered them responsive to the opioid antagonist naloxone without altering the agonist phenotype. In MORS196A knock-in mice, naloxone and naltrexone were antinociceptive but did not cause tolerance or physical dependence. In this study we delivery this mutated MOR gene into pain related pathway to confirm the possibility of in vivo transfecting MORS196A gene and using naloxone as a new analgesic agent. Methods The MOR-knockout (MOR-KO mice were used to investigate whether morphine and naloxone could show antinociceptive effects when MORS196A gene was transfected into the spinal cords of MOR-KO mice. Double-stranded adeno-associated virus type 2 (dsAAV2 was used to deliver the MORS196A-enhanced green fluorescence protein (EGFP gene by microinjected the virus into the spinal cord (S2/S3 dorsal horn region. Tail-flick test was used to measure the antinociceptive effect of drugs. Results Morphine (10 mg/kg, s.c. and naloxone (10 mg/kg, s.c. had no antinociceptive effects in MOR-KO mice before gene transfection. However, two or three weeks after the MOR-S196A gene had been injected locally into the spinal cord of MOR-KO mice, significant antinociceptive effects could be induced by naloxone or morphine. On the other hand, only morphine but not naloxone induced significant tolerance after sub-chronic treatment. Conclusion Transfecting the MORS196A gene into the spinal cord and systemically administering naloxone in MOR-KO mice activated the exogenously delivered mutant MOR and provided antinociceptive effect without causing tolerance. Since naloxone will not activate natural

  4. Disruption of seven hypothetical aryl alcohol dehydrogenase genes from Saccharomyces cerevisiae and construction of a multiple knock-out strain.

    Science.gov (United States)

    Delneri, D; Gardner, D C; Bruschi, C V; Oliver, S G

    1999-11-01

    By in silicio analysis, we have discovered that there are seven open reading frames (ORFs) in Saccharomyces cerevisiae whose protein products show a high degree of amino acid sequence similarity to the aryl alcohol dehydrogenase (AAD) of the lignin-degrading fungus Phanerochaete chrysosporium. Yeast cultures grown to stationary phase display a significant aryl alcohol dehydrogenase activity by degrading aromatic aldehydes to the corresponding alcohols. To study the biochemical and the biological role of each of the AAD genes, a series of mutant strains carrying deletion of one or more of the AAD-coding sequences was constructed by PCR-mediated gene replacement, using the readily selectable marker kanMX. The correct targeting of the PCR-generated disruption cassette into the genomic locus was verified by analytical PCR and by pulse-field gel electrophoresis (PFGE) followed by Southern blot analysis. Double, triple and quadruple mutant strains were obtained by classical genetic methods, while the construction of the quintuple, sextuple and septuple mutants was achieved by using the marker URA3 from Kluyveromyces lactis, HIS3 from Schizosaccharomyces pombe and TRP1 from S. cerevisiae. None of the knock-out strains revealed any mutant phenotype when tested for the degradation of aromatic aldehydes using both spectrophotometry and high performance liquid chromatography (HPLC). Specific tests for changes in the ergosterol and phospholipids profiles did not reveal any mutant phenotype and mating and sporulation efficiencies were not affected in the septuple deletant. Compared to the wild-type strain, the septuple deletant showed an increased resistance to the anisaldehyde, but there is a possibility that the nutritional markers used for gene replacement are causing this effect.

  5. Type II Toxoplasma gondii KU80 knockout strains enable functional analysis of genes required for cyst development and latent infection.

    Science.gov (United States)

    Fox, Barbara A; Falla, Alejandra; Rommereim, Leah M; Tomita, Tadakimi; Gigley, Jason P; Mercier, Corinne; Cesbron-Delauw, Marie-France; Weiss, Louis M; Bzik, David J

    2011-09-01

    Type II Toxoplasma gondii KU80 knockouts (Δku80) deficient in nonhomologous end joining were developed to delete the dominant pathway mediating random integration of targeting episomes. Gene targeting frequency in the type II Δku80 Δhxgprt strain measured at the orotate (OPRT) and the uracil (UPRT) phosphoribosyltransferase loci was highly efficient. To assess the potential of the type II Δku80 Δhxgprt strain to examine gene function affecting cyst biology and latent stages of infection, we targeted the deletion of four parasite antigen genes (GRA4, GRA6, ROP7, and tgd057) that encode characterized CD8(+) T cell epitopes that elicit corresponding antigen-specific CD8(+) T cell populations associated with control of infection. Cyst development in these type II mutant strains was not found to be strictly dependent on antigen-specific CD8(+) T cell host responses. In contrast, a significant biological role was revealed for the dense granule proteins GRA4 and GRA6 in cyst development since brain tissue cyst burdens were drastically reduced specifically in mutant strains with GRA4 and/or GRA6 deleted. Complementation of the Δgra4 and Δgra6 mutant strains using a functional allele of the deleted GRA coding region placed under the control of the endogenous UPRT locus was found to significantly restore brain cyst burdens. These results reveal that GRA proteins play a functional role in establishing cyst burdens and latent infection. Collectively, our results suggest that a type II Δku80 Δhxgprt genetic background enables a higher-throughput functional analysis of the parasite genome to reveal fundamental aspects of parasite biology controlling virulence, pathogenesis, and transmission. PMID:21531875

  6. Genetic background can result in a marked or minimal effect of gene knockout (GPR55 and CB2 receptor) in experimental autoimmune encephalomyelitis models of multiple sclerosis.

    Science.gov (United States)

    Sisay, Sofia; Pryce, Gareth; Jackson, Samuel J; Tanner, Carolyn; Ross, Ruth A; Michael, Gregory J; Selwood, David L; Giovannoni, Gavin; Baker, David

    2013-01-01

    Endocannabinoids and some phytocannabinoids bind to CB1 and CB2 cannabinoid receptors, transient receptor potential vanilloid one (TRPV1) receptor and the orphan G protein receptor fifty-five (GPR55). Studies using C57BL/10 and C57BL/6 (Cnr2 (tm1Zim)) CB2 cannabinoid receptor knockout mice have demonstrated an immune-augmenting effect in experimental autoimmune encephalomyelitis (EAE) models of multiple sclerosis. However, other EAE studies in Biozzi ABH mice often failed to show any treatment effect of either CB2 receptor agonism or antagonism on inhibition of T cell autoimmunity. The influence of genetic background on the induction of EAE in endocannabinoid system-related gene knockout mice was examined. It was found that C57BL/6.GPR55 knockout mice developed less severe disease, notably in female mice, following active induction with myelin oligodendrocyte glycoprotein 35-55 peptide. In contrast C57BL/6.CB2 (Cnr2 (Dgen)) receptor knockout mice developed augmented severity of disease consistent with the genetically and pharmacologically-distinct, Cnr2 (tm1Zim) mice. However, when the knockout gene was bred into the ABH mouse background and EAE induced with spinal cord autoantigens the immune-enhancing effect of CB2 receptor deletion was lost. Likewise CB1 receptor and transient receptor potential vanilloid one knockout mice on the ABH background demonstrated no alteration in immune-susceptibility, in terms of disease incidence and severity of EAE, in contrast to that reported in some C57BL/6 mouse studies. Furthermore the immune-modulating influence of GPR55 was marginal on the ABH mouse background. Whilst sedative doses of tetrahydrocannabinol could induce immunosuppression, this was associated with a CB1 receptor rather than a CB2 receptor-mediated effect. These data support the fact that non-psychoactive doses of medicinal cannabis have a marginal influence on the immune response in MS. Importantly, it adds a note of caution for the translational value of some

  7. Genetic background can result in a marked or minimal effect of gene knockout (GPR55 and CB2 receptor in experimental autoimmune encephalomyelitis models of multiple sclerosis.

    Directory of Open Access Journals (Sweden)

    Sofia Sisay

    Full Text Available Endocannabinoids and some phytocannabinoids bind to CB1 and CB2 cannabinoid receptors, transient receptor potential vanilloid one (TRPV1 receptor and the orphan G protein receptor fifty-five (GPR55. Studies using C57BL/10 and C57BL/6 (Cnr2 (tm1Zim CB2 cannabinoid receptor knockout mice have demonstrated an immune-augmenting effect in experimental autoimmune encephalomyelitis (EAE models of multiple sclerosis. However, other EAE studies in Biozzi ABH mice often failed to show any treatment effect of either CB2 receptor agonism or antagonism on inhibition of T cell autoimmunity. The influence of genetic background on the induction of EAE in endocannabinoid system-related gene knockout mice was examined. It was found that C57BL/6.GPR55 knockout mice developed less severe disease, notably in female mice, following active induction with myelin oligodendrocyte glycoprotein 35-55 peptide. In contrast C57BL/6.CB2 (Cnr2 (Dgen receptor knockout mice developed augmented severity of disease consistent with the genetically and pharmacologically-distinct, Cnr2 (tm1Zim mice. However, when the knockout gene was bred into the ABH mouse background and EAE induced with spinal cord autoantigens the immune-enhancing effect of CB2 receptor deletion was lost. Likewise CB1 receptor and transient receptor potential vanilloid one knockout mice on the ABH background demonstrated no alteration in immune-susceptibility, in terms of disease incidence and severity of EAE, in contrast to that reported in some C57BL/6 mouse studies. Furthermore the immune-modulating influence of GPR55 was marginal on the ABH mouse background. Whilst sedative doses of tetrahydrocannabinol could induce immunosuppression, this was associated with a CB1 receptor rather than a CB2 receptor-mediated effect. These data support the fact that non-psychoactive doses of medicinal cannabis have a marginal influence on the immune response in MS. Importantly, it adds a note of caution for the translational

  8. Identification of gene knockout strategies using a hybrid of an ant colony optimization algorithm and flux balance analysis to optimize microbial strains.

    Science.gov (United States)

    Lu, Shi Jing; Salleh, Abdul Hakim Mohamed; Mohamad, Mohd Saberi; Deris, Safaai; Omatu, Sigeru; Yoshioka, Michifumi

    2014-09-28

    Reconstructions of genome-scale metabolic networks from different organisms have become popular in recent years. Metabolic engineering can simulate the reconstruction process to obtain desirable phenotypes. In previous studies, optimization algorithms have been implemented to identify the near-optimal sets of knockout genes for improving metabolite production. However, previous works contained premature convergence and the stop criteria were not clear for each case. Therefore, this study proposes an algorithm that is a hybrid of the ant colony optimization algorithm and flux balance analysis (ACOFBA) to predict near optimal sets of gene knockouts in an effort to maximize growth rates and the production of certain metabolites. Here, we present a case study that uses Baker's yeast, also known as Saccharomyces cerevisiae, as the model organism and target the rate of vanillin production for optimization. The results of this study are the growth rate of the model organism after gene deletion and a list of knockout genes. The ACOFBA algorithm was found to improve the yield of vanillin in terms of growth rate and production compared with the previous algorithms. PMID:25462325

  9. Gene knockout and overexpression analysis revealed the role of N-acetylmuramidase in autolysis of Lactobacillus delbrueckii subsp. bulgaricus ljj-6.

    Directory of Open Access Journals (Sweden)

    Xiao-Yang Pang

    Full Text Available Autolysis of lactic acid bacteria (LAB plays a vital role in dairy processing. During cheese making, autolysis of LAB affects cheese flavor development through release of intracellular enzymes and restricts the proliferation of cells in yogurt fermentation and probiotics production. In order to explore the mechanism of autolysis, the gene for the autolytic enzymes of L. bulgaricus, N-acetylmuramidase (mur, was cloned and sequenced (GenBank accession number: KF157911. Mur gene overexpression and gene knockout vectors were constructed based on pMG76e and pUC19 vectors. Recombinant plasmids were transformed into L. bulgaricus ljj-6 by electroporation, then three engineered strains with pMG76e-mur vector and fifteen engineered strains with pUC19-mur::EryBII were screened. The autolysis of the mur knockout strain was significantly lower and autolysis of the mur overexpressed strain was significantly higher compared with that of the wild type strain ljj-6. This result suggested that the mur gene played an important role in autolysis of L. bulgaricus. On the other hand, autolytic activity in a low degree was still observed in the mur knockout strain, which implied that other enzymes but autolysin encoded by mur were also involved in autolysis of L. bulgaricus.

  10. Gene knockout and overexpression analysis revealed the role of N-acetylmuramidase in autolysis of Lactobacillus delbrueckii subsp. bulgaricus ljj-6.

    Science.gov (United States)

    Pang, Xiao-Yang; Cui, Wen-Ming; Liu, Lu; Zhang, Shu-Wen; Lv, Jia-Ping

    2014-01-01

    Autolysis of lactic acid bacteria (LAB) plays a vital role in dairy processing. During cheese making, autolysis of LAB affects cheese flavor development through release of intracellular enzymes and restricts the proliferation of cells in yogurt fermentation and probiotics production. In order to explore the mechanism of autolysis, the gene for the autolytic enzymes of L. bulgaricus, N-acetylmuramidase (mur), was cloned and sequenced (GenBank accession number: KF157911). Mur gene overexpression and gene knockout vectors were constructed based on pMG76e and pUC19 vectors. Recombinant plasmids were transformed into L. bulgaricus ljj-6 by electroporation, then three engineered strains with pMG76e-mur vector and fifteen engineered strains with pUC19-mur::EryBII were screened. The autolysis of the mur knockout strain was significantly lower and autolysis of the mur overexpressed strain was significantly higher compared with that of the wild type strain ljj-6. This result suggested that the mur gene played an important role in autolysis of L. bulgaricus. On the other hand, autolytic activity in a low degree was still observed in the mur knockout strain, which implied that other enzymes but autolysin encoded by mur were also involved in autolysis of L. bulgaricus.

  11. Triple immunoglobulin gene knockout transchromosomic cattle: bovine lambda cluster deletion and its effect on fully human polyclonal antibody production.

    Directory of Open Access Journals (Sweden)

    Hiroaki Matsushita

    Full Text Available Towards the goal of producing fully human polyclonal antibodies (hpAbs or hIgGs in transchromosomic (Tc cattle, we previously reported that Tc cattle carrying a human artificial chromosome (HAC comprising the entire unrearranged human immunoglobulin (Ig heavy-chain (hIGH, kappa-chain (hIGK, and lambda-chain (hIGL germline loci produced physiological levels of hIgGs when both of the bovine immunoglobulin mu heavy-chains, bIGHM and bIGHML1, were homozygously inactivated (bIGHM-/-, bIGHML1-/-; double knockouts or DKO. However, because endogenous bovine immunoglobulin light chain loci are still intact, the light chains are produced both from the hIGK and hIGL genomic loci on the HAC and from the endogenous bovine kappa-chain (bIGK and lambda-chain (bIGL genomic loci, resulting in the production of fully hIgGs (both Ig heavy-chains and light-chains are of human origin: hIgG/hIgκ or hIgG/hIgλ and chimeric hIgGs (Ig heavy-chains are of human origin while the Ig light-chains are of bovine origin: hIgG/bIgκ or hIgG/bIgλ. To improve fully hIgG production in Tc cattle, we here report the deletion of the entire bIGL joining (J and constant (C gene cluster (bIGLJ1-IGLC1 to bIGLJ5-IGLC5 by employing Cre/loxP mediated site-specific chromosome recombination and the production of triple knockout (bIGHM-/-, bIGHML1-/- and bIGL-/-; TKO Tc cattle. We further demonstrate that bIGL cluster deletion greatly improves fully hIgGs production in the sera of TKO Tc cattle, with 51.3% fully hIgGs (hIgG/hIgκ plus hIgG/hIgλ.

  12. Experimental evolution and gene knockout studies reveal AcrA-mediated isobutanol tolerance in Ralstonia eutropha.

    Science.gov (United States)

    Bernardi, Amanda C; Gai, Claudia S; Lu, Jingnan; Sinskey, Anthony J; Brigham, Christopher J

    2016-07-01

    Isobutanol (IBT) has attracted much attention from researchers as a next generation drop-in biofuel. Ralstonia eutropha is a gram-negative bacterium which naturally produces polyhydroxybutyrate (PHB), and has been reported to produce IBT after metabolic engineering. Similar to other microbes, R. eutropha experiences toxicity from branched-chain alcohols and is unable to grow in the presence of IBT concentrations higher than 0.5% (v v(-1)). Such low tolerance greatly limits the ability of R. eutropha to grow and produce IBT. In order to study toxicity to the cells, IBT-tolerant strains were developed by experimental evolution, revealing that two genes, previously described as being related to IBT tolerance in Escherichia coli (acrA and acrA6), also presented mutations in R. eutropha evolved strains. The effect on the physiology of the cells of in-frame deletions of each of these genes was assessed in wild type and engineered IBT-producing strains in an attempt to reproduce a tolerant phenotype. The mutant strains' ability to tolerate, consume, and produce IBT were also analyzed. Although deletions of acrA6 and acrA did not significantly improve R. eutropha growth in the presence of IBT, these deletions improved cell survival in the presence of high concentrations of IBT in the extracellular milieu. Moreover, an in-frame acrA deletion in an engineered IBT-producing R. eutropha enhanced the strain's ability to produce IBT, which could potentially be associated with enhanced survival at high IBT concentrations. PMID:26811221

  13. Systematic analysis of Zn2Cys6 transcription factors required for development and pathogenicity by high-throughput gene knockout in the rice blast fungus.

    Directory of Open Access Journals (Sweden)

    Jianping Lu

    2014-10-01

    Full Text Available Because of great challenges and workload in deleting genes on a large scale, the functions of most genes in pathogenic fungi are still unclear. In this study, we developed a high-throughput gene knockout system using a novel yeast-Escherichia-Agrobacterium shuttle vector, pKO1B, in the rice blast fungus Magnaporthe oryzae. Using this method, we deleted 104 fungal-specific Zn(2Cys(6 transcription factor (TF genes in M. oryzae. We then analyzed the phenotypes of these mutants with regard to growth, asexual and infection-related development, pathogenesis, and 9 abiotic stresses. The resulting data provide new insights into how this rice pathogen of global significance regulates important traits in the infection cycle through Zn(2Cys(6TF genes. A large variation in biological functions of Zn(2Cys(6TF genes was observed under the conditions tested. Sixty-one of 104 Zn(2Cys(6 TF genes were found to be required for fungal development. In-depth analysis of TF genes revealed that TF genes involved in pathogenicity frequently tend to function in multiple development stages, and disclosed many highly conserved but unidentified functional TF genes of importance in the fungal kingdom. We further found that the virulence-required TF genes GPF1 and CNF2 have similar regulation mechanisms in the gene expression involved in pathogenicity. These experimental validations clearly demonstrated the value of a high-throughput gene knockout system in understanding the biological functions of genes on a genome scale in fungi, and provided a solid foundation for elucidating the gene expression network that regulates the development and pathogenicity of M. oryzae.

  14. Metabolic engineering of Escherichia coli for the production of l-valine based on transcriptome analysis and in silico gene knockout simulation

    OpenAIRE

    Park, Jin Hwan; Lee, Kwang Ho; Kim, Tae Yong; Lee, Sang Yup

    2007-01-01

    The l-valine production strain of Escherichia coli was constructed by rational metabolic engineering and stepwise improvement based on transcriptome analysis and gene knockout simulation of the in silico genome-scale metabolic network. Feedback inhibition of acetohydroxy acid synthase isoenzyme III by l-valine was removed by site-directed mutagenesis, and the native promoter containing the transcriptional attenuator leader regions of the ilvGMEDA and ilvBN operon was replaced with the tac pro...

  15. 15 years of genetic approaches in vivo for addiction research: opioid receptor and peptide gene knockout in mouse models of drug abuse

    OpenAIRE

    Charbogne, Pauline; Kieffer, Brigitte L.; Befort, Katia

    2013-01-01

    The endogenous opioid system is expressed throughout the brain reinforcement circuitry, and plays a major role in reward processing, mood control and the development of addiction. This neuromodulator system is composed of three receptors, mu, delta and kappa, interacting with a family of opioid peptides derived from POMC (β-endorphin), preproenkephalin (pEnk) and preprodynorphin (pDyn) precursors. Knockout mice targeting each gene of the opioid system have been created almost two decades ago....

  16. PLEKHQ1基因敲除小鼠基因型鉴定方法%The method of the identification of the PLEKHQ1 gene knockout mice

    Institute of Scientific and Technical Information of China (English)

    张鹏飞; 张硌; 陆琤; 周晨辰

    2015-01-01

    目的:探讨鉴定PLEKHQ1基因敲除(KO)小鼠基因型的方法。方法:对PLEKHQ1基因敲除杂合子小鼠进行单独饲养及配种繁殖,繁殖后其子代出现野生型、杂合子型及纯合子型3种基因型,提取每只小鼠的基因组DNA,采用聚合酶链反应(PCR)和变性方法进行基因类型鉴定。结果:采用PCR和变性法成功鉴定出PLEKHQ1基因敲除小鼠的基因型。结论:这种无需T7酶切的小鼠基因型鉴定方法可用于PLEKHQ1基因敲除小鼠的基因型鉴定。%Objective:To identify PLEKHQ1 gene knock-out mice.Methods: The PLEKHQ1 gene knock-out heterozygote mice were bred alone and copulated. The offsprings were to have three genotypes: wild genotype, heterozygote genotype and homozygote genotype. Genomic DNA was obtained from each pups and were subjected to PCR and Denature to identify the genotype. Results: The identification of PLEKHQ1 gene knockout mice is successful.Conclusion: The identification method of PLEKHQ1-KO mice without T7 can correct identify PLEKHQ1 gene knockout mice.

  17. Establishment of IL-31 RA Gene Knockout Homozygous Mice Model%IL-31 RA基因敲除小鼠纯合子模型的建立

    Institute of Scientific and Technical Information of China (English)

    江涛; 高婧; 岳欢; 高雅倩; 黄俊琼

    2015-01-01

    目的:建立IL-31RA基因敲除小鼠纯合子模型,为IL-31RA基因相关研究提供动物模型。方法:IL-31RA基因敲除小鼠严格按照SPF级要求的动物饲养标准进行饲养繁殖,采用聚合酶链式反应( PCR)法鉴定子代小鼠的基因型,RT-PCR法鉴定小鼠IL-31RA mRNA的表达,Western blot鉴定IL-31RA蛋白的表达,HE染色观察小鼠重要脏器的形态学变化。结果:PCR法成功检测出子代小鼠的3种基因型,纯合子基因敲除小鼠未检测出IL-31RA mRNA和IL-31RA蛋白的表达,IL-31RA基因敲除小鼠的重要脏器的形态学特征与野生型小鼠比较无明显变化;基因敲除小鼠可成功饲养繁殖,亦可获得较多的基因敲除纯合子小鼠。结论:成功构建了IL-31RA基因敲除小鼠纯合子模型。%Objective:To establish the IL-31RA gene knockout homozygous mice model and lay the foundation for further study on IL-31 gene. Methods:IL-31RA gene knockout mice were bred and re-produced according to the SPF class animal feeding standard. PCR was used to identify the genotype of the offspring,the expression of IL-31RA mRNA was detected by RT-PCR,expression of IL-31RA pro-tein was detected by Western blot,and morphological changes of vital organs were observed by HE staining . Result:Three genotypes of the offspring of IL-31 RA gene knockout mice were successfully i-dentified;expression of IL-31 RA mRNA and IL-31 RA protein was not detected in IL-31 RA gene knockout homozygous mice. Compared with the wild type mice,morphological characteristics of vital organs of had no significant changes in IL-31RA gene knockout homozygous mice. IL-31RA gene knockout mice could be dred and reproduced successfully. Conclusion:The IL-31RA gene knockout homozygous mice model has been successfully established.

  18. [BLG gene knockout and hLF gene knock-in at BLG locus in goat by TALENs].

    Science.gov (United States)

    Song, Shaozheng; Zhu, Mengmin; Yuan, Yuguo; Rong, Yao; Xu, Sheng; Chen, Si; Mei, Junyan; Cheng, Yong

    2016-03-01

    To knock out β-lactoglobulin (BLG) gene and insert human lactoferrin (hLF) coding sequence at BLG locus of goat, the transcription activator-like effector nucleases (TALEN) mediated recombination was used to edit the BLG gene of goat fetal fibroblast, then as donor cells for somatic cell nuclear transfer. We designed a pair of specific plasmid TALEN-3-L/R for goat BLG exon III recognition sites, and BLC14-TK vector containing a negative selection gene HSV-TK, was used for the knock in of hLF gene. TALENs plasmids were transfected into the goat fetal fibroblast cells, and the cells were screened three days by 2 μg/mL puromycin. DNA cleavage activities of cells were verified by PCR amplification and DNA production sequencing. Then, targeting vector BLC14-TK and plasmids TALEN-3-L/R were co-transfected into goat fetal fibroblasts, both 700 μg/mL G418 and 2 μg/mL GCV were simultaneously used to screen G418-resistant cells. Detections of integration and recombination were implemented to obtain cells with hLF gene site-specific integration. We chose targeting cells as donor cells for somatic cell nuclear transfer. The mutagenicity of TALEN-3-L/R was between 25% and 30%. A total of 335 reconstructed embryos with 6 BLG-/hLF+ targeting cell lines were transferred into 16 recipient goats. There were 9 pregnancies confirmed by ultrasound on day 30 to 35 (pregnancy rate of 39.1%), and one of 50-day-old fetus with BLG-/hLF+ was achieved. These results provide the basis for hLF gene knock-in at BLG locus of goat and cultivating transgenic goat of low allergens and rich hLF in the milk. PMID:27349115

  19. Knockout of the tumor necrosis factor α receptor 1 gene can up-regulate erythropoietin receptor during myocardial ischemia-reperfusion injury in mice

    Institute of Scientific and Technical Information of China (English)

    LI Chang-ling; JIANG Jun; FAN You-qi; FU Guo-sheng; WANG Jia-nan; FAN Wei-ming

    2009-01-01

    Background Tumor necrosis factor α receptor 1 (TNFαR1) plays an important role in the signal pathway of apoptosis.The objective of this study was to investigate the effects of TNFaR1 knockout on the up-regulation of erythropoietin receptor (Epo-R) and the coordinated anti-apoptosis functions during myocardial ischemia-reperfusion injury in mice.Methods The ischemia-reperfusion injury model for cardiomyocytes was performed by ligating the left circumflex branch artery of TNFαR1 knockout (P55-/-) C17 B6 mice, as well as wild-type (P55+/+) C17 B6 mice. Triphenyltetrazolium chloride (TTC) staining was performed to observe the damaged area of the heart. TUNEL staining and DNA fragmentation were used to identify apoptosis. Mitochondrial Bcl-2 and Bax as well as expression of Epo-R and its downstream genes (Jak-2, slat-5, Akt, IkB-α, HIF-1α) were measured by Western blotting. The gene knockout mice were assigned into those undergoing the apoptosis surgical model group (KO group), and those subjected to sham operation (Kos group). Similarly, wild-type mice were either exposed to the surgical model (WT group) or subject to a sham operation (WTs group).Results The myocardial damage ratio of the wild-type group after the operation was significantly higher than that of the knockout group, (50.5±6.4)% vs (36.9±6.9)%, P<0.01. Similarly, TUNEL positive ratio of the wild-type group was significantly higher than that of the knockout group, (63.1±5.6)% vs (42.1±4.7)%, P<0.01. The gray value ratios of Epo-R,Jak-2, stat-5, Akt, IkB-α, HIF-1 and mitochondrial Bcl-2 in the KO group were significantly higher than those of the WT group, P<0.05; however, mitochondrial Bax was significantly lower than that of the WT group significantly (P<0.05).Conclusions Using the ischemia-reperfusion injury model in mice, cardiomyocytes of TNFαR1 knockouts exhibited anti-apoptotic characteristics. This information could be used to coordinate the prevention of myocardial apoptosis by up

  20. Type II Toxoplasma gondii KU80 Knockout Strains Enable Functional Analysis of Genes Required for Cyst Development and Latent Infection ▿ †

    OpenAIRE

    Fox, Barbara A.; Falla, Alejandra; Rommereim, Leah M.; Tomita, Tadakimi; Gigley, Jason P.; Mercier, Corinne; Cesbron-Delauw, Marie-France; Louis M Weiss; Bzik, David J.

    2011-01-01

    Type II Toxoplasma gondii KU80 knockouts (Δku80) deficient in nonhomologous end joining were developed to delete the dominant pathway mediating random integration of targeting episomes. Gene targeting frequency in the type II Δku80 Δhxgprt strain measured at the orotate (OPRT) and the uracil (UPRT) phosphoribosyltransferase loci was highly efficient. To assess the potential of the type II Δku80 Δhxgprt strain to examine gene function affecting cyst biology and latent stages of infection, we t...

  1. Regulation of Gene Expression of Catecholamine Biosynthetic Enzymes in Dopamine-β-Hydroxylase- and CRH-Knockout Mice Exposed to Stress

    OpenAIRE

    Richard, Kvetnansky; Olga, Krizanova; Andrej, Tillinger; Sabban Esther, L.; Thomas Steven, A; Lucia, Kubovcakova

    2008-01-01

    Norepinephrine-deficient mice harbor a disruption of the gene for dopamine-β-hydroxylase (DBH-KO). Corticotropin-releasing hormone knockout mice (CRH-KO) have markedly reduced HPA activity. The aim of the present work was to study how deficiency of DBH and CRH would affect tyrosine hydroxylase (TH), DBH, and phenylethanolamine N-methyltransferase (PNMT) gene expression and protein levels in the adrenal medulla (AM) and stellate ganglia (SG) of control and stressed mice. Both in AM and SG, sin...

  2. Behaviour Study of The Ras-GRF1 Gene knockout Mice%Ras-GRF1基因敲除小鼠的行为学研究

    Institute of Scientific and Technical Information of China (English)

    段巍鹤; 郑宏亮; 陆美林; 万家余; 何秀霞

    2015-01-01

    The Ras genes widely exist in nature, Ras-GRF1 proteins in cell signal transduction, cell differentiation and growth play a very important role. Previous researchs have shown that the Ras genes are closely associated with signal-ing pathways and learning and memory function. This research was study to companed the differences between the gene knockout Ras-GRF1 and will rats on learning and memory by the behavior experiments. It was found that Ras-GRF1 knockout mice learning memory ability weak in wild type mice by Morris water maze,etc.%Ras基因广泛存在于自然界中,其控制的Ras-GRF1蛋白在细胞信号传导,在细胞分化与生长过程中起着极其重要的作用。已有研究表明,Ras基因与信号通路和学习记忆功能密切相关。本实验利用行为学研究敲除Ras-GRF1基因的小鼠在学习记忆上与野生老鼠的差异,通过Morris水迷宫等实验发现Ras-GRF1基因敲除小鼠的学习记忆能力弱于野生型小鼠。

  3. Generation and confirmation of NAAG peptidase gene knockout model%NAAG肽酶基因剔除小鼠模型的建立及鉴定

    Institute of Scientific and Technical Information of China (English)

    崔振文; 张明坤; 钟春龙; 吴增宝; 蔡蕾; 匡颖; 王铸钢; 江基尧; 罗其中

    2012-01-01

    目的 建立N-乙酰天冬氨酰谷氨酸(NAAG)肽酶基因剔除小鼠模型,为在体研究NAAG肽酶基因的生物学功能并揭示其在脑损伤后继发性脑损害进程中的所起的作用创造条件.方法 根据小鼠NAAG肽酶基因组的序列,设计基因剔除策略,构建基因剔除载体NAAG-KO-pBR322,以电穿孔方法将基因剔除载体导入胚胎干细胞(ES),应用G418和更昔洛韦进行正负筛选,获得双抗性克隆,聚合酶链式反应(PCR)鉴定并测序获得正确同源重组的ES细胞克隆.结果 同源重组的ES细胞注入小鼠囊胚后获得11只嵌合率>50%嵌合体雄性小鼠,嵌合体小鼠与C57 BL/6J雌鼠交配后获得11只杂合子小鼠,其中雄性7只,雌性4只.在雌、雄杂合子小鼠交配的后代中获得7只纯合子小鼠,PCR鉴定其基因型,逆转录PCR (RT-PCR)提示该基因鼠未表达NAAG肽酶.结论 我们成功建立了NAAG肽酶基因剔除小鼠模型,其中纯合子小鼠未出现胚胎致死现象;初步的表型观察未发现NAAG肽酶基因剔除小鼠出现异常改变.%Objective To establish N-acetylaspartylglutamate (NAAG) peptidase gene knockout mouse model and to create the condition for farther in vivo study of its biological function and its role in the secondary brain damage after brain injury. Methods According to the NAAG peptidase genomic DNA sequence ,the strategy of gene targeting was established, and the gene knockout vector (NAAG-KO-pBR322) was constructed. Electroporation of embryonic stem (ES) cells with the gene knockout vector and screening of both G418 and Ganciclovir resistant clones were performed. The homologous recombined ES ceD clones were identified by polymerase chain reaction (PCR). Results After transplantation of homologous recombined ES cells into blastocysts through microinjection, there were 11 male chimeras bom with embedment rate >50%. The male chimeras then were bred with C57BL/6J female mice and 7(♂) and 4 (♀) offsprings with

  4. Breeding Reproducing and Identifying for p53 Gene Knockout Mice%p53基因敲除小鼠的饲养繁殖及鉴定

    Institute of Scientific and Technical Information of China (English)

    乔录新; 徐萌; 柴梦音; 乔欣; 陈德喜

    2012-01-01

    目的 为了繁育和鉴定p53基因敲除小鼠,将引进的杂合子小鼠进行饲养繁殖,杂合子用于继续保种.方法 对其幼鼠剪尾提取基因组DNA,采用PCR方法进行基因型鉴定.结果 对引进小鼠已成功饲养和繁殖,并得到纯合基因缺失型小鼠.结论 正确的饲养、繁殖及基因鉴定方法对于基因敲除小鼠的获得和保种具有重要的意义.%Objective To breed and identify p53 gene knockout mice, Heterozygote mice were bred and reproduced. Methods Genome DNA extracted from the mice' s tails were subjected to PCR test for genotype identification. Results Heterozygous were used to acquire baby mice for Protection species. Conclusion The breeding and reproducing were successful and Homozygous genotype mice were acquired. Appropriate methods of feeding, breeding and identifying are important for obtaining gene knockout mice and protecting species.

  5. SETD4基因敲除小鼠的构建及鉴定%Establishment and Identification of SETD4gene knockout mice

    Institute of Scientific and Technical Information of China (English)

    黄穗; 黄梦怡; 钟玙沄; 雷烨铭; 赵舒祺; 蔡军伟; 姜勇; 刘靖华

    2016-01-01

    Objective To study the function of SETD4,the SETD4 gene knockout homozygous mice has been established. Methods SETD4flox/+mice and EIIa-Cre mice were interbred,the offspring of which was genotyping SETD4 +/-.EIIa-Cre were crossed with C57BL/6 mice to obtain the mice with the SETD4+/-genotype,SETD4+/-heterozygous mice were inbred and then the SETD4-/- homozygous mice were gained. PCR was used to identify the genotype of the offspring,the expression of SETD4 mRNA was detected by RT-PCR and qPCR,and morphological changes of liver and lung were observed by HE staining. Result PCR results showed genotypes of the offspring of SETD4 gene knockout mice was in accordance with SETD4-/-. Compared with the wild type mice,expression of SETD4 mRNA in SETD4 gene knockout homozygous mice was significantly decreased,and morphological characteristics of liver and lung in SETD4 gene knockout homozygous mice had no significant changes. Conclusion Wehave successfully generated SETD4 gene knockout homozygous mice which can be used for study ofSETD4 function.%目的:构建并鉴定SETD4基因敲除小鼠,为研究SETD4的生物学功能提供动物模型。方法将引进的SETD4flox/+小鼠与EIIa-Cre小鼠进行杂交繁殖,得到基因型为SETD4+/-.EIIa-Cre的小鼠;再与C57BL/6小鼠杂交去除Cre酶,获得杂合子SETD4+/-小鼠;该小鼠自交获得纯合子SETD4-/-小鼠。通过PCR法鉴定子代小鼠的基因型;RT-PCR、荧光定量PCR方法鉴定纯合子的SETD4基因敲除小鼠SETD4 mRNA表达情况;HE染色观察小鼠肝、肺组织的形态学变化。结果 PCR结果表明子代小鼠的基因型符合SETD4-/-;纯合子基因敲除小鼠SETD4 mRNA水平显著低于野生型小鼠;SETD4基因敲除小鼠肝、肺组织的形态学特征与野生型小鼠相比无明显差异。结论本研究基于Cre/loxp系统,成功构建并鉴定了SETD4基因敲除小鼠。

  6. Cortical Gene Expression After a Conditional Knockout of 67 kDa Glutamic Acid Decarboxylase in Parvalbumin Neurons.

    Science.gov (United States)

    Georgiev, Danko; Yoshihara, Toru; Kawabata, Rika; Matsubara, Takurou; Tsubomoto, Makoto; Minabe, Yoshio; Lewis, David A; Hashimoto, Takanori

    2016-07-01

    In the cortex of subjects with schizophrenia, expression of glutamic acid decarboxylase 67 (GAD67), the enzyme primarily responsible for cortical GABA synthesis, is reduced in the subset of GABA neurons that express parvalbumin (PV). This GAD67 deficit is accompanied by lower cortical levels of other GABA-associated transcripts, including GABA transporter-1, PV, brain-derived neurotrophic factor (BDNF), tropomyosin receptor kinase B, somatostatin, GABAA receptor α1 subunit, and KCNS3 potassium channel subunit mRNAs. In contrast, messenger RNA (mRNA) levels for glutamic acid decarboxylase 65 (GAD65), another enzyme for GABA synthesis, are not altered. We tested the hypothesis that this pattern of GABA-associated transcript levels is secondary to the GAD67 deficit in PV neurons by analyzing cortical levels of these GABA-associated mRNAs in mice with a PV neuron-specific GAD67 knockout. Using in situ hybridization, we found that none of the examined GABA-associated transcripts had lower cortical expression in the knockout mice. In contrast, PV, BDNF, KCNS3, and GAD65 mRNA levels were higher in the homozygous mice. In addition, our behavioral test battery failed to detect a change in sensorimotor gating or working memory, although the homozygous mice exhibited increased spontaneous activities. These findings suggest that reduced GAD67 expression in PV neurons is not an upstream cause of the lower levels of GABA-associated transcripts, or of the characteristic behaviors, in schizophrenia. In PV neuron-specific GAD67 knockout mice, increased levels of PV, BDNF, and KCNS3 mRNAs might be the consequence of increased neuronal activity secondary to lower GABA synthesis, whereas increased GAD65 mRNA might represent a compensatory response to increase GABA synthesis. PMID:26980143

  7. The serotonin transporter knockout rat : A review

    NARCIS (Netherlands)

    Olivier, Jocelien; Cools, Alexander; Ellenbroek, Bart A.; Cuppen, E.; Homberg, Judith; Kalueff, Allan V.; LaPorte, Justin L.

    2010-01-01

    This chapter dicusses the most recent data on the serotonin transporter knock-out rat, a unique rat model that has been generated by target-selected N-ethyl-N-nitrosourea (ENU) driven mutagenesis. The knock-out rat is the result of a premature stopcodon in the serotonin transporter gene, and the abs

  8. Knockout of GH3 genes in the moss Physcomitrella patens leads to increased IAA levels at elevated temperature and in darkness.

    Science.gov (United States)

    Mittag, Jennifer; Gabrielyan, Anastasia; Ludwig-Müller, Jutta

    2015-12-01

    Two proteins of the GRETCHEN HAGEN3 (GH3) family of acyl acid amido synthetases from the moss Physcomitrella patens conjugate indole-3-acetic acid (IAA) to a series of amino acids. The possible function of altered auxin levels in the moss in response to two different growth perturbations, elevated temperatures and darkness, was analyzed using a) the recently described double knockout lines in both P. patens GH3 genes (GH3-doKO) and b) a previously characterized line harboring an auxin-inducible soybean GH3 promoter::reporter fused to β-glucuronidase (G1-GUS). The GUS activity as marker of the auxin response increased at higher temperatures and after cultivation in the darkness for a period of up to four weeks. Generally, the double knockout plants grew more slowly than the wild type (WT). The altered growth conditions influenced the phenotypes of the double knockout lines differently from that of WT moss. Higher temperatures negatively affected GH3-doKO plants compared to WT which was shown by stronger loss of chlorophyll. On the other hand, a positive effect was found on the concentrations of free IAA which increased at 28 °C in the GH3-doKO lines compared to WT plants. A different factor, namely darkness vs. a light/dark cycle caused the adverse phenotype concerning chlorophyll concentrations. Mutant moss plants showed higher chlorophyll concentrations than WT and these correlated with higher free IAA in the plant population that was classified as green. Our data show that growth perturbations result in higher free IAA levels in the GH3-doKO mutants, but in one case - growth in darkness - the mutants could cope better with the condition, whereas at elevated temperatures the mutants were more sensitive than WT. Thus, GH3 function in P. patens WT could lie in the regulation of IAA concentrations under unfavorable environmental conditions.

  9. Mucocele of the petrous apex.

    Science.gov (United States)

    DeLozier, H L; Parkins, C W; Gacek, R R

    1979-02-01

    The first case of a primary mucocele of the petrous apex is presented and the differential diagnosis is briefly discussed. Since this area is unavailable for direct examination, a thorough radiographic evaluation is essential. A mucocele should be suspected when a lytic lesion has a multiloculated appearance and when the contralateral petrous apex is highly pneumatized. Even then, a biopsy may still be needed to make a definitive diagnosis. The appropriate treatment for these cystic lesions is fistulization into a radical mastoid cavity or an exteriorized sphenoid sinus. PMID:429898

  10. Desenvolvimento e avaliação de uma cepa knockout de Brucella abortus obtida pela deleção do gene virB10 Development and evaluation of a strain of Brucella abortus gotten by the knockout of the virB10 gene

    OpenAIRE

    Fabiane G. de Souza; Ana L.A.R. Osório; Bárbara G. Csordas; Rafael Q. Prado; Carina Elisei; Cleber O. Soares; Araújo, Flábio R; Stênio P Fragoso; Grácia M.S. Rosinha

    2009-01-01

    Brucella spp. são bactérias gram-negativas, intracelulares facultativas que são patogênicas para muitas espécies de mamíferos causando a brucelose, uma zoonose difundida mundialmente. Por isso a busca de alternativas de controle mais eficientes se faz necessário como o desenvolvimento de novas cepas que possam ser testadas como potenciais imunógenos. Neste estudo realizou-se a deleção do gene virB10 da cepa S2308 de Brucella abortus gerando uma cepa knockout provavelmente incapaz de produzir ...

  11. The APEX-SZ Instrument

    CERN Document Server

    Schwan, Daniel; Basu, Kaustuv; Bender, Amy N; Bertoldi, Frank; Cho, Hsaio-Mei; Chon, Guyong; Clarke, John; Dobbs, Matt; Ferrusca, Daniel; Gusten, Rolfe; Halverson, Nils W; Holzapfel, William L; Horellou, Cathy; Johansson, Daniel; Johnson, Bradley R; Kennedy, James; Kermish, Zigmund; Kneissl, Ruediger; Lanting, Trevor; Lee, Adrian T; Lueker, Martin; Mehl, Jared; Menten, Karl M; Muders, Dirk; Pacaud, Florian; Plagge, Thomas; Reichardt, Christian L; Richards, Paul L; Schaaf, Rienhold; Schilke, Peter; Sommer, Martin W; Spieler, Helmuth; Tucker, Carole; Weiss, Axel; Westbrook, Benjamin; Zahn, Oliver

    2010-01-01

    The APEX-SZ instrument is a millimeter-wave cryogenic receiver designed to observe galaxy clusters via the Sunyaev-Zel'dovich effect from the 12 m APEX telescope on the Atacama plateau in Chile. The receiver contains a focal plane of 280 superconducting transition-edge sensor (TES) bolometers instrumented with a frequency-domain multiplexed readout system. The bolometers are cooled to 280 mK via a three-stage helium sorption refrigerator and a mechanical pulse-tube cooler. Three warm mirrors, two 4 K lenses, and a horn array couple the TES bolometers to the telescope. APEX-SZ observes in a single frequency band at 150 GHz with 1' angular resolution and a 22' field-of-view, all well suited for cluster mapping. The APEX-SZ receiver has played a key role in the introduction of several new technologies including TES bolometers, the frequency-domain multiplexed readout, and the use of a pulse-tube cooler with bolometers. As a result of these new technologies, the instrument has a higher instantaneous sensitivity a...

  12. Synthetic liver X receptor agonist T0901317 inhibits semicarbazide-sensitive amine oxidase gene expression and activity in apolipoprotein E knockout mice

    Institute of Scientific and Technical Information of China (English)

    Xiaoyan Dai; Xiang Ou; Xinrui Hao; Dongli Cao; Yaling Tang; Yanwei Hu; Xiaoxu Li; Chaoke Tang

    2008-01-01

    Semicarbazide-sensitive amine oxidase(SSAO)catalyzes oxidative deamination of primary aromatic and aliphatic amines.Increased SSAO activity has been found in atherosclerosis and diabetes mellitus.We hypothesize that the anti-atherogenic effect of liver X receptors(LXRs)might be related to the inhibition of SSAD gene expression and its activity.In this study,we investigated the effect of LXR agonist T0901317 on SSAO gene expression and its activity in apolipoprotein E knockout(apoE-/-)mice.Male apoE-/-mice(8 weeks old) were randomly divided into four groups:basal control group;vehicle group;prevention group;and treatment group.SSAO gene expression was analyzed by real-time quantitative polymerase chain reaction and its activity was determined.The activity of superoxide dismutase and content of malondialdehy de in the aorta and liver were also determined.In T0901317-treated mice,SSAO gene expression was significantly decreased in the aorta,liver,small intestine,and brain.SSAO activities in serum and in these tissues were also inhibited.The amount of superoxide dismutase in the aorta and liver of the prevention group and treatment group was significantly higher compared with the vehicle group(P<0.05).Malondialdehyde in the tissues of these two groups was significantly lower compared with the vehicle group(P<0.05).Our results showed that T0901317 inhibits SSAO gene expression and its activity in atherogenic apoE-/-mice.The atheroprotective effect of LXR agonist T0901317 is related to the inhibition of SSAO gene expression and its activity.

  13. Gene Knockout Study Reveals That Cytosolic Ascorbate Peroxidase 2(OsAPX2) Plays a Critical Role in Growth and Reproduction in Rice under Drought, Salt and Cold Stresses

    OpenAIRE

    Zhiguo Zhang; Quian Zhang; Jinxia Wu; Xia Zheng; Sheng Zheng; Xuehui Sun; Quansheng Qiu; Tiegang Lu

    2013-01-01

    Plant ascorbate peroxidases (APXs), enzymes catalyzing the dismutation of H2O2 into H2O and O2, play an important role in reactive oxygen species homeostasis in plants. The rice genome has eight OsAPXs, but their physiological functions remain to be determined. In this report, we studied the function of OsAPX2 gene using a T-DNA knockout mutant under the treatment of drought, salt and cold stresses. The Osapx2 knockout mutant was isolated by a genetic screening of a rice T-DNA insertion libra...

  14. Genetic knockout of the α7 nicotinic acetylcholine receptor gene alters hippocampal long-term potentiation in a background strain-dependent manner.

    Science.gov (United States)

    Freund, Ronald K; Graw, Sharon; Choo, Kevin S; Stevens, Karen E; Leonard, Sherry; Dell'Acqua, Mark L

    2016-08-01

    Reduced α7 nicotinic acetylcholine receptor (nAChR) function is linked to impaired hippocampal-dependent sensory processing and learning and memory in schizophrenia. While knockout of the Chrna7 gene encoding the α7nAChR on a C57/Bl6 background results in changes in cognitive measures, prior studies found little impact on hippocampal synaptic plasticity in these mice. However, schizophrenia is a multi-genic disorder where complex interactions between specific genetic mutations and overall genetic background may play a prominent role in determining phenotypic penetrance. Thus, we compared the consequences of knocking out the α7nAChR on synaptic plasticity in C57/Bl6 and C3H mice, which differ in their basal α7nAChR expression levels. Homozygous α7 deletion in C3H mice, which normally express higher α7nAChR levels, resulted in impaired long-term potentiation (LTP) at hippocampal CA1 synapses, while C3H α7 heterozygous mice maintained robust LTP. In contrast, homozygous α7 deletion in C57 mice, which normally express lower α7nAChR levels, did not alter LTP, as had been previously reported for this strain. Thus, the threshold of Chrna7 expression required for LTP may be different in the two strains. Measurements of auditory gating, a hippocampal-dependent behavioral paradigm used to identify schizophrenia-associated sensory processing deficits, was abnormal in C3H α7 knockout mice confirming that auditory gating also requires α7nAChR expression. Our studies highlight the importance of genetic background on the regulation of synaptic plasticity and could be relevant for understanding genetic and cognitive heterogeneity in human studies of α7nAChR dysfunction in mental disorders.

  15. A Simplified Method for Gene Knockout and Direct Screening of Recombinant Clones for Application in Paenibacillus polymyxa.

    Directory of Open Access Journals (Sweden)

    Seong-Bin Kim

    Full Text Available Paenibacillus polymyxa is a bacterium widely used in agriculture, industry, and environmental remediation because it has multiple functions including nitrogen fixation and produces various biologically active compounds. Among these compounds are the antibiotics polymyxins, and the bacterium is currently being reassessed for medical application. However, a lack of genetic tools for manipulation of P. polymyxa has limited our understanding of the biosynthesis of these compounds.To facilitate an understanding of the genetic determinants of the bacterium, we have developed a system for marker exchange mutagenesis directly on competent cells of P. polymyxa under conditions where homologous recombination is enhanced by denaturation of the suicide plasmid DNA. To test this system, we targeted P. polymyxa α-and β-amylase genes for disruption. Chloramphenicol or erythromycin resistance genes were inserted into the suicide plasmid pGEM7Z-f+ (Promega. To mediate homologous recombination and replacement of the targeted genes with the antibiotic resistance genes nucleotide sequences of the α-and β-amylase genes were cloned into the plasmid flanking the antibiotic resistance genes.We have created a simple system for targeted gene deletion in P. polymyxa E681. We propose that P. polymyxa isogenic mutants could be developed using this system of marker exchange mutagenesis. α-and β-amylase genes provide a useful tool for direct recombinant screening in P. polymyxa.

  16. Breeding and Genotyping of PTA-1-/-/ApoE-/-Double-gene Knockout Mice%PTA-1-/-/ApoE-/-双基因敲除小鼠的繁育及基因型鉴定

    Institute of Scientific and Technical Information of China (English)

    董子龙; 庄然; 张宇丝; 金伯泉; 张圆

    2012-01-01

    目的 建立(PTA-1-/-/ApoE-/-)双基因敲除小鼠(double-gene knockout,DKO)模型,探讨该小鼠的繁育及鉴定方法,为进一步利用该小鼠研究相关疾病奠定基础.方法 将引进的PTA-1-/-及ApoE-/-基因敲除小鼠通过杂交和互交的方法进行繁殖,以得到DKO小鼠.结果 经过PCR基因鉴定的方法证实PTA-1-/-/ApoE-/-双基因敲除小鼠繁育成功.结论 正确的饲养繁殖及鉴定方法是获得该DKO纯合子小鼠的有效途径.%Objective To breed and identify the PTA-1-/-/ApoE-/- double-gene knockout (DKO) mice and to establish an animal model to further study the role of PTA-1 molecule in diseases. Method PTA-1 gene knockout mice were paired with the ApoE gene knockout mice in different ways. Genomic DNA were isolated from the tails and analyzed by PCR. Result Genotyping analysis identified that we established PTA-1 -/-/ApoE-/- DKO mice successfully. Conclusion It is feasible to breed PTA-1 -/-/ApoE-/- DKO mice with the PTA-1 and ApoE gene knockout mice. PCR can be used to identify the genotype of the DKO mice precisely.

  17. Muc2和DCN基因敲除小鼠食子现象的初步研究%Preliminary study of kronismus in Muc2 and DCN gene knockout mice

    Institute of Scientific and Technical Information of China (English)

    李巍; 贺国洋

    2013-01-01

    Objective To explore the differences in kronismus of Muc2 and DCN gene knockout mice. Methods Knockout homozygote males and females ( Muc2 -/ - ,DCN -/ - ) mice were mated respectively according to 1 :1 or 1 ;2 ratio. The average litter size of 1 - 3 generations, parity interval time, and kronismus in Muc2 and DCN gene knockout mice were observed. Results An average litter size of Muc2 gene knockout mice was 5.80 ±0.95. The average birth interval time was 42. 29 ±2. 28 days. The average DCN knockout mice seed production was 3. 85 ±0. 76, and the average birth interval time was 24. 86 ± 10. 42 days. There were significant differences between Muc2 and DCN gene knockout mice in the average litter size, parity interval time, and kronismus. Conclusions The reproductive performance of the two groups of gene knockout mice are different, indicating that Muc2 and DCN genes may be associated with reproductive function.%目的 探讨Muc2和DCN基因敲除小鼠繁殖能力和食子现象的异同.方法 分别将Muc2和DCN基因敲除纯合子雌雄小鼠按1∶1或1∶的合笼,观察1~3胎产仔量、胎次间隔时间、出生存活率和食子现象.结果 Muc2基因敲除小鼠平均产子量5.80±0.95只,平均胎次间隔时间(42.29±2.28) d;DCN基因敲除小鼠平均产子量3.85±0.76只,平均胎次间隔时间(24.86±10.42)d.Muc2和DCN基因敲除小鼠在产仔量、胎次间隔时间、出生存活率和食子率差异均存在显著性.结论 两组基因敲除小鼠繁殖性能有差异,揭示可能与Muc2和DCN基因有关.

  18. The FKBP5 Gene Affects Alcohol Drinking in Knockout Mice and Is Implicated in Alcohol Drinking in Humans

    Science.gov (United States)

    Qiu, Bin; Luczak, Susan E.; Wall, Tamara L.; Kirchhoff, Aaron M.; Xu, Yuxue; Eng, Mimy Y.; Stewart, Robert B.; Shou, Weinian; Boehm, Stephen L.; Chester, Julia A.; Yong, Weidong; Liang, Tiebing

    2016-01-01

    FKBP5 encodes FK506-binding protein 5, a glucocorticoid receptor (GR)-binding protein implicated in various psychiatric disorders and alcohol withdrawal severity. The purpose of this study is to characterize alcohol preference and related phenotypes in Fkbp5 knockout (KO) mice and to examine the role of FKBP5 in human alcohol consumption. The following experiments were performed to characterize Fkpb5 KO mice. (1) Fkbp5 KO and wild-type (WT) EtOH consumption was tested using a two-bottle choice paradigm; (2) The EtOH elimination rate was measured after intraperitoneal (IP) injection of 2.0 g/kg EtOH; (3) Blood alcohol concentration (BAC) was measured after 3 h limited access of alcohol; (4) Brain region expression of Fkbp5 was identified using LacZ staining; (5) Baseline corticosterone (CORT) was assessed. Additionally, two SNPs, rs1360780 (C/T) and rs3800373 (T/G), were selected to study the association of FKBP5 with alcohol consumption in humans. Participants were college students (n = 1162) from 21–26 years of age with Chinese, Korean or Caucasian ethnicity. The results, compared to WT mice, for KO mice exhibited an increase in alcohol consumption that was not due to differences in taste sensitivity or alcohol metabolism. Higher BAC was found in KO mice after 3 h of EtOH access. Fkbp5 was highly expressed in brain regions involved in the regulation of the stress response, such as the hippocampus, amygdala, dorsal raphe and locus coeruleus. Both genotypes exhibited similar basal levels of plasma corticosterone (CORT). Finally, single nucleotide polymorphisms (SNPs) in FKBP5 were found to be associated with alcohol drinking in humans. These results suggest that the association between FKBP5 and alcohol consumption is conserved in both mice and humans. PMID:27527158

  19. The FKBP5 Gene Affects Alcohol Drinking in Knockout Mice and Is Implicated in Alcohol Drinking in Humans

    Directory of Open Access Journals (Sweden)

    Bin Qiu

    2016-08-01

    Full Text Available FKBP5 encodes FK506-binding protein 5, a glucocorticoid receptor (GR-binding protein implicated in various psychiatric disorders and alcohol withdrawal severity. The purpose of this study is to characterize alcohol preference and related phenotypes in Fkbp5 knockout (KO mice and to examine the role of FKBP5 in human alcohol consumption. The following experiments were performed to characterize Fkpb5 KO mice. (1 Fkbp5 KO and wild-type (WT EtOH consumption was tested using a two-bottle choice paradigm; (2 The EtOH elimination rate was measured after intraperitoneal (IP injection of 2.0 g/kg EtOH; (3 Blood alcohol concentration (BAC was measured after 3 h limited access of alcohol; (4 Brain region expression of Fkbp5 was identified using LacZ staining; (5 Baseline corticosterone (CORT was assessed. Additionally, two SNPs, rs1360780 (C/T and rs3800373 (T/G, were selected to study the association of FKBP5 with alcohol consumption in humans. Participants were college students (n = 1162 from 21–26 years of age with Chinese, Korean or Caucasian ethnicity. The results, compared to WT mice, for KO mice exhibited an increase in alcohol consumption that was not due to differences in taste sensitivity or alcohol metabolism. Higher BAC was found in KO mice after 3 h of EtOH access. Fkbp5 was highly expressed in brain regions involved in the regulation of the stress response, such as the hippocampus, amygdala, dorsal raphe and locus coeruleus. Both genotypes exhibited similar basal levels of plasma corticosterone (CORT. Finally, single nucleotide polymorphisms (SNPs in FKBP5 were found to be associated with alcohol drinking in humans. These results suggest that the association between FKBP5 and alcohol consumption is conserved in both mice and humans.

  20. The FKBP5 Gene Affects Alcohol Drinking in Knockout Mice and Is Implicated in Alcohol Drinking in Humans.

    Science.gov (United States)

    Qiu, Bin; Luczak, Susan E; Wall, Tamara L; Kirchhoff, Aaron M; Xu, Yuxue; Eng, Mimy Y; Stewart, Robert B; Shou, Weinian; Boehm, Stephen L; Chester, Julia A; Yong, Weidong; Liang, Tiebing

    2016-08-05

    FKBP5 encodes FK506-binding protein 5, a glucocorticoid receptor (GR)-binding protein implicated in various psychiatric disorders and alcohol withdrawal severity. The purpose of this study is to characterize alcohol preference and related phenotypes in Fkbp5 knockout (KO) mice and to examine the role of FKBP5 in human alcohol consumption. The following experiments were performed to characterize Fkpb5 KO mice. (1) Fkbp5 KO and wild-type (WT) EtOH consumption was tested using a two-bottle choice paradigm; (2) The EtOH elimination rate was measured after intraperitoneal (IP) injection of 2.0 g/kg EtOH; (3) Blood alcohol concentration (BAC) was measured after 3 h limited access of alcohol; (4) Brain region expression of Fkbp5 was identified using LacZ staining; (5) Baseline corticosterone (CORT) was assessed. Additionally, two SNPs, rs1360780 (C/T) and rs3800373 (T/G), were selected to study the association of FKBP5 with alcohol consumption in humans. Participants were college students (n = 1162) from 21-26 years of age with Chinese, Korean or Caucasian ethnicity. The results, compared to WT mice, for KO mice exhibited an increase in alcohol consumption that was not due to differences in taste sensitivity or alcohol metabolism. Higher BAC was found in KO mice after 3 h of EtOH access. Fkbp5 was highly expressed in brain regions involved in the regulation of the stress response, such as the hippocampus, amygdala, dorsal raphe and locus coeruleus. Both genotypes exhibited similar basal levels of plasma corticosterone (CORT). Finally, single nucleotide polymorphisms (SNPs) in FKBP5 were found to be associated with alcohol drinking in humans. These results suggest that the association between FKBP5 and alcohol consumption is conserved in both mice and humans.

  1. The FKBP5 Gene Affects Alcohol Drinking in Knockout Mice and Is Implicated in Alcohol Drinking in Humans.

    Science.gov (United States)

    Qiu, Bin; Luczak, Susan E; Wall, Tamara L; Kirchhoff, Aaron M; Xu, Yuxue; Eng, Mimy Y; Stewart, Robert B; Shou, Weinian; Boehm, Stephen L; Chester, Julia A; Yong, Weidong; Liang, Tiebing

    2016-01-01

    FKBP5 encodes FK506-binding protein 5, a glucocorticoid receptor (GR)-binding protein implicated in various psychiatric disorders and alcohol withdrawal severity. The purpose of this study is to characterize alcohol preference and related phenotypes in Fkbp5 knockout (KO) mice and to examine the role of FKBP5 in human alcohol consumption. The following experiments were performed to characterize Fkpb5 KO mice. (1) Fkbp5 KO and wild-type (WT) EtOH consumption was tested using a two-bottle choice paradigm; (2) The EtOH elimination rate was measured after intraperitoneal (IP) injection of 2.0 g/kg EtOH; (3) Blood alcohol concentration (BAC) was measured after 3 h limited access of alcohol; (4) Brain region expression of Fkbp5 was identified using LacZ staining; (5) Baseline corticosterone (CORT) was assessed. Additionally, two SNPs, rs1360780 (C/T) and rs3800373 (T/G), were selected to study the association of FKBP5 with alcohol consumption in humans. Participants were college students (n = 1162) from 21-26 years of age with Chinese, Korean or Caucasian ethnicity. The results, compared to WT mice, for KO mice exhibited an increase in alcohol consumption that was not due to differences in taste sensitivity or alcohol metabolism. Higher BAC was found in KO mice after 3 h of EtOH access. Fkbp5 was highly expressed in brain regions involved in the regulation of the stress response, such as the hippocampus, amygdala, dorsal raphe and locus coeruleus. Both genotypes exhibited similar basal levels of plasma corticosterone (CORT). Finally, single nucleotide polymorphisms (SNPs) in FKBP5 were found to be associated with alcohol drinking in humans. These results suggest that the association between FKBP5 and alcohol consumption is conserved in both mice and humans. PMID:27527158

  2. APEX Refrigeration Company Limited: Brand Identity Project

    OpenAIRE

    Dee, Peter

    2004-01-01

    APEX Refrigeration offers Refrigeration, Air Conditioning and Catering services across Ireland. Apex Refrigeration required a clean, bold and modern logo using cool colours and imagery. Peter Dee - Strategic Design and Marketing Consultant, was responsible for the design and development of the brand identity for the APEX Refrigeration Company which was used on business cards, letterhead, vehicle livery and e-Commerce website.

  3. Novel and natural knockout lung cancer cell lines for the LKB1/STK11 tumor suppressor gene

    OpenAIRE

    Carretero, J.; Medina, P.P. (Pedro P.); Pio, R. (Rubén); Montuenga, L M; Sanchez-Cespedes, M.

    2004-01-01

    Germline mutations of the LKB1 gene are responsible for Peutz-Jeghers syndrome (PJS), an autosomal dominant inherited disorder bestowing an increased risk of cancer. We have recently demonstrated that LKB1 inactivating mutations are not confined to PJS, but also appear in lung adenocarcinomas of sporadic origin, including primary tumors and lung cancer cell lines. To accurately determine the frequency of inactivating LKB1 gene mutations in lung tumors we have sequenced the complete coding reg...

  4. Generation of Interleukin-2 Receptor Gamma Gene Knockout Pigs from Somatic Cells Genetically Modified by Zinc Finger Nuclease-Encoding mRNA

    Science.gov (United States)

    Watanabe, Masahito; Nakano, Kazuaki; Matsunari, Hitomi; Matsuda, Taisuke; Maehara, Miki; Kanai, Takahiro; Kobayashi, Mirina; Matsumura, Yukina; Sakai, Rieko; Kuramoto, Momoko; Hayashida, Gota; Asano, Yoshinori; Takayanagi, Shuko; Arai, Yoshikazu; Umeyama, Kazuhiro; Nagaya, Masaki; Hanazono, Yutaka; Nagashima, Hiroshi

    2013-01-01

    Zinc finger nuclease (ZFN) is a powerful tool for genome editing. ZFN-encoding plasmid DNA expression systems have been recently employed for the generation of gene knockout (KO) pigs, although one major limitation of this technology is the use of potentially harmful genome-integrating plasmid DNAs. Here we describe a simple, non-integrating strategy for generating KO pigs using ZFN-encoding mRNA. The interleukin-2 receptor gamma (IL2RG) gene was knocked out in porcine fetal fibroblasts using ZFN-encoding mRNAs, and IL2RG KO pigs were subsequently generated using these KO cells through somatic cell nuclear transfer (SCNT). The resulting IL2RG KO pigs completely lacked a thymus and were deficient in T and NK cells, similar to human X-linked SCID patients. Our findings demonstrate that the combination of ZFN-encoding mRNAs and SCNT provides a simple robust method for producing KO pigs without genomic integration. PMID:24130776

  5. A one-step cloning method for the construction of somatic cell gene targeting vectors: application to production of human knockout cell lines

    Directory of Open Access Journals (Sweden)

    Liu Yi

    2012-10-01

    Full Text Available Abstract Background Gene targeting is a powerful method that can be used for examining the functions of genes. Traditionally, the construction of knockout (KO vectors requires an amplification step to obtain two homologous, large fragments of genomic DNA. Restriction enzymes that cut at unique recognitions sites and numerous cloning steps are then carried out; this is often a time-consuming and frustrating process. Results We have developed a one-step cloning method for the insertion of two arms into a KO vector using exonuclease III. We modified an adeno-associated virus KO shuttle vector (pTK-LoxP-NEO-AAV to yield pAAV-LIC, which contained two cassettes at the two multiple-cloning sites. The vector was digested with EcoRV to give two fragments. The two homologous arms, which had an overlap of 16 bases with the ends of the vector fragments, were amplified by polymerase chain reaction. After purification, the four fragments were mixed and treated with exonuclease III, then transformed into Escherichia coli to obtain the desired clones. Using this method, we constructed SirT1 and HDAC2 KO vectors, which were used to establish SirT1 KO cells from the colorectal cancer cell line (HCT116 and HDAC2 KO cells from the colorectal cancer cell line (DLD1. Conclusions Our method is a fast, simple, and efficient technique for cloning, and has great potential for high-throughput construction of KO vectors.

  6. Knockout Zbtb33 gene results in an increased locomotion, exploration and pre-pulse inhibition in mice.

    Science.gov (United States)

    Kulikov, Alexander V; Korostina, Valeria S; Kulikova, Elizabeth A; Fursenko, Dariya V; Akulov, Andrey E; Moshkin, Mikhail P; Prokhortchouk, Egor B

    2016-01-15

    The Zbtb33 gene encodes the Kaiso protein-a bimodal transcriptional repressor. Here, the effects of Zbtb33 gene disruption on the brain and behaviour of the Kaiso-deficient (KO) and C57BL/6 (WT) male mice were investigated. Behaviour was studied using the open field, novel object, elevated plus maze and acoustic startle reflex tests. Brain morphology was investigated with magnetic resonance imaging. Biogenic amine levels and gene expression in the brain were measured with high-performance liquid chromatography and quantitative real-time RT-PCR, respectively. Zbtb33 gene mRNA was not detected in the brain of KO mice. KO mice exhibited increased locomotion, exploration in the open field, novel object and elevated plus-maze test. At the same time, Zbtb33 gene disruption did not alter anxiety-related behaviour in the elevated plus-maze test. KO mice showed elevated amplitudes and pre-pulse inhibitions of the acoustic startle reflex. These behavioural alterations were accompanied by significant reductions in the volumes of the lateral ventricles without significant alterations in the volumes of the hippocampus, striatum, thalamus and corpus callosum. Norepinephrine concentration was reduced in the hypothalami and hippocampi in KO mice, while the levels of serotonin, dopamine, their metabolites as well as mRNA of the gene coding brain-derived neurotrophic factor were not altered in the brain of KO mice compared to WT mice. This study is the first to reveal the involvement of the Zbtb33 gene in the regulation of behaviour and the central nervous system. PMID:26454239

  7. Oracle APEX 4.2 cookbook

    CERN Document Server

    Van Zoest, Michel

    2013-01-01

    As a Cookbook, this book enables you to create APEX web applications and to implement features with immediately usable recipes that unleash the powerful functionality of Oracle APEX 4.2. Each recipe is presented as a separate, standalone entity and the reading of other, prior recipes is not required.It can be seen as a reference and a practical guide to APEX development.This book is aimed both at developers new to the APEX environment and at intermediate developers. More advanced developers will also gain from the information at hand.If you are new to APEX you will find recipes to start develo

  8. Monte Carlo studies of APEX

    Energy Technology Data Exchange (ETDEWEB)

    Ahmad, I.; Back, B.B.; Betts, R.R. [and others

    1995-08-01

    An essential component in the assessment of the significance of the results from APEX is a demonstrated understanding of the acceptance and response of the apparatus. This requires detailed simulations which can be compared to the results of various source and in-beam measurements. These simulations were carried out using the computer codes EGS and GEANT, both specifically designed for this purpose. As far as is possible, all details of the geometry of APEX were included. We compared the results of these simulations with measurements using electron conversion sources, positron sources and pair sources. The overall agreement is quite acceptable and some of the details are still being worked on. The simulation codes were also used to compare the results of measurements of in-beam positron and conversion electrons with expectations based on known physics or other methods. Again, satisfactory agreement is achieved. We are currently working on the simulation of various pair-producing scenarios such as the decay of a neutral object in the mass range 1.5-2.0 MeV and also the emission of internal pairs from nuclear transitions in the colliding ions. These results are essential input to the final results from APEX on cross section limits for various, previously proposed, sharp-line producing scenarios.

  9. Type II Toxoplasma gondii KU80 Knockout Strains Enable Functional Analysis of Genes Required for Cyst Development and Latent Infection ▿ †

    Science.gov (United States)

    Fox, Barbara A.; Falla, Alejandra; Rommereim, Leah M.; Tomita, Tadakimi; Gigley, Jason P.; Mercier, Corinne; Cesbron-Delauw, Marie-France; Weiss, Louis M.; Bzik, David J.

    2011-01-01

    Type II Toxoplasma gondii KU80 knockouts (Δku80) deficient in nonhomologous end joining were developed to delete the dominant pathway mediating random integration of targeting episomes. Gene targeting frequency in the type II Δku80 Δhxgprt strain measured at the orotate (OPRT) and the uracil (UPRT) phosphoribosyltransferase loci was highly efficient. To assess the potential of the type II Δku80 Δhxgprt strain to examine gene function affecting cyst biology and latent stages of infection, we targeted the deletion of four parasite antigen genes (GRA4, GRA6, ROP7, and tgd057) that encode characterized CD8+ T cell epitopes that elicit corresponding antigen-specific CD8+ T cell populations associated with control of infection. Cyst development in these type II mutant strains was not found to be strictly dependent on antigen-specific CD8+ T cell host responses. In contrast, a significant biological role was revealed for the dense granule proteins GRA4 and GRA6 in cyst development since brain tissue cyst burdens were drastically reduced specifically in mutant strains with GRA4 and/or GRA6 deleted. Complementation of the Δgra4 and Δgra6 mutant strains using a functional allele of the deleted GRA coding region placed under the control of the endogenous UPRT locus was found to significantly restore brain cyst burdens. These results reveal that GRA proteins play a functional role in establishing cyst burdens and latent infection. Collectively, our results suggest that a type II Δku80 Δhxgprt genetic background enables a higher-throughput functional analysis of the parasite genome to reveal fundamental aspects of parasite biology controlling virulence, pathogenesis, and transmission. PMID:21531875

  10. Formation of acellular cementum-like layers, with and without extrinsic fiber insertion, along inert bone surfaces of aging c-Src gene knockout mice.

    Science.gov (United States)

    Baba, Otto; Miyata, Atsushi; Abe, Tatsuhiko; Shibata, Shunichi; Nakano, Yukiko; Terashima, Tatsuo; Oda, Tsuyoshi; Kudo, Akira; Takano, Yoshiro

    2006-12-01

    To investigate the long-term effects of c-src deficiency on skeletal and dental tissues, we examined the lower jaws and long bones of c-src gene knockout (c-src KO) mice by histological and histochemical methods. Numerous multinucleated osteoclasts were distributed throughout the mandible in 5-wk-old c-src KO mice, but by 14 wk they had almost completely disappeared from the alveolar bone, leaving tartrate-resistant acid phosphatase (TRAP)-positive layers along the bone surface. Deposition of osteopontin-positive mineralized tissue, reminiscent of acellular afibrillar cementum (AAC), was confirmed along the TRAP-positive bone surface at 14 wk. The layer progressively thickened up to 21 months. A comparable mineralized layer was noted along the trabeculae of long bones as thickened cement lines. In the periostin-rich areas of jaw bones, but not in the long bones, portions of AAC-like mineralized layers were often replaced with and/or covered by acellular extrinsic fiber cementum (AEFC)-like tissue. These data suggest that the deposition of AAC-like mineralized tissue is a general phenomenon that may occur along inert or slowly remodeling bone surfaces under conditions characterized by reduced bone-resorbing activity, whereas the induction of AEFC-like tissue seems to be associated with the expression of certain molecules that are particularly abundant in the microenvironment of the periodontal ligament. PMID:17184236

  11. Improvements to the APEX apparatus

    Energy Technology Data Exchange (ETDEWEB)

    Ahmad, I.; Back, B.B.; Betts, R.R. [and others

    1995-08-01

    A number of technical issues led us to rework extensively the APEX apparatus in summer 1994. During the earlier runs, a significant fraction of the 432 silicon detector elements showed degraded resolution such that they had to be excluded from the final analysis in software. The effect of this is to reduce the efficiency of APEX and possibly also to introduce holes in the acceptance which, for some perhaps exotic scenarios, might reduce the acceptance to an unacceptably low level. Also, the energy thresholds below which it is not possible to generate timing information from the silicon detectors, were high enough that the low-energy acceptance of APEX was compromised to a significant extent. The origins of these difficulties were in part due to degraded performance of the silicon detectors themselves, problems with the silicon cooling systems and electronics problems. Both silicon arrays were disassembled and sub-standard detectors replaced, all detectors were also cleaned with the result that all detectors now performed at the specified values of leakage current. The silicon cooling systems were disassembled and rebuilt with the result that many small leaks were fixed. Defective electronics channels were repaired or replaced. The rotating target wheel was also improved with the installation of new bearings and a computer-controlled rotation and readout system. The rebuilt wheel can now run at speeds up to 900 rpm for weeks on end without breakdown. The target wheel and associated beam sweeping now work extremely well so that low-melting-point targets such as Pb and In can be used in quite intense beams without melting.

  12. Construction of a host range-expanded hybrid baculovirus of BmNPV and AcNPV,and knockout of cysteinase gene for more efficient expression

    Institute of Scientific and Technical Information of China (English)

    2004-01-01

    AcNPV(Autographa californica nuclear polyhedrosis virus)and BmNPV(Bombyx mori nuclear polyhedrosis virus)are two principal insect-baculovirus expression systems,each having different characteristics.AcNPV has a wider host range and can infect a series of cell lines thus making it suitable for cell suspension culture expression,but the small size of the host insect,A.californica,makes AcNPV less suitable for large scale protein synthesis.In contrast,BmNPV can only infect the silkworm,Bornbyx rnori,which is well-known for its easy rearing and large size.These characteristics make the BmNPV system especially suitable for large-scale industrial expression.To utilize the advantages of both AcNPV and BmNPV,we tried to expand their host range through homologous recombination and successfully constructed a hybrid baculovirus of AcNPV and BmNPV,designated as HyNPV.The hybrid baculovirus can infect the hosts of both AcNPV and BmNPV.Taking the human basic fibroblast growth factor(Bfgf)gene as an application example,we constructed a recombinant,HyNPV-Bfgf.This construct is able to express the Bfgf protein both in silkworm larvae and in common-use cell lines,sf21,sf9 and High-five.Moreover,to reduce the loss of recombinant protein due to degradation by proteases that are simultaneously expressed by the baculovirus,we knocked out the cysteinase gene coding for one of the most important baculovirus proteases.This knockout mutation improves the production efficiency of the Bfgf recombinant protein.

  13. Endoscopic management of orbital apex lesions.

    Science.gov (United States)

    Sethi, D S; Lau, D P

    1997-01-01

    Lesions of the orbital apex often present a diagnostic dilemma. Clinical assessment and imaging studies are helpful but a tissue biopsy is often required. The morbidity associated with transcranial approaches to the orbital apex may outweigh the benefits of obtaining a biopsy by these routes. Fine needle aspiration cytology of orbital apex lesions can be performed but there are disadvantages with this method. We describe a transnasal endoscopic technique to biopsy the orbital apex. The technique was used successfully to obtain a tissue diagnosis in six patients with orbital apex lesions. This enabled commencement of definitive treatment. There were no significant complications. The transnasal approach to the orbital apex using the endoscopes is reliable. Endoscopes provide excellent illumination, magnification, and a panoramic view of the operative field. PMID:9438058

  14. Conditional knockout of tumor overexpressed gene in mouse neurons affects RNA granule assembly, granule translation, LTP and short term habituation.

    Directory of Open Access Journals (Sweden)

    Elisa Barbarese

    Full Text Available In neurons, specific RNAs are assembled into granules, which are translated in dendrites, however the functional consequences of granule assembly are not known. Tumor overexpressed gene (TOG is a granule-associated protein containing multiple binding sites for heterogeneous nuclear ribonucleoprotein (hnRNP A2, another granule component that recognizes cis-acting sequences called hnRNP A2 response elements (A2REs present in several granule RNAs. Translation in granules is sporadic, which is believed to reflect monosomal translation, with occasional bursts, which are believed to reflect polysomal translation. In this study, TOG expression was conditionally knocked out (TOG cKO in mouse hippocampal neurons using cre/lox technology. In TOG cKO cultured neurons granule assembly and bursty translation of activity-regulated cytoskeletal associated (ARC mRNA, an A2RE RNA, are disrupted. In TOG cKO brain slices synaptic sensitivity and long term potentiation (LTP are reduced. TOG cKO mice exhibit hyperactivity, perseveration and impaired short term habituation. These results suggest that in hippocampal neurons TOG is required for granule assembly, granule translation and synaptic plasticity, and affects behavior.

  15. Oracle APEX 4.2 reporting

    CERN Document Server

    Pathak, Vishal

    2013-01-01

    Oracle APEX 4.2 Reporting is a practical tutorial for intermediate to advanced use, with plenty of step-by-step instructions and business scenarios for understanding and implementing the ins and outs of making reports.""Oracle APEX 4.2 Reporting"" is for you if you design or develop advanced solutions in APEX or wish to know about the advanced features of APEX. If you wish to have a 360 degree view of reporting technologies or work in a complex heterogeneous enterprise, this is a must-have.

  16. Observation of tail suspension test in Fmr1 gene knockout mice%Fmr1基因敲除小鼠悬尾实验的观察

    Institute of Scientific and Technical Information of China (English)

    胡丽婵; 黄海樱; 郭艺; 孙祺章; 余国汉; 黄月玲; 戴丽军; 党亚梅; 黄雄; 陈盛强

    2016-01-01

    Objective To observe tail suspension test in Fmr1 gene knockout mice and to explore whether there are differences in mobility of KO and WT mice. Methods 1 80 test mice were divided into two groups:① KO group (4,6,8 weeks old,each age group of mice is 30,male and female in half,a total of 90)② WT group (4,6,8 weeks old,each group of mice is 30,male and female on half,a total of 90).Through forced swimming test and tail suspension test to observe gender, age effect on immobility time. Results With the same age of the same sex,the KO mice’s immobility time was longer than WT mice’s.P <0.05.With the same age,the male mice’s immobility time was shorter than female mice’s.With the age in-crease,the immobility time of KO mice was longer than WT mice.P <0.05. Conclusion Fmr1 gene knockout mice have anxiety and depressive behavior.%目的:对不同周龄的 KO 小鼠与 WT 小鼠进行悬尾实验进行观察,探讨 KO 小鼠与 WT 小鼠的行为差别。方法采用健康的试验动物180只分两组:①KO 组(4、6、8周龄,各周龄30只,雌雄各半,共90只)②WT 组(4、6、8周龄,各周龄30只,雌雄各半,共90只);通过悬尾实验观察性别,年龄对不动时间的影响。结果同龄 KO 雌性小鼠比雄性小鼠的静止时间差别不大;随着年龄增大,静止时间增长。同龄同性别的 KO 鼠比 WT 鼠的不动时间长。P <0.05;同龄雄性小鼠比雌性小鼠的不动时间短;随年龄增长各种系小鼠不动时间增长,KO 鼠的不动时间比 WT 鼠长,P <0.05。结论 KO 小鼠存在抑郁行为表型。

  17. Fmr1基因敲除雄性小鼠生长指标的变化%Observation on the results of body weight and length of male mince with Fmr1 gene knockout.

    Institute of Scientific and Technical Information of China (English)

    杨乙; 刘国彬; 刘绪红; 林波; 黄月玲; 沈岩松; 张维雯; 孙卫文; 李敏雄; 陈盛强

    2011-01-01

    目的 对出生后0~56d的清洁级FVB小鼠和Fmr1基因敲除雄性小鼠的体重和体长指标进行分析比较,同时比较出生后28d的睾丸大小变化.方法 挑选10周龄FVB小鼠和Fmr1基因敲除小鼠各20只(雌、雄各半),采取1:I同居,全同胞兄妹近交繁殖,测定雄性子代生长发育指标,进行统计分析.结果 出生后0~56d清洁级Fmr1基因敲除雄性小鼠体重与体长的增长与FVB小鼠差异无统计学意义(t=0.93,t=1.24,P>0.05),但出生后28d的FVB小鼠和Fmr1基因敲除雄性小鼠睾丸大小差别有统计学意义(t=4.12,P<0.05).结论 Fmr1基因敲除不影响雄性小鼠正常的体重和体长的发育,但出生后28d的Fmr1基因敲除小鼠有巨睾征.%Objective To characterize the growth performance of FVB mice and Fmrl gene knockout mice. Methods There 20 seed FVB micedO males and 10 females,aged 10 weeks)were chosen and monogamously mated. The parameters of growth performance were analyzed. And these analysis also conducted on male Fmrl gene knockout mice. Results There no significant statistical differences in average weight and length of the newboms FVB and Fmrl gene knockout mice within 56 days after borth were observed. While significant differences were observed in size of testes of FVB mice and Fmrl gene knockout mice 28 days after birth. Conclusion Fmrl gene knockout does not affect the growth and weight of the mice but the mice may have giant testes.

  18. A rare case of petrous apex osteoma.

    Science.gov (United States)

    Cece, Hasan; Yildiz, Sema; Iynen, Ismail; Karakas, Omer; Karakas, Ekrem; Dogan, Ferit

    2012-06-01

    Osteomas are the most common tumours of the cranial vault and facial skeleton. Temporal bone osteoma is a rare entity. An osteoma arising from the petrous apex is extremely rare. We present a case of osteoma arising from the petrous apex followed by a discussion of the etiology, presentation, and radiologic findings.

  19. Petrous apex lesions in the pediatric population

    Energy Technology Data Exchange (ETDEWEB)

    Radhakrishnan, Rupa [University of Cincinnati College of Medicine, Department of Radiology, Cincinnati, OH (United States); Cincinnati Children' s Hospital Medical Center, Department of Radiology, Cincinnati, OH (United States); Son, Hwa Jung [University of Cincinnati College of Medicine, Department of Otolaryngology-Head and Neck Surgery, Cincinnati, OH (United States); Koch, Bernadette L. [Cincinnati Children' s Hospital Medical Center, Department of Radiology, Cincinnati, OH (United States)

    2014-03-15

    A variety of abnormal imaging findings of the petrous apex are encountered in children. Many petrous apex lesions are identified incidentally while images of the brain or head and neck are being obtained for indications unrelated to the temporal bone. Differential considerations of petrous apex lesions in children include ''leave me alone'' lesions, infectious or inflammatory lesions, fibro-osseous lesions, neoplasms and neoplasm-like lesions, as well as a few rare miscellaneous conditions. Some lesions are similar to those encountered in adults, and some are unique to children. Langerhans cell histiocytosis (LCH) and primary and metastatic pediatric malignancies such as neuroblastoma, rhabomyosarcoma and Ewing sarcoma are more likely to be encountered in children. Lesions such as petrous apex cholesterol granuloma, cholesteatoma and chondrosarcoma are more common in adults and are rarely a diagnostic consideration in children. We present a comprehensive pictorial review of CT and MRI appearances of pediatric petrous apex lesions. (orig.)

  20. 变形链球菌gcp基因敲除菌株表达谱基因芯片*☆%Streptococcus mutans gcp gene knockout strains expression profile gene chip

    Institute of Scientific and Technical Information of China (English)

    谢苗苗; 胡晓聪; 吴补领; 闫文娟

    2013-01-01

    BACKGROUND:Previous studies have confirmed the presence of bis-(3'-5')-cyclic dimeric guanosine monophosphate signaling pathway in Streptococcus mutans, which construct the streptococcus mutans gcp gene knockout strains. OBJECTIVE:To compare the gene expression differences between Streptococcus mutans wild strains and gcp mutant strains, and to screen the biofilm-related genes from them for the fol ow-up study. METHODS:The total RNA of two kinds of strains were extracted and stained with cy3 and cy5 respectively after reverse transcription. The gene chip was scanned after hybridization and the differential gene were obtained through the data analysis. The different expression genes were verified by real-time PCR. RESULTS AND CONCLUSION:Differential genes were mainly relative about glucose metabolism and biofilm formation. We selected two genes for real-time PCR verification. The PCR results were consistent with the microarray results. After Streptococcus mutans gcp gene knockout, the gene expressions of gcp mutant strains were upregulated and the gene expressions of phosphotransferase system were downregulated, this result suggested that two different genes were related with the c-di-GMP signal pathway downstream.%  背景:前期研究中经证实变形链球菌内部存在单磷酸鸟苷环二聚体信号通路,构建了变形链球菌gcp基因敲除菌株。  目的:比较变形链球菌野生菌种和gcp基因突变菌株基因表达的差异情况,筛选与生物膜相关的基因,进入后续研究。  方法:提取两种细菌的总RNA,反转录后分别用cy3和cy5染色。与基因芯片杂交后,扫描结果,进行数据分析,获取差异基因信息,对筛选的基因进行Real-Time PCR验证。  结果与结论:差异基因主要与糖代谢、生物膜形成有关,选择了2个基因进行验证,PCR结果与芯片结果相符合。变形链球菌gcp基因敲除后,突变菌株ahpC基因表达

  1. spd1672基因敲除显著影响肺炎链球菌的毒力%Spd1672 gene knockout significantly attenuates the virulence of Streptococcus pneumoniae

    Institute of Scientific and Technical Information of China (English)

    黄健; 黄美容; 吴凯峰; 朱杰华; 黎兵; 闵迅

    2015-01-01

    目的 研究spd1672基因在肺炎链球菌感染过程中的作用.方法 通过腹腔攻毒实验和细菌入血载量分析、细菌的粘附侵袭实验、全血杀菌实验以及相关细胞因子检测等观察spd1672基因敲除(D39△1672菌株)后对肺炎链球菌毒力的影响.结果D39△1672菌株感染组小鼠中位生存时间和生存率显著高于野生菌株组(P0.05), the spd1672 knockout strain showed significantly lower cell invasion ability than the wild-type strain (P<0.05). The spd1672 knockout strain also had a reduced resistance to whole blood cells, and thw mice infected with spd1672 knockout strain exhibit lower levels of serum inflammatory cytokines than those infected with the wild-type strain. Conclusion Spd1672 gene is importantly related to the virulence of S. pneumoniae and plays important roles in modulating bacterial invasion, resistance to whole blood cells and proinflammatory responses.

  2. Novel roles for metallothionein-I + II (MT-I + II) in defense responses, neurogenesis, and tissue restoration after traumatic brain injury: insights from global gene expression profiling in wild-type and MT-I + II knockout mice

    DEFF Research Database (Denmark)

    Penkowa, Milena; Cáceres, Mario; Borup, Rehannah;

    2006-01-01

    . A genomic approach, such as the use of microarrays, provides much insight in this regard, especially if combined with the use of gene-targeted animals. We report here the results of one of these studies comparing wild-type and metallothionein-I + II knockout mice subjected to a cryolesion...... of the somatosensorial cortex and killed at 0, 1, 4, 8, and 16 days postlesion (dpl) using Affymetrix genechips/oligonucleotide arrays interrogating approximately 10,000 different murine genes (MG_U74Av2). Hierarchical clustering analysis of these genes readily shows an orderly pattern of gene responses at specific...... and opened new avenues that were confirmed by immunohistochemistry. Data in KO, MT-I-overexpressing, and MT-II-injected mice strongly suggest a role of these proteins in postlesional activation of neural stem cells....

  3. Desenvolvimento e avaliação de uma cepa knockout de Brucella abortus obtida pela deleção do gene virB10 Development and evaluation of a strain of Brucella abortus gotten by the knockout of the virB10 gene

    Directory of Open Access Journals (Sweden)

    Fabiane G. de Souza

    2009-11-01

    Full Text Available Brucella spp. são bactérias gram-negativas, intracelulares facultativas que são patogênicas para muitas espécies de mamíferos causando a brucelose, uma zoonose difundida mundialmente. Por isso a busca de alternativas de controle mais eficientes se faz necessário como o desenvolvimento de novas cepas que possam ser testadas como potenciais imunógenos. Neste estudo realizou-se a deleção do gene virB10 da cepa S2308 de Brucella abortus gerando uma cepa knockout provavelmente incapaz de produzir a proteína nativa correspondente. O gene virB10 faz parte de um operon que codifica para um sistema de secreção do tipo IV, essencial para a sobrevivência intracelular e multiplicação da bactéria em células hospedeiras. A deleção foi realizada pela construção do plasmídeo suicida pBlue:virB10:kan e eletroporação deste em células eletrocompetentes de B. abortus S2308, ocorrendo a troca do gene selvagem pelo gene interrompido, com o gene de resistência a canamicina, por recombinação homóloga dupla. Camundongos BALB/c foram inoculados com as cepas S19, RB-51, ΔvirB10 de B. abortus e B. abortus S2308 selvagem; os resultados demonstraram que camundongos BALB/c inoculados com S19 e camundongos BALB/c inoculados com S2308 apresentaram queda mais rápida de linha de tendência, quando comparadas aos demais grupos, para recuperação bacteriana (RB e peso esplênico (PE respectivamente. Os grupos que receberam ΔvirB10 S2308 de B. abortus e RB-51 demonstraram comportamento semelhante para ambas as características. Na sexta semana após a inoculação, os resultados para RB (log de UFC ± desvio padrão e PE (peso esplênico ± desvio padrão, respectivamente, mostraram: grupos inoculados com as cepas S2308 (4,44±1,97 e 0,44±0,11, S19 (1,83±2,54 e 0,31±0,04, RB-51 (0,00±0,00 e 0,20±0,01 e ΔvirB10 S2308 (1,43±1,25 e 0,19±0,03. Considerado o clearance bacteriano, todos os grupos diferiram estatisticamente do grupo que recebeu S

  4. KnockoutJS blueprints

    CERN Document Server

    Russo, Carlo

    2015-01-01

    If you are a JavaScript developer and already know the basics of KnockoutJS and you want to get the most out of it, then this book is for you. This book will help in your transition from a small site to a large web application that is easily maintainable.

  5. [Electronic Apex Locator as a dental instrument].

    Science.gov (United States)

    Lin, S; Winocur-Arias, O; Slutzky-Goldberg, I

    2009-04-01

    Electronic Apex Locators (EAL) have become widely used in the last decade. The first apex locator was introduced in 1962, based on a constant electrical resistance (6.5 K.) between the oral mucosa and periodontal ligament. The first and second generations of EAL were inaccurate and could not detect the apex in the presence of conducting fluids. The third generation solved this problem by using two alternating frequencies and calculating the impedance between them. This provided reliable and accurate results in dry canals, or in the presence of blood, electrolytes or other fluid in the root canals, when the pulp was necrotic or when there was a perforation along the root. The Root ZX and Apit (Endex) are the most documented devices. The new fourth generation of apex locators is a diverse group: some use multifrequency currents, others use a "lookup matrix" rather than calculate the readings. Several of the newer EALs are smaller, and others connect to computers. PMID:20162984

  6. Asymmetric pneumatization of the petrous apex.

    Science.gov (United States)

    Roland, P S; Meyerhoff, W L; Judge, L O; Mickey, B E

    1990-07-01

    Three patients with high-intensity MR signals from one petrous apex, but nonpathologic fine-cut computed tomography are reported. In two of the three patients, normal bone marrow within the petrous apex on one side is believed to have generated the high-intensity signal. In one of the three patients, the etiology of the MR image remains obscure, but may represent the earliest stages of petrous cholesterol granuloma or mucocele. We have reviewed 500 head CT scans performed for non-otologic reasons, in an attempt to establish the frequency of this finding. The literature on MR and CT imaging of the petrous apex and asymmetric pneumatization of the petrous apex is reviewed. PMID:2117735

  7. Apex Predators Program Age and Growth Data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Apex Predators Program staff have collected vertebral centra from sportfishing tournaments, cruises, commercial fishermen and strandings in the Northeast US since...

  8. Apex Predators Program Sportfishing Tournament Data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Apex Predators Program staff have collected shark sportfishing tournamant data from the Northeast US since the 1960's. These tournaments offer a unique opportunity...

  9. Leukemogenesis in heterozygous PU.1 knockout mice.

    Science.gov (United States)

    Genik, Paula C; Vyazunova, Irina; Steffen, Leta S; Bacher, Jeffery W; Bielefeldt-Ohmann, Helle; McKercher, Scott; Ullrich, Robert L; Fallgren, Christina M; Weil, Michael M; Ray, F Andrew

    2014-09-01

    Most murine radiation-induced acute myeloid leukemias involve biallelic inactivation of the PU.1 gene, with one allele being lost through a radiation-induced chromosomal deletion and the other allele affected by a recurrent point mutation in codon 235 that is likely to be spontaneous. The short latencies of acute myeloid leukemias occurring in nonirradiated mice engineered with PU.1 conditional knockout or knockdown alleles suggest that once both copies of PU.1 have been lost any other steps involved in leukemogenesis occur rapidly. Yet, spontaneous acute myeloid leukemias have not been reported in mice heterozygous for a PU.1 knockout allele, an observation that conflicts with the understanding that the PU.1 codon 235 mutation is spontaneous. Here we describe experiments that show that the lack of spontaneous leukemia in PU.1 heterozygous knockout mice is not due to insufficient monitoring times or mouse numbers or the genetic background of the knockout mice. The results reveal that spontaneous leukemias that develop in mice of the mixed 129S2/SvPas and C57BL/6 background of knockout mice arise by a pathway that does not involve biallelic PU.1 mutation. In addition, the latency of radiation-induced leukemia in PU.1 heterozygous mice on a genetic background susceptible to radiation-induced leukemia indicates that the codon 235 mutation is not a rate-limiting step in radiation leukemogenesis driven by PU.1 loss.

  10. The Large APEX Bolometer Camera LABOCA

    CERN Document Server

    Siringo, G; Kovács, A; Schuller, F; Weiss, A; Esch, W; Gemuend, H P; Jethava, N; Lundershausen, G; Colin, A; Guesten, R; Menten, K M; Beelen, A; Bertoldi, F; Beeman, J W; Haller, E E

    2009-01-01

    The Large APEX Bolometer Camera, LABOCA, has been commissioned for operation as a new facility instrument t the Atacama Pathfinder Experiment 12m submillimeter telescope. This new 295-bolometer total power camera, operating in the 870 micron atmospheric window, combined with the high efficiency of APEX and the excellent atmospheric transmission at the site, offers unprecedented capability in mapping submillimeter continuum emission for a wide range of astronomical purposes.

  11. Petrous apex mucocele: high resolution CT.

    Science.gov (United States)

    Memis, A; Memis, A; Alper, H; Calli, C; Ozer, H; Ozdamar, N

    1994-11-01

    Mucocele of the petrous apex is very rare, only three cases having been reported. Since this area is inaccessible to direct examination, imaging, preferably high resolution computed tomography (HR CT) is essential. We report a case showing an eroding, non enhancing mass with sharp, lobulated contours, within the petrous apex. The presence of a large air cell on the opposite side suggested a mucocele. PMID:7862284

  12. Petrous apex mucocele: high resolution CT

    Energy Technology Data Exchange (ETDEWEB)

    Memis, A. [Dept. of Radiology, Hospital of Ege Univ., Bornova, Izmir (Turkey); Memis, A. [Dept. of Radiology, Hospital of Ege Univ., Bornova, Izmir (Turkey); Alper, H. [Dept. of Radiology, Hospital of Ege Univ., Bornova, Izmir (Turkey); Calli, C. [Dept. of Radiology, Hospital of Ege Univ., Bornova, Izmir (Turkey); Ozer, H. [Dept. of Radiology, Hospital of Ege Univ., Bornova, Izmir (Turkey); Ozdamar, N. [Dept. of Neurosurgery, Hospital of Ege Univ., Bornova, Izmir (Turkey)

    1994-11-01

    Mucocele of the petrous apex is very rare, only three cases having been reported. Since this area is inaccessible to direct examination, imaging, preferably high resolution computed tomography (HR CT) is essential. We report a case showing an eroding, non enhancing mass with sharp, lobulated contours, within the petrous apex. The presence of a large air cell on the opposite side suggested a mucocele. (orig.)

  13. Petrous apex mucocele: high resolution CT

    International Nuclear Information System (INIS)

    Mucocele of the petrous apex is very rare, only three cases having been reported. Since this area is inaccessible to direct examination, imaging, preferably high resolution computed tomography (HR CT) is essential. We report a case showing an eroding, non enhancing mass with sharp, lobulated contours, within the petrous apex. The presence of a large air cell on the opposite side suggested a mucocele. (orig.)

  14. Hormone-Sensitive Lipase Knockouts

    Directory of Open Access Journals (Sweden)

    Shen Wen-Jun

    2006-02-01

    Full Text Available Abstract All treatments for obesity, including dietary restriction of carbohydrates, have a goal of reducing the storage of fat in adipocytes. The chief enzyme responsible for the mobilization of FFA from adipose tissue, i.e., lipolysis, is thought to be hormone-sensitive lipase (HSL. Studies of HSL knockouts have provided important insights into the functional significance of HSL and into adipose metabolism in general. Studies have provided evidence that HSL, though possessing triacylglycerol lipase activity, appears to be the rate-limiting enzyme for cholesteryl ester and diacylglycerol hydrolysis in adipose tissue and is essential for complete hormone stimulated lipolysis, but other triacylglycerol lipases are important in mediating triacylglycerol hydrolysis in lipolysis. HSL knockouts are resistant to both high fat diet-induced and genetic obesity, displaying reduced quantities of white with increased amounts of brown adipose tissue, increased numbers of adipose macrophages, and have multiple alterations in the expression of genes involved in adipose differentiation, including transcription factors, markers of adipocyte differentiation, and enzymes of fatty acid and triglyceride synthesis. With disruption of lipolysis by removal of HSL, there is a drastic reduction in lipogenesis and alteration in adipose metabolism.

  15. Construction of Deletion-knockout Mutant Fowlpox Virus (FWPV)

    OpenAIRE

    Laidlaw, Stephen M.; Skinner, Michael A.

    2014-01-01

    The construction of deletion-knockout poxviruses is a useful approach to determining the function of specific virus genes. This protocol is an adaptation of the transient dominant knockout selection protocol published by Falkner and Moss (1990) for use with vaccinia virus. The protocol makes use of the dominant selectable marker Escherichia coli guanine phosphoribosyltransferase (gpt) gene (Mulligan and Berg, 1981), under the control of an early/late poxvirus promoter. The deletion viruses th...

  16. Researching and deploying an APEX security scanning tool

    CERN Document Server

    Vali, Silvia

    2016-01-01

    Most of the APEX applications have not been developed considering security in mind or were developed many years ago, as well as the old version of APEX used exposes those type of applications to a variety of potential security risks. CERN develops and uses many APEX applications, but none of the currently used tools provides a sufficient way of vulnerability scanning for such applications. The current version of APEX used in CERN is 4.2.6 whilst the latest version is 5.1. This report provides the reader with the overview on APEX and the APEX-SERT vulnerability scanning tool as well as the summary of testing the APEX-SERT tool on existing APEX applications used in CERN and the samples, created during this project. The goal of this project was to research on existing tools for vulnerability scanning of APEX applications and to deploy the tool to be used APEX developers.

  17. KnockoutJS essentials

    CERN Document Server

    Ferrando, Jorge

    2015-01-01

    If you are a JavaScript developer who has been using DOM manipulation libraries such as Mootools or Scriptaculous, and you want go further in modern JavaScript development with a simple and well-documented library, then this book is for you. Learning how to use Knockout will be perfect as your next step towards building JavaScript applications that respond to user interaction.

  18. 大中型动物基因敲除技术的研究进展%The Development of Gene Knockout Technologies in Large and Medium Animal Models

    Institute of Scientific and Technical Information of China (English)

    刘雪静; 王欢; 严放; 高明明; 刘国庆; 黄薇

    2015-01-01

    基因敲除是20世纪80年代末发展起来的一门新技术,2007年获得诺贝尔生理或医学奖。然而在很长一段时间内,由于胚胎干细胞(embryonic stem cell,ES cell)体外的成功培养仅限于小鼠,该技术很难在其它动物种系中完成。2008年至今随着新 ES 细胞基因打靶、锌指核酸酶(zinc finger nucleases,ZFNs)、转录激活因子样效应物核酸酶(transcription activator-like effect nucleases, TALENs)和规律成簇的间隔短回文重复序列/Cas9内切酶(clusters of regularly interspased short pal-indromic repeats/Cas9,CRISPR/Cas9)等新技术的不断涌现,使在疾病研究中更接近于人类的大中型动物基因敲除成为可能。本综述将介绍近几年来以上几种基因敲除新技术在大中型动物上的成功应用及重大意义。%Technique of homologous recombination based gene targeting developed in the late 1 980s and won the Nobel Prize in Physiology or Medicine in 2007.However,this technique could only performed in mice which embryonic stem (ES)cells could keep in the potential of multifunction in vitro.Therefore gene knockout technology was difficult to be applied in other species of animals for a long time.Since 2008,with the development of the new technologies,such as the new ES cell gene targeting,zinc finger nucleases (ZFNs),transcription activator-like effector nucleases (TALENs)and clusters of regularly in-terspaced short palindromic repeats/Cas9 (CRISPR/Cas9),building gene knockout in large and medium animals models which are similar to human in disease research becomes possible.This review describes some new gene knockout technologies in large and medium animal models for recent years.

  19. Ultra-superovulation for the CRISPR-Cas9-mediated production of gene-knockout, single-amino-acid-substituted, and floxed mice.

    Science.gov (United States)

    Nakagawa, Yoshiko; Sakuma, Tetsushi; Nishimichi, Norihisa; Yokosaki, Yasuyuki; Yanaka, Noriyuki; Takeo, Toru; Nakagata, Naomi; Yamamoto, Takashi

    2016-08-15

    Current advances in producing genetically modified mice using genome-editing technologies have indicated the need for improvement of limiting factors including zygote collection for microinjection and their cryopreservation. Recently, we developed a novel superovulation technique using inhibin antiserum and equine chorionic gonadotropin to promote follicle growth. This method enabled the increased production of fertilized oocytes via in vitro fertilization compared with the conventional superovulation method. Here, we verify that the ultra-superovulation technique can be used for the efficient generation of clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 9 (Cas9)-mediated knockout mice by microinjection of plasmid vector or ribonucleoprotein into zygotes. We also investigated whether single-amino-acid-substituted mice and conditional knockout mice could be generated. Founder mice bearing base substitutions were generated more efficiently by co-microinjection of Cas9 protein, a guide RNA and single-stranded oligodeoxynucleotide (ssODN) than by plasmid microinjection with ssODN. The conditional allele was successfully introduced by the one-step insertion of an ssODN designed to carry an exon flanked by two loxP sequences and homology arms using a double-cut CRISPR-Cas9 strategy. Our study presents a useful method for the CRISPR-Cas9-based generation of genetically modified mice from the viewpoints of animal welfare and work efficiency.

  20. Comparison of brown and white adipose tissue fat fractions in ob, seipin, and Fsp27 gene knockout mice by chemical shift-selective imaging and (1)H-MR spectroscopy.

    Science.gov (United States)

    Peng, Xin-Gui; Ju, Shenghong; Fang, Fang; Wang, Yu; Fang, Ke; Cui, Xin; Liu, George; Li, Peng; Mao, Hui; Teng, Gao-Jun

    2013-01-15

    Brown adipose tissue (BAT) plays a key role in thermogenesis to protect the body from cold and obesity. White adipose tissue (WAT) stores excess energy in the form of triglycerides. To better understand the genetic effect on regulation of WAT and BAT, we investigated the fat fraction (FF) in two types of adipose tissues in ob/ob, human BSCL2/seipin gene knockout (SKO), Fsp27 gene knockout (Fsp27(-/-)), and wild-type (WT) mice in vivo using chemical shift selective imaging and (1)H-MR spectroscopy. We reported that the visceral fat volume in WAT was significantly larger in ob/ob mice, but visceral fat volumes were lower in SKO and Fsp27(-/-) mice compared with WT mice. BAT FF was significantly higher in ob/ob mice than the WT group and similar to that of WAT. In contrast, WAT FFs in SKO and Fsp27(-/-) mice were lower and similar to that of BAT. The adipocyte size of WAT in ob/ob mice and the BAT adipocyte size in ob/ob, SKO, and Fsp27 mice were significantly larger compared with WT mice. However, the WAT adipocyte size was significantly smaller in SKO mice than in WT mice. Positive correlations were observed between the adipocyte size and FFs of WAT and BAT. These results suggested that smaller adipocyte size correlates with lower FFs of WAT and BAT. In addition, the differences in FFs in WAT and BAT measured by MR methods in different mouse models were related to the different regulation effects of ob, seipin, or Fsp27 gene on developing WAT and BAT.

  1. 补体C3基因敲除小鼠的繁育及子代基因型鉴定%Reproduction and Genotype Identification of Complement 3 Gene Knockout Mice

    Institute of Scientific and Technical Information of China (English)

    郑静; 刘阳; 胡松; 李敏惠; 李丽梅; 阳泰; 杨淑霞; 邹强

    2011-01-01

    Objective This study was to explore the method to breed and identify complement 3 gene knockout mice. Methods The heterozygote of complement 3 gene knockout mice ( C3 + / - ) were bred and reproduced.Wild genotype ( C3 + / + ), heterozygote genotype ( C3 + / - ) , and homozygote genotype ( C3 - / - ) would appear in offspring. The genotypes were characterized using ELISA and PCR. Result The heterozygote mice bred 135 offspring. 38 mice are wild type ,68 mice are heterozygote, and 29 mice are homozygote. By x2text, the segregate ratio of offspring was fitted to the Mendelian ratio. Conclusion Appropriate methods for breeding, reproducing, and identifying are the effective way for acquiring complement 3 gene knockout mice from heterozygote.%目的 探讨繁殖和鉴定补体C3基因敲除小鼠的实验方法.方法 将所引进的补体C3基因敲除杂合子小鼠(C3+/-)进行饲养并繁殖,其子代出现三种基因型的小鼠,即纯合子C3-/-、杂合子C3+/-、野生型C3+/+,采用ELISA与PCR相结合对子代小鼠基因型进行鉴定.结果 繁育出135只子代小鼠,经鉴定,C3+/+、C3+/-、C3-/-各为38只、68只、29只,经x2检验,子代小鼠分离比例符合孟德尔遗传规律.结论 正确的饲养繁殖以及子代鉴定是从杂合子小鼠中获得补体C3基因敲除小鼠的有效途径.

  2. Cluster knockout reactions

    Indian Academy of Sciences (India)

    Arun K Jain; B N Joshi

    2014-04-01

    Cluster knockout reactions are expected to reveal the amount of clustering (such as that of , d and even of heavier clusters such as 12C, 16O etc.) in the target nucleus. In simple terms, incident medium high-energy nuclear projectile interacts strongly with the cluster (present in the target nucleus) as if it were existing as a free entity. Theoretically, the relatively softer interactions of the two outgoing particles with the residual nucleus lead to optical distortions and are treated in terms of distorted wave (DW) formalism. The long-range projectile–cluster interaction is accounted for, in terms of the finite range (FR) direct reaction formalism, as against the more commonly adopted zero-range (ZR) distorted wave impulse approximation (DWIA) formalism. Comparison of the DWIA calculations with the observed data provide information about the momentum distribution and the clustering spectroscopic factor of the target nucleus. Interesting results and some recent advancements in the area of (, 2) reactions and heavy cluster knockout reactions are discussed. Importance of the finite-range vertex and the final-state interactions are brought out.

  3. Petrous apex lesions outcome in 21 cases

    Directory of Open Access Journals (Sweden)

    Hekmatara M

    1997-09-01

    Full Text Available Petrous apex lesions of temporal bone progress slowly. Most of the time not only destruct this area but also involve neighbouring element. The symptoms of the neighbouring neuro-vasculare involvement we can recognize these lesions. The most common symptoms of involvement of the petrous apex are: headache, conductive hearing loss or sensorineural type, paresthesia and anesthesia of the trigeminal nerve, paresia and paralysis of the facial nerve, abducent nerve. In retrospective study which has been in the ENT and HNS wards of Amiralam hospital, 148 patients have been operated due to temporal bone tumor; from these numbers, 21 (13.6% patients had petrous apex lesions of temporal bone. Eleven (52.9% patients of these 21 persons were men and the remaining 10 (47-6% were women. The average age of the patients was 37 years. The common pathology of these patients were glomus jugulare tumors, hemangioma, schwannoma, meningioma, congenital cholesteatoma, giant cell granuloma. The kind of operations that have been done on these patients were: infratemporal, translabyrinthine and middle fossa approaches. The conclusion of this study shows that petrous apex area is an occult site. The symptoms of this lesion are not characteristic, meticulous attention to the history and physical examination are very helpful to recognition of these lesions and it's extention.

  4. [Giant cholesterol cysts of the petrous apex].

    Science.gov (United States)

    Pellet, W; Valenzuela, S; Malca, S; Cannoni, M; Perez-Castillo, A M

    1992-01-01

    In connection with their two own cases, the authors deal about the giant cholesterol cysts of the petrous apex. The lesions which are to be differentiated from epidermoid cysts are cholesterol granulomas. Their petrous apex location explains their characteristic large appearance. As each cholesterol granuloma, they occur when a bony cell is obstructed. This chronic obstruction induces mucosal edema then bleedings which lead to the formation and, by the lack of drainage, to the accumulation of cholesterol crystals. These crystals initiate a non specific reaction to foreign bodies, a granuloma, which also can bleed. Thus, a continuous cycle perpetuates the growth of the lesion. This lesion, when it is localized in the petrous apex, can reach a big size before the appearance of some signs. Usually, these are otologic (sensorineural hearing loss, tinnitus, vertigo) and/or cranial nerve palsies (V, VI, VII). C.T. scan (well defined, sharply marginated bony expansible lesion with isodense to the brain central part) and M.R.I. (central region of increased intensity on both T1 and T2 weighted images and peripheral rim of markedly decreased signal intensity in all instances) features are characteristic enough to allow diagnose with other petrous apex lesions (cholesteatoma, mucocele, epithelial cyst, histiocytosis X, ...). Surgical treatment must try to evacuate and to aerate the cavity or perhaps to obliterate it with fatty pieces in order to prevent the recurrence. PMID:1299772

  5. Manipulation of Mouse Embryonic Stem Cells for Knockout Mouse Production

    OpenAIRE

    Limaye, Advait; Hall, Bradford; Kulkarni, Ashok B.

    2009-01-01

    The establishment of mouse embryonic stem (ES) cell liness has allowed for the generation of the knockout mouse. ES cells that are genetically altered in culture can then be manipulated to derive a whole mouse containing the desired mutation. To successfully generate a knockout mouse, however, the ES cells must be carefully cultivated in a pluripotent state throughout the gene targeting experiment. This unit describes detailed step-by-step protocols, reagents, equipment, and strategies needed...

  6. Smad3基因剔除小鼠的基因型鉴定与繁殖性能研究%Research of the Genotype Identification and Riproductive Performance of Smad3 Gene Knockout Mice

    Institute of Scientific and Technical Information of China (English)

    孙岩松; 吕雅歆; 王冬平; 方厚华; 时彦胜; 战大伟; 张爱兰; 李桂军

    2003-01-01

    The previous study of Srnad3 gene knockout mice ( Smad3ex8/ex8 ) shows that the Smad3ex8/ex8 mice develop progressive leukocytosis, periodontitis, gastritis, colitis and chronic infection with abscess formation adjacent to mucosal surfaces . symptomatic mutant mice exhibit thymic involution, enlarged lymph nodes and T cells with activated phenotype.Further study suggests that the thymic cells and peripheral T ceels of Smad3ex8/ex8 mice have lost the response to TGF-β.Furthermore, nwe found that the homologous Smad3ex8/ex mice developed degenerative joint disease resembling human osteoarthritis, osteoporosis and wound healing up quicker. So the mice can serve as an ideal animal model for immune dysregulation, osteoarthritis and so on.

  7. Hearing Outcomes after Surgical Drainage of Petrous Apex Cholesterol Granuloma

    OpenAIRE

    Rihani, Jordan; Kutz, J. Walter; Isaacson, Brandon

    2014-01-01

    Objective This study aims to assess the hearing outcomes of patients undergoing surgical management of petrous apex cholesterol granuloma and to discuss the role of otic capsule–sparing approaches in drainage of petrous apex cholesterol granulomas.

  8. Effect of dietary calcium and 1,25-(OH)2D3 on the expression of calcium transport genes in calbindin-D9k and -D28k double knockout mice.

    Science.gov (United States)

    Ko, Sang-Hwan; Choi, Kyung-Chul; Oh, Goo Taeg; Jeung, Eui-Bae

    2009-02-01

    The phenotypes of calbindin-D9k (CaBP-9k) and -28k (CaBP-28k) single knockout (KO) mice are similar to wild-type (WT) mice due to the compensatory action of other calcium transport proteins. In this study, we generated CaBP-9k/CaBP-28k double knockout (DKO) mice in order to investigate the importance of CaBP-9k and CaBP-28k in active calcium processing. Under normal dietary conditions, DKO mice did not exhibit any changes in phenotype or the expression of active calcium transport genes as compared to WT or CaBP-28k KO mice. Under calcium-deficient dietary conditions, the phenotype and expression of calcium transport genes in CaBP-28k KO mice were similar to WT, whereas in DKO mice, serum calcium levels and bone length were decreased. The intestinal and renal expression of transient receptor potential vanilloid member 6 (TRPV6) mRNA was significantly decreased in DKO mice fed a calcium-deficient diet as compared to CaBP-28k KO or WT mice, and DKO mice died after 4 weeks on a calcium-deficient diet. Body weight, bone mineral density (BMD) and bone length were significantly reduced in all mice fed a calcium and 1,25-(OH)(2)D(3)-deficient diet, as compared to a normal diet, and none of the mice survived more than 4 weeks. These results indicate that deletion of CaBP-28k alone does not affect body calcium homeostasis, but that deletion of CaBP-9k and CaBP-28k has a significant effect on calcium processing under calcium-deficient conditions, confirming the importance of dietary calcium and 1,25-(OH)(2)D(3) during growth and development.

  9. Rhabdomyolysis Presenting as Orbital Apex Syndrome.

    Science.gov (United States)

    Wi, Jae Min; Chi, Mijung

    2016-01-01

    Rhabdomyolysis is a condition in which striated muscle tissue breaks down rapidly and releases muscular cell constituents into extracellular fluid and the circulation. Renal symptoms, such as acute renal failure, are major complications of rhabdomyolysis. However, no previous report of rhabdomyolysis associated with orbital complication has been issued. Here, we report the first patient of rhabdomyolysis presenting as orbital apex syndrome. A 66-year-old man presented with right periorbital swelling with erythematous patches and conjunctival chemosis. In addition, swelling, redness, and vesicles were observed in both lower legs. He was found in a drunken state with the right side of his face pressed against a table. Ophthalmic examination showed right eye fixation in all directions and ischemic change of retina. Blood testing showed elevated muscle enzyme associated with muscle destruction. And computed tomography of the orbit showed swelling of right extraocular muscles and crowding of right orbital apex. Under a diagnosis of rhabdomyolysis-associated orbital apex syndrome and central retinal artery occlusion, intravenous steroid and antibiotics therapy with intraocular pressure-lowering topicals were begun. Clinical presentation, treatment course, and follow-up are discussed.

  10. Cathepsin L Plays a Major Role in Cholecystokinin Production in Mouse Brain Cortex and in Pituitary AtT-20 Cells: Protease Gene Knockout and Inhibitor Studies

    Science.gov (United States)

    Beinfeld, Margery C.; Funkelstein, Lydiane; Foulon, Thierry; Cadel, Sandrine; Kitagawa, Kouki; Toneff, Thomas; Reinheckel, Thomas; Peters, Christoph; Hook, Vivian

    2009-01-01

    Cholecystokinin (CCK) is a peptide neurotransmitter whose production requires proteolytic processing of the proCCK precursor to generate active CCK8 neuropeptide in brain. This study demonstrates the significant role of the cysteine protease cathepsin L for CCK8 production. In cathepsin L knockout (KO) mice, CCK8 levels were substantially reduced in brain cortex by an average of 75%. To evaluate the role of cathepsin L in producing CCK in the regulated secretory pathway of neuroendocrine cells, pituitary AtT-20 cells that stably produce CCK were treated with the specific cathepsin L inhibitor, CLIK-148. CLIK-148 inhibitor treatment resulted in decreased amounts of CCK secreted from the regulated secretory pathway of AtT-20 cells. CLIK-148 also reduced cellular levels of CCK9 (Arg-CCK8), consistent with CCK9 as an intermediate product of cathepsin L, shown by the decreased ratio of CCK9/CCK8. The decreased CCK0/CCK8 ratio also suggests a shift in the production to CCK8 over CCK9 during inhibition of cathepsin L. During reduction of the PC1/3 processing enzyme by siRNA, the ratio of CCK9/CCK8 was increased, suggesting a shift to the cathepsin L pathway for production of CCK9. The changes in ratios of CCK9 compared to CCK8 are consistent with dual roles of the cathepsin L protease pathway that includes aminopeptidase B to remove NH2-terminal Arg or Lys, and the PC1/3 protease pathway. These results suggest that cathepsin L functions as a major protease responsible for CCK8 production in mouse brain cortex, and participates with PC1/3 for CCK8 production in pituitary cells. PMID:19589362

  11. Understanding the progression of atherosclerosis through gene profiling and co-expression network analysis in Apob(tm2Sgy)Ldlr(tm1Her) double knockout mice.

    Science.gov (United States)

    Deshpande, Vrushali; Sharma, Ankit; Mukhopadhyay, Rupak; Thota, Lakshmi Narasimha Rao; Ghatge, Madankumar; Vangala, Rajani Kanth; Kakkar, Vijay V; Mundkur, Lakshmi

    2016-06-01

    The objective of the study was to gain molecular insights into the progression of atherosclerosis in Apob(tm2Sgy)Ldlr(tm1Her) mice, using transcriptome profiles. Weighted gene co network analysis (WGCNA) and time course analysis using limma were used to study disease progression from 0 to 20weeks. Five co-expression modules were identified by WGCNA using the expression values of 2153 genes. Genes associated with autophagy, endoplasmic reticulum stress, inflammation and lipid metabolism were differentially expressed at early stages of atherosclerosis. Time course analysis highlighted activation of inflammatory gene signaling at 4weeks, cell proliferation and calcification at 8weeks, amyloid like structures and oxidative stress at 14weeks and enhanced production of inflammatory cytokines at 20weeks. Our results suggest that maximum gene perturbations occur during early atherosclerosis which could be the danger signals associated with subclinical disease. Understanding these genes and associated pathways can help in improvement of diagnostic and therapeutic targets for atherosclerosis. PMID:27133569

  12. NR4A1 (Nur77 mediates thyrotropin-releasing hormone-induced stimulation of transcription of the thyrotropin β gene: analysis of TRH knockout mice.

    Directory of Open Access Journals (Sweden)

    Yasuyo Nakajima

    Full Text Available Thyrotropin-releasing hormone (TRH is a major stimulator of thyrotropin-stimulating hormone (TSH synthesis in the anterior pituitary, though precisely how TRH stimulates the TSHβ gene remains unclear. Analysis of TRH-deficient mice differing in thyroid hormone status demonstrated that TRH was critical for the basal activity and responsiveness to thyroid hormone of the TSHβ gene. cDNA microarray and K-means cluster analyses with pituitaries from wild-type mice, TRH-deficient mice and TRH-deficient mice with thyroid hormone replacement revealed that the largest and most consistent decrease in expression in the absence of TRH and on supplementation with thyroid hormone was shown by the TSHβ gene, and the NR4A1 gene belonged to the same cluster as and showed a similar expression profile to the TSHβ gene. Immunohistochemical analysis demonstrated that NR4A1 was expressed not only in ACTH- and FSH- producing cells but also in thyrotrophs and the expression was remarkably reduced in TRH-deficient pituitary. Furthermore, experiments in vitro demonstrated that incubation with TRH in GH4C1 cells increased the endogenous NR4A1 mRNA level by approximately 50-fold within one hour, and this stimulation was inhibited by inhibitors for PKC and ERK1/2. Western blot analysis confirmed that TRH increased NR4A1 expression within 2 h. A series of deletions of the promoter demonstrated that the region between bp -138 and +37 of the TSHβ gene was responsible for the TRH-induced stimulation, and Chip analysis revealed that NR4A1 was recruited to this region. Conversely, knockdown of NR4A1 by siRNA led to a significant reduction in TRH-induced TSHβ promoter activity. Furthermore, TRH stimulated NR4A1 promoter activity through the TRH receptor. These findings demonstrated that 1 TRH is a highly specific regulator of the TSHβ gene, and 2 TRH mediated induction of the TSHβ gene, at least in part by sequential stimulation of the NR4A1-TSHβ genes through a PKC and

  13. Regulation of gene expression by dietary Ca2+ in kidneys of 25-hydroxyvitamin D3-1 alpha-hydroxylase knockout mice.

    NARCIS (Netherlands)

    Hoenderop, J.G.J.; Chon, H.; Gkika, D.; Bluyssen, H.A.; Holstege, F.C.; St. Arnaud, R.; Braam, B.; Bindels, R.J.M.

    2004-01-01

    BACKGROUND: Pseudovitamin D deficiency rickets (PDDR) is an autosomal disease, characterized by undetectable levels of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3), rickets and secondary hyperparathyroidism. Mice in which the 25-hydroxyvitamin D3-1 alpha-hydroxylase (1 alpha-OHase) gene was inactivated, p

  14. Dietary calcium and 1,25-dihydroxyvitamin D3 regulate transcription of calcium transporter genes in calbindin-D9k knockout mice.

    Science.gov (United States)

    Ko, Sang-Hwan; Lee, Geun-Shik; Vo, Thuy T B; Jung, Eui-Man; Choi, Kyung-Chul; Cheung, Ki-Wha; Kim, Jae Wha; Park, Jong-Gil; Oh, Goo Taeg; Jeung, Eui-Bae

    2009-04-01

    The effect(s) of oral calcium and vitamin D(3) were examined on the expression of duodenal and renal active calcium transport genes, i.e., calbindin-D9k (CaBP-9k) and calbindin-D28k (CaBP-28k), transient receptor potential cation channels (TRPV5 and TRPV6), Na(+)/Ca(2+) exchanger 1 (NCX1) and plasma membrane calcium ATPase 1b (PMCA1b), in CaBP-9k KO mice. Wild-type (WT) and KO mice were provided with calcium and vitamin D(3)-deficient diets for 10 weeks. The deficient diet significantly decreased body weights compared with the normal diet groups. The serum calcium concentration of the WT mice was decreased by the deficient diet but was unchanged in the KO mice. The deficient diet significantly increased duodenal transcription of CaBP-9k and TRPV6 in the WT mice, but no alteration was observed in the KO mice. In the kidney, the deficient diet significantly increased renal transcripts of CaBP-9k, TRPV6, PMCA1b, CaBP-28k and TRPV5 in the WT mice but did not alter calcium-relating genes in the KO mice. Two potential mediators of calcium-processing genes, vitamin D receptor (VDR) and parathyroid hormone receptor (PTHR), have been suggested to be useful for elucidating these differential regulations in the calcium-related genes of the KO mice. Expression of VDR was not significantly affected by diet or the KO mutation. Renal PTHR mRNA levels were reduced by the diet, and reduced expression was also seen in the KO mice given the normal diet. Taken together, these results suggest that the active calcium transporting genes in KO mice may have resistance to the deficiency diet of calcium and vitamin D(3).

  15. Knockout of the dhfr-ts gene in Trypanosoma cruzi generates attenuated parasites able to confer protection against a virulent challenge.

    Directory of Open Access Journals (Sweden)

    Cecilia Perez Brandan

    2011-12-01

    Full Text Available BACKGROUND: Trypanosoma cruzi is a protozoan parasite that causes severe disease in millions of habitants of developing countries. Currently there is no vaccine to prevent this disease and the available drugs have the consequences of side effects. Live vaccines are likely to be more effective in inducing protection than recombinant proteins or DNA vaccines; however, safety problems associated to their use have been pointed out. In recent years, increasing knowledge on the molecular genetics of Trypanosomes has allowed the identification and elimination of genes that may be necessary for parasite infectivity and survival. In this sense, targeted deletion or disruption of specific genes in the parasite genome may protect against such reversion to virulent genotypes. METHODS AND FINDINGS: By targeted gene disruption we generated monoallelic mutant parasites for the dhfr-ts gene in a T. cruzi strain that has been shown to be naturally attenuated. In comparison to T. cruzi wild type epimastigotes, impairment in growth of dhfr-ts(+/- mutant parasites was observed and mutant clones displayed decreased virulence in mice. Also, a lower number of T. cruzi-specific CD8(+ T cells, in comparison to those induced by wild type parasites, was detected in mice infected with mutant parasites. However, no remarkable differences in the protective effect of TCC wild type versus TCC mutant parasites were observed. Mice challenged with virulent parasites a year after the original infection with the mutant parasites still displayed a significant control over the secondary infection. CONCLUSION: This study indicates that it is possible to generate genetically attenuated T. cruzi parasites able to confer protection against further T. cruzi infections.

  16. CRISPR/Cas9 mediated knockout of the abdominal-A homeotic gene in the global pest, diamondback moth (Plutella xylostella).

    Science.gov (United States)

    Huang, Yuping; Chen, Yazhou; Zeng, Baosheng; Wang, Yajun; James, Anthony A; Gurr, Geoff M; Yang, Guang; Lin, Xijian; Huang, Yongping; You, Minsheng

    2016-08-01

    The diamondback moth, Plutella xylostella (L.), is a worldwide agricultural pest that has developed resistance to multiple classes of insecticides. Genetics-based approaches show promise as alternative pest management approaches but require functional studies to identify suitable gene targets. Here we use the CRISPR/Cas9 system to target a gene, abdominal-A, which has an important role in determining the identity and functionality of abdominal segments. We report that P. xylostella abdominal-A (Pxabd-A) has two structurally-similar splice isoforms (A and B) that differ only in the length of exon II, with 15 additional nucleotides in isoform A. Pxabd-A transcripts were detected in all developmental stages, and particularly in pupae and adults. CRISPR/Cas9-based mutagenesis of Pxabd-A exon I produced 91% chimeric mutants following injection of 448 eggs. Phenotypes with abnormal prolegs and malformed segments were visible in hatched larvae and unhatched embryos, and various defects were inherited by the next generation (G1). Genotyping of mutants demonstrated several mutations at the Pxabd-A genomic locus. The results indicate that a series of insertions and deletions were induced in the Pxabd-A locus, not only in G0 survivors but also in G1 individuals, and this provides a foundation for genome editing. Our study demonstrates the utility of the CRISPR/Cas9 system for targeting genes in an agricultural pest and therefore provides a foundation the development of novel pest management tools. PMID:27318252

  17. Geometric correction of APEX hyperspectral data

    Directory of Open Access Journals (Sweden)

    Vreys Kristin

    2016-03-01

    Full Text Available Hyperspectral imagery originating from airborne sensors is nowadays widely used for the detailed characterization of land surface. The correct mapping of the pixel positions to ground locations largely contributes to the success of the applications. Accurate geometric correction, also referred to as “orthorectification”, is thus an important prerequisite which must be performed prior to using airborne imagery for evaluations like change detection, or mapping or overlaying the imagery with existing data sets or maps. A so-called “ortho-image” provides an accurate representation of the earth’s surface, having been adjusted for lens distortions, camera tilt and topographic relief. In this paper, we describe the different steps in the geometric correction process of APEX hyperspectral data, as applied in the Central Data Processing Center (CDPC at the Flemish Institute for Technological Research (VITO, Mol, Belgium. APEX ortho-images are generated through direct georeferencing of the raw images, thereby making use of sensor interior and exterior orientation data, boresight calibration data and elevation data. They can be referenced to any userspecified output projection system and can be resampled to any output pixel size.

  18. APEX - the Hyperspectral ESA Airborne Prism Experiment

    Directory of Open Access Journals (Sweden)

    Koen Meuleman

    2008-10-01

    Full Text Available The airborne ESA-APEX (Airborne Prism Experiment hyperspectral mission simulator is described with its distinct specifications to provide high quality remote sensing data. The concept of an automatic calibration, performed in the Calibration Home Base (CHB by using the Control Test Master (CTM, the In-Flight Calibration facility (IFC, quality flagging (QF and specific processing in a dedicated Processing and Archiving Facility (PAF, and vicarious calibration experiments are presented. A preview on major applications and the corresponding development efforts to provide scientific data products up to level 2/3 to the user is presented for limnology, vegetation, aerosols, general classification routines and rapid mapping tasks. BRDF (Bidirectional Reflectance Distribution Function issues are discussed and the spectral database SPECCHIO (Spectral Input/Output introduced. The optical performance as well as the dedicated software utilities make APEX a state-of-the-art hyperspectral sensor, capable of (a satisfying the needs of several research communities and (b helping the understanding of the Earth’s complex mechanisms.

  19. Creating Knockouts of Conserved Oligomeric Golgi Complex Subunits Using CRISPR-Mediated Gene Editing Paired with a Selection Strategy Based on Glycosylation Defects Associated with Impaired COG Complex Function.

    Science.gov (United States)

    Blackburn, Jessica Bailey; Lupashin, Vladimir V

    2016-01-01

    The conserved oligomeric Golgi (COG) complex is a key evolutionally conserved multisubunit protein machinery that regulates tethering and fusion of intra-Golgi transport vesicles. The Golgi apparatus specifically promotes sorting and complex glycosylation of glycoconjugates. Without proper glycosylation and processing, proteins and lipids will be mislocalized and/or have impaired function. The Golgi glycosylation machinery is kept in homeostasis by a careful balance of anterograde and retrograde trafficking to ensure proper localization of the glycosylation enzymes and their substrates. This balance, like other steps of membrane trafficking, is maintained by vesicle trafficking machinery that includes COPI vesicular coat proteins, SNAREs, Rabs, and both coiled-coil and multi-subunit vesicular tethers. The COG complex interacts with other membrane trafficking components and is essential for proper localization of Golgi glycosylation machinery. Here we describe using CRISPR-mediated gene editing coupled with a phenotype-based selection strategy directly linked to the COG complex's role in glycosylation homeostasis to obtain COG complex subunit knockouts (KOs). This has resulted in clonal KOs for each COG subunit in HEK293T cells and gives the ability to further probe the role of the COG complex in Golgi homeostasis. PMID:27632008

  20. Sarcocystis neurona infection in gamma interferon gene knockout (KO) mice: comparative infectivity of sporocysts in two strains of KO mice, effect of trypsin digestion on merozoite viability, and infectivity of bradyzoites to KO mice and cell culture.

    Science.gov (United States)

    Dubey, J P; Sundar, N; Kwok, O C H; Saville, W J A

    2013-09-01

    The protozoan Sarcocystis neurona is the primary cause of Equine Protozoal Myeloencephalitis (EPM). EPM or EPM-like illness has been reported in horses, sea otters, and several other mammals. The gamma interferon gene knockout (KO) mouse is often used as a model to study biology and discovery of new therapies against S. neurona because it is difficult to induce clinical EPM in other hosts, including horses. In the present study, infectivity of three life cycle stages (merozoites, bradyzoites, sporozoites) to KO mice and cell culture was studied. Two strains of KO mice (C57-black, and BALB/c-derived, referred here as black or white) were inoculated orally graded doses of S. neurona sporocysts; 12 sporocysts were infective to both strains of mice and all infected mice died or became ill within 70 days post-inoculation. Although there was no difference in infectivity of sporocysts to the two strains of KO mice, the disease was more severe in black mice. S. neurona bradyzoites were not infectious to KO mice and cell culture. S. neurona merozoites survived 120 min incubation in 0.25% trypsin, indicating that trypsin digestion can be used to recover S. neurona from tissues of acutely infected animals. PMID:23375195

  1. Cholesterol granuloma of the petrous apex: CT diagnosis

    Energy Technology Data Exchange (ETDEWEB)

    Lo, W.W.M.; Solti-Bohman, L.G.; Brackmann, D.E.; Gruskin, P.

    1984-12-01

    Cholesterol granuloma of the petrous apex is a readily recognizable and treatable entity that is more common than previously realized. Cholesterol granuloma grows slowly in the petrous apex as a mass lesion until it produces hearing loss, tinnitus, vertigo, and facial twitching. Twelve cases of cholesterol granuloma of the petrous apex are illustrated; ten of these analyzed in detail, especially with respect to CT findings. A sharply and smoothly marginated expansile lesion in the petrous apex, isodense with plain and nonenhancing on CT, is in all probability a cholesterol granuloma. Preoperative recognition by CT is important for planning proper treatment.

  2. Gene knockout mice establish a primary protective role for major histocompatibility complex class II-restricted responses in Chlamydia trachomatis genital tract infection.

    OpenAIRE

    Morrison, R P; Feilzer, K; Tumas, D B

    1995-01-01

    Mice with disrupted beta 2-microglobulin (beta 2m-/-), I-A (class II-/-), or CD4 (CD4-/-) genes were examined for their capacity to resolve Chlamydia trachomatis genital tract infection. C57BL/6 and beta 2m-/- mice resolved infection similarly and were culture negative by 4 to 5 weeks following infection. Conversely, major histocompatibility complex (MHC) class II-/- mice failed to resolve infection, and CD4-/- mice showed a significant delay (2 weeks). Secondary challenge of C57BL/6, beta 2m...

  3. A stringent dual control system overseeing transcription and activity of the Cre recombinase for the liver-specific conditional gene knock-out mouse model

    Institute of Scientific and Technical Information of China (English)

    Yu Wu; Yinghua He; Hongyu Zhang; Xinlan Dai; Xiaoyu Zhou; Jun Gu; Guan Wang; Jingde Zhu

    2008-01-01

    Liver cancer is one of the most threatening diseases in Chinese population. Just like in other tissues, tumor initiation and development in liver involve multiple steps of genetic and epigenetic alterations with several unknown details. However, unlike in other tissues, a tis- sue specific inducible Cre recombinase system that allows temporal and spatial deletion of a target DNA fragment is still not available for in vivo functional gene annotation in hepatocytes. In our pursuit to establish such a mouse model, we designed a dual inducible Cre transgene system and tested it in cultured cells. By combining a CCAAT/enhancer binding protein β (C/EBP β) promoter derived Tet-off expression system and the estrogen receptor (ER) mediated functional control, we show a desirable profile of both hepatocyte-specificity and regulability of the Cre expression in a series of critical assessments in the cell culture system, which provides confidence in continua- tion of our ongoing pursuit in mouse.

  4. Conditional Knockout Adam10 Gene of Neuronal Cell in Mouse Leads to Cortical Dysplasia%选择性敲除小鼠神经细胞 Adam10基因导致大脑皮质发育不良

    Institute of Scientific and Technical Information of China (English)

    王义辉; 李秀娟; 渠文生

    2015-01-01

    Objective: To investigate the role of Adam10 gene in mouse central nervous system (CNS) develop-ment. Methods: Gfapcre mice were mated to Adam10lox/lox mice using cre-loxp conditional gene knockout technique, and mice of conditional knockout Adam10 gene of neuronal cells(Gfapcre-Adam10lox/lox) were produced. At postnatal day of fourteen, mouse brain sections were prepared and HE staining was used to observe CNS development in Adam10 gene knockout mice. Results: By conditional knockout Adam10 gene of neuronal cell in mice (Gfapcre-Adam10lox/lox), mice could survive to about three weeks after birth. A tiny fraction of mice showed cortical defect,and developed cerebral cortex showed disorders of cortical lamination by HE staining. Conclusion: Adam10 gene plays an important role in CNS development. Conditional knockout Adam10 gene of neuronal cell in mouse leads to cortical defect or dysplasia.%目的:研究 Adam10基因在小鼠神经系统发育中的作用。方法:利用 Cre-loxp 转基因鼠技术,将 Gfapcre转基因鼠和 Adam10lox/lox 转基因鼠杂交,得到神经细胞特异性 Adam10基因敲除鼠(Gfapcre-Adam10lox/lox),在出生后第14天对大脑切片行 HE 染色,观察 Adam10基因敲除后神经系统发育情况。结果:选择性敲除小鼠神经细胞 Adam10基因(Gfapcre-Adam10lox/lox),小鼠可存活至出生后3周左右,部份小鼠大脑皮质单侧或双侧缺失,未出现皮质缺失的小鼠大脑皮质经 HE 染色提示出现层次结构紊乱。结论:Adam10基因在小鼠神经系统发育中具有重要作用,选择性敲除神经细胞 Adam10基因后导致小鼠大脑皮质缺失或层次结构紊乱。

  5. An Analysis of the Gene Expression in Mitf-m Knockout Mice%Mitf-m敲除小鼠的基因差异表达分析

    Institute of Scientific and Technical Information of China (English)

    王克双; 王晓东; 任丽丽; 陈磊; 塞娜; 郭维维; 杨仕明

    2016-01-01

    目的 使用RNA-Seq技术分析Mitf-m敲除鼠与野生鼠的血管纹转录组,研究Mitf-m基因敲除后的差异表达基因及相关改变的分子机制.方法 分别取Mitf-m敲除鼠(Mitfm’△M/mi-△M;MM组)与野生鼠(Mitf-m+/+;WW组)的血管纹进行总RNA提取;制备cDNA文库,用Illumina HiSeq 2000测序系统行高通量测序.利用软件Tophat 2.2.0将clean reads比对到参考基因组;分别用软件RSeQS-2.3.2和cufflinks来检测测序结果的均一性和基因表达水平.使用edgeR和DESeq程序筛选差异表达基因(differential expressed genes,DEGs);选择下调DEGs,用KOBAS2.0进行基因本体(gene ontology,GO)和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)代谢途径的富集分析.结果 MM组与WW组在参考基因组上mapping的clean reads数分别为42463888和3971 8542,分别有88.7%和87.4%的reads是唯一映射,说明RNA-Seq结果可靠.DEGs筛查结果表明,相对于WW组,Mitf-m敲除鼠血管纹有45个DEGs,上调表达基因有7个,下调表达基因有38个.GO富集分析发现下调表达基因与黑色素的合成和代谢过程、色素沉着有关,值得关注的是与内向整流钾离子通道Kcnj 10和Kcnj13相关.KEGG富集分析显示下调表达基因主要富集于黑色素生成通路.结论 RNA-Seq分析拓展了Mitf-m基因调控网,为探究Mitf-m突变所致听力-色素综合征的分子机制及特异性研究Mitf不同转录子的功能奠定了基础.

  6. Mitf-m敲除小鼠的基因差异表达分析%An Analysis of the Gene Expression in Mitf-m Knockout Mice

    Institute of Scientific and Technical Information of China (English)

    王克双; 王晓东; 任丽丽; 陈磊; 塞娜; 郭维维; 杨仕明

    2016-01-01

    目的 使用RNA-Seq技术分析Mitf-m敲除鼠与野生鼠的血管纹转录组,研究Mitf-m基因敲除后的差异表达基因及相关改变的分子机制.方法 分别取Mitf-m敲除鼠(Mitfm’△M/mi-△M;MM组)与野生鼠(Mitf-m+/+;WW组)的血管纹进行总RNA提取;制备cDNA文库,用Illumina HiSeq 2000测序系统行高通量测序.利用软件Tophat 2.2.0将clean reads比对到参考基因组;分别用软件RSeQS-2.3.2和cufflinks来检测测序结果的均一性和基因表达水平.使用edgeR和DESeq程序筛选差异表达基因(differential expressed genes,DEGs);选择下调DEGs,用KOBAS2.0进行基因本体(gene ontology,GO)和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)代谢途径的富集分析.结果 MM组与WW组在参考基因组上mapping的clean reads数分别为42463888和3971 8542,分别有88.7%和87.4%的reads是唯一映射,说明RNA-Seq结果可靠.DEGs筛查结果表明,相对于WW组,Mitf-m敲除鼠血管纹有45个DEGs,上调表达基因有7个,下调表达基因有38个.GO富集分析发现下调表达基因与黑色素的合成和代谢过程、色素沉着有关,值得关注的是与内向整流钾离子通道Kcnj 10和Kcnj13相关.KEGG富集分析显示下调表达基因主要富集于黑色素生成通路.结论 RNA-Seq分析拓展了Mitf-m基因调控网,为探究Mitf-m突变所致听力-色素综合征的分子机制及特异性研究Mitf不同转录子的功能奠定了基础.

  7. Investigation on the mechanism of descended reproductive function in FMR1 gene knockout male mice%FMR1基因敲除雄鼠生殖功能下降的机制探讨

    Institute of Scientific and Technical Information of China (English)

    叶球仙; 杨洁; 罗虹; 陈盛强; 黄晓虹; 甘婷; 陈燕; 肖国宏

    2013-01-01

    Objective To investigate the mechanism of descended reproductive function induced by FMR1 gene knockout in male FMR1 gene knockout mice (KO) and male wild mice (WT).Methods The morphology of the testis of 10 weeks old KO and WT male mice were observed by HE staining.The mean optical density (MOD) of the expressions of neuropeptide Y (NPY) and gamma aminobutyricacid (GABA) in hypothalamus were tested and analyzed by immunohistochemistry and Image Pro Plus 6.0.Serum NPY concentrations were measured by ELISA.Results No obvious pathological differences in testis were found between KO and WT mice.The MOD of NPY in hypothalamus and the expression of NPY in serum in KO mice were weaken than those in WT mice,respectively [0.27 ± 0.031 vs.0.31 ± 0.031,P < 0.05; (179.21 ± 50.773)ng/L vs.(225.24 ± 52.293) ng/L,P <0.05].There was no significant difference in the MOD of GABA in hypothalamus between KO and WT mice (0.29 ±0.017 vs.0.28 ± 0.009,P > 0.05).Conclusions FMR1 gene may down-regulate the reproductive function of male mice through reducing the expression of NPY.%目的:利用FMR1基因敲除鼠(KO)和野生型FVB鼠(WT),研究FMR1基因敲除引起雄鼠生殖功能下降的可能机制.方法:随机取10周的KO及WT雄鼠,HE染色观察睾丸形态;免疫组化及Image ProPlus 6.0测定分析下丘脑神经肽Y(NPY)及γ-氨基丁酸(GABA)表达的平均光密度值;ELISA测定血清NPY水平.结果:KO、WT小鼠睾丸形态未见明显不同;KO雄鼠下丘脑NPY的表达弱于WT雄鼠(0.27±0.031,0.31±0.031,P<0.05);KO雄鼠血清NPY的表达弱于WT雄鼠(179.21±50.773,225.24±52.293,P< 0.05);两种基因型小鼠下丘脑GABA的表达差异无统计学意义(0.29±0.017,0.28±0.009,P> 0.05).结论:FMR1基因可能是通过下调NPY的表达来降低雄鼠生殖功能的.

  8. Atmospheric correction of APEX hyperspectral data

    Directory of Open Access Journals (Sweden)

    Sterckx Sindy

    2016-03-01

    Full Text Available Atmospheric correction plays a crucial role among the processing steps applied to remotely sensed hyperspectral data. Atmospheric correction comprises a group of procedures needed to remove atmospheric effects from observed spectra, i.e. the transformation from at-sensor radiances to at-surface radiances or reflectances. In this paper we present the different steps in the atmospheric correction process for APEX hyperspectral data as applied by the Central Data Processing Center (CDPC at the Flemish Institute for Technological Research (VITO, Mol, Belgium. The MODerate resolution atmospheric TRANsmission program (MODTRAN is used to determine the source of radiation and for applying the actual atmospheric correction. As part of the overall correction process, supporting algorithms are provided in order to derive MODTRAN configuration parameters and to account for specific effects, e.g. correction for adjacency effects, haze and shadow correction, and topographic BRDF correction. The methods and theory underlying these corrections and an example of an application are presented.

  9. APEX: A Prime EXperiment at Jefferson Lab

    Directory of Open Access Journals (Sweden)

    Beacham James

    2014-06-01

    Full Text Available APEX is an experiment at Thomas Jefferson National Accelerator Facility (JLab in Virginia, USA, that searches for a new gauge boson (A′ with sub-GeV mass and coupling to ordinary matter of g′ ∼ (10−6 − 10−2e. Electrons impinge upon a fixed target of high-Z material. An A′ is produced via a process analogous to photon bremsstrahlung, decaying to an e+e− pair. A test run was held in July of 2010, covering mA′ = 175 to 250 MeV and couplings g′/e > 10−3. A full run is approved and will cover mA′ ∼65 to 525 MeV and g′/e > 2.3 × 10−4.

  10. APEX: A Prime EXperiment at Jefferson Lab

    Directory of Open Access Journals (Sweden)

    Beacham James

    2015-01-01

    Full Text Available APEX is an experiment at Thomas Jefferson National Accelerator Facility (JLab in Virginia, USA, that searches for a new gauge boson (A′ with sub-GeV mass and coupling to ordinary matter of g′ ∼ (10−6 − 10−2e. Electrons impinge upon a fixed target of high-Z material. An A′ is produced via a process analogous to photon bremsstrahlung, decaying to an e+e− pair. A test run was held in July of 2010, covering mA′ = 175 to 250 MeV and couplings g′/e > 10−3. A full run is approved and will cover mA′ ∼ 65 to 525 MeV and g′/e > 2.3 × 10−4, and is expected to occur sometime in 2016 or 2017.

  11. Towards Accurate Application Characterization for Exascale (APEX)

    Energy Technology Data Exchange (ETDEWEB)

    Hammond, Simon David [Sandia National Laboratories (SNL-NM), Albuquerque, NM (United States)

    2015-09-01

    Sandia National Laboratories has been engaged in hardware and software codesign activities for a number of years, indeed, it might be argued that prototyping of clusters as far back as the CPLANT machines and many large capability resources including ASCI Red and RedStorm were examples of codesigned solutions. As the research supporting our codesign activities has moved closer to investigating on-node runtime behavior a nature hunger has grown for detailed analysis of both hardware and algorithm performance from the perspective of low-level operations. The Application Characterization for Exascale (APEX) LDRD was a project concieved of addressing some of these concerns. Primarily the research was to intended to focus on generating accurate and reproducible low-level performance metrics using tools that could scale to production-class code bases. Along side this research was an advocacy and analysis role associated with evaluating tools for production use, working with leading industry vendors to develop and refine solutions required by our code teams and to directly engage with production code developers to form a context for the application analysis and a bridge to the research community within Sandia. On each of these accounts significant progress has been made, particularly, as this report will cover, in the low-level analysis of operations for important classes of algorithms. This report summarizes the development of a collection of tools under the APEX research program and leaves to other SAND and L2 milestone reports the description of codesign progress with Sandia’s production users/developers.

  12. KnockoutJS web development

    CERN Document Server

    Farrar, John

    2015-01-01

    This book is for web developers and designers who work with HTML and JavaScript to help them manage data and interactivity with data using KnockoutJS. Knowledge about jQuery will be useful but is not necessary.

  13. Construction of H22 GP73 knockout gene stable strain using CRISPR/Cas9 gene editing system and identification of functions%利用CRISPR/Cas9系统构建H22细胞GP73基因敲除稳定株及功能鉴定

    Institute of Scientific and Technical Information of China (English)

    陈健康; 魏从文; 梁慧; 王贝晗; 钟辉; 杨晓莉

    2016-01-01

    目的:利用CRISPR/Cas9基因编辑系统敲除小鼠源肝癌细胞H22中的GP73基因,构建H22细胞GP73基因敲除的稳定细胞株。方法根据CRISPR/Cas9靶点设计原则,设计2条特异性识别GP73基因启动子的上下游sgRNA,利用载体pX459质粒,构建2对重组真核表达质粒。经酶切和测序鉴定后,将重组质粒转染至H22细胞内,使用嘌罗霉素加压筛选稳定敲除GP73的H22细胞株,利用免疫印迹检测重组质粒对内源GP73的敲除效果。MTT实验检测GP73被敲除后对细胞增殖能力的影响,再利用划痕实验检测细胞的迁移能力。结果免疫印迹结果说明敲除GP73基因的小鼠源H22细胞株内无GP73蛋白的表达;并且GP73被敲除后,H22细胞的增殖能力和迁移能力减慢。结论通过CRISPR/Cas9系统获得了靶向GP73基因的重组质粒,并且筛选出了稳定干扰GP73表达的细胞株,从而为探讨GP73在肝癌发生中的作用奠定基础。%Objective To knock out the GP73 gene in H22 cells originating in mice using CRISPR/Cas9 gene editing system and construct H22 GP73 gene knockout stable strain for identification of its functions .Methods Two pairs of sgRNAs that could specifically identify the upstream and downstream of GP 73 gene first promoter were designed before a recombinant eukaryotic expressional plasmid was constructed using pX 459 .After enzyme digestion and sequencing , two pairs of recombinant plasmids were co-transfected into H22 cells before puromycin was used to screen positive cells and monoclonal cells which stably knocked out GP 73 gene were developed .The knockout effect was measured by Western blotting.Cell Titer 96® AQueous One Solution Assay was used to detect the effect on cell reproductive capacity when the GP73 was knocked out .The transferability was detected through wound healing test .Results The result of Western blotting suggested that GP73 protein was undetected in the construction of H22

  14. FMR1基因敲除对雄性小鼠生殖功能的影响%The influence of FMR1 gene knockout on the reproduction of male mice

    Institute of Scientific and Technical Information of China (English)

    祝亚桥; 周兴; 陈盛强

    2012-01-01

    Objective To investigate the influence of fragile x mental retardation-1 (FMR1) gene on the spermatogenesis and reproduction of male mice. Methods FMR1 knockout (KO) male mice and wild type (WT) male mice were mated with wt female mice. The number of litters, pregnancy rate and male mice having offsprings were counted. Serum T, FSH and LH concentrations were also measured. The density, mortality and morphology of the left cauda epididymis sperms were analyzed, HE staining was performed on the right side. Results The pregnance rate of wt female mice mated with FMR1K0 males was significantly lower than the control group (41.7% vs. 87. 5% , P 0.05). Male fertility showed that 41.7% of KO mice had pups, whereas 91.7% of the mice had pups in the control group (P 0.05).两组小鼠血清T,FSH,LH浓度无统计学差异.KO组的睾丸附睾病理切片与WT组比较未见明显异常,其精子活率及各种畸形率与WT小鼠比较均没有统计学差异(P>0.05).结论:可以推测FMR1基因对雄性生殖系统发育有一定的影响,Fmr1基因的缺失降低了雄性小鼠生育率,但对精子生成、畸形率等未见明显影响,其对雄性生殖系统影响机制还有待进一步的实验研究.

  15. INDUCTION OF MAMMARY GLAND DEVELOPMENT IN ESTROGEN RECEPTOR-ALPHA KNOCKOUT MICE

    Science.gov (United States)

    Mammary glands from the estrogen receptor knockout ( ERKO) mouse do not undergo ductal morphogenesis or alveolar development. Disrupted Er signaling may result in reduced estrogen-responsive gene products in the mammary gland or reduced mammotropic hormones that contribute t...

  16. Plastid osmotic stress influences cell differentiation at the plant shoot apex.

    Science.gov (United States)

    Wilson, Margaret E; Mixdorf, Matthew; Berg, R Howard; Haswell, Elizabeth S

    2016-09-15

    The balance between proliferation and differentiation in the plant shoot apical meristem is controlled by regulatory loops involving the phytohormone cytokinin and stem cell identity genes. Concurrently, cellular differentiation in the developing shoot is coordinated with the environmental and developmental status of plastids within those cells. Here, we employ an Arabidopsis thaliana mutant exhibiting constitutive plastid osmotic stress to investigate the molecular and genetic pathways connecting plastid osmotic stress with cell differentiation at the shoot apex. msl2 msl3 mutants exhibit dramatically enlarged and deformed plastids in the shoot apical meristem, and develop a mass of callus tissue at the shoot apex. Callus production in this mutant requires the cytokinin receptor AHK2 and is characterized by increased cytokinin levels, downregulation of cytokinin signaling inhibitors ARR7 and ARR15, and induction of the stem cell identity gene WUSCHEL Furthermore, plastid stress-induced apical callus production requires elevated plastidic reactive oxygen species, ABA biosynthesis, the retrograde signaling protein GUN1, and ABI4. These results are consistent with a model wherein the cytokinin/WUS pathway and retrograde signaling control cell differentiation at the shoot apex. PMID:27510974

  17. Root Apex Transition Zone as Oscillatory Zone

    Directory of Open Access Journals (Sweden)

    Frantisek eBaluska

    2013-10-01

    Full Text Available Root apex of higher plants shows very high sensitivity to environmental stimuli. The root cap acts as the most prominent plant sensory organ; sensing diverse physical parameters such as gravity, light, humidity, oxygen and critical inorganic nutrients. However, the motoric responses to these stimuli are accomplished in the elongation region. This spatial discrepancy was solved when we have discovered and characterized the transition zone which is interpolated between the apical meristem and the subapical elongation zone. Cells of this zone are very active in the cytoskeletal rearrangements, endocytosis and endocytic vesicle recycling, as well as in electric activities. Here we discuss the oscillatory nature of the transition zone which, together with several other features of this zone, suggest that it acts as some kind of command centre. In accordance with the early proposal of Charles and Francis Darwins, cells of this root zone receive sensory information from the root cap and instruct the motoric responses of cells in the elongation zone.

  18. Resection of the Tooth Apex with Diode Laser

    Directory of Open Access Journals (Sweden)

    Uzunov Tz.

    2014-06-01

    Full Text Available An “in vitro” experimental study has been carried out on 70 extracted teeth. A laser resection of the root apex has been carried out with diode laser beam with a wavelength of - 810 ± 10 nm. Sequentially a radiation with increasing power has been applied, as follows: 1,3 W, 2W, 3W, 4W, 5W, 6W, 7W, in electro surgery mode. Successful resection of the tooth apex has been performed at: 3W; 4W; 5W; 6W and 7W power. It was established that when laser resected the tooth apex carbonizes.

  19. Rare complete knockouts in humans: population distribution and significant role in autism spectrum disorders

    Science.gov (United States)

    Lim, Elaine T.; Raychaudhuri, Soumya; Sanders, Stephan J.; Stevens, Christine; Sabo, Aniko; MacArthur, Daniel G.; Neale, Benjamin M.; Kirby, Andrew; Ruderfer, Douglas M.; Fromer, Menachem; Lek, Monkol; Liu, Li; Flannick, Jason; Ripke, Stephan; Nagaswamy, Uma; Muzny, Donna; Reid, Jeffrey G.; Hawes, Alicia; Newsham, Irene; Wu, Yuanqing; Lewis, Lora; Dinh, Huyen; Gross, Shannon; Wang, Li-San; Lin, Chiao-Feng; Valladares, Otto; Gabriel, Stacey B.; dePristo, Mark; Altshuler, David M.; Purcell, Shaun M.; State, Matthew W.; Boerwinkle, Eric; Buxbaum, Joseph D.; Cook, Edwin H.; Gibbs, Richard A.; Schellenberg, Gerard D.; Sutcliffe, James S.; Devlin, Bernie; Roeder, Kathryn; Daly, Mark J.

    2013-01-01

    SUMMARY To characterize the role of rare complete human knockouts in autism spectrum disorders (ASD), we identify genes with homozygous or compound heterozygous loss-of-function (LoF) variants (defined as nonsense and essential splice sites) from exome sequencing of 933 cases and 869 controls. We identify a two-fold increase in complete knockouts of autosomal genes with low rates of LoF variation (≤5% frequency) in cases and estimate a 3% contribution to ASD risk by these events, confirming this observation in an independent set of 563 probands and 4,605 controls. Outside the pseudo-autosomal regions on the X-chromosome, we similarly observe a significant 1.5-fold increase in rare hemizygous knockouts in males, contributing to another 2% of ASDs in males. Taken together these results provide compelling evidence that rare autosomal and X-chromosome complete gene knockouts are important inherited risk factors for ASD. PMID:23352160

  20. Surface Free Energy Determination of APEX Photosensitive Glass

    Directory of Open Access Journals (Sweden)

    William R. Gaillard

    2016-02-01

    Full Text Available Surface free energy (SFE plays an important role in microfluidic device operation. Photosensitive glasses such as APEX offer numerous advantages over traditional glasses for microfluidics, yet the SFE for APEX has not been previously reported. We calculate SFE with the Owens/Wendt geometric method by using contact angles measured with the Sessile drop technique. While the total SFE for APEX is found to be similar to traditional microstructurable glasses, the polar component is lower, which is likely attributable to composition. The SFE was modified at each stage of device fabrication, but the SFE of the stock and fully processed glass was found to be approximately the same at a value of 51 mJ·m−2. APEX exhibited inconsistent wetting behavior attributable to an inhomogeneous surface chemical composition. Means to produce more consistent wetting of photosensitive glass for microfluidic applications are discussed.

  1. Apex Organizations and the Growth of Microfinance in Bolivia

    OpenAIRE

    Sergio Navajas; Mark Schreiner

    2001-01-01

    Bolivia has the most advanced microfinance sector in Latin America and has been a model worldwide. Apex organizations--second-tier wholesaling mechanisms that lend and offer non- financial assistance to retailing microfinance organizations--have not been responsible for this success. Former and current Bolivian apex organizations have engaged in little market development. Some have provided some liquidity to microfinance organizations, but they have not played an indispensable role in the dev...

  2. Pleiotropic effects in Eya3 knockout mice

    Directory of Open Access Journals (Sweden)

    Naton Beatrix

    2008-12-01

    Full Text Available Abstract Background In Drosophila, mutations in the gene eyes absent (eya lead to severe defects in eye development. The functions of its mammalian orthologs Eya1-4 are only partially understood and no mouse model exists for Eya3. Therefore, we characterized the phenotype of a new Eya3 knockout mouse mutant. Results Expression analysis of Eya3 by in-situ hybridizations and β-Gal-staining of Eya3 mutant mice revealed abundant expression of the gene throughout development, e.g. in brain, eyes, heart, somites and limbs suggesting pleiotropic effects of the mutated gene. A similar complex expression pattern was observed also in zebrafish embryos. The phenotype of young adult Eya3 mouse mutants was systematically analyzed within the German Mouse Clinic. There was no obvious defect in the eyes, ears and kidneys of Eya3 mutant mice. Homozygous mutants displayed decreased bone mineral content and shorter body length. In the lung, the tidal volume at rest was decreased, and electrocardiography showed increased JT- and PQ intervals as well as decreased QRS amplitude. Behavioral analysis of the mutants demonstrated a mild increase in exploratory behavior, but decreased locomotor activity and reduced muscle strength. Analysis of differential gene expression revealed 110 regulated genes in heart and brain. Using real-time PCR, we confirmed Nup155 being down regulated in both organs. Conclusion The loss of Eya3 in the mouse has no apparent effect on eye development. The wide-spread expression of Eya3 in mouse and zebrafish embryos is in contrast to the restricted expression pattern in Xenopus embryos. The loss of Eya3 in mice leads to a broad spectrum of minor physiological changes. Among them, the mutant mice move less than the wild-type mice and, together with the effects on respiratory, muscle and heart function, the mutation might lead to more severe effects when the mice become older. Therefore, future investigations of Eya3 function should focus on

  3. Establishment of murine Smad5 double knockout ES cells and the studies on their properties

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Smad5 is an intracellular transducer of TGF-b signals. Targeteddisruption of murine Smad5 gene resulted in embryonic lethal. To study the function of Smad5 in organgenesis, we generated Smad5 double knockout ES cells by homologous recombination. We deleted the neo gene of the Smad5 targeted ES cells using Cre-LoxP system. Smad5 double knockout ES cells were obtained by transfecting the targeted ES cells using the same targeting construct. The results of chimeric study showed that Smad5 might play an important role during the development of heart and neural tube. Smad5 double knockout ES cells formed teratoma when injected subcutaneously into nude mice. They differentiated into several types of cells, including neural cells, muscle cells, chondrocytes, endothelial cells and glandaceous cells. Smad5 double knockout ES cells are useful for studying the function of Smad5 mediated TGF- b during the organgenesis and the in vitro differentiation of ES cells.

  4. [An operative case of cholesterol granuloma of the petrous apex].

    Science.gov (United States)

    Saino, M; Kayama, T; Kuroki, A; Siraisi, Y; Sato, K; Nakai, O

    1996-11-01

    A 59-year-old man presented with a rare cholesterol granuloma of the petrous apex manifesting as headache, left facial dysesthesia, diplopia, left hearing impairment, and left tinnitus. Neurological examination revealed dysesthesia of territory in all divisions of the left trigeminal nerve, left incomplete abducens nerve palsy, left mixed hearing impairment, and left tinnitus. Plain CT scan showed a smoothly marginated mass involving the left petrous apex. The mass was isodense with the brain parenchyma and not enhanced by contrast medium. The mass appeared heterogeneously slightly hyperintense on the T1-weighted MR image and homogeneously hyperintense on the T2-weighted MR image except for the peripheral portion. The mass was not enhanced after intravenous gadolinium DTPA administration. Surgery via a petrosal approach totally removed the mass in the intracranial, extradural space. Histological examination showed typical features of cholesterol granuloma, with cholesterin clefts, hemosiderin deposits, and erythrocytes in non-specific granulation tissue. Cholesterol granuloma most commonly occurs in the middle ear cavity, and rarely in the petrous apex. The characteristic hyperintense appearance of cholesterol granuloma on T1- and T2-weighted MR images is very useful for differentiation from other lesions of the petrous apex and the cerebellopontine angle such as cholesteatoma, mucocele, chordoma, and meningioma. Solid cholesterol granuloma of the petrous apex should be treated by total removal via craniotomy, not by drainage which is commonly performed by otorhinologists. PMID:8934474

  5. Production and Characterization of Anti-PrP Polyclonal Antibody by PRNP Gene Knockout Goat%利用PRNP-/-羊制备朊蛋白多克隆抗体及其特性分析

    Institute of Scientific and Technical Information of China (English)

    王学斌; 俞慧清; 吴晓东; 徐旭俊; 陈建泉; 成国祥

    2012-01-01

    [Objective] The study was conducted in order to produce and characterize anti-PrP polyclonal antibodies by prion protein knockout (PRNP'') goat. [Method] Using polymerase chain reaction (PCR) method, the DNA sequence encoding the goat mature PrP (Prion Protein) was amplified from goat DNA. It was then cloned into the vector pET-22b to express recombinant PrP protein. After purification, the recombinant PrP was used as antigen to immunize PRNP'1' goat for producing anti-PrP antibodies. ELISA and Western blot were conducted to characterize the titer and specificity of polyclonal antibodies. [Result] A large scale of anti-PrP polyclonal antibodies was successfully produced by PRNP'' goat. ELISA assay revealed that the titer of the polyclonal antibodies against PrP was as high as 1 : 25 600. Western blot test showed that the antibody was not only able to react with the native prion proteins (PrPc) of mouse, cattle and goat brain, but also could combine with misfolded prion proteins (PrPSc) from mouse brain. [Conclusion] Ablation of the prion protein(PrP) gene in goat facilitates the production of a large scale anti-PrP polyclonal antibodies, the produced antibody can be widely used in detecting PrPc and PrPSc derived from a variety of animals.%[目的]利用朊蛋白双基因敲除(PRNP-/-)羊制备朊蛋白多克隆抗体,并对其特性进行分析.[方法]构建山羊朊蛋白( PrP)原核表达载体,转入大肠杆菌并诱导表达,纯化获得羊PrP;将获得的PrP免疫pRNP-/-山羊,制备朊蛋白特异性的多克隆抗体;并对获得的朊蛋白多克隆抗体进行ELISA及Western-blot检测.[结果]获得了大量的朊蛋白特异性抗血清,间接ELASA检测抗血清中朊蛋白多克隆抗体的效价为25600;Western-blot 检测显示所制备抗体不仅可以识别鼠、牛、羊脑组织内源性朊蛋白,而且能识别鼠脑组织内朊病毒.[结论]PRNP-/-转基因山羊可用于制备大量高亲和力朊蛋白多克隆抗体,获得的

  6. Effects of Eaf2 gene knockout on cataract induced by ultraviolet irradiation in mice%Eaf2基因敲除对紫外线诱导的鼠白内障形成的影响

    Institute of Scientific and Technical Information of China (English)

    姜艳华; 张劲松

    2016-01-01

    目的:分析Eaf2基因敲除对紫外线诱导的小鼠白内障形成的影响。方法:将15只野生型( WT)小鼠作为对照组,10只Eaf2基因敲除( Eaf2 KO)小鼠作为实验组。两组均取14周龄左右的小鼠作为研究对象。进一步分组为:WT-nonUV、WT-UV、Eaf2 KO-nonUV、Eaf2 KO-UV,共4组。使用裂隙灯显微镜观察小鼠白内障程度,利用晶状体混浊分类系统Ⅱ( LOCSⅡ)对小鼠白内障进行分级。然后断颈处死小鼠,摘取晶状体进行暗视野显微镜照相,利用Image J软件对晶状体混浊程度进行分析,并将各测量结果进行统计学处理。结果:裂隙灯显微镜和暗视野显微镜的结果一致:WT-UV组及 Eaf2 KO-UV 组晶状体混浊程度明显高于 WT-nonUV组及Eaf2 KO-nonUV组,其中WT-UV组明显高于Eaf2 KO-UV组,均具有统计学差异(P<0.05)。结论:紫外线辐射能够导致小鼠白内障的形成,Eaf2蛋白质具有促进紫外线所致的小鼠白内障形成的作用。%Abstract•AIM:To evaluate the effects of Eaf2 gene knockout on cataract in mice induced by ultraviolet irradiation.•METHODS:Fifteen wild type mice were used as the control group, and 10 Eaf2 KO mice were used as the experimental group.The 14-week mice were taken as the research objects in the two groups. So the subgroups were:WT -nonUV, WT -UV, Eaf2 KO-nonUV and Eaf2 KO-UV, a total of 4 groups.Observe the lens of mice in vivo with slit lamp microscope, grade the lens opacity with Lens Opacities Classification System II ( LOCSII ) . Then the mice were sacrificed by breaking the neck, the lens were removed and were observed by dark field microscopy. According to the captured images, the proportion of cataract region was analyzed using Image J software.The data of the two groups were statistically analyzed.•RESULTS: The results detected by the two methods were similar.In WT-UV group and Eaf2 KO-UV group, the degree of lens

  7. The APEX-SZ instrument: Development, performance and observations

    Science.gov (United States)

    Schwan, Daniel

    This thesis describes the development, design, instrument performance and observations of the APEX-SZ receiver. APEX-SZ is a millimeter-wave cryogenic receiver designed to observe galaxy clusters via the Sunyaev-Zel'dovich effect from the 12 m APEX telescope on the Atacama plateau in Chile. The receiver consists of a focal plane of 280 superconducting transition edge sensor (TES) bolometers cooled to 280 mK via a three stage He sorption fridge and mechanical pulse tube cooler. The detectors are instrumented with a frequency-domain multiplexed readout system. A horn array, 4 K lenses and tertiary mirrors couple the TES bolometers to the telescope. APEX-SZ observes in a single frequency band at 150 GHz with 59" angular resolution and a 22' field-of-view, well suited for cluster mapping. The receiver represents a leap forward in millimeter wave detector technology. Initial observations with APEX-SZ were early demonstrations of a fielded imaging TES array, frequency multiplexed readout and cryocooled bolometer receiver. These technologies have already been applied to receivers with thousands of detectors. APEX-SZ typically observes with 180 optically active detectors with an optical efficiency of 0.33 and median NET per channel of 860 muKCMB s . The per channel sensitivity is limited by a bandwidth 40% narrower than expected, and cavity absorption efficiency of 0.74. New detector wafers with wider bandwidth have been fabricated and tested at the telescope on a limited run last spring. With improved bandwidth and lower thermal conductivity, newly fabricated wafers can achieve background noise limited performance. We plan to upgrade the focal plane with a full array of new wafers in December 2009. The receiver has successfully completed 9 weeks of science observation since its installation in April 2007. We have made over a dozen SZE maps from targeted observations of known clusters. We present SZE temperature maps of the Bullet cluster (1E 0657--56) and Abell 2163.

  8. Human Genetic Disorders and Knockout Mice Deficient in Glycosaminoglycan

    Directory of Open Access Journals (Sweden)

    Shuji Mizumoto

    2014-01-01

    Full Text Available Glycosaminoglycans (GAGs are constructed through the stepwise addition of respective monosaccharides by various glycosyltransferases and maturated by epimerases and sulfotransferases. The structural diversity of GAG polysaccharides, including their sulfation patterns and sequential arrangements, is essential for a wide range of biological activities such as cell signaling, cell proliferation, tissue morphogenesis, and interactions with various growth factors. Studies using knockout mice of enzymes responsible for the biosynthesis of the GAG side chains of proteoglycans have revealed their physiological functions. Furthermore, mutations in the human genes encoding glycosyltransferases, sulfotransferases, and related enzymes responsible for the biosynthesis of GAGs cause a number of genetic disorders including chondrodysplasia, spondyloepiphyseal dysplasia, and Ehlers-Danlos syndromes. This review focused on the increasing number of glycobiological studies on knockout mice and genetic diseases caused by disturbances in the biosynthetic enzymes for GAGs.

  9. Study of foramen openings and their concurrence with root apexes

    Directory of Open Access Journals (Sweden)

    Sandra Maria Alves SAYÃO MAIA

    2005-05-01

    Full Text Available The present study is aimed at evaluating the anatomic concurrencebetween foramen openings and root apexes of 247 upper and lowerpermanent human molar canals, the distance between these structures,the instrument that best fits into the root canal, as well as the direction of foramen deviation. Sixty-four of the canals were partially impenetrable and were discarded. The findings showed that 39.9% of the root canals studied had their apical foramen concurrent with their root apexes and 60.1% did not. Clinicians must be made aware of this important anatomical detail, which could be indispensable for successful endodontic treatment.

  10. Pax-8基因敲除小鼠心脏中NR4A1基因表达上调%Up-regulation of NR4A1 gene expression in Pax-8 gene knockout mice myocardium

    Institute of Scientific and Technical Information of China (English)

    黄晓燕; 高瞻; 周希; 梁万前; 陈锡文; 陈通克; 杨德业

    2011-01-01

    Objective: To find the possible genic pathway for the anti-apoptosis function of the ventricular septal defeet related to Pax-8. Method: The total RNA derived from the heart of Pax-8 KO-/- and Pax-8 KO+/- mice was extracted. Mouse genome DNA microarray was used to detect the differential expression level between the Pax-8 KO-/- and the Pax-8 KO+/- mice heart. The candidate genes were confirmed by RT-PCR and real time RT-PCR assay. Results: Microarray results showed that 25 genes were down-regulated and 17 were up-regulated in the Pax-8 KO-/- mice compared to the Pax-8 KO+/- mice. Nuclear receptor subfamily 4, group A, member 1 (NR4A1)was proved to be up-regulated by RT-PCR in the Pax-8 KO-/- mice heart. Real time RTPCR results revealed that NR4A1 in the Pax-8 KO-/- mice was 1.53 and 4.79 fold as much as in the Pax-8 KO+/- and the Pax-8+/+ mice (P<0.01). Conclusion: The NR4A1 gene is one of the downstream genes of the Pax-8 and probably evolved and played an important role in the mechanism of ventricular septum defect.%目的:寻找先天性心脏病室间隔缺损相关基因--转录因子Pax-8保护细胞凋亡的途径.方法:提取Pax-8基因敲除小鼠纯合子(Pax-8 KO-/-)和杂合子(Pax-8 KO+/-)的心脏总RNA,利用小鼠基因的基因芯片检测两组小鼠基因表达水平,找出差异表达的基因,并经半定量RT-PCR和荧光实时定量PCR技术初步筛选出转录因子Pax-8的下游基因.结果:基因芯片检测发现,与Pax-8 KO+/-组相比,在Pax-8 KO-/-小鼠中有25个基因表达下调,17个基因表达上调.用半定量RT-PCR验证发现,nuclear receptor sub-family4,group A,member 1(NR4A1)基因在Pax-8 KO-/-组上调.定量RT-PCR亦证实在Pax-8 KO-/-组NR4A1基因的表达水平较Pax-8 KO+/-组及Pax-8+/+(野生型)组分别上调1.53倍和4.79倍(P<0.01).结论:NR4A1基因受Pax-8基因调控,在Pax-8保护细胞凋亡途径中发挥作用.

  11. Nitric Oxide-Mediated Maize Root Apex Responses to Nitrate are Regulated by Auxin and Strigolactones

    Directory of Open Access Journals (Sweden)

    Alessandro eManoli

    2016-01-01

    Full Text Available Nitrate (NO3- is a key element for crop production but its levels in agricultural soils are limited. Plants have developed mechanisms to cope with these NO3- fluctuations based on sensing nitrate at the root apex. Particularly, the transition zone (TZ of root apex has been suggested as a signalling-response zone. This study dissects cellular and molecular mechanisms underlying NO3- resupply effects on primary root growth in maize, confirming nitric oxide (NO as a putative modulator. Nitrate restoration induced primary root elongation within the first 2 h, corresponding to a stimulation of cell elongation at the basal border of the TZ. Xyloglucans (XGs immunolocalization together with Brefeldin A applications demonstrated that nitrate resupply induces XG accumulation. This effect was blocked by cPTIO (NO scavenger. Transcriptional analysis of ZmXET1 confirmed the stimulatory effect of nitrate on XGs accumulation in cells of the TZ. Immunolocalization analyses revealed a positive effect of nitrate resupply on auxin and PIN1 accumulation, but a transcriptional regulation of auxin biosynthesis/transport/signalling genes was excluded. Short-term nitrate treatment repressed the transcription of genes involved in strigolactones (SLs biosynthesis and transport, mainly in the TZ. Enhancement of carotenoid cleavage dioxygenases (CCDs transcription in presence of cPTIO indicated endogenous NO as a negative modulator of CCDs activity. Finally, treatment with the SLs-biosynthesis inhibitor (TIS108 restored the root growth in the nitrate-starved seedlings. Present report suggests that the NO-mediated root apex responses to nitrate are accomplished in cells of the TZ via integrative actions of auxin, NO and SLs.

  12. Nitric Oxide-Mediated Maize Root Apex Responses to Nitrate are Regulated by Auxin and Strigolactones.

    Science.gov (United States)

    Manoli, Alessandro; Trevisan, Sara; Voigt, Boris; Yokawa, Ken; Baluška, František; Quaggiotti, Silvia

    2015-01-01

    Nitrate (NO3 (-)) is a key element for crop production but its levels in agricultural soils are limited. Plants have developed mechanisms to cope with these NO3 (-) fluctuations based on sensing nitrate at the root apex. Particularly, the transition zone (TZ) of root apex has been suggested as a signaling-response zone. This study dissects cellular and molecular mechanisms underlying NO3 (-) resupply effects on primary root (PR) growth in maize, confirming nitric oxide (NO) as a putative modulator. Nitrate restoration induced PR elongation within the first 2 h, corresponding to a stimulation of cell elongation at the basal border of the TZ. Xyloglucans (XGs) immunolocalization together with Brefeldin A applications demonstrated that nitrate resupply induces XG accumulation. This effect was blocked by cPTIO (NO scavenger). Transcriptional analysis of ZmXET1 confirmed the stimulatory effect of nitrate on XGs accumulation in cells of the TZ. Immunolocalization analyses revealed a positive effect of nitrate resupply on auxin and PIN1 accumulation, but a transcriptional regulation of auxin biosynthesis/transport/signaling genes was excluded. Short-term nitrate treatment repressed the transcription of genes involved in strigolactones (SLs) biosynthesis and transport, mainly in the TZ. Enhancement of carotenoid cleavage dioxygenases (CCDs) transcription in presence of cPTIO indicated endogenous NO as a negative modulator of CCDs activity. Finally, treatment with the SLs-biosynthesis inhibitor (TIS108) restored the root growth in the nitrate-starved seedlings. Present report suggests that the NO-mediated root apex responses to nitrate are accomplished in cells of the TZ via integrative actions of auxin, NO and SLs.

  13. Nitric Oxide-Mediated Maize Root Apex Responses to Nitrate are Regulated by Auxin and Strigolactones

    Science.gov (United States)

    Manoli, Alessandro; Trevisan, Sara; Voigt, Boris; Yokawa, Ken; Baluška, František; Quaggiotti, Silvia

    2016-01-01

    Nitrate (NO3-) is a key element for crop production but its levels in agricultural soils are limited. Plants have developed mechanisms to cope with these NO3- fluctuations based on sensing nitrate at the root apex. Particularly, the transition zone (TZ) of root apex has been suggested as a signaling-response zone. This study dissects cellular and molecular mechanisms underlying NO3- resupply effects on primary root (PR) growth in maize, confirming nitric oxide (NO) as a putative modulator. Nitrate restoration induced PR elongation within the first 2 h, corresponding to a stimulation of cell elongation at the basal border of the TZ. Xyloglucans (XGs) immunolocalization together with Brefeldin A applications demonstrated that nitrate resupply induces XG accumulation. This effect was blocked by cPTIO (NO scavenger). Transcriptional analysis of ZmXET1 confirmed the stimulatory effect of nitrate on XGs accumulation in cells of the TZ. Immunolocalization analyses revealed a positive effect of nitrate resupply on auxin and PIN1 accumulation, but a transcriptional regulation of auxin biosynthesis/transport/signaling genes was excluded. Short-term nitrate treatment repressed the transcription of genes involved in strigolactones (SLs) biosynthesis and transport, mainly in the TZ. Enhancement of carotenoid cleavage dioxygenases (CCDs) transcription in presence of cPTIO indicated endogenous NO as a negative modulator of CCDs activity. Finally, treatment with the SLs-biosynthesis inhibitor (TIS108) restored the root growth in the nitrate-starved seedlings. Present report suggests that the NO-mediated root apex responses to nitrate are accomplished in cells of the TZ via integrative actions of auxin, NO and SLs. PMID:26834770

  14. Nitric Oxide-Mediated Maize Root Apex Responses to Nitrate are Regulated by Auxin and Strigolactones.

    Science.gov (United States)

    Manoli, Alessandro; Trevisan, Sara; Voigt, Boris; Yokawa, Ken; Baluška, František; Quaggiotti, Silvia

    2015-01-01

    Nitrate (NO3 (-)) is a key element for crop production but its levels in agricultural soils are limited. Plants have developed mechanisms to cope with these NO3 (-) fluctuations based on sensing nitrate at the root apex. Particularly, the transition zone (TZ) of root apex has been suggested as a signaling-response zone. This study dissects cellular and molecular mechanisms underlying NO3 (-) resupply effects on primary root (PR) growth in maize, confirming nitric oxide (NO) as a putative modulator. Nitrate restoration induced PR elongation within the first 2 h, corresponding to a stimulation of cell elongation at the basal border of the TZ. Xyloglucans (XGs) immunolocalization together with Brefeldin A applications demonstrated that nitrate resupply induces XG accumulation. This effect was blocked by cPTIO (NO scavenger). Transcriptional analysis of ZmXET1 confirmed the stimulatory effect of nitrate on XGs accumulation in cells of the TZ. Immunolocalization analyses revealed a positive effect of nitrate resupply on auxin and PIN1 accumulation, but a transcriptional regulation of auxin biosynthesis/transport/signaling genes was excluded. Short-term nitrate treatment repressed the transcription of genes involved in strigolactones (SLs) biosynthesis and transport, mainly in the TZ. Enhancement of carotenoid cleavage dioxygenases (CCDs) transcription in presence of cPTIO indicated endogenous NO as a negative modulator of CCDs activity. Finally, treatment with the SLs-biosynthesis inhibitor (TIS108) restored the root growth in the nitrate-starved seedlings. Present report suggests that the NO-mediated root apex responses to nitrate are accomplished in cells of the TZ via integrative actions of auxin, NO and SLs. PMID:26834770

  15. Detecting methane plumes with the APEX imaging spectrometer

    Science.gov (United States)

    Kuhlmann, Gerrit; Hueni, Andreas; Brunner, Dominik

    2016-04-01

    The Airborne Prism EXperiment (APEX) is an imaging spectrometer primarily designed for environmental remote sensing of the land surface but also allows observing atmospheric trace gases. To test if APEX can be used to detect strong methane plumes, the instrument was flown over a coal mining area near Ibbenbürren (Germany). According to the European Point source emission inventory (E-PRTR), emissions from the ventilation shafts of these coal mines are the largest single source of methane in Germany. We present the first measurements of methane ever taken by APEX making use of the absorption features of methane in the short-wave infrared spectral range. Our detection algorithm uses a matched filter to identify the presence of methane. The filter is tested in two spectral windows (1600-1700 nm and 2100-2500 nm) and for different spatial binning of pixels to improve the signal-to-noise. Using this approach, we could clearly identify two meandering methane plumes originating from the ventilation shafts of two coal mines (Bockradener Schacht and Theodor Schacht). The filter performed best for the spectral window from 1600-1700 nm with a binning of 10×10 pixels corresponding to a spatial resolution of about 35×25 m2 In conclusions, we could demonstrate that APEX is able to detect strong methane plumes. The results provide a basis for developing more sophisticated and quantitative methane retrievals.

  16. Primary mucocele of the petrous apex: MR appearance.

    Science.gov (United States)

    Larson, T L; Wong, M L

    1992-01-01

    Mucoceles of the petrous apex are rare. Their MR appearance varies depending on the degree of hydration or inspissation of the contents. Concise preoperative diagnosis is helpful, since mucoceles are better drained to the mastoids via an infralabyrinthine approach rather than the more risky middle cranial fossa approach used for cholesteatomas. PMID:1595444

  17. Ecosystem context and historical contingency in apex predator recoveries.

    Science.gov (United States)

    Stier, Adrian C; Samhouri, Jameal F; Novak, Mark; Marshall, Kristin N; Ward, Eric J; Holt, Robert D; Levin, Phillip S

    2016-05-01

    Habitat loss, overexploitation, and numerous other stressors have caused global declines in apex predators. This "trophic downgrading" has generated widespread concern because of the fundamental role that apex predators can play in ecosystem functioning, disease regulation, and biodiversity maintenance. In attempts to combat declines, managers have conducted reintroductions, imposed stricter harvest regulations, and implemented protected areas. We suggest that full recovery of viable apex predator populations is currently the exception rather than the rule. We argue that, in addition to well-known considerations, such as continued exploitation and slow life histories, there are several underappreciated factors that complicate predator recoveries. These factors include three challenges. First, a priori identification of the suite of trophic interactions, such as resource limitation and competition that will influence recovery can be difficult. Second, defining and accomplishing predator recovery in the context of a dynamic ecosystem requires an appreciation of the timing of recovery, which can determine the relative density of apex predators and other predators and therefore affect competitive outcomes. Third, successful recovery programs require designing adaptive sequences of management strategies that embrace key environmental and species interactions as they emerge. Consideration of recent research on food web modules, alternative stable states, and community assembly offer important insights for predator recovery efforts and restoration ecology more generally. Foremost among these is the importance of a social-ecological perspective in facilitating a long-lasting predator restoration while avoiding unintended consequences. PMID:27386535

  18. FoxO3a基因缺失致小鼠脾脏进行性慢性炎性反应的观察%Progressive chronic inflammatory reaction in spleens of FoxO3a gene knockout mice

    Institute of Scientific and Technical Information of China (English)

    邱红; 陈晨; 王若立; 罗红; 赵宝霞

    2012-01-01

    目的 观察FoxO3a基因缺失致小鼠脾脏进行性慢性炎性反应.方法 分别于16周、24周和12个月称取FoxO3a基因敲除小鼠和对照小鼠体重;于16、24和38周处死各组小鼠,取脾脏,肉眼观察脾脏变化,并制备脾细胞悬液,进行细胞计数,同时取各组小鼠卵巢、肺及皮下等组织,观察其炎性变化.流式细胞术检测24周的FoxO3a基因敲除小鼠和对照小鼠脾细胞中T、B淋巴细胞和Mac-1+细胞数量及百分率;Real-time PCR检测脾细胞中炎性细胞因子S100A8和S100A9基因mRNA的表达水平.结果 与对照小鼠相比,FoxO3a基因敲除小鼠脾脏明显肥大,且随年龄增加更加显著,卵巢、肺及皮下等组织内可见大量炎性细胞浸润,脾细胞数明显升高;基因敲除组老年小鼠的体重明显低于对照小鼠;FoxO3a基因敲除小鼠的脾细胞中T、B淋巴细胞数增加明显,但所占脾细胞的百分率无明显增加,Mac-1+细胞数和百分率均明显增加;S100A8和S100A9基因mRNA的表达水平明显高于对照小鼠.结论 FoxO3a基因缺失致小鼠脾脏进行性慢性炎性反应.%Objective To observe the progressive chronic inflammatory reaction induced by Fox03a gene knockout in mice. Methods The bodyweights of Fox03a gene knockout and normal control mice were weighed at ages of 16 weeks, 24 weeks and 12 months separately. Partial mice in the two groups were killed at ages of 16, 24 and 38 weeks respectively, of which the spleens were collected, observed visually, prepared into splenocytes and counted. Meanwhile, the inflammatory reactions in various tissues including ovary, lung and subcutaneous tissues were observed. The counts and percentages of T and B lymphocytes and Mac-1+ cells in spleens of FoxO3a gene knockout and control mice at age of 24 weeks were determined by flow cytometry, while the expression levels of S100A8 and S100A9 mRNAs in splenocytes by real-time PCR. Results Compared with those of control mice, the

  19. MES buffer affects Arabidopsis root apex zonation and root growth by suppressing superoxide generation in root apex

    Directory of Open Access Journals (Sweden)

    Tomoko eKagenishi

    2016-02-01

    Full Text Available In plants, growth of roots and root hairs is regulated by the fine cellular control of pH and reactive oxygen species. MES, 2-(N-morpholinoethanesulfonic acid as one of the Good’s buffers has broadly been used for buffering medium, and it is thought to suit for plant growth with the concentration at 0.1% (w/v because the buffer capacity of MES ranging pH 5.5-7.0 (for Arabidopsis, pH 5.8. However, many reports have shown that, in nature, roots require different pH values on the surface of specific root apex zones, namely meristem, transition zone and elongation zone. Despite the fact that roots always grow on a media containing buffer molecule, little is known about impact of MES on root growth. Here, we have checked the effects of different concentrations of MES buffer using growing roots of Arabidopsis thaliana. Our results show that 1% of MES significantly inhibited root growth, the number of root hairs and length of meristem, whereas 0.1% promoted root growth and root apex area (region spanning from the root tip up to the transition zone. Furthermore, superoxide generation in root apex disappeared at 1% of MES. These results suggest that MES disturbs normal root morphogenesis by changing the reactive oxygen species (ROS homeostasis in root apex.

  20. MES Buffer Affects Arabidopsis Root Apex Zonation and Root Growth by Suppressing Superoxide Generation in Root Apex.

    Science.gov (United States)

    Kagenishi, Tomoko; Yokawa, Ken; Baluška, František

    2016-01-01

    In plants, growth of roots and root hairs is regulated by the fine cellular control of pH and reactive oxygen species (ROS). MES, 2-(N-morpholino)ethanesulfonic acid as one of the Good's buffers has broadly been used for buffering medium, and it is thought to suit for plant growth with the concentration at 0.1% (w/v) because the buffer capacity of MES ranging pH 5.5-7.0 (for Arabidopsis, pH 5.8). However, many reports have shown that, in nature, roots require different pH values on the surface of specific root apex zones, namely meristem, transition zone, and elongation zone. Despite the fact that roots always grow on a media containing buffer molecule, little is known about impact of MES on root growth. Here, we have checked the effects of different concentrations of MES buffer using growing roots of Arabidopsis thaliana. Our results show that 1% of MES significantly inhibited root growth, the number of root hairs and length of meristem, whereas 0.1% promoted root growth and root apex area (region spanning from the root tip up to the transition zone). Furthermore, superoxide generation in root apex disappeared at 1% of MES. These results suggest that MES disturbs normal root morphogenesis by changing the ROS homeostasis in root apex. PMID:26925066

  1. Knockout Reaction Mechanism for 6He+%Knockout Reaction Mechanism for 6He+

    Institute of Scientific and Technical Information of China (English)

    吕林辉; 叶沿林; 曹中鑫; 肖军; 江栋兴; 郑涛; 华辉; 李智焕; 葛俞成; 李湘庆; 楼建玲; 李阔昂; 李奇特; 乔锐; 游海波; 陈瑞九

    2012-01-01

    A knockout reaction experiment was carried out by using the 6He beam at 82.5 MeV/nucleon impinging on CH2 and C targets. The a core fragments at forward angles were detected in coincidence with the recoiled protons at larger angles. From this exclusive measure- ment the valence nucleon knockout mechanism and the core knockout mechanism are separated. This study provides a basis for the exclusive spectroscopic investigation of the exotic nuclei.

  2. Reproduction and genotype identification of corticotropin-releasing hormone gene knockout mice%促肾上腺皮质激素释放激素基因敲除小鼠的繁殖与基因型鉴定

    Institute of Scientific and Technical Information of China (English)

    王海燕; 刘庆; 钟河江; 杨策; 黄苏娜; 严军; 蒋建新

    2011-01-01

    目的 探讨促肾上腺皮质激素释放激素(CRH)基因敲除(KO)小鼠饲养、繁殖及基因型鉴定的方法.方法 从美国Jackson实验室引进CRH KO小鼠,按照遗传学规则,对杂合子型(CRH+/-)小鼠进行配对繁殖,提取幼鼠尾部组织全基因组DNA,通过聚合酶链反应(PCR)对幼鼠基因型进行鉴定.结果 CRH KO纯合子型(CRH-/-)小鼠的繁殖和饲养均获得成功,采用PCR成功地对所获得的小鼠进行基因分析,在子代小鼠中存在野生纯合子型(CRH+/+)、杂合子型(CRH+/-)及CRH KO纯合子型(CRH-/-)小鼠.CRH-/-小鼠较另外2种基因型小鼠存活率明显下降,但3种基因型小鼠在出生后10 d及30 d体质量无明显差异.结论 正确的饲养繁殖以及鉴定方法可从杂合子型(CRH+/-)小鼠中获得CRH KO纯合子型(CRH-/-)小鼠.%Objective To explore the methods of breeding, reproductin and genotype identification of corticotropin-releasing hormone ( CRH)knockout( KO) mice.Methods CRH knockout mice were obtained from Jackson laboratory in USA.Heterozygous type (CRH+/- )mice were inbreeded according to genetic rules to yield CRH knockout mice.The genotypes of offspring were identified by polymerase chain reaction(PCR)using genomic DNA extracted from tissue of mice tails.Results Both breeding and reproductin of CRH KO heterozygous type(CRH+/- )mice were successful.PCR was used successfully for genetic analysis in mice obtained.There were wild homozygous genotype( CRH+ /+ ) , heterozygous genotype ( CRH + /- ) and CRH KO homozygous genotype( CRH-/- )in the offspring.Compared with other two genotype mice,survival rate of CRH- /- mice were significantly decreased.however, body mass of the three genotypes mice had no significant difference at 10 and 30 days after birth.Conclusion Appropriate reproductin , breeding and identification are effective methods to obtain CRH KO homozygous genotype( CRH -/- ) mice from heterozygous genotype( CRH+ / - ) mice.

  3. Apolipoprotein E knockout as the basis for mouse models of dyslipidemia-induced neuropathy.

    Science.gov (United States)

    Hinder, Lucy M; Vincent, Andrea M; Hayes, John M; McLean, Lisa L; Feldman, Eva L

    2013-01-01

    Dyslipidemia has been identified as an important pathogenic risk factor for diabetic neuropathy, but current animal models do not adequately reproduce the lipid profile observed in human diabetics (increased triglycerides with an elevated LDL-cholesterol and reduced HDL-cholesterol). High fat feeding of mice produces hyperlipidemia, but mice are resistant to increases in the LDL to HDL ratio, reducing the potential for peripheral lipid deposits to impact neuropathy, as is postulated to occur in human subjects. Genetic manipulations provide an alternative approach to reproducing a neuropathic plasma lipid profile. Based on findings from the atherosclerosis literature, we began with knockout of ApoE. Since knockout of ApoE alone only partially mimics the human diabetic lipid profile, we examined the impact of its combination with a well-characterized model of type 2 diabetes exhibiting neuropathy, the db/db mouse. We added further gene manipulations to increase hyperlipidemia by using mice with both ApoE and ApoB48 knockout on the ob/+ (leptin mutation) mice. In all of these models, we found that either the db/db or ob/ob genotypes had increased body weight, hyperlipidemia, hyperglycemia, and evidence of neuropathy compared with the control groups (db/+ or ob/+, respectively). We found that ApoE knockout combined with leptin receptor knockout produced a lipid profile most closely modeling human dyslipidemia that promotes neuropathy. ApoE knockout combined with additional ApoB48 and leptin knockout produced similar changes of smaller magnitude, but, notably, an increase in HDL-cholesterol. Our data suggest that the overall effects of ApoE knockout, either directly upon nerve structure and function or indirectly on lipid metabolism, are insufficient to significantly alter the course of diabetic neuropathy. Although these models ultimately do not deliver optimal lipid profiles for translational diabetic neuropathy research, they do present glycemic and lipid profile

  4. G 蛋白偶联受体56基因敲除抑制少突胶质前体细胞成熟%Knockout of GPR56 gene inhibits maturation of oligodendrocyte progeni-tor cells

    Institute of Scientific and Technical Information of China (English)

    邓医宇; 朱高峰; 方明; 曾文新; 蒋文新; 曾红科

    2014-01-01

    AIM:To explore the effect of G-protein-coupled protein 56 (GPR56) gene knockout on axonal my-elination and the maturation of oligodendrocyte progenitor cells (OPCs) in the corpus callosum of mouse brain.METH-ODS:Thirty-six GPR56+/-and GPR56-/-mice were selected and divided into GPR56+/-group and GPR56-/-group (18 mice in each group).According to the postnatal days , the mice in each group were further divided into P7, P14, P21 and P28 subgroups.Myelin formation in the corpus callosum of P14, P21 and P28 GPR56+/-and GPR56-/-mice was ob-served by FluoroMyelin staining .The number of myelinated axons and thickness of myelin sheaths were measured by elec -tron microscopy.The numbers of platelet-derived growth factor alpha receptor-positive (PDGF-aR+) and proteolipid pro-tein-positive (PLP+) cells in the corpus callosum of GPR56+/-and GPR56-/-mice were compared by the methods of im-munofluorescence and in situ hybridization.GPR56+/- and GPR56-/- OPCs were cultured using P1 GPR56+/- and GPR56 -/-mouse brain cortex and induced differentiation and maturation in vitro.The percentage of GPR56+/- and GPR56 -/-O4+cells in pro-oligodendroblast, immature oligodendrocyte and mature oligodendrocyte stages was compared by O4 immunostaining.RESULTS:The myelin formation was obviously reduced in the corpus callosum of P 14, P21 and P28 GPR56-/-mice as compared with GPR56+/-mice.The number of myelinated axons was obviously reduced and the g-ratio value was increased significantly in the corpus callosum of P 28 GPR56-/-mice.No significant difference of the PDGF-aR+cell number in the corpus callosum between P 7 d GPR56+/-and GPR56-/-mouse brains was observed.The number of PLP+ cells was significantly decreased in the corpus callosum of P 28 GPR56-/- mice as compared with GPR56 +/-mice.The percentage of GPR56-/-O4+cells in pro-oligodendroblast stages was obviously higher than that of GPR56 +/-O4 +cells.On the contrary, the percentages of GPR56 -/-O4 +cells in immature oligodendrocyte and

  5. Wholly Endoscopic Permeatal Removal of a Petrous Apex Cholesteatoma

    Directory of Open Access Journals (Sweden)

    Todd Kanzara

    2014-01-01

    Full Text Available We report a case of a petrous apex cholesteatoma which was managed with a wholly endoscopic permeatal approach. A 63-year-old Caucasian male presented with a 10-year history of right-sided facial palsy and profound deafness. On examination in our clinic, the patient had a grade VI House-Brackmann paresis, otoscopic evidence of attic cholesteatoma behind an intact drum, and extensive scarring of the face from previous facial reanimation surgery. Imaging review was suggestive of petrous apex cholesteatoma. An initial decision to manage the patient conservatively was later reviewed on account of the patient suffering recurrent epileptic seizures. A wholly endoscopic permeatal approach was used with successful outcomes. In addition to the case report we also provide a brief description of the technique and a review of the relevant literature.

  6. APEX 1 mm line survey of the Orion Bar

    CERN Document Server

    Leurini, S; Thorwirth, S; Parise, B; Schilke, P; Comito, C; Wyrowski, F; Güsten, R; Bergman, P; Menten, K M; Nyman, L A A

    2006-01-01

    Unbiased molecular line surveys are a powerful tool for analyzing the physical and chemical parameters of astronomical objects and are the only means for obtaining a complete view of the molecular inventory for a given source. The present work stands for the first such investigation of a photon-dominated region. The first results of an ongoing millimeter-wave survey obtained towards the Orion Bar are reported. The APEX telescope in combination with the APEX-2A facility receiver was employed in this investigation. We derived the physical parameters of the gas through LVG analyses of the methanol and formaldehyde data. Information on the sulfur and deuterium chemistry of photon-dominated regions is obtained from detections of several sulfur-bearing molecules and DCN.

  7. Selective Deactivation of Gibberellins below the Shoot Apex is Critical to Flowering but Not to Stem Elongation of Lolium

    Institute of Scientific and Technical Information of China (English)

    Rod W.King; Lewis N.Mander; Torben Asp; Colleen P. MacMillan; Cheryl A.Blundell; Lloyd T.Evans

    2008-01-01

    Gibberellins (GAs) cause dramatic increases in plant height and a genetic block in the synthesis of GA1 explains the dwarfing of Mendel's pea.For flowering,it is GAs which is important in the long-day (LD) responsive grass,Lolium.As we show here,GA1 and GA4 are restricted in their effectiveness for flowering because they are deactivated by C-2 hydroxylation below the shoot apex.In contrast,GAs is effective because of its structural protection at C-2.Excised vegetative shoot tips rapidly degrade [14C]GA1,[14C]GA4,and [14C]GA20 (>80% in 6 h),but not [14C]GA5.Coincidentally,genes encoding two 2β-oxidases and a putative 16-17-epoxidase were most expressed just below the shoot apex (4 mm),expression of these GA deactivation genes is reduced,so allowing GA1 and GA4 to promote sub-apical stem elongation.Subsequently,GA degradation declines in florally induced shoot tips and these GAs can become active for floral development.Structural changes which stabilize GA4 confirm the link between florigenicity and restricted GA 2β-hydroxylation (e.g.2α-hydroxylation and C-2 di-methylation).Additionally,a 2-oxidase inhibitor (Trinexapac Ethyl) enhanced the activity of applied GA4,as did limiting C-16,17 epoxidation in 16,17-dihydro GAs or after C-13 hydroxylation.Overall,deactivation of GA1 and GA4 just below the shoot apex effectively restricts their florigenicity in Lolium and,conversely,with GAs,C-2 and C-13 protection against deactivation allows its high florigenicity.Speculatively,such differences in GA access to the shoot apex of grasses may be important for separating floral induction from inflorescence emergence and thus could influence their survival under conditions of herbivore predation.

  8. Mucormycosis with Orbital Apex Syndrome in a Renal Transplant Recipient

    Directory of Open Access Journals (Sweden)

    Ebru Kursun

    2015-06-01

    Full Text Available Mucormycosis is a rarely encountered invasive fungal infection with high mortality.Solid organ transplantation is one of the risk factors for mucormycosis. Mucormycosis can be classified in six different groups according to the anatomical localization; rhinocerebral, pulmonary, cutaneous, gastrointestinal, disseminated, and other less common involvements. This paper presented a mucormycosis case with rhinoorbitocerebral involvementin a renal transplantation receiver, which manifested with orbital apex syndrome. [Cukurova Med J 2015; 40(2.000: 384-389

  9. Analysis of Apex Seal Friction Power Loss in Rotary Engines

    Science.gov (United States)

    Handschuh, Robert F.; Owen, A. Karl

    2010-01-01

    An analysis of the frictional losses from the apex seals in a rotary engine was developed. The modeling was initiated with a kinematic analysis of the rotary engine. Next a modern internal combustion engine analysis code was altered for use in a rotary engine to allow the calculation of the internal combustion pressure as a function of rotor rotation. Finally the forces from the spring, inertial, and combustion pressure on the seal were combined to provide the frictional horsepower assessment.

  10. Crossing Latitudes—Long-Distance Tracking of an Apex Predator

    OpenAIRE

    Ferreira, Luciana C.; Michele Thums; Jessica J Meeuwig; Vianna, Gabriel M. S.; John Stevens; Rory McAuley; Meekan, Mark G

    2015-01-01

    Tiger sharks (Galeocerdo cuvier) are apex predators occurring in most tropical and warm temperate marine ecosystems, but we know relatively little of their patterns of residency and movement over large spatial and temporal scales. We deployed satellite tags on eleven tiger sharks off the north-western coast of Western Australia and used the Brownian Bridge kernel method to calculate home ranges and analyse movement behaviour. One individual recorded one of the largest geographical ranges of m...

  11. APEX-SZ first light and instrument status

    Science.gov (United States)

    Dobbs, M.; Halverson, N. W.; Ade, P. A. R.; Basu, K.; Beelen, A.; Bertoldi, F.; Cohalan, C.; Cho, H. M.; Güsten, R.; Holzapfel, W. L.; Kermish, Z.; Kneissl, R.; Kovács, A.; Kreysa, E.; Lanting, T. M.; Lee, A. T.; Lueker, M.; Mehl, J.; Menten, K. M.; Muders, D.; Nord, M.; Plagge, T.; Richards, P. L.; Schilke, P.; Schwan, D.; Spieler, H.; Weiss, A.; White, M.

    2006-12-01

    The APEX-SZ instrument is designed for the discovery and study of galaxy clusters at mm-wavelengths using the Sunyaev Zel'dovich effect. The receiver consists of 320 superconducting transition edge sensor (TES) bolometers cooled to 250 mK with the combination of a three stage He sorption fridge and mechanical pulse tube cooler. The detectors are instrumented with a frequency domain multiplexing readout system. The receiver is mounted on the 12 m APEX telescope located at 5100 m on the Atacama plateau in Chile. For the first light engineering deployment of December 2005, the receiver was configured with a 55 element wedge of the bolometers and operating in the 150 GHz atmospheric window. During the engineering run we achieved significant milestones in our instrumentation development efforts, including celestial observations with a monolithically fabricated TES bolometer array cooled with a mechanical cooler and successful implementation of a SQUID-based MHz AC-biased readout. These technology demonstrations point the way toward future large TES bolometer array instruments. Here we describe the results of this deployment and future plans for the APEX-SZ instrument.

  12. Hearing Outcomes after Surgical Drainage of Petrous Apex Cholesterol Granuloma.

    Science.gov (United States)

    Rihani, Jordan; Kutz, J Walter; Isaacson, Brandon

    2015-06-01

    Objective This study aims to assess the hearing outcomes of patients undergoing surgical management of petrous apex cholesterol granuloma and to discuss the role of otic capsule-sparing approaches in drainage of petrous apex cholesterol granulomas. Design Retrospective case series. Setting Tertiary care medical center. Participants Eight patients underwent surgery for presumed or definitive cholesterol granuloma between 2002 and 2011 and met the inclusion criteria for this study. Main Outcome Measures Pre- and postoperative audiogram results as measured by pure tone thresholds and word recognition scores. Results Four patients (50%) demonstrated improvement in speech discrimination. One patient had an increase from 0 to 67% in word recognition scores. Four patients (50%) demonstrated worsening of pure tone thresholds, including two patients with anacusis. Conclusion Perilabyrinthine drainage of petrous apex cholesterol granulomas may result in hearing preservation or hearing improvement, even in the setting of otic capsule erosion. Patients should be counseled about the potential risk of significant hearing loss. PMID:26225297

  13. Fmr1基因敲除小鼠耳蜗的GABAα1受体的表达%Expression of GABAα1 receptor of cochlea in FMR1 gene knock-out mice

    Institute of Scientific and Technical Information of China (English)

    李敏雄; 杜娜; 孙卫文; 黄月玲; 沈岩松; 戴丽军; 陈盛强; 马钊恩; 张建国

    2012-01-01

    Objective To observe cochlea morphology and expression of GABA a 1 receptor of cochlea in 4 weeks FMR1 KO mice and WT mice. Methods Four-week old Fmrl knockout mice were identified using the PCR technique.and immunohistochemistry to compare with the changes of expression of GABA a 1 receptor between FMR1 KO mice and WT mice cochlea. Results There were no difference in cochlea morphology between FMR1 KO mice and WT mice by HE dyeing. The expression of GABA a 1 receptor in cochlear in FMR-1K0 mice was decreased. Conclusion The expression of GABA a 1 receptor is incerased in cochlear in four-week old FMR-1K0 mice that might be associated with audiogenic seizure susceptibility of Fmrl knockout mice.%目的 对4周龄Fmr1基因敲除小鼠耳蜗的GABAα1受体表达进行观察,探讨耳蜗GABAα1受体的表达是否受FMRP的影响.方法 使用PCR技术对Fmr1基因敲除小鼠鉴定后,对4周龄的Fmr1基因敲除小鼠和野生型小鼠进行耳蜗的GABAα1受体免疫组织化学的表达观察,数据采用多因素方差分析处理.结果 耳蜗HE染色结果:4周龄组KO鼠较WT鼠形态学观察无差异.4周龄KO小鼠的耳蜗中GABAα1受体表达的平均阳性细胞数均低于WT小鼠,P<0.01,差异具有统计学意义.结论 GABAα1受体表达的降低可能与FMR1基因KO小鼠听源性惊厥发病有关.

  14. Brief Report: Altered Social Behavior in Isolation-Reared "Fmr1" Knockout Mice

    Science.gov (United States)

    Heitzer, Andrew M.; Roth, Alexandra K.; Nawrocki, Lauren; Wrenn, Craige C.; Valdovinos, Maria G.

    2013-01-01

    Social behavior abnormalities in Fragile X syndrome (FXS) are characterized by social withdrawal, anxiety, and deficits in social cognition. To assess these deficits, a model of FXS, the "Fmr1" knockout mouse ("Fmr1" KO), has been utilized. This mouse model has a null mutation in the fragile X mental retardation 1 gene ("Fmr1") and displays…

  15. Activation of NADPH-recycling systems in leaves and roots of Arabidopsis thaliana under arsenic-induced stress conditions is accelerated by knock-out of Nudix hydrolase 19 (AtNUDX19) gene.

    Science.gov (United States)

    Corpas, Francisco J; Aguayo-Trinidad, Simeón; Ogawa, Takahisa; Yoshimura, Kazuya; Shigeoka, Shigeru

    2016-03-15

    NADPH is an important cofactor in cell growth, proliferation and detoxification. Arabidopsis thaliana Nudix hydrolase 19 (AtNUDX19) belongs to a family of proteins defined by the conserved amino-acid sequence GX5-EX7REUXEEXGU which has the capacity to hydrolyze NADPH as a physiological substrate in vivo. Given the importance of NADPH in the cellular redox homeostasis of plants, the present study compares the responses of the main NADPH-recycling systems including NADP-isocitrate dehydrogenase (ICDH), glucose-6-phosphate dehydrogenase (G6PDH), 6-phosphogluconate dehydrogenase (6PGDH) and NADP-malic enzyme (ME) in the leaves and roots of Arabidopsis wild-type (Wt) and knock-out (KO) AtNUDX19 mutant (Atnudx19) plants under physiological and arsenic-induced stress conditions. Two major features were observed in the behavior of the main NADPH-recycling systems: (i) under optimal conditions in both organs, the levels of these activities were higher in nudx19 mutants than in Wt plants; and, (ii) under 500μM AsV conditions, these activities increase, especially in nudx19 mutant plants. Moreover, G6PDH activity in roots was the most affected enzyme in both Wt and nudx19 mutant plants, with a 4.6-fold and 5.0-fold increase, respectively. In summary, the data reveals a connection between the absence of chloroplastic AtNUDX19 and the rise in all NADP-dehydrogenase activities under physiological and arsenic-induced stress conditions, particularly in roots. This suggests that AtNUDX19 could be a key factor in modulating the NADPH pool in plants and consequently in redox homeostasis. PMID:26878367

  16. Bone phenotypes of P2 receptor knockout mice

    DEFF Research Database (Denmark)

    Orriss, Isabel; Syberg, Susanne; Wang, Ning;

    2011-01-01

    The action of extracellular nucleotides is mediated by ionotropic P2X receptors and G-protein coupled P2Y receptors. The human genome contains 7 P2X and 8 P2Y receptor genes. Knockout mice strains are available for most of them. As their phenotypic analysis is progressing, bone abnormalities have...... been observed in an impressive number of these mice: distinct abnormalities in P2X7-/- mice, depending on the gene targeting construct and the genetic background, decreased bone mass in P2Y1-/- mice, increased bone mass in P2Y2-/- mice, decreased bone resorption in P2Y6-/- mice, decreased bone...... formation and bone resorption in P2Y13-/- mice. These findings demonstrate the unexpected importance of extracellular nucleotide signalling in the regulation of bone metabolism via multiple P2 receptors and distinct mechanisms involving both osteoblasts and osteoclasts....

  17. Vectors Building and Usage for Gene Knockout, Protein Expression and Fluorescent Fusion Protein in The Rice Blast Fungus%适用于稻瘟病菌基因敲除、过表达和荧光融合蛋白表达载体的构建和使用

    Institute of Scientific and Technical Information of China (English)

    李海娇; 卢建平; 刘小红; 张莉林; 林福呈

    2012-01-01

    This study is to supply a series of vectors for gene knockout, overexpression, and expressing fluorescent fusion proteins in the rice blast fungus. These vectors should be easily worked, time-saving, reliable, and conveniently for the research in the gene function of Magnaporthe oryzae. PCR, enzyme digestion, ligation a^id transformation were used to construct the plasmids. We cloned two promoters (SOD1 promoter and H3 promoter) which strongly expressed in the mycelia and conidia, built 3 vectors for knockout (pBS-SUR, pBS-BAS,pBS-NEO), 4 vectors for overexpression (pKD5, pKD6, pKD61 and pKD8), and 7 vectors for fluorescent fusion protein expression (pKD5-GFP, pKD5-RED, pKD6-GFP, pKD6-RED, pKD7-RED, pKD8-GFP and pKD8-RED) in M. Oryzae and other filamentous fungi. These plasmids could be introduced into the fungi via protoplast transformation, or A grobacterium tumefaciens -mediated transformation. The knockout mutants could be identified by PCR, Southern blot and Western blot, the fluorescence of the transformants was observed under the fluorescent microscope, and the expression level of genes in the transformants was assayed by Real-time PCR. We have used knockout constructs built by these knockout vectors (pBS-SUR, pBS-BAS, pBS-NEO) and pBS-HPHl together to knockout 4 genes simultaneously in M. Oryzae. After transforming pKD5-RED, pKD6-GFP or pKD6-RED into M. Oryzae via A grobacterium tumefac iens -mediated transformation, green or red fluorescent fusion proteins under the control of SOD1 or H3 promoter were expressed strongly in the hyphae; Real-time PCR results showed eGFP mRNA or DsRED2 mRNA level promoted by SOD1 promoter was 2.5 or 5.4 folds of (I-tubulin, and DsRED2 mRNA level promoted by H3 promoter was 20.8 folds of fi-tubulin in the mycelia. MoATG8-DsRED2 fusion protein produced by pKD6-RED could locate M0ATG8 exactly in the nearby of vacuoles. Observation on DsRED2 fluorescent protein under micrpscope showed that DsRED2 under control of SOD1 promoter

  18. Robust and sensitive analysis of mouse knockout phenotypes.

    Directory of Open Access Journals (Sweden)

    Natasha A Karp

    Full Text Available A significant challenge of in-vivo studies is the identification of phenotypes with a method that is robust and reliable. The challenge arises from practical issues that lead to experimental designs which are not ideal. Breeding issues, particularly in the presence of fertility or fecundity problems, frequently lead to data being collected in multiple batches. This problem is acute in high throughput phenotyping programs. In addition, in a high throughput environment operational issues lead to controls not being measured on the same day as knockouts. We highlight how application of traditional methods, such as a Student's t-Test or a 2-way ANOVA, in these situations give flawed results and should not be used. We explore the use of mixed models using worked examples from Sanger Mouse Genome Project focusing on Dual-Energy X-Ray Absorptiometry data for the analysis of mouse knockout data and compare to a reference range approach. We show that mixed model analysis is more sensitive and less prone to artefacts allowing the discovery of subtle quantitative phenotypes essential for correlating a gene's function to human disease. We demonstrate how a mixed model approach has the additional advantage of being able to include covariates, such as body weight, to separate effect of genotype from these covariates. This is a particular issue in knockout studies, where body weight is a common phenotype and will enhance the precision of assigning phenotypes and the subsequent selection of lines for secondary phenotyping. The use of mixed models with in-vivo studies has value not only in improving the quality and sensitivity of the data analysis but also ethically as a method suitable for small batches which reduces the breeding burden of a colony. This will reduce the use of animals, increase throughput, and decrease cost whilst improving the quality and depth of knowledge gained.

  19. Petrous apex cephalocele and empty sella: Is there any relation?

    Energy Technology Data Exchange (ETDEWEB)

    Alorainy, Ibrahim A. [Department of Radiology and Diagnostic Imaging, College of Medicine and King Khalid University Hospital, King Saud University, PO Box: 9047, Riyadh 11413 (Saudi Arabia)]. E-mail: alorainy@ksu.edu.sa

    2007-06-15

    Objective: To document the presence of incidental petrous apex cephalocele (PAC) in association with empty sella in a group of patients and propose an etiologic/pathologic relation between the two lesions. Materials and methods: Retrospective review of our imaging archive for the period from October 2001 to October 2006 revealed five patients with PAC (four females and one male; age range, 25-60 years; mean, 47 years). All patients underwent enhanced MR examination of the skull base and four of them underwent CT examination. Lesions were evaluated for size, content, signal intensity, enhancement, and relation to Meckel's cave and petrous apex. Images were also evaluated for the presence of empty sella. Results: The presenting symptoms in all patients were not attributable to PAC. None of the patients had symptoms related to the trigeminal nerve or history of CSF leak. Four patients had bilateral PAC and one had left PAC (total nine lesions). The lesions ranged from 6 mm to 15 mm (mean 9 mm) in the maximum diameter. All lesions were centered posterolateral to Meckel's cave and had low attenuation on CT with sharply demarcated margins. No lesion reached the inner ear structures, internal auditory canal, or mastoid air cells. On MR imaging, all lesions demonstrated CSF signal intensity that is continuous with that of the Meckel's cave. Only the periphery of the lesions demonstrated mild enhancement. All patients had empty sella. One patient had small arachnoid cysts in the middle cranial fossa, bilaterally. Conclusion: PAC and empty sella are similar mechanically in terms of CSF extension and erosion into petrous apex and sella, respectively. Both conditions are seen predominantly in females and have been reported in association with CSF leak, which raises a possibility of etiologic/pathologic relation between the two.

  20. Knockout of vlf-1 Gene in Bombyx mori Nucleopolyhedrovirus Using Red Recombination System and Its Influence on Viral Replication%利用Red重组系统敲除家蚕核型多角体病毒的vlf-1基因及对病毒复制的影响

    Institute of Scientific and Technical Information of China (English)

    杜超逸; 于威; 全滟平; 陈健; 聂作明; 吕正兵; 张耀洲

    2013-01-01

    In order to explore the function of BmNPV (Bombyx mori nucleopolyhedrovirus) vlf-1 gene in viral replication, Red recombinant system and BmNPV Bacmid were utilized to have a quick knockout of BmNPV vlf-1 gene in E. coli DHIOBac. Firstly, pKD46 plasmid capable of expressing Red recombinase was isolated from E. coli BW25113 and transformed into E. coli DH10Bac to obtain E. coli DH10Bac (containing pKD46), which could be used for targeting BmNPV gene. Then, for homologous recombination of the target gene, a pair of primers with 60 bp in length was designed, with 40 bp at their 5' end homologous to the left and right arm of vlf-1 gene respectively, and 20 bp at their 3' end homologous to the beginning and ending sequence of cat gene respectively. A linear fragment was amplified by PCR from plasmid pKD3 (containing cat gene) using these primers to obtain the target linear fragment with homologous arms to vlf-1. The linear fragment was transformed into DH10Bac (pKD46) and homologous recombination occurred between the linear fragment and vlf-1 gene in Bacmid DNA with the help of Red recombinase. Two specific PCR primer pairs were used to verify the successful replacement of vlf-1 by cat gene. Then the vlf-1 knockout Bacmid DNA was transfected into BmN cells, and qPCR was employed to analyze the influence of vlf-1 knockout on viral replication. The result indicated that vlf-1 knockout had no influence on initiation of BmNPV DNA replication, but might affect subsequent virus assembly and infection.%利用Red重组系统和家蚕核型多角体病毒(BmNPV) Bacmid在E.coli DH10Bac中快速敲除BmNPV极晚期表达因子基因vlf-1,调查对病毒复制的影响,以探究BmNPV vlf-1基因的功能.首先从E.coli BW25113中提取能表达Red重组酶的质粒pKD46,将其转化到E.coli DH10Bac中,获得可用于BmNPV基因打靶的DH10Bac(含有质粒pKD46)菌株;再为发生基因同源重组设计一对长60 bp的引物,5 '端长40 bp,分别为vlf-1

  1. APEX sub-mm monitoring of gamma-ray blazars

    CERN Document Server

    Larsson, S; Weiss, A; Angelakis, E; Krichbaum, T P; Nestoras, I; Zensus, J A; Axelsson, M; Nilsson, D; Ryde, F; Hjalmarsdotter, L; Larsson, J; Lundgren, A; Mac-Auliffe, F; Parra, R; Siringo, G

    2012-01-01

    So far, no systematic long-term blazar monitoring programs and detailed variability studies exist at sub-mm wavelengths. Here, we present a new sub-mm blazar monitoring program using the APEX 12-m telescope. A sample of about 40 gamma-ray blazars has been monitored since 2007/2008 with the LABOCA bolometer camera at 345 GHz. First light curves, preliminary variability results and a first comparison with the longer cm/mm bands (F-GAMMA program) are presented, demonstrating the extreme variability characteristics of blazars at such short wavelengths.

  2. Apex-angle-dependent resonances in triangular split ring resonators

    CERN Document Server

    Burnett, Max A

    2016-01-01

    Along with other frequency selective structures [1] (circles and squares), triangular split-ring resonators (TSRRs) only allow frequencies near the center resonant frequency to propagate. Further, TSRRs are attractive due to their small surface area [2], comparatively, and large quality factors (Q) factors as previously investigated by Gay-Balmaz, et al. [3]. In this work we examine the effects of varying the apex angle on the resonant frequency, the Q factor, and the phase shift imparted by the TSRR element within the GHz frequency regime.

  3. TFF3 knockout in human pituitary adenoma cell HP75 facilitates cell apoptosis via mitochondrial pathway.

    Science.gov (United States)

    Gao, Feng; Pan, Suxia; Liu, Bing; Zhang, Huanzhi

    2015-01-01

    Trefoil factor 3 (TFF3), a regulatory protein composed of 59 amino acids, has been suggested to be involved in pathogenesis, proliferation, differentiation, invasion, migration and apoptosis in multiple malignant tumors. This study thus investigated the effect of TFF3 knockout in human pituitary adenoma cell line HP75 on cell apoptosis and related pathways. RNA interference approach was used to knock down the expression of TFF3 protein. The gene silencing was validated by RNA denaturing gel electrophoresis and Western blotting. The effect of TFF3 knockout on cell apoptosis was analyzed by Western blotting and flow cytometry. TFF3 protein level in pituitary adenoma was about 3.61 ± 0.48 folds of that in normal tissues (P TFF3, the apoptotic ration was significantly elevated (P TFF3 protein knockout can facilitate apoptosis of human pituitary adenoma HP75 cells via mitochondrial pathway.

  4. TFF3 knockout in human pituitary adenoma cell HP75 facilitates cell apoptosis via mitochondrial pathway

    Science.gov (United States)

    Gao, Feng; Pan, Suxia; Liu, Bing; Zhang, Huanzhi

    2015-01-01

    Trefoil factor 3 (TFF3), a regulatory protein composed of 59 amino acids, has been suggested to be involved in pathogenesis, proliferation, differentiation, invasion, migration and apoptosis in multiple malignant tumors. This study thus investigated the effect of TFF3 knockout in human pituitary adenoma cell line HP75 on cell apoptosis and related pathways. RNA interference approach was used to knock down the expression of TFF3 protein. The gene silencing was validated by RNA denaturing gel electrophoresis and Western blotting. The effect of TFF3 knockout on cell apoptosis was analyzed by Western blotting and flow cytometry. TFF3 protein level in pituitary adenoma was about 3.61 ± 0.48 folds of that in normal tissues (P TFF3, the apoptotic ration was significantly elevated (P TFF3 protein knockout can facilitate apoptosis of human pituitary adenoma HP75 cells via mitochondrial pathway. PMID:26823779

  5. Microarray analysis of E9.5 reduced folate carrier (RFC1; Slc19a1 knockout embryos reveals altered expression of genes in the cubilin-megalin multiligand endocytic receptor complex

    Directory of Open Access Journals (Sweden)

    Bauer Linda K

    2008-04-01

    Full Text Available Abstract Background The reduced folate carrier (RFC1 is an integral membrane protein and facilitative anion exchanger that mediates delivery of 5-methyltetrahydrofolate into mammalian cells. Adequate maternal-fetal transport of folate is necessary for normal embryogenesis. Targeted inactivation of the murine RFC1 gene results in post-implantation embryolethality, but daily folic acid supplementation of pregnant dams prolongs survival of homozygous embryos until mid-gestation. At E10.5 RFC1-/- embryos are developmentally delayed relative to wildtype littermates, have multiple malformations, including neural tube defects, and die due to failure of chorioallantoic fusion. The mesoderm is sparse and disorganized, and there is a marked absence of erythrocytes in yolk sac blood islands. The identification of alterations in gene expression and signaling pathways involved in the observed dysmorphology following inactivation of RFC1-mediated folate transport are the focus of this investigation. Results Affymetrix microarray analysis of the relative gene expression profiles in whole E9.5 RFC1-/- vs. RFC1+/+ embryos identified 200 known genes that were differentially expressed. Major ontology groups included transcription factors (13.04%, and genes involved in transport functions (ion, lipid, carbohydrate (11.37%. Genes that code for receptors, ligands and interacting proteins in the cubilin-megalin multiligand endocytic receptor complex accounted for 9.36% of the total, followed closely by several genes involved in hematopoiesis (8.03%. The most highly significant gene network identified by Ingenuity™ Pathway analysis included 12 genes in the cubilin-megalin multiligand endocytic receptor complex. Altered expression of these genes was validated by quantitative RT-PCR, and immunohistochemical analysis demonstrated that megalin protein expression disappeared from the visceral yolk sac of RFC1-/- embryos, while cubilin protein was widely misexpressed

  6. Petrous Apex Cephalocele: Report of Two Cases and Review of the Literature

    Energy Technology Data Exchange (ETDEWEB)

    Jeong, Bo Seong; Lee, Ghi Jai; Shim, Jae Chan; Leee, Jae Myeong; Nam, Mee Young; Kim, Ho Kyun [Dept. of Diagnostic Radiology, Seoul Paik Hospital, Injei University College of Medicine, Seoul (Korea, Republic of)

    2011-05-15

    A petrous apex cephalocele is a rare lesion of the petrous apex. It can be discovered incidentally or can cause a suite of clinical problems, such as trigeminal neuralgia or cerebrospinal fluid leakage. Although this lesion can be misinterpreted as a pathologic lesion, the characteristic radiologic features can provide the diagnostic clue for distinguishing these two lesions and avoid unnecessary treatment. Here we present CT and MRI finding of petrous apex cephalocele in two patients with review of the literature.

  7. Hair-Cell Mechanotransduction Persists in TRP Channel Knockout Mice.

    Science.gov (United States)

    Wu, Xudong; Indzhykulian, Artur A; Niksch, Paul D; Webber, Roxanna M; Garcia-Gonzalez, Miguel; Watnick, Terry; Zhou, Jing; Vollrath, Melissa A; Corey, David P

    2016-01-01

    Members of the TRP superfamily of ion channels mediate mechanosensation in some organisms, and have been suggested as candidates for the mechanotransduction channel in vertebrate hair cells. Some TRP channels can be ruled out based on lack of an inner ear phenotype in knockout animals or pore properties not similar to the hair-cell channel. Such studies have excluded Trpv4, Trpa1, Trpml3, Trpm1, Trpm3, Trpc1, Trpc3, Trpc5, and Trpc6. However, others remain reasonable candidates. We used data from an RNA-seq analysis of gene expression in hair cells as well as data on TRP channel conductance to narrow the candidate group. We then characterized mice lacking functional Trpm2, Pkd2, Pkd2l1, Pkd2l2 and Pkd1l3, using scanning electron microscopy, auditory brainstem response, permeant dye accumulation, and single-cell electrophysiology. In all of these TRP-deficient mice, and in double and triple knockouts, mechanotransduction persisted. Together with published studies, these results argue against the participation of any of the 33 mouse TRP channels in hair cell transduction. PMID:27196058

  8. Hair-Cell Mechanotransduction Persists in TRP Channel Knockout Mice.

    Directory of Open Access Journals (Sweden)

    Xudong Wu

    Full Text Available Members of the TRP superfamily of ion channels mediate mechanosensation in some organisms, and have been suggested as candidates for the mechanotransduction channel in vertebrate hair cells. Some TRP channels can be ruled out based on lack of an inner ear phenotype in knockout animals or pore properties not similar to the hair-cell channel. Such studies have excluded Trpv4, Trpa1, Trpml3, Trpm1, Trpm3, Trpc1, Trpc3, Trpc5, and Trpc6. However, others remain reasonable candidates. We used data from an RNA-seq analysis of gene expression in hair cells as well as data on TRP channel conductance to narrow the candidate group. We then characterized mice lacking functional Trpm2, Pkd2, Pkd2l1, Pkd2l2 and Pkd1l3, using scanning electron microscopy, auditory brainstem response, permeant dye accumulation, and single-cell electrophysiology. In all of these TRP-deficient mice, and in double and triple knockouts, mechanotransduction persisted. Together with published studies, these results argue against the participation of any of the 33 mouse TRP channels in hair cell transduction.

  9. Galaxy cluster scaling relations measured with APEX-SZ

    CERN Document Server

    Bender, A N; Ade, P A R; Basu, K; Bertoldi, F; Burkutean, S; Clarke, J; Dahlin, D; Dobbs, M; Ferrusca, D; Flanigan, D; Halverson, N W; Holzapfel, W L; Horellou, C; Johnson, B R; Kermish, Z D; Klein, M; Kneissl, R; Lanting, T; Lee, A T; Mehl, J; Menten, K M; Muders, D; Nagarajan, A; Pacaud, F; Reichardt, C L; Richards, P L; Schaaf, R; Schwan, D; Sommer, M W; Spieler, H; Tucker, C; Westbrook, B

    2014-01-01

    We present thermal Sunyaev-Zel'dovich effect (SZE) measurements for 42 galaxy clusters observed at 150 GHz with the APEX-SZ experiment. For each cluster, we model the pressure profile and calculate the integrated Comptonization $Y$ to estimate the total thermal energy of the intracluster medium (ICM). We compare the measured $Y$ values to X-ray observables of the ICM from the literature (cluster gas mass $M_{gas}$, temperature $T_X$, and $Y_X =M_{gas}T_X$) that relate to total cluster mass. We measure power law scaling relations, including an intrinsic scatter, between the SZE and X-ray observables for both the X-ray selected and uniform REFLEX-DXL cluster sample and the full ad hoc APEX-SZ sample. We observe that the lack of uniform X-ray analysis for the full cluster sample introduces significant variability into the measured scaling relations and dominates the level of intrinsic scatter. For the REFLEX-DXL sample, we find results consistent with a self-similar model of cluster evolution dominated by gravit...

  10. Hearing results in surgery for primary petrous apex lesions.

    Science.gov (United States)

    Gianoli, G J; Amedee, R G

    1994-09-01

    Hearing preservation is a frequently mentioned phrase in the growing field of skull base surgery. Many authors have attempted to identify prognostic factors for successful hearing preservation, and many have suggested alternative procedures for preserving serviceable hearing. Few have mentioned hearing improvement with skull base surgical procedures. In this article we present the hearing results of 25 surgical procedures for primary petrous apex lesions. These include 13 cholesterol granulomas, 5 cholesteatomas, 4 mucoceles, and 3 eosinophilic granulomas. Surgical approaches included 14 transmastoid/infralabyrinthine, 6 transphenoid, 3 suboccipital, and 2 transmastoid/translabyrinthine. Hearing was maintained in 14 patients (56%), improved in 9 patients (36%), and worse in 1 patient with nonserviceable hearing before surgery (4%); 1 patient had profound hearing loss before surgery (4%). Results of this review should have significant implications on the choice of surgical approach for petrous apex lesions. Additionally, the standard method of determining salvageable hearing for most skull base procedures may not apply for this specific group of lesions. Implications for future treatment plans will be discussed in detail. PMID:8084633

  11. Metabolomic Characterization of Knockout Mutants in Arabidopsis: Development of a Metabolite Profiling Database for Knockout Mutants in Arabidopsis.

    Science.gov (United States)

    Fukushima, Atsushi; Kusano, Miyako; Mejia, Ramon Francisco; Iwasa, Mami; Kobayashi, Makoto; Hayashi, Naomi; Watanabe-Takahashi, Akiko; Narisawa, Tomoko; Tohge, Takayuki; Hur, Manhoi; Wurtele, Eve Syrkin; Nikolau, Basil J; Saito, Kazuki

    2014-05-14

    Despite recent intensive research efforts in functional genomics, the functions of only a limited number of Arabidopsis (Arabidopsis thaliana) genes have been determined experimentally, and improving gene annotation remains a major challenge in plant science. As metabolite profiling can characterize the metabolomic phenotype of a genetic perturbation in the plant metabolism, it provides clues to the function(s) of genes of interest. We chose 50 Arabidopsis mutants, including a set of characterized and uncharacterized mutants, that resemble wild-type plants. We performed metabolite profiling of the plants using gas chromatography-mass spectrometry. To make the data set available as an efficient public functional genomics tool for hypothesis generation, we developed the Metabolite Profiling Database for Knock-Out Mutants in Arabidopsis (MeKO). It allows the evaluation of whether a mutation affects metabolism during normal plant growth and contains images of mutants, data on differences in metabolite accumulation, and interactive analysis tools. Nonprocessed data, including chromatograms, mass spectra, and experimental metadata, follow the guidelines set by the Metabolomics Standards Initiative and are freely downloadable. Proof-of-concept analysis suggests that MeKO is highly useful for the generation of hypotheses for genes of interest and for improving gene annotation. MeKO is publicly available at http://prime.psc.riken.jp/meko/.

  12. Shoot Apex Demand Determines Assimilate and Nutrients Partitioning and Nutrient-uptake Rate in Tobacco Plants

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Our previous experiment revealed that apex-removed plants have larger root systems but a lower K+-uptake rates than intact tobacco plants.Since the apex is not only e center of growth and metabolism,but also an important place of auxin synthesis and export,the aims of this study were to distinguish whether the apex demand or auxin synthesized in the apex regulates assimilate and nutrients partitioning within plant,and to explain the reason for the lower K+-uptake rate of the apex-ramoved plant.In comparison with the control plant,covering the shoot apex with a black transparent plastic bag reduced net increases In dry matter and nutrients;however,the distribution of the dry matter and nutrients between shoot and roots and nutrient-uptake rates were not changed.Removal of the shoot apex shifted the dry mass and nutrients distributions to roots,and reduced the rate of nutrient uptake.Application of 1-naphthylacetic acid(NAA) could partly replace the role of the removed apex,stimulated assimilate and nutrient deposition into the treated tissue,and enhanced the reduced plasma membrane ATPase activity of roots to the control level.However,treatment of the apex-removed plants with NAA could not rescue the reduced nutrient uptake rate and the shifted assimilates and nutrients partitioning caused by excision of the apex.Higher nutrient uptake rate of the intact plants could not be explained by root growth parameters,such as total root surface area and number of root tips.The results from the present study indicate that strong apex demand determined assimilatas and nutrients partitioning and nutrient-uptake rate in tobacco(Nicotiana tabacum)plants.

  13. Modeling Halophytic Plants in APEX for Sustainable Water and Agriculture

    Science.gov (United States)

    DeRuyter, T.; Saito, L.; Nowak, B.; Rossi, C.; Toderich, K.

    2013-12-01

    A major problem for irrigated agricultural production is soil salinization, which can occur naturally or can be human-induced. Human-induced, or secondary salinization, is particularly a problem in arid and semi-arid regions, especially in irrigated areas. Irrigated land has more than twice the production of rainfed land, and accounts for about one third of the world's food, but nearly 20% of irrigated lands are salt-affected. Many farmers worldwide currently seasonally leach their land to reduce the soil salt content. These practices, however, create further problems such as a raised groundwater table, and salt, fertilizer, and pesticide pollution of nearby lakes and groundwater. In Uzbekistan, a combination of these management practices and a propensity to cultivate 'thirsty' crops such as cotton has also contributed to the Aral Sea shrinking nearly 90% by volume since the 1950s. Most common agricultural crops are glycophytes that have reduced yields when subjected to salt-stress. Some plants, however, are known as halophytic or 'salt-loving' plants and are capable of completing their life-cycle in higher saline soil or water environments. Halophytes may be useful for human consumption, livestock fodder, or biofuel, and may also be able to reduce or maintain salt levels in soil and water. To assess the potential for these halophytes to assist with salinity management, we are developing a model that is capable of tracking salinity under different management practices in agricultural environments. This model is interdisciplinary as it combines fields such as plant ecology, hydrology, and soil science. The US Department of Agriculture (USDA) model, Agricultural Policy/Environmental Extender (APEX), is being augmented with a salinity module that tracks salinity as separate ions across the soil-plant-water interface. The halophytes Atriplex nitens, Climacoptera lanata, and Salicornia europaea are being parameterized and added into the APEX model database. Field sites

  14. Establishment, identification and preliminary phenotype of mouse model of specific Smad4 conditional gene knockout in lens ectoderm%条件性 Sm ad4基因敲除鼠模型的建立和鉴定及初步表型

    Institute of Scientific and Technical Information of China (English)

    刘瑛; 林丁

    2014-01-01

    analysis by establishing the mouse model of specific Smad4 conditional gene knockout in ocular tissue by Cre/LoxP system of this kind of mouse model. METHODS: Mouse of specific Smad4 conditional knockout in lens ectoderm ( Le-Cre; Samd4fl/fl or also called mutant mouse) was obtained by mating the Pax6 promoter-driven Cre transgenic mouse ( Le-Cre ) with Smad4 wildtype mouse ( Smad4 fl/fl).To confirm that Smad4 has been conditionally inactivated only in the specific tissue of ectoderm such as lens, cornea and ectoderm of the eyelids so on.A series of assays were carried out to reveal the validity and specificity of Smad4 gene knockout at molecular and cellular levels, including genotyping by PCR, detection of green fluorescence protein ( GFP) in specific tissue and Smad4 protein.The expression of Le-Cre from Lacz staining using ROSA26 reporter genes in specific ocular tissue of mice can be visualized.Preliminary phenotype of mutant mouse was also observed. RESULTS: As early as around E10.0, strong GFP expression was observed in the embryonic lens and periorbital ectoderm of the mice, which showed Le-Cre was expressed in specific target tissue. Through genotyping for Smad4, Cre and Rosa genes, the mice were determined if they have carried Cre, Smad4 allele or Rosa reporter gene.It was further confirmed by lens-sampled genotyping that Smad4 gene was removed from some specific tissue such as lens.The spatial-temporal expression and tissue specificity of Le-Cre recombinase was also revealed by LacZ staining of Rosa; Le-Cre double transgenic mouse. According to Immunohistochemical staining, Smad4 was widely expressed in normal embryonic eyes, mainly appearing in the cytoplasm at the early embryonic stage and were transferred to nucleus with gestation developing, while in mutant embryonic eyes, Smad4 was void of expression in Cre-expressed tissues. It was observed that Smad4 mutant mouse could survive the conditional gene knockout.But those mice showed abnormal appearance such

  15. The Expression of TALEN before Fertilization Provides a Rapid Knock-Out Phenotype in Xenopus laevis Founder Embryos.

    Directory of Open Access Journals (Sweden)

    Kei Miyamoto

    Full Text Available Recent advances in genome editing using programmable nucleases have revolutionized gene targeting in various organisms. Successful gene knock-out has been shown in Xenopus, a widely used model organism, although a system enabling less mosaic knock-out in founder embryos (F0 needs to be explored in order to judge phenotypes in the F0 generation. Here, we injected modified highly active transcription activator-like effector nuclease (TALEN mRNA to oocytes at the germinal vesicle (GV stage, followed by in vitro maturation and intracytoplasmic sperm injection, to achieve a full knock-out in F0 embryos. Unlike conventional injection methods to fertilized embryos, the injection of TALEN mRNA into GV oocytes allows expression of nucleases before fertilization, enabling them to work from an earlier stage. Using this procedure, most of developed embryos showed full knock-out phenotypes of the pigmentation gene tyrosinase and/or embryonic lethal gene pax6 in the founder generation. In addition, our method permitted a large 1 kb deletion. Thus, we describe nearly complete gene knock-out phenotypes in Xenopus laevis F0 embryos. The presented method will help to accelerate the production of knock-out frogs since we can bypass an extra generation of about 1 year in Xenopus laevis. Meantime, our method provides a unique opportunity to rapidly test the developmental effects of disrupting those genes that do not permit growth to an adult able to reproduce. In addition, the protocol shown here is considerably less invasive than the previously used host transfer since our protocol does not require surgery. The experimental scheme presented is potentially applicable to other organisms such as mammals and fish to resolve common issues of mosaicism in founders.

  16. Effect of knockout of α2δ-1 on action potentials in mouse sensory neurons.

    Science.gov (United States)

    Margas, Wojciech; Ferron, Laurent; Nieto-Rostro, Manuela; Schwartz, Arnold; Dolphin, Annette C

    2016-08-01

    Gene deletion of the voltage-gated calcium channel auxiliary subunit α2δ-1 has been shown previously to have a cardiovascular phenotype, and a reduction in mechano- and cold sensitivity, coupled with delayed development of neuropathic allodynia. We have also previously shown that dorsal root ganglion (DRG) neuron calcium channel currents were significantly reduced in α2δ-1 knockout mice. To extend our findings in these sensory neurons, we have examined here the properties of action potentials (APs) in DRG neurons from α2δ-1 knockout mice in comparison to their wild-type (WT) littermates, in order to dissect how the calcium channels that are affected by α2δ-1 knockout are involved in setting the duration of individual APs and their firing frequency. Our main findings are that there is reduced Ca(2+) entry on single AP stimulation, particularly in the axon proximal segment, reduced AP duration and reduced firing frequency to a 400 ms stimulation in α2δ-1 knockout neurons, consistent with the expected role of voltage-gated calcium channels in these events. Furthermore, lower intracellular Ca(2+) buffering also resulted in reduced AP duration, and a lower frequency of AP firing in WT neurons, mimicking the effect of α2δ-1 knockout. By contrast, we did not obtain any consistent evidence for the involvement of Ca(2+)-activation of large conductance calcium-activated potassium (BK) and small conductance calcium-activated potassium (SK) channels in these events. In conclusion, the reduced Ca(2+) elevation as a result of single AP stimulation is likely to result from the reduced duration of the AP in α2δ-1 knockout sensory neurons.This article is part of the themed issue 'Evolution brings Ca(2+) and ATP together to control life and death'. PMID:27377724

  17. Gain and frequency tuning within the mouse cochlear apex

    International Nuclear Information System (INIS)

    Normal mammalian hearing requires cochlear outer hair cell active processes that amplify the traveling wave with high gain and sharp tuning, termed cochlear amplification. We have used optical coherence tomography to study cochlear amplification within the apical turn of the mouse cochlea. We measured not only classical basilar membrane vibratory tuning curves but also vibratory responses from the rest of the tissues that compose the organ of Corti. Basilar membrane tuning was sharp in live mice and broad in dead mice, whereas other regions of the organ of Corti demonstrated phase shifts consistent with additional filtering beyond that provided by basilar membrane mechanics. We use these experimental data to support a conceptual framework of how cochlear amplification is tuned within the mouse cochlear apex. We will also study transgenic mice with targeted mutations that affect different biomechanical aspects of the organ of Corti in an effort to localize the underlying processes that produce this additional filtering

  18. Do neonatal mouse hearts regenerate following heart apex resection?

    DEFF Research Database (Denmark)

    Andersen, Ditte Caroline; Ganesalingam, Suganya; Jensen, Charlotte Harken;

    2014-01-01

    strains and found no evidence of complete regeneration. Ideally, new functional cardiomyocytes, endothelial cells, and vascular smooth muscle cells should be formed in the necrotic area of the damaged heart. Here, damaged hearts were 9.8% shorter and weighed 14% less than sham controls. In addition......The mammalian heart has generally been considered nonregenerative, but recent progress suggests that neonatal mouse hearts have a genuine capacity to regenerate following apex resection (AR). However, in this study, we performed AR or sham surgery on 400 neonatal mice from inbred and outbred......, the resection border contained a massive fibrotic scar mainly composed of nonmyocytes and collagen disposition. Furthermore, there was a substantial reduction in the number of proliferating cardiomyocytes in AR hearts. Our results thus question the usefulness of the AR model for identifying molecular mechanisms...

  19. Low altitude dose measurements from APEX, CRRES and DMSP.

    Science.gov (United States)

    Mullen, E G; Gussenhoven, M S; Bell, J T; Madden, D; Holeman, E; Delorey, D

    1998-01-01

    Dosimeter data taken on the APEX (1994-1996), CRRES (1990-1991) and DMSP (1984-1987) satellites have been used to study the low altitude (down to 350 km) radiation environment. Of special concern has been the inner edge of the inner radiation belt due to its steep gradient. We have constructed dose models of the inner edge of the belt from all three spacecraft and put them into a personal computer utility, called APEXRAD, that calculates dose for user-selected orbits. The variation of dose for low altitude, circular orbits is given as a function of altitude, inclination and particle type. Dose-depth curves show that shielding greater than approximately 1/4 in Al is largely ineffectual for low altitude orbits. The contribution of outer zone electrons to low altitude dose is shown to be important only for thin shields and to have significant variation with magnetic activity and solar cycle.

  20. Gain and frequency tuning within the mouse cochlear apex

    Energy Technology Data Exchange (ETDEWEB)

    Oghalai, John S.; Raphael, Patrick D. [Department of Otolaryngology, Stanford University School of Medicine, Stanford, California (United States); Gao, Simon [Department of Otolaryngology, Stanford University School of Medicine, Stanford, California (United States); Department of Bioengineering, Rice University, Houston, Texas (United States); Lee, Hee Yoon [Department of Otolaryngology, Stanford University School of Medicine, Stanford, California (United States); Department of Electrical Engineering, Stanford University, Stanford, California (United States); Groves, Andrew K. [Department of Neuroscience, Department of Molecular and Human Genetics, and Program in Developmental Biology, Baylor College of Medicine, Houston, Texas (United States); Zuo, Jian [Department of Developmental Neurobiology, St. Jude Children’s Research Hospital, Memphis, Tennessee (United States); Applegate, Brian E. [Department of Biomedical Engineering, Texas A& M University, College Station, Texas (United States)

    2015-12-31

    Normal mammalian hearing requires cochlear outer hair cell active processes that amplify the traveling wave with high gain and sharp tuning, termed cochlear amplification. We have used optical coherence tomography to study cochlear amplification within the apical turn of the mouse cochlea. We measured not only classical basilar membrane vibratory tuning curves but also vibratory responses from the rest of the tissues that compose the organ of Corti. Basilar membrane tuning was sharp in live mice and broad in dead mice, whereas other regions of the organ of Corti demonstrated phase shifts consistent with additional filtering beyond that provided by basilar membrane mechanics. We use these experimental data to support a conceptual framework of how cochlear amplification is tuned within the mouse cochlear apex. We will also study transgenic mice with targeted mutations that affect different biomechanical aspects of the organ of Corti in an effort to localize the underlying processes that produce this additional filtering.

  1. Generation of transient receptor potential vanilloid 6 gene knockout mouse model%瞬时性受体电位通道香草酸受体亚型6基因敲除小鼠模型的建立

    Institute of Scientific and Technical Information of China (English)

    叶添文; 郑晋华; 安静; 郭清河; 陈方经; 陈爱民

    2012-01-01

    目的 建立瞬时性受体电位通道香草酸受体亚型6(Trpv6)基因敲除小鼠模型,为在体研究Trpv6的生物学功能及其与骨代谢关系奠定基础.方法 从Ensembl数据库中获得小鼠Trpv6基因组序列.设计基因敲除策略,构建基因敲除载体pBR322-MK-Trpv6.以电穿孔方法将基因敲除载体导入胚胎多能干细胞(ES细胞),用G418和Ganciclovoir进行正负筛选,获得双抗性克隆.PCR鉴定出正确同源重组的ES细胞克隆.将正确同源重组的ES细胞注射到C57BL/6J小鼠的囊胚中,获得嵌合体小鼠.挑选嵌合率在50%的雄鼠与C57BL/6J小鼠交配,获得的灰鼠经PCR鉴定为杂合子小鼠.杂合子小鼠交配后获得纯合子小鼠.结果 成功构建了打靶载体pBR322-MK-Trpv6.电穿孔后,共获得24个正确同源重组的克隆,同源重组效率为25%.同源重组的克隆经显微注射后,共获得4只嵌合率大于50%的雄鼠.嵌合鼠与C57BL/6J小鼠交配,获得57只来源于ES细胞的灰鼠,PCR鉴定证实其中17只为杂合子小鼠,阳性率为29.8%.杂合子小鼠交配获得纯合子小鼠.经蛋白质印迹分析证实纯合子小鼠无Trpv6蛋白的表达.结论 成功建立了Trpv6基因敲除小鼠模型,其中纯合子小鼠未出现胚胎致死现象.%Objective To create transient receptor potential vanilloid 6 (Trpv6) gene knockout mouse model, so as to pave a way for further research of its biological function and its role in bone metabolism in vivo. Methods Mouse genomic DNA sequence of Trpv6 gene was obtained from Ensembl database. Trpv6 gene knockout vector (pBR322-MK-Trpv6) was constructed. Trpv6 knockout vector was transferred into the embryonic stem (ES) cells by electroporation and screening of both G418 and Ganciclovoir resistant clones were performed routinely. The homologous recombined ES cell clones were identified by PCR. The correct homologously recombined ES cells were microinjected into C57BL/6J mouse blastocysts to obtain chimera

  2. L1CAM基因敲除小鼠的脑局部血流量、ATP含量和蛋白合成率分析%Analysis of focal cerebral blood flow, ATP content and protein sythesis of L1 gene knockout mice

    Institute of Scientific and Technical Information of China (English)

    张玲; Hossmann Alex

    2007-01-01

    目的 对L1细胞黏附分子(L1 cell adhesion molecule,L1CAM)基因敲除(gene knockout,KO)小鼠的各脑区脑局部血流量、ATP含量和蛋白合成率进行分析,探讨L1CAM基因缺失对小鼠脑局部代谢功能有无影响.方法 用放射性自显影方法测定L1 KO小鼠和野生型对照组小鼠大脑局部血流量和蛋白质合成率,用生物荧光影像技术测定两组ATP含量,并将两组数据进行比较.结果 与野生型小鼠相比,L1 KO小鼠局部脑血流量、ATP含量和蛋白质合成率差异均无统计学意义(均P>0.05).结论 L1CAM基因缺失对小鼠脑局部代谢功能无明显影响,提示L1 KO小鼠可作为研究L1CAM在脑缺血中作用的动物模型.

  3. Growth plate closure: Apex view on bone scan

    International Nuclear Information System (INIS)

    Angular deformities of the extremities in children following premature closure of the growth plate are well known. The deformities depend on the position of an osseus bridge which forms between the epiphysis and metaphysis. Several surgical procedures including resection of the osseus bridge have been described, however, delineation of the site of fusion is difficult to define. The commonest site of growth plate arrest is the distal femoral or proximal tibial growth plate. A new technique using the bone scan has been developed which accurately defines the area and position of these osseus bridges. Two hours after injection of technetium 99m methylene diphosphonate apex views of the affected distal femoral growth plate were performed. The knee was flexed into its smallest angle. Using a pinhole collimator the gamma camera was angled to face the affected growth plate end on. The image was collected onto computer and analysed by: (I) regions of interest over segments of the growth plate to calculate the relative area of total growth plate affected: (II) generating histograms: (III) thresholding or performing isocontours to accentuate abnormal areas. The growth plate is normally uniformly increased when compared to the normal shaft of the bone. Fusion across the plate appears as an area of diminished uptake. The apex view gives a unique functional map of the growth plate such that abnormal areas are displayed, and the site, size and position of osseus fusion obtained. The technique has the potential for determining the metabolic activity of the growth plate before and after surgery. Serial studies will allow assessment of regneration of the plate and reformation of new osseus bridges

  4. METRO-APEX Volume 13.1: Industrialist's Manual No. 3, Rusty's Iron Foundry. Revised.

    Science.gov (United States)

    University of Southern California, Los Angeles. COMEX Research Project.

    The Industrialist's Manual No. 3 (Rusty's Iron Foundry) is one of a set of twenty-one manuals used in METRO-APEX 1974, a computerized college and professional level, computer-supported, role-play, simulation exercise of a community with "normal" problems. Stress is placed on environmental quality considerations. APEX 1974 is an expansion of…

  5. APEX simulation: environmental benefits of agroforestry and grass buffers on corn-soybean watersheds

    Science.gov (United States)

    The Agricultural Policy Environmental Extender (APEX) model has the ability to simulate the effects of vegetative filter strips on runoff and pollutant loadings from agricultural watersheds. The objectives of this study were to calibrate and validate the APEX model for three adjacent watersheds and...

  6. 75 FR 82335 - Airworthiness Directives; APEX Aircraft Model CAP 10 Airplanes

    Science.gov (United States)

    2010-12-30

    ... identified in this proposed AD, contact Apex Aircraft, Bureau de Navigabilit , 1 route de Troyes, 21121... route de Troyes, 21121 DAROIS- France, telephone: (33) 380 35 65 10; fax: (33) 380 35 65 15; email: apex... rule'' under the DOT Regulatory Policies and Procedures (44 FR 11034, February 26, 1979); and 3....

  7. Root cap-dependent gravitropic U-turn of maize root requires light-induced auxin biosynthesis via the YUC pathway in the root apex.

    Science.gov (United States)

    Suzuki, Hiromi; Yokawa, Ken; Nakano, Sayuri; Yoshida, Yuriko; Fabrissin, Isabelle; Okamoto, Takashi; Baluška, František; Koshiba, Tomokazu

    2016-08-01

    Gravitropism refers to the growth or movement of plants that is influenced by gravity. Roots exhibit positive gravitropism, and the root cap is thought to be the gravity-sensing site. In some plants, the root cap requires light irradiation for positive gravitropic responses. However, the mechanisms regulating this phenomenon are unknown. We herein report that maize roots exposed to white light continuously for ≥1-2h show increased indole-3-acetic acid (IAA) levels in the root tips, especially in the transition zone (1-3mm from the tip). Treatment with IAA biosynthesis inhibitors yucasin and l-kynurenine prevented any increases in IAA content and root curvature under light conditions. Analyses of the incorporation of a stable isotope label from tryptophan into IAA revealed that some of the IAA in roots was synthesized in the root apex. Furthermore, Zmvt2 and Zmyuc gene transcripts were detected in the root apex. One of the Zmyuc genes (ZM2G141383) was up-regulated by light irradiation in the 0-1mm tip region. Our findings suggest that IAA accumulation in the transition zone is due to light-induced activation of Zmyuc gene expression in the 0-1mm root apex region. Light-induced changes in IAA levels and distributions mediate the maize root gravitropic U-turn. PMID:27307546

  8. The Influence of Knockout ofmenA Gene inEscherichia coli onthe Accumulation of CoQ%大肠杆菌menA敲除对CoQ积累的影响研究

    Institute of Scientific and Technical Information of China (English)

    刘永青; 任琳; 张子锋

    2015-01-01

    通过同源重组敲除大肠杆菌的menA基因,增加菌体CoQ合成量,用于构建CoQ高产菌株。以pKD4质粒为模板, PCR扩增kanr片段;在pKD46的辅助下,kanr片段转化大肠杆菌,利用抗生素筛选和PCR验证重组子;以紫外诱变菌为对照,发酵menA基因敲除菌株,分析CoQ种类和产量变化。成功获得menA基因敲除菌株,CoQ种类不变,产量增加约为38%。首次敲除menA基因,改良株CoQ产量得到提高,达到预期目标。%Knocking outmenA gene of Escherichia coli by homologous recombination increases the synthetized amount of CoQ, which is used for constructing the strains of high-yield CoQ. Using pKD4 plasmid as template, thekanr fragments were amplified by PCR. In the presence of auxiliary plasmid pKD46, thekanr fragments were transformed intoEscherichia coli, and the recombinant one was confirmed by antibiotic screening and verification of PCR;By contrast with uv-induced mutation, the strains withmenA gene knocked out were fermented, and the types of CoQ and the changes of CoQ yield were analyzed. The results showed that thestrains withmenAgeneknockout were successfully obtained, while the types of CoQ unchanged and the yield increased about 38%. In conclusion, this is the first time of knocking outmenA gene, CoQ production of mutant strains is improved, and the desired goal is achieved, which lays a foundation for the further construction of high-yield CoQ strains.

  9. CRED APEX Drifting Buoy Argos_ID 25321 Data in the Northwestern Hawaiian Islands, 200110-200404 (NODC Accession 0049436)

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — CRED APEX drifter Argos_ID 25321 was deployed in the region of NW Hawaiian Islands to assess ocean currents and sea surface temperature. APEX drifter data files...

  10. CRED APEX Drifting Buoy Argos_ID 25330 Data Maro Reef, Northwestern Hawaiian Islands, 200109-200201 (NODC Accession 0049436)

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — CRED APEX drifter Argos_ID 25330 was deployed in the region of NW Hawaiian Islands to assess ocean currents and sea surface temperature. APEX drifter data files...

  11. CRED APEX Drifting Buoy Argos_ID 25333 Data in the NW Hawaiian Islands, 200110-200204 (NODC Accession 0049436)

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — CRED APEX drifter Argos_ID 25333 was deployed in the region of NW Hawaiian Islands to assess ocean currents and sea surface temperature. APEX drifter data files...

  12. Sdhd and SDHD/H19 knockout mice do not develop paraganglioma or pheochromocytoma.

    Directory of Open Access Journals (Sweden)

    Jean-Pierre Bayley

    Full Text Available BACKGROUND: Mitochondrial succinate dehydrogenase (SDH is a component of both the tricarboxylic acid cycle and the electron transport chain. Mutations of SDHD, the first protein of intermediary metabolism shown to be involved in tumorigenesis, lead to the human tumors paraganglioma (PGL and pheochromocytoma (PC. SDHD is remarkable in showing an 'imprinted' tumor suppressor phenotype. Mutations of SDHD show a very high penetrance in man and we postulated that knockout of Sdhd would lead to the development of PGL/PC, probably in aged mice. METHODOLOGY/PRINCIPAL FINDINGS: We generated a conventional knockout of Sdhd in the mouse, removing the entire third exon. We also crossed this mouse with a knockout of H19, a postulated imprinted modifier gene of Sdhd tumorigenesis, to evaluate if loss of these genes together would lead to the initiation or enhancement of tumor development. Homozygous knockout of Sdhd results in embryonic lethality. No paraganglioma or other tumor development was seen in Sdhd KO mice followed for their entire lifespan, in sharp contrast to the highly penetrant phenotype in humans. Heterozygous Sdhd KO mice did not show hyperplasia of paraganglioma-related tissues such as the carotid body or of the adrenal medulla, or any genotype-related pathology, with similar body and organ weights to wildtype mice. A cohort of Sdhd/H19 KO mice developed several cases of profound cardiac hypertrophy, but showed no evidence of PGL/PC. CONCLUSIONS: Knockout of Sdhd in the mouse does not result in a disease phenotype. H19 may not be an initiator of PGL/PC tumorigenesis.

  13. Single-Step Generation of Conditional Knockout Mouse Embryonic Stem Cells

    Directory of Open Access Journals (Sweden)

    Matyas Flemr

    2015-07-01

    Full Text Available Induction of double-strand DNA breaks (DSBs by engineered nucleases, such as CRISPR/Cas9 or transcription activator-like effector nucleases (TALENs, stimulates knockin of exogenous DNA fragments via homologous recombination (HR. However, the knockin efficiencies reported so far have not allowed more complex in vitro genome modifications such as, for instance, simultaneous integration of a DNA fragment at two distinct genomic sites. We developed a reporter system to enrich for cells with engineered nuclease-assisted HR events. Using this system in mouse embryonic stem cells (mESCs, we achieve single-step biallelic and seamless integration of two loxP sites for Cre recombinase-mediated inducible gene knockout, as well as biallelic endogenous gene tagging with high efficiency. Our approach reduces the time and resources required for conditional knockout mESC generation dramatically.

  14. Universal statistics of the knockout tournament

    Science.gov (United States)

    Baek, Seung Ki; Yi, Il Gu; Park, Hye Jin; Kim, Beom Jun

    2013-11-01

    We study statistics of the knockout tournament, where only the winner of a fixture progresses to the next. We assign a real number called competitiveness to each contestant and find that the resulting distribution of prize money follows a power law with an exponent close to unity if the competitiveness is a stable quantity and a decisive factor to win a match. Otherwise, the distribution is found narrow. The existing observation of power law distributions in various kinds of real sports tournaments therefore suggests that the rules of those games are constructed in such a way that it is possible to understand the games in terms of the contestants' inherent characteristics of competitiveness.

  15. Universal statistics of the knockout tournament

    CERN Document Server

    Baek, Seung Ki; Park, Hye Jin; Kim, Beom Jun

    2014-01-01

    We study statistics of the knockout tournament, where only the winner of a fixture progresses to the next. We assign a real number called competitiveness to each contestant and find that the resulting distribution of prize money follows a power law with an exponent close to unity if the competitiveness is a stable quantity and a decisive factor to win a match. Otherwise, the distribution is found narrow. The existing observation of power law distributions in various kinds of real sports tournaments therefore suggests that the rules of those games are constructed in such a way that it is possible to understand the games in terms of the contestants' inherent characteristics of competitiveness.

  16. Histidine decarboxylase knockout mice, a genetic model of Tourette syndrome, show repetitive grooming after induced fear.

    Science.gov (United States)

    Xu, Meiyu; Li, Lina; Ohtsu, Hiroshi; Pittenger, Christopher

    2015-05-19

    Tics, such as are seen in Tourette syndrome (TS), are common and can cause profound morbidity, but they are poorly understood. Tics are potentiated by psychostimulants, stress, and sleep deprivation. Mutations in the gene histidine decarboxylase (Hdc) have been implicated as a rare genetic cause of TS, and Hdc knockout mice have been validated as a genetic model that recapitulates phenomenological and pathophysiological aspects of the disorder. Tic-like stereotypies in this model have not been observed at baseline but emerge after acute challenge with the psychostimulant d-amphetamine. We tested the ability of an acute stressor to stimulate stereotypies in this model, using tone fear conditioning. Hdc knockout mice acquired conditioned fear normally, as manifested by freezing during the presentation of a tone 48h after it had been paired with a shock. During the 30min following tone presentation, knockout mice showed increased grooming. Heterozygotes exhibited normal freezing and intermediate grooming. These data validate a new paradigm for the examination of tic-like stereotypies in animals without pharmacological challenge and enhance the face validity of the Hdc knockout mouse as a pathophysiologically grounded model of tic disorders.

  17. Transgenic knockout mice with exclusively human sickle hemoglobinand sickle cell disease

    Energy Technology Data Exchange (ETDEWEB)

    Paszty, C.; Brion, C.; Manci, E.; Witkowska, E.; Stevens, M.; Narla, M.; Rubin, E.

    1997-06-13

    To create mice expressing exclusively human sicklehemoglobin (HbS), transgenic mice expressing human alpha-, gamma-, andbeta[S]-globin were generated and bred with knockout mice that haddeletions of the murine alpha- and beta-globin genes. These sickle cellmice have the major features (irreversibly sickled red cells, anemia,multiorgan pathology) found in humans with sickle cell disease and, assuch, represent a useful in vivo system to accelerate the development ofimproved therapies for this common genetic disease.

  18. Key Regulators of Mitochondrial Biogenesis are Increased in Kidneys of Growth Hormone Receptor Knockout (GHRKO) Mice

    OpenAIRE

    Gesing, Adam; Bartke, Andrzej; Wang, Feiya; Karbownik-Lewinska, Malgorzata; Masternak, Michal M.

    2011-01-01

    The growth hormone (GH) receptor knockout mice (GHRKO) are remarkably long-lived and highly insulin sensitive. Alterations in mitochondrial biogenesis are associated with aging and various metabolic derangements. We have previously demonstrated increased gene expression of key regulators of mitochondriogenesis in kidneys, hearts and skeletal muscles of GHRKO mice. The aim of the present study was to quantify the protein levels of the following regulators of mitochondriogenesis: peroxisome pro...

  19. Sirh7/Ldoc1 knockout mice exhibit placental P4 overproduction and delayed parturition

    OpenAIRE

    Naruse, Mie; Ono, Ryuichi; Irie, Masahito; Nakamura, Kenji; Furuse, Tamio; Hino, Toshiaki; Oda, Kanako; Kashimura, Misho; Yamada, Ikuko; Wakana, Shigeharu; Yokoyama, Minesuke; Ishino, Fumitoshi; Kaneko-Ishino, Tomoko

    2014-01-01

    Sirh7/Ldoc1 [sushi-ichi retrotransposon homolog 7/leucine zipper, downregulated in cancer 1, also called mammalian retrotransposon-derived 7 (Mart7)] is one of the newly acquired genes from LTR retrotransposons in eutherian mammals. Interestingly, Sirh7/Ldoc1 knockout (KO) mice exhibited abnormal placental cell differentiation/maturation, leading to an overproduction of placental progesterone (P4) and placental lactogen 1 (PL1) from trophoblast giant cells (TGCs). The placenta is an organ tha...

  20. Comprehensive reanalysis of transcription factor knockout expression data in Saccharomyces cerevisiae reveals many new targets

    OpenAIRE

    Reimand, Jüri; Vaquerizas, Juan M.; Todd, Annabel E.; Vilo, Jaak; Luscombe, Nicholas M.

    2010-01-01

    Transcription factor (TF) perturbation experiments give valuable insights into gene regulation. Genome-scale evidence from microarray measurements may be used to identify regulatory interactions between TFs and targets. Recently, Hu and colleagues published a comprehensive study covering 269 TF knockout mutants for the yeast Saccharomyces cerevisiae. However, the information that can be extracted from this valuable dataset is limited by the method employed to process the microarray data. Here...

  1. Altered reward circuitry in the norepinephrine transporter knockout mouse.

    Directory of Open Access Journals (Sweden)

    Joseph J Gallagher

    Full Text Available Synaptic levels of the monoamine neurotransmitters dopamine, serotonin, and norepinephrine are modulated by their respective plasma membrane transporters, albeit with a few exceptions. Monoamine transporters remove monoamines from the synaptic cleft and thus influence the degree and duration of signaling. Abnormal concentrations of these neuronal transmitters are implicated in a number of neurological and psychiatric disorders, including addiction, depression, and attention deficit/hyperactivity disorder. This work concentrates on the norepinephrine transporter (NET, using a battery of in vivo magnetic resonance imaging techniques and histological correlates to probe the effects of genetic deletion of the norepinephrine transporter on brain metabolism, anatomy and functional connectivity. MRS recorded in the striatum of NET knockout mice indicated a lower concentration of NAA that correlates with histological observations of subtle dysmorphisms in the striatum and internal capsule. As with DAT and SERT knockout mice, we detected minimal structural alterations in NET knockout mice by tensor-based morphometric analysis. In contrast, longitudinal imaging after stereotaxic prefrontal cortical injection of manganese, an established neuronal circuitry tracer, revealed that the reward circuit in the NET knockout mouse is biased toward anterior portions of the brain. This is similar to previous results observed for the dopamine transporter (DAT knockout mouse, but dissimilar from work with serotonin transporter (SERT knockout mice where Mn(2+ tracings extended to more posterior structures than in wildtype animals. These observations correlate with behavioral studies indicating that SERT knockout mice display anxiety-like phenotypes, while NET knockouts and to a lesser extent DAT knockout mice display antidepressant-like phenotypic features. Thus, the mainly anterior activity detected with manganese-enhanced MRI in the DAT and NET knockout mice is likely

  2. Recoiled Proton Tagged Knockout Reaction for 8He

    Institute of Scientific and Technical Information of China (English)

    曹中鑫; 叶沿林; 江栋兴; 郑涛; 李智焕; 华辉; 葛榆成; 李湘庆; 楼建玲; 肖军; 李奇特; 吕林辉; 李阔昂; 王赫; 乔锐; 游海波; 陈瑞九

    2012-01-01

    An experiment for knockout reaction induced by SHe beam at 82.5 MeV/nucleon on CH2 and C targets was carried out. The 6He and 4He core fragments at forward angles and the recoiled protons at large angles were detected coincidently. From this exclusive measurement the valence nucleon knockout mechanism and the core knockout mechanism are separated, which can be applied to the exclusive spectroscopic study on the structure of exotic nuclei.

  3. Knockout of the iha gene in uropathogenic Escherichia coli and its influence on biofilm formation%尿路致病性大肠杆菌iha基因敲除及其对细菌生物膜形成的影响

    Institute of Scientific and Technical Information of China (English)

    张晓雷; 毛立群; 葛新; 董小青

    2013-01-01

    Objective To construct the adhesin gene iha mutant of uropathogenic Escherichia coli (UPEC) strain W140,and analyze the effects of iha deletion on UPEC biofilm formation.Methods Knockout of the iha gene was done with plasmid pKD46,pKD3,and pCP20 of Red recombinant system.pKD46 was transformed into W140 strain to express the three recombination proteins of λ phage.The chloramphenicol resistance gene flanked by flipase recognition target (FRT) of pKD3 was amplified by PCR and transformed into W140 strain to replace the iha gene.Finally pCP20 was transformed into W140 strain and expressed flipase recombinase,which deleted the chloramphenicol resistance gene between FRT sites.The differences of biofilm formation ability between the wild strain and the mutant were measured by crystal violet staining.Results PCR verification and DNA sequencing indicated that the iha gene was successfully knocked out and UPEC W140Δiha was constructed.The biofilm growth level of the mutant was significantly lower than that of wild strain (P <0.01).Conclusion The iha gene and its encoding product are involved in the process of UPEC biofilm formation.Suppressing iha gene may provide a new target for controlling urinary tract infection.%目的 构建尿路致病性大肠杆菌(UPEC) W140菌株的黏附素基因iha缺陷株,分析iha基因缺失对UPEC生物膜形成的影响.方法 采用Red重组系统的3种质粒(pKD46、pKD3、pCP20)敲除W140菌株的iha基因.pKD46表达λ噬菌体的3个重组蛋白,转入W140菌株使其具有同源重组能力.以pKD3携带的两侧带有翻转酶结合位点(FRT)的氯霉素抗性基因替换目的基因iha,再利用表达翻转酶重组酶的质粒pCP20将FRT之间的氯霉素抗性基因删除,从而获得iha基因敲除菌株.采用结晶紫染色法比较野生菌株与基因敲除菌株的细菌生物膜形成差异.结果 PCR验证和DNA测序表明,UPEC W140菌株染色体上的iha基因被成功敲除,得到iha基因缺陷株UPEC W140

  4. Proteomic Analysis of Loricrin Knockout Mouse Epidermis.

    Science.gov (United States)

    Rice, Robert H; Durbin-Johnson, Blythe P; Ishitsuka, Yosuke; Salemi, Michelle; Phinney, Brett S; Rocke, David M; Roop, Dennis R

    2016-08-01

    The crosslinked envelope of the mammalian epidermal corneocyte serves as a scaffold for assembly of the lipid barrier of the epidermis. Thus, deficient envelope crosslinking by keratinocyte transglutaminase (TGM1) is a major cause of the human autosomal recessive congenital ichthyoses characterized by barrier defects. Expectations that loss of some envelope protein components would also confer an ichthyosis phenotype have been difficult to demonstrate. To help rationalize this observation, the protein profile of epidermis from loricrin knockout mice has been compared to that of wild type. Despite the mild phenotype of the knockout, some 40 proteins were incorporated into envelope material to significantly different extents compared to those of wild type. Nearly half were also incorporated to similarly altered extents into the disulfide bonded keratin network of the corneocyte. The results suggest that loss of loricrin alters their incorporation into envelopes as a consequence of protein-protein interactions during cell maturation. Mass spectrometric protein profiling revealed that keratin 1, keratin 10, and loricrin are prominent envelope components and that dozens of other proteins are also components. This finding helps rationalize the potential formation of functional envelopes, despite loss of a single component, due to the availability of many alternative transglutaminase substrates. PMID:27418529

  5. Cytochrome P450 1b1 in polycyclic aromatic hydrocarbon (PAH)-induced skin carcinogenesis: Tumorigenicity of individual PAHs and coal-tar extract, DNA adduction and expression of select genes in the Cyp1b1 knockout mouse

    Energy Technology Data Exchange (ETDEWEB)

    Siddens, Lisbeth K. [Department of Environmental and Molecular Toxicology, Oregon State University, Corvallis, OR 97331 (United States); Superfund Research Center, Oregon State University, Corvallis, OR 97331 (United States); Bunde, Kristi L. [College of Veterinary Medicine, Oregon State University, Corvallis, OR 97331 (United States); Harper, Tod A. [Department of Environmental and Molecular Toxicology, Oregon State University, Corvallis, OR 97331 (United States); Linus Pauling Institute, Oregon State University, Corvallis, OR 97331 (United States); Environmental Health Sciences Center, Oregon State University, Corvallis, OR 97331 (United States); McQuistan, Tammie J. [Superfund Research Center, Oregon State University, Corvallis, OR 97331 (United States); Linus Pauling Institute, Oregon State University, Corvallis, OR 97331 (United States); Löhr, Christiane V. [Environmental Health Sciences Center, Oregon State University, Corvallis, OR 97331 (United States); College of Veterinary Medicine, Oregon State University, Corvallis, OR 97331 (United States); Bramer, Lisa M. [Applied Statistics and Computational Modeling, Pacific Northwest National Laboratory, Richland, WA 99352 (United States); Waters, Katrina M. [Superfund Research Center, Oregon State University, Corvallis, OR 97331 (United States); Biological Sciences Division, Pacific Northwest National Laboratory, Richland, WA 99352 (United States); Tilton, Susan C. [Department of Environmental and Molecular Toxicology, Oregon State University, Corvallis, OR 97331 (United States); Superfund Research Center, Oregon State University, Corvallis, OR 97331 (United States); Krueger, Sharon K. [Department of Environmental and Molecular Toxicology, Oregon State University, Corvallis, OR 97331 (United States); Superfund Research Center, Oregon State University, Corvallis, OR 97331 (United States); Linus Pauling Institute, Oregon State University, Corvallis, OR 97331 (United States); and others

    2015-09-01

    FVB/N mice wild-type, heterozygous or null for Cyp 1b1 were used in a two-stage skin tumor study comparing PAH, benzo[a]pyrene (BaP), dibenzo[def,p]chrysene (DBC), and coal tar extract (CTE, SRM 1597a). Following 20 weeks of promotion with TPA the Cyp 1b1 null mice, initiated with DBC, exhibited reductions in incidence, multiplicity, and progression. None of these effects were observed with BaP or CTE. The mechanism of Cyp 1b1-dependent alteration of DBC skin carcinogenesis was further investigated by determining expression of select genes in skin from DBC-treated mice 2, 4 and 8 h post-initiation. A significant reduction in levels of Cyp 1a1, Nqo1 at 8 h and Akr 1c14 mRNA was observed in Cyp 1b1 null (but not wt or het) mice, whereas no impact was observed in Gst a1, Nqo 1 at 2 and 4 h or Akr 1c19 at any time point. Cyp 1b1 mRNA was not elevated by DBC. The major covalent DNA adducts, dibenzo[def,p]chrysene-(±)-11,12-dihydrodiol-cis and trans-13,14-epoxide-deoxyadenosine (DBCDE-dA) were quantified by UHPLC-MS/MS 8 h post-initiation. Loss of Cyp1 b1 expression reduced DBCDE-dA adducts in the skin but not to a statistically significant degree. The ratio of cis- to trans-DBCDE-dA adducts was higher in the skin than other target tissues such as the spleen, lung and liver (oral dosing). These results document that Cyp 1b1 plays a significant role in bioactivation and carcinogenesis of DBC in a two-stage mouse skin tumor model and that loss of Cyp 1b1 has little impact on tumor response with BaP or CTE as initiators. - Highlights: • Cyp1b1 null mice exhibit lower skin cancer sensitivity to DBC but not BaP or CTE. • Cyp1b1 expression impacts expression of other PAH metabolizing enzymes. • cis/trans-DBCDE-dA ratio significantly higher in the skin than the spleen, lung or liver • Potency of DBC and CTE in mouse skin is higher than predicted by RPFs.

  6. Cytochrome P450 1b1 in polycyclic aromatic hydrocarbon (PAH)-induced skin carcinogenesis: Tumorigenicity of individual PAHs and coal-tar extract, DNA adduction and expression of select genes in the Cyp1b1 knockout mouse.

    Science.gov (United States)

    Siddens, Lisbeth K; Bunde, Kristi L; Harper, Tod A; McQuistan, Tammie J; Löhr, Christiane V; Bramer, Lisa M; Waters, Katrina M; Tilton, Susan C; Krueger, Sharon K; Williams, David E; Baird, William M

    2015-09-01

    FVB/N mice wild-type, heterozygous or null for Cyp 1b1 were used in a two-stage skin tumor study comparing PAH, benzo[a]pyrene (BaP), dibenzo[def,p]chrysene (DBC), and coal tar extract (CTE, SRM 1597a). Following 20 weeks of promotion with TPA the Cyp 1b1 null mice, initiated with DBC, exhibited reductions in incidence, multiplicity, and progression. None of these effects were observed with BaP or CTE. The mechanism of Cyp 1b1-dependent alteration of DBC skin carcinogenesis was further investigated by determining expression of select genes in skin from DBC-treated mice 2, 4 and 8h post-initiation. A significant reduction in levels of Cyp 1a1, Nqo1 at 8h and Akr 1c14 mRNA was observed in Cyp 1b1 null (but not wt or het) mice, whereas no impact was observed in Gst a1, Nqo 1 at 2 and 4h or Akr 1c19 at any time point. Cyp 1b1 mRNA was not elevated by DBC. The major covalent DNA adducts, dibenzo[def,p]chrysene-(±)-11,12-dihydrodiol-cis and trans-13,14-epoxide-deoxyadenosine (DBCDE-dA) were quantified by UHPLC-MS/MS 8h post-initiation. Loss of Cyp1 b1 expression reduced DBCDE-dA adducts in the skin but not to a statistically significant degree. The ratio of cis- to trans-DBCDE-dA adducts was higher in the skin than other target tissues such as the spleen, lung and liver (oral dosing). These results document that Cyp 1b1 plays a significant role in bioactivation and carcinogenesis of DBC in a two-stage mouse skin tumor model and that loss of Cyp 1b1 has little impact on tumor response with BaP or CTE as initiators. PMID:26049101

  7. Massive consumption of gelatinous plankton by Mediterranean apex predators.

    Directory of Open Access Journals (Sweden)

    Luis Cardona

    Full Text Available Stable isotopes of carbon and nitrogen were used to test the hypothesis that stomach content analysis has systematically overlooked the consumption of gelatinous zooplankton by pelagic mesopredators and apex predators. The results strongly supported a major role of gelatinous plankton in the diet of bluefin tuna (Thunnus thynnus, little tunny (Euthynnus alletteratus, spearfish (Tetrapturus belone and swordfish (Xiphias gladius. Loggerhead sea turtles (Caretta caretta in the oceanic stage and ocean sunfish (Mola mola also primarily relied on gelatinous zooplankton. In contrast, stable isotope ratios ruled out any relevant consumption of gelatinous plankton by bluefish (Pomatomus saltatrix, blue shark (Prionace glauca, leerfish (Lichia amia, bonito (Sarda sarda, striped dolphin (Stenella caerueloalba and loggerhead sea turtles (Caretta caretta in the neritic stage, all of which primarily relied on fish and squid. Fin whales (Balaenoptera physalus were confirmed as crustacean consumers. The ratios of stable isotopes in albacore (Thunnus alalunga, amberjack (Seriola dumerili, blue butterfish (Stromaeus fiatola, bullet tuna (Auxis rochei, dolphinfish (Coryphaena hyppurus, horse mackerel (Trachurus trachurus, mackerel (Scomber scombrus and pompano (Trachinotus ovatus were consistent with mixed diets revealed by stomach content analysis, including nekton and crustaceans, but the consumption of gelatinous plankton could not be ruled out completely. In conclusion, the jellyvorous guild in the Mediterranean integrates two specialists (ocean sunfish and loggerhead sea turtles in the oceanic stage and several opportunists (bluefin tuna, little tunny, spearfish, swordfish and, perhaps, blue butterfish, most of them with shrinking populations due to overfishing.

  8. Galaxy cluster scaling relations measured with APEX-SZ

    Science.gov (United States)

    Bender, A. N.; Kennedy, J.; Ade, P. A. R.; Basu, K.; Bertoldi, F.; Burkutean, S.; Clarke, J.; Dahlin, D.; Dobbs, M.; Ferrusca, D.; Flanigan, D.; Halverson, N. W.; Holzapfel, W. L.; Horellou, C.; Johnson, B. R.; Kermish, Z. D.; Klein, M.; Kneissl, R.; Lanting, T.; Lee, A. T.; Mehl, J.; Menten, K. M.; Muders, D.; Nagarajan, A.; Pacaud, F.; Reichardt, C. L.; Richards, P. L.; Schaaf, R.; Schwan, D.; Sommer, M. W.; Spieler, H.; Tucker, C.; Westbrook, B.

    2016-08-01

    We present thermal Sunyaev-Zel'dovich effect (SZE) measurements for 42 galaxy clusters observed at 150 GHz with the APEX-SZ experiment. For each cluster, we model the pressure profile and calculate the integrated Comptonization Y to estimate the total thermal energy of the intracluster medium (ICM). We compare the measured Y values to X-ray observables of the ICM from the literature (cluster gas mass Mgas, temperature TX, and YX = MgasTX) that relate to total cluster mass. We measure power-law scaling relations, including an intrinsic scatter, between the SZE and X-ray observables for three subsamples within the set of 42 clusters that have uniform X-ray analysis in the literature. We observe that differences between these X-ray analyses introduce significant variance into the measured scaling relations, particularly affecting the normalization. For all three subsamples, we find results consistent with a self-similar model of cluster evolution dominated by gravitational effects. Comparing to predictions from numerical simulations, these scaling relations prefer models that include cooling and feedback in the ICM. Lastly, we measure an intrinsic scatter of ˜28 per cent in the Y - YX scaling relation for all three subsamples.

  9. Massive consumption of gelatinous plankton by Mediterranean apex predators.

    Science.gov (United States)

    Cardona, Luis; Álvarez de Quevedo, Irene; Borrell, Assumpció; Aguilar, Alex

    2012-01-01

    Stable isotopes of carbon and nitrogen were used to test the hypothesis that stomach content analysis has systematically overlooked the consumption of gelatinous zooplankton by pelagic mesopredators and apex predators. The results strongly supported a major role of gelatinous plankton in the diet of bluefin tuna (Thunnus thynnus), little tunny (Euthynnus alletteratus), spearfish (Tetrapturus belone) and swordfish (Xiphias gladius). Loggerhead sea turtles (Caretta caretta) in the oceanic stage and ocean sunfish (Mola mola) also primarily relied on gelatinous zooplankton. In contrast, stable isotope ratios ruled out any relevant consumption of gelatinous plankton by bluefish (Pomatomus saltatrix), blue shark (Prionace glauca), leerfish (Lichia amia), bonito (Sarda sarda), striped dolphin (Stenella caerueloalba) and loggerhead sea turtles (Caretta caretta) in the neritic stage, all of which primarily relied on fish and squid. Fin whales (Balaenoptera physalus) were confirmed as crustacean consumers. The ratios of stable isotopes in albacore (Thunnus alalunga), amberjack (Seriola dumerili), blue butterfish (Stromaeus fiatola), bullet tuna (Auxis rochei), dolphinfish (Coryphaena hyppurus), horse mackerel (Trachurus trachurus), mackerel (Scomber scombrus) and pompano (Trachinotus ovatus) were consistent with mixed diets revealed by stomach content analysis, including nekton and crustaceans, but the consumption of gelatinous plankton could not be ruled out completely. In conclusion, the jellyvorous guild in the Mediterranean integrates two specialists (ocean sunfish and loggerhead sea turtles in the oceanic stage) and several opportunists (bluefin tuna, little tunny, spearfish, swordfish and, perhaps, blue butterfish), most of them with shrinking populations due to overfishing. PMID:22470416

  10. Expression of calcium/calAulin kinaseⅡα in fragile X mental retardation 1 gene knockout brain tissues in mice%CaMKⅡα在FMR1基因敲除小鼠脑组织中的表达

    Institute of Scientific and Technical Information of China (English)

    韦朝霞; 陈盛强; 陈希; 戴丽军

    2013-01-01

    目的 观察FMRI基因敲除型(KO)小鼠脑组织中钙/钙调素依赖蛋白激酶Ⅱα(CaMKⅡα)表达的改变,探讨CaMKⅡα是否为脆性X综合征相关蛋白(FMRP)的下调蛋白. 方法 PCR鉴定FVB近交系小鼠的基因型,按基因型的不同分为KO组和野生型(WT)组,每组10只.免疫组化染色检测KO及WT小鼠脑组织CaMKⅡα的表达与分布,用图像分析仪分别采集不同脑区免疫信号的吸光度(A)值进行比较. 结果 免疫组化染色检测显示KO与WT小鼠各个脑区普遍存在阳性信号;神经元胞浆尤其是靠近胞体的近端突起上信号呈强阳性,树突中亦有阳性信号,轴突上信号较弱;KO小鼠各脑区CaMKⅡα阳性信号的A值均较WT小鼠显著增高,差异有统计学意义(P<0.05). 结论 CaMKⅡα在成年KO小鼠各脑区的表达均显著增多,提示FMRP负性调节CaMKⅡα的表达.%Objective To observe the expression of calcium/calAulin kinaseⅡα (CaMKⅡα) in the brain tissues of fragile X mental retardation 1 (FMR1) gene knockout (KO) mice to investigate whether CaMKⅡα is regulated by fragile X mental retardation protein (FMRP).Methods According to the gene types of the FVB inbred mice identified by PCR,20 mice were divided into KO group and WT (wide type) group (n=10).The subcellular distribution and expression of CaMKⅡα were observed by immunohistochemical staining; the mean optical density (A) values of immunostaining signal of CaMKⅡα in various brain regions,including the motor cortex,temporal cortex,amygdala,hypothalamus and hippocampus,were determined by IBAS 2.0 image-analyzed system.Results CaMKⅡα immunoreactive cells were abundantly found in all brain subregions of KO and WT mice; especial positive signal was noted in the proximal processes of neurons,so as to those in the dendrite; week signcal was observed in the axon.No distributional difference was found between KO and WT mice.As compared with those in the WT mice,the A values were

  11. MicroRNA-134在FMR1基因敲除鼠脑组织中的表达及意义%Expression and significance of microRNA-134 in mouse brain tissue with FMR1 gene knockout

    Institute of Scientific and Technical Information of China (English)

    曾志涌; 邸伟; 肖都; 孙逊沙; 王玉良; 欧阳梅; 易咏红

    2010-01-01

    Objective To observe the expression ofmicroRNA-134 (miR-134) in the mouse brain tissue with FMR1 gene knockout during the different development periods and its expression characteristic, and explore whether the deficiency of fragile X mental retardation protein (FMRP) can induce the changes of miR-134 transcription. Methods FVB strain male mice, including FMR1 gene knockout (KO, n=15) and their wild type (WT, n=15) counterparts were chosen in the experiment. The expressions of miR-134 in the brain tissues of these KO mice that were 0 d, 4 and 6 w old and the age-matched WT mice were detected by qRT-PCR. Results The transcriptional level of miR-134 in the brain tissue of KO mice had no significant difference as compared with that of age-matched WT mice (P>0.05). The transcriptional levels of miR-134 in 6-w-old KO and WT mice were significantly decreased as compared with the newbom and 4-w-old same genotype mice (P<0.05). Conclusion The absence of FMRP does not influence the transcription of miR-134 and the transcriptional level of miR-134 in the brain tissues maintains a high level during the developmental stage of the nervous system and gradually decreases to a low level after grow-up, demonstrating its important role in regulating the development of nervous system.%目的 观察脆性X综合征(FXS)模型小鼠不同发育时期脑组织中microRNA-134(miR-134)的表达,明确miR-134的表达特点及脆性X智力低下蛋白(FMRP)缺失是否导致miR-134转录的改变. 方法 应用荧光实时定量PCR检测FVB近交系雄性0 d、4、6周(W)龄FMR1基因敲除型(KO)(KO0d、KO4w、KO6w)和同龄野生型(WT)(WT0d、WT4w、WT6w)小鼠脑组织中miR-134的表达(n=5). 结果同龄KO与WT小鼠miR-134的转录表达量差异无统计学意义(P>0.05);KO6w小鼠脑组织miR-134的转录表达量低于KO0d和KO2w小鼠,WT6w小鼠脑组织miR-134的转录表达量也低于WT0d和WT2w小鼠,差异均有统计学意义(P<0.05). 结论 FMRp

  12. Expressions of Drebrins and lcam-5 in mouse cerebral cortex with Fmr-1 gene knockout and their significance in fragile X syndrome%Drebrins和Icam-5在Fmr-1基因敲除鼠大脑皮层的表达和意义

    Institute of Scientific and Technical Information of China (English)

    徐琴; 竺智伟; 赵正言

    2012-01-01

    [Objective]To investigate and compare the changes of Drebrin A,Drebrin E and lcam-5 mRNA levels in the cerebral cortex of Frr-1 gene knockout mouse during brain development periods.[Methods]Fmr-1 gene knockout (KO) male mice and their wild type (WT) counterparts were chosen in our experiment (4≤n≤ 10);the levels of target mRNAs were detected by real time quantitative PCR;check points were set on the 7th,14th,21th and 28rh postnatal d.[Results] The mRNA level of Drebrin A in the KO group was significantly lower than that in the WT group on the 14th postnatal d,while that of Drebrin E was significantly higher than that in the WT group (P<0.05).The mRNA level of lcam-5 in the KO group was significantly higher than that in the WT group on the 14th and 21th postnatal d (P<0.05).[Conclusion] The delayed shift of Drebrin A to Drebrin E and transitional over-expression of lcam-5 in developmental cerebral cortex are the reasons for mental retardation in Fragile X Syndrome.%目的 观察脆性X综合征(FXS)模型小鼠不同发育时期大脑皮层中Drebrin A、Drebrin E及Icam-5 mRNA水平变化情况及意义.方法 应用荧光实时定量PCR(RT-PCR)法检测FmrJ基因敲除KO小鼠及野生健康对照小鼠H出生后第7天、第14天、第21天和第28天大脑皮层Drebrin A、Drebrin E及Icam-5 mRNA的表达(4≤n≤10).结果 KO组小鼠出生后第14天Drebrin A mRNA水平较健康对照组小鼠明显降低,而同时间Drebrin E mRNA水平较健康对照组小鼠明显增高,差异均有统计学意义(P<0.05);KO组小鼠Icam-5 mRNA水平在出生后第14和21天均明显高于健康对照组,差异均有统计学意义(P<0.05).结论 Drebrin A和Drebrin E在大脑皮层发育期的表达交替延迟及Icam-5的一过性过度表达是FXS智力低下的原因之一.

  13. Electronic apex locator: A comprehensive literature review — Part II: Effect of different clinical and technical conditions on electronic apex locator′s accuracy

    Directory of Open Access Journals (Sweden)

    Hamid Razavian

    2014-01-01

    Full Text Available Introduction: To investigate the effects of different clinical and technical conditions on the accuracy of electronic apex locators (EALs. Materials and Methods: "Tooth apex," "dental instrument," "odontometry," "electronic medical," and "electronic apex locator" were searched as primary identifiers via Medline/PubMed, Cochrane library, and Scopus data base up to 30 July 2013. Original articles that fulfilled the inclusion criteria were selected and reviewed. Results: Out of 402 relevant studies, 183 were selected based on the inclusion criteria. In this part, 75 studies are presented. Pulp vitality conditions and root resorption, types of files and irrigating materials do not affect an EAL′s accuracy; however, the file size and foramen diameter can affect its accuracy. Conclusions: Various clinical conditions such as the file size and foramen diameter may affect EALs′ accuracy. However, more randomized clinical trials are needed for definitive conclusion.

  14. The vasopressin deficient Brattleboro rats : A natural knockout model used in the search for CNS effects of vasopressin

    NARCIS (Netherlands)

    Bohus, B; de Wied, D

    1998-01-01

    Behavioral neuroscience is using mon and more gene knockout techniques to produce animals with a specific deletion. These studies have their precedent in nature. A mutation may result in a limited genetic defect, as seen in the vasopressin (VP) deficiency in the Brattleboro rat. The mutation is in a

  15. Transient in utero knockout (TIUKO of C-MYC affects late lung and intestinal development in the mouse

    Directory of Open Access Journals (Sweden)

    Zhou Pengbo

    2004-04-01

    Full Text Available Abstract Background Developmentally important genes often result in early lethality in knockout animals. Thus, the direct role of genes in late gestation organogenesis cannot be assessed directly. In utero delivery of transgenes was shown previously to result in high efficiency transfer to pulmonary and intestinal epithelial stem cells. Thus, this technology can be used to evaluate late gestation development. Results In utero gene transfer was used to transfer adenovirus with either an antisense c-myc or a C-MYC ubiquitin targeting protein to knockout out c-myc expression in late gestation lung and intestines. Using either antisense or ubiquitin mediated knockout of C-MYC levels in late gestation resulted in similar effects. Decreased complexity was observed in both intestines and lungs. Stunted growth of villi was evident in the intestines. In the lung, hypoplastic lungs with disrupted aveolarization were observed. Conclusions These data demonstrated that C-MYC was required for cell expansion and complexity in late gestation lung and intestinal development. In addition they demonstrate that transient in utero knockout of proteins may be used to determine the role of developmentally important genes in the lungs and intestines.

  16. Generation and characterisation of keratin 7 (K7 knockout mice.

    Directory of Open Access Journals (Sweden)

    Aileen Sandilands

    Full Text Available Keratin 7 (K7 is a Type II member of the keratin superfamily and despite its widespread expression in different types of simple and transitional epithelia, its functional role in vivo remains elusive, in part due to the lack of any appropriate mouse models or any human diseases that are associated with KRT7 gene mutations. Using conventional gene targeting in mouse embryonic stem cells, we report here the generation and characterisation of the first K7 knockout mouse. Loss of K7 led to increased proliferation of the bladder urothelium although this was not associated with hyperplasia. K18, a presumptive type I assembly partner for K7, showed reduced expression in the bladder whereas K20, a marker of the terminally differentiated superficial urothelial cells was transcriptionally up-regulated. No other epithelia were seen to be adversely affected by the loss of K7 and western blot and immunofluorescence microscopy analysis revealed that the expression of K8, K18, K19 and K20 were not altered in the absence of K7, with the exception of the kidney where there was reduced K18 expression.

  17. Agrobacterium-mediated transformation of finger millet (Eleusine coracana (L.) Gaertn.) using shoot apex explants.

    Science.gov (United States)

    Ceasar, S Antony; Ignacimuthu, S

    2011-09-01

    A new Agrobacterium-mediated transformation system was developed for finger millet using shoot apex explants. The Agrobacterium strain LBA4404 harboring binary vector pCAMBIA1301, which contained hygromycin phosphotransferase (hptII) as selectable marker gene and β-glucuronidase (GUS) as reporter gene, was used for optimization of transformation conditions. Two finger millet genotypes, GPU 45 and CO 14, were used in this study. The optimal conditions for the Agrobacterium-mediated transformation of finger millet were found to be the co-cultivation of explants obtained on the 16th day after callus induction (DACI), exposure of explants for 30 min to agrobacterial inoculum and 3 days of co-cultivation on filter paper placed on medium supplemented with 100 μM acetosyringone (AS). Addition of 100 μM L: -cysteine in the selection medium enhanced the frequency of transformation and transgenic plant recovery. Both finger millet genotypes were transformed by Agrobacterium. A frequency of 19% transient expression with 3.8% stable transformation was achieved in genotype GPU 45 using optimal conditions. Five stably transformed plants were fully characterized by Southern blot analysis. A segregation analysis was also performed in four R(1) progenies, which showed normal Mendelian pattern of transgene segregation. The inheritance of transgenes in R(1) progenies was also confirmed by Southern blot analysis. This is the first report on Agrobacterium-mediated transformation of finger millet. This study underpins the introduction of numerous agronomically important genes into the genome of finger millet in the future.

  18. Rapid knockout and reporter mouse line generation and breeding colony establishment using EUCOMM conditional-ready embryonic stem cells: A case study

    Directory of Open Access Journals (Sweden)

    James L. J. Coleman

    2015-06-01

    Full Text Available As little as a decade ago, generation of a single knockout mouse line was an expensive and time-consuming undertaking available to relatively few researchers. The International Knockout Mouse Consortium, established in 2007, has revolutionized the use of such models by creating an open-access repository of ES cells that, through sequential breeding with first FlpE and then Cre recombinase transgenic mice, facilitates germline global or conditional deletion of almost every gene in the mouse genome. In this Case Study, we describe our experience using the repository to create mouse lines for a variety of experimental purposes. Specifically, we discuss the process of obtaining germline transmission of two EUCOMM ‘knockout-first’ gene targeted constructs and the advantages and pitfalls of using this system. We then outline our breeding strategy and the outcomes of our efforts to generate global and conditional knockouts and reporter mice for the genes of interest. Line maintenance, removal of recombinase transgenes and cryopreservation are also considered. Our approach led to the generation of heterozygous knockout mice within 6 months of commencing breeding to the founder mice. By describing our experiences with the EUCOMM ES cells and subsequent breeding steps, we hope to assist other researchers with the application of this valuable approach to generating versatile knockout mouse lines.

  19. Apex determination and detection of stellar clumps in the open cluster M 67

    CERN Document Server

    Vereshchagin, S V; Sariya, Devesh P; Yadav, R K S; Kumar, Brijesh

    2014-01-01

    We determined the cluster apex coordinates, studied the substructures and performed membership analysis in the central part (34'X33') of the open cluster M 67. We used the individual stellar apexes method developed earlier and classical technique of proper motion diagrams in coordinate system connected with apex. The neighbour-to-neighbour distance technique was applied to detect space details. The membership list was corrected and some stars were excluded from the most probable members list. The apex coordinates have been determined as: A0=132.97deg+/-0.81deg and D0=11.85deg+/-0.90deg. The 2D-space star density field was analysed and high degree of inhomogeneity was found.

  20. Frequency of the external resorptions of root apex

    Directory of Open Access Journals (Sweden)

    Opačić-Galić Vanja

    2004-01-01

    Full Text Available Root resorptions present a significant problem in endodontic therapy of the affected teeth and in dentistry in general. The objective of this study was to analyze, based on epidemiological and statistical research, the frequency of clinical incidence of pathological root resorptions in everyday practice related to localization, type of tooth, age and sex of patients. Radiographie documentation of patients treated from 1997 till 2002 at the Department of Conservative Dentistry and Endodontics, Faculty of Stomatology in Belgrade, was used as baseline for this study. Retroalveolar radiographs of teeth with visible signs of resorptions were singled out from 15654 patients' clinical records used for this study. The external resorptions were shown as radiolucent areas localized on various outer root surfaces, followed by significant or less significant resorption of lamina dura and alveolar bone. Out of all teeth analyzed in this study, 594 (3.79% showed some kind of resorption. The external resorptions were found to be more present in the upper jaw (55.10% and molars (50.30% than in the lower jaw (44.90% and single root teeth (49.70%, but in both cases without significant statistical differences. The most frequent localization of resorptions was root apex (82.44%. In regard to age, the most frequent resorptions were recorded in patients aged between 21 and 30 years (28.40%, and the lowest incidence was found in the youngest population (5.51%. The results also showed that resorptions were more frequent among the female population (59.04% than among the male population (40.96%. Based on these results, we may conclude that the external root resorptions are not a frequent clinical phenomenon. Proper and early diagnostics of such tissue pathology is one of the basic prerequisites for successful endodontic therapy of the affected root.

  1. Crossing latitudes--long-distance tracking of an apex predator.

    Directory of Open Access Journals (Sweden)

    Luciana C Ferreira

    Full Text Available Tiger sharks (Galeocerdo cuvier are apex predators occurring in most tropical and warm temperate marine ecosystems, but we know relatively little of their patterns of residency and movement over large spatial and temporal scales. We deployed satellite tags on eleven tiger sharks off the north-western coast of Western Australia and used the Brownian Bridge kernel method to calculate home ranges and analyse movement behaviour. One individual recorded one of the largest geographical ranges of movement ever reported for the species, travelling over 4000 km during 517 days of monitoring. Tags on the remainder of the sharks reported for shorter periods (7-191 days. Most of these sharks had restricted movements and long-term (30-188 days residency in coastal waters in the vicinity of the area where they were tagged. Core home range areas of sharks varied greatly from 1166.9 to 634,944 km2. Tiger sharks spent most of their time in water temperatures between 23°-26°C but experienced temperatures ranging from 6°C to 33°C. One shark displayed seasonal movements among three distinct home range cores spread along most of the coast of Western Australia and generalized linear models showed that this individual had different patterns of temperature and depth occupancy in each region of the coast, with the highest probability of residency occurring in the shallowest areas of the coast with water temperatures above 23°C. These results suggest that tiger sharks can migrate over very large distances and across latitudes ranging from tropical to the cool temperate waters. Such extensive long-term movements may be a key element influencing the connectivity of populations within and among ocean basins.

  2. Forest species mapping using airborne hyperspectral APEX data

    Directory of Open Access Journals (Sweden)

    Tagliabue Giulia

    2016-03-01

    Full Text Available The accurate mapping of forest species is a very important task in relation to the increasing need to better understand the role of the forest ecosystem within environmental dynamics. The objective of this paper is the investigation of the potential of a multi-temporal hyperspectral dataset for the production of a thematic map of the dominant species in the Forêt de Hardt (France. Hyperspectral data were collected in June and September 2013 using the Airborne Prism EXperiment (APEX sensor, covering the visible, near-infrared and shortwave infrared spectral regions with a spatial resolution of 3 m by 3 m. The map was realized by means of a maximum likelihood supervised classification. The classification was first performed separately on images from June and September and then on the two images together. Class discrimination was performed using as input 3 spectral indices computed as ratios between red edge bands and a blue band for each image. The map was validated using a testing set selected on the basis of a random stratified sampling scheme. Results showed that the algorithm performances improved from an overall accuracy of 59.5% and 48% (for the June and September images, respectively to an overall accuracy of 74.4%, with the producer’s accuracy ranging from 60% to 86% and user’s accuracy ranging from 61% to 90%, when both images (June and September were combined. This study demonstrates that the use of multi-temporal high-resolution images acquired in two different vegetation development stages (i.e., 17 June 2013 and 4 September 2013 allows accurate (overall accuracy 74.4% local-scale thematic products to be obtained in an operational way.

  3. Noise alters hair-bundle mechanics at the cochlear apex

    Science.gov (United States)

    Strimbu, C. Elliott; Fridberger, Anders

    2015-12-01

    Exposure to loud sounds can lead to both permanent and short term changes in auditory sensitivity. Permanent hearing loss is often associated with gross changes in cochlear morphology including the loss of hair cells and auditory nerve fibers while the mechanisms of short term threshold shifts are much less well understood and may vary at different locations across the cochlea. Previous reports suggest that exposure to loud sounds leads to a decrease in the cochlear microphonic potential and in the stiffness of the organ of Corti. Because the cochlear microphonic reflects changes in the membrane potential of the hair cells, this suggests that hair-bundle motion should be reversibly altered following exposure to loud sounds. Using an in vitro preparation of the guinea pig temporal bone we investigate changes in the micro-mechanical response near the cochlear apex following a brief (up to 10 - 20 minutes) exposure to loud (˜ 120 dB) tones near the best frequency at this location. We use time-resolved confocal imaging to record the motion of outer hair cell bundles before and after acoustic overstimulation. We have also recorded larger-scale structural views of the organ of Corti before and after exposure to the loud sound. Conventional electrophysiological techniques are used measure the cochlear microphonic potential. As has been previously reported, following acoustic overexposure the cochlear microphonic declines in value and typically recovers on the order of 30 - 60 minutes. Hair-bundle trajectories are affected following the loud sound and typically recover on a somewhat faster time scale than the microphonic potential, although the results vary considerably across preparations. Preliminary results also suggest reversible changes in the hair cell's resting potential following the loud sound.

  4. Sixty-seven Cases of Abnormal Movement of the Cardiac Apex Treated with Bu Xin Tang

    Institute of Scientific and Technical Information of China (English)

    王平; 耿世钊

    2002-01-01

    @@ Bu Xin Tang (heart-reinforcement decoction) was used to treat 67 cases of abnormal movement of the cardiac apex based on differentiation of symptoms and signs. The results showed that, in most patients, there were remarkable improvement notonly for the symptoms but also for the abnormal movement of the cardiac apex. The cured plus remarkably effective rate was 87%, suggesting that it can postponeor prevent coronary heart attacks for the patient of prophase coronary heart disease.

  5. High-resolution NO₂ remote sensing from the Airborne Prism EXperiment (APEX) imaging spectrometer

    OpenAIRE

    Popp, C; Brunner, D.; Damm, A.; Van Roozendael, M.; Fayt, C.; Buchmann, B

    2012-01-01

    We present and evaluate the retrieval of high spatial resolution maps of NO2 vertical column densities (VCD) from the Airborne Prism EXperiment (APEX) imaging spectrometer. APEX is a novel instrument providing airborne measurements of unique spectral and spatial resolution and coverage as well as high signal stability. In this study, we use spectrometer data acquired over Zurich, Switzerland, in the morning and late afternoon during a flight campaign on a cloud-free summer d...

  6. SEPIA — A New Instrument for the Atacama Pathfinder Experiment (APEX) Telescope

    Science.gov (United States)

    Immer, K.; Belitsky, V.; Olberg, M.; De Breuck, C.; Conway, J.; Montenegro-Montes, F. M.; Perez-Beaupuits, J.-P.; Torstensson, K.; Billade, B.; De Beck, E.; Ermakov, A.; Ferm, S.-E.; Fredrixon, M.; Lapkin, I.; Meledin, D.; Pavolotsky, A.; Strandberg, M.; Sundin, E.; Arumugam, V.; Galametz, M.; Humphreys, E.; Klein, T.; Adema, J.; Barkhof, J.; Baryshev, A.; Boland, W.; Hesper, R.; Klapwijk, T. M.

    2016-09-01

    The Swedish-ESO PI receiver for APEX (SEPIA) was installed at the APEX telescope in 2015. This instrument currently contains ALMA Band 5 (157-212 GHz) and Band 9 (600-722 GHz) receivers. Commissioning and science verification for Band 5 have been successfully completed but are still ongoing for Band 9. The SEPIA instrument is briefly described and the commissioning of the Band 5 receiver and results from the first science observations are presented.

  7. APEX 3: a multi-purpose test platform for auditory psychophysical experiments.

    Science.gov (United States)

    Francart, Tom; van Wieringen, Astrid; Wouters, Jan

    2008-07-30

    APEX 3 is a software test platform for auditory behavioral experiments. It provides a generic means of setting up experiments without any programming. The supported output devices include sound cards and cochlear implants from Cochlear Corporation and Advanced Bionics Corporation. Many psychophysical procedures are provided and there is an interface to add custom procedures. Plug-in interfaces are provided for data filters and external controllers. APEX 3 is supported under Linux and Windows and is available free of charge. PMID:18538414

  8. Evolution mediates the effects of apex predation on aquatic food webs.

    Science.gov (United States)

    Urban, Mark C

    2013-07-22

    Ecological and evolutionary mechanisms are increasingly thought to shape local community dynamics. Here, I evaluate if the local adaptation of a meso-predator to an apex predator alters local food webs. The marbled salamander (Ambystoma opacum) is an apex predator that consumes both the spotted salamander (Ambystoma maculatum) and shared zooplankton prey. Common garden experiments reveal that spotted salamander populations which co-occur with marbled salamanders forage more intensely than those that face other predator species. These foraging differences, in turn, alter the diversity, abundance and composition of zooplankton communities in common garden experiments and natural ponds. Locally adapted spotted salamanders exacerbate prey biomass declines associated with apex predation, but dampen the top-down effects of apex predation on prey diversity. Countergradient selection on foraging explains why locally adapted spotted salamanders exacerbate prey biomass declines. The two salamander species prefer different prey species, which explains why adapted spotted salamanders buffer changes in prey composition owing to apex predation. Results suggest that local adaptation can strongly mediate effects from apex predation on local food webs. Community ecologists might often need to consider the evolutionary history of populations to understand local diversity patterns, food web dynamics, resource gradients and their responses to disturbance.

  9. Lack of association between the APEX1 Asp148Glu polymorphism and sporadic amyotrophic lateral sclerosis.

    Science.gov (United States)

    Coppedè, Fabio; Lo Gerfo, Annalisa; Carlesi, Cecilia; Piazza, Selina; Mancuso, Michelangelo; Pasquali, Livia; Murri, Luigi; Migliore, Lucia; Siciliano, Gabriele

    2010-02-01

    Impairments in DNA repair enzymes have been observed in amyotrophic lateral sclerosis (ALS) tissues, particularly in the activity of the apurinic/apyrimidinic endonuclease 1 (APEX1). Moreover, it was suggested that the common APEX1 Asp148Glu polymorphism might be associated with ALS risk. To further address this question we performed the present study aimed at evaluating the contribution of the APEX1 Asp148Glu polymorphism in sporadic ALS (sALS) risk and clinical presentation, including age and site of onset and disease progression. We screened 134 sALS Italian patients and 129 matched controls for the presence of the APEX1 Asp148Glu polymorphism. No difference in APEX1 Asp148Glu allele and genotype frequencies was found between the groups, nor was the polymorphism associated with age and site of onset or disease progression. Present results do not support a role for the APEX1 Asp148Glu polymorphism in sALS pathogenesis in the Italian population. PMID:18482781

  10. Cytoprotective role of autophagy against BH3 mimetic gossypol in ATG5 knockout cells generated by CRISPR-Cas9 endonuclease.

    Science.gov (United States)

    Kim, Na-Yeon; Han, Byeal-I; Lee, Michael

    2016-01-01

    Previously, we demonstrated the association between autophagy and gossypol-induced growth inhibition of mutant BRAF melanoma cells. Here, we investigate the role of autophagy in ATG5 knockout cell lines generated by the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas-mediated genome editing. The MTT assay revealed that the inhibitory effect of gossypol was weaker on ATG5 knockout cells than that on the wild type (WT) cells. The conversion of non-autophagic LC3-I to autophagic LC3-II and RT-PCR confirmed the functional gene knockout. However, Cyto-ID autophagy assay revealed that gossypol induced ATG5- and LC3-independent autophagy in ATG5 knockout cells. Moreover, gossypol acts as an autophagy inducer in ATG5 knockout cells while blocking the later stages of the autophagy process in WT cells, which was determined by measuring autophagic flux after co-treatment of gossypol with chloroquine (late-stage autophagy inhibitor). On the other hand, inhibition of autophagy with 3-MA or Beclin-1 siRNA caused a partial increase in the sensitivity to gossypol in ATG5 knockout cells, but not in the WT cells. Together, our findings suggest that the resistance to gossypol in ATG5 knockout cells is associated with increased cytoprotective autophagy, independent of ATG5.

  11. UFO in the Arabidopsis inflorescence apex is required for floral-meristem identity and bract suppression.

    Science.gov (United States)

    Hepworth, Shelley R; Klenz, Jennifer E; Haughn, George W

    2006-03-01

    The UNUSUAL FLORAL ORGANS (UFO) gene of Arabidopsis encodes an F-box protein required for the determination of floral-organ and floral-meristem identity. Mutation of UFO leads to dramatic changes in floral-organ type which are well-characterized whereas inflorescence defects are more subtle and less understood. These defects include an increase in the number of secondary inflorescences, nodes that alternate between forming flowers and secondary inflorescences, and nodes in which a single flower is subtended by a bract. Here, we show how inflorescence defects correlate with the abnormal development of floral primordia and establish a temporal requirement for UFO in this process. At the inflorescence apex of ufo mutants, newly formed primordia are initially bract-like. Expression of the floral-meristem identity genes LFY and AP1 are confined to a relatively small adaxial region of these primordia with expression of the bract-identity marker FIL observed in cells that comprise the balance of the primordia. Proliferation of cells in the adaxial region of these early primordia is delayed by several nodes such that primordia appear "chimeric" at several nodes, having visible floral and bract components. However, by late stage 2 of floral development, growth of the bract generally ceases and is overtaken by development of the floral primordium. This abnormal pattern of floral meristem development is not rescued by expression of UFO from the AP1 promoter, indicating that UFO is required prior to AP1 activation for normal development of floral primordia. We propose that UFO and LFY are jointly required in the inflorescence meristem to both promote floral meristem development and inhibit, in a non-cell autonomous manner, growth of the bract. PMID:16244866

  12. UFO in the Arabidopsis inflorescence apex is required for floral-meristem identity and bract suppression.

    Science.gov (United States)

    Hepworth, Shelley R; Klenz, Jennifer E; Haughn, George W

    2006-03-01

    The UNUSUAL FLORAL ORGANS (UFO) gene of Arabidopsis encodes an F-box protein required for the determination of floral-organ and floral-meristem identity. Mutation of UFO leads to dramatic changes in floral-organ type which are well-characterized whereas inflorescence defects are more subtle and less understood. These defects include an increase in the number of secondary inflorescences, nodes that alternate between forming flowers and secondary inflorescences, and nodes in which a single flower is subtended by a bract. Here, we show how inflorescence defects correlate with the abnormal development of floral primordia and establish a temporal requirement for UFO in this process. At the inflorescence apex of ufo mutants, newly formed primordia are initially bract-like. Expression of the floral-meristem identity genes LFY and AP1 are confined to a relatively small adaxial region of these primordia with expression of the bract-identity marker FIL observed in cells that comprise the balance of the primordia. Proliferation of cells in the adaxial region of these early primordia is delayed by several nodes such that primordia appear "chimeric" at several nodes, having visible floral and bract components. However, by late stage 2 of floral development, growth of the bract generally ceases and is overtaken by development of the floral primordium. This abnormal pattern of floral meristem development is not rescued by expression of UFO from the AP1 promoter, indicating that UFO is required prior to AP1 activation for normal development of floral primordia. We propose that UFO and LFY are jointly required in the inflorescence meristem to both promote floral meristem development and inhibit, in a non-cell autonomous manner, growth of the bract.

  13. The APEX Quantitative Proteomics Tool: Generating protein quantitation estimates from LC-MS/MS proteomics results

    Directory of Open Access Journals (Sweden)

    Saeed Alexander I

    2008-12-01

    Full Text Available Abstract Background Mass spectrometry (MS based label-free protein quantitation has mainly focused on analysis of ion peak heights and peptide spectral counts. Most analyses of tandem mass spectrometry (MS/MS data begin with an enzymatic digestion of a complex protein mixture to generate smaller peptides that can be separated and identified by an MS/MS instrument. Peptide spectral counting techniques attempt to quantify protein abundance by counting the number of detected tryptic peptides and their corresponding MS spectra. However, spectral counting is confounded by the fact that peptide physicochemical properties severely affect MS detection resulting in each peptide having a different detection probability. Lu et al. (2007 described a modified spectral counting technique, Absolute Protein Expression (APEX, which improves on basic spectral counting methods by including a correction factor for each protein (called Oi value that accounts for variable peptide detection by MS techniques. The technique uses machine learning classification to derive peptide detection probabilities that are used to predict the number of tryptic peptides expected to be detected for one molecule of a particular protein (Oi. This predicted spectral count is compared to the protein's observed MS total spectral count during APEX computation of protein abundances. Results The APEX Quantitative Proteomics Tool, introduced here, is a free open source Java application that supports the APEX protein quantitation technique. The APEX tool uses data from standard tandem mass spectrometry proteomics experiments and provides computational support for APEX protein abundance quantitation through a set of graphical user interfaces that partition thparameter controls for the various processing tasks. The tool also provides a Z-score analysis for identification of significant differential protein expression, a utility to assess APEX classifier performance via cross validation, and a

  14. Single and double metallothionein knockout in the nematode C. elegans reveals cadmium dependent and independent toxic effects on life history traits

    Energy Technology Data Exchange (ETDEWEB)

    Hughes, Sam [School of Biosciences, Cardiff University, Main Building, Park Place, Cardiff CF10 3TL (United Kingdom); School of Biomedical and Health Sciences, Pharmaceutical Sciences Research Division, King' s College London, 150 Stamford Street, London SE1 9NH (United Kingdom); Stuerzenbaum, Stephen R. [School of Biosciences, Cardiff University, Main Building, Park Place, Cardiff CF10 3TL (United Kingdom) and School of Biomedical and Health Sciences, Pharmaceutical Sciences Research Division, King' s College London, 150 Stamford Street, London SE1 9NH (United Kingdom)]. E-mail: stephen.sturzenbaum@kcl.ac.uk

    2007-01-15

    The genome of the nematode Caenorhabditis elegans contains two metallothionein genes, both involved in metal homeostasis and/or detoxification. Single metallothionein knockout mutants have been created and now, for the first time, a double mutant has been isolated. Life history studies in the presence or absence of cadmium showed that all metallothionein mutants are viable. Although cadmium did not influence longevity, a dose dependent reduction in total brood size and volumetric growth was observed in wild type animals, which was magnified in single knockouts and further exacerbated in the double knockout. However, the metallothionein deletion caused two effects that are independent of cadmium exposure, namely all knockout strains displayed a reduced total brood size and the deletion of both metallothionein loci caused a significant reduction in volumetric growth. In summary, metallothionein is undoubtedly an important player in cadmium detoxification, but evidently also an important factor in cadmium independent pathways. - Metallothionein is a modifier of life-history parameters.

  15. Bone growth and turnover in progesterone receptor knockout mice.

    Energy Technology Data Exchange (ETDEWEB)

    Rickard, David J.; Iwaniec, Urszula T.; Evans, Glenda; Hefferan, Theresa E.; Hunter, Jaime C.; Waters, Katrina M.; Lydon, John P.; O' Malley, Bert W.; Khosla, Sundeep; Spelsberg, Thomas C.; Turner, Russell T.

    2008-05-01

    The role of progesterone receptor (PR) signaling in skeletal metabolism is controversial. To address whether signaling through the PR is necessary for normal bone growth and turnover, we performed histomorphometric and mCT analyses of bone from homozygous female PR knockout (PRKO) mice at 6, 12, and 26 weeks of age. These mice possess a null mutation of the PR locus, which blocks the gene expression of A and B isoforms of PR. Body weight gain, uterine weight gain and tibia longitudinal bone growth was normal in PRKO mice. In contrast, total and cortical bone mass were increased in long bones of post-pubertal (12 and 26-week-old) PRKO mice, whereas cancellous bone mass was normal in the tibia but increased in the humerus. The striking 57% decrease in cancellous bone from the proximal tibia metaphysis which occurred between 6 and 26 weeks in WT mice was abolished in PRKO mice. The improved bone balance in aging PRKO mice was associated with elevated bone formation and a tendency toward reduced osteoclast perimeter. Taken together, these findings suggest that PR signaling in mice attenuates the accumulation of cortical bone mass during adolescence and is required for early age-related loss of cancellous bone.

  16. Generation and behavior characterization of CaMKIIβ knockout mice.

    Directory of Open Access Journals (Sweden)

    Adam D Bachstetter

    Full Text Available The calcium/calmodulin-dependent protein kinase II (CaMKII is abundant in the brain, where it makes important contributions to synaptic organization and homeostasis, including playing an essential role in synaptic plasticity and memory. Four genes encode isoforms of CaMKII (α, β, δ, γ, with CaMKIIα and CaMKIIβ highly expressed in the brain. Decades of molecular and cellular research, as well as the use of a large number of CaMKIIα mutant mouse lines, have provided insight into the pivotal roles of CaMKIIα in brain plasticity and cognition. However, less is known about the CaMKIIβ isoform. We report the development and extensive behavioral and phenotypic characterization of a CaMKIIβ knockout (KO mouse. The CaMKIIβ KO mouse was found to be smaller at weaning, with an altered body mass composition. The CaMKIIβ KO mouse showed ataxia, impaired forelimb grip strength, and deficits in the rotorod, balance beam and running wheel tasks. Interestingly, the CaMKIIβ KO mouse exhibited reduced anxiety in the elevated plus maze and open field tests. The CaMKIIβ KO mouse also showed cognitive impairment in the novel object recognition task. Our results provide a comprehensive behavioral characterization of mice deficient in the β isoform of CaMKII. The neurologic phenotypes and the construction of the genotype suggest the utility of this KO mouse strain for future studies of CaMKIIβ in brain structure, function and development.

  17. Sleep in Kcna2 knockout mice

    Directory of Open Access Journals (Sweden)

    Messing Albee

    2007-10-01

    Full Text Available Abstract Background Shaker codes for a Drosophila voltage-dependent potassium channel. Flies carrying Shaker null or hypomorphic mutations sleep 3–4 h/day instead of 8–14 h/day as their wild-type siblings do. Shaker-like channels are conserved across species but it is unknown whether they affect sleep in mammals. To address this issue, we studied sleep in Kcna2 knockout (KO mice. Kcna2 codes for Kv1.2, the alpha subunit of a Shaker-like voltage-dependent potassium channel with high expression in the mammalian thalamocortical system. Results Continuous (24 h electroencephalograph (EEG, electromyogram (EMG, and video recordings were used to measure sleep and waking in Kcna2 KO, heterozygous (HZ and wild-type (WT pups (P17 and HZ and WT adult mice (P67. Sleep stages were scored visually based on 4-s epochs. EEG power spectra (0–20 Hz were calculated on consecutive 4-s epochs. KO pups die by P28 due to generalized seizures. At P17 seizures are either absent or very rare in KO pups ( Conclusion Kv1.2, a mammalian homologue of Shaker, regulates neuronal excitability and affects NREM sleep.

  18. Gliosis after traumatic brain injury in conditional ephrinB2-knockout mice

    Institute of Scientific and Technical Information of China (English)

    LIU Ling; CHEN Xiao-lin; YANG Jian-kai; REN Ze-guang; WANG Shuo

    2012-01-01

    Background In response to the injury of the central nervous system (CNS),the astrocytes upregulate the expression of glial fibrillary acidic protein (GFAP),which largely contributes to the reactive gliosis after brain injury.The regulatory mechanism of this process is still not clear.In this study,we aimed to compare the ephrin-B2 deficient mice with the wild type ones with regard to gliosis after traumatic brain injury.Methods We generated ephrin-B2 knockout mice specifically in CNS astrocytes.Twelve mice from this gene-knockout strain were randomly selected along with twelve mice from the wild type littermates.In both groups,a modified controlled cortical impact injury model was applied to create a closed traumatic brain injury.Twenty-eight days after the injury,Nissl staining and GFAP immunofluorescence staining were used to compare the brain atrophy and GFAP immunoreactivity between the two groups.All the data were analyzed by t-test for between-group comparison.Results We successfully set up the conditional ephrin-B2 knockout mice strain,which was confirmed by genotyping and ephrin-B2/GFAP double staining.These mice developed normally without apparent abnormality in general appearance.Twenty-eight days following brain injury,histopathology revealed by immunohistochemistry showed different degrees of cerebral injuries in both groups.Compared with wild-type group,the ephrin-B2 knockout group exhibited less brain atrophy ratio for the injured hemispheres (P=0.005) and hippocampus (P=0.027).Also the wild-type group demonstrated greater GFAP immunoreactivity increment within hippocampal regions (P=0.008).Conclusions The establishment of conditional ephrin-B2 knockout mice provides us with a new way to explore the role of ephrin-B2 in astrocytes.Our findings revealed less atrophy and GFAP immunoreactivity in the knockout mice strain after traumatic brain injury,which implied ephrin-B2 could be one of the promoters to upregulate gliosis following brain injury.

  19. Characterization of Heterogeneous Prostate Tumors in Targeted Pten Knockout Mice.

    Directory of Open Access Journals (Sweden)

    Hanneke Korsten

    Full Text Available Previously, we generated a preclinical mouse prostate tumor model based on PSA-Cre driven inactivation of Pten. In this model homogeneous hyperplastic prostates (4-5m developed at older age (>10m into tumors. Here, we describe the molecular and histological characterization of the tumors in order to better understand the processes that are associated with prostate tumorigenesis in this targeted mouse Pten knockout model. The morphologies of the tumors that developed were very heterogeneous. Different histopathological growth patterns could be identified, including intraductal carcinoma (IDC, adenocarcinoma and undifferentiated carcinoma, all strongly positive for the epithelial cell marker Cytokeratin (CK, and carcinosarcomas, which were negative for CK. IDC pattern was already detected in prostates of 7-8 month old mice, indicating that it could be a precursor stage. At more than 10 months IDC and carcinosarcoma were most frequently observed. Gene expression profiling discriminated essentially two molecular subtypes, denoted tumor class 1 (TC1 and tumor class 2 (TC2. TC1 tumors were characterized by high expression of epithelial markers like Cytokeratin 8 and E-Cadherin whereas TC2 tumors showed high expression of mesenchyme/stroma markers such as Snail and Fibronectin. These molecular subtypes corresponded with histological growth patterns: where TC1 tumors mainly represented adenocarcinoma/intraductal carcinoma, in TC2 tumors carcinosarcoma was the dominant growth pattern. Further molecular characterization of the prostate tumors revealed an increased expression of genes associated with the inflammatory response. Moreover, functional markers for senescence, proliferation, angiogenesis and apoptosis were higher expressed in tumors compared to hyperplasia. The highest expression of proliferation and angiogenesis markers was detected in TC2 tumors. Our data clearly showed that in the genetically well-defined PSA-Cre;Pten-loxP/loxP prostate tumor

  20. Comparative morphology of dendritic arbors in populations of Purkinje cells in mouse sulcus and apex.

    Science.gov (United States)

    Nedelescu, Hermina; Abdelhack, Mohamed

    2013-01-01

    Foliation divides the mammalian cerebellum into structurally distinct subdivisions, including the concave sulcus and the convex apex. Purkinje cell (PC) dendritic morphology varies between subdivisions and changes significantly ontogenetically. Since dendritic morphology both enables and limits sensory-motor circuit function, it is important to understand how neuronal architectures differ between brain regions. This study employed quantitative confocal microcopy to reconstruct dendritic arbors of cerebellar PCs expressing green fluorescent protein and compared arbor morphology between PCs of sulcus and apex in young and old mice. Arbors were digitized from high z-resolution (0.25 µm) image stacks using an adaptation of Neurolucida's (MBF Bioscience) continuous contour tracing tool, designed for drawing neuronal somata. Reconstructed morphologies reveal that dendritic arbors of sulcus and apex exhibit profound differences. In sulcus, 72% of the young PC population possesses two primary dendrites, whereas in apex, only 28% do. Spatial constraints in the young sulcus cause significantly more dendritic arbor overlap than in young apex, a distinction that disappears in adulthood. However, adult sulcus PC arbors develop a greater number of branch crossings. These results suggest developmental neuronal plasticity that enables cerebellar PCs to attain correct functional adult architecture under different spatial constraints.

  1. First experimental evaluation of cardiac apex rotation with an epicardial coriolis force sensor.

    Science.gov (United States)

    Marcelli, Emanuela; Plicchi, Gianni; Cercenelli, Laura; Bortolami, Filippo

    2005-01-01

    Cardiac apex rotation, quantified by sophisticated techniques (radiopaque markers and tagged magnetic resonance), has been shown to provide a sensitive index of left ventricle (LV) dynamics. The authors describe the first experimental assessment of cardiac apex rotation using a gyroscopic sensor based on Coriolis force, epicardially glued on the apex. Dynamics of apex rotation were evaluated in a sheep at baseline, after a positive inotropic drug infusion, and after impairment of cardiac function induced by coronary ligation. To evaluate the efficacy of the sensor to monitor cardiac function, results were compared to contractility variations expressed by the maximum value of the first derivative of LV pressure (LVdP/dtMAX). After inotropic drug infusion, a parallel increasing trend resulted for LVdP/dtMAX, for the maximum value of angular velocity measured by the sensor, and for apex rotation angle derived from velocity signal (+146%, +155%, and +11% from baseline, respectively), whereas a decreasing trend of all three parameters resulted after coronary ligation (-35%, -31%, and -65%). The twist pattern also was altered from baseline. These initial results suggest that the use of an implantable rotation sensor based on Coriolis force can be an efficient and effective tool to assess LV torsional deformation both in normal and failing hearts.

  2. Accuracy of three electronic apex locators in the presence of different irrigating solutions

    Directory of Open Access Journals (Sweden)

    Ana Laura Pion Carvalho

    2010-12-01

    Full Text Available The present study compared the accuracy of three electronic apex locators (EALs - Elements Diagnostic®, Root ZX® and Apex DSP® - in the presence of different irrigating solutions (0.9% saline solution and 1% sodium hypochlorite. The electronic measurements were carried out by three examiners, using twenty extracted human permanent maxillary central incisors. A size 10 K file was introduced into the root canals until reaching the 0.0 mark, and was subsequently retracted to the 1.0 mark. The gold standard (GS measurement was obtained by combining visual and radiographic methods, and was set 1 mm short of the apical foramen. Electronic length values closer to the GS (± 0.5 mm were considered as accurate measures. Intraclass correlation coefficients (ICCs were used to verify inter-examiner agreement. The comparison among the EALs was performed using the McNemar and Kruskal-Wallis tests (p 0.05, independent of the irrigating solutions used. The measurements taken with these two EALs were more accurate than those taken with Apex DSP®, regardless of the irrigating solution used (p < 0.05. It was concluded that Elements Diagnostic® and Root ZX® apex locators are able to locate the cementum-dentine junction more precisely than Apex DSP®. The presence of irrigating solutions does not interfere with the performance of the EALs.

  3. Connecting tubule-selective knockout of AQP2 causes a mild urinary concentrating defect

    DEFF Research Database (Denmark)

    Kortenoeven, Marleen; Pedersen, Nis Borbye; Fenton, Robert A.

    . However, rat, mouse and humans were shown to express AQP2 in the CNT, which is regulated by vasopressin. Besides this, micropuncture studies showed a substantial water reabsorption in the CNT. To study the role of AQP2 in the CNT, AQP2-CNT-KO mice were generated by mating mice harboring loxP sites around...... exon 3 of the AQP2 gene with mice expressing Cre recombinase driven by the promoter region of the B1 subunit of V-ATPase. It was shown previously that this leads to Cre activity in 50% of the principal cells in the CNT. Knockout and wildtype mice were kept in metabolic cages for a total of 5 days...... groups (2616±188 mOsm/l in knockout animals vs. 2758±177 in the wildtype). Altogether, these data show that the AQP2-CNT-KO mice demonstrate a mild urinary concentrating defect. However, when challenged with an injection of dDAVP, the knockout mice were able to concentrate their urine to the same extent...

  4. Phenotypic Knockout of CXCR4 on Molt-4 with SDF-1α/54 Attached with KDEL

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Objective :To investigate the mechanism of phenotypic knockout of CXCR4 on T-cell leukemia cell line Molt-4 via SDF-1α/54/KDEL intrakine technology, which the mutant SDF-1α/54, human stromal cell-derived Faceor-1 (SDF-1α) was deleted its Cterminal α-helix and attached with a endoplasimc reticulum retention signal 4-peptide-KDEL encoding gene, so that retain the newly synthesized receptor CXCR4 within the Molt-4 cells endoplasmic reticulum. Methods: The recombinant vector pEGFP-C3/SDF-1α/54/KDEL were transfected into Cos-7 cells by liposome, SDF-1α/54/KDEL fusion protein was confirmed with western blot. The recombinant plasmids were transfected transiently into Molt-4 by electroporation. Results:Western blot confirmed SDF-1α/54/KDEL expression in Cos-7. A dramatic downregulation of CXCR4 expression on Molt-4 was demonstrated by flow cytometric (FCM) analysis. Conclusion:SDF-1α/54/KDEL and SDF-1αKDEL have no significant deviation for phenotypic knockout of CXCR4. These suggest that the phenotypic knockout effects of SDF-1α/54 against CXCR4 are not influenced by deleting of SDF-1α helix in the C-terminal.

  5. Comparative evaluation of accuracy of two electronic apex locators in the presence of various irrigants: An in vitro study

    OpenAIRE

    Saru Jain; Ravi Kapur

    2012-01-01

    Context: The establishment of appropriate working length is one of the most critical steps in endodontic therapy. Electronic apex locators have been introduced to determine the working length. The development of electronic apex locators has helped make the assessment of the working length more accurate and predictable, along with reduction in treatment time and radiation dose. Objectives: The aim of this study was to compare the efficacy of electronic apex locators after cleansing and shaping...

  6. Accuracy of electronic apex locator in length determination in the presence of different irrigants: An in vitro study

    OpenAIRE

    Soujanya; Muthu M; Sivakumar N

    2006-01-01

    Determining the root canal length accurately had been a challenge in endodontics. Introduction of apex locators have definitely served as an effective adjuvant to radiographs. This in vitro study had attempted to compare the effectiveness of electronic apex locator in the presence of various canal contents. Comparison of the length determined by apex locators with the radiographic length was also made. The results of the study had shown that the length of root canals can be accurately made ...

  7. Prohormone convertase 2 activity is increased in the hippocampus of Wfs1 knockout mice

    Directory of Open Access Journals (Sweden)

    Karin eTein

    2015-08-01

    Full Text Available BackgroundMutations in WFS1 gene cause Wolfram syndrome, which is a rare autosomal recessive disorder, characterized by diabetes insipidus, diabetes mellitus, optic nerve atrophy and deafness (DIDMOAD. The WFS1 gene product wolframin is located in the endoplasmic reticulum. Mice lacking this gene exhibit disturbances in the processing and secretion of peptides, such as vasopressin and insulin. In the brain, high levels of the wolframin protein have been observed in the hippocampus, amygdala and limbic structures. The aim of this study was to investigate the effect of Wfs1 knockout on peptide processing in mouse hippocampus. A peptidomic approach was used to characterize individual peptides in the hippocampus of wild-type and Wfs1 knockout mice. ResultsWe identified 126 peptides in hippocampal extracts and the levels of 10 peptides differed between Wfs1 KO and wild-type mice at P<0.05. The peptide with the largest alteration was little-LEN, which level was 25 times higher in the hippocampus of Wfs1 KO mice compared to wild-type mice. Processing (cleavage of little-LEN from the Pcsk1n gene product proSAAS involves prohormone convertase 2 (PC2. Thus, PC2 activity was measured in extracts prepared from the hippocampus of Wfs1 knockout mice. The activity of PC2 in Wfs1 mutant mice was significantly higher (149.9±2.3%, p<0.0001, n=8 than in wild-type mice (100.0±7.0%, n=8. However, Western blot analysis showed that protein levels of 7B2, proPC2 and PC2 were same in both groups, and so were gene expression levels.ConclusionsProcessing of proSAAS is altered in the hippocampus of Wfs1-KO mice, which is caused by increased activity of PC2. Increased activity of PC2 in Wfs1 knockout mice is not caused by alteration in the levels of PC2 protein. Our results suggest a functional link between Wfs1 and PC2. Thus, the detailed molecular mechanism of the role of Wfs1 in the regulation of PC2 activity needs further investigation.

  8. Impact of PI3Kγ gene knockout on acinar cells in mice with acute pancreatitis%磷脂酰肌醇3-激酶γ基因敲除对急性胰腺炎小鼠腺泡细胞的影响

    Institute of Scientific and Technical Information of China (English)

    贾文焯; 孙建华; 余涛; 肖刚

    2012-01-01

    目的:探讨磷脂酰肌醇3-激酶γ(PI3Kγ)基因敲除对急性胰腺炎(AP)小鼠病变程度、腺泡细胞功能以及对HSP70表达的影响.方法:雄性野生型( WT) C57BL/6小鼠和雄性PI3Kγ基因敲除(KO)小鼠各12只,随机均分为对照组和AP组,采用腹腔内注射蛙皮素诱导在体AP模型,对照组以同样的方式和体积给予生理盐水;另取两种动物各8只,行胰腺腺泡细胞体外分离,用胆囊收缩素(CCK-8)刺激作为离体AP模型,对照组给予二甲基亚砜(DMSO).观察胰腺组织病理变化、血清淀粉酶水平、胰腺组织和腺泡细胞胰蛋白酶活性和腺泡细胞淀粉酶释放率.Western blot检测胰腺组织及腺泡细胞中HSP70蛋白的表达.结果:病理学观察,两种小鼠的对照组胰腺未见异常,而AP组均出现不同程度的水肿、出血、坏死,经定量分析,KO小鼠胰腺炎腺泡细胞坏死数量和空泡数量明显少于WT小鼠(均P<0.05);两对照组间胰腺组织胰蛋白酶活性和体外腺泡细胞胰蛋白酶活性无明显差异(均P>0.05),但AP组(在体、离体)KO小鼠胰腺组织及腺泡的胰蛋白酶活性均低于WT小鼠(均P<0.05).两种小鼠间血清淀粉酶水平及腺泡细胞淀粉酶释放曲线差异无统计学意义(P>0.05);与对照组比较,AP组(在体、离体)小鼠胰腺组织和腺泡细胞HSP70表达均增加,且KO小鼠表达量明显大于WT小鼠(均P<0.05).结论:AP时,PI3Kγ可能通过下调HSP70表达水平和增强胰蛋白酶原活化促进腺泡细胞坏死,而对淀粉酶的分泌过程无明显影响.%Objective: To observe the effect of PI3Kγ gene knockout on the extent of the pancreatic lesion, acinar cellfunction and HSP70 protein expression level of mice with acute pancreatitis (AP).Methods: The male wild-type (WT) C57BL/6 mice and PI3Kγ gene knockout (KO) mice were randomly divided into control group and AP group with 12 mice in each group. The in vivo AP model was induced by intraperitoneal

  9. Effects of Akt1 gene knockout on pain behaviour induced by chronic constriction injury of sciatic nerve in mice%Akt1基因敲除对坐骨神经结扎小鼠痛行为的影响

    Institute of Scientific and Technical Information of China (English)

    隽立芹; 薄靳华; 马正良; 顾小萍

    2014-01-01

    目的 探讨Akt1基因敲除对坐骨神经结扎小鼠痛行为学的影响.方法 C57BL/6雄性小鼠随机分为Akt1基因敲除组(KO组,n=12)和野生组(WT组,n=12).在小鼠右侧制作坐骨神经慢性挤压(chronic constriction injury,CCI)模型,测试术前1d和术后1d、3d、5d、7d、10d、14d、17d、21 d的机械缩足阈值(paw withdrawal mechanical threshold,PWMT)和热缩足潜伏期(paw withdrawal thermal latency,PWTL).结果 KO组和WT组小鼠的两侧PWMT基础值[右侧:(0.89±0.15)g,(0.87±0.15)g;左侧:(0.97 ±0.19)g,(1.05±0,14)g,P>0.05]和PWTL[右侧:(7.64±0.71)s,(7.56±0.68)s;左侧:(7.67±0.6)s,(7.64±0.64)s,P>0.05]差异无统计学意义.术后各测试时间点KO组/WT组小鼠右侧的PWMT和PWTL与其基础值相比均明显减低(P<0.05),KO组左侧PWMT和PWTL与WT组小鼠相比差异无统计学意义(P>0.05),但是KO组右侧PWMT和PWTL较WT组明显降低(P<0.05).结论 Akt1基因敲除后会加重小鼠坐骨神经结扎所诱发的神经病理性疼痛.%Objective To investigate the effects of Aktl gene knockout on pain behavior induced by chronic constriction injury model of sciatic nerve (CCI).Methods C57BL/6 male mice were randomly divided into Akt1 knockout group (KO group,n=12),wild type group(WT group,n=12).All mice were made model of CCI in the right sciatic nerve.Each mouse received tests of the paw withdrawal mechanical threshold (PWMT) and the paw withdrawal thermal latency(PWTL) at the times of 1d before and 1 d,3 d,5 d,7 d,10 d,14 d,17 d,21 d after surgery.Results For both KO group and WT group,the basic values of PMWT(right(0.89±0.15)g,(0.87±0.15)g; left(0.97±0.19) g,(1.05±0.14) g,P>0.05) and PWTL(right (7.64±0.71) s,(7.56±0.68) s ;left: (7.67±0.6) s,(7.64±0.64) s,P>0.05) showed no significantly statistical difference.Compared with WT group and the basic value,PWMT and PWTL were significantly decreased after surgery in KO group (P<0.05).The PWMT and P WTL of the left paw in KO group

  10. Establishment and phenotypic analysis of an Mstn knockout rat.

    Science.gov (United States)

    Gu, Hao; Cao, Yong; Qiu, Bin; Zhou, Zhiqiang; Deng, Ran; Chen, Zhuang; Li, Rongfeng; Li, Xueling; Wei, Qiang; Xia, Xianzhu; Yong, Weidong

    2016-08-12

    Myostatin (Mstn) is an inhibitor of myogenesis, regulating the number and size of skeletal myocytes. In addition to its myogenic regulatory function, Mstn plays important roles in the development of adipose tissues and in metabolism. In the present study, an Mstn knockout rat model was generated using the zinc finger nuclease (ZFN) technique in order to further investigate the function and mechanism of Mstn in metabolism. The knockout possesses a frame shift mutation resulting in an early termination codon and a truncated peptide of 109 amino acids rather than the full 376 amino acids. The absence of detectable mRNA confirmed successful knockout of Mstn. Relative to wild-type (WT) littermates, Knockout (KO) rats exhibited significantly greater body weight, body circumference, and muscle mass. However, no significant differences in grip force was observed, indicating that Mstn deletion results in greater muscle mass but not greater muscle fiber strength. Additionally, KO rats were found to possess less body fat relative to WT littermates, which is consistent with previous studies in mice and cattle. The aforementioned results indicate that Mstn knockout increases muscle mass while decreasing fat content, leading to observed increases in body weight and body circumference. The Mstn knockout rat model provides a novel means to study the role of Mstn in metabolism and Mstn-related muscle hypertrophy. PMID:27289021

  11. Efficiency of 2 electronic apex locators on working length determination: A clinical study

    Directory of Open Access Journals (Sweden)

    Sibel Koçak

    2013-01-01

    Full Text Available Aims: The aim of this clinical study was to evaluate the clinical accuracy of two electronic apex locators (EALs. Materials and Methods: A total of 120 patients with 283 roots were randomized into three groups including, traditional radiographic method, EAL (Root ZX mini, and apex locating endodontic motor (VDW Gold for working length (WL determination. Root canals were instrumented to a size ProTaper F3 nickel titanium file. The obturation quality of matched tapered master cone (ProTaper F3 was determined for the accuracy of WL. Statistical Analysis Used: Descriptive statistics were expressed as numbers and percentages. Pearson Chi-square test was used to determine for differences between groups. P < 0.05 was considered statistically significant for all tests. Results: There was no statistically significant difference between the three tested groups ( P = 0.894. Conclusions: The success of both apex locators was similar to the radiographic WL determination technique.

  12. Aircraft emissions, plume chemistry, and alternative fuels: results from the APEX, AAFEX, and MDW-2009 campaigns

    Science.gov (United States)

    Wood, E. C.; Herndon, S. C.; Timko, M.; Yu, Z.; Miake-Lye, R. C.; Lee, B. H.; Santoni, G.; Munger, J. W.; Wofsy, S.; Anderson, B.; Knighton, W. B.

    2009-12-01

    We describe observations of aircraft emissions from the APEX, JETS-APEX2, APEX3, MDW-2009 and AAFEX campaigns. Direct emissions of HOx precursors are important for understanding exhaust plume chemistry due to their role in determining HOx concentrations. Nitrous acid (HONO) and formaldehyde are crucial HOx precursors and thus drivers of plume chemistry. At idle power, aircraft engine exhaust is unique among fossil fuel combustion sources due to the speciation of both NOx and VOCs. The impacts of emissions of HOx precursors on plume chemistry at low power are demonstrated with empirical observations of rapid NO to NO2 conversion, indicative of rapid HOx chemistry. The impacts of alternative fuels (derived from biomass, coal, and natural gas) on emissions of NOx, CO, and speciated VOCs are discussed.

  13. First 230 GHz VLBI Fringes on 3C 279 using the APEX Telescope

    OpenAIRE

    J. Wagner; Roy, A L; Krichbaum, T. P.; Alef, W.; Bansod, A.; Bertarini, A.; Güsten, R.; Graham, D.; HODGSON, J; Märtens, R.; Menten, K.; Muders, D.; Rottmann, H.; TUCCARI, G.; Weiss, A

    2015-01-01

    We report about a 230 GHz very long baseline interferometry (VLBI) fringe finder observation of blazar 3C 279 with the APEX telescope in Chile, the phased submillimeter array (SMA), and the SMT of the Arizona Radio Observatory (ARO). We installed VLBI equipment and measured the APEX station position to 1 cm accuracy (1 sigma). We then observed 3C 279 on 2012 May 7 in a 5 hour 230 GHz VLBI track with baseline lengths of 2800 M$\\lambda$ to 7200 M$\\lambda$ and a finest fringe spacing of 28.6 mic...

  14. A DESIGN FOR APEX FUSION REACTOR MODEL BY USING MONTE CARLO METHOD

    OpenAIRE

    Şarer, Başar; HANÇERLİOĞLU, Aybaba; SAVRUK, Nurettin

    2010-01-01

    ABSTRACTIn APEX fusion reactor model, the first solid wall has been replaced with fast flowing thin liquid wall layer. Liquid wall has a large potential to enhance the vision of fusion. Because, liquid wall usage can increase life time of the structure to that of the reactor by decreasing failures on the structural materials, and also allows high neutron wall load (>10 MW/m2) (1), (2), (3), (10). In this study, design and calculations of APEX were carried out as 3-D torus by using MCNP- 4B...

  15. FLASH(+) —A Dual-Channel Wide-Band Spectrometer for APEX

    Science.gov (United States)

    Klein, Thomas; Ciechanowicz, Miroslaw; Leinz, Christian; Heyminck, Stefan; Gusten, Rolf; Kasemann, Christoph; Wunsch, Joachim; Maier, Doris; Sekimoto, Yutaro

    2014-09-01

    We report on the design and on-site performance of the recently upgraded FLASH+ instrument operated at the APEX telescope in the Chilean Atacama desert in 5100 m altitude. The dual-channel receiver allows parallel observations in the atmospheric submillimeter windows between 268 and 516 GHz. A signal of in total 16 GHz is processed simultaneously. Equipped with state-of-the-art sideband separating mixers (spin-off developments from the ALMA bands 7 and 8), its outstanding on-sky performance makes FLASH+ a most efficient spectral line mapping machine. Operation is fully automated and allows flexible remote observations from the APEX base in San Pedro de Atacama.

  16. Behavioral analysis of male and female Fmr1 knockout mice on C57BL/6 background

    OpenAIRE

    Ding, Qi; Sethna, Ferzin; Wang, Hongbing

    2014-01-01

    Fragile X syndrome (FXS) is a monogenic disease caused by mutations in the FMR1 gene. The Fmr1 knockout (KO) mice show many aspects of FXS-related phenotypes, and have been used as a major pre-clinical model for FXS. Although FXS occurs in both male and female patients, most studies on the mouse model use male animals. Few studies test whether gender affects the face validity of the mouse model. Here, we examined multiple behavioral phenotypes with male hemizygous and female homozygous Fmr1 K...

  17. Impaired conditioned taste aversion learning in spinophilin knockout mice.

    Science.gov (United States)

    Stafstrom-Davis, C A; Ouimet, C C; Feng, J; Allen, P B; Greengard, P; Houpt, T A

    2001-01-01

    Plasticity in dendritic spines may underlie learning and memory. Spinophilin, a protein enriched in dendritic spines, has the properties of a scaffolding protein and is believed to regulate actin cytoskeletal dynamics affecting dendritic spine morphology. It also binds protein phosphatase-1 (PP-1), an enzyme that regulates dendritic spine physiology. In this study, we tested the role of spinophilin in conditioned taste aversion learning (CTA) using transgenic spinophilin knockout mice. CTA is a form of associative learning in which an animal rejects a food that has been paired previously with a toxic effect (e.g., a sucrose solution paired with a malaise-inducing injection of lithium chloride). Acquisition and extinction of CTA was tested in spinophilin knockout and wild-type mice using taste solutions (sucrose or sodium chloride) or flavors (Kool-Aid) paired with moderate or high doses of LiCl (0.15 M, 20 or 40 mL/kg). When sucrose or NaCl solutions were paired with a moderate dose of LiCl, spinophilin knockout mice were unable to learn a CTA. At the higher dose, knockout mice acquired a CTA but extinguished more rapidly than wild-type mice. A more salient flavor stimulus (taste plus odor) revealed similar CTA learning at both doses of LiCl in both knockouts and wild types. Sensory processing in the knockouts appeared normal because knockout mice and wild-type mice expressed identical unconditioned taste preferences in two-bottle tests, and identical lying-on-belly responses to acute LiCl. We conclude that spinophilin is a candidate molecule required for normal CTA learning. PMID:11584074

  18. Targeting cancer using KAT inhibitors to mimic lethal knockouts

    Science.gov (United States)

    Brown, James A.L.; Bourke, Emer; Eriksson, Leif A.; Kerin, Michael J.

    2016-01-01

    Two opposing enzyme classes regulate fundamental elements of genome maintenance, gene regulation and metabolism, either through addition of an acetyl moiety by histone acetyltransferases (HATs) or its removal by histone de-acetyltransferases (HDAC), and are exciting targets for drug development. Importantly, dysfunctional acetylation has been implicated in numerous diseases, including cancer. Within the HAT superfamily the MYST family holds particular interest, as its members are directly involved in the DNA damage response and repair pathways and crucially, several members have been shown to be down-regulated in common cancers (such as breast and prostate). In the present study we focus on the development of lysine (K) acetyltransferase inhibitors (KATi) targeting the MYST family member Tip60 (Kat5), an essential protein, designed or discovered through screening libraries. Importantly, Tip60 has been demonstrated to be significantly down-regulated in many cancers which urgently require new treatment options. We highlight current and future efforts employing these KATi as cancer treatments and their ability to synergize and enhance current cancer treatments. We investigate the different methods of KATi production or discovery, their mechanisms and their validation models. Importantly, the utility of KATi is based on a key concept: using KATi to abrogate the activity of an already down-regulated essential protein (effectively creating a lethal knockout) provides another innovative mechanism for targeting cancer cells, while significantly minimizing any off-target effects to normal cells. This approach, combined with the rapidly developing interest in KATi, suggests that KATi have a bright future for providing truly personalized therapies. PMID:27528742

  19. Knockout of the abundant Trichomonas vaginalis hydrogenosomal membrane protein TvHMP23 increases hydrogenosome size but induces no compensatory up-regulation of paralogous copies.

    Science.gov (United States)

    Brás, Xavier Pereira; Zimorski, Verena; Bolte, Kathrin; Maier, Uwe-G; Martin, William F; Gould, Sven B

    2013-05-01

    The Trichomonas vaginalis genome encodes up to 60000 genes, many of which stem from genome duplication events. Paralogous copies thus accompany most T. vaginalis genes, a phenomenon that limits genetic manipulation. We characterized one of the parasite's most abundant hydrogenosomal membrane proteins, TvHMP23, which is phylogenetically distinct from canonical metabolite carriers, and which localizes to the inner hydrogenosomal membrane as shown through sub-organellar fractionation and protease protection assays. Knockout of Tvhmp23 through insertion of the selectable neomycin marker led to a size increase of hydrogenosomes, the first knockout-induced phenotypes reported for Trichomonas, but no growth impairment. The transcriptional response of its four paralogous copies then analyzed revealed that they are not up-regulated, and hence do not compensate for the Tvhmp23 knockout. PMID:23499435

  20. From Nf1 to Sdhb knockout: Successes and failures in the quest for animal models of pheochromocytoma.

    Science.gov (United States)

    Lepoutre-Lussey, Charlotte; Thibault, Constance; Buffet, Alexandre; Morin, Aurélie; Badoual, Cécile; Bénit, Paule; Rustin, Pierre; Ottolenghi, Chris; Janin, Maxime; Castro-Vega, Luis-Jaime; Trapman, Jan; Gimenez-Roqueplo, Anne-Paule; Favier, Judith

    2016-02-01

    Pheochromocytomas and paragangliomas (PPGL) are rare neuroendocrine tumors characterized by a high frequency of hereditary forms. Based on transcriptome classification, PPGL can be classified in two different clusters. Cluster 1 tumors are caused by mutations in SDHx, VHL and FH genes and are characterized by a pseudohypoxic signature. Cluster 2 PPGL carry mutations in RET, NF1, MAX or TMEM127 genes and display an activation of the MAPK and mTOR signaling pathways. Many genetically engineered and allografted mouse models have been generated these past 30 years to investigate the mechanisms of PPGL tumorigenesis and test new therapeutic strategies. Among them, only Cluster 2-related models have been successful while no Cluster 1-related knockout mouse was so far reported to develop a PPGL. In this review, we present an overview of existing, successful or not, PPGL models, and a description of our own experience on the quest of Sdhb knockout mouse models of PPGL.

  1. FMR1基因敲除小鼠脑组织微白蛋白表达的改变及其意义%Changes and significance of expression of parvalbumin in brain tissues of FMR1 gene knockout mice

    Institute of Scientific and Technical Information of China (English)

    邸伟; 易咏红; 曾志涌; 徐明明; 王玉良; 孙卫文; 廖卫平

    2010-01-01

    目的 探讨微白蛋白(PV)阳性中间神经元在脆性X综合征(FXS)癫痫易感性增加中的作用. 方法 应用免疫组织化学染色检测FVB近交系雄性2、4、6 W龄FMR1基因敲除型(KO)(KO2W、KO4W、KO6W)和同龄野生型(WT)(WT2W、WT4W、WT6W)小鼠大脑纹状皮质、颞听皮质、梨状皮质及海马CA1区、CA3区、齿状回中PV的表达(n=6);应用Western blot法检测上述小鼠大脑皮层、海马组织PV的含量(n=6). 结果 KO2W、KO44W小鼠的大脑纹状皮质、颞听皮质、梨状皮质、海马CA1和CA3区PV阳性中间神经元的数量分别较WT2W、WT4-小鼠减少,差异有统计学意义(P<0.05);KO2W和KO4W小鼠大脑皮层、海马中PV含量分别较WT2W、WT4W小鼠减少,差异有统计学意义(P<0.05). 结论 PV阳性中间神经元及PV含量的减少.可能是引起FXS模型鼠癫痫易感性增加的主要原因.%Objective To explore the possible role of parvalbumin (PV)-positive interneuron in the pathogenesis of increased susceptibility to epileptic seizures in FMR1 gene knockout (FMR1 KO)mice. Methods Immunohistochemistry was employed to determine the expression of PV in CA1 and CA3 regions of the hippocampus, the striate cortex, the temporal auditory cortex and the piriform cortex of FVB strain FMR1 KO mice and wild type (WT) controls at the age of 2, 4 and 6 w. Western blotting was used to detect the level of PV in the cerebral cortex and hippocampus of the above mice. Results The numbers of PV-positive interneuron in CA1 and CA3 regions of the hippocampus, the striate cortex,the temporal auditory cortex and the piriform cortex of FMR1 KO mice at the age of 2 and 4 w were significantly decreased as compared with those in the age-matched WT mice (P<0.05). The level of PV in the cerebral cortex and hippocampus in FMR1 KO mice at the age of 2 and 4 w was also significantly decreased than that in the age-matched WT mice (P<0.05). Conclusion Decreased numbers of PV-positive interneuron and level of PV

  2. 载脂蛋白E基因敲除及高脂饮食小鼠皮质额叶区形态学及Mortalin表达的变化%Change of apolipoprotein E gene knock-out and high-fat diet on morphology and mortalin expression in mice frontal lobe neurons

    Institute of Scientific and Technical Information of China (English)

    杨平; 周祎; 刘娟; 黄大可; 桂丽; 汪渊; 贾雪梅

    2011-01-01

    目的 观察载脂蛋白E基因敲除(ApoE KO)及高脂饮食(HF)后小鼠额叶区形态学及Mortalin蛋白表达变化,以进一步探讨ApoE异常导致阿尔茨海默病及记忆损伤的可能机制.方法 10只野生型小鼠予普通饲料喂养作为对照(C)组,10只ApoE KO小鼠予普通饲料喂养作为KO组,10只ApoE KO小鼠予高脂饲料喂养作为KO-HF组.小鼠造膜成功后称重,取血检测血脂,取小鼠脑组织石蜡包埋切片,分别进行HE染色、尼氏染色、免疫组化染色及计算机图像分析.结果 KO组体质量、总胆固醇、甘油三酯及低密度脂蛋白胆固醇含量与C组相比明显升高,KO-HF组升高更明显(P<0.05);KO组及KO-HF组小鼠大脑皮质额叶神经元内尼氏体平均光密度值较C组减少,Mortalin平均光密度值较C组升高(P<0.01).结论 ApoE KO及HF可致额叶区神经元内尼氏体减少,Mortalin蛋白表达上调,该蛋白可能与ApoE异常导致阿尔茨海默症认知功能障碍有着密切的关系.%Objective To investigate the effect of apolipoprotein E gene knock-out( ApoE KO )and high-fat diet ( HF ) on morphology and the expression of mortalin in neurons of mice cortex frontal lobe, and explore the impact of these factors on memorv and Alzheimer ’ s disease . Methods Ten wild- type and 10 ApoE KO mice were fed with common chow as the control ( C ) group and the KO group respectively , while 10 ApoE KO mice were fed with HF as the KO-HF group. 12 weeks later.the weight and the lid of these mice were measured. The brain tissues were observed by HE staining, nissl staining,immunohistochemistry staining and image analysis by computer. Results In the ApoE KO group , weight , total cholesterol, triglyceride .low-density lipoprotein cholesterol were higher than those in the C group, and these changes were more significant in KO-HF group. The average optical density of the nissl body was higher in the KO group and in KO-HF group than those in the C group( P <0. 01

  3. Expression of neuregulin 1 and its significance in brain tissues of FMR1 gene knockout mice%Fmr1基因敲除小鼠脑组织神经调节蛋白1表达的改变及其意义

    Institute of Scientific and Technical Information of China (English)

    卢韬; 欧阳梅; 周林涛; 易咏红

    2013-01-01

    目的 明确神经调节蛋白1 (NRG1)在Fmr1基因敲除(KO)小鼠中的变化,探讨其在脆性X综合征发病机制中的作用. 方法 应用免疫组织化学染色法检测FVB近交系雄性2周龄Fmr1 KO小鼠(KO2w)、4周龄Fmr1 KO小鼠(KO4w)和同龄野生型(WT)小鼠大脑皮层及海马CA1区、CA3区、齿状回中神经调节蛋白1的阳性神经元的数量;Western blotting检测上述小鼠大脑皮层和海马组织NRG1蛋白的含量. 结果 与同龄WT小鼠相比,KO2w、KO4w小鼠大脑皮层、海马CA1和CA3区NRG1阳性神经元的数量明显减少,在海马齿状回却明显增多,差异均有统计学意义(P<0.05);KO2w、KO4w小鼠大脑皮层、海马中NRG1含量(相对分子质量为55 000亚型)分别较同龄的WT小鼠明显减少,差异亦有统计学意义(P<0.05). 结论 Fmr1 KO小鼠大脑皮层和海马组织NRG1阳性神经元及NRG1蛋白表达明显减少,NRG1可能参与脆性X综合征发病机制.%Objective To explore the expression changes ofneuregulin 1 (NRG1) in Fmr1 gene knockout (Fmr1 KO) mice,and its possible role in fragile X syndrome pathogenesis.Methods Immunohistochemistry and Western blotting were simultaneously employed to detect the expression levels of NRG 1 in the hippocampus and the cortex of FVB strain Fmr1 KO mice and wild type (WT) controls at the age of 2 and 4 weeks.Results The number of NRG1-positive cells in the CA1 and CA3 regions of hippocampus and the cortex of Fmr1 KO mice at the age of 2 and 4 weeks was significantly smaller than that in the age-matched WT mice (P<0.05).The NRG1 protein levels in the hippocampus and cerebral cortex of Fmr1 KO mice at the age of 2 and 4 weeks were also decreased as compared with those in the age-matched WT mice (P<0.05).Conclusion The number of NRG1-positive cells and NRG1 protein levels in the CA1 and CA3 regions of hippocampus and the cortex of Fmr1 KO mice are decreased,indicating that NRG 1 might involve in the pathogenesis of Fragile X

  4. 糖基转移酶Colgalt2基因敲除对肝细胞再生过程中增殖和凋亡的作用研究%Effect of glycosyltransferase Colgalt2 gene knockout on hepatocyte proliferation and apoptosis on course of hepatocyte regeneration

    Institute of Scientific and Technical Information of China (English)

    王智强; 杨琪; 王建文; 刘燃; 郝晓花; 黄玉波; 魏红山

    2015-01-01

    Objective To investigate the effect of Colgalt2 gene knockout on hepatocyte proliferation and apoptosis on course of hepatocyte regeneration.Methods Colgalt2+/+wild type control mice and Colgalt2-/-mice models of 70% partial hepatectomy (PH) were established according to the protocols by Higgins and Anderson in 1931. Then the part of right lateral lobes of liver from normal mice and those 1, 3, 7 days after PH were harvested, ifxed in 10% neutral buffered formalin, embedded in parafifn and cut into tissue sections. Immunohistochemistry was used to compare the expressive difference of proliferating cell nuclear antigen (PCNA) in the mouse liver slices at different time points after partial hepatectomy. Primary hepatocytes of normal Colgalt2+/+wild type control mice, normal Colgalt2-/- mice and those treated by partial hepatectomyafter 1, 3, 5 days were isolated and puriifed by two steps of collagenase perfusion. Flow cytometry was used to detect apoptotic rate in each group.Results The number of PCNA positive cells of Colgalt2-/-mice was much larger than that of Colgalt2+/+mice on 1, 3, 7 days after PH, and the difference was statistically signiifcant (P<0.01). The viability of primary hepatocytes was 94%. The apoptotic rate of normal Colgalt2+/+ mice was (23.36 ±0.83)%, and that of normal Colgalt2-/-mice was (21.04 ±1.16)%. The apoptotic rates of Colgalt2+/+control mice and Colgalt2-/-mice in the ifrst day after PH were (8.27 ±0.33)%, (8.44 ±0.30)%. The apoptotic rates declined to the lowest on the ifrst day after PH and then they began to increase gradually. While, the apoptotic rates of Colgalt2+/+ mice and Colgalt2-/- mice in the third day after PH were (15.92 ±0.56)%, (12.14 ± 0.37)%. The apoptotic rate of Colgalt2-/- mice was much lower than that of wild type control mice (P<0.01). The apoptotic rates of Colgalt2+/+ mice and Colgalt2-/- mice in the iffth day after PH were (21.36 ±0.51)%, (18.92 ±0.92)% which showed that the apoptotic rate of Colgalt2

  5. Human Bacterial Artificial Chromosome (BAC) Transgenesis Fully Rescues Noradrenergic Function in Dopamine β-Hydroxylase Knockout Mice

    OpenAIRE

    Cubells, Joseph F.; Schroeder, Jason P; Barrie, Elizabeth S.; Manvich, Daniel F.; Wolfgang Sadee; Tiina Berg; Kristina Mercer; Stowe, Taylor A.; L Cameron Liles; Katherine E Squires; Andrew Mezher; Patrick Curtin; Dannie L Perdomo; Patricia Szot; David Weinshenker

    2016-01-01

    Dopamine β-hydroxylase (DBH) converts dopamine (DA) to norepinephrine (NE) in noradrenergic/adrenergic cells. DBH deficiency prevents NE production and causes sympathetic failure, hypotension and ptosis in humans and mice; DBH knockout (Dbh -/-) mice reveal other NE deficiency phenotypes including embryonic lethality, delayed growth, and behavioral defects. Furthermore, a single nucleotide polymorphism (SNP) in the human DBH gene promoter (-970C>T; rs1611115) is associated with variation in s...

  6. The transcobalamin receptor knockout mouse: a model for vitamin B12 deficiency in the central nervous system

    OpenAIRE

    Lai, Shao-Chiang; Nakayama, Yasumi; Sequeira, Jeffrey M.; Wlodarczyk, Bogdan J.; Cabrera, Robert M.; Finnell, Richard H.; Bottiglieri, Teodoro; Quadros, Edward V.

    2013-01-01

    The membrane receptor (TCblR/CD320) for transcobalamin (TC)-bound cobalamin (Cbl) facilitates the cellular uptake of Cbl. A genetically modified mouse model involving ablation of the CD320 gene was generated to study the effects on cobalamin homeostasis. The nonlethal nature of this knockout and the lack of systemic cobalamin deficiency point to other mechanisms for cellular Cbl uptake in the mouse. However, severe cobalamin depletion in the central nervous system (CNS) after birth (P

  7. Generation of ER{alpha}-floxed and knockout mice using the Cre/LoxP system

    Energy Technology Data Exchange (ETDEWEB)

    Antonson, P., E-mail: per.antonson@ki.se [Department of Biosciences and Nutrition, Karolinska Institutet, Novum, SE-141 83 Huddinge (Sweden); Omoto, Y.; Humire, P. [Department of Biosciences and Nutrition, Karolinska Institutet, Novum, SE-141 83 Huddinge (Sweden); Gustafsson, J.-A. [Department of Biosciences and Nutrition, Karolinska Institutet, Novum, SE-141 83 Huddinge (Sweden); Center for Nuclear Receptors and Cell Signaling, Department of Biology and Biochemistry, University of Houston, Houston, TX 77204 (United States)

    2012-08-10

    Highlights: Black-Right-Pointing-Pointer ER{alpha} floxed and knockout mice were generated. Black-Right-Pointing-Pointer Disruption of the ER{alpha} gene results in sterility in both male and female mice. Black-Right-Pointing-Pointer ER{alpha}{sup -/-} mice have ovaries with hemorrhagic follicles and hypoplastic uterus. Black-Right-Pointing-Pointer Female ER{alpha}{sup -/-} mice develop obesity. -- Abstract: Estrogen receptor alpha (ER{alpha}) is a nuclear receptor that regulates a range of physiological processes in response to estrogens. In order to study its biological role, we generated a floxed ER{alpha} mouse line that can be used to knock out ER{alpha} in selected tissues by using the Cre/LoxP system. In this study, we established a new ER{alpha} knockout mouse line by crossing the floxed ER{alpha} mice with Cre deleter mice. Here we show that genetic disruption of the ER{alpha} gene in all tissues results in sterility in both male and female mice. Histological examination of uterus and ovaries revealed a dramatically atrophic uterus and hemorrhagic cysts in the ovary. These results suggest that infertility in female mice is the result of functional defects of the reproductive tract. Moreover, female knockout mice are hyperglycemic, develop obesity and at the age of 4 months the body weight of these mice was more than 20% higher compared to wild type littermates and this difference increased over time. Our results demonstrate that ER{alpha} is necessary for reproductive tract development and has important functions as a regulator of metabolism in females.

  8. Petrous apex chordoma - a case report; Cordoma de apice petroso - relato de um caso

    Energy Technology Data Exchange (ETDEWEB)

    Silveira, Claudio Regis S. [Clinica Boghos Boyadjian, Fortaleza, CE (Brazil); Barreto, Cristina Marques; Rossi, Luis Antonio [Hospital do Servidor Publico Estadual de Sao Paulo, SP (Brazil). Servico de Radiologia; Michiloski, Custodio; Rotta, Jose Marcus [Hospital do Servidor Publico Estadual de Sao Paulo, SP (Brazil). Servico de Neurocirurgia; Almeida, Serguey Malaquias de

    2001-02-01

    Chordomas are rare neoplasms arising from notochordal remnants that persist along the axial skeleton. Intracranial chordomas occur more frequently in the midline. We describe a typical case of an off-midline chordoma arising from the petrous apex, and discuss the embryogenic factors which determine that location, as well as the symptoms, imaging findings, surgical treatment and evolution. (author)

  9. Code modernization and modularization of APEX and SWAT watershed simulation models

    Science.gov (United States)

    SWAT (Soil and Water Assessment Tool) and APEX (Agricultural Policy / Environmental eXtender) are respectively large and small watershed simulation models derived from EPIC Environmental Policy Integrated Climate), a field-scale agroecology simulation model. All three models are coded in FORTRAN an...

  10. 76 FR 13663 - Cooper Tools, Currently Known as Apex Tool Group, LLC, Hicksville, OH; Amended Certification...

    Science.gov (United States)

    2011-03-14

    ..., applicable to workers of Cooper Tools, Hicksville, Ohio. The workers are engaged in activities related to the production. The notice was published in the Federal Register on May 28, 2010 (75 FR 30069). At the request of... Employment and Training Administration Cooper Tools, Currently Known as Apex Tool Group, LLC, Hicksville,...

  11. Congenital cholesteatoma of petrous apex: Rare case report: Diagnostic and management challenge

    Directory of Open Access Journals (Sweden)

    Arun Dehadaray

    2013-01-01

    Full Text Available A rare case of congenital cholesteatoma of petrous apex with facial nerve palsy and its successful management is reported. 49 year old female presented with progressive vertigo since 2 years. Patient developed tinnitus and hearing loss in the right ear since 7 months and right sided complete facial asymmetry since 6 months. She had normal right tympanic membrane and complete right lower motor neuron facial nerve palsy. She also had profound sensorineural hearing loss with positive Cerebellar signs. Magnetic resonance imaging and High resolution computed tomography with contrast temporal bone showed extensive bony destruction and petrous apex lesion. Facial nerve and vestibular cochlear nerve was compressed by abnormal soft-tissue present in the internal auditory meatus. Transmastoid translabyrinthine exploration was carried out for petrous apex lesion. Intra-operative extensive bony erosion was noted in the temporal bone. Erosion was extending upto Internal Acoustic Meatus compressing VII and VIII nerve bundle. Post-operatively patient was relieved of vertigo and tinnitus. Though hearing didn′t improve, but there was an improvement in facial palsy. Congenital petrous apex cholesteatoma is very rare case. With no specific radiological signs congenital cholesteatoma is difficult to diagnose pre-operatively. It was a challenge to treat surgically such a rare case with extensive neurosurgical presentation without any neurological deficit. Patient showed improvement official nerve after the 1΍ year of surgery.

  12. APEX (Air Pollution Exercise) Volume 21: Legal References: Air Pollution Control Regulations.

    Science.gov (United States)

    Environmental Protection Agency, Research Triangle Park, NC. Office of Manpower Development.

    The Legal References: Air Pollution Control Regulations Manual is the last in a set of 21 manuals (AA 001 009-001 029) used in APEX (Air Pollution Exercise), a computerized college and professional level "real world" game simulation of a community with urban and rural problems, industrial activities, and air pollution difficulties. The manual…

  13. APEX (Air Pollution Exercise) Volume 8: Industrialist's Manual No. 3, Rusty's Iron Foundry.

    Science.gov (United States)

    Environmental Protection Agency, Research Triangle Park, NC. Office of Manpower Development.

    The Industrialist's Manual No. 3, Rusty's Iron Foundry is part of a set of 21 manuals (AA 001 009-001 029) used in APEX (Air Pollution Exercise), a computerized college and professional level "real world" game simulation of a community with urban and rural problems, industrial activities, and air pollution difficulties. The first two sections,…

  14. Time-domain multiplexed SQUID readout of a bolometer camera for APEX

    Energy Technology Data Exchange (ETDEWEB)

    Zakosarenko, V; Schulz, M; Krueger, A; Heinz, E; Anders, S; Peiselt, K; May, T; Meyer, H-G [Institute of Photonic Technology, Albert-Einstein-Strasse 9, 07745 Jena (Germany); Kreysa, E; Siringo, G; Esch, W [Max-Planck-Institute for Radioastronomy, Auf dem Huegel 69, 53121 Bonn (Germany); Starkloff, M, E-mail: zakosarenko@ipht-jena.de [Supracon AG, Wildenbruchstrasse 15, 07745 Jena (Germany)

    2011-01-15

    We describe the development and fabrication of the sub-millimeter APEX (Atacama pathfinder experiment) bolometer camera (SABOCA) and its installation at the APEX radio telescope in Chile. SABOCA has 37 bolometers for a wavelength of about 0.35 mm. The bolometers with superconducting transition-edge sensors are read out by superconducting quantum interference devices (SQUIDs). Time-domain multiplexing was used to compress 40 readout signals into four output channels, which are further amplified by superconducting quantum interference filters (SQIF). The multiplexer readout circuit is described in detail. One spare SQUID and two spare bolometers have been used as reference to evaluate contributions to the bolometer noise figure. SABOCA's electrical noise equivalent power is about 7 x 10{sup -16} W Hz{sup -1/2}, whereas the optical noise equivalent flux density measured at APEX is better than 0.35 Jy Hz{sup -1/2} (1 Jy = 10{sup -26} W m{sup -2} Hz{sup -1}). The measured bolometer noise exceeds the theoretical estimation by factor of two, possibly caused by excess noise in the transition-edge sensor. Nevertheless the achieved instrumental sensitivity meets the specification given by the APEX board. SABOCA was commissioned as the facility instrument in the spring of 2009.

  15. Minimally invasive surgical method to detect sound processing in the cochlear apex by optical coherence tomography

    Science.gov (United States)

    Ramamoorthy, Sripriya; Zhang, Yuan; Petrie, Tracy; Fridberger, Anders; Ren, Tianying; Wang, Ruikang; Jacques, Steven L.; Nuttall, Alfred L.

    2016-02-01

    Sound processing in the inner ear involves separation of the constituent frequencies along the length of the cochlea. Frequencies relevant to human speech (100 to 500 Hz) are processed in the apex region. Among mammals, the guinea pig cochlear apex processes similar frequencies and is thus relevant for the study of speech processing in the cochlea. However, the requirement for extensive surgery has challenged the optical accessibility of this area to investigate cochlear processing of signals without significant intrusion. A simple method is developed to provide optical access to the guinea pig cochlear apex in two directions with minimal surgery. Furthermore, all prior vibration measurements in the guinea pig apex involved opening an observation hole in the otic capsule, which has been questioned on the basis of the resulting changes to cochlear hydrodynamics. Here, this limitation is overcome by measuring the vibrations through the unopened otic capsule using phase-sensitive Fourier domain optical coherence tomography. The optically and surgically advanced method described here lays the foundation to perform minimally invasive investigation of speech-related signal processing in the cochlea.

  16. Apex simulation: environmental benefits of agroforestry and grass buffers for corn-soybean watersheds

    Science.gov (United States)

    The Agricultural Policy Environmental Extender (APEX) model is used to simulate the effects of vegetative filter strips on runoff and pollutant loadings from agricultural watersheds. A long-term paired watershed study under corn (Zea mays L-soybean [Glycine max (L.) Merr.] rotation with agroforestr...

  17. In vitro selection of rape variants resistant to oxalic acid using haploid stem apexes

    International Nuclear Information System (INIS)

    Mutagenic treatment was made of the haploid stem apexes rape strain '9841' and '9885' with Pingyangmycin. As a result of positive selection with oxalic acid providing selection pressure, variants with significantly higher tolerance to oxalic acid than the original ones were obtained. 3 germplasm with significantly higher resistance to Sclerotinia sclerotiorum than cultivar Hu You 12 were selected from field test

  18. Impact of Apex Model parameterization strategy on estimated benefit of conservation practices

    Science.gov (United States)

    Three parameterized Agriculture Policy Environmental eXtender (APEX) models for corn-soybean rotation on clay pan soils were developed with the objectives, 1. Evaluate model performance of three parameterization strategies on a validation watershed; and 2. Compare predictions of water quality benefi...

  19. Time-domain multiplexed SQUID readout of a bolometer camera for APEX

    International Nuclear Information System (INIS)

    We describe the development and fabrication of the sub-millimeter APEX (Atacama pathfinder experiment) bolometer camera (SABOCA) and its installation at the APEX radio telescope in Chile. SABOCA has 37 bolometers for a wavelength of about 0.35 mm. The bolometers with superconducting transition-edge sensors are read out by superconducting quantum interference devices (SQUIDs). Time-domain multiplexing was used to compress 40 readout signals into four output channels, which are further amplified by superconducting quantum interference filters (SQIF). The multiplexer readout circuit is described in detail. One spare SQUID and two spare bolometers have been used as reference to evaluate contributions to the bolometer noise figure. SABOCA's electrical noise equivalent power is about 7 x 10-16 W Hz-1/2, whereas the optical noise equivalent flux density measured at APEX is better than 0.35 Jy Hz-1/2 (1 Jy = 10-26 W m-2 Hz-1). The measured bolometer noise exceeds the theoretical estimation by factor of two, possibly caused by excess noise in the transition-edge sensor. Nevertheless the achieved instrumental sensitivity meets the specification given by the APEX board. SABOCA was commissioned as the facility instrument in the spring of 2009.

  20. Metabolomic Characterization of Knockout Mutants in Arabidopsis: Development of a Metabolite Profiling Database for Knockout Mutants in Arabidopsis1[W][OPEN

    Science.gov (United States)

    Fukushima, Atsushi; Kusano, Miyako; Mejia, Ramon Francisco; Iwasa, Mami; Kobayashi, Makoto; Hayashi, Naomi; Watanabe-Takahashi, Akiko; Narisawa, Tomoko; Tohge, Takayuki; Hur, Manhoi; Wurtele, Eve Syrkin; Nikolau, Basil J.; Saito, Kazuki

    2014-01-01

    Despite recent intensive research efforts in functional genomics, the functions of only a limited number of Arabidopsis (Arabidopsis thaliana) genes have been determined experimentally, and improving gene annotation remains a major challenge in plant science. As metabolite profiling can characterize the metabolomic phenotype of a genetic perturbation in the plant metabolism, it provides clues to the function(s) of genes of interest. We chose 50 Arabidopsis mutants, including a set of characterized and uncharacterized mutants, that resemble wild-type plants. We performed metabolite profiling of the plants using gas chromatography-mass spectrometry. To make the data set available as an efficient public functional genomics tool for hypothesis generation, we developed the Metabolite Profiling Database for Knock-Out Mutants in Arabidopsis (MeKO). It allows the evaluation of whether a mutation affects metabolism during normal plant growth and contains images of mutants, data on differences in metabolite accumulation, and interactive analysis tools. Nonprocessed data, including chromatograms, mass spectra, and experimental metadata, follow the guidelines set by the Metabolomics Standards Initiative and are freely downloadable. Proof-of-concept analysis suggests that MeKO is highly useful for the generation of hypotheses for genes of interest and for improving gene annotation. MeKO is publicly available at http://prime.psc.riken.jp/meko/. PMID:24828308

  1. Reassessing the trophic role of reef sharks as apex predators on coral reefs

    Science.gov (United States)

    Frisch, Ashley J.; Ireland, Matthew; Rizzari, Justin R.; Lönnstedt, Oona M.; Magnenat, Katalin A.; Mirbach, Christopher E.; Hobbs, Jean-Paul A.

    2016-06-01

    Apex predators often have strong top-down effects on ecosystem components and are therefore a priority for conservation and management. Due to their large size and conspicuous predatory behaviour, reef sharks are typically assumed to be apex predators, but their functional role is yet to be confirmed. In this study, we used stomach contents and stable isotopes to estimate diet, trophic position and carbon sources for three common species of reef shark ( Triaenodon obesus, Carcharhinus melanopterus and C. amblyrhynchos) from the Great Barrier Reef (Australia) and evaluated their assumed functional role as apex predators by qualitative and quantitative comparisons with other sharks and large predatory fishes. We found that reef sharks do not occupy the apex of coral reef food chains, but instead have functional roles similar to those of large predatory fishes such as snappers, emperors and groupers, which are typically regarded as high-level mesopredators. We hypothesise that a degree of functional redundancy exists within this guild of predators, potentially explaining why shark-induced trophic cascades are rare or subtle in coral reef ecosystems. We also found that reef sharks participate in multiple food webs (pelagic and benthic) and are sustained by multiple sources of primary production. We conclude that large conspicuous predators, be they elasmobranchs or any other taxon, should not axiomatically be regarded as apex predators without thorough analysis of their diet. In the case of reef sharks, our dietary analyses suggest they should be reassigned to an alternative trophic group such as high-level mesopredators. This change will facilitate improved understanding of how reef communities function and how removal of predators (e.g., via fishing) might affect ecosystem properties.

  2. One-neutron knockout from Ne24-28 isotopes

    CERN Document Server

    Rodriguez-Tajes, C; Caamano, M; Faestermann, T; Cortina-Gil, D; Zhukov, M; Simon, H; Nilsson, T; Borge, M J G; Alvarez-Pol, H; Winkler, M; Prochazka, A; Nociforo, C; Weick, H; Kanungo, R; Perez-Loureiro, D; Kurtukian, T; Suemmerer, K; Eppinger, K; Perea, A; Chatillon, A; Maierbeck, P; Benlliure, J; Pascual-Izarra, C; Gernhaeuser, R; Geissel, H; Aumann, T; Kruecken, R; Larsson, K; Tengblad, O; Benjamim, E; Jonson, B; Casarejos, E

    2010-01-01

    One-neutron knockout reactions of Ne24-28 in a beryllium target have been studied in the Fragment Separator (FRS), at GSI. The results include inclusive one-neutron knockout cross-sections as well as longitudinal-momentum distributions of the knockout fragments. The ground-state structure of the neutron-rich neon isotopes was obtained from an analysis of the measured momentum distributions. The results indicate that the two heaviest isotopes, Ne-27 and Ne-28, are dominated by a configuration in which a s(1/2) neutron is coupled to an excited state of the Ne-26 and Ne-27 core, respectively. (C) 2010 Elsevier B.V. All rights reserved.

  3. Functional deficits in PAK5, PAK6 and PAK5/PAK6 knockout mice.

    Directory of Open Access Journals (Sweden)

    Melody A Furnari

    Full Text Available The p21-activated kinases are effector proteins for Rho-family GTPases. PAK4, PAK5, and PAK6 are the group II PAKs associated with neurite outgrowth, filopodia formation, and cell survival. Pak4 knockout mice are embryonic lethal, while Pak5, Pak6, and Pak5/Pak6 double knockout mice are viable and fertile. Our previous work found that the double knockout mice exhibit locomotor changes and learning and memory deficits. We also found some differences with Pak5 and Pak6 single knockout mice and the present work further explores the potential differences of the Pak5 knockout and Pak6 knockout mice in comparison with wild type mice. The Pak6 knockout mice were found to weigh significantly more than the other genotypes. The double knockout mice were found to be less active than the other genotypes. The Pak5 knockout mice and the double knockout mice performed worse on the rotorod test. All the knockout genotypes were found to be less aggressive in the resident intruder paradigm. The double knockout mice were, once again, found to perform worse in the active avoidance assay. These results indicate, that although some behavioral differences are seen in the Pak5 and Pak6 single knockout mice, the double knockout mice exhibit the greatest changes in locomotion and learning and memory.

  4. CRISPR-Mediated VHL Knockout Generates an Improved Model for Metastatic Renal Cell Carcinoma.

    Science.gov (United States)

    Schokrpur, Shiruyeh; Hu, Junhui; Moughon, Diana L; Liu, Peijun; Lin, Lucia C; Hermann, Kip; Mangul, Serghei; Guan, Wei; Pellegrini, Matteo; Xu, Hua; Wu, Lily

    2016-01-01

    Metastatic renal cell carcinoma (mRCC) is nearly incurable and accounts for most of the mortality associated with RCC. Von Hippel Lindau (VHL) is a tumour suppressor that is lost in the majority of clear cell RCC (ccRCC) cases. Its role in regulating hypoxia-inducible factors-1α (HIF-1α) and -2α (HIF-2α) is well-studied. Recent work has demonstrated that VHL knock down induces an epithelial-mesenchymal transition (EMT) phenotype. In this study we showed that a CRISPR/Cas9-mediated knock out of VHL in the RENCA model leads to morphologic and molecular changes indicative of EMT, which in turn drives increased metastasis to the lungs. RENCA cells deficient in HIF-1α failed to undergo EMT changes upon VHL knockout. RNA-seq revealed several HIF-1α-regulated genes that are upregulated in our VHL knockout cells and whose overexpression signifies an aggressive form of ccRCC in the cancer genome atlas (TCGA) database. Independent validation in a new clinical dataset confirms the upregulation of these genes in ccRCC samples compared to adjacent normal tissue. Our findings indicate that loss of VHL could be driving tumour cell dissemination through stabilization of HIF-1α in RCC. A better understanding of the mechanisms involved in this phenomenon can guide the search for more effective treatments to combat mRCC. PMID:27358011

  5. Sirh7/Ldoc1 knockout mice exhibit placental P4 overproduction and delayed parturition.

    Science.gov (United States)

    Naruse, Mie; Ono, Ryuichi; Irie, Masahito; Nakamura, Kenji; Furuse, Tamio; Hino, Toshiaki; Oda, Kanako; Kashimura, Misho; Yamada, Ikuko; Wakana, Shigeharu; Yokoyama, Minesuke; Ishino, Fumitoshi; Kaneko-Ishino, Tomoko

    2014-12-01

    Sirh7/Ldoc1 [sushi-ichi retrotransposon homolog 7/leucine zipper, downregulated in cancer 1, also called mammalian retrotransposon-derived 7 (Mart7)] is one of the newly acquired genes from LTR retrotransposons in eutherian mammals. Interestingly, Sirh7/Ldoc1 knockout (KO) mice exhibited abnormal placental cell differentiation/maturation, leading to an overproduction of placental progesterone (P4) and placental lactogen 1 (PL1) from trophoblast giant cells (TGCs). The placenta is an organ that is essential for mammalian viviparity and plays a major endocrinological role during pregnancy in addition to providing nutrients and oxygen to the fetus. P4 is an essential hormone in the preparation and maintenance of pregnancy and the determination of the timing of parturition in mammals; however, the biological significance of placental P4 in rodents is not properly recognized. Here, we demonstrate that mouse placentas do produce P4 in mid-gestation, coincident with a temporal reduction in ovarian P4, suggesting that it plays a role in the protection of the conceptuses specifically in this period. Pregnant Sirh7/Ldoc1 knockout females also displayed delayed parturition associated with a low pup weaning rate. All these results suggest that Sirh7/Ldoc1 has undergone positive selection during eutherian evolution as a eutherian-specific acquired gene because it impacts reproductive fitness via the regulation of placental endocrine function. PMID:25468940

  6. Antibodies directed against monomorphic and evolutionary conserved self epitopes may be generated in 'knock-out' mice. Development of monoclonal antibodies directed against monomorphic MHC class I determinants

    DEFF Research Database (Denmark)

    Claesson, M H; Endel, B; Ulrik, J;

    1994-01-01

    Beta-2 microglobulin (beta 2m) gene 'knock-out' mice (C1D) were primed with purified H-2Kb and H-2Db molecules and spleen cells from immunized mice were used to generate monoclonal antibody secreting B-cell hybridomas. Approximately 0.2% of the Ig-secreting primary microcultures contained H-2b...

  7. Multigene knockout utilizing off-target mutations of the CRISPR/Cas9 system in rice.

    Science.gov (United States)

    Endo, Masaki; Mikami, Masafumi; Toki, Seiichi

    2015-01-01

    The clustered regularly interspaced short palindromic repeat (CRISPR)-associated endonuclease 9 (CRISPR/Cas9) system has been demonstrated to be a robust genome engineering tool in a variety of organisms including plants. However, it has been shown that the CRISPR/Cas9 system cleaves genomic DNA sequences containing mismatches to the guide RNA strand. We expected that this low specificity could be exploited to induce multihomeologous and multiparalogous gene knockouts. In the case of polyploid plants, simultaneous modification of multiple homeologous genes, i.e. genes with similar but not identical DNA sequences, is often needed to obtain a desired phenotype. Even in diploid plants, disruption of multiparalogous genes, which have functional redundancy, is often needed. To validate the applicability of the CRISPR/Cas9 system to target mutagenesis of paralogous genes in rice, we designed a single-guide RNA (sgRNA) that recognized 20 bp sequences of cyclin-dependent kinase B2 (CDKB2) as an on-target locus. These 20 bp possess similarity to other rice CDK genes (CDKA1, CDKA2 and CDKB1) with different numbers of mismatches. We analyzed mutations in these four CDK genes in plants regenerated from Cas9/sgRNA-transformed calli and revealed that single, double and triple mutants of CDKA2, CDKB1 and CDKB2 can be created by a single sgRNA.

  8. CyclinD(-/-)/Patched(+/-)双基因敲除鼠的构建%Construction of CyclinD(-/-)/Patched(+/-)double knockout mice

    Institute of Scientific and Technical Information of China (English)

    辛世杰; 李璇; Molly Duman-Scheel; Wei Du

    2008-01-01

    目的 构建CycD(-/-)/Ptc(+/-)双基因敲除鼠以验证Hh传导系统与Rb系统的关系.方法 应用已有的杂合子CycD基因敲除鼠CycD(+/-)与杂合子Ptc基因敲除鼠Ptc(+/-)进行交配繁殖,得到双基因杂合子敲除鼠CycD(+/-)/Ptc(+/-),再利用该双基因杂合子敲除鼠进行二次交配繁殖,得到实验组CycD(-/-)/Ptc(+/-)双基因敲除鼠.结果 实验得到了双基因敲除鼠CycD(-/-)/Ptc(+/-)、双基因杂合子基因敲除鼠CycD(+/-)/ptc(+/-)、cycD单基因敲除鼠CycD(-/-)/Ptc(+/+).结论 通过本实验方法得到CycD(-/-)/Ptc(+/-)双基因敲除鼠,可用于进一步揭示Ptc基因与CycD基因相互影响调控的作用及Hedgehog与Rb信号传导通路在哺乳动物中的相关性.%Objective To construct CyclinD(CycD)(-/-)/Patched(Ptc)(+/-)double knockout mice to stuay the relationship between Hedgehog and Rb signaling pathways.Methods Amphimixis was performed between female heterozygote CycD knockout mice CycD(+/-)and male heterozygote Ptc knockout mice Ptc(+/-),thus CycD(+/-)/Ptc(+/-)double knockout mice were obtained.These mice underwent secondary matched and thus construct CycD(-/-)/Ptc(+/-)double knockout mice.Results CycD(-/-)/Ptc(+/-)double knockout mice,CycD(+/-)/Ptc(+/-) heterozygote double knockout mice, and CycD knockout mice CycD( -/-)/Ptc( +/+) were gained.Conclusion The method of amphimixis can construct CycD (-/-)/Ptc (+/-)double knockout mice that can be used to disclose the regulation between Ptc gene and CycD gene and the relativity between Hedgehog and Rb pathways in mammals.

  9. Knockout mutants as a tool to identify the subunit composition of Arabidopsis glutamine synthetase isoforms.

    Science.gov (United States)

    Dragićević, Milan; Todorović, Slađana; Bogdanović, Milica; Filipović, Biljana; Mišić, Danijela; Simonović, Ana

    2014-06-01

    Glutamine synthetase (GS) is a key enzyme in nitrogen assimilation, which catalyzes the formation of glutamine from ammonia and glutamate. Plant GS isoforms are multimeric enzymes, recently shown to be decamers. The Arabidopsis genome encodes five cytosolic (GS1) proteins labeled as GLN1;1 through GLN1;5 and one chloroplastic (GS2) isoform, GLN2;0. However, as many as 11 GS activity bands were resolved from different Arabidopsis tissues by Native PAGE and activity staining. Western analysis showed that all 11 isoforms are composed exclusively of 40 kDa GS1 subunits. Of five GS1 genes, only GLN1;1, GLN1;2 and GLN1;3 transcripts accumulated to significant levels in vegetative tissues, indicating that only subunits encoded by these three genes produce the 11-band zymogram. Even though the GS2 gene also had significant expression, the corresponding activity was not detected, probably due to inactivation. To resolve the subunit composition of 11 active GS1 isoforms, homozygous knockout mutants deficient in the expression of different GS1 genes were selected from the progeny of T-DNA insertional SALK and SAIL lines. Comparison of GS isoenzyme patterns of the selected GS1 knockout mutants indicated that all of the detected isoforms consist of varying proportions of GLN1;1, GLN1;2 and GLN1;3 subunits, and that GLN1;1 and GLN1;3, as well as GLN1;2 and GLN1;3 and possibly GLN1;1 and GLN1;2 proteins combine in all proportions to form active homo- and heterodecamers.

  10. Threshold Energies for Single Carbon Knockout from Polycyclic Aromatic Hydrocarbons

    CERN Document Server

    Stockett, M H; Chen, T; de Ruette, N; Giacomozzi, L; Wolf, M; Schmidt, H T; Zettergren, H; Cederquist, H

    2015-01-01

    We have measured absolute cross sections for ultrafast (fs) single-carbon knockout from Polycyclic Aromatic Hydrocarbon (PAH) cations as functions of He-PAH center-of-mass collision energy in the range 10-200 eV. Classical Molecular Dynamics (MD) simulations cover this range and extend up to 10$^5$ eV. The shapes of the knockout cross sections are well described by a simple analytical expression yielding experimental and MD threshold energies of $E_{th}^{Exp}=32.5\\pm 0.4$ eV and $E_{th}^{MD}=41.0\\pm 0.3$ eV, respectively. These are the first measurements of knockout threshold energies for molecules isolated \\emph{in vacuo}. We further deduce semi-empirical (SE) and MD displacement energies --- \\emph{i.e.} the energy transfers to the PAH molecules at the threshold energies for knockout --- of $T_{disp}^{SE}=23.3\\pm 0.3$ eV and $T_{disp}^{MD}=27.0\\pm 0.3$ eV. The semi-empirical results compare favorably with measured displacement energies for graphene $T_{disp}=23.6$ eV [Meyer \\emph{et al.} Phys. Rev Lett. \\tex...

  11. Cardiomyocyte-specific conditional knockout of the histone chaperone HIRA in mice results in hypertrophy, sarcolemmal damage and focal replacement fibrosis.

    Science.gov (United States)

    Valenzuela, Nicolas; Fan, Qiying; Fa'ak, Faisal; Soibam, Benjamin; Nagandla, Harika; Liu, Yu; Schwartz, Robert J; McConnell, Bradley K; Stewart, M David

    2016-03-01

    HIRA is the histone chaperone responsible for replication-independent incorporation of histone variant H3.3 within gene bodies and regulatory regions of actively transcribed genes, and within the bivalent promoter regions of developmentally regulated genes. The HIRA gene lies within the 22q11.2 deletion syndrome critical region; individuals with this syndrome have multiple congenital heart defects. Because terminally differentiated cardiomyocytes have exited the cell cycle, histone variants should be utilized for the bulk of chromatin remodeling. Thus, HIRA is likely to play an important role in epigenetically defining the cardiac gene expression program. In this study, we determined the consequence of HIRA deficiency in cardiomyocytes in vivo by studying the phenotype of cardiomyocyte-specific Hira conditional-knockout mice. Loss of HIRA did not perturb heart development, but instead resulted in cardiomyocyte hypertrophy and susceptibility to sarcolemmal damage. Cardiomyocyte degeneration gave way to focal replacement fibrosis and impaired cardiac function. Gene expression was widely altered in Hira conditional-knockout hearts. Significantly affected pathways included responses to cellular stress, DNA repair and transcription. Consistent with heart failure, fetal cardiac genes were re-expressed in the Hira conditional knockout. Our results suggest that transcriptional regulation by HIRA is crucial for cardiomyocyte homeostasis. PMID:26935106

  12. The transcobalamin receptor knockout mouse: a model for vitamin B12 deficiency in the central nervous system.

    Science.gov (United States)

    Lai, Shao-Chiang; Nakayama, Yasumi; Sequeira, Jeffrey M; Wlodarczyk, Bogdan J; Cabrera, Robert M; Finnell, Richard H; Bottiglieri, Teodoro; Quadros, Edward V

    2013-06-01

    The membrane receptor (TCblR/CD320) for transcobalamin (TC)-bound cobalamin (Cbl) facilitates the cellular uptake of Cbl. A genetically modified mouse model involving ablation of the CD320 gene was generated to study the effects on cobalamin homeostasis. The nonlethal nature of this knockout and the lack of systemic cobalamin deficiency point to other mechanisms for cellular Cbl uptake in the mouse. However, severe cobalamin depletion in the central nervous system (CNS) after birth (Pmouse model; however, it does provide a model with which to evaluate metabolic pathways and genes affected. PMID:23430977

  13. Hmga1/Hmga2 double knock-out mice display a “superpygmy” phenotype

    Directory of Open Access Journals (Sweden)

    Antonella Federico

    2014-04-01

    Full Text Available The HMGA1 and HMGA2 genes code for proteins belonging to the High Mobility Group A family. Several genes are negatively or positively regulated by both these proteins, but a number of genes are specifically regulated by only one of them. Indeed, knock-out of the Hmga1 and Hmga2 genes leads to different phenotypes: cardiac hypertrophy and type 2 diabetes in the former case, and a large reduction in body size and amount of fat tissue in the latter case. Therefore, to better elucidate the functions of the Hmga genes, we crossed Hmga1-null mice with mice null for Hmga2. The Hmga1−/−/Hmga2−/− mice showed reduced vitality and a very small size (75% smaller than the wild-type mice; they were even smaller than pygmy Hmga2-null mice. The drastic reduction in E2F1 activity, and consequently in the expression of the E2F-dependent genes involved in cell cycle regulation, likely accounts for some phenotypic features of the Hmga1−/−/Hmga2−/− mice.

  14. Altered neurocircuitry in the dopamine transporter knockout mouse brain.

    Directory of Open Access Journals (Sweden)

    Xiaowei Zhang

    Full Text Available The plasma membrane transporters for the monoamine neurotransmitters dopamine, serotonin, and norepinephrine modulate the dynamics of these monoamine neurotransmitters. Thus, activity of these transporters has significant consequences for monoamine activity throughout the brain and for a number of neurological and psychiatric disorders. Gene knockout (KO mice that reduce or eliminate expression of each of these monoamine transporters have provided a wealth of new information about the function of these proteins at molecular, physiological and behavioral levels. In the present work we use the unique properties of magnetic resonance imaging (MRI to probe the effects of altered dopaminergic dynamics on meso-scale neuronal circuitry and overall brain morphology, since changes at these levels of organization might help to account for some of the extensive pharmacological and behavioral differences observed in dopamine transporter (DAT KO mice. Despite the smaller size of these animals, voxel-wise statistical comparison of high resolution structural MR images indicated little morphological change as a consequence of DAT KO. Likewise, proton magnetic resonance spectra recorded in the striatum indicated no significant changes in detectable metabolite concentrations between DAT KO and wild-type (WT mice. In contrast, alterations in the circuitry from the prefrontal cortex to the mesocortical limbic system, an important brain component intimately tied to function of mesolimbic/mesocortical dopamine reward pathways, were revealed by manganese-enhanced MRI (MEMRI. Analysis of co-registered MEMRI images taken over the 26 hours after introduction of Mn(2+ into the prefrontal cortex indicated that DAT KO mice have a truncated Mn(2+ distribution within this circuitry with little accumulation beyond the thalamus or contralateral to the injection site. By contrast, WT littermates exhibit Mn(2+ transport into more posterior midbrain nuclei and contralateral

  15. Generation of a Knockout Mouse Embryonic Stem Cell Line Using a Paired CRISPR/Cas9 Genome Engineering Tool.

    Science.gov (United States)

    Wettstein, Rahel; Bodak, Maxime; Ciaudo, Constance

    2016-01-01

    CRISPR/Cas9, originally discovered as a bacterial immune system, has recently been engineered into the latest tool to successfully introduce site-specific mutations in a variety of different organisms. Composed only of the Cas9 protein as well as one engineered guide RNA for its functionality, this system is much less complex in its setup and easier to handle than other guided nucleases such as Zinc-finger nucleases or TALENs.Here, we describe the simultaneous transfection of two paired CRISPR sgRNAs-Cas9 plasmids, in mouse embryonic stem cells (mESCs), resulting in the knockout of the selected target gene. Together with a four primer-evaluation system, it poses an efficient way to generate new independent knockout mouse embryonic stem cell lines.

  16. Effects of Dissolving Solutions on the Accuracy of an Electronic Apex Locator-Integrated Endodontic Handpiece

    Directory of Open Access Journals (Sweden)

    Yakup Ustun

    2013-01-01

    Full Text Available The effects of three dissolving agents on the accuracy of an electronic apex locator- (EAL- integrated endodontic handpiece during retreatment procedures were evaluated. The true lengths (TLs of 56 extracted incisor teeth were determined visually. Twenty teeth were filled with gutta-percha and a resin-based sealer (group A, 20 with gutta-percha and a zinc oxide/eugenol-based sealer (group B, and 16 roots were used as the control group (group C. All roots were prepared to TL. Guttasolv, Resosolv, and Endosolv E were used as the dissolving solutions. Two evaluations of the handpiece were performed: the apical accuracy during the auto reverse function (ARL and the apex locator function (EL alone. The ARL function of the handpiece gave acceptable results. There were significant differences between the EL mode measurements and the TL (P<0.05. In these comparisons, Tri Auto ZX EL mode measurements were significantly shorter than those of the TL.

  17. Results from the NRC AP600 testing program at the Oregon State University APEX facility

    Energy Technology Data Exchange (ETDEWEB)

    Reyes, J.N. Jr. [Oregon State Univ., Corvallis, OR (United States); Bessette, D.E. [Nuclear regulatory Systems, Washington, DC (United States); DiMarzo, M. [Univ. of Maryland, College Park, MD (United States)

    1996-03-01

    The Department of Nuclear Engineering at Oregon State University (OSU) is performing a series of confirmatory tests for the U.S. Nuclear Regulatory Commission. These tests are being conducted in the Advanced Plant Experiment (APEX) facility which is a 1/4 length scale and 1/192 volume scale integral system simulation of the Westinghouse Advanced Passive 600 MWe (AP600) plant. The purpose of the testing program is to examine AP600 passive safety system performance, particularly during long term cooling. Thus far, OSU has successfully performed ten integral system tests for the NRC. This paper presents a description of the APEX facility and summarizes the important results of the NRC test program at OSU.

  18. Persistent scarring and dilated cardiomyopathy suggest incomplete regeneration of the apex resected neonatal mouse myocardium

    DEFF Research Database (Denmark)

    Andersen, Ditte Caroline; Jensen, Charlotte Harken; Baun, Christina;

    2016-01-01

    exhibited decreased ejection fraction and wall motion with increased end-diastolic and systolic volumes compared to sham-operated mice. Scarring with collagen accumulation was still substantial, with increased heart size, while cardiomyocyte size was unaffected. In conclusion, these data thus show that apex......Heart damage in mammals is generally considered to result in scar formation, whereas zebrafish completely regenerate their hearts following an intermediate and reversible state of fibrosis after apex resection (AR). Recently, using the AR procedure, one-day-old mice were suggested to have full...... capacity for cardiac regeneration as well. In contrast, using the same mouse model others have shown that the regeneration process is incomplete and that scarring still remains 21days after AR. The present study tested the hypothesis that like in zebrafish, fibrosis in neonatal mammals could...

  19. Accuracy of a fourth generation apex locator-an in vitro evaluation

    Directory of Open Access Journals (Sweden)

    Dalia Abdullah

    2007-09-01

    Full Text Available The new fourth generation electronic apex locator (EAL, Elements (SybronEndo, USA has been introduced recently in the market. This study aims to investigate the accuracy of this EAL and to compare the result with a well-known apex locator, Root ZX and the radiographic technique using an in vitro model. Thirty anterior teeth with straight canals stored in 10% formalin were used. Access cavities were prepared followed by coronal flaring of the canals. Water was used as an irrigant. After the actual lengths (AL were measured, the teeth were then embedded in an alginate model. Periapical radiograph of each tooth was taken using a digital sensor and the radiographic lengths (RL were measured 0.5 mm short of the radiographic apex. Electronic tooth length measurements (EL were carried out using both EAL. Canals were then irrigated with 2.5% NaOCl and EL was taken again. Results showed that both EALs were highly accurate to within + 0.5 mm of the apical foramen, with mean differences between the AL and EL of Elements 0.23mm (SE = 0.04 and Root ZX was 0.31mm (SE = 0.05. RL was significantly less accurate compared to the readings from both EAL. No significant difference was found in the reading between both apex locators when measurements were taken in NaOCl solution. Both Elements and Propex proved to be as reliable as Root ZX. Presence of sodium hypochlorite solution did not affect the accuracy of the measurements.

  20. First 230 GHz VLBI fringes on 3C 279 using the APEX Telescope

    Science.gov (United States)

    Wagner, J.; Roy, A. L.; Krichbaum, T. P.; Alef, W.; Bansod, A.; Bertarini, A.; Güsten, R.; Graham, D.; Hodgson, J.; Märtens, R.; Menten, K.; Muders, D.; Rottmann, H.; Tuccari, G.; Weiss, A.; Wieching, G.; Wunderlich, M.; Zensus, J. A.; Araneda, J. P.; Arriagada, O.; Cantzler, M.; Duran, C.; Montenegro-Montes, F. M.; Olivares, R.; Caro, P.; Bergman, P.; Conway, J.; Haas, R.; Johansson, J.; Lindqvist, M.; Olofsson, H.; Pantaleev, M.; Buttaccio, S.; Cappallo, R.; Crew, G.; Doeleman, S.; Fish, V.; Lu, R.-S.; Ruszczyk, C.; SooHoo, J.; Titus, M.; Freund, R.; Marrone, D.; Strittmatter, P.; Ziurys, L.; Blundell, R.; Primiani, R.; Weintroub, J.; Young, K.; Bremer, M.; Sánchez, S.; Marscher, A. P.; Chilson, R.; Asada, K.; Inoue, M.

    2015-09-01

    Aims: We report about a 230 GHz very long baseline interferometry (VLBI) fringe finder observation of blazar 3C 279 with the APEX telescope in Chile, the phased submillimeter array (SMA), and the SMT of the Arizona Radio Observatory (ARO). Methods: We installed VLBI equipment and measured the APEX station position to 1 cm accuracy (1σ). We then observed 3C 279 on 2012 May 7 in a 5 h 230 GHz VLBI track with baseline lengths of 2800 Mλ to 7200 Mλ and a finest fringe spacing of 28.6 μas. Results: Fringes were detected on all baselines with signal-to-noise ratios of 12 to 55 in 420 s. The correlated flux density on the longest baseline was ~0.3 Jy beam-1, out of a total flux density of 19.8 Jy. Visibility data suggest an emission region ≲ 38 μas in size, and at least two components, possibly polarized. We find a lower limit of the brightness temperature of the inner jet region of about 1010 K. Lastly, we find an upper limit of 20% on the linear polarization fraction at a fringe spacing of ~ 38 μas. Conclusions: With APEX the angular resolution of 230 GHz VLBI improves to 28.6 μas. This allows one to resolve the last-photon ring around the Galactic Center black hole event horizon, expected to be 40 μas in diameter, and probe radio jet launching at unprecedented resolution, down to a few gravitational radii in galaxies like M 87. To probe the structure in the inner parsecs of 3C 279 in detail, follow-up observations with APEX and five other mm-VLBI stations have been conducted (March 2013) and are being analyzed.

  1. Determining the Biogenicity of Microfossils in the Apex Chert, Western Australia, Using Transmission Electron Microscopy

    Science.gov (United States)

    DeGregorio, B. T.; Sharp, T. G.

    2003-01-01

    For over a decade, the oldest evidence for life on this planet has been microfossils in the 3.5 Ga Apex Chert in Western Australia. Recently, the biogenicity of these carbon-rich structures has been called into question through reanalysis of the local geology and reinterpretation of the original thin sections. Although initially described as a stratiform, bedded chert of siliceous clasts, the unit is now thought to be a brecciated hydrothermal vein chert. The high temperatures of a hydrothermal environment would probably have detrimental effects to early non-hyperthermophilic life, compared to that of a shallow sea. Conversely, a hydrothermal origin would suggest that if the microfossils were valid, they might have been hyperthermophilic. Apex Chert controversy. The Apex Chert microfossils were originally described as septate filaments composed of kerogen similar in morphology to Proterozoic and modern cyanobacteria. However new thin section analysis shows that these carbonaceous structures are not simple filaments. Many of the original microfossils are branched and have variable thickness when the plane of focus is changed. Hydrothermal alteration of organic remains has also been suggested for the creation of these strange morphologies. Another point of contention lies with the nature of the carbon material in these proposed microfossils. Kerogen is structurally amorphous, but transforms into well-ordered graphite under high pressures and temperatures. Raman spectrometry of the carbonaceous material in the proposed microfossils has been interpreted both as partially graphitized kerogen and amorphous graphite. However, these results are inconclusive, since Raman spectrometry cannot adequately discriminate between kerogen and disordered graphite. There are also opposing views for the origin of the carbon in the Apex Chert. The carbon would be biogenic if the proposed microfossils are indeed the remains of former living organisms. However, an inorganic Fischer

  2. High resolution NO2 remote sensing from the Airborne Prism EXperiment (APEX imaging spectrometer

    Directory of Open Access Journals (Sweden)

    B. Buchmann

    2012-03-01

    Full Text Available We present and evaluate the retrieval of high spatial resolution maps of NO2 vertical column densities (VCD from the Airborne Prism EXperiment (APEX imaging spectrometer. APEX is a novel instrument providing airborne measurements of unique spectral and spatial resolution and coverage as well as high signal stability. In this study, we use spectrometer data acquired over Zurich, Switzerland, in the morning and late afternoon during a flight campaign on a cloud-free summer day in June 2010. NO2 VCD are derived with a two-step approach usually applied to satellite NO2 retrievals, i.e. a DOAS analysis followed by air mass factor calculations based on radiative transfer computations. Our analysis demonstrates that APEX is clearly sensitive to NO2 VCD above typical European tropospheric background abundances (>1 × 1015 molec cm−2. The two-dimensional maps of NO2 VCD reveal a very plausible spatial distribution with strong gradients around major NOx sources (e.g. Zurich airport, waste incinerator, motorways and low NO2 in remote areas. The morning overflights resulted in generally higher NO2 VCD and a more distinct pattern than the afternoon overflights which can be attributed to the meteorological conditions prevailing during that day (development of the boundary layer and increased wind speed in the afternoon as well as to photochemical loss of NO2. The remotely sensed NO2 VCD are also highly correlated with ground-based in-situ measurements from local and national air quality networks (R=0.73. Airborne NO2 remote sensing using APEX will be valuable to detect NO2 emission sources, to provide input for NO2 emission modeling, and to establish links between in-situ measurements, air quality models, and satellite NO2 products.

  3. High-resolution NO2 remote sensing from the Airborne Prism EXperiment (APEX imaging spectrometer

    Directory of Open Access Journals (Sweden)

    B. Buchmann

    2012-09-01

    Full Text Available We present and evaluate the retrieval of high spatial resolution maps of NO2 vertical column densities (VCD from the Airborne Prism EXperiment (APEX imaging spectrometer. APEX is a novel instrument providing airborne measurements of unique spectral and spatial resolution and coverage as well as high signal stability. In this study, we use spectrometer data acquired over Zurich, Switzerland, in the morning and late afternoon during a flight campaign on a cloud-free summer day in June 2010. NO2 VCD are derived with a two-step approach usually applied to satellite NO2 retrievals, i.e. a DOAS analysis followed by air mass factor calculations based on radiative transfer computations. Our analysis demonstrates that APEX is clearly sensitive to NO2 VCD above typical European tropospheric background abundances (>1 × 1015 molec cm−2. The two-dimensional maps of NO2 VCD reveal a very convincing spatial distribution with strong gradients around major NOx sources (e.g. Zurich airport, waste incinerator, motorways and low NO2 in remote areas. The morning overflights resulted in generally higher NO2 VCD and a more distinct pattern than the afternoon overflights which can be attributed to the meteorological conditions prevailing during that day with stronger winds and hence larger dilution in the afternoon. The remotely sensed NO2 VCD are also in reasonably good agreement with ground-based in-situ measurements from air quality networks considering the limitations of comparing column integrals with point measurements. Airborne NO2 remote sensing using APEX will be valuable to detect NO2 emission sources, to provide input for NO2 emission modelling, and to establish links between in-situ measurements, air quality models, and satellite NO2 products.

  4. Usefulness of Image Guidance in the Surgical Treatment of Petrous Apex Cholesterol Granuloma

    Directory of Open Access Journals (Sweden)

    A. Pietrantonio

    2013-01-01

    Full Text Available The petrous apex is a pyramid-shaped structure, located medial to the inner ear and the intrapetrous segment of the internal carotid artery. Lesions of the petrous apex can be surgically treated through different surgical routes. Because of the important neurovascular structures located inside the temporal bone, anatomical 3D knowledge is paramount. For this reason, image-guided surgery could represent a useful tool. We report the case of a young woman who came to our observation for a trigeminal neuralgia due to a petrous apex cholesterol granuloma. The lesion was treated through the placement of a drainage tube via an infracochlear approach, with the aid of neuronavigation and intraoperative MRI. Preoperative CT scan images and intraoperative MRI images were fused for surgical planning. The accuracy of the neuronavigation system has proved to be good, and the safety of the procedure was enhanced. Therefore, neuronavigation and intraoperative MRI, though not available in all neurootological centres, should be considered useful tools in these challenging procedures.

  5. First 230 GHz VLBI Fringes on 3C 279 using the APEX Telescope

    CERN Document Server

    Wagner, J; Krichbaum, T P; Alef, W; Bansod, A; Bertarini, A; Güsten, R; Graham, D; Hodgson, J; Märtens, R; Menten, K; Muders, D; Rottmann, H; Tuccari, G; Weiss, A; Wieching, G; Wunderlich, M; Zensus, J A; Araneda, J P; Arriagada, O; Cantzler, M; Duran, C; Montenegro-Montes, F M; Olivares, R; Caro, P; Bergman, P; Conway, J; Haas, R; Johansson, J; Lindqvist, M; Olofsson, H; Pantaleev, M; Buttaccio, S; Cappallo, R; Crew, G; Doeleman, S; Fish, V; Lu, R -S; Ruszczyk, C; SooHoo, J; Titus, M; Freund, R; Marrone, D; Strittmatter, P; Ziurys, L; Blundell, R; Primiani, R; Weintroub, J; Young, K; Bremer, M; Sánchez, S; Marscher, A P; Chilson, R; Asada, K; Inoue, M

    2015-01-01

    We report about a 230 GHz very long baseline interferometry (VLBI) fringe finder observation of blazar 3C 279 with the APEX telescope in Chile, the phased submillimeter array (SMA), and the SMT of the Arizona Radio Observatory (ARO). We installed VLBI equipment and measured the APEX station position to 1 cm accuracy (1 sigma). We then observed 3C 279 on 2012 May 7 in a 5 hour 230 GHz VLBI track with baseline lengths of 2800 M$\\lambda$ to 7200 M$\\lambda$ and a finest fringe spacing of 28.6 micro-arcseconds. Fringes were detected on all baselines with SNRs of 12 to 55 in 420 s. The correlated flux density on the longest baseline was ~0.3 Jy/beam, out of a total flux density of 19.8 Jy. Visibility data suggest an emission region <38 uas in size, and at least two components, possibly polarized. We find a lower limit of the brightness temperature of the inner jet region of about 10^10 K. Lastly, we find an upper limit of 20% on the linear polarization fraction at a fringe spacing of ~38 uas. With APEX the angul...

  6. Diacylglycerol lipase a knockout mice demonstrate metabolic and behavioral phenotypes similar to those of cannabinoid receptor 1 knockout mice

    Directory of Open Access Journals (Sweden)

    David R Powell

    2015-06-01

    Full Text Available After creating >4650 knockouts (KOs of independent mouse genes, we screened them by high-throughput phenotyping and found that cannabinoid receptor 1 (Cnr1 KO mice had the same lean phenotype published by others. We asked if our KOs of DAG lipase a or b (Dagla or Daglb, which catalyze biosynthesis of the endocannabinoid (EC 2-Arachidonoylglycerol (2-AG, or Napepld, which catalyzes biosynthesis of the EC anandamide, shared the lean phenotype of Cnr1 KO mice. We found that Dagla KO mice, but not Daglb or Napepld KO mice, were among the leanest of 3651 chow-fed KO lines screened. In confirmatory studies, chow- or high fat diet-fed Dagla and Cnr1 KO mice were leaner than wild type (WT littermates; when data from multiple cohorts of adult mice were combined, body fat was 47% and 45% lower in Dagla and Cnr1 KO mice, respectively, relative to WT values. In contrast, neither Daglb nor Napepld KO mice were lean. Weanling Dagla KO mice ate less than WT mice and had body weight similar to pair-fed WT mice, and adult Dagla KO mice had normal activity and VO2 levels, similar to Cnr1 KO mice. Our Dagla and Cnr1 KO mice also had low fasting insulin, triglyceride and total cholesterol levels, and after a glucose challenge had normal glucose but very low insulin levels. Dagla and Cnr1 KO mice also showed similar responses to a battery of behavioral tests. These data suggest: 1 the lean phenotype of young Dagla and Cnr1 KO mice is mainly due to hypophagia; 2 in pathways where ECs signal through Cnr1 to regulate food intake and other metabolic and behavioral phenotypes observed in Cnr1 KO mice, Dagla alone provides the 2-AG that serves as the EC signal; and 3 small molecule Dagla inhibitors with a pharmacokinetic profile similar to that of Cnr1 inverse agonists are likely to mirror the ability of these Cnr1 inverse agonists to lower body weight and improve glycemic control in obese patients with type 2 diabetes, but may also induce undesirable neuropsychiatric

  7. Dentin Dysplasia in Notum Knockout Mice.

    Science.gov (United States)

    Vogel, P; Read, R W; Hansen, G M; Powell, D R; Kantaputra, P N; Zambrowicz, B; Brommage, R

    2016-07-01

    Secreted WNT proteins control cell differentiation and proliferation in many tissues, and NOTUM is a secreted enzyme that modulates WNT morphogens by removing a palmitoleoylate moiety that is essential for their activity. To better understand the role this enzyme in development, the authors produced NOTUM-deficient mice by targeted insertional disruption of the Notum gene. The authors discovered a critical role for NOTUM in dentin morphogenesis suggesting that increased WNT activity can disrupt odontoblast differentiation and orientation in both incisor and molar teeth. Although molars in Notum(-/-) mice had normal-shaped crowns and normal mantle dentin, the defective crown dentin resulted in enamel prone to fracture during mastication and made teeth more susceptible to endodontal inflammation and necrosis. The dentin dysplasia and short roots contributed to tooth hypermobility and to the spread of periodontal inflammation, which often progressed to periapical abscess formation. The additional incidental finding of renal agenesis in some Notum (-/-) mice indicated that NOTUM also has a role in kidney development, with undiagnosed bilateral renal agenesis most likely responsible for the observed decreased perinatal viability of Notum(-/-) mice. The findings support a significant role for NOTUM in modulating WNT signaling pathways that have pleiotropic effects on tooth and kidney development. PMID:26926082

  8. Necroptotic Cell Death Signaling and Execution Pathway: Lessons from Knockout Mice

    Directory of Open Access Journals (Sweden)

    José Belizário

    2015-01-01

    Full Text Available Under stress conditions, cells in living tissue die by apoptosis or necrosis depending on the activation of the key molecules within a dying cell that either transduce cell survival or death signals that actively destroy the sentenced cell. Multiple extracellular (pH, heat, oxidants, and detergents or intracellular (DNA damage and Ca2+ overload stress conditions trigger various types of the nuclear, endoplasmic reticulum (ER, cytoplasmatic, and mitochondrion-centered signaling events that allow cells to preserve the DNA integrity, protein folding, energetic, ionic and redox homeostasis, thus escaping from injury. Along the transition from reversible to irreversible injury, death signaling is highly heterogeneous and damaged cells may engage autophagy, apoptotic, or necrotic cell death programs. Studies on multiple double- and triple- knockout mice identified caspase-8, flip, and fadd genes as key regulators of embryonic lethality and inflammation. Caspase-8 has a critical role in pro- and antinecrotic signaling pathways leading to the activation of receptor interacting protein kinase 1 (RIPK1, RIPK3, and the mixed kinase domain-like (MLKL for a convergent execution pathway of necroptosis or regulated necrosis. Here we outline the recent discoveries into how the necrotic cell death execution pathway is engaged in many physiological and pathological outcome based on genetic analysis of knockout mice.

  9. Myeloid Deletion of α1AMPK Exacerbates Atherosclerosis in LDL Receptor Knockout (LDLRKO) Mice.

    Science.gov (United States)

    Cao, Qiang; Cui, Xin; Wu, Rui; Zha, Lin; Wang, Xianfeng; Parks, John S; Yu, Liqing; Shi, Hang; Xue, Bingzhong

    2016-06-01

    Macrophage inflammation marks all stages of atherogenesis, and AMPK is a regulator of macrophage inflammation. We therefore generated myeloid α1AMPK knockout (MAKO) mice on the LDL receptor knockout (LDLRKO) background to investigate whether myeloid deletion of α1AMPK exacerbates atherosclerosis. When fed an atherogenic diet, MAKO/LDLRKO mice displayed exacerbated atherosclerosis compared with LDLRKO mice. To determine the underlying pathophysiological pathways, we characterized macrophage inflammation/chemotaxis and lipid/cholesterol metabolism in MAKO/LDLRKO mice. Myeloid deletion of α1AMPK increased macrophage inflammatory gene expression and enhanced macrophage migration and adhesion to endothelial cells. Remarkably, MAKO/LDLRKO mice also displayed higher composition of circulating chemotaxically active Ly-6C(high) monocytes, enhanced atherosclerotic plaque chemokine expression, and monocyte recruitment into plaques, leading to increased atherosclerotic plaque macrophage content and inflammation. MAKO/LDLRKO mice also exhibited higher plasma LDL and VLDL cholesterol content, increased circulating apolipoprotein B (apoB) levels, and higher liver apoB expression. We conclude that macrophage α1AMPK deficiency promotes atherogenesis in LDLRKO mice and is associated with enhanced macrophage inflammation and hypercholesterolemia and that macrophage α1AMPK may serve as a therapeutic target for prevention and treatment of atherosclerosis. PMID:26822081

  10. Adipose-specific knockout of SEIPIN/BSCL2 results in progressive lipodystrophy.

    Science.gov (United States)

    Liu, Lu; Jiang, Qingqing; Wang, Xuhong; Zhang, Yuxi; Lin, Ruby C Y; Lam, Sin Man; Shui, Guanghou; Zhou, Linkang; Li, Peng; Wang, Yuhui; Cui, Xin; Gao, Mingming; Zhang, Ling; Lv, Ying; Xu, Guoheng; Liu, George; Zhao, Dong; Yang, Hongyuan

    2014-07-01

    Berardinelli-Seip congenital lipodystrophy type 2 (BSCL2) is the most severe form of human lipodystrophy, characterized by an almost complete loss of adipose tissue and severe insulin resistance. BSCL2 is caused by loss-of-function mutations in the BSCL2/SEIPIN gene, which is upregulated during adipogenesis and abundantly expressed in the adipose tissue. The physiological function of SEIPIN in mature adipocytes, however, remains to be elucidated. Here, we generated adipose-specific Seipin knockout (ASKO) mice, which exhibit adipocyte hypertrophy with enlarged lipid droplets, reduced lipolysis, adipose tissue inflammation, progressive loss of white and brown adipose tissue, insulin resistance, and hepatic steatosis. Lipidomic and microarray analyses revealed accumulation/imbalance of lipid species, including ceramides, in ASKO adipose tissue as well as increased endoplasmic reticulum stress. Interestingly, the ASKO mice almost completely phenocopy the fat-specific peroxisome proliferator-activated receptor-γ (Pparγ) knockout (FKO-γ) mice. Rosiglitazone treatment significantly improved a number of metabolic parameters of the ASKO mice, including insulin sensitivity. Our results therefore demonstrate a critical role of SEIPIN in maintaining lipid homeostasis and function of adipocytes and reveal an intimate relationship between SEIPIN and PPAR-γ.

  11. MAO A knockout attenuates adrenocortical response to various kinds of stress.

    Science.gov (United States)

    Popova, Nina K; Maslova, Larissa N; Morosova, Ekaterina A; Bulygina, Veta V; Seif, Isabelle

    2006-02-01

    The effect of a lack of the gene encoding monoamine oxidase A (MAO A) in transgenic Tg 8 mice on the corticosterone response to restraint, cold, water deprivation-induced, or social acute stress as well as chronic variable stress was studied. It was found that Tg 8 mice with genetic MAO A knockout and wild-type C3H/HeJ (C3H) strain showed similar plasma corticosterone resting level. MAO A knockout mice differed from C3H mice by attenuated response to restraint (60 min), cold (4 degrees C, 60 min), and water deprivation (48 h) as well as to a chronic (15 days) variable stress. No difference between Tg 8 and C3H strains in the response to psychosocial stress (encounters for 30 min of six previously isolated mice) has been found. ACTH administration to dexamethasone-pretreated mice produced a similar corticosterone effect in Tg 8 and C3H mice, indicating that the decreased stress response in MAO A-deficient mice was due rather to the central mechanisms regulating stress-induced ACTH release than to adrenocortical responsiveness to ACTH.

  12. Comparative Evaluation of Electronic Apex Locators and Radiovisiography for Working Length Determination in Primary Teeth in vivo

    Science.gov (United States)

    Singh, Neerja; Rathore, Monika S; Tandon, Shobha; Rajkumar, Balakrishnan

    2016-01-01

    ABSTRACT Aims: The aim of this study was to evaluate the effectiveness of two different varieties of electronic apex locators and radiovisiography (RVG) for working length determination in primary teeth. Materials and methods: A total of 30 primary teeth indicated for pulpectomy in children aged 3 to 8 years were randomly selected and subjected to working length determination using two varieties of electronic apex locators and RVG separately. The data were then subjected to statistical analysis. Results: A very strong correlation between electronic measurement methods and RVG length was observed. Conclusion: Radiovisiography and apex locators are equally effective in determining working length in primary teeth. How to cite this article: Abdullah A, Singh N, Rathore MS, Tandon S, Rajkumar B. Comparative Evaluation of Electronic Apex Locators and Radiovisiography for Working Length Determination in Primary Teeth in vivo. Int J Clin Pediatr Dent 2016;9(2):118-123. PMID:27365931

  13. New sub-millimeter heterodyne observations of CO and HCN in Titan's atmosphere with the APEX Swedish Heterodyne Facility Instrument

    CERN Document Server

    Rengel, M; Hartogh, P

    2010-01-01

    The origin of the atmosphere of the largest moon of Saturn, Titan, is poorly understood and its chemistry is rather complicated. Ground-based millimeter/sub-millimeter heterodyne spectroscopy resolves line shapes sufficiently to determine information in Titan's atmospheric composition (on vertical profiles and isotopic ratios). We test the capabilities of the Swedish Heterodyne Facility Instrument (SHFI), Receiver APEX-1, together with the Atacama Pathfinder EXperiment APEX 12-m telescope for Titan's atmospheric observations. In particular we present sub-millimeter observations of the CO(2-1) and HCN(3-2) lines of the Titan stratosphere with APEX, and with SHFI taken during the Science Verification (SV) instrument phase on March and June 2008. With the help of appropriate radiative transfer calculations we investigate the possibility to constrain the chemical concentrations and optimize the performance of the APEX-1 instrument for inferring vertical profiles of molecular components of the atmosphere of Titan.

  14. ATTENUATION OF DIFFRACTED MULTIPLES WITH AN APEX-SHIFTED TANGENT-SQUARED RADON TRANSFORM IN IMAGE SPACE

    Directory of Open Access Journals (Sweden)

    Alvarez Gabriel

    2006-12-01

    Full Text Available In this paper, we propose a method to attenuate diffracted multiples with an apex-shifted tangent-squared Radon transform in angle domain common image gathers (ADCIG . Usually, where diffracted multiples are a problem, the wave field propagation is complex and the moveout of primaries and multiples in data space is irregular. The method handles the complexity of the wave field propagation by wave-equation migration provided that migration velocities are reasonably accurate. As a result, the moveout of the multiples is well behaved in the ADCIGs. For 2D data, the apex-shifted tangent-squared Radon transform maps the 2D space image into a 3D space-cube model whose dimensions are depth, curvature and apex-shift distance.
    Well-corrected primaries map to or near the zero curvature plane and specularly-reflected multiples map to or near the zero apex-shift plane. Diffracted multiples map elsewhere in the cube according to their curvature and apex-shift distance. Thus, specularly reflected as well as diffracted multiples can be attenuated simultaneously. This approach is illustrated with a segment of a 2D seismic line over a large salt body in the Gulf of Mexico. It is shown that ignoring the apex shift compromises the attenuation of the diffracted multiples, whereas the approach proposed attenuates both the specularly-reflected and the diffracted multiples without compromising the primaries.

  15. Effects of lithium chloride on open-field behaviors in fragile X mental retardation 1 gene knockout mice%氯化锂改善脆性X染色体智力低下基因1敲除小鼠的旷场行为

    Institute of Scientific and Technical Information of China (English)

    张伟雯; 孙卫文; 杨泉; 黄月玲; 戴丽军; 杨凤啸; 陈盛强; 孙祺章

    2014-01-01

    化锂可以改善FMR1基因敲除小鼠的旷场异常行为,其机制与氯化锂导致的P-GSK3β的表达增加有关.%Objective To study the effects and potential mechanisms of lithium chloride on openfield behaviors of fragile X mental retardation 1 gene (FMR1) knockout mice (KO mice).Methods 4-weekold KO mice (n=90) and their wild type counterparts (WT mice,n=90) were,by using random number table,assigned to control group (intraperitoneally injected with normal saline,n=15) and 5 treatment groups intraperitoneally injected with 30 mg/kg (n=15),60 mg/kg (n=15),90 mg/kg (n=15),120 mg/kg (n=15) and 200 mg/kg lithium chloride (n=15) for 5 consecutive days,respectively.The differences in the total length of running,total number of region crossings,and the duration and number of central region crossings of all groups were determined in open field tests.Meanwhile,Western blotting was used to observe the expression of glycogen synthase kinase3β (GSK3β) and phosphorylated glycogen synthase kinase3β (PGSK3β) in the hippocampus and cortex of KO and WT mice.Results KO mice had longer distance of movements,greater total number of region crossings,longer duration and greater number of central region crossings than WT mice (all P<0.05).Compared with control group,administration of lithium chloride resulted in significantly reduced distance of movements,total number of region crossings,shorter duration and smaller number of central region crossings (all P<0.05) in KO mice.Lithium chloride administration (90,120 and 200 mg/kg) was associated with shorter distance of movements and duration of central region crossings in WT mice as compared with control group (all P<0.05).200 mg/kg,but not 30 to 120 mg/kg,lithium chloride led to reduced total number of region crossings compared with control group (75.73±5.12 vs 125.73±9.24,P<0.05).However,lithium chloride administration was associated with a reduced number of central region crossings compared with control group (all P<0.05).No

  16. Knockout driven reactions in complex molecules and their clusters

    Science.gov (United States)

    Gatchell, Michael; Zettergren, Henning

    2016-08-01

    Energetic ions lose some of their kinetic energy when interacting with electrons or nuclei in matter. Here, we discuss combined experimental and theoretical studies on such impulse driven reactions in polycyclic aromatic hydrocarbons (PAHs), fullerenes, and pure or mixed clusters of these molecules. These studies show that the nature of excitation is important for how complex molecular systems respond to ion/atom impact. Rutherford-like nuclear scattering processes may lead to prompt atom knockout and formation of highly reactive fragments, while heating of the molecular electron clouds in general lead to formation of more stable and less reactive fragments. In this topical review, we focus on recent studies of knockout driven reactions, and present new calculations of the angular dependent threshold (displacement) energies for such processes in PAHs. The so-formed fragments may efficiently form covalent bonds with neighboring molecules in clusters. These unique molecular growth processes may be important in astrophysical environments such as low velocity shock waves.

  17. Neutron knockout in neutral-current neutrino-oxygen interactions

    CERN Document Server

    Ankowski, Artur M

    2013-01-01

    The ongoing and future searches for diffuse supernova neutrinos and sterile neutrinos carried out with large water-Cherenkov detectors require a precise determination of the backgrounds, especially those involving gamma rays. Of great importance, in this context, is the process of neutron knockout through neutral-current (NC) scattering of atmospheric neutrinos on oxygen. Nuclear reinteractions of the produced neutron may in fact lead to the production of gamma rays of energies high enough to mimic the processes of interest. In this Letter, we focus on the kinematical range suitable for simulations of atmospheric-neutrino interactions and provide the neutron-knockout cross sections computed using the formalism based on realistic nuclear spectral function. The role of the strange-quark contribution to the NC axial form factor is also analyzed. Based on the available experimental information, we give an estimate of the associated uncertainty.

  18. A Conditional Knockout Mouse Line of the Oxytocin Receptor

    OpenAIRE

    Lee, Heon-Jin; Heather K Caldwell; Macbeth, Abbe H.; Tolu, Selen G.; Young, W. Scott

    2008-01-01

    Oxytocin plays important roles in reproductive physiology and various behaviors, including maternal behavior and social memory. Its receptor (Oxtr) is present in peripheral tissues and brain, so a conditional knockout (KO, −/−) would be useful to allow elimination of the receptor in specific sites at defined times. We created a line of mice in which loxP sites flank Oxtr coding sequence (floxed) enable Cre recombinase-mediated inactivation of the receptor. We expressed Cre recombinase in thes...

  19. One-neutron knockout from {sup 51-55}Sc

    Energy Technology Data Exchange (ETDEWEB)

    Schwertel, S.; Maierbeck, P.; Gernhaeuser, R.; Bildstein, V.; Boehmer, M.; Eppinger, K.; Faestermann, T.; Friese, J.; Fabbietti, L.; Maier, L.; Winkler, S. [Technische Universitaet Muenchen, Physik Department E12, Garching (Germany); Kruecken, R. [Technische Universitaet Muenchen, Physik Department E12, Garching (Germany); TRIUMF, Vancouver (Canada); University of British Columbia, Department of Physics and Astronomy, Vancouver (Canada); Kroell, T. [Technische Universitaet Muenchen, Physik Department E12, Garching (Germany); Technische Universitaet Darmstadt, Institut fuer Kernphysik, Darmstadt (Germany); Alvarez-Pol, H.; Benjamim, E.A.; Benlliure, J.; Caamano, M.; Cortina-Gil, D.; Gascon, M.; Kurtukian, T.; Perez, D.; Rodriguez-Tajes, C. [Universidade de Santiago de Compostela, Departamento de Fisica de Particulas, Santiago de Compostela (Spain); Aksouh, F.; Aumann, T.; Behr, K.; Boretzky, K.; Bruenle, A.; Chatillon, A.; Chulkov, L.V.; Geissel, H.; Gerl, J.; Gorska, M.; Kojouharov, I.; Klimkiewicz, A.; Kurz, N.; Nociforo, C.; Schaffner, H.; Simon, H.; Stanoiu, M.; Suemmerer, K.; Weick, H. [GSI Helmholtzzentrum fuer Schwerionenforschung, Darmstadt (Germany); Borge, M.J.G.; Pascual-Izarra, C.; Perea, A.; Tengblad, O. [CSIC, Instituto de Estructura de la Materia, Madrid (Spain); Buerger, A. [University of Oslo, SAFE/OCL, Oslo (Norway); CEA, Gif-sur-Yvette (France); Casarejos, E.; Brown, B.A. [University of Vigo, Vigo (Spain); Enders, J.; Schrieder, G. [Technische Universitaet Darmstadt, Institut fuer Kernphysik, Darmstadt (Germany); Hansen, P.G. [Michigan State University, NSCL, East Lansing, Michigan (United States); Jonson, B.; Nyman, G. [Chalmers Tekniska Hoegskola och Goeteborgs Universitet, Experimentell Fysik, Goeteborg (Sweden); Kanungo, R. [TRIUMF, Vancouver (Canada); GSI Helmholtzzentrum fuer Schwerionenforschung, Darmstadt (Germany); Saint Mary' s University, Halifax (Canada); Kiselev, O. [GSI Helmholtzzentrum fuer Schwerionenforschung, Darmstadt (Germany); Johannes Gutenberg Universitaet, Mainz (Germany); Paul Scherrer Institut, Villigen (Switzerland); Larsson, K. [GSI Helmholtzzentrum fuer Schwerionenforschung, Darmstadt (Germany); Chalmers Tekniska Hoegskola och Goeteborgs Universitet, Experimentell Fysik, Goeteborg (Sweden); Le Bleis, T. [GSI Helmholtzzentrum fuer Schwerionenforschung, Darmstadt (Germany); IN2P3-CNRS/Universite Louis Pasteur, Institut Pluridisciplinaire Hubert Curien, Strasbourg Cedex 2 (France); Mahata, K. [GSI Helmholtzzentrum fuer Schwerionenforschung, Darmstadt (Germany); Paul Scherrer Institut, Villigen (Switzerland); Nilsson, T. [Technische Universitaet Darmstadt, Institut fuer Kernphysik, Darmstadt (Germany); Chalmers Tekniska Hoegskola och Goeteborgs Universitet, Experimentell Fysik, Goeteborg (Sweden); Prochazka, A. [GSI Helmholtzzentrum fuer Schwerionenforschung, Darmstadt (Germany); Comenius University, Faculty of Mathematics and Physics, Bratislava (Slovakia); Rossi, D. [Johannes Gutenberg Universitaet, Mainz (Germany); Sitar, B. [Comenius University, Faculty of Mathematics and Physics, Bratislava (Slovakia); Otsuka, T. [University of Tokyo, Hongo, Bunkyo-ku, Department of Physics, Tokyo (Japan); Tostevin, J.A. [University of Surrey, Department of Physics, Faculty of Engineering and Physical Sciences, Guildford (United Kingdom); Rae, W.D.M. [Garsington, Oxfordshire (United Kingdom)

    2012-12-15

    Results are presented from a one-neutron knockout experiment at relativistic energies of {approx} 420 A MeV on {sup 51-55}Sc using the GSI Fragment Separator as a two-stage magnetic spectrometer and the MINIBALL array for gamma-ray detection. Inclusive longitudinal momentum distributions and cross-sections were measured enabling the determination of the contributions corresponding to knockout from the {nu}p{sub 1/2}, {nu}p{sub 3/2}, (L = 1) and {nu}f{sub 7/2}, {nu}f{sub 5/2} (L = 3) neutron orbitals. The observed L = 1 and L = 3 contributions are compared with theoretical cross-sections using eikonal knockout theory and spectroscopic factors from shell model calculations using the GXPF1A interaction. The measured inclusive knockout cross-sections generally follow the trends expected theoretically and given by the spectroscopic strength predicted from the shell model calculations. However, the deduced L = 1 cross-sections are generally 30-40% higher while the L = 3 contributions are about a factor of two smaller than predicted. This points to a promotion of neutrons from the {nu}f{sub 7/2} to the {nu}p{sub 3/2} orbital indicating a weakening of the N = 28 shell gap in these nuclei. While this is not predicted for the phenomenological GXPF1A interaction such a weakening is predicted by recent calculations using realistic low-momentum interactions V{sub low} {sub k} obtained by evolving a chiral N3LO nucleon-nucleon potential. (orig.)

  20. Impaired Conditioned Taste Aversion Learning in Spinophilin Knockout Mice

    OpenAIRE

    Stafstrom-Davis, Carrie A.; Ouimet, Charles C.; Feng, Jian; Allen, Patrick B; Greengard, Paul; Houpt, Thomas A.

    2001-01-01

    Plasticity in dendritic spines may underlie learning and memory. Spinophilin, a protein enriched in dendritic spines, has the properties of a scaffolding protein and is believed to regulate actin cytoskeletal dynamics affecting dendritic spine morphology. It also binds protein phosphatase-1 (PP-1), an enzyme that regulates dendritic spine physiology. In this study, we tested the role of spinophilin in conditioned taste aversion learning (CTA) using transgenic spinophilin knockout mice. CTA is...

  1. Lessons from hepatocyte-specific cyp51 knockout mice

    OpenAIRE

    Keber, Rok; Lorbek, Gregor; Lewinska, Monika; Juvan, Peter; Perše, Martina; Bjorkhem, Ingemar; Rozman, Damjana; Horvat, Simon; Jeruc, Jera; Gutiérrez Mariscal, Francisco Miguel; Gebhardt, Rolf

    2016-01-01

    We demonstrate unequivocally that defective cholesterol synthesis is an independent determinant of liver inflammation and fibrosis. We prepared a mouse hepatocyte-specific knockout (LKO) of lanosterol 14 a -demethylase (CYP51) from the part of cholesterol synthesis that is already committed to cholesterol. LKO mice developed hepatomegaly with oval cell proliferation, fibrosis and inflammation, but without steatosis. The key trigger was reduced cholesterol esters that provoked cell cycle arres...

  2. Screening Methods to Identify TALEN-Mediated Knockout Mice

    OpenAIRE

    Nakagawa, Yoshiko; Yamamoto, Takashi; Suzuki, Ken-Ichi; Araki, Kimi; Takeda, Naoki; Ohmuraya, Masaki; Sakuma, Tetsushi

    2014-01-01

    Genome editing with site-specific nucleases, such as zinc-finger nucleases or transcription activator-like effector nucleases (TALENs), and RNA-guided nucleases, such as the CRISPR/Cas (clustered regularly interspaced short palindromic repeats/CRISPR-associated) system, is becoming the new standard for targeted genome modification in various organisms. Application of these techniques to the manufacture of knockout mice would be greatly aided by simple and easy methods for genotyping of mutant...

  3. CRISPR-Mediated Triple Knockout of SLAMF1, SLAMF5 and SLAMF6 Supports Positive Signaling Roles in NKT Cell Development

    Science.gov (United States)

    Huang, Bonnie; Gomez-Rodriguez, Julio; Preite, Silvia; Garrett, Lisa J.; Harper, Ursula L.; Schwartzberg, Pamela L.

    2016-01-01

    The SLAM family receptors contribute to diverse aspects of lymphocyte biology and signal via the small adaptor molecule SAP. Mutations affecting SAP lead to X-linked lymphoproliferative syndrome Type 1, a severe immunodysregulation characterized by fulminant mononucleosis, dysgammaglobulinemia, and lymphoproliferation/lymphomas. Patients and mice having mutations affecting SAP also lack germinal centers due to a defect in T:B cell interactions and are devoid of invariant NKT (iNKT) cells. However, which and how SLAM family members contribute to these phenotypes remains uncertain. Three SLAM family members: SLAMF1, SLAMF5 and SLAMF6, are highly expressed on T follicular helper cells and germinal center B cells. SLAMF1 and SLAMF6 are also implicated in iNKT development. Although individual receptor knockout mice have limited iNKT and germinal center phenotypes compared to SAP knockout mice, the generation of multi-receptor knockout mice has been challenging, due to the genomic linkage of the genes encoding SLAM family members. Here, we used Cas9/CRISPR-based mutagenesis to generate mutations simultaneously in Slamf1, Slamf5 and Slamf6. Genetic disruption of all three receptors in triple-knockout mice (TKO) did not grossly affect conventional T or B cell development and led to mild defects in germinal center formation post-immunization. However, the TKO worsened defects in iNKT cells development seen in SLAMF6 single gene-targeted mice, supporting data on positive signaling and potential redundancy between these receptors. PMID:27258160

  4. Role of the Molybdoflavoenzyme Aldehyde Oxidase Homolog 2 in the Biosynthesis of Retinoic Acid: Generation and Characterization of a Knockout Mouse▿ †

    Science.gov (United States)

    Terao, Mineko; Kurosaki, Mami; Barzago, Maria Monica; Fratelli, Maddalena; Bagnati, Renzo; Bastone, Antonio; Giudice, Chiara; Scanziani, Eugenio; Mancuso, Alessandra; Tiveron, Cecilia; Garattini, Enrico

    2009-01-01

    The mouse aldehyde oxidase AOH2 (aldehyde oxidase homolog 2) is a molybdoflavoenzyme. Harderian glands are the richest source of AOH2, although the protein is detectable also in sebaceous glands, epidermis, and other keratinized epithelia. The levels of AOH2 in the Harderian gland and skin are controlled by genetic background, being maximal in CD1 and C57BL/6 and minimal in DBA/2, CBA, and 129/Sv strains. Testosterone is a negative regulator of AOH2 in Harderian glands. Purified AOH2 oxidizes retinaldehyde into retinoic acid, while it is devoid of pyridoxal-oxidizing activity. Aoh2−/− mice, the first aldehyde oxidase knockout animals ever generated, are viable and fertile. The data obtained for this knockout model indicate a significant role of AOH2 in the local synthesis and biodisposition of endogenous retinoids in the Harderian gland and skin. The Harderian gland's transcriptome of knockout mice demonstrates overall downregulation of direct retinoid-dependent genes as well as perturbations in pathways controlling lipid homeostasis and cellular secretion, particularly in sexually immature animals. The skin of knockout mice is characterized by thickening of the epidermis in basal conditions and after UV light exposure. This has correlates in the corresponding transcriptome, which shows enrichment and overall upregulation of genes involved in hypertrophic responses. PMID:18981221

  5. A satellite cell-specific knockout of the androgen receptor reveals myostatin as a direct androgen target in skeletal muscle.

    Science.gov (United States)

    Dubois, Vanessa; Laurent, Michaël R; Sinnesael, Mieke; Cielen, Nele; Helsen, Christine; Clinckemalie, Liesbeth; Spans, Lien; Gayan-Ramirez, Ghislaine; Deldicque, Louise; Hespel, Peter; Carmeliet, Geert; Vanderschueren, Dirk; Claessens, Frank

    2014-07-01

    Androgens have well-established anabolic actions on skeletal muscle, although the direct effects of the androgen receptor (AR) in muscle remain unclear. We generated satellite cell-specific AR-knockout (satARKO) mice in which the AR is selectively ablated in satellite cells, the muscle precursor cells. Total-limb maximal grip strength is decreased by 7% in satARKO mice, with soleus muscles containing ∼10% more type I fibers and 10% less type IIa fibers than the corresponding control littermates. The weight of the perineal levator ani muscle is markedly reduced (-52%). Thus, muscle AR is involved in fiber-type distribution and force production of the limb muscles, while it is a major determinant of the perineal muscle mass. Surprisingly, myostatin (Mstn), a strong inhibitor of skeletal muscle growth, is one of the most androgen-responsive genes (6-fold reduction in satARKO) through direct transcription activation by the AR. Consequently, muscle hypertrophy in response to androgens is augmented in Mstn-knockout mice. Our finding that androgens induce Mstn signaling to restrain their own anabolic actions has implications for the treatment of muscle wasting disorders.-Dubois, V., Laurent, M. R., Sinnesael, M., Cielen, N., Helsen, C., Clinckemalie, L., Spans, L., Gayan-Ramirez, G., Deldicque, L., Hespel, P., Carmeliet, G., Vanderschueren, D., and Claessens, F. A satellite cell-specific knockout of the androgen receptor reveals myostatin as a direct androgen target in skeletal muscle.

  6. Failed stabilization for long-term potentiation in the auditory cortex of FMR1 knockout mice.

    Directory of Open Access Journals (Sweden)

    Sungchil Yang

    Full Text Available Fragile X syndrome is a developmental disorder that affects sensory systems. A null mutation of the Fragile X Mental Retardation protein 1 (Fmr1 gene in mice has varied effects on developmental plasticity in different sensory systems, including normal barrel cortical plasticity, altered ocular dominance plasticity and grossly impaired auditory frequency map plasticity. The mutation also has different effects on long-term synaptic plasticity in somatosensory and visual cortical neurons, providing insights on how it may differentially affect the sensory systems. Here we present evidence that long-term potentiation (LTP is impaired in the developing auditory cortex of the Fmr1 knockout (KO mice. This impairment of synaptic plasticity is consistent with impaired frequency map plasticity in the Fmr1 KO mouse. Together, these results suggest a potential role of LTP in sensory map plasticity during early sensory development.

  7. Selective Deactivation of Gibberellins below the Shoot Apex is Critical to Flowering but Not to Stem Elongation of Lolium

    DEFF Research Database (Denmark)

    King, Rod W; Mander, Lewis N; Asp, Torben;

    2008-01-01

    in their effectiveness for flowering because they are deactivated by C-2 hydroxylation below the shoot apex. In contrast, GA5 is effective because of its structural protection at C-2. Excised vegetative shoot tips rapidly degrade [14C]GA1, [14C]GA4, and [14C]GA20 (>80% in 6 h), but not [14C]GA5. Coincidentally, genes...... in florally induced shoot tips and these GAs can become active for floral development. Structural changes which stabilize GA4 confirm the link between florigenicity and restricted GA 2β-hydroxylation (e.g. 2 -hydroxylation and C-2 di-methylation). Additionally, a 2-oxidase inhibitor (Trinexapac Ethyl...

  8. Gene

    Data.gov (United States)

    U.S. Department of Health & Human Services — Gene integrates information from a wide range of species. A record may include nomenclature, Reference Sequences (RefSeqs), maps, pathways, variations, phenotypes,...

  9. Pregnane X receptor knockout mice display aging-dependent wearing of articular cartilage.

    Directory of Open Access Journals (Sweden)

    Kotaro Azuma

    Full Text Available Steroid and xenobiotic receptor (SXR and its murine ortholog, pregnane X receptor (PXR, are nuclear receptors that are expressed at high levels in the liver and the intestine where they function as xenobiotic sensors that induce expression of genes involved in detoxification and drug excretion. Recent evidence showed that SXR and PXR are also expressed in bone tissue where they mediate bone metabolism. Here we report that systemic deletion of PXR results in aging-dependent wearing of articular cartilage of knee joints. Histomorphometrical analysis showed remarkable reduction of width and an enlarged gap between femoral and tibial articular cartilage in PXR knockout mice. We hypothesized that genes induced by SXR in chondrocytes have a protective effect on articular cartilage and identified Fam20a (family with sequence similarity 20a as an SXR-dependent gene induced by the known SXR ligands, rifampicin and vitamin K2. Lastly, we demonstrated the biological significance of Fam20a expression in chondrocytes by evaluating osteoarthritis-related gene expression of primary articular chondrocytes. Consistent with epidemiological findings, our results indicate that SXR/PXR protects against aging-dependent wearing of articular cartilage and that ligands for SXR/PXR have potential role in preventing osteoarthritis caused by aging.

  10. Does murine spermatogenesis require WNT signalling? A lesson from Gpr177 conditional knockout mouse models.

    Science.gov (United States)

    Chen, Su-Ren; Tang, J-X; Cheng, J-M; Hao, X-X; Wang, Y-Q; Wang, X-X; Liu, Y-X

    2016-01-01

    Wingless-related MMTV integration site (WNT) proteins and several other components of the WNT signalling pathway are expressed in the murine testes. However, mice mutant for WNT signalling effector β-catenin using different Cre drivers have phenotypes that are inconsistent with each other. The complexity and overlapping expression of WNT signalling cascades have prevented researchers from dissecting their function in spermatogenesis. Depletion of the Gpr177 gene (the mouse orthologue of Drosophila Wntless), which is required for the secretion of various WNTs, makes it possible to genetically dissect the overall effect of WNTs in testis development. In this study, the Gpr177 gene was conditionally depleted in germ cells (Gpr177(flox/flox), Mvh-Cre; Gpr177(flox/flox), Stra8-Cre) and Sertoli cells (Gpr177(flox/flox), Amh-Cre). No obvious defects in fertility and spermatogenesis were observed in these three Gpr177 conditional knockout (cKO) mice at 8 weeks. However, late-onset testicular atrophy and fertility decline in two germ cell-specific Gpr177 deletion mice were noted at 8 months. In contrast, we did not observe any abnormalities of spermatogenesis and fertility, even in 8-month-old Gpr177(flox/flox), Amh-Cre mice. Elevation of reactive oxygen species (ROS) was detected in Gpr177 cKO germ cells and Sertoli cells and exhibited an age-dependent manner. However, significant increase in the activity of Caspase 3 was only observed in germ cells from 8-month-old germ cell-specific Gpr177 knockout mice. In conclusion, GPR177 in Sertoli cells had no apparent influence on spermatogenesis, whereas loss of GPR177 in germ cells disrupted spermatogenesis in an age-dependent manner via elevating ROS levels and triggering germ cell apoptosis. PMID:27362799

  11. Curative high dose rate vaginal apex brachytherapy in stage I papillary serous carcinoma of the endometrium

    International Nuclear Information System (INIS)

    Introduction: Uterine papillary serous carcinoma (UPSC) is a morphologically distinct variant of endometrial carcinoma that is associated with a poor prognosis, high recurrence rate, clinical understaging, and poor response to salvage treatment. We describe the presentation, local and distant control, survival, salvage rate, and complications for patients undergoing whole abdominal radiation therapy (WART), low dose rate (LDR) intracavitary brachytherapy, or high dose rate (HDR) vaginal brachytherapy in patients with stage I UPSC. Methods: Between 1976 and 1994 more than 1700 patients with endometrial carcinoma were treated with radiation therapy, 30 patients with stage I UPSC (1.8%) were treated with radiation before or following TAH/BSO. All patients underwent either preoperative Simon's packing or tandem and plaque which delivered 30-40 Gy to the serosa, WART, or HDR Ir-192 vaginal apex brachytherapy to a total dose of 21 Gy in 3 fractions at 0.5 cm from the vaginal mucosa. A total of 14 patients received HDR vaginal brachytherapy and (5(14)) patients received systemic chemotherapy. All patients presented with vaginal bleeding at a median age of 67 years (range 34-88). The group of 30 patients underwent TAH/BSO, 17 patients were completely staged pathologically (pelvic and para-aortic lymph nodes, omentectomy, and pelvic washings), and 2 patients underwent omental biopsy and pelvic washings only. All specimens revealed UPSC, nuclear grade 3, and lymphovascular invasion (23%). The pathologic stage was IA: 23% (7), IB: 67% (20), and IC: 10% (3). The median follow-up for all patients was 49 months (range 13-187 months). For the patients receiving postoperative HDR vaginal brachytherapy the median time from surgery to radiation was 42 days (range 29-91). Results: The 5-year actuarial disease free survival for Figo stage I UPSC patients treated with postoperative HDR vaginal brachytherapy and systemic chemotherapy was 100% compared to 74% for stage I UPSC patient

  12. HISTOLOGICINVESTIGATION OF PULPAL INFLAMMATION AND NECROSIS ON TRUMATIZED OPEN APEX CAT CANINE TEETH

    Directory of Open Access Journals (Sweden)

    AA KHADEMI

    2002-09-01

    Full Text Available Interoduction. Treatment of trumatized crown fractured teeth with open apex is a major problem in endodontic treatment. Long term stability of such teeth depends upon the maintainace of pulp vitality. The time period between the accident and the treatment affects the pulp vitality as well as the type of treatment and prognosis. The objective of this study was to provide a rough time table of pulpal inflammation and necrosis after the exposure of crow - fractured teeth to oral Environment. Methods. Fourty eight canine teeth in 12 young cats were used in this experiment. To make sure the teeth apecies were all open, they were checked by radiograph. The crown of all the teeth were cut 2mm from the CEJ and the pulps were left open to oral inviornment for periods of 1, 3, 7, 14, 28, 90 days. Animals were then sacrificed by vital perfusion fixation technique. Results. The mean depth of pulpal inflammation and necrosis for periods of 1, 3, 7, 14, 28 and 90 days were 1.98, 3.3, 2.39, 2.84, 3.92 and 8.57, respectively. There was no statistically significant difference between the time periods of 1, 7 and 14 days. Where as, time periods of 14 and 28 days showed significant difference. In 90 day time period, all the teeth showed total pulpal necrosis and was significantly different from all the other groups. Discussion. It appears that, trumatized open apex teeth exposed to oral Enviornment for less than 28 days could be well treated by apexogenesis. Considering the sufficient blood supply to open apex teeth in cases of deep inflammation and necrosis the teeth could be saved by partial pulpectomy. Only cases of total pulp necrosis should be treated by apexification technique.

  13. Calibration, Sensor Model Improvements and Uncertainty Budget of the Airborne Imaging Spectrometer APEX

    Science.gov (United States)

    Hueni, A.

    2015-12-01

    ESA's Airborne Imaging Spectrometer APEX (Airborne Prism Experiment) was developed under the PRODEX (PROgramme de Développement d'EXpériences scientifiques) program by a Swiss-Belgian consortium and entered its operational phase at the end of 2010 (Schaepman et al., 2015). Work on the sensor model has been carried out extensively within the framework of European Metrology Research Program as part of the Metrology for Earth Observation and Climate (MetEOC and MetEOC2). The focus has been to improve laboratory calibration procedures in order to reduce uncertainties, to establish a laboratory uncertainty budget and to upgrade the sensor model to compensate for sensor specific biases. The updated sensor model relies largely on data collected during dedicated characterisation experiments in the APEX calibration home base but includes airborne data as well where the simulation of environmental conditions in the given laboratory setup was not feasible. The additions to the model deal with artefacts caused by environmental changes and electronic features, namely the impact of ambient air pressure changes on the radiometry in combination with dichroic coatings, influences of external air temperatures and consequently instrument baffle temperatures on the radiometry, and electronic anomalies causing radiometric errors in the four shortwave infrared detector readout blocks. Many of these resolved issues might be expected to be present in other imaging spectrometers to some degree or in some variation. Consequently, the work clearly shows the difficulties of extending a laboratory-based uncertainty to data collected under in-flight conditions. The results are hence not only of interest to the calibration scientist but also to the spectroscopy end user, in particular when commercial sensor systems are used for data collection and relevant sensor characteristic information tends to be sparse. Schaepman, et al, 2015. Advanced radiometry measurements and Earth science

  14. Apex-to-Cupola Distance Following VATS Predicts Recurrence in Patients With Primary Spontaneous Pneumothorax.

    Science.gov (United States)

    Chang, Jia-Ming; Lai, Wu-Wei; Yen, Yi-Ting; Tseng, Yau-Lin; Chen, Ying-Yuan; Wu, Ming-Ho; Chen, Wei; Light, Richard W

    2015-09-01

    Our study sought to determine whether the size of the residual apical pleural space in young patients with primary spontaneous pneumothorax (PSP) following video-assisted thoracoscopic surgery is associated with the risk of recurrence. We retrospectively reviewed patients (≤30 years' old) with primary spontaneous pneumothorax following thoracoscopic surgery (2002-2010) in a university-affiliated hospital. The size of residual apical pleural space was estimated by measuring the apex-to-cupola distance on a postoperative chest radiograph at 2 time windows: first between postoperative day (POD) 0 and 3, and second between POD 4 and 14. A total of 149 patients were enrolled with a median follow-up of 11.2 months (interquartile range, 0.95-29.5 months), of whom 141 (94.6%) were male with a mean age of 20 years. The postoperative recurrence rate was 11.4%. Comparing the characteristics between the patients with and without recurrent pneumothorax, the patients with recurrence were younger (18.2 + 2.4 vs 20.7 + 3.7 years, P = 0.008), with a lower rate of pleurodesis (35% vs1 69%, P = 0.037), longer apex-to-cupola distance at POD 0 to 3 (22.41 ± 19.56 vs 10.07 ± 10.83 mm, P pneumothorax, age 10 mm (P = 0.027, OR: 5.319), and no pleurodesis during VATS (P = 0.022, OR: 5.042) were independent risk factors for recurrent pneumothorax. The recurrence rate was not low (11.4%) in young patients with PSP following VATS. Residual apical pleural space with apex-to-cupola distance of 10 mm or greater at POD 0 to 3, younger age, and no pleurodesis would increase postoperative recurrence of primary spontaneous pneumothorax.

  15. Lake bathymetry and species occurrence predict the distribution of a lacustrine apex predator.

    Science.gov (United States)

    Hughes, M R; Dodd, J A; Maitland, P S; Adams, C E

    2016-04-01

    This study examined the abiotic and biotic characteristics of ecosystems that allow expression of a life history called ferox trout, the colloquial name given to brown trout Salmo trutta adopting a piscivorous life history strategy, an apex predator in post-glacial lakes in northern Europe. One hundred and ninety-two lakes in Scotland show evidence of currently, or historically, supporting ferox S. trutta; their presence was predicted in logistic models by larger and deeper lakes with a large catchment that also support populations of Arctic charr Salvelinus alpinus. PMID:26899559

  16. A COMPARISON BETWEEN SOUNDING DATA AND WRF FORECASTS AT APEX SITE

    OpenAIRE

    M. Caneo; D. Pozo; L. Illanes; Curé, M.

    2011-01-01

    Cinco configuraciones de WRF usando diferentes modelos de suelo y parametrizaciones de microfísica y de capa límite planetaria se evaluaron con sondeos lanzados durante una campaña de mediciones en el sitio de APEX (Atacama Pathfinder EXperiment). Los resultados indican que los cambios en la parametrización de microfísica no producen cambios apreciables en los perfiles de humedad. El modelo de suelo de Noah muestra menores errores en los perfiles verticales de las variables analizadas en comp...

  17. SEU results from the Advanced Photovoltaic and Electronics Experiments (APEX) satellite

    Energy Technology Data Exchange (ETDEWEB)

    Mullen, E.G.; Ray, K.P. [Phillips Lab., Hanscom AFB, MA (United States); Koga, R. [Aerospace Corp., Los Angeles, CA (United States); Holeman, E.G.; Delorey, D.E. [Boston Coll., Newton, MA (United States). Inst. for Space Research

    1995-12-01

    The APEX satellite, launched in August of 1994, had a solid state data recorder (SSDR) as its onboard data storage system. The recorder contained 220 4Mbit X 1 Hitachi DRAMs of which 176 were routinely interrogated for SEUs and corrected with an EDAC code. Corrections were recorded in the spacecraft housekeeping files and are the basis of this study. The SEU rates and spatial locations are compared to in-situ particle measurements and to ground test results from devices from the same lot flown. The results show that properly designed SSDRs are a viable alternative to conventional tap recording systems for all orbits in near-Earth space.

  18. Aged neuronal nitric oxide knockout mice show preserved olfactory learning in both social recognition and odor-conditioning tasks

    OpenAIRE

    Keith Maurice Kendrick

    2015-01-01

    There is evidence for both neurotoxic and neuroprotective roles of nitric oxide (NO) in the brain and changes in the expression of the neuronal isoform of nitric oxide synthase (nNOS) gene occur during aging. The current studies have investigated potential support for either a neurotoxic or neuroprotective role of NO derived from nNOS in the context of aging by comparing olfactory learning and locomotor function in young compared to old nNOS knockout (nNOS/-) and wildtype control mice. Tasks ...

  19. Aged neuronal nitric oxide knockout mice show preserved olfactory learning in both social recognition and odor-conditioning tasks

    OpenAIRE

    James, Bronwen M.; Li, Qin; Luo, Lizhu; Kendrick, Keith M.

    2015-01-01

    There is evidence for both neurotoxic and neuroprotective roles of nitric oxide (NO) in the brain and changes in the expression of the neuronal isoform of NO synthase (nNOS) gene occur during aging. The current studies have investigated potential support for either a neurotoxic or neuroprotective role of NO derived from nNOS in the context of aging by comparing olfactory learning and locomotor function in young compared to old nNOS knockout (nNOS−/−) and wildtype control mice. Tasks involving...

  20. Quantification of oxidative stress phenotypes based on high-throughput growth profiling of protein kinase and phosphatase knockouts

    DEFF Research Database (Denmark)

    Altıntaş, Ali; Martini, Jacopo; Mortensen, Uffe Hasbro;

    2016-01-01

    Cellular responses to oxidative stress are important for restoring redox balance and ensuring cell survival. Genetic defects in response factors can lead to impaired response to oxidative damage and contribute to disease and aging. In single cell organisms, such as yeasts, the integrity of the ox...... able to identify surprisingly diverse sensitivity and resistance patterns as a function of gene knockout.......Cellular responses to oxidative stress are important for restoring redox balance and ensuring cell survival. Genetic defects in response factors can lead to impaired response to oxidative damage and contribute to disease and aging. In single cell organisms, such as yeasts, the integrity...

  1. U19/Eaf2 knockout causes lung adenocarcinoma, B-cell lymphoma, hepatocellular carcinoma and prostatic intraepithelial neoplasia

    OpenAIRE

    Xiao, W; Zhang, Q; Habermacher, G; Yang, X.; Zhang, A-y; Cai, X.; Hahn, J.; Liu, J; Pins, M; Doglio, L; Dhir, R; Gingrich, J; Wang, Z.(Institute of High Energy Physics, Beijing, China)

    2007-01-01

    Upregulated gene 19 (U19)/ELL-associated factor 2 (Eaf2) is a potential human tumor suppressor that exhibits frequent allelic loss and downregulation in high-grade prostate cancer. U19/Eaf2, along with its homolog Eaf1, has been reported to regulate transcriptional elongation via interaction with the eleven-nineteen lysine-rich leukemia (ELL) family of proteins. To further explore the tumor-suppressive effects of U19/Eaf2, we constructed and characterized a murine U19/Eaf2-knockout model. Hom...

  2. Male germ cell-specific knockout of cholesterogenic cytochrome P450 lanosterol 14α-demethylase (Cyp51)[S

    OpenAIRE

    Keber, Rok; Ačimovič, Jure; Majdič, Gregor; Motaln, Helena; Rozman, Damjana; Horvat, Simon

    2013-01-01

    Cytochrome P450 lanosterol 14α-demethylase (CYP51) and its products, meiosis-activating sterols (MASs), were hypothesized by previous in vitro studies to have an important role in regulating meiosis and reproduction. To test this in vivo, we generated a conditional male germ cell-specific knockout of the gene Cyp51 in the mouse. High excision efficiency of Cyp51 allele in germ cells resulted in 85–89% downregulation of Cyp51 mRNA and protein levels in germ cells. Quantitative metabolic profil...

  3. Selective increase of dark phase water intake in neuropeptide-Y Y2 and Y4 receptor knockout mice

    OpenAIRE

    Wultsch, Thomas; Painsipp, Evelin; Donner, Sabine; Sperk1, Günther; Herzog, Herbert; Peskar, Bernhard A; Holzer, Peter

    2005-01-01

    Neuropeptide-Y (NPY) is involved in the regulation of ingestive behaviour and energy homeostasis. Since deletion of the NPY Y2 and Y4 receptor gene increases and decreases food intake, respectively, we examined whether water intake during the light and dark phase is altered in Y2 and Y4 receptor knockout mice. The water consumption of mice staying in their home cages was measured by weighing the water bottles at the beginning and end of the light phase during 4 consecutive days. Control, Y2 a...

  4. Optical properties of aerosols during APEX and ACE-Asia experiments

    Science.gov (United States)

    Sano, Itaru; Mukai, Sonoyo; Okada, Yasuhiko; Holben, Brent N.; Ohta, Sachio; Takamura, Tamio

    2003-12-01

    Sun/sky photometry and polarimetry of atmospheric light have been undertaken by multispectral photometers (CE-318-1 and -2, Cimel Electronique, France) and a polarimeter (PSR-1000, Opto Research, Japan) over Amami, Noto, and Shirahama, Japan, during APEX-E1, -E2, and ACE-Asia field campaigns. Radiometers provide us with the optical thickness of aerosols and Ångström exponent. Other aerosol characteristics, e.g., size distribution, refractive index, etc., are retrieved based on each inversion method corresponding each equipment. The former takes a standard AERONET processing, and the latter is according to our own procedure to analyze the polarimetry with PSR-1000. After several aerosol parameters are derived, the HYSPLIT4 backward trajectory analysis is adopted to search the origin of aerosols. It is shown from these ground measurements that aerosol optical thickness, Ångström exponent, and refractive index are classified into two typical categories as a background type detected in winter, and a soil dust type appeared in Asian dust events in spring. Further, it is found that the obtained size distribution of Asian dust indicates the dominance of large particles.

  5. The mass distribution of clumps within infrared dark clouds. A Large APEX Bolometer Camera study

    CERN Document Server

    Gomez, Laura; Schuller, Frederic; Menten, Karl; Ballesteros-Paredes, Javier

    2013-01-01

    We present an analysis of the dust continuum emission at 870 um in order to investigate the mass distribution of clumps within infrared dark clouds (IRDCs). We map six IRDCs with the Large APEX BOlometer CAmera (LABOCA) at APEX, reaching an rms noise level of 28-44 mJy/beam. The dust continuum emission coming from these IRDCs was decomposed by using two automated algorithms, Gaussclumps and Clumpfind. Moreover, we carried out single-pointing observations of the N_2H^+ (3-2) line toward selected positions to obtain kinematic information. The mapped IRDCs are located in the range of kinematic distances of 2.7-3.2 kpc. We identify 510 and 352 sources with Gaussclumps and Clumpfind, respectively, and estimate masses and other physical properties assuming a uniform dust temperature. The mass ranges are 6-2692 Msun (Gaussclumps) and 7-4254 Msun (Clumpfind) and the ranges in effective radius are around 0.10-0.74 pc (Gaussclumps) and 0.16-0.99 pc (Clumpfind). The mass distribution, independent of the decomposition me...

  6. Medullary Venous Hypertension Secondary to a Petrous Apex Dural Arteriovenous Fistula: A Case Report

    Directory of Open Access Journals (Sweden)

    Meghan Murphy

    2012-11-01

    Full Text Available Background: Dural arteriovenous fistulae (dAVF are common intracranial vascular lesions typically becoming symptomatic with cortical venous hypertension and possible hemorrhage. Here, we present a case illustration of a petrous apex dAVF with marked medullary venous hypertension and a unique clinical presentation. Methods: Case report. Results: A 72-year-old female, whose clinical progression was significant for altered mental status and progressive weakness, presented with diplopia, right leg paresis, and ataxia. Magnetic resonance imaging revealed edema involving the medulla. On digital subtraction cerebral angiogram, the patient was found to have a petrous apex dAVF, Cognard type IV. Following treatment with Onyx embolization, her symptoms rapidly improved, with complete resolution of diplopia and drastic improvement of her ataxia. Conclusion: The importance of this case is in the presentation and deterioration of the clinical exam, resembling an acute ischemic event. Further, this case illustrates that dAVF may cause venous hypertension with rapid onset of focal neurologic symptoms not exclusive to cortical locations.

  7. New electronic apex locator Romiapex A-15 presented accuracy for working length determination in permanent teeth

    Directory of Open Access Journals (Sweden)

    Etevaldo Matos Maia Filho

    2014-12-01

    Full Text Available Purpose: The present study aims to evaluate, ex vivo, the accuracy of electronic apex locators Root ZX II and Romiapex-15, for working length (WL determination in permanent teeth. Materials and Methods: Fourteen single-rooted teeth (incisors and canines, with their apices fully formed were used. The dental crowns were removed. The anatomic length of the tooth (real measurement was visually determined through the insertion of a size 10 K-file until the tip of the instrument could be observed in the apical foramen under a microscope (20X. Teeth were fixed in a model of resin and adapted into alginate soaked with saline solution, which was used as an  electrical conductor. Using a K-file, root canals were measured electronically using both devices. The results obtained for each apex locator were compared to the real measurements. The accuracy between the devices was statistically analyzed using the Bland-Altman graph, Intraclass Correlation Coefficient (ICC, and Student’s t-test. Results: The mean difference between the measurements using the Root ZX II was 0.277mm greater than the real measurement, while the measurements from the Romiapex-15 were 0.308mm higher on average. The comparison between Root ZX II and Romiapex-15 had no significant difference (p= 0.868. Conclusion: It was concluded that Root ZX II and Romiapex-15 had similar accuracy. Romiapex-15 could be an option for WL determination in permanent teeth.

  8. ATLASGAL - The APEX Telescope Large Area Survey of the Galaxy at 870 microns

    CERN Document Server

    Schuller, F; Contreras, Y; Wyrowski, F; Schilke, P; Bronfman, L; Henning, T; Walmsley, C M; Beuther, H; Bontemps, S; Cesaroni, R; Deharveng, L; Garay, G; Herpin, F; Le Floc'h, B; Linz, H; Mardones, D; Minier, V; Molinari, S; Motte, F; Nyman, L -A; Reveret, V; Risacher, C; Russeil, D; Schneider, N; Testi, L; Troost, T; Vasyunina, T; Wienen, M; Zavagno, A; Kovács, A; Kreysa, E; Siringo, G; Weiss, A

    2009-01-01

    (Abridged) Studying continuum emission from interstellar dust is essential to locate and characterize the highest density regions in the interstellar medium. In particular, the early stages of massive star formation are still mysterious. Our goal is to produce a large scale, systematic database of massive pre- and proto-stellar clumps in the Galaxy, in order to better understand how and under what conditions star formation takes place. A well characterized sample of star-forming sites will deliver an evolutionary sequence and a mass function of high-mass star-forming clumps. Such a systematic survey at submm wavelengths also represents a pioneering work in preparation for Herschel and ALMA. The APEX telescope is ideally located to observe the inner Milky Way. The recently commissioned Large APEX Bolometer Camera (LABOCA) is a 295-element bolometer array observing at 870 microns, with a beam of 19". Taking advantage of its large field of view (11.4') and excellent sensitivity, we have started an unbiased surve...

  9. In vitro study of apical leakage of root canals after different root apex resection techniques

    Directory of Open Access Journals (Sweden)

    André Augusto Franco MARQUES

    2008-12-01

    Full Text Available Objective:The aim of the present study was to evaluate in vitro apicalmarginal leakage of root canals after three root apex resectiontechniques.Material and methods: Thirty maxillary canines wereprepared chemo-mechanically to a size 40 master apical file, andenlarged by using Gates Glidden burs 2, 3 and 4. Teeth were obturatedby Tagger thermomechanical compaction technique and then dividedinto three groups (n = 10. The first group (GI was 3 mm apicalresected with Endo Z in high speed handpiece. The specimens of secondgroup (GII were 3 mm apical weared with Endo Z, and the third group(GIII had 3 mm apical resected with Er:YAG laser (500 impulse, 12pps and 300 mJ energy. After the apical sections, IRM cement wasplaced into the root-end cavity and teeth of each group were immersedin India ink for 5 days and submitted to decalcification and clarifying for marginal apical microleakage visualization with microscope.Results: The results showed statistically difference (p < 0.01 between GI and the other groups.Conclusion: The root apex cut using Endo Z promotes the lowest values of microleakage when used in apicoectomy.

  10. Actively produced high-energy electron bursts within the magnetosphere: the APEX project

    Directory of Open Access Journals (Sweden)

    L. Přech

    Full Text Available The APEX project (Active Plasma Experiment has been launched into a polar orbit in December 1991 and consists of two satellites (IK-25 and MAGION-3, with a distance between them from 200 km to 10 000 km. The mission used intensive electron beam emission, complemented by a low-energy Xenon plasma generator during the electron beam injection, for the study of dynamic processes in the magnetosphere and upper ionosphere. 

    The paper deals with short, intensive bursts of field-aligned electrons observed during the APEX mission on board the MAGION-3 satellite. These events are located pre-dominantly at the middle geomagnetic latitudes in the day-side magnetosphere. The time-energy structure of these electron bursts is similar to the inverted-V one, but the pitch-angle width is less than 10°. Electrons with an energy up to 700 keV are often observed during the events. We analyze the observed events, discuss the possible mechanisms of the particle spreading, and the role of the main satellite’s activity as a possible source of these events.

    Key words. Ionosphere (particle acceleration; particle precipitation – Space plasma physics (active perturbation experiments

  11. Effect of pre-flaring and file size on the accuracy of two electronic apex locators

    Directory of Open Access Journals (Sweden)

    Manoel Brito-Júnior

    2012-10-01

    Full Text Available OBJECTIVE: This ex vivo study evaluated the effect of pre-flaring and file size on the accuracy of the Root ZX and Novapex electronic apex locators (EALs. MATERIAL AND METHODS: The actual working length (WL was set 1 mm short of the apical foramen in the palatal root canals of 24 extracted maxillary molars. The teeth were embedded in an alginate mold, and two examiners performed the electronic measurements using #10, #15, and #20 K-files. The files were inserted into the root canals until the "0.0" or "APEX" signals were observed on the LED or display screens for the Novapex and Root ZX, respectively, retracting to the 1.0 mark. The measurements were repeated after the preflaring using the S1 and SX Pro-Taper instruments. Two measurements were performed for each condition and the means were used. Intra-class correlation coefficients (ICCs were calculated to verify the intra- and inter-examiner agreement. The mean differences between the WL and electronic length values were analyzed by the three-way ANOVA test (p0.8 and the results demonstrated a similar accuracy for both EALs (p>0.05. Statistically significant accurate measurements were verified in the pre-flared canals, except for the Novapex using a #20 K-file. CONCLUSIONS: The tested EALs showed acceptable accuracy, whereas the pre-flaring procedure revealed a more significant effect than the used file size.

  12. Utilising the resources of the International Knockout Mouse Consortium: the Australian experience.

    Science.gov (United States)

    Cotton, Leanne M; Meilak, Michelle L; Templeton, Tanya; Gonzales, Jose G; Nenci, Arianna; Cooney, Melissa; Truman, Dirk; Rodda, Fleur; Lynas, Alyce; Viney, Elizabeth; Rosenthal, Nadia; Bianco, Deborah M; O'Bryan, Moira K; Smyth, Ian M

    2015-04-01

    Mouse models play a key role in the understanding gene function, human development and disease. In 2007, the Australian Government provided funding to establish the Monash University embryonic stem cell-to-mouse (ES2M) facility. This was part of the broader Australian Phenomics Network, a national infrastructure initiative aimed at maximising access to global resources for understanding gene function in the mouse. The remit of the ES2M facility is to provide subsidised access for Australian biomedical researchers to the ES cell resources available from the International Knockout Mouse Consortium (IKMC). The stated aim of the IKMC is to generate a genetically modified mouse ES cell line for all of the ~23,000 genes in the mouse genome. The principal function of the Monash University ES2M service is to import genetically modified ES cells into Australia and to convert them into live mice with the potential to study human disease. Through advantages of economy of scale and established relationships with ES cell repositories worldwide, we have created over 110 germline mouse strains sourced from all of the major ES providers worldwide. We comment on our experience in generating these mouse lines; providing a snapshot of a "clients" perspective of using the IKMC resource and one which we hope will serve as a guide to other institutions or organisations contemplating establishing a similar centralised service. PMID:25645994

  13. CRISPR/Cas9-mediated GJA8 knockout in rabbits recapitulates human congenital cataracts.

    Science.gov (United States)

    Yuan, Lin; Sui, Tingting; Chen, Mao; Deng, Jichao; Huang, Yongye; Zeng, Jian; Lv, Qingyan; Song, Yuning; Li, Zhanjun; Lai, Liangxue

    2016-01-01

    Cataracts are the leading cause of vision loss in the world, although surgical treatment can restore vision in cataract patients. Until now, there have been no adequate animal models for in vivo studies of artificial lens safety and drug interactions. Genetic studies have demonstrated that GJA8 is involved in maintaining lens opacity and proper lens development. In this study, a cataract model with GJA8 gene knockout was developed via co-injection of Cas9/sgRNA mRNA into rabbit zygotes. Our results showed that gene mutation efficiency in the GJA8 locus reached 98.7% in embryos and 100% in pups, demonstrating that the Cas9/sgRNA system is a highly efficient tool for gene editing in rabbits. In agreement with other studies, our genetic and histology results showed that impaired GJA8 function caused microphthalmia, small lens size and cataracts. In summary, our novel rabbit model of cataracts will be an important drug-screening tool for cataract prevention and treatment. PMID:26912477

  14. Chronic minocycline treatment improves social recognition memory in adult male Fmr1 knockout mice.

    Science.gov (United States)

    Yau, Suk Yu; Chiu, Christine; Vetrici, Mariana; Christie, Brian R

    2016-10-01

    Fragile X syndrome (FXS) is caused by a mutation in the Fmr1 gene that leads to silencing of the gene and a loss of its gene product, Fragile X mental retardation protein (FMRP). Some of the key behavioral phenotypes for FXS include abnormal social anxiety and sociability. Here we show that Fmr1 knock-out (KO) mice exhibit impaired social recognition when presented with a novel mouse, and they display normal social interactions in other sociability tests. Administering minocycline to Fmr1 KO mice throughout critical stages of neural development improved social recognition memory in the novel mouse recognition task. To determine if synaptic changes in the prefrontal cortex (PFC) could have played a role in this improvement, we examined PSD-95, a member of the membrane-associated guanylate kinase family, and signaling molecules (ERK1/2, and Akt) linked to synaptic plasticity in the PFC. Our analyses indicated that while minocycline treatment can enhance behavioral performance, it does not enhance expression of PSD-95, ERK1/2 or Akt in the PFC. PMID:27291517

  15. Comprehensive reanalysis of transcription factor knockout expression data in Saccharomyces cerevisiae reveals many new targets.

    Science.gov (United States)

    Reimand, Jüri; Vaquerizas, Juan M; Todd, Annabel E; Vilo, Jaak; Luscombe, Nicholas M

    2010-08-01

    Transcription factor (TF) perturbation experiments give valuable insights into gene regulation. Genome-scale evidence from microarray measurements may be used to identify regulatory interactions between TFs and targets. Recently, Hu and colleagues published a comprehensive study covering 269 TF knockout mutants for the yeast Saccharomyces cerevisiae. However, the information that can be extracted from this valuable dataset is limited by the method employed to process the microarray data. Here, we present a reanalysis of the original data using improved statistical techniques freely available from the BioConductor project. We identify over 100,000 differentially expressed genes-nine times the total reported by Hu et al. We validate the biological significance of these genes by assessing their functions, the occurrence of upstream TF-binding sites, and the prevalence of protein-protein interactions. The reanalysed dataset outperforms the original across all measures, indicating that we have uncovered a vastly expanded list of relevant targets. In summary, this work presents a high-quality reanalysis that maximizes the information contained in the Hu et al. compendium. The dataset is available from ArrayExpress (accession: E-MTAB-109) and it will be invaluable to any scientist interested in the yeast transcriptional regulatory system. PMID:20385592

  16. Helicobacter pylori arginase mutant colonizes arginase Ⅱ knockout mice

    Institute of Scientific and Technical Information of China (English)

    Songhee H Kim; Melanie L Langford; Jean-Luc Boucher; Traci L Testerman; David J McGee

    2011-01-01

    AIM: To investigate the role of host and bacterial argi-nases in the colonization of mice by Helicobacter pylori (H. Pylori).METHODS: H. Pylori produces a very powerful urease that hydrolyzes urea to carbon dioxide and ammonium, which neutralizes acid. Urease is absolutely essential to H. Pylori pathogenesis; therefore, the urea substrate must be in ample supply for urease to work efficiently. The urea substrate is most likely provided by arginase activity, which hydrolyzes L-arginine to L-ornithine and urea. Previous work has demonstrated that H. Pylori arginase is surprisingly not required for colonization of wild-type mice. Hence, another in vivo source of the critical urea substrate must exist. We hypothesized that the urea source was provided by host arginase Ⅱ, since this enzyme is expressed in the stomach, and H. Pylori has previously been shown to induce the expres-sion of murine gastric arginase Ⅱ. To test this hypoth-esis, wild-type and arginase (rocF) mutant H. Pylori strain SS1 were inoculated into arginase Ⅱ knockout mice. RESULTS: Surprisingly, both the wild-type and rocF mutant bacteria still colonized arginase Ⅱ knock-out mice. Moreover, feeding arginase Ⅱ knockout mice the host arginase inhibitor S-(2-boronoethyl)-L-cysteine (BEC), while inhibiting > 50% of the host arginase Ⅰactivity in several tissues, did not block the ability of the rocF mutant H. Pylori to colonize. In con-trast, BEC poorly inhibited H. Pylori arginase activity. CONCLUSION: The in vivo source for the essential urea utilized by H. Pylori urease is neither bacterial arginase nor host arginase Ⅱ; instead, either residual host arginase Ⅰor agmatinase is probably responsible.

  17. 柴胡桂枝汤挥发油对脆性X综合征基因敲除小鼠旷场行为和氨基酸递质影响%Influence of Buplcurl and Ramulj Cinnamomi Decoction on levels of amino acid and beharious in open field in FMRI gene knockout mice

    Institute of Scientific and Technical Information of China (English)

    黄庆辉; 高飞; 刘忠民; 张华宋; 沈岩松; 陈盛强

    2012-01-01

    Objective To explore the effect of Bupleuri and Ramuli Cinnamomi Decoction essential oil in open fiel test and its relationship to level of amino acid. Methods FVB strain four-week mice, including FMR1 knockout (KO) and wild type (WT) counterparts were used. They were intraperitoneal injected by different concentrations Bupleuri and Ramuli Cinnamomi Decoction to observe open field behavior indicators and establish an effective drug concentration curve.and the mouse brain tissues were taken to make 1:10 (weight and volume ratio)tissue homogenate to determine level of amino acid. Results Compared with blank groups,the average speed, total distance and times of going through fields reduced in treatment groups, The level of γ-aminobutyric acid and as-partate in WT blank group was obviously lower than WT control mice, while the level of glutamate in WT blank group was higher than WT control group. Otherwise, the level of glutamate in WT blank group was higher than WT treated group (P<0.05). Conclusion Bupleuri and Ramuli Cinnamomi Decoction essential oil inhibits exploratory, excitability and motility in Fmrl knockout mice. The effect of Bupleuri and Ramuli Cinnamomi Decoction may be related to reduced ratio of excitatory amino acid / inhibitory amino acids.%目的 探讨柴胡桂枝汤挥发油对脆性X综合征基因敲除小鼠旷场行为和氨基酸递质的影响.方法 采用不同浓度柴胡桂枝汤挥发油对脆性X综合征基因敲除小鼠小鼠进行处理,分析了柴胡桂枝汤挥发油对小鼠旷场行为和氨基酸递质的影响.结果 未用药的WT组和KO组小鼠比较,KO组小鼠的运动性、兴奋性、探索性明显高于WT组小鼠.随着柴胡桂枝汤挥发油浓度的增加,KO和WT小鼠的平均速度、运动轨迹及穿越中央格次数均逐渐降低.WT空白组小鼠脑组织中γ-氨基丁酸水平和甘氨酸明显降低,而谷氨酸水平则明显升高,此差异有统计学意义:与此同时wt空白组小鼠与WT用药

  18. Zinc finger nuclease mediated knockout of ADP-dependent glucokinase in cancer cell lines: effects on cell survival and mitochondrial oxidative metabolism.

    Directory of Open Access Journals (Sweden)

    Susan Richter

    Full Text Available Zinc finger nucleases (ZFN are powerful tools for editing genes in cells. Here we use ZFNs to interrogate the biological function of ADPGK, which encodes an ADP-dependent glucokinase (ADPGK, in human tumour cell lines. The hypothesis we tested is that ADPGK utilises ADP to phosphorylate glucose under conditions where ATP becomes limiting, such as hypoxia. We characterised two ZFN knockout clones in each of two lines (H460 and HCT116. All four clones had frameshift mutations in all alleles at the target site in exon 1 of ADPGK, and were ADPGK-null by immunoblotting. ADPGK knockout had little or no effect on cell proliferation, but compromised the ability of H460 cells to survive siRNA silencing of hexokinase-2 under oxic conditions, with clonogenic survival falling from 21±3% for the parental line to 6.4±0.8% (p = 0.002 and 4.3±0.8% (p = 0.001 for the two knockouts. A similar increased sensitivity to clonogenic cell killing was observed under anoxia. No such changes were found when ADPGK was knocked out in HCT116 cells, for which the parental line was less sensitive than H460 to anoxia and to hexokinase-2 silencing. While knockout of ADPGK in HCT116 cells caused few changes in global gene expression, knockout of ADPGK in H460 cells caused notable up-regulation of mRNAs encoding cell adhesion proteins. Surprisingly, we could discern no consistent effect on glycolysis as measured by glucose consumption or lactate formation under anoxia, or extracellular acidification rate (Seahorse XF analyser under oxic conditions in a variety of media. However, oxygen consumption rates were generally lower in the ADPGK knockouts, in some cases markedly so. Collectively, the results demonstrate that ADPGK can contribute to tumour cell survival under conditions of high glycolytic dependence, but the phenotype resulting from knockout of ADPGK is cell line dependent and appears to be unrelated to priming of glycolysis in these lines.

  19. Zinc Finger Nuclease Mediated Knockout of ADP-Dependent Glucokinase in Cancer Cell Lines: Effects on Cell Survival and Mitochondrial Oxidative Metabolism

    Science.gov (United States)

    Richter, Susan; Morrison, Shona; Connor, Tim; Su, Jiechuang; Print, Cristin G.; Ronimus, Ron S.; McGee, Sean L.; Wilson, William R.

    2013-01-01

    Zinc finger nucleases (ZFN) are powerful tools for editing genes in cells. Here we use ZFNs to interrogate the biological function of ADPGK, which encodes an ADP-dependent glucokinase (ADPGK), in human tumour cell lines. The hypothesis we tested is that ADPGK utilises ADP to phosphorylate glucose under conditions where ATP becomes limiting, such as hypoxia. We characterised two ZFN knockout clones in each of two lines (H460 and HCT116). All four clones had frameshift mutations in all alleles at the target site in exon 1 of ADPGK, and were ADPGK-null by immunoblotting. ADPGK knockout had little or no effect on cell proliferation, but compromised the ability of H460 cells to survive siRNA silencing of hexokinase-2 under oxic conditions, with clonogenic survival falling from 21±3% for the parental line to 6.4±0.8% (p = 0.002) and 4.3±0.8% (p = 0.001) for the two knockouts. A similar increased sensitivity to clonogenic cell killing was observed under anoxia. No such changes were found when ADPGK was knocked out in HCT116 cells, for which the parental line was less sensitive than H460 to anoxia and to hexokinase-2 silencing. While knockout of ADPGK in HCT116 cells caused few changes in global gene expression, knockout of ADPGK in H460 cells caused notable up-regulation of mRNAs encoding cell adhesion proteins. Surprisingly, we could discern no consistent effect on glycolysis as measured by glucose consumption or lactate formation under anoxia, or extracellular acidification rate (Seahorse XF analyser) under oxic conditions in a variety of media. However, oxygen consumption rates were generally lower in the ADPGK knockouts, in some cases markedly so. Collectively, the results demonstrate that ADPGK can contribute to tumour cell survival under conditions of high glycolytic dependence, but the phenotype resulting from knockout of ADPGK is cell line dependent and appears to be unrelated to priming of glycolysis in these lines. PMID:23799003

  20. Expression of PPARα modifies fatty acid effects on insulin secretion in uncoupling protein-2 knockout mice

    Directory of Open Access Journals (Sweden)

    Chan Catherine B

    2007-03-01

    Full Text Available Abstract Aims/hypothesis In uncoupling protein-2 (UCP2 knockout (KO mice, protection of beta cells from fatty acid exposure is dependent upon transcriptional events mediated by peroxisome proliferator-activated receptor-α (PPARα. Methods PPARα expression was reduced in isolated islets from UCP2KO and wild-type (WT mice with siRNA for PPARα (siPPARα overnight. Some islets were also cultured with oleic or palmitic acid, then glucose stimulated insulin secretion (GSIS was measured. Expression of genes was examined by quantitative RT-PCR or immunoblotting. PPARα activation was assessed by oligonucleotide consensus sequence binding. Results siPPARα treatment reduced PPARα protein expression in KO and WT islets by >85%. In siPPARα-treated UCP2KO islets, PA but not OA treatment significantly decreased the insulin response to 16.5 mM glucose. In WT islets, siPPARα treatment did not modify GSIS in PA and OA exposed groups. In WT islets, PA treatment significantly increased UCP2 mRNA and protein expression. Both PA and OA treatment significantly increased PPARα expression in UCP2KO and WT islets but OA treatment augmented PPARα protein expression only in UCP2KO islets (p Conclusion These data show that the negative effect of saturated fatty acid on GSIS is mediated by PPARα/UCP2. Knockout of UCP2 protects beta-cells from PA exposure. However, in the absence of both UCP2 and PPARα even a short exposure (24 h to PA significantly impairs GSIS.

  1. p21{sup WAF1/Cip1/Sdi1} knockout mice respond to doxorubicin with reduced cardiotoxicity

    Energy Technology Data Exchange (ETDEWEB)

    Terrand, Jerome; Xu, Beibei; Morrissy, Steve; Dinh, Thai Nho [Department of Pharmacology,College of Medicine, University of Arizona, 1501 N. Campbell Ave, Tucson, AZ 85724 (United States); Williams, Stuart [Biomedical Engineering Program, College of Medicine, University of Arizona, 1501 N. Campbell Ave, Tucson, AZ 85724 (United States); Chen, Qin M., E-mail: qchen@email.arizona.edu [Department of Pharmacology,College of Medicine, University of Arizona, 1501 N. Campbell Ave, Tucson, AZ 85724 (United States)

    2011-11-15

    Doxorubicin (Dox) is an antineoplastic agent that can cause cardiomyopathy in humans and experimental animals. As an inducer of reactive oxygen species and a DNA damaging agent, Dox causes elevated expression of p21{sup WAF1/Cip1/Sdi1} (p21) gene. Elevated levels of p21 mRNA and p21 protein have been detected in the myocardium of mice following Dox treatment. With chronic treatment of Dox, wild type (WT) animals develop cardiomyopathy evidenced by elongated nuclei, mitochondrial swelling, myofilamental disarray, reduced cardiac output, reduced ejection fraction, reduced left ventricular contractility, and elevated expression of ANF gene. In contrast, p21 knockout (p21KO) mice did not show significant changes in the same parameters in response to Dox treatment. In an effort to understand the mechanism of the resistance against Dox induced cardiomyopathy, we measured levels of antioxidant enzymes and found that p21KO mice did not contain elevated basal or inducible levels of glutathione peroxidase and catalase. Measurements of 6 circulating cytokines indicated elevation of IL-6, IL-12, IFN{gamma} and TNF{alpha} in Dox treated WT mice but not p21KO mice. Dox induced elevation of IL-6 mRNA was detected in the myocardium of WT mice but not p21KO mice. While the mechanism of the resistance against Dox induced cardiomyopathy remains unclear, lack of inflammatory response may contribute to the observed cardiac protection in p21KO mice. -- Highlights: Black-Right-Pointing-Pointer Doxorubicin induces p21 elevation in the myocardium. Black-Right-Pointing-Pointer Doxorubicin causes dilated cardiomyopathy in wild type mice. Black-Right-Pointing-Pointer p21 Knockout mice are resistant against doxorubicin induced cardiomyopathy. Black-Right-Pointing-Pointer Lack of inflammatory response correlates with the resistance in p21 knockout mice.

  2. Auxin/AID versus conventional knockouts: distinguishing the roles of CENP-T/W in mitotic kinetochore assembly and stability

    Science.gov (United States)

    Wood, Laura; Booth, Daniel G.; Vargiu, Giulia; Ohta, Shinya; deLima Alves, Flavia; Samejima, Kumiko; Fukagawa, Tatsuo; Rappsilber, Juri; Earnshaw, William C.

    2016-01-01

    Most studies using knockout technologies to examine protein function have relied either on shutting off transcription (conventional conditional knockouts with tetracycline-regulated gene expression or gene disruption) or destroying the mature mRNA (RNAi technology). In both cases, the target protein is lost at a rate determined by its intrinsic half-life. Thus, protein levels typically fall over at least 1–3 days, and cells continue to cycle while exposed to a decreasing concentration of the protein. Here we characterise the kinetochore proteome of mitotic chromosomes isolated from a cell line in which the essential kinetochore protein CENP-T is present as an auxin-inducible degron (AID) fusion protein that is fully functional and able to support the viability of the cells. Stripping of the protein from chromosomes in early mitosis via targeted proteasomal degradation reveals the dependency of other proteins on CENP-T for their maintenance in kinetochores. We compare these results with the kinetochore proteome of conventional CENP-T/W knockouts. As the cell cycle is mostly formed from G1, S and G2 phases a gradual loss of CENP-T/W levels is more likely to reflect dependencies associated with kinetochore assembly pre-mitosis and upon entry into mitosis. Interestingly, a putative super-complex involving Rod-Zw10-zwilch (RZZ complex), Spindly, Mad1/Mad2 and CENP-E requires the function of CENP-T/W during kinetochore assembly for its stable association with the outer kinetochore, but once assembled remains associated with chromosomes after stripping of CENP-T during mitosis. This study highlights the different roles core kinetochore components may play in the assembly of kinetochores (upon entry into mitosis) versus the maintenance of specific components (during mitosis). PMID:26791246

  3. Normal Maternal Behavior, But Increased Pup Mortality, in Conditional Oxytocin Receptor Knockout Females

    OpenAIRE

    Macbeth, Abbe H.; Stepp, Jennifer E.; Lee, Heon-Jin; Young, W. Scott; Heather K Caldwell

    2010-01-01

    Oxytocin (Oxt) and the Oxt receptor (Oxtr) are implicated in the onset of maternal behavior in a variety of species. Recently, we developed two Oxtr knockout lines: a total body knockout (Oxtr−/−) and a conditional Oxtr knockout (OxtrFB/FB) in which the Oxtr is lacking only in regions of the forebrain, allowing knockout females to potentially nurse and care for their biological offspring. In the current study, we assessed maternal behavior of postpartum OxtrFB/FB females toward their own pups...

  4. Knockout reaction induced by 6He at 61.2 MeV/u

    Institute of Scientific and Technical Information of China (English)

    LU Lin-Hui; CAO Zhong-Xin; SONG Yu-Shou; XIAO Jun; LI Qi-Te; QIAO Rui; YOU Hai-Bo; CHEN Rui-Jiu; XU Hu-Shan; WANG Jian-Song; GUO Zhong-Yan; YE Yan-Lin; ZHANG Xue-Ying; LI Chen; HU Zheng-Guo; CHEN Ruo-FU; WANG Meng; XU Zhi-Guo; YUE Ke; TANG Bin; ZANG Yong-Dong; ZHANG Xue-Heng; JIANG Dong-Xing; YAO Xiang-Wu; CHEN Jin-Da; BAI Zhen; HUA Hui; ZHENG Tao; LI Zhi-Huan; GE Yu-Cheng; LI Xiang-Qing; LOU Jian-Ling

    2011-01-01

    A knockout reaction induced by 6He at 61.2 MeV/u was carried out at the HIRFL-RIBLL radioactive beam line.The α core fragments at forward angles were detected in coincidence with the recoiled protons at large angles.From this coincident measurement the valence nucleon knockout mechanism and the core knockout mechanism can be separated according to the polar angle correlation between the core fragments and the recoiled protons.It is demonstrated that,when reconstructing the resonant state of a weakly bound nucleus,the contamination resulting from the core knockout mechanism should be eliminated in order to obtain the correct structure information.

  5. A Rare Case of Congenital Heart Disease with Bifid Car- diac Apex: A Unique Finding in Left Ventricle

    Directory of Open Access Journals (Sweden)

    Majid Maleki

    2015-10-01

    Full Text Available Bifid cardiac apex is a rare anomaly of human hearts. We report of the case of a 34-year-old man with a previous history of ventricular septal defect (VSD and subvalvular pulmonary stenosis. He had undergone pulmonary commissurotomy and VSD closure 22 years before he was referred to our center for evaluation of progressive dyspnea. Transthoracic echocardiography revealed atrial septal defect (ASD, multiple VSDs, severe pulmonary regurgitation, and a bifid cardiac apex. The patient was referred for re-do surgery for ASD and VSD closure along with pulmonary valve replacement, but he refused the surgery.

  6. Quantum mechanical understanding of field dependence of the apex barrier of a single-wall carbon nanotube

    Science.gov (United States)

    Peng, Jie; Li, Zhibing; He, Chunshan; Deng, Shaozhi; Xu, Ningsheng; Zheng, Xiao; Chen, Guanhua

    2005-12-01

    The potential barrier at the apex of a single-wall carbon nanotube emitter is found to be strongly and nonlinearly dependent on the external applied field, due to a quantum mechanical mechanism instead of the correction of image potential in Fowler-Nordheim theory. The field enhancement factor depends on the applied field and is much smaller than that predicted by the classical theory. The field induced apex-vacuum barrier lowering is confirmed to be the essential mechanism for efficient field electron emission from capped carbon nanotubes.

  7. A STAT-1 knockout mouse model for Machupo virus pathogenesis

    Directory of Open Access Journals (Sweden)

    Shurtleff Amy C

    2011-06-01

    Full Text Available Abstract Background Machupo virus (MACV, a member of the Arenaviridae, causes Bolivian hemorrhagic fever, with ~20% lethality in humans. The pathogenesis of MACV infection is poorly understood, and there are no clinically proven treatments for disease. This is due, in part, to a paucity of small animal models for MACV infection in which to discover and explore candidate therapeutics. Methods Mice lacking signal transducer and activator of transcription 1 (STAT-1 were infected with MACV. Lethality, viral replication, metabolic changes, hematology, histopathology, and systemic cytokine expression were analyzed throughout the course of infection. Results We report here that STAT-1 knockout mice succumbed to MACV infection within 7-8 days, and presented some relevant clinical and histopathological manifestations of disease. Furthermore, the model was used to validate the efficacy of ribavirin in protection against infection. Conclusions The STAT-1 knockout mouse model can be a useful small animal model for drug testing and preliminary immunological analysis of lethal MACV infection.

  8. APEX Snaps First Close-up of Star Factories in Distant Universe

    Science.gov (United States)

    2010-03-01

    For the first time, astronomers have made direct measurements of the size and brightness of regions of star-birth in a very distant galaxy, thanks to a chance discovery with the APEX telescope. The galaxy is so distant, and its light has taken so long to reach us, that we see it as it was 10 billion years ago. A cosmic "gravitational lens" is magnifying the galaxy, giving us a close-up view that would otherwise be impossible. This lucky break reveals a hectic and vigorous star-forming life for galaxies in the early Universe, with stellar nurseries forming one hundred times faster than in more recent galaxies. The research is published online today in the journal Nature. Astronomers were observing a massive galaxy cluster [1] with the Atacama Pathfinder Experiment (APEX) telescope, using submillimetre wavelengths of light, when they found a new and uniquely bright galaxy, more distant than the cluster and the brightest very distant galaxy ever seen at submillimetre wavelengths. It is so bright because the cosmic dust grains in the galaxy are glowing after being heated by starlight. The new galaxy has been given the name SMM J2135-0102. "We were stunned to find a surprisingly bright object that wasn't at the expected position. We soon realised it was a previously unknown and more distant galaxy being magnified by the closer galaxy cluster," says Carlos De Breuck from ESO, a member of the team. De Breuck was making the observations at the APEX telescope on the plateau of Chajnantor at an altitude of 5000 m in the Chilean Andes. The new galaxy SMM J2135-0102 is so bright because of the massive galaxy cluster that lies in the foreground. The vast mass of this cluster bends the light of the more distant galaxy, acting as a gravitational lens [2]. As with a telescope, it magnifies and brightens our view of the distant galaxy. Thanks to a fortuitous alignment between the cluster and the distant galaxy, the latter is strongly magnified by a factor of 32. "The magnification

  9. The "Goldilocks Effect" in Cystic Fibrosis: identification of a lung phenotype in the cftr knockout and heterozygous mouse

    Directory of Open Access Journals (Sweden)

    Bates Jason HT

    2004-07-01

    Full Text Available Abstract Background Cystic Fibrosis is a pleiotropic disease in humans with primary morbidity and mortality associated with a lung disease phenotype. However, knockout in the mouse of cftr, the gene whose mutant alleles are responsible for cystic fibrosis, has previously failed to produce a readily, quantifiable lung phenotype. Results Using measurements of pulmonary mechanics, a definitive lung phenotype was demonstrated in the cftr-/- mouse. Lungs showed decreased compliance and increased airway resistance in young animals as compared to cftr+/+ littermates. These changes were noted in animals less than 60 days old, prior to any long term inflammatory effects that might occur, and are consistent with structural differences in the cftr-/- lungs. Surprisingly, the cftr+/- animals exhibited a lung phenotype distinct from either the homozygous normal or knockout genotypes. The heterozygous mice showed increased lung compliance and decreased airway resistance when compared to either homozygous phenotype, suggesting a heterozygous advantage that might explain the high frequency of this mutation in certain populations. Conclusions In the mouse the gene dosage of cftr results in distinct differences in pulmonary mechanics of the adult. Distinct phenotypes were demonstrated in each genotype, cftr-/-, cftr +/-, and cftr+/+. These results are consistent with a developmental role for CFTR in the lung.

  10. Novel apexification method in a non-vital tooth with an open apex: a case report.

    Directory of Open Access Journals (Sweden)

    Hamid Razavian

    2014-06-01

    Full Text Available Many materials have been introduced for apexification each having their own advantages and disadvantages. This case report aims to present a new method of apexification using a combination of deproteinized bovine bone mineral (DBBM and enamel matrix derivative (EMD. After irrigating the canal of the maxillary right canine with 2.5 % sodium hypochlorite, a mixture of Bio-Oss and EMD was packed into the apical region for formation of an apical barrier and the canal was obturated by thermoplastic gutta percha technique with AH26 sealer; coronal seal was achieved by resin bonded composite. The size of the periapical lesion decreased significantly after 3, 6, 12 and 18-months. The patient had no radiographic signs or clinical symptoms at 24-month follow up and complete maturation of the apex and healing of the periapical bone were achieved.

  11. Interdisciplinary Approach to a Tooth with Open Apex and Persistent Sinus

    Directory of Open Access Journals (Sweden)

    Anoop N. Das

    2015-01-01

    Full Text Available Traumatic injuries in childhood may disrupt root development leading to a tooth with open apex. Apexification procedures in such cases aim at root end closure after reasonable period of time. In some chronic cases, complete healing of the periapical area does not occur resulting in development of a nonhealing sinus. Failure of nonsurgical approach in such cases needs surgical intervention permitting thorough periapical curettage. In the present case, apexification procedure with MTA achieved root end closure but failed to heal the sinus for which surgical treatment was completed with thorough periapical curettage and application of platelet rich fibrin (PRF and a combination of β-tricalcium phosphate and hydroxyapatite resulted in healing.

  12. Search for a new gauge boson in the $A'$ Experiment (APEX)

    CERN Document Server

    Abrahamyan, S; Allada, K; Anez, D; Averett, T; Barbieri, A; Bartlett, K; Beacham, J; Bono, J; Boyce, J R; Brindza, P; Camsonne, A; Cranmer, K; Dalton, M M; deJager, C W; Donaghy, J; Essig, R; Field, C; Folts, E; Gasparian, A; Goeckner-Wald, N; Gomez, J; Graham, M; Hansen, J -O; Higinbotham, D W; Holmstrom, T; Huang, J; Iqbal, S; Jaros, J; Jensen, E; Kelleher, A; Khandaker, M; LeRose, J J; Lindgren, R; Liyanage, N; Long, E; Mammei, J; Markowitz, P; Maruyama, T; Maxwell, V; Mayilyan, S; McDonald, J; Michaels, R; Moffeit, K; Nelyubin, V; Odian, A; Oriunno, M; Partridge, R; Paolone, M; Piasetzky, E; Pomerantz, I; Qiang, Y; Riordan, S; Roblin, Y; Sawatzky, B; Schuster, P; Segal, J; Selvy, L; Shahinyan, A; Subedi, R; Sulkosky, V; Stepanyan, S; Toro, N; Walz, D; Wojtsekhowski, B; Zhang, J

    2011-01-01

    We present a search at Jefferson Laboratory for new forces mediated by sub-GeV vector bosons with weak coupling $\\alpha'$ to electrons. Such a particle $A'$ can be produced in electron-nucleus fixed-target scattering and then decay to an $e^+e^-$ pair, producing a narrow resonance in the QED trident spectrum. Using APEX test run data, we searched in the mass range 175--250 MeV, found no evidence for an $A'\\to e^+e^-$ reaction, and set an upper limit of $\\alpha'/\\alpha \\simeq 10^{-6}$. Our findings demonstrate that fixed-target searches can explore a new, wide, and important range of masses and couplings for sub-GeV forces.

  13. APEX: A Prime EXperiment at Jefferson Lab - Test Run Results and Full Run Plans; Update

    Energy Technology Data Exchange (ETDEWEB)

    Beacham, James [Ohio University, JLAB

    2015-06-01

    APEX is an experiment at Thomas Jefferson National Accelerator Facility (JLab) in Virginia, USA, that searches for a new gauge boson (A') with sub-GeV mass and coupling to ordinary matter of g' ~ (10-6 - 10⁻²)e. Electrons impinge upon a fixed target of high-Z material. An A' is produced via a process analogous to photon bremsstrahlung, decaying to an e⁺+e⁻ pair. A test run was held in July of 2010, covering mA' = 175 to 250 MeV and couplings g'/e > 10⁻³. A full run is approved and will cover mA' ~ 65 to 525 MeV and g'/e > 2.3 x 10⁻⁴, and is expected to occur sometime in 2016 or 2017.

  14. Impacts of dichroic prism coatings on radiometry of the airborne imaging spectrometer APEX.

    Science.gov (United States)

    Hueni, A; Schlaepfer, D; Jehle, M; Schaepman, M

    2014-08-20

    The generation of well-calibrated radiometric measurements from imaging spectrometer data requires careful consideration of all influencing factors, as well as an instrument calibration based on a detailed sensor model. Deviations of ambient parameters (i.e., pressure, humidity, temperature) from standard laboratory conditions during airborne operations can lead to biases that should be accounted for and properly compensated by using dedicated instrument models. This study introduces a model for the airborne imaging spectrometer airborne prism experiment (APEX), describing the impact of spectral shifts as well as polarization effects on the radiometric system response due to changing ambient parameters. Key issues are related to changing properties of the dichroic coating applied to the dispersing elements within the optical path. We present a model based on discrete numerical simulations. With the improved modeling approach, we predict radiometric biases with an root mean square error (RMSE) below 1%, leading to a substantial improvement of radiometric stability and predictability of system behavior. PMID:25321104

  15. Search for a new gauge boson in the $A'$ Experiment (APEX)

    Energy Technology Data Exchange (ETDEWEB)

    Abrahamyan, S; Allada, K; Anez, D; Averett, T; Barbieri, A; Bartlett, K; Beacham, J; Bono, J; Boyce, J R; Brindza, P; Camsonne, A; Cranmer, K; Dalton, M M; de Jager, C W; Donaghy, J; Essig, R; Field, C; Folts, E; Gasparian, A; Goeckner-Wald, N; Gomez, J; Graham, M; Hansen, J -O; Higinbotham, D W; Holmstrom, T; Huang, J; Iqbal, S; Jaros, J; Jensen, E; Kelleher, A; Khandaker, M; LeRose, J J; Lindgren, R; Liyanage, N; Long, E; Mammei, J; Markowitz, P; Maruyama, T; Maxwell, V; Mayilyan, S; McDonald, J; Michaels, R; Moffeit, K; Nelyubin, V; Odian, A; Oriunno, M; Partridge, R; Paolone, M; Piasetzky, E; Pomerantz, I; Qiang, Y; Riordan, S; Roblin, Y; Sawatzky, B; Schuster, P; Segal, J; Selvy, L; Shahinyan, A; Subedi, R; Sulkosky, V; Stepanyan, S; Toro, N; Walz, D; Wojtsekhowski, B

    2011-11-01

    We present a search at Jefferson Laboratory for new forces mediated by sub-GeV vector bosons with weak coupling {alpha}' to electrons. Such a particle A' can be produced in electron-nucleus fixed-target scattering and then decay to an e{sup +}e{sup -} pair, producing a narrow resonance in the QED trident spectrum. Using APEX test run data, we searched in the mass range 175-250 MeV, found no evidence for an A' {yields} e{sup +}e{sup -} reaction, and set an upper limit of {alpha}'/{alpha} {approx} 10{sup -6}. Our findings demonstrate that fixed-target searches can explore a new, wide, and important range of masses and couplings for sub-GeV forces.

  16. Bioremediation efficacy in Marrow Marsh following the Apex oil spill, Galveston Bay, Texas

    International Nuclear Information System (INIS)

    Samples taken from Marrow Marsh in Galveston Bay, Texas were taken to assess the efficacy of the August 5, 1990 bioremediation treatment in the marsh following the Apex barges oil spill on July 28, 1990. The bioremediation treatment combined a lyophilized bacterial mixture and a nutrient mix containing phosphorus and nitrogen. Samples from the marsh had been collected over a 96 h period from both treated and untreated oiled sites. Oil fingerprinting, fatty acid analysis, polynuclear aromatic hydrocarbons analysis, and total petroleum hydrocarbons analysis were performed to evaluate changes in the chemical characteristics of spilled oil. Results of analyses, although not statistically reliable, failed to support the occurrence of any definite chemical alteration in the spilled oil that could be attributed to the bioremediation treatment. The relatively short sampling period and the number of samples taken, however, may have been insufficient to document the efficacy of the overall bioremediation effect. 13 refs., 6 figs., 4 tabs

  17. Phloem long-distance delivery of FLOWERING LOCUS T (FT) to the apex.

    Science.gov (United States)

    Yoo, Soo-Cheul; Chen, Cheng; Rojas, Maria; Daimon, Yasufumi; Ham, Byung-Kook; Araki, Takashi; Lucas, William J

    2013-08-01

    Cucurbita moschata FLOWERING LOCUS T-LIKE 2 (hereafter FTL2) and Arabidopsis thaliana (Arabidopsis) FLOWERING LOCUS T (FT), components of the plant florigenic signaling system, move long-distance through the phloem from source leaves to the vegetative apex where they mediate floral induction. The mechanisms involved in long-distance trafficking of FT/FTL2 remain to be elucidated. In this study, we identified the critical motifs on both FT and FTL2 required for cell-to-cell trafficking through mutant analyses using a zucchini yellow mosaic virus expression vector. Western blot analysis, performed on phloem sap collected from just beneath the vegetative apex of C. moschata plants, established that all mutant proteins tested retained the ability to enter the phloem translocation stream. However, immunolocalization studies revealed that a number of these FTL2/FT mutants were defective in the post-phloem zone, suggesting that a regulation mechanism for FT trafficking exists in the post-phloem unloading step. The selective movements of FT/FTL2 were further observed by microinjection and trichome rescue studies, which revealed that FT/FTL2 has the ability to dilate plasmodesmata microchannels during the process of cell-to-cell trafficking, and various mutants were compromised in their capacity to traffic through plasmodesmata. Based on these findings, a model is presented to account for the mechanism by which FT/FTL2 enters the phloem translocation stream and subsequently exits the phloem and enters the apical tissue, where it initiates the vegetative to floral transition.

  18. Contrasting patterns of individual specialization and trophic coupling in two marine apex predators.

    Science.gov (United States)

    Matich, Philip; Heithaus, Michael R; Layman, Craig A

    2011-01-01

    1. Apex predators are often assumed to be dietary generalists and, by feeding on prey from multiple basal nutrient sources, serve to couple discrete food webs. But there is increasing evidence that individual level dietary specialization may be common in many species, and this has not been investigated for many marine apex predators. 2. Because of their position at or near the top of many marine food webs, and the possibility that they can affect populations of their prey and induce trophic cascades, it is important to understand patterns of dietary specialization in shark populations. 3. Stable isotope values from body tissues with different turnover rates were used to quantify patterns of individual specialization in two species of 'generalist' sharks (bull sharks, Carcharhinus leucas, and tiger sharks, Galeocerdo cuvier). 4. Despite wide population-level isotopic niche breadths in both species, isotopic values of individual tiger sharks varied across tissues with different turnover rates. The population niche breadth was explained mostly by variation within individuals suggesting tiger sharks are true generalists. In contrast, isotope values of individual bull sharks were stable through time and their wide population level niche breadth was explained by variation among specialist individuals. 5. Relative resource abundance and spatial variation in food-predation risk tradeoffs may explain the differences in patterns of specialization between shark species. 6. The differences in individual dietary specialization between tiger sharks and bull sharks results in different functional roles in coupling or compartmentalizing distinct food webs. 7. Individual specialization may be an important feature of trophic dynamics of highly mobile marine top predators and should be explicitly considered in studies of marine food webs and the ecological role of top predators. PMID:20831730

  19. Into the deep: the functionality of mesopelagic excursions by an oceanic apex predator.

    Science.gov (United States)

    Howey, Lucy A; Tolentino, Emily R; Papastamatiou, Yannis P; Brooks, Edward J; Abercrombie, Debra L; Watanabe, Yuuki Y; Williams, Sean; Brooks, Annabelle; Chapman, Demian D; Jordan, Lance K B

    2016-08-01

    Comprehension of ecological processes in marine animals requires information regarding dynamic vertical habitat use. While many pelagic predators primarily associate with epipelagic waters, some species routinely dive beyond the deep scattering layer. Actuation for exploiting these aphotic habitats remains largely unknown. Recent telemetry data from oceanic whitetip sharks (Carcharhinus longimanus) in the Atlantic show a strong association with warm waters (>20°C) less than 200 m. Yet, individuals regularly exhibit excursions into the meso- and bathypelagic zone. In order to examine deep-diving behavior in oceanic whitetip sharks, we physically recovered 16 pop-up satellite archival tags and analyzed the high-resolution depth and temperature data. Diving behavior was evaluated in the context of plausible functional behavior hypotheses including interactive behaviors, energy conservation, thermoregulation, navigation, and foraging. Mesopelagic excursions (n = 610) occurred throughout the entire migratory circuit in all individuals, with no indication of site specificity. Six depth-versus-time descent and ascent profiles were identified. Descent profile shapes showed little association with examined environmental variables. Contrastingly, ascent profile shapes were related to environmental factors and appear to represent unique behavioral responses to abiotic conditions present at the dive apex. However, environmental conditions may not be the sole factors influencing ascents, as ascent mode may be linked to intentional behaviors. While dive functionality remains unconfirmed, our study suggests that mesopelagic excursions relate to active foraging behavior or navigation. Dive timing, prey constituents, and dive shape support foraging as the most viable hypothesis for mesopelagic excursions, indicating that the oceanic whitetip shark may regularly survey extreme environments (deep depths, low temperatures) as a foraging strategy. At the apex of these deep

  20. Into the deep: the functionality of mesopelagic excursions by an oceanic apex predator.

    Science.gov (United States)

    Howey, Lucy A; Tolentino, Emily R; Papastamatiou, Yannis P; Brooks, Edward J; Abercrombie, Debra L; Watanabe, Yuuki Y; Williams, Sean; Brooks, Annabelle; Chapman, Demian D; Jordan, Lance K B

    2016-08-01

    Comprehension of ecological processes in marine animals requires information regarding dynamic vertical habitat use. While many pelagic predators primarily associate with epipelagic waters, some species routinely dive beyond the deep scattering layer. Actuation for exploiting these aphotic habitats remains largely unknown. Recent telemetry data from oceanic whitetip sharks (Carcharhinus longimanus) in the Atlantic show a strong association with warm waters (>20°C) less than 200 m. Yet, individuals regularly exhibit excursions into the meso- and bathypelagic zone. In order to examine deep-diving behavior in oceanic whitetip sharks, we physically recovered 16 pop-up satellite archival tags and analyzed the high-resolution depth and temperature data. Diving behavior was evaluated in the context of plausible functional behavior hypotheses including interactive behaviors, energy conservation, thermoregulation, navigation, and foraging. Mesopelagic excursions (n = 610) occurred throughout the entire migratory circuit in all individuals, with no indication of site specificity. Six depth-versus-time descent and ascent profiles were identified. Descent profile shapes showed little association with examined environmental variables. Contrastingly, ascent profile shapes were related to environmental factors and appear to represent unique behavioral responses to abiotic conditions present at the dive apex. However, environmental conditions may not be the sole factors influencing ascents, as ascent mode may be linked to intentional behaviors. While dive functionality remains unconfirmed, our study suggests that mesopelagic excursions relate to active foraging behavior or navigation. Dive timing, prey constituents, and dive shape support foraging as the most viable hypothesis for mesopelagic excursions, indicating that the oceanic whitetip shark may regularly survey extreme environments (deep depths, low temperatures) as a foraging strategy. At the apex of these deep

  1. Comparative evaluation of the accuracy of two electronic apex locators in determining the working length in teeth with simulated apical root resorption: An in vitro study

    Science.gov (United States)

    Saraswathi, Vidya; Kedia, Archit; Purayil, Tina Puthen; Ballal, Vasudev; Saini, Aakriti

    2016-01-01

    Introduction: Accurate determination of working length (WL) is a critical factor for endodontic success. This is commonly achieved using an apex locator which is influenced by the presence or absence of the apical constriction. Hence, this study was done to compare the accuracy of two generations of apex locators in teeth with simulated apical root resorption. Materials and Methods: Forty maxillary central incisors were selected and after access preparation, were embedded in an alginate mold. On achieving partial set, teeth were removed, and a 45° oblique cut was made at the apex. The teeth were replanted and stabilized in the mold, and WL was determined using two generations of apex locators (Raypex 5 and Apex NRG XFR). Actual length of teeth (control) was determined by visual method. Statistical Analysis: Results were subjected to statistical analysis using the paired t-test. Results: Raypex 5 and Apex NRG was accurate for only 33.75% and 23.75% of samples, respectively. However, with ±0.5 mm acceptance limit, they showed an average accuracy of 56.2% and 57.5%, respectively. There was no significant difference in the accuracy between the two apex locators. Conclusion: Neither of the two apex locators were 100% accurate in determining the WL. PMID:27656055

  2. The DNA Repair Enzyme Apurinic/Apyrimidinic Endonuclease (Apex Nuclease 2 Has the Potential to Protect against Down-Regulation of Chondrocyte Activity in Osteoarthritis

    Directory of Open Access Journals (Sweden)

    Naoko Yui

    2014-08-01

    Full Text Available Apurinic/apyrimidinic endonuclease 2 (Apex 2 plays a critical role in DNA repair caused by oxidative damage in a variety of human somatic cells. We speculated that chondrocyte Apex 2 may protect against the catabolic process of articular cartilage in osteoarthritis (OA. Higher levels of Apex 2 expression were histologically observed in severely compared with mildly degenerated OA cartilage from STR/OrtCrlj mice, an experimental model which spontaneously develops OA. The immunopositivity of Apex 2 was significantly correlated with the degree of cartilage degeneration. Moreover, the OA-related catabolic factor interleukin-1β induced the expression of Apex 2 in chondrocytes, while Apex 2 silencing using small interfering RNA reduced chondrocyte activity in vitro. The expression of Apex 2 in chondrocytes therefore appears to be associated with the degeneration of articular cartilage and could be induced by an OA-related catabolic factor to protect against the catabolic process of articular cartilage. Our findings suggest that Apex 2 may have the potential to prevent the catabolic stress-mediated down-regulation of chondrocyte activity in OA.

  3. APEX simulation of runoff and total phosphorous for three adjacent row-crop watersheds with claypan soils

    Science.gov (United States)

    The Agricultural Policy Environmental Extender (APEX) model can simulate crop yields, runoff, and the transport of sediment and nutrients in small watersheds that have combinations of farm level landscapes, cropping systems and/or management practices. The objectives of the study were to parameteri...

  4. APEX sensitivity to atrazine dissipation rate on surface runoff loss within a coastal zone in Southeastern Puerto Rico

    Science.gov (United States)

    Simulation models are increasingly used to predict effects of conservation practices on transport of pesticides to water bodies. We used two models - the Agricultural Policy/Environmental eXtender (APEX) and the Riparian Ecosystem Management Model (REMM) to predict the movement of the herbicide, at...

  5. 76 FR 67243 - In the Matter of Accesspoint Corp., Aero Performance Products, Inc., Apex Resources Group, Inc...

    Science.gov (United States)

    2011-10-31

    ... COMMISSION In the Matter of Accesspoint Corp., Aero Performance Products, Inc., Apex Resources Group, Inc., Aradyme Corp., Bancroft Uranium, Inc., Fightersoft Multimedia Corp., Fortress Financial Group, Inc., and... Resources Group, Inc. because it has not filed any periodic reports since the period ended March 31,...

  6. Comparison of digital radiography and apex locator with the conventional method in root length determination of primary teeth

    Directory of Open Access Journals (Sweden)

    I E Neena

    2011-01-01

    Full Text Available Aim: The purpose of this study was to compare the Working length in primary teeth endodontics using intra oral digital radiovisiography and apex locator with conventional method for accuracy. Materials and Methods: This in vivo study was conducted on 30 primary teeth which were indicated for pulpectomy in the patients of the age group of 5-11 years All experimental teeth had adequate remaining tooth structure for rubber dam isolation and radiographicaly visible canals. Endodontic treatment was required due to irreversible pulpitis or pulp necrosis. A standardized intraoral periapical radiograph of the tooth was taken using conventional method by paralleling technique. The distance between the source and the tooth, tooth and the films were standardized using X-ray positioning device. During the pulpectomy procedure, the working length was determined by digital radiograph and apex locator. The measurements were then compared with the conventional method of root canal measurement technique for accuracy Result: From the results obtained we can conclude that Working length determined in primary molars using digital radiography and Apex locator did not show any significant difference in the mean working length measurements when compared with the conventional radiographic method. Conclusions: Apex locator is comparable to conventional radiograph in determining the working length without radiation in the primary teeth. Intraoral digital radiography is the safest method in determining the working length with significant reduction in radiation exposure.Hence, both the techniques can be safely used as alternatives to conventional radiographic methods in determining working length in primary teeth.

  7. Clinical Comparison of Root Length Measurements with Electronic Apex Locator and Conventional Radiography in Mandibular Deciduous Teeth

    Directory of Open Access Journals (Sweden)

    Eskandarian T.

    2011-04-01

    Full Text Available Statement of Problem: Success in pulpectomy of deciduous teeth greatly depends on the accuracy of root length measurements which have mostly been done radiographically. However, X-ray risks and patient cooperation have usually been a challenge for the clinicians.Purpose: The clinical comparison of root length measurements with electronic apex locator and conventional radiography in 4-6 year old children's mandibular deciduous teeth was the aim of the present study.Materials and Method: In the current clinical trial, 15 mandibular molars with 60 canals in 4-6 year old patients who had the treatment plan of pulpectomy were chosen. The measured root lengths with apex locator and parallel technique radiography were evaluated with the same reference point. Data were analyzed using simple linear regression, coefficient of correlation, coefficient of variability and also graphic Bland Altman Plot.Results: The accuracy of electronic apex locator measurements in ±0/5 from apical foramen was 85%. In all cases without considering pulp situation, the difference between the two techniques was not significant.Conclusion: The electronic apex locators are recommended for root length measurements of the deciduous mandibular molars without apical resorption, disregarding the pulp status, especially when initial radiographic films are available.

  8. Piroxicam treatment augments bone abnormalities in interleukin-10 knockout mice

    DEFF Research Database (Denmark)

    Holgersen, Kristine; Dobie, Ross; Farquharson, Colin;

    2015-01-01

    inflammation in an experimental colitis model. METHODS: Colitis was induced in interleukin-10 knockout mice (PAC IL-10 k.o.) by peroral administration of piroxicam for 12 days. The degree of colitis was assessed by clinical, macroscopic, and microscopic evaluation. Trabecular and cortical bone...... microarchitecture of tibia were determined using micro-computed tomography. Moreover, the serum levels of bone formation and bone resorption biomarkers were measured, and inflammatory protein profiling was performed on colons. RESULTS: PAC IL-10 k.o. mice developed severe colitis, characterized by hyperplasia...... and focal transmural inflammation, which was consistent with Crohn's disease-like pathology. The gut inflammation was accompanied by a 14% and 12% reduction in trabecular thickness relative to piroxicam-treated wild type and untreated wild type mice, respectively (P

  9. Antiatherogenic effects of oleanolic acid in apolipoprotein E knockout mice

    DEFF Research Database (Denmark)

    Buus, Niels Henrik; Hansson, Nicolaj Christopher; Rodriguez-Rodriguez, Rosalia;

    2011-01-01

    Oleanolic acid (OA) is a plant triterpenoid steroid with potentially antiatherogenic properties. We investigated whether OA affected atherosclerosis development and vascular function in apolipoprotein E knockout (ApoE(-/-)) mice. ApoE(-/-) mice were fed a high cholesterol Western-type diet...... in combination with OA (100 mg/kg/day), fluvastatin (5 mg/kg/day) or vehicle, with wild type (WT) mice serving as controls. After 8 weeks of treatment atherosclerotic plaque areas in the aortic arch and plasma lipid concentrations were determined. Vasoconstriction and relaxation of the proximal part of aorta...... were investigated in vitro. Inducible nitric oxide synthase (iNOS) was visualized using immunoblotting. As opposed to WT and fluvastatin- and vehicle-treated mice, OA-fed ApoE(-/-) mice gained no weight during the treatment period. Plasma concentrations of total-cholesterol and triglyceride were...

  10. Medium effects on spin observables of proton knockout reactions

    Energy Technology Data Exchange (ETDEWEB)

    Krein, G. [Instituto de Fisica Teorica (IFT), Sao Paulo, SP (Brazil); Maris, T.A.J.; Rodrigues, B.B.; Veit, E.A. [Rio Grande do Sul Univ., Porto Alegre, RS (Brazil). Inst. de Fisica

    1994-07-01

    Medium modifications of the properties of bound nucleons and mesons are investigated by means of medium energy quasi free proton knockout reactions with polarized incident protons. The sensitivity of the spin observables of these reactions to modifications of the nucleon and meson properties is studied using the Bonn one-boson exchange model of the nucleon-nucleon interaction. A method proposed to extract the pp analysing power in medium from the (p, 2 p) asymmetries indicates a reduction of this quantity compared to its free space value. This reduction is linked to modifications of masses and coupling constants of the nucleons and mesons in the nucleus. The implications of these modifications for another spin observable to be measured in the future are discussed. (author). 39 refs, 9 figs.

  11. RF-knockout Extraction System for the CNAO Synchrotron

    CERN Document Server

    Carmignani, Nicola; Serio, Mario; Balbinot, Giovanni; Bressi, Erminia; Caldara, Michele; Pullia, Marco; Bosser, Jacques; Venchi, Giuseppe

    2010-01-01

    The National Centre for Oncological Hadrontherapy (CNAO) is a centre in Italy for the treatment of patients affected by tumours with proton and carbon ions beams accelerated in a synchrotron. The synchrotron extraction method is based on the use of a betatron core. This work aims to verify, through a theoretical study and a simulation, the possibility of using the RF-knockout extraction method exploiting the existing hardware. A simulation program has been written to simulate the extraction system of the synchrotron with the purpose to define the parameters of the radio frequency. Two types of radio frequencies have been compared in order to obtain a constant spill with the minimum ripple: a carrier wave with a frequency and amplitude modulation, and a gaussian narrow band noise modulated in amplitude. Results of the simulation and considerations on the kicker characteristics are presented

  12. SNARE function analyzed in synaptobrevin/VAMP knockout mice.

    Science.gov (United States)

    Schoch, S; Deák, F; Königstorfer, A; Mozhayeva, M; Sara, Y; Südhof, T C; Kavalali, E T

    2001-11-01

    SNAREs (soluble NSF-attachment protein receptors) are generally acknowledged as central components of membrane fusion reactions, but their precise function has remained enigmatic. Competing hypotheses suggest roles for SNAREs in mediating the specificity of fusion, catalyzing fusion, or actually executing fusion. We generated knockout mice lacking synaptobrevin/VAMP 2, the vesicular SNARE protein responsible for synaptic vesicle fusion in forebrain synapses, to make use of the exquisite temporal resolution of electrophysiology in measuring fusion. In the absence of synaptobrevin 2, spontaneous synaptic vesicle fusion and fusion induced by hypertonic sucrose were decreased approximately 10-fold, but fast Ca2+-triggered fusion was decreased more than 100-fold. Thus, synaptobrevin 2 may function in catalyzing fusion reactions and stabilizing fusion intermediates but is not absolutely required for synaptic fusion.

  13. Delayed liver regeneration after partial hepatectomy in adiponectin knockout mice

    International Nuclear Information System (INIS)

    We previously demonstrated that adiponectin has anti-fibrogenic and anti-inflammatory effects in the liver of mouse models of various liver diseases. However, its role in liver regeneration remains unclear. The aim of this study was to determine the role of adiponectin in liver regeneration. We assessed liver regeneration after partial hepatectomy in wild-type (WT) and adiponectin knockout (KO) mice. We analyzed DNA replication and various signaling pathways involved in cell proliferation and metabolism. Adiponectin KO mice exhibited delayed DNA replication and increased lipid accumulation in the regenerating liver. The expression levels of peroxisome proliferator-activated receptor (PPAR) α and carnitine palmitoyltransferase-1 (CPT-1), a key enzyme in mitochondrial fatty acid oxidation, were decreased in adiponectin KO mice, suggesting possible contribution of altered fat metabolism to these phenomena. Collectively, the present results highlight a new role for adiponectin in the process of liver regeneration.

  14. Reduced Extinction of Hippocampal-Dependent Memories in CPEB Knockout Mice

    Science.gov (United States)

    Zearfoss, N. Ruth; Richter, Joel D.; Berger-Sweeney, Joanne

    2006-01-01

    CPEB is a sequence-specific RNA binding protein that regulates translation at synapses. In neurons of CPEB knockout mice, synaptic efficacy is reduced. Here, we have performed a battery of behavioral tests and find that relative to wild-type animals, CPEB knockout mice, although similar on many baseline behaviors, have reduced extinction of…

  15. 植物乳杆菌对IL-10基因敲除结肠炎小鼠黏附分子的影响%Lactobacillus plantarum attenuates up-regulation of adhesion molecules expression in IL-10 gene knockout mice with colitis

    Institute of Scientific and Technical Information of China (English)

    储昭新; 陈红旗; 张明; 秦环龙

    2009-01-01

    目的: 观察植物乳杆菌( Lactobacillus plantarum,LP)灌胃对IL-10基因敲除小鼠肠道炎症和淋巴细胞归巢的影响.方法: 取IL-10基因敲除(knockout,KO)小鼠和未作基因敲除的背景鼠分为4组: 对照组(野生组WT)、加植物乳杆菌组(WT+LP)、IL-10基因敲除模型组(KO)、模型加植物乳杆菌组(KO+LP).4 wk开始对照组和KO组每日予PBS灌胃,WT+LP和KO+LP组予溶于PBS的LP灌胃,持续4-8 wk结束.实验结束后取各组小鼠结肠行炎症评分和电镜亚显微结构观察,并用RT-PCR和Western blot检测归巢相关分子MAdCAM-1、ICAM-1、α4β7及CD3的表达.结果: 8 w k 后K O小鼠1 0 0%发生肠道炎症,且其CD3及黏附分子α4β7、ICAM-1和MAdCAM-1的mRNA和蛋白表达水平较WT组均明显增高(mRNA: t = 39.42,8.83,25.53,45.78,均P<0.01;CD3、ICAM-1、MAdCAM-1蛋白: t = 19.04,29.57,12.29,均P<0.01).予以益生菌LP灌胃后,KO+LP组小鼠CD3及黏附分子α4β7、ICAM-1和MAdCAM-1的mRNA和蛋白表达水平较KO组均明显降低(mRNA: t = 20.34;4.95;14.21;22.31,均P<0.01;CD3、ICAM-1、MAdCAM-1蛋白: t = 6.82,14.10,7.03,均P<0.01);WT+L P组小鼠CD3及黏附分子α4β7、ICAM-1和MAdCAM-1的mRNA较WT组均明显降低( t = 9.33,10.55,7.75,6.69,均P<0.01),而WT+LP组小鼠CD3及黏附分子ICAM-1和MAdCAM-1的蛋白表达水平无明显降低.结论: 植物乳杆菌能下调黏附分子在IL-10基因敲除结肠炎小鼠中的高表达,这可能是其减轻炎症状态,缓解炎症性肠病的重要机制之一.

  16. Preaxial Polydactyly in Sost/Sostdc1 Double Knockouts

    Energy Technology Data Exchange (ETDEWEB)

    Yee, C M; Collette, N M; Loots, G G

    2011-07-29

    In the United States, {approx}5% are born with congenital birth defects due to abnormal function of cellular processes and interactions. Sclerosteosis, a rare autosomal recessive disease, causes hyperostosis of the axial and appendicular skeleton, and patients present radial deviation, digit syndactyly, nail dysplasia, and overall high bone mineral density. Sclerosteosis is due to a loss of function of sclerostin (Sost). Sost is a Wnt (abbrev.) antagonist; when mutated, nonfunctional Sost results in hyperactive osteoblast activity which leads to abnormal high bone mass. Previous studies have shown that Sost overexpression in transgenic mice causes reduced bone mineral density and a variety of limb phenotypes ranging from lost, fused, and split phalanges. Consistent with clinical manifestations of Sclerosteosis, Sost knockout mice exhibit increased generalized bone mineral density and syndactyly of the digits. Sostdc1 is a paralog of Sost that has also been described as an antagonist of Wnt signaling, in developing tooth buds. Unlike Sost knockouts, Sostdc1 null mice do not display any limb abnormalities. To determine if Sost and Sostdc1 have redundant functions during limb patterning, we examined Sost; Sostdc1 mice determined that they exhibit a novel preaxial polydactyly phenotype with a low penetrance. LacZ staining, skeletal preparations, and in situ hybridization experiments were used to help characterize this novel phenotype and understand how this phenotype develops. We find Sost and Sostdc1 to have complementary expression patterns during limb development, and the loss of their expression alters the transcription of several key limb regulators, such as Fgf8, Shh and Grem.

  17. Differences in Interactions between Azole Drugs Related to Modifications in the 14-α Sterol Demethylase Gene (cyp51A) of Aspergillus fumigatus

    OpenAIRE

    Garcia-Effron, G.; Mellado, E.; A. Gomez-Lopez; Alcazar-Fuoli, L.; Cuenca-Estrella, M.; Rodriguez-Tudela, Juan L.

    2005-01-01

    The combined activity of different azole drugs was investigated. Thirty-one Aspergillus fumigatus strains were tested, including two cyp51A− and one cyp51B− gene-knockout strain and azole-susceptible and -resistant strains with different resistance mechanisms. The combination of itraconazole and voriconazole was synergistic for all strains except for those with gene knockouts.

  18. Apolipoprotein E gene knock-out and high-fat diet on IP3 and IP3R-1 expression in neurons of mice hippocampal CA1 and CA3%载脂蛋白E基因敲除及高脂饮食小鼠海马CA1和CA3区神经元内IP3及IP3R-1表达的变化

    Institute of Scientific and Technical Information of China (English)

    周祎; 刘娟; 黄大可; 桂丽; 汪渊; 贾雪梅

    2011-01-01

    Objective To observe the change of IP3 and IP3R-1's expression in neurons of mice's hippocampal CA1 and CA3 , which had been treated by Apolipoprotein E gene knock-out( ApoE KO ) and high-fat diet. Methods 30 C57BL/6J mice were divided int0 3 groups: the control group ( C group ), ApoE KO group ( KO group ),ApoE KO high-fat diet group ( KO-HF group ). After mice model established, weight and plasma lipid of these mice were measured. The brain tissues of the mice were observed by HE staining, immunohistochemistry staining,and computer image analysis. Results The weight, total cholesterol, triglyceride . low-density lipoprotein cholesterol of the KO and KO-HF groups were all higher than those in the control group( P < 0. 05 ). The H-E staining showed that, in the KO and KO-HF groups, the pyramidal cell layers ranged sparse and cell body were relatively small. Compared with C group, The average optical density of IP3 and IP3R-1 in neurons of hippocampal CA1 and CA3 in the KO group reduced. The average optical density in the KO-HF group reduced obviously( P < 0. 05 ).Conclusion ApoE KO and high-fat diet can decrease the expression level of IP3 and IP3 R-1 in neurons of hippocampal CA1 and CA3. These two proteins ( IP3 and IP3R-1 ) might take participate in the pathologic process in Alzheimer disease which caused by the abnormal ApoE.%目的 观察载脂蛋白E(ApoE)基因敲除(KO)及高脂饮食小鼠海马CA1和CA3区神经元内三磷酸肌醇(IP3)和三磷酸肌醇受体-Ⅰ(IP3R-1)表达的变化.方法 将30只C57BL/6J小鼠分为对照组(C组)、ApoE KO组(KO组)、ApoE KO高脂饮食组(KO-HF组).小鼠造模成功后称重;取血检测血脂;取小鼠脑组织分别进行HE染色、免疫组织化学染色和计算机图像分析.结果 与C组比较,KO、KO-HF组体重、总胆固醇、甘油三酯及低密度脂蛋白胆固醇含量明显升高(P<0.05).HE染色观察到,KO和KO-HF组小鼠海马锥体细胞排列较

  19. Conditional (intestinal-specific) knockout of the riboflavin transporter-3 (RFVT-3) impairs riboflavin absorption.

    Science.gov (United States)

    Subramanian, Veedamali S; Lambrecht, Nils; Lytle, Christian; Said, Hamid M

    2016-02-15

    Riboflavin (RF) is indispensable for normal cell metabolism, proliferation, and growth. The RFVT-3 protein (product of the Slc52a3 gene) is expressed in the gut with the expression being restricted to the apical membrane domain of the polarized intestinal epithelial cells. The relative contribution of RFVT-3 to total carrier-mediated RF uptake in the native intestine, however, is not clear. We addressed this issue in the current investigation using a conditional (intestinal-specific) RFVT-3 knockout (cKO) mouse model developed by the Cre/Lox approach. All RFVT-3 cKO mice were found to be RF deficient and showed a significant growth and development retardation; also, nearly two-thirds of them died prematurely between the age of 6 and 12 wk. In vivo (intestinal and colonic loops) and in vitro (native isolated intestinal epithelial cells) uptake studies showed a severe inhibition in carrier-mediated RF uptake in the cKO mice compared with control littermates. We also observed a significant increase in the level of expression of oxidative stress-responsive genes in the intestine of the cKO mice compared with control littermates. Supplementation of the RFVT-3 cKO mice with pharmacological doses of RF led to a complete correction of the growth retardation and to normalization in the level of expression of the oxidative stress-responsive genes in the gut. These results show, for the first time, that the RFVT-3 system is the main transporter involved in carrier-mediated RF uptake in the native mouse small and large intestine, and that its dysfunction impairs normal RF body homeostasis.

  20. Highly efficient generation of GGTA1 biallelic knockout inbred mini-pigs with TALENs.

    Directory of Open Access Journals (Sweden)

    Jige Xin

    Full Text Available Inbred mini-pigs are ideal organ donors for future human xenotransplantations because of their clear genetic background, high homozygosity, and high inbreeding endurance. In this study, we chose fibroblast cells from a highly inbred pig line called Banna mini-pig inbred line (BMI as donor nuclei for nuclear transfer, combining with transcription activator-like effector nucleases (TALENs and successfully generated α-1,3-galactosyltransferase (GGTA1 gene biallelic knockout (KO pigs. To validate the efficiency of TALEN vectors, in vitro-transcribed TALEN mRNAs were microinjected into one-cell stage parthenogenetically activated porcine embryos. The efficiency of indel mutations at the GGTA1-targeting loci was as high as 73.1% (19/26 among the parthenogenetic blastocysts. TALENs were co-transfected into porcine fetal fibroblasts of BMI with a plasmid containing neomycin gene. The targeting efficiency reached 89.5% (187/209 among the survived cell clones after a 10 d selection. More remarkably 27.8% (58/209 of colonies were biallelic KO. Five fibroblast cell lines with biallelic KO were chosen as nuclear donors for somatic cell nuclear transfer (SCNT. Three miniature piglets with biallelic mutations of the GGTA1 gene were achieved. Gal epitopes on the surface of cells from all the three biallelic KO piglets were completely absent. The fibroblasts from the GGTA1 null piglets were more resistant to lysis by pooled complement-preserved normal human serum than those from wild-type pigs. These results indicate that a combination of TALENs technology with SCNT can generate biallelic KO pigs directly with high efficiency. The GGTA1 null piglets with inbred features created in this study can provide a new organ source for xenotransplantation research.

  1. Newly generated heparanase knock-out mice unravel co-regulation of heparanase and matrix metalloproteinases.

    Directory of Open Access Journals (Sweden)

    Eyal Zcharia

    Full Text Available BACKGROUND: Heparanase, a mammalian endo-beta-D-glucuronidase, specifically degrades heparan sulfate proteoglycans ubiquitously associated with the cell surface and extracellular matrix. This single gene encoded enzyme is over-expressed in most human cancers, promoting tumor metastasis and angiogenesis. PRINCIPAL FINDINGS: We report that targeted disruption of the murine heparanase gene eliminated heparanase enzymatic activity, resulting in accumulation of long heparan sulfate chains. Unexpectedly, the heparanase knockout (Hpse-KO mice were fertile, exhibited a normal life span and did not show prominent pathological alterations. The lack of major abnormalities is attributed to a marked elevation in the expression of matrix metalloproteinases, for example, MMP2 and MMP14 in the Hpse-KO liver and kidney. Co-regulation of heparanase and MMPs was also noted by a marked decrease in MMP (primarily MMP-2,-9 and 14 expression following transfection and over-expression of the heparanase gene in cultured human mammary carcinoma (MDA-MB-231 cells. Immunostaining (kidney tissue and chromatin immunoprecipitation (ChIP analysis (Hpse-KO mouse embryonic fibroblasts suggest that the newly discovered co-regulation of heparanase and MMPs is mediated by stabilization and transcriptional activity of beta-catenin. CONCLUSIONS/SIGNIFICANCE: The lack of heparanase expression and activity was accompanied by alterations in the expression level of MMP family members, primarily MMP-2 and MMP-14. It is conceivable that MMP-2 and MMP-14, which exert some of the effects elicited by heparanase (i.e., over branching of mammary glands, enhanced angiogenic response can compensate for its absence, in spite of their different enzymatic substrate. Generation of viable Hpse-KO mice lacking significant abnormalities may provide a promising indication for the use of heparanase as a target for drug development.

  2. Characterization of liquid entrainment in the AP1000 automatic depressurization system from APEX tests

    International Nuclear Information System (INIS)

    Full text of publication follows: The AP1000 is a 1000 MWe advanced nuclear power plant that uses passive safety features to enhance plant safety and to provide significant and measurable improvements in plant simplification, reliability, investment protection and plant costs. The AP1000 relies heavily on the 600 MWe AP600 which received design certification in 1999. A critical part of the AP600 design certification process involved the testing of the passive safety systems. A one-fourth height, one-fourth pressure test facility, APEX-600, was constructed at the Oregon State University to study design basis events, and to provide a body of data to be used to validate the computer models used to analyze the AP600. This facility was extensively modified to reflect the design changes for AP1000 including higher power in the electrically heated rods representing the reactor core, and changes in the size of the pressurizer, core makeup tanks and automatic depressurization system. The APEX-1000 test facility was used to perform design basis accident simulations and separate effects tests to support the AP1000 design certification process. In the event of a LOCA, the AP1000 passive core cooling system provides sources of core makeup water along with an automatic depressurization system (ADS) consisting of several stages of valves which reduce the reactor coolant system pressure in a controlled manner. The final stage of this system, ADS-4, consists of four large valves that open off the hot legs, reducing the pressure to allow gravity injection from the in-containment refueling water storage tank (IRWST) and eventually the containment sump. The 67% increase in power from AP600 to AP1000 results in proportionally larger steam velocities exiting the core. Higher steam velocities could increases the potential for significant liquid entrainment out the ADS-4 lines, affecting the liquid inventory in the reactor. Tests were performed in APEX-1000 to characterize the two

  3. Characterization of liquid entrainment in the AP1000 automatic depressurization system from APEX tests

    Energy Technology Data Exchange (ETDEWEB)

    Richard F Wright; Terry L Schulz [Westinghouse Electric Co., PO Box 355, Pittsburgh, PA (United States); Jose N Reyes; John Groome [Oregon State University, Corvallis, OR (United States)

    2005-07-01

    Full text of publication follows: The AP1000 is a 1000 MWe advanced nuclear power plant that uses passive safety features to enhance plant safety and to provide significant and measurable improvements in plant simplification, reliability, investment protection and plant costs. The AP1000 relies heavily on the 600 MWe AP600 which received design certification in 1999. A critical part of the AP600 design certification process involved the testing of the passive safety systems. A one-fourth height, one-fourth pressure test facility, APEX-600, was constructed at the Oregon State University to study design basis events, and to provide a body of data to be used to validate the computer models used to analyze the AP600. This facility was extensively modified to reflect the design changes for AP1000 including higher power in the electrically heated rods representing the reactor core, and changes in the size of the pressurizer, core makeup tanks and automatic depressurization system. The APEX-1000 test facility was used to perform design basis accident simulations and separate effects tests to support the AP1000 design certification process. In the event of a LOCA, the AP1000 passive core cooling system provides sources of core makeup water along with an automatic depressurization system (ADS) consisting of several stages of valves which reduce the reactor coolant system pressure in a controlled manner. The final stage of this system, ADS-4, consists of four large valves that open off the hot legs, reducing the pressure to allow gravity injection from the in-containment refueling water storage tank (IRWST) and eventually the containment sump. The 67% increase in power from AP600 to AP1000 results in proportionally larger steam velocities exiting the core. Higher steam velocities could increases the potential for significant liquid entrainment out the ADS-4 lines, affecting the liquid inventory in the reactor. Tests were performed in APEX-1000 to characterize the two

  4. 超声生物显微镜斑点追踪成像评价EP3基因敲除小鼠心脏的功能%Assessment of heart functions of EP3 gene knockout mouse with ultrasound biomicroscopy and speckle tracking imaging

    Institute of Scientific and Technical Information of China (English)

    季亚玮; 盛红专; 桂乐; 黄竺筠; 吴翔; 朱健华

    2016-01-01

    Objective: To investigate the wild type(WT) knockout EP3(EP3-/-) mouse cardiac structure and function, ultrasound biomicroscopy(UBM) and speckle tracking echocardiography(STE) were applied, and to understand the important role of EP3 receptor in the changes of dilated cardiomyopathy disease course. Methods: Take conventional ultrasound echocardiography inspection of twenty mice of two groups using the VisualSonics Vevo 2100 Imaging System, collecting two-dimensional(2D) images such as long axis of parasternal left ventricular,short axis of left ventricular papillary muscle level, short axis of aorta and the M mode ultrasound of above 2D images. Conventional ultrasound echocardiography included heart rate(HR), left ventricular end systolic diameter(LVIDs), left ventricular diastolic diameter(LVIDd), left ventricular end systolic anterior wall thickness(LVAWs), left ventricular end diastolic anterior wall thickness(LVAWd), left ventricular posterior wall thickness(LVPWs), left ventricular diastolic posterior wall thickness(LVPWd), left ventricular volume(LV Vols), left ventricular ejection fraction(LVEF), fractional shortening(FS), left ventricular mass(LV mass), mitral E/A(MV E/A), isovolumic relaxation time(IVRT). To evaluate the endocardial and epicardial left ventricular strain by using ultrasound biomicroscopy speckle tracking imaging system and analysis the peak strain of two different sections of left ventricular, epicardial longitudinal strain, circumferential strain and radial strain time to peak(TPK), maximum peak delay time of ventricular wall(MOWD). Results: After examination of 2D and M mode ultrasound, heart function of EP3-/- mice was significantly lower than that of WT mice. Analysis of endocardial and epicardial of left ventricular myocardial strain showed the decreasing trend of the longitudinal, circumferential and radial strain in EP3-/- mice than WT mice. Conclusions: UBM can detect the mouse dilated cardiomyopathy and the structure change of

  5. BRN 3.1 Knockouts Affect the Vestibular, Autonomic, and Circadian Rhythm Responses to 2G Exposure

    Science.gov (United States)

    Murakami, D. M.; Erkman, L.; Rosenfeld, M. G.; Fuller, C. A.

    1999-01-01

    Our previous studies have demonstrated that 2G exposure via centrifugation significantly attenuated the daily mean and circadian rhythm amplitude of rat body temperature (Tb), heart rate, and activity (Act). In addition, 2G exposure activates neural responses in several vestibular, autonomic, and circadian nuclei. Although we have characterized the effect of 2G on an animal's physiological, neuronal, and behavioral responses, it will be important to understand the underlying neural and physiological mechanisms that mediate those responses. For example, the vestibular responses, proprioceptive feedback, or fluid shifts may be the critical factors that mediate the responses to 2G. As a first step to understand the relative importance of these different response pathways to altered gravitational fields, this study examined the contribution of the vestibular system by utilizing an animal model from molecular biology. Brain 3.1 (Bm 3.1) is a POU domain homeobox gene involved in the normal development of the vestibular and auditory system. Brn 3.1 deletion results in a loss of hair cells in the otoliths, semicircular canals, and cochlea. As a result mice with a Brn 3.1 deletion do not have a functioning vestibular or auditory system. The BRN 3.1 knockout mouse could be a very useful animal model for isolating the role of the vestibular system in mediating the physiological responses to 2G exposure. Therefore, this study compared the effect of 2G exposure via centrifugation between Brn 3.1 knockout (KO) versus Wildtype (W) mice.

  6. Effects of Chronic Mild Stress on the Development of Atherosclerosis and Expression of Toll-Like Receptor 4 Signaling Pathway in Adolescent Apolipoprotein E Knockout Mice

    Directory of Open Access Journals (Sweden)

    Hongfeng Gu

    2009-01-01

    Full Text Available Here, we investigated the effect of chronic mild stress (CMS on the development of atherosclerosis as well as the expression of Toll-like receptors (TLRs signaling pathway in adolescent apolipoprotein E knockout (apoE-/- mice. Mice were subjected to daily CMS for 0, 4, and 12 weeks, respectively. To identify the expression of Toll-like receptor 4 signaling pathway in adolescent apolipoprotein E knockout mice subjected to CMS, we compared gene expression in aortas of stressed and unstressed mice using TLRs signaling pathway real-time PCR microarrays consisting of 87 genes. We found that atherosclerosis lesions both in aortic tress and sinuses of CMS mice were significantly increased linearly in response to duration of CMS exposure. Among 87 genes analyzed, 15 genes were upregulated in stressed mice, especially TLR4, myeloid differentiation factor 88 (MyD88, and IL-1β, and 28 genes were downregulated compared with nonstressed mice. CMS mice demonstrated markedly increased aortic atherosclerosis that were associated with significant increases in levels of expression of TLR4, MyD88, nuclear factor κB (NF-κB, MCP-1, IL-1β, TNF-α, and sICAM-1. Taken together, our results suggest an important role for TLR4 signaling pathway in atherosclerosis in a CMS mouse model.

  7. High resolution mapping of the tropospheric NO2 distribution in three Belgian cities based on airborne APEX remote sensing

    Science.gov (United States)

    Tack, Frederik; Merlaud, Alexis; Fayt, Caroline; Danckaert, Thomas; Iordache, Daniel; Meuleman, Koen; Deutsch, Felix; Adriaenssens, Sandy; Fierens, Frans; Van Roozendael, Michel

    2015-04-01

    An approach is presented to retrieve tropospheric nitrogen dioxide (NO2) vertical column densities (VCDs) and to map the NO2 two dimensional distribution at high resolution, based on Airborne Prism EXperiment (APEX) observations. APEX, developed by a Swiss-Belgian consortium on behalf of ESA (European Space Agency), is a pushbroom hyperspectral imager with a high spatial (approximately 3 m at 5000 m ASL), spectral (413 to 2421 nm in 533 narrow, contiguous spectral bands) and radiometric (14-bit) resolution. VCDs are derived, following a similar approach as described in the pioneering work of Popp et al. (2012), based on (1) spectral calibration and spatial binning of the observed radiance spectra in order to improve the spectral resolution and signal-to-noise ratio, (2) Differential Optical Absorption Spectroscopy (DOAS) analysis of the pre-processed spectra in the visible wavelength region, with a reference spectrum containing low NO2 absorption, in order to quantify the abundance of NO2 along the light path, based on its molecular absorption structures and (3) radiative transfer modeling for air mass factor calculation in order to convert slant to vertical columns. This study will be done in the framework of the BUMBA (Belgian Urban NO2 Monitoring Based on APEX hyperspectral data) project. Dedicated flights with APEX mounted in a Dornier DO-228 airplane, operated by Deutsches Zentrum für Luft- und Raumfahrt (DLR), are planned to be performed in Spring 2015 above the three largest and most heavily polluted Belgian cities, i.e. Brussels, Antwerp and Liège. The retrieved VCDs will be validated by comparison with correlative ground-based and car-based DOAS observations. Main objectives are (1) to assess the operational capabilities of APEX to map the NO2 field over an urban area at high spatial and spectral resolution in a relatively short time and cost-effective way, and to characterise all aspects of the retrieval approach; (2) to use the APEX NO2 measurements

  8. Final-state interactions in two-nucleon knockout reactions

    Science.gov (United States)

    Colle, Camille; Cosyn, Wim; Ryckebusch, Jan

    2016-03-01

    Background: Exclusive two-nucleon knockout after electroexcitation of nuclei [A (e ,e'N N ) in brief] is considered to be a primary source of information about short-range correlations (SRCs) in nuclei. For a proper interpretation of the data, final-state interactions (FSIs) need to be theoretically controlled. Purpose: Our goal is to quantify the role of FSI effects in exclusive A (e ,e'p N ) reactions for four target nuclei representative of the whole mass region. Our focus is on processes that are SRC driven. We investigate the role of FSIs for two characteristic detector setups corresponding to "small" and "large" coverage of the available phase space. Method: Use is made of a factorized expression for the A (e ,e'p N ) cross section that is proportional to the two-body center-of-mass (c.m.) momentum distribution of close-proximity pairs. The A (e ,e'p p ) and A (e ,e'p n ) reactions for the target nuclei 12C,27Al,56Fe, and 208Pb are investigated. The elastic attenuation mechanisms in the FSIs are included using the relativistic multiple-scattering Glauber approximation (RMSGA). Single-charge exchange (SCX) reactions are also included. We introduce the nuclear transparency TAp N, defined as the ratio of exclusive (e ,e'p N ) cross sections on nuclei to those on "free" nucleon pairs, as a measure for the aggregated effect of FSIs in p N knockout reactions from nucleus A . A toy model is introduced in order to gain a better understanding of the A dependence of TAp N. Results: The transparency TAp N drops from 0.2 -0.3 for 12C to 0.04 -0.07 for 208Pb. For all considered kinematics, the mass dependence of TAp N can be captured by the power law TAp N∝A-λ with 0.4 ≲λ ≲0.5 . Apart from an overall reduction factor, we find that FSIs only modestly affect the distinct features of SRC-driven A (e ,e'p N ) which are dictated by the c.m. distribution of close-proximity pairs. Conclusion: The SCX mechanisms represent a relatively small (order of a few percent

  9. Constitutively Expressed αB—Crystallin in Heat Schock Transcription Factor 1 Knockout Mice Myocardium

    Institute of Scientific and Technical Information of China (English)

    刘莉; 张洪慧; 丁国宪; 程蕴琳; 晏良军; BENJAMINIvorJ

    2003-01-01

    Objective:To invesligate the effects of heat shock transcription factor 1(HSF 1) gene on the constitutively expressed aB-Crystallin(αBC)in mice myocardium.Methods:The expression levels of constitutive αBC in HSF1 knockout(hsf1-/-) and HSF1 wild type (hsf1+/+) mice myocardium were evaluated by western blot and immunohistochemistry.Results:The αBC levels in hsf1-/- and hsf1+/_ were 68.42±4.16,100.00±7.58,respectively(P<0.05,cytosolic fraction),and 20.53±1.01,37.55±1.91,respectively(P<0.05,pellet fraction).The αBC signals decreased significantly in hsf1-/- myocardium when compared with those in hsf1+/+ myocardium stained with fluorescence immunohistochemistry.Conclusion.HSF1 is an important,but not the only factor,which mediates the constitutively expressed αBC.

  10. Doublecortin knockout mice show normal hippocampal-dependent memory despite CA3 lamination defects.

    Directory of Open Access Journals (Sweden)

    Johanne Germain

    Full Text Available Mutations in the human X-linked doublecortin gene (DCX cause major neocortical disorganization associated with severe intellectual disability and intractable epilepsy. Although Dcx knockout (KO mice exhibit normal isocortical development and architecture, they show lamination defects of the hippocampal pyramidal cell layer largely restricted to the CA3 region. Dcx-KO mice also exhibit interneuron abnormalities. As well as the interest of testing their general neurocognitive profile, Dcx-KO mice also provide a relatively unique model to assess the effects of a disorganized CA3 region on learning and memory. Based on its prominent anatomical and physiological features, the CA3 region is believed to contribute to rapid encoding of novel information, formation and storage of arbitrary associations, novelty detection, and short-term memory. We report here that Dcx-KO adult males exhibit remarkably preserved hippocampal- and CA3-dependant cognitive processes using a large battery of classical hippocampus related tests such as the Barnes maze, contextual fear conditioning, paired associate learning and object recognition. In addition, we show that hippocampal adult neurogenesis, in terms of proliferation, survival and differentiation of granule cells, is also remarkably preserved in Dcx-KO mice. In contrast, following social deprivation, Dcx-KO mice exhibit impaired social interaction and reduced aggressive behaviors. In addition, Dcx-KO mice show reduced behavioral lateralization. The Dcx-KO model thus reinforces the association of neuropsychiatric behavioral impairments with mouse models of intellectual disability.

  11. The Effect of Different Photoperiods in Circadian Rhythms of Per3 Knockout Mice

    Directory of Open Access Journals (Sweden)

    D. S. Pereira

    2014-01-01

    Full Text Available The aim of this study was to analyse the circadian behavioural responses of mice carrying a functional knockout of the Per3 gene (Per3−/− to different light : dark (L : D cycles. Male adult wild-type (WT and Per3−/− mice were kept under 12-hour light : 12-hour dark conditions (12L : 12D and then transferred to either a short or long photoperiod and subsequently released into total darkness. All mice were exposed to both conditions, and behavioural activity data were acquired through running wheel activity and analysed for circadian characteristics during these conditions. We observed that, during the transition from 12L : 12D to 16L : 8D, Per3−/− mice take approximately one additional day to synchronise to the new L : D cycle compared to WT mice. Under these long photoperiod conditions, Per3−/− mice were more active in the light phase. Our results suggest that Per3−/− mice are less sensitive to light. The data presented here provides further evidence that Per3 is involved in the suppression of behavioural activity in direct response to light.

  12. Hidden disease susceptibility and sexual dimorphism in the heterozygous knockout of Cyp51 from cholesterol synthesis.

    Directory of Open Access Journals (Sweden)

    Monika Lewinska

    Full Text Available We examined the genotype-phenotype interactions of Cyp51+/- mice carrying one functional allele of lanosterol 14α-demethylase from cholesterol biosynthesis. No distinct developmental or morphological abnormalities were observed by routine visual inspection of Cyp51+/- and Cyp51+/+ mice and fertility was similar. We further collected a large data-set from female and male Cyp51+/- mice and controls fed for 16 weeks with three diets and applied linear regression modeling. We used 3 predictor variables (genotype, sex, diet, and 39 response variables corresponding to the organ characteristics (7, plasma parameters (7, and hepatic gene expression (25. We observed significant differences between Cyp51+/- and wild-type mice in organ characteristics and blood lipid profile. Hepatomegaly was observed in Cyp51+/- males, together with elevated total and low-density lipoprotein cholesterol. Cyp51+/- females fed high-fat, high-cholesterol diet were leaner and had elevated plasma corticosterone compared to controls. We observed elevated hepatocyte apoptosis, mitosis and lipid infiltration in heterozygous knockouts of both sexes. The Cyp51+/- females had a modified lipid storage homeostasis protecting them from weight-gain when fed high-fat high-cholesterol diet. Malfunction of one Cyp51 allele therefore initiates disease pathways towards cholesterol-linked liver pathologies and sex-dependent response to dietary challenge.

  13. Improving expression of recombinant human IGF-1 using IGF-1R knockout CHO cell lines.

    Science.gov (United States)

    Romand, Sandrine; Jostock, Thomas; Fornaro, Mara; Schmidt, Joerg; Ritter, Anett; Wilms, Burkhard; Laux, Holger

    2016-05-01

    Chinese Hamster Ovary (CHO) cells are widely used for the large-scale production of recombinant biopharmaceuticals. However, attempts to express IGF-1 (a mutated human Insulin-like growth factor 1 Ea peptide (hIGF-1Ea mut)) in CHO cells resulted in poor cell growth and low productivity (0.1-0.2 g/L). Human IGF-1 variants negatively impacted CHO cell growth via the IGF-1 receptor (IGF-1R). Therefore knockout (KO) of the IGF-1R gene in two different CHO cell lines as well as knockdown (KD) of IGF-1R in one CHO cell line were performed. These cell line engineering approaches decreased significantly the hIGF-1 mediated cell growth inhibition and increased productivity of both KO CHO cell lines as well as of the KD CHO cell line. A productivity increase of 10-fold at pool level and sevenfold at clone level was achieved, resulting in a titer of 1.3 g/L. This data illustrate that cell line engineering approaches are powerful tools to improve the yields of recombinant proteins which are difficult to produce in CHO cells. Biotechnol. Bioeng. 2016;113: 1094-1101. © 2015 Wiley Periodicals, Inc. PMID:26523469

  14. Role of connexin 32 in acetaminophen toxicity in a knockout mice model.

    Science.gov (United States)

    Igarashi, Isao; Maejima, Takanori; Kai, Kiyonori; Arakawa, Shingo; Teranishi, Munehiro; Sanbuissho, Atsushi

    2014-03-01

    Gap junctional intercellular communication (GJIC), by which glutathione (GSH) and inorganic ions are transmitted to neighboring cells, is recognized as being largely involved in toxic processes of chemicals. We examined acetaminophen (APAP)-induced hepatotoxicity clinicopathologically using male wild-type mice and mice lacking the gene for connexin32, a major gap junction protein in the liver [knockout (Cx32KO) mice]. When APAP was intraperitoneally administered at doses of 100, 200, or 300mg/kg, hepatic centrilobular necrosis with elevated plasma aminotransferase activities was observed in wild-type mice receiving 300mg/kg, and in Cx32KO mice given 100mg/kg or more. At 200mg/kg or more, hepatic GSH and GSSG contents decreased significantly and the effect was more severe in wild-type mice than in Cx32KO mice. On the other hand, markedly decreased GSH staining was observed in the hepatic centrilobular zones of Cx32KO mice compared to that of wild-type mice. These results demonstrate that Cx32KO mice are more susceptible to APAP hepatotoxicity than wild-type mice, and indicate that the distribution of GSH of the centrilobular zones in the hepatic lobules, rather than GSH and GSSG contents in the liver, is important in APAP hepatotoxicity. In conclusion, Cx32 protects against APAP-induced hepatic centrilobular necrosis in mice, which may be through the GSH transmission to neighboring hepatocytes by GJIC.

  15. Layer- and column-specific knockout of NMDA receptors in pyramidal neurons of the mouse barrel cortex.

    Directory of Open Access Journals (Sweden)

    Rachel Aronoff

    2007-11-01

    Full Text Available Viral vectors injected into the mouse brain offer the possibility for localized genetic modifications in a highly controlled manner. Lentivector injection into mouse neocortex transduces cells within a diameter of approximately 200µm, which closely matches the lateral scale of a column in barrel cortex. The depth and volume of the injection determines which cortical layer is transduced. Furthermore, transduced gene expression from the lentivector can be limited to predominantly pyramidal neurons by using a 1.3kb fragment of the αCaMKII promoter. This technique therefore allows genetic manipulation of a specific cell type in defined columns and layers of the neocortex. By expressing Cre recombinase from such a lentivector in gene-targeted mice carrying a floxed gene, highly specific genetic lesions can be induced. Here, we demonstrate the utility of this approach by specifically knocking out NMDA receptors (NMDARs in pyramidal neurons in the somatosensory barrel cortex of gene-targeted mice carrying floxed NMDAR 1 genes. Neurons transduced with lentivector encoding GFP and Cre recombinase exhibit not only reductions in NMDAR 1 mRNA levels, but reduced NMDAR-dependent currents and pairing-induced synaptic potentiation. This technique for knockout of NMDARs in a cell type, column- and layer-specific manner in the mouse somatosensory cortex may help further our understanding of the functional roles of NMDARs in vivo during sensory perception and learning.

  16. Mapping the spatio-temporal risk of lead exposure in apex species for more effective mitigation.

    Science.gov (United States)

    Mateo-Tomás, Patricia; Olea, Pedro P; Jiménez-Moreno, María; Camarero, Pablo R; Sánchez-Barbudo, Inés S; Rodríguez Martín-Doimeadios, Rosa C; Mateo, Rafael

    2016-07-27

    Effective mitigation of the risks posed by environmental contaminants for ecosystem integrity and human health requires knowing their sources and spatio-temporal distribution. We analysed the exposure to lead (Pb) in griffon vulture Gyps fulvus-an apex species valuable as biomonitoring sentinel. We determined vultures' lead exposure and its main sources by combining isotope signatures and modelling analyses of 691 bird blood samples collected over 5 years. We made yearlong spatially explicit predictions of the species risk of lead exposure. Our results highlight elevated lead exposure of griffon vultures (i.e. 44.9% of the studied population, approximately 15% of the European, showed lead blood levels more than 200 ng ml(-1)) partly owing to environmental lead (e.g. geological sources). These exposures to environmental lead of geological sources increased in those vultures exposed to point sources (e.g. lead-based ammunition). These spatial models and pollutant risk maps are powerful tools that identify areas of wildlife exposure to potentially harmful sources of lead that could affect ecosystem and human health. PMID:27466455

  17. Cordoma de ápice petroso: relato de um caso Petrous apex chordoma: a case report

    Directory of Open Access Journals (Sweden)

    Cláudio Régis S. Silveira

    2001-02-01

    Full Text Available Cordomas são neoplasias raras que se originam dos remanescentes da notocorda primitiva. Estes remanescentes persistem ao longo de todo o esqueleto axial. Os cordomas intracranianos, mais freqüentemente, se localizam no clivus, próximo à sincondrose esfenooccipital, tipicamente na linha média. Nós descrevemos um caso atípico de cordoma fora da linha média, mais especificamente no ápice petroso, e discutimos as causas embriológicas que determinam esta localização, bem como sintomas, achados de imagem, tratamento cirúrgico e evolução.Chordomas are rare neoplasms arising from notochordal remnants that persist along the axial skeleton. Intracranial chordomas occur more frequently in the midline. We describe an atypical case of an off-midline chordoma arising from the petrous apex, and discuss the embryogenic factors which determine that location, as well as the symptoms, imaging findings, surgical treatment and evolution.

  18. Context Dependent Effect of Landscape on the Occurrence of an Apex Predator across Different Climate Regions.

    Science.gov (United States)

    Fujita, Go; Azuma, Atsuki; Nonaka, Jun; Sakai, Yoshiaki; Sakai, Hatsumi; Iseki, Fumitaka; Itaya, Hiroo; Fukasawa, Keita; Miyashita, Tadashi

    2016-01-01

    In studies of habitat suitability at landscape scales, transferability of species-landscape associations among sites are likely to be critical because it is often impractical to collect datasets across various regions. However, limiting factors, such as prey availability, are not likely to be constant across scales because of the differences in species pools. This is particularly true for top predators that are often the target for conservation concern. Here we focus on gray-faced buzzards, apex predators of farmland-dominated landscapes in East Asia. We investigated context dependency of "buzzard-landscape relationship", using nest location datasets from five sites, each differing in landscape composition. Based on the similarities of prey items and landscape compositions across the sites, we determined several alternative ways of grouping the sites, and then examined whether buzzard-landscape relationship change among groups, which was conducted separately for each way of grouping. As a result, the model of study-sites grouping based on similarities in prey items showed the smallest ΔAICc. Because the terms of interaction between group IDs and areas of broad-leaved forests and grasslands were selected, buzzard-landscape relationship showed a context dependency, i.e., these two landscape elements strengthen the relationship in southern region. The difference in prey fauna, which is associated with the difference in climate, might generate regional differences in the buzzard-landscape associations.

  19. APEX observations of supernova remnants - I. Non-stationary MHD-shocks in W44

    CERN Document Server

    Anderl, S; Güsten, R

    2014-01-01

    Aims. The interaction of supernova remnants (SNRs) with molecular clouds gives rise to strong molecular emission in the far-IR and sub-mm wavelength regimes. The application of MHD shock models in the interpretation of this line emission can yield valuable information on the energetic and chemical impact of supernova remnants. Methods. New mapping observations with the APEX telescope in CO (3-2), (4-3), (6-5), (7-6) and 13CO (3-2) towards two regions in the supernova remnant W44 are presented. Integrated intensities are extracted on five different positions, corresponding to local maxima of CO emission. The integrated intensities are compared to the outputs of a grid of models, which combine an MHD shock code with a radiative transfer module based on the large velocity gradient approximation. Results. All extracted spectra show ambient and line-of-sight components as well as blue- and red-shifted wings indicating the presence of shocked gas. Basing the shock model fits only on the highest-lying transitions th...

  20. A miniature intraventricular axial flow blood pump that is introduced through the left ventricular apex.

    Science.gov (United States)

    Yamazaki, K; Umezu, M; Koyanagi, H; Kitamura, M; Eishi, K; Kawai, A; Tagusari, O; Niinami, H; Akimoto, T; Nojiri, C

    1992-01-01

    A new intraventricular axial flow blood pump has been designed and developed as an implantable left ventricular assist device (LVAD). The pump consists of a tube housing (10 cm in length and 14 mm in diameter), a three-vane impeller combined with a guide vane, and a DC motor. This pump is introduced into the LV cavity through the LV apex, and the outlet cannula is passed antegrade across the aortic valve. Blood is withdrawn from the LV through the inlet ports at the pump base, and discharged into the ascending aorta. A pump flow of > 8 L/min was obtained against 90 mmHg differential pressure in the mock circulatory system. In an acute dog model, this pump could produce a sufficient output of 200 ml/kg/min. In addition, the pump flow profile demonstrated a pulsatile pattern, although the rotation speed was fixed. This is mainly due to the changes in flow rate during a cardiac cycle--that is, during systole, the flow rate increases to the maximum, while the differential pressure between the LV and the aorta decreases to the minimum. Thus, this simple and compact axial flow blood pump can be a potential LVAD, with prompt accessibility and need for less invasive surgical procedures.

  1. Effectiveness of three electronic apex locators to determine root canal working length.

    Directory of Open Access Journals (Sweden)

    Carolina Campillo-Cortes

    2015-08-01

    Full Text Available The aim of this study was to evaluate in vivo the accuracy of three electronic apex locators (EALs in determining working length (WL using hand files and a wear technique. Thirty two premolars that were completely formed apically and that were scheduled for extraction for orthodontic reasons from patients between ages of 15 and 20 years old were included. Electronic measurement of WL was performed using the EAL according to the manufacturer’s instructions. The following three EAL were used: A. Root ZX II; B. Raypex 5, and C. Propex II. There were significant difference (p=0.0002 when comparing median differences among the three EAL. Statistical analysis revealed significant differences between Root ZX II vs. Raypex 5 and Root ZX II vs. Propex II (p=0.0044; p=0.0002, while between Raypex 5 and Propex II, there were no statistically significant differences with respect to the accuracy of the EAL in determining WL (p=0.1087. The present findings suggest that Root ZX II presented the highest agreement rate for determining the final WL.

  2. A COMPARISON BETWEEN SOUNDING DATA AND WRF FORECASTS AT APEX SITE

    Directory of Open Access Journals (Sweden)

    M. Caneo

    2011-01-01

    Full Text Available Cinco configuraciones de WRF usando diferentes modelos de suelo y parametrizaciones de microfísica y de capa límite planetaria se evaluaron con sondeos lanzados durante una campaña de mediciones en el sitio de APEX (Atacama Pathfinder EXperiment. Los resultados indican que los cambios en la parametrización de microfísica no producen cambios apreciables en los perfiles de humedad. El modelo de suelo de Noah muestra menores errores en los perfiles verticales de las variables analizadas en comparación con el esquema de difusión térmica de 5 capas. El análisis de condiciones sin ópticas mostró que las dificultades en predecir la variación diurna en la dirección del viento en condiciones de buen tiempo y la aparición de capas secas poco profundas en la atmósfera son algunas fuentes de errores en los pronósticos.

  3. Twentieth-century decline in streamflows from the hydrographic apex of North America

    Science.gov (United States)

    Rood, Stewart B.; Samuelson, G. M.; Weber, J. K.; Wywrot, K. A.

    2005-05-01

    The Rocky Mountain region near the Canada-United States border provides the North American hydrographic apex with headwater streams flowing to the Pacific, Arctic and Atlantic oceans. The area contains numerous national parks and protected areas with relatively pristine watersheds that permit analyses of historic streamflow patterns with minimal human impacts that would alter hydraulic linkages between precipitation and river discharge. Consequently, we analyzed patterns of mean annual discharge ( Qa) from 31 river reaches that were generally free-flowing with hydrologic records typically commencing in the 1910s and extending to about 2002. To maximize the records of six rivers we undertook regression analyses to extrapolate Qa from sequential hydrometric gauges or from early, summer-only Q data. Spearman ρ and Kendall τ b non-parametric correlations and a parametric approach involving linear regressions combined with analyses of variance were highly consistent in detecting significant historic trends in Qa and the regression analyses estimated the trend magnitudes. These analyses revealed flow declines (exceeding 0.1%/year over the historic record) for 21 reaches (5 with pecoregions.

  4. Chemistry Characterization of Jet Aircraft Engine Particulate by XPS: Results from APEX III

    Science.gov (United States)

    Vander Wal, Randy L.; Bryg, Victoria M.

    2014-01-01

    This paper reports XPS analysis of jet exhaust particulate from a B737, Lear, ERJ, and A300 aircraft during the APEX III NASA led field campaign. Carbon hybridization and bonding chemistry are identified by high-resolution scans about the C1s core-shell region. Significant organic content as gauged by the sp3/sp2 ratio is found across engines and platforms. Polar oxygen functional groups include carboxylic, carbonyl and phenol with combined content of 20 percent or more. By lower resolution survey scans various elements including transition metals are identified along with lighter elements such as S, N, and O in the form of oxides. Burning additives within lubricants are probable sources of Na, Ba, Ca, Zn, P and possibly Sn. Elements present and their percentages varied significantly across all engines, not revealing any trend or identifiable cause for the differences, though the origin is likely the same for the same element when observed. This finding suggests that their presence can be used as a tracer for identifying soots from aircraft engines as well as diagnostic for monitoring engine performance and wear.

  5. Submillimeter Polarimetry with PolKa, a reflection-type modulator for the APEX telescope

    CERN Document Server

    Wiesemeyer, Helmut; Kreysa, Ernst; Weiss, Axel; Güsten, Rolf; Menten, Karl M; Siringo, Giorgio; Schuller, Frederic; Kovacs, Attila

    2014-01-01

    Imaging polarimetry is an important tool for the study of cosmic magnetic fields. In our Galaxy, polarization levels of a few up to 10% are measured in the submillimeter dust emission from molecular clouds and in the synchrotron emission from supernova remnants. Only few techniques exist to image the distribution of polarization angles, tracing the plane-of-sky projection of the magnetic field orientation. At submillimeter wavelengths, polarization is either measured as the differential total power of polarization-sensitive bolometer elements, or by modulating the polarization of the signal. Bolometer arrays such as LABOCA at the APEX telescope are used to observe the continuum emission from fields as large as 0.2 deg. Here we present the results from the commissioning of PolKa, a reflection-type polarimeter for LABOCA. The waveplate has a good efficiency of at least 90%. The modulation efficiency depends mainly on the sampling and on the angular velocity of the waveplate. For the data analysis the concept of...

  6. APEX observations of non-stationary magneto-hydrodynamical shocks in W44

    CERN Document Server

    Anderl, Sibylle; Güsten, Rolf

    2015-01-01

    Supernova remnants (SNRs) are considered as being the sources of galactic cosmic rays. In order to understand the origin, acceleration, and composition of these cosmic rays, detailed knowledge of the physical conditions in the local interstellar medium is needed. The shock interaction of SNRs with molecular clouds that gives rise to strong molecular emission in the far-IR and sub-mm wavelength regimes can be used as a highly valuable tracer of these conditions. The application of MHD shock models in the interpretation of the resulting line emission can yield information on the energetic and chemical impact of supernova remnants. We have mapped two regions in the supernova remnant W44 with the APEX telescope in ${}^{12}$CO (3-2), (4-3), (6-5), (7-6) and ${}^{13}$CO (3-2). The extraction of integrated intensities on five different positions, corresponding to local maxima of CO emission, allows to compare these intensities to the outputs of a grid of models, which combine an MHD shock code with a radiative trans...

  7. Chemistry characterization of jet aircraft engine particulate matter by XPS: Results from APEX III

    Science.gov (United States)

    Vander Wal, Randy L.; Bryg, Victoria M.; Huang, Chung-Hsuan

    2016-09-01

    This paper reports X-ray photoelectron spectroscopy (XPS) analysis of jet exhaust particulate matter (PM) from a B737, Lear, ERJ and A300 aircraft during the APEX III NASA led field campaign. Carbon hybridization and bonding chemistry are identified by high-resolution scans about the C1s core-shell region. Significant organic content as gauged by the sp3/sp2 ratio is found across engines and powers. Polar oxygen functional groups include carboxylic, carbonyl and phenol with combined content of 20% or more. By survey scans various elements including transition metals are identified along with lighter elements such as S, N and O in the form of oxides. Additives within lubricants are probable sources of Na, Ba, Ca, Zn, P and possibly Sn. Elements present and their percentages varied significantly across all engines, not revealing any trend or identifiable cause for the differences, though the origin is likely the same for the same element when observed. This finding suggests that their collective presence could serve as an environmental tracer for identifying PM originating from aircraft engines and serving as a diagnostic for engine performance and wear.

  8. Transient wake and trajectory of free falling cones with various apex angles

    Science.gov (United States)

    Jin, Yaqing; Hamed, Ali M.; Chamorro, Leonardo P.

    2015-11-01

    The early free-fall stages of cones with a density ratio 1.18 and apex angles of 30°, 45°, 60°, and 90° were studied using a wireless 3-axis gyroscope and accelerometer to describe the cone 3D motions, while the induced flow in the near wake was captured using particle image velocimetry. The Reynolds number based on the cone diameter and the velocity at which the cone reaches the first local velocity maximum is found to set the limit between two distinctive states. Before this Re is reached the departure from the vertical path and cone rotations are insignificant, while relatively rapid growth is observed after this Re. Sequences of vertical velocity, swirling strength, LES-decomposed velocity, and pressure fields shows the formation and growth of a large and initially symmetric recirculation bubble at the cone base and highlights the presence of a symmetric 3D vortex rollup dominating the near-wake in the early stages of the fall. Later, the shear layer at the edge of the wake manifests in the shedding of Kelvin-Helmholtz vortices that, due to the nature of the recirculation bubble, reorganize to constitute a part of the rollup. Later in the fall, the wake loses its symmetry and shows a high population of vortical structures leading to turbulence. The asymmetric wake leads to strong interactions between the flow field and the cone creating complex feedback loops.

  9. Abolished synthesis of cholic acid reduces atherosclerotic development in apolipoprotein E knockout mice[S

    OpenAIRE

    Slätis, Katharina; Gåfvels, Mats; Kannisto, Kristina; Ovchinnikova, Olga; Paulsson-Berne, Gabrielle; Parini, Paolo; Jiang, Zhao-Yan; Eggertsen, Gösta

    2010-01-01

    To investigate the effects of abolished cholic acid (CA) synthesis in the ApoE knockout model [apolipoprotein E (apoE) KO],a double-knockout (DKO) mouse model was created by crossbreeding Cyp8b1 knockout mice (Cyp8b1 KO), unable to synthesize the primary bile acid CA, with apoE KO mice. After 5 months of cholesterol feeding, the development of atherosclerotic plaques in the proximal aorta was 50% less in the DKO mice compared with the apoE KO mice. This effect was associated with reduced inte...

  10. CXC receptor knockout mice: characterization of skeletal features and membranous bone healing in the adult mouse.

    Science.gov (United States)

    Bischoff, David S; Sakamoto, Taylor; Ishida, Kenji; Makhijani, Nalini S; Gruber, Helen E; Yamaguchi, Dean T

    2011-02-01

    The potential role of CXC chemokines bearing the glu-leu-arg (ELR) motif in bone repair was studied using a cranial defect (CD) model in mice lacking the CXC receptor (mCXCR(-/-) knockout mice), which is homologous to knockout of the human CXC receptor 2 (CXCR2) gene. During the inflammatory stage of bone repair, ELR CXC chemokines are released by inflammatory cells and serve as chemotactic and angiogenic factors. mCXCR(-/-) mice were smaller in weight and length from base of tail to nose tip, compared to WT littermates. DEXA analysis indicated that bone mineral density (BMD), bone mineral content (BMC), total area (TA), bone area (BA), and total tissue mass (TTM) were decreased in the mCXCR(-/-) mice at 6, 12, and 18 weeks of age. Trabecular bone characteristics in mCXCR(-/-) (% bone, connectivity, number, and thickness) were reduced, and trabecular spacing was increased as evidenced by μCT. There was no difference in bone formation or resorption indices measured by bone histomorphometry. Trabecular BMD was not altered. Cortical bone volume, BMD, and thickness were reduced; whereas, bone marrow volume was increased in mCXCR(-/-). Decreased polar moment of inertia (J) in the tibias/femurs suggested that the mCXCR(-/-) long bones are weaker. This was confirmed by three-point bending testing of the femurs. CDs created in 6-week-old male mCXCR(-/-) and WT littermates were not completely healed at 12 weeks; WT animals, however, had significantly more bone in-growth than mCXCR(-/-). New bone sites were identified using polarized light and assessed for numbers of osteocyte (OCy) lacunae and blood vessels (BlV) around the original CD. In new bone, the number of BlV in WT was >2× that seen in mCXCR(-/-). Bone histomorphometry parameters in the cranial defect did not show any difference in bone formation or resorption markers. In summary, studies showed that mCXCR(-/-) mice have (1) reduced weight and size; (2) decreased BMD and BMC; (3) decreased amounts of trabecular

  11. Exposure to low-dose rotenone precipitates synaptic plasticity alterations in PINK1 heterozygous knockout mice.

    Science.gov (United States)

    Martella, G; Madeo, G; Maltese, M; Vanni, V; Puglisi, F; Ferraro, E; Schirinzi, T; Valente, E M; Bonanni, L; Shen, J; Mandolesi, G; Mercuri, N B; Bonsi, P; Pisani, A

    2016-07-01

    Heterozygous mutations in the PINK1 gene are considered a susceptibility factor to develop early-onset Parkinson's disease (PD), as supported by dopamine hypometabolism in asymptomatic mutation carriers and subtle alterations of dopamine-dependent striatal synaptic plasticity in heterozygous PINK1 knockout (PINK1(+/-)) mice. The aim of the present study was to investigate whether exposure to low-dose rotenone of heterozygous PINK1(+/-) mice, compared to their wild-type PINK1(+/+) littermates, could impact on dopamine-dependent striatal synaptic plasticity, in the absence of apparent structural alterations. Mice were exposed to a range of concentrations of rotenone (0.01-1mg/kg). Chronic treatment with concentrations of rotenone up to 0.8mg/kg did not cause manifest neuronal loss or changes in ATP levels both in the striatum or substantia nigra of PINK1(+/-) and PINK1(+/+) mice. Moreover, rotenone (up to 0.8mg/kg) treatment did not induce mislocalization of the mitochondrial membrane protein Tom20 and release of cytochrome c in PINK1(+/-) striata. Accordingly, basic electrophysiological properties of nigral dopaminergic and striatal medium spiny neurons (MSNs) were normal. Despite the lack of gross alterations in neuronal viability in chronically-treated PINK1(+/-), a complete loss of both long-term depression (LTD) and long-term potentiation (LTP) was recorded in MSNs from PINK1(+/-) mice treated with a low rotenone (0.1mg/kg) concentration. Even lower concentrations (0.01mg/kg) blocked LTP induction in heterozygous PINK1(+/-) MSNs compared to PINK1(+/+) mice. Of interest, chronic pretreatment with the antioxidants alpha-tocopherol and Trolox, a water-soluble analog of vitamin E and powerful antioxidant, rescued synaptic plasticity impairment, confirming that, at the doses we utilized, rotenone did not induce irreversible alterations. In this model, chronic exposure to low-doses of rotenone was not sufficient to alter mitochondrial integrity and ATP production, but

  12. Drop tests of the Three Mile Island knockout canister

    International Nuclear Information System (INIS)

    A type of Three Mile Island Unit 2 (TMI-2) defueling canister, called a ''knockout'' canister, was subjected to a series of drop tests at the Oak Ridge National Laboratory's Drop Test Facility. These tests were designed to confirm the structural integrity of internal fixed neutron poisons in support of a request for NRC licensing of this type of canister for the shipment of TMI-2 reactor fuel debris to the Idaho National Engineering Laboratory (INEL) for the Core Examination R and D Program. Work conducted at the Oak Ridge National Laboratory included (1) precise physical measurements of the internal poison rod configuration before assembly, (2) canister assembly and welding, (3) nondestructive examination (an initial hydrostatic pressure test and an x-ray profile of the internals before and after each drop test), (4) addition of a simulated fuel load, (5) instrumentation of the canister for each drop test, (6) fabrication of a cask simulation vessel with a developed and tested foam impact limiter, (7) use of refrigeration facilities to cool the canister to well below freezing prior to three of the drops, (8) recording the drop test with still, high-speed, and normal-speed photography, (9) recording the accelerometer measurements during impact, (10) disassembly and post-test examination with precise physical measurements, and (11) preparation of the final report

  13. Final-state interactions in two-nucleon knockout reactions

    CERN Document Server

    Colle, Camille; Ryckebusch, Jan

    2015-01-01

    Background: Exclusive two-nucleon knockout after electroexcitation of nuclei ($A(e,e'NN)$ in brief) is considered to be a primary source of information about short-range correlations (SRC) in nuclei. For a proper interpretation of the data, final-state interactions (FSI) need to be theoretically controlled. Purpose: Our goal is to quantify the role of FSI effects in exclusive $A(e,e'pN)$ reactions for four target nuclei representative for the whole mass region. Our focus is on processes that are SRC driven. We investigate the role of FSI for two characteristic detector setups corresponding with a "small" and "large" coverage of the available phase space. Results: The transparency $T^{pN}_{A}$, defined as the ratio of exclusive $(e,e'pN)$ cross sections on nuclei to those on "free" nucleon pairs, drops from $ 0.2-0.3 $ for $^{12}$C to $0.04-0.07$ for $^{208}$Pb. For all considered kinematics, the mass dependence of the $T^{pN}_{A}$ can be captured by the power law $T^{pN}_{A} \\propto A^{- \\lambda}$ with $ 0.4 ...

  14. Geometric analysis of Arabidopsis root apex reveals a new aspect of the ethylene signal transduction pathway in development

    Science.gov (United States)

    Cervantes, Emilio; Tocino, Angel

    2005-01-01

    Structurally, ethylene is the simplest phytohormone and regulates multiple aspects of plant growth and development. Its effects are mediated by a signal transduction cascade involving receptors, MAP kinases and transcription factors. Many morphological effects of ethylene in plant development, including root size, have been previously described. In this article a combined geometric and algebraic approach has been used to analyse the shape and the curvature in the root apex of Arabidopsis seedlings. The process requires the fitting of Bezier curves that reproduce the root apex shape, and the calculation of the corresponding curvatures. The application of the method has allowed us to identify significant differences in the root curvatures of ethylene insensitive mutants (ein2-1 and etr1-1) with respect to the wild-type Columbia.

  15. Comparative evaluation of accuracy of two electronic apex locators in the presence of various irrigants: An in vitro study

    Directory of Open Access Journals (Sweden)

    Saru Jain

    2012-01-01

    Full Text Available Context: The establishment of appropriate working length is one of the most critical steps in endodontic therapy. Electronic apex locators have been introduced to determine the working length. The development of electronic apex locators has helped make the assessment of the working length more accurate and predictable, along with reduction in treatment time and radiation dose. Objectives: The aim of this study was to compare the efficacy of electronic apex locators after cleansing and shaping of the root canals and whether there was any alteration in accuracy when used in the presence of irrigants. Materials and Methods: Seventy extracted human permanent molars with mature apices were selected. Equal number of maxillary and mandibular permanent molars (35 each were sectioned at the cemento-enamel junction. Access opening was done and only the mesiobuccal root canal was studied for the purpose of standardization. Electronic working length measurements were taken before and after preparation of the mesiobuccal canal with Root ZX and ProPex II using various irrigants. Statistical Analysis Used: The data were statistically analyzed using a paired t-test at 0.05 level of significance. Results: P-values for actual and final canal lengths for Root ZX employing NaoCl(0.001, CHX(0.006, LA(0.020 and for ProPex II was (0.001 respectively. When the data were compared, results were statistically significant (P < 0.05. Conclusion: Within the limitations of this study Root ZX can be considered to be an accurate electronic apex locator and CHX as irrigant matched more precisely with the actual canal length measurements.

  16. Non-surgical management of a mutilated maxillary central incisor with open apex and large periapical lesion

    OpenAIRE

    Puneet Ajwani; Nalini Saini

    2011-01-01

    A 24-year-old female patient reported with a mutilated maxillary left central incisor. The coronal tooth structure remaining was very less, discolored, and brittle. She gave history of trauma about 15 years back when the tooth got fractured. An intraoral periapical radiograph revealed an open apex and a large periapical lesion. The case was managed successfully by conservative means using intracanal calcium hydroxide and mineral trioxide aggregate (MTA) apical barrier followed by a fiber post...

  17. Clinical Comparison of Root Length Measurements with Electronic Apex Locator and Conventional Radiography in Mandibular Deciduous Teeth

    OpenAIRE

    Eskandarian T; Sahebi S.; Amirabadi F.; Alipour A

    2011-01-01

    Statement of Problem: Success in pulpectomy of deciduous teeth greatly depends on the accuracy of root length measurements which have mostly been done radiographically. However, X-ray risks and patient cooperation have usually been a challenge for the clinicians.Purpose: The clinical comparison of root length measurements with electronic apex locator and conventional radiography in 4-6 year old children's mandibular deciduous teeth was the aim of the present study.Materials and Method: In the...

  18. Comparison of digital radiography and apex locator with the conventional method in root length determination of primary teeth

    OpenAIRE

    I E Neena; A Ananthraj; P. Praveen; Karthik, V.; P. Rani

    2011-01-01

    Aim: The purpose of this study was to compare the Working length in primary teeth endodontics using intra oral digital radiovisiography and apex locator with conventional method for accuracy. Materials and Methods: This in vivo study was conducted on 30 primary teeth which were indicated for pulpectomy in the patients of the age group of 5-11 years All experimental teeth had adequate remaining tooth structure for rubber dam isolation and radiographicaly visible canals. Endodontic treatment wa...

  19. White Sharks (Carcharodon carcharias) Scavenging on Whales and Its Potential Role in Further Shaping the Ecology of an Apex Predator

    OpenAIRE

    Chris Fallows; Gallagher, Austin J.; Neil Hammerschlag

    2013-01-01

    Scavenging, a result of a temporary pulse of resources, occurs in virtually all ecosystems containing carnivores, and is an important energy transfer pathway that can impact ecosystem structure and function, and this ecological significance has largely been considered from a terrestrial standpoint; however, little is known about the role of scavenging in shaping the behavioral ecology of marine species, specifically apex predators. Here we present findings from multiple opportunistic observat...

  20. [Injuries of the pleural cupula and lung apex during catheterization of the subclavian vein, and their prevention].

    Science.gov (United States)

    Barskiĭ, A V; Makharadze, N L

    1983-03-01

    In the clinic 1442 catheterizations were performed, 1156 of them being catheterizations of the subclavian vein. Dangerous complications such as punctures of the posterior wall of the subclavian vein in combination with injuries of the pleura cupola and apex pulmonis were observed three times. Hemo-, hydro- and pneumothorax accompanying these complications in different combinations are contraindications to repeated puncture and catheterization of the subclavian vein both in the left and in the right. Recommendations are given to reduced possible complications.