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Sample records for apex gene knockout

  1. The biology of novel animal genes: Mouse APEX gene knockout

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    MacInnes, M.; Altherr, M.R.; Ludwig, D. [Los Alamos National Lab., NM (United States); Pedersen, R.; Mold, C. [Univ. of California, San Francisco, CA (United States)

    1997-07-01

    This is the final report of a one-year, Laboratory Directed Research and Development (LDRD) project at the Los Alamos National Laboratory (LANL). The controlled breeding of novel genes into mice, including the gene knockout (KO), or conversely by adding back transgenes provide powerful genetic technologies that together suffice to determine in large part the biological role(s) of novel genes. Inbred mouse remains the best understood and most useful mammalian experimental system available for tackling the biology of novel genes. The major mammalian apurinic/apyrimidinic (AP) endonuclease (APE), is involved in a key step in the repair of spontaneous and induced AP sites in DNA. Efficient repair of these lesions is imperative to prevent the stable incorporation of mutations into the cellular genome which may lead to cell death or transformation. Loss or modulation of base excison repair activity in vivo may elevate the spontaneous mutation rate in cells, and may lead to a substantial increase in the incidence of cancer. Despite extensive biochemical analysis, however, the significance of these individual APE functions in vivo has not been elucidated. Mouse embryonic stem (ES) cells heterozygous for a deletion mutation in APE have been generated and whole animals containing the APE mutation have been derived from these ES cells. Animals homozygous for the APE null mutation die early in gestation, underscoring the biological significance of this DNA repair gene.

  2. New findings in gene knockout, mutant and transgenic mice.

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    Bartke, Andrzej

    2008-01-01

    During the past year, some novel genetic modifications were shown to alter the lifespan of mice, thus expanding the list of genes and physiological processes that influence mammalian aging. Considerable progress was also made in identifying putative mechanisms of extended longevity in previously described gene knockouts, mutants and transgenics. In addition, new leads concerning mechanisms of aging were derived from studies of gene knockout mice in which aging is accelerated. Among the important findings from the period July 2006 to July 2007: Core body temperature was shown to influence longevity in homeothermic animals; a Surf1 gene knockout extended lifespan in mice; separate studies using Little and Snell dwarf mice found stress resistance enhancements correlated with longevity gains; and mice heterozygous for deletion of insulin receptor substrate 2 (IRS-2) lived longer than normal animals, while animals with homozygous or heterozygous deletion of IRS-2 selectively in the brain exhibited comparable extension of lifespan and various symptoms of delayed aging. PMID:18053667

  3. 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

  4. 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...

  5. Reduced body weight is a common effect of gene knockout in mice

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    Lawler Maureen P

    2008-01-01

    Full Text Available Abstract Background During a search for obesity candidate genes in a small region of the mouse genome, we noticed that many genes when knocked out influence body weight. To determine whether this was a general feature of gene knockout or a chance occurrence, we surveyed the Jackson Laboratory Mouse Genome Database for knockout mouse strains and their phenotypes. Body weights were not available for all strains so we also obtained body weight information by contacting a random sample of investigators responsible for a knockout strain. Results We classified each knockout mouse strain as (1 lighter and smaller, (2 larger and heavier, or (3 the same weight, relative to control mice. We excluded knockout strains that died early in life, even though this type of lethality is often associated with a small embryo or reduced body size. Based on a dataset of 1,977 knockout strains, we found that that 31% of viable knockout mouse strains weighed less and an additional 3% weighed more than did controls. Conclusion Body weight is potentially a latent variable in about a third of experiments that use knockout mice and should be considered in interpreting experimental outcomes, e.g., in studies of hypertension, drug and hormone metabolism, organ development, cell proliferation and apoptosis, digestion, heart rate, or atherosclerosis. If we assume that the knockout genes we surveyed are representative then upward of 6,000 genes are predicted to influence the size of a mouse. Body weight is highly heritable, and numerous quantitative trait loci have been mapped in mice, but "multigenic" is an insufficient term for the thousands of loci that could contribute to this complex trait.

  6. 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 ...

  7. Characterization of physiological responses to 22 gene knockouts in Escherichia coli central carbon metabolism.

    Science.gov (United States)

    Long, Christopher P; Gonzalez, Jacqueline E; Sandoval, Nicholas R; Antoniewicz, Maciek R

    2016-09-01

    Understanding the impact of gene knockouts on cellular physiology, and metabolism in particular, is centrally important to quantitative systems biology and metabolic engineering. Here, we present a comprehensive physiological characterization of wild-type Escherichia coli and 22 knockouts of enzymes in the upper part of central carbon metabolism, including the PTS system, glycolysis, pentose phosphate pathway and Entner-Doudoroff pathway. Our results reveal significant metabolic changes that are affected by specific gene knockouts. Analysis of collective trends and correlations in the data using principal component analysis (PCA) provide new, and sometimes surprising, insights into E. coli physiology. Additionally, by comparing the data-to-model predictions from constraint-based approaches such as FBA, MOMA and RELATCH we demonstrate the important role of less well-understood kinetic and regulatory effects in central carbon metabolism. PMID:27212692

  8. 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

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

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    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

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

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    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

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

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    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.

  12. 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.

  13. Determination of antibiotic hypersensitivity among 4,000 single-gene-knockout mutants of Escherichia coli.

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    Tamae, Cindy; Liu, Anne; Kim, Katherine; Sitz, Daniel; Hong, Jeeyoon; Becket, Elinne; Bui, Ann; Solaimani, Parrisa; Tran, Katherine P; Yang, Hanjing; Miller, Jeffrey H

    2008-09-01

    We have tested the entire Keio collection of close to 4,000 single-gene knockouts in Escherichia coli for increased susceptibility to one of seven different antibiotics (ciprofloxacin, rifampin, vancomycin, ampicillin, sulfamethoxazole, gentamicin, or metronidazole). We used high-throughput screening of several subinhibitory concentrations of each antibiotic and reduced more than 65,000 data points to a set of 140 strains that display significantly increased sensitivities to at least one of the antibiotics, determining the MIC in each case. These data provide targets for the design of "codrugs" that can potentiate existing antibiotics. We have made a number of double mutants with greatly increased sensitivity to ciprofloxacin, and these overcome the resistance generated by certain gyrA mutations. Many of the gene knockouts in E. coli are hypersensitive to more than one antibiotic. Together, all of these data allow us to outline the cell's "intrinsic resistome," which provides innate resistance to antibiotics. PMID:18621901

  14. CRISPR/Cas9-mediated gene knockout in the mouse brain using in utero electroporation

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    Shinmyo, Yohei; Tanaka, Satoshi; Tsunoda, Shinichi; Hosomichi, Kazuyoshi; Tajima, Atsushi; Kawasaki, Hiroshi

    2016-01-01

    The CRISPR/Cas9 system has recently been adapted for generating knockout mice to investigate physiological functions and pathological mechanisms. Here, we report a highly efficient procedure for brain-specific disruption of genes of interest in vivo. We constructed pX330 plasmids expressing humanized Cas9 and single-guide RNAs (sgRNAs) against the Satb2 gene, which encodes an AT-rich DNA-binding transcription factor and is responsible for callosal axon projections in the developing mouse brain. We first confirmed that these constructs efficiently induced double-strand breaks (DSBs) in target sites of exogenous plasmids both in vitro and in vivo. We then found that the introduction of pX330-Satb2 into the developing mouse brain using in utero electroporation led to a dramatic reduction of Satb2 expression in the transfected cerebral cortex, suggesting DSBs had occurred in the Satb2 gene with high efficiency. Furthermore, we found that Cas9-mediated targeting of the Satb2 gene induced abnormalities in axonal projection patterns, which is consistent with the phenotypes previously observed in Satb2 mutant mice. Introduction of pX330-NeuN using our procedure also resulted in the efficient disruption of the NeuN gene. Thus, our procedure combining the CRISPR/Cas9 system and in utero electroporation is an effective and rapid approach to achieve brain-specific gene knockout in vivo. PMID:26857612

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

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    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

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

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    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.

  17. 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.

  18. Establishment of large mutant families of tomato for gene knockouts and other important traits

    International Nuclear Information System (INIS)

    Tomato (Lycopersicon esculentum L.) is one of the popular and important vegetable crops grown worldwide. It ranks second after potato in terms of global production area. Tomato is the most important crop in the fresh and processed vegetable market. Current breeding efforts are geared towards the incorporation of disease resistance genes, enhanced quality traits and other important traits required by the tomato crop to sustain productivity under biotic and abiotic limiting conditions. As sources for genetic stocks, breeding materials are resourced from within the Lycopersicon and wild relatives. This paper reports the successful establishment of large M1 and M2 families of tomato generated using physical (Cobalt 60 gamma ray) and chemical (ethylmethane sulfonate, EMS) mutagens. The mutant germplasm will be used as a rich source of genetic materials to intensify crop improvement and genetic studies in tomato. Based on high-throughput phenotype screening, a total of forty one (41) homogeneous and segregating M2 families were identified as visible mutants. The most common visible mutations observed in the M2 screening were the monopodial plant type, different forms of chlorotic mutants, and plants with abnormal leaf morphology. From the large 600Gy and 1.0% EMS mutant families, 12 families were also identified as initial bacterial wilt resistance (BWR) gene knockouts. More gene knockouts, and visible and biochemical mutations will be identified from the remaining 600Gy and 1.0% EMS M2 families. To confirm mutation, targeted screening will be employed using gene-specific DNA markers like BWR SCAR markers and published EST-derived markers for tomato mutant genes. This is to determine and compare the type/frequency of mutations that have been induced using either gamma ray irradiation or EMS. (author)

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

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    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

  20. 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.

  1. Pol II-expressed shRNA knocks down Sod2 gene expression and causes phenotypes of the gene knockout in mice.

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    2006-01-01

    Full Text Available RNA interference (RNAi has been used increasingly for reverse genetics in invertebrates and mammalian cells, and has the potential to become an alternative to gene knockout technology in mammals. Thus far, only RNA polymerase III (Pol III-expressed short hairpin RNA (shRNA has been used to make shRNA-expressing transgenic mice. However, widespread knockdown and induction of phenotypes of gene knockout in postnatal mice have not been demonstrated. Previous studies have shown that Pol II synthesizes micro RNAs (miRNAs-the endogenous shRNAs that carry out gene silencing function. To achieve efficient gene knockdown in mammals and to generate phenotypes of gene knockout, we designed a construct in which a Pol II (ubiquitin C promoter drove the expression of an shRNA with a structure that mimics human miRNA miR-30a. Two transgenic lines showed widespread and sustained shRNA expression, and efficient knockdown of the target gene Sod2. These mice were viable but with phenotypes of SOD2 deficiency. Bigenic heterozygous mice generated by crossing these two lines showed nearly undetectable target gene expression and phenotypes consistent with the target gene knockout, including slow growth, fatty liver, dilated cardiomyopathy, and premature death. This approach opens the door of RNAi to a wide array of well-established Pol II transgenic strategies and offers a technically simpler, cheaper, and quicker alternative to gene knockout by homologous recombination for reverse genetics in mice and other mammalian species.

  2. 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

  3. 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

  4. 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

  5. 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

  6. Determination of hypersensitivity to genotoxic agents among Escherichia coli single gene knockout mutants.

    Science.gov (United States)

    Becket, Elinne; Chen, Frank; Tamae, Cindy; Miller, Jeffrey H

    2010-09-01

    We have tested the KEIO collection of 3985 different viable single gene knockouts in Escherichia coli to identify genes whose loss increases sensitivity to one or more of six different chemotherapeutic agents and mutagens: Bleomycin (BLM), Cisplatin (CPT), ICR-191 (ICR), 5-azacytidine (5AZ), Zebularine (ZEB), and 5-bromo-2'-deoxyuridine (5BdU). We discovered a set of 156 strains that display a significant increase in sensitivity to at least one of the agents tested. Each genotoxic agent generates a distinct "sensitivity profile" that is characteristic of the agent. Comparison with an independent study of sensitivity profiles for an extensive set of antibiotics pinpoints those effects that are relatively specific for each agent. In some cases engineered double mutants have greatly increased effects. These results provide insight into the mechanism of action of each agent, and define targets for the design of co-drugs that can potentiate these agents. An example is the finding that mutants lacking one of several genes in the folate biosynthetic pathway are hypersensitive to ZEB, leading to a demonstration of synergy between trimethoprim and ZEB. PMID:20674514

  7. 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

  8. 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...

  9. 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.

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

    Science.gov (United States)

    Zuo, Qisheng; Wang, Yinjie; Cheng, Shaoze; Lian, Chao; Tang, Beibei; Wang, Fei; Lu, Zhenyu; Ji, Yanqing; Zhao, Ruifeng; Zhang, Wenhui; Jin, Kai; Song, Jiuzhou; Zhang, Yani; Li, Bichun

    2016-01-01

    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. PMID:27172204

  11. 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.

  12. 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.

  13. Antibiotic sensitivity profiles determined with an Escherichia coli gene knockout collection: generating an antibiotic bar code.

    Science.gov (United States)

    Liu, Anne; Tran, Lillian; Becket, Elinne; Lee, Kim; Chinn, Laney; Park, Eunice; Tran, Katherine; Miller, Jeffrey H

    2010-04-01

    We have defined a sensitivity profile for 22 antibiotics by extending previous work testing the entire KEIO collection of close to 4,000 single-gene knockouts in Escherichia coli for increased susceptibility to 1 of 14 different antibiotics (ciprofloxacin, rifampin [rifampicin], vancomycin, ampicillin, sulfamethoxazole, gentamicin, metronidazole, streptomycin, fusidic acid, tetracycline, chloramphenicol, nitrofurantoin, erythromycin, and triclosan). We screened one or more subinhibitory concentrations of each antibiotic, generating more than 80,000 data points and allowing a reduction of the entire collection to a set of 283 strains that display significantly increased sensitivity to at least one of the antibiotics. We used this reduced set of strains to determine a profile for eight additional antibiotics (spectinomycin, cephradine, aztreonem, colistin, neomycin, enoxacin, tobramycin, and cefoxitin). The profiles for the 22 antibiotics represent a growing catalog of sensitivity fingerprints that can be separated into two components, multidrug-resistant mutants and those mutants that confer relatively specific sensitivity to the antibiotic or type of antibiotic tested. The latter group can be represented by a set of 20 to 60 strains that can be used for the rapid typing of antibiotics by generating a virtual bar code readout of the specific sensitivities. Taken together, these data reveal the complexity of intrinsic resistance and provide additional targets for the design of codrugs (or combinations of drugs) that potentiate existing antibiotics. PMID:20065048

  14. Kidney development and gene expression in the HIF2alpha knockout mouse.

    Science.gov (United States)

    Steenhard, Brooke M; Freeburg, Paul B; Isom, Kathryn; Stroganova, Larysa; Borza, Dorin-Bogdan; Hudson, Billy G; St John, Patricia L; Zelenchuk, Adrian; Abrahamson, Dale R

    2007-04-01

    The hypoxia-inducible transcription factor-2 (HIF2), a heterodimer composed of HIF2alpha and HIF1beta subunits, drives expression of genes essential for vascularization, including vascular endothelial growth factor (VEGF) and VEGF receptor-2 (VEGFR-2, Flk-1). Here, we used a HIF2alpha/LacZ transgenic mouse to define patterns of HIF2alpha transcription during kidney development and maturation. Our results from embryonic heterozygotes showed HIF2alpha/LacZ expression by apparently all renal endothelial cells. At 4 weeks of age, glomerular mesangial and vascular smooth muscle cells were also positive together with endothelial cells. These expression patterns were confirmed by electron microscopy using Bluo-gal as a beta-galactosidase substrate. Small numbers of glomerular and tubular epithelial cells were also positive at all stages examined. Light and electron microscopic examination of kidneys from HIF2alpha null embryos showed no defects in renal vascular development or nephrogenesis. Similarly, the same amounts of Flk-1 protein were seen on Western blots of kidney extracts from homozygous and heterozygous HIF2alpha mutants. To examine responsiveness of HIF2alpha null kidneys to hypoxia, embryonic day 13.5 metanephroi were cultured in room air or in mild (5% O(2)) hypoxia. For both heterozygous and null samples, VEGF mRNA levels doubled when metanephroi were cultured in mild hypoxia. Anterior chamber grafts of embryonic HIF2alpha knockouts were morphologically indistinguishable from heterozygous grafts. Endothelial markers, platelet endothelial cell adhesion molecule and BsLB4, as well as glomerular epithelial markers, GLEPP1 and WT-1, were all expressed appropriately. Finally, we undertook quantitative real-time polymerase chain reaction of kidneys from HIF2alpha null embryos and wild-type siblings and found no compensatory up-regulation of HIF1alpha or -3alpha. Our results show that, although HIF2alpha was widely transcribed by kidney endothelium and vascular

  15. 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 < 0.05). However, these bone microstructural parameters were increased by 53%, 34% and 40%, respectively, in IL6(-/-) mice than those in WT mice on the HFD (P < 0.05). IL6(-/-) osteoblasts displayed higher 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

  16. Gene knockout of the intracellular amylase gene by homologous recombination in Streptococcus bovis.

    Science.gov (United States)

    Brooker, J D; McCarthy, J M

    1997-09-01

    Streptococcus bovis expresses two different amylases, one intracellular and the other secreted. A suicide vector containing part of the intracellular alpha-amylase gene from Streptococcus bovis WI-1 was recombined into the S. bovis WI-1 chromosome to disrupt the endogenous gene. Recombination was demonstrated by Southern blot, and zymogram analysis confirmed the loss of the intracellular amylase. Amylase activity in cell-free extracts of the recombinant grown in the presence of 1% starch was only 7% of wild type. The rate of logarithmic growth of the recombinant was 15-20% of the wild type in medium containing either 1% glucose, starch, or cellobiose. Revertants and non-amylase control recombinants had logarithmic growth rates that were the same as wild type. Plasmid transformants containing multiple copies of the cloned gene expressed up to threefold higher levels of intracellular amylase activity than wild type but did not demonstrate elevated growth rates. These results suggest that a critical level of expression of the intracellular amylase gene may be important for rapid growth of the bacterium. PMID:9236293

  17. 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...

  18. 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.

  19. 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

  20. 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

  1. 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.

  2. 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.

  3. 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

  4. 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...

  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. Modeling RNA polymerase competition: the effect of σ-subunit knockout and heat shock on gene transcription level

    Directory of Open Access Journals (Sweden)

    Seliverstov Alexandr V

    2011-01-01

    Full Text Available Abstract Background Modeling of a complex biological process can explain the results of experimental studies and help predict its characteristics. Among such processes is transcription in the presence of competing RNA polymerases. This process involves RNA polymerases collision followed by transcription termination. Results A mathematical and computer simulation model is developed to describe the competition of RNA polymerases during genes transcription on complementary DNA strands. E.g., in the barley Hordeum vulgare the polymerase competition occurs in the locus containing plastome genes psbA, rpl23, rpl2 and four bacterial type promoters. In heat shock experiments on isolated chloroplasts, a twofold decrease of psbA transcripts and even larger increase of rpl23-rpl2 transcripts were observed, which is well reproduced in the model. The model predictions are in good agreement with virtually all relevant experimental data (knockout, heat shock, chromatogram data, etc.. The model allows to hypothesize a mechanism of cell response to knockout and heat shock, as well as a mechanism of gene expression regulation in presence of RNA polymerase competition. The model is implemented for multiprocessor platforms with MPI and supported on Linux and MS Windows. The source code written in C++ is available under the GNU General Public License from the laboratory website. A user-friendly GUI version is also provided at http://lab6.iitp.ru/en/rivals. Conclusions The developed model is in good agreement with virtually all relevant experimental data. The model can be applied to estimate intensities of binding of the holoenzyme and phage type RNA polymerase to their promoters using data on gene transcription levels, as well as to predict characteristics of RNA polymerases and the transcription process that are difficult to measure directly, e.g., the intensity (frequency of holoenzyme binding to the promoter in correlation to its nucleotide composition and the

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

    Directory of Open Access Journals (Sweden)

    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.

  8. 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.

  9. 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.

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

    Science.gov (United States)

    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

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

    Directory of Open Access Journals (Sweden)

    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. Orexin gene transfer into the amygdala suppresses both spontaneous and emotion-induced cataplexy in orexin-knockout mice.

    Science.gov (United States)

    Liu, Meng; Blanco-Centurion, Carlos; Konadhode, Roda Rani; Luan, Liju; Shiromani, Priyattam J

    2016-03-01

    Narcolepsy is a chronic sleep disorder linked to the loss of orexin-producing neurons in the hypothalamus. Cataplexy, a sudden loss of muscle tone during waking, is an important distinguishing symptom of narcolepsy and it is often triggered by strong emotions. The neural circuit underlying cataplexy attacks is not known, but is likely to involve the amygdala, a region implicated in regulating emotions. In mice models of narcolepsy, transfer of the orexin gene into surrogate neurons has been successful in ameliorating narcoleptic symptoms. However, it is not known whether this method also blocks cataplexy triggered by strong emotions. To examine this possibility, the gene encoding mouse prepro-orexin was transferred into amygdala neurons of orexin-knockout (KO) mice (rAAV-orexin; n = 8). Orexin-KO mice that did not receive gene transfer (no-rAAV; n = 7) or received only the reporter gene (rAAV-GFP; n = 7) served as controls. Three weeks later, the animal's sleep and behaviour were recorded at night (no-odour control night), followed by another recording at night in the presence of predator odour (odour night). Orexin-KO mice given the orexin gene transfer into surrogate amygdala neurons had significantly less spontaneous bouts of cataplexy, and predator odour did not induce cataplexy compared with control mice. Moreover, the mice with orexin gene transfer were awake more during the odour night. These results demonstrate that orexin gene transfer into amygdala neurons can suppress both spontaneous and emotion-induced cataplexy attacks in narcoleptic mice. It suggests that manipulating amygdala pathways is a potential strategy for treating cataplexy in narcolepsy. PMID:26741960

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

    Science.gov (United States)

    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

  14. 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

  15. PARP1 gene knock-out increases resistance to retinal degeneration without affecting retinal function.

    Directory of Open Access Journals (Sweden)

    Ayse Sahaboglu

    Full Text Available Retinitis pigmentosa (RP is a group of inherited neurodegenerative diseases affecting photoreceptors and causing blindness in humans. Previously, excessive activation of enzymes belonging to the poly-ADP-ribose polymerase (PARP group was shown to be involved in photoreceptor degeneration in the human homologous rd1 mouse model for RP. Since there are at least 16 different PARP isoforms, we investigated the exact relevance of the predominant isoform - PARP1 - for photoreceptor cell death using PARP1 knock-out (KO mice. In vivo and ex vivo morphological analysis using optic coherence tomography (OCT and conventional histology revealed no major alterations of retinal phenotype when compared to wild-type (wt. Likewise, retinal function as assessed by electroretinography (ERG was normal in PARP1 KO animals. We then used retinal explant cultures derived from wt, rd1, and PARP1 KO animals to test their susceptibility to chemically induced photoreceptor degeneration. Since photoreceptor degeneration in the rd1 retina is triggered by a loss-of-function in phosphodiesterase-6 (PDE6, we used selective PDE6 inhibition to emulate the rd1 situation on non-rd1 genotypes. While wt retina subjected to PDE6 inhibition showed massive photoreceptor degeneration comparable to rd1 retina, in the PARP1 KO situation, cell death was robustly reduced. Together, these findings demonstrate that PARP1 activity is in principle dispensable for normal retinal function, but is of major importance for photoreceptor degeneration under pathological conditions. Moreover, our results suggest that PARP dependent cell death or PARthanatos may play a major role in retinal degeneration and highlight the possibility to use specific PARP inhibitors for the treatment of RP.

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

    Science.gov (United States)

    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

  17. 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)

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

    Directory of Open Access Journals (Sweden)

    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

  19. 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

  20. 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

  1. 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

  2. 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

  3. Antibiotic Sensitivity Profiles Determined with an Escherichia coli Gene Knockout Collection: Generating an Antibiotic Bar Code ▿ †

    Science.gov (United States)

    Liu, Anne; Tran, Lillian; Becket, Elinne; Lee, Kim; Chinn, Laney; Park, Eunice; Tran, Katherine; Miller, Jeffrey H.

    2010-01-01

    We have defined a sensitivity profile for 22 antibiotics by extending previous work testing the entire KEIO collection of close to 4,000 single-gene knockouts in Escherichia coli for increased susceptibility to 1 of 14 different antibiotics (ciprofloxacin, rifampin [rifampicin], vancomycin, ampicillin, sulfamethoxazole, gentamicin, metronidazole, streptomycin, fusidic acid, tetracycline, chloramphenicol, nitrofurantoin, erythromycin, and triclosan). We screened one or more subinhibitory concentrations of each antibiotic, generating more than 80,000 data points and allowing a reduction of the entire collection to a set of 283 strains that display significantly increased sensitivity to at least one of the antibiotics. We used this reduced set of strains to determine a profile for eight additional antibiotics (spectinomycin, cephradine, aztreonem, colistin, neomycin, enoxacin, tobramycin, and cefoxitin). The profiles for the 22 antibiotics represent a growing catalog of sensitivity fingerprints that can be separated into two components, multidrug-resistant mutants and those mutants that confer relatively specific sensitivity to the antibiotic or type of antibiotic tested. The latter group can be represented by a set of 20 to 60 strains that can be used for the rapid typing of antibiotics by generating a virtual bar code readout of the specific sensitivities. Taken together, these data reveal the complexity of intrinsic resistance and provide additional targets for the design of codrugs (or combinations of drugs) that potentiate existing antibiotics. PMID:20065048

  4. 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

  5. Recent results from APEX

    International Nuclear Information System (INIS)

    The focus of the APEX project is the study of the emission of electrons and positrons from collisions of heavy ions. However, scattering of this type involves intense Coulomb excitation of nuclear states so nuclear spectroscopy can never be far from one's mind. The authors also have close connections with the conference title; the detector which they describe does contain two large gamma arrays, each subtending 58% of 4π and containing 24 position sensitive detectors. Finally, they will see their research may have uncovered some interesting new spectroscopy in the actinides, so the connections between APEX and this conference may be closer than is first apparent

  6. 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

  7. 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....

  8. 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...

  9. [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

  10. 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...

  11. 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...

  12. Effect of Cyp27A1 gene dosage on atherosclerosis development in ApoE-knockout mice

    OpenAIRE

    Zurkinden, Line; Solcà, Curzio; Vögeli, Isabelle A.; Vogt, Bruno; Ackermann, Daniel; Erickson, Sandra K.; Frey, Felix J; Sviridov, Dmitri; Escher, Geneviève

    2014-01-01

    In humans, sterol 27-hydroxylase (CYP27A1) deficiency leads to cholesterol deposition in tendons and vasculature. Thus, in addition to its role in bile acid synthesis, where it converts cholesterol to 27-hydroxycholesterol (27-OHC), CYP27A1 may also be atheroprotective. Cyp27A1-deficient (Cyp27A1−/−) mice were crossed with apolipoprotein E (apoE)-deficient mice. Cyp27A1+/+/apoE−/− [ApoE-knockout (KO)], Cyp27A1+/−/apoE−/− heterozygous (het), and Cyp27A1−/−/apoE−/− [double-knockout (DKO)] mice ...

  13. Multiple phenotypic changes in mice after knockout of the B3gnt5 gene, encoding Lc3 synthase--a key enzyme in lacto-neolacto ganglioside synthesis

    Directory of Open Access Journals (Sweden)

    McLendon Roger E

    2010-11-01

    Full Text Available Abstract Background Ganglioside biosynthesis occurs through a multi-enzymatic pathway which at the lactosylceramide step is branched into several biosynthetic series. Lc3 synthase utilizes a variety of galactose-terminated glycolipids as acceptors by establishing a glycosidic bond in the beta-1,3-linkage to GlcNaAc to extend the lacto- and neolacto-series gangliosides. In order to examine the lacto-series ganglioside functions in mice, we used gene knockout technology to generate Lc3 synthase gene B3gnt5-deficient mice by two different strategies and compared the phenotypes of the two null mouse groups with each other and with their wild-type counterparts. Results B3gnt5 gene knockout mutant mice appeared normal in the embryonic stage and, if they survived delivery, remained normal during early life. However, about 9% developed early-stage growth retardation, 11% died postnatally in less than 2 months, and adults tended to die in 5-15 months, demonstrating splenomegaly and notably enlarged lymph nodes. Without lacto-neolacto series gangliosides, both homozygous and heterozygous mice gradually displayed fur loss or obesity, and breeding mice demonstrated reproductive defects. Furthermore, B3gnt5 gene knockout disrupted the functional integrity of B cells, as manifested by a decrease in B-cell numbers in the spleen, germinal center disappearance, and less efficiency to proliferate in hybridoma fusion. Conclusions These novel results demonstrate unequivocally that lacto-neolacto series gangliosides are essential to multiple physiological functions, especially the control of reproductive output, and spleen B-cell abnormality. We also report the generation of anti-IgG response against the lacto-series gangliosides 3'-isoLM1 and 3',6'-isoLD1.

  14. ALMA, APEX and beyond

    Science.gov (United States)

    Zwaan, M.; Testi, L.

    The Atacama Large Millimeter/submillimeter Array (ALMA) is currently being constructed at the 5000m Chajnantor plateau in the Chilean Andes. ALMA has been designed and is being built to deliver transformational science in the millimeter and submillimeter regime for many years to come. We briefly describe the project status and timeline. The Atacama Pathfinder Experiment (APEX), built at the same site, is already operational and proves to be an effective survey instrument. We discuss which niches in millimeter/submillimeter astronomy will remain open for a possible facility in Antarctica.

  15. 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

  16. Knockout of REST/NRSF shows that the protein is a potent repressor of neuronally expressed genes in non-neural tissues.

    Science.gov (United States)

    Jones, F S; Meech, R

    1999-05-01

    The protein repressor element 1 silencing transcription factor/neuron restrictive silencer factor (REST/NRSF) is a negative regulator of neuronal genes that contain a particular DNA sequence, the neuron restrictive silencer element (NRSE). REST is expressed ubiquitously in non-neural tissues but is down-regulated in neural precursors and turned off in postmitotic neurons, suggesting that it can act both to prevent extraneural expression of certain genes and to delay the differentiation of neuronal subtypes. In a recent paper, Chen et al.(1) describe the production of a null mutant for REST in mice and the mosaic inactivation of REST function in chicken embryos. Knockout of REST led to malformations in several non-neural tissues, as well as apoptosis and embryonic lethality in mice. In addition, the expression of several REST target genes was derepressed in non-neural tissues and in neural progenitors in both mouse and chicken embryos. These studies clearly demonstrate that active repression of tissue-specific genes is required for proper tissue differentiation during embryonic development. PMID:10376008

  17. 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 ...

  18. Recombinant adeno-associated virus type 2-mediated gene delivery into the Rpe65-/- knockout mouse eye results in limited rescue

    Science.gov (United States)

    Lai, Chooi-May; Yu, Meaghan JT; Brankov, Meliha; Barnett, Nigel L; Zhou, Xiaohuai; Redmond, T Michael; Narfstrom, Kristina; Rakoczy, P Elizabeth

    2004-01-01

    Background Leber's congenital amaurosis (LCA) is a severe form of retinal dystrophy. Mutations in the RPE65 gene, which is abundantly expressed in retinal pigment epithelial (RPE) cells, account for approximately 10–15% of LCA cases. In this study we used the high turnover, and rapid breeding and maturation time of the Rpe65-/- knockout mice to assess the efficacy of using rAAV-mediated gene therapy to replace the disrupted RPE65 gene. The potential for rAAV-mediated gene treatment of LCA was then analyzed by determining the pattern of RPE65 expression, the physiological and histological effects that it produced, and any improvement in visual function. Methods rAAV.RPE65 was injected into the subretinal space of Rpe65-/- knockout mice and control mice. Histological and immunohistological analyses were performed to evaluate any rescue of photoreceptors and to determine longevity and pattern of transgene expression. Electron microscopy was used to examine ultrastructural changes, and electroretinography was used to measure changes in visual function following rAAV.RPE65 injection. Results rAAV-mediated RPE65 expression was detected for up to 18 months post injection. The delivery of rAAV.RPE65 to Rpe65-/- mouse retinas resulted in a transient improvement in the maximum b-wave amplitude under both scotopic and photopic conditions (76% and 59% increase above uninjected controls, respectively) but no changes were observed in a-wave amplitude. However, this increase in b-wave amplitude was not accompanied by any slow down in photoreceptor degeneration or apoptotic cell death. Delivery of rAAV.RPE65 also resulted in a decrease in retinyl ester lipid droplets and an increase in short wavelength cone opsin-positive cells, suggesting that the recovery of RPE65 expression has long-term benefits for retinal health. Conclusion This work demonstrated the potential benefits of using the Rpe65-/- mice to study the effects and mechanism of rAAV.RPE65-mediated gene delivery into

  19. 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.

  20. 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.

  1. 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...

  2. 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

  3. 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.

  4. 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...

  5. Genetic transformation of the tomato pathogen Pyrenochaeta lycopersici allowed gene knockout using a split-marker approach.

    Science.gov (United States)

    Aragona, Maria; Valente, Maria Teresa

    2015-05-01

    Pyrenochaeta lycopersici, as other soil-transmitted fungal pathogens, generally received little attention compared to the pathogens affecting the aerial parts of the plants, although causing stunt and important fruit yield reduction of agronomic relevant crops. The scope of this study was to develop a system allowing to investigate the functional role of P. lycopersici genes putatively involved in the corky root rot of tomato. A genetic transformation system based on a split-marker approach was developed and tested to knock out a P. lycopersici gene encoding for a lytic polysaccharide monooxygenase (Plegl1) induced during the disease development. The regions flanking Plegl1 gene were fused with the overlapping parts of hygromycin marker gene, to favour homologous recombination. We were able to obtain four mutants not expressing the Plegl1 gene though, when tested on a susceptible tomato cultivar, Plegl1 mutants showed unaltered virulence, compared with the wild-type strain. The strategy illustrated in the present work demonstrated for the first time that homologous recombination occurs in P. lycopersici. Moreover, a transformation system mediated by Agrobacterium tumefaciens was established and stable genetic transformants have been obtained. The transformation systems developed represent important tools for investigating both the role of genes putatively involved in P. lycopersici interaction with host plant and the function of other physiological traits which emerged to be genetically expanded from the recent genome sequencing of this fungus. PMID:25413737

  6. Metabolic regulation of Escherichia coli and its phoB and phoR genes knockout mutants under phosphate and nitrogen limitations as well as at acidic condition

    Directory of Open Access Journals (Sweden)

    Shimizu Kazuyuki

    2011-05-01

    B regulated genes were down-regulated, while phoR and phoU changed little. The phoR gene knockout caused phoB gene to be down-regulated as well as PhoB regulated genes, while phoU and phoM changed little. The effect of pH together with lower P concentration on the metabolic regulation was also investigated. In accordance with up-regulation of arcA gene expression, the expressions of the TCA cycle genes such as sdhC and mdh were down-regulated at acidic condition. The gene expression of rpoS was up-regulated, and the expression of gadA was up-regulated at pH 6.0. In accordance with this, PhoB regulated genes were up-regulated in the wild type under P-rich and P-limited conditions at pH 6.0 as compared to those at pH 7.0. Moreover, the effect of nitrogen limitation on the metabolic regulation was investigated, where the result indicates that phoB gene was up-regulated, and PhoB regulated genes were also up-regulated under N-limitation, as well as nitrogen-regulated genes. Conclusion The present result shows the complicated nature of the metabolic regulation for the fermentation characteristics upon phosphate limitation, acidic condition, and nitrogen limitation based on the transcript levels of selected genes. The result implies that the regulations under phosphate limitation, acidic condition, and nitrogen limitation, which occur typically at the late growth phase of the batch culture, are interconnected through RpoS and RpoD together with Pho genes.

  7. 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

  8. 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

  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. 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...

  11. Effect of hyperlipidemia on the expression of circadian genes in apolipoprotein E knock-out atherosclerotic mice

    Directory of Open Access Journals (Sweden)

    Chen Sifeng

    2009-12-01

    Full Text Available Abstract Background Circadian patterns of cardiovascular vulnerability were well characterized, with a peak incidence of acute myocardial infarction and stroke secondary to atherosclerosis in the morning, which showed the circadian clock may take part in the pathological process of atherosclerosis induced by hyperlipidemia. Hence, the effect of hyperlipidemia on the expression of circadian genes was investigated in atherosclerotic mouse model. Results In apoE-/-mice on regular chow or high-fat diet, an atherosclerotic mouse model induced by heperlipidemia, we found that the peak concentration of serum lipids was showed four or eight hours later in apoE-/- mice, compared to C57BL/6J mice. During the artificial light period, a reduce in circulating level of serum lipids corresponded with the observed increase of the expression levels of some the transcription factors involved in lipid metabolism, such as PPARα and RXRα. Meanwhile, the expression of circadian genes was changed following with amplitude reduced or the peak mRNA level delayed. Conclusions Our studies indicated that heperlipidemia altered both the rhythmicity and expression of circadian genes. Diet-induced circadian disruption may affect the process of atherosclerosis and some acute cardiovascular disease.

  12. 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

  13. 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.

  14. 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)

    WU Xiaofeng; CAO Cuiping; XU Yaxiang; LU Xingmeng

    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.

  15. 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

  16. 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.

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

    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

  18. 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.

  19. 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

  20. 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.

  1. 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

  2. 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.

  3. 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.)

  4. 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...

  5. AP endonuclease knockdown enhances methyl methanesulfonate hypersensitivity of DNA polymerase β knockout mouse embryonic fibroblasts

    International Nuclear Information System (INIS)

    Apurinic/apyrimidinic (AP) endonuclease (Apex) is required for base excision repair (BER), which is the major mechanism of repair for small DNA lesions such as alkylated bases. Apex incises the DNA strand at an AP site to leave 3'-OH and 5'-deoxyribose phosphate (5'-dRp) termini. DNA polymerase β (PolB) plays a dominant role in single nucleotide (Sn-) BER by incorporating a nucleotide and removing 5'-dRp. Methyl methanesulfonate (MMS)-induced damage is repaired by Sn-BER, and thus mouse embryonic fibroblasts (MEFs) deficient in PolB show significantly increased sensitivity to MMS. However, the survival curve for PolB-knockout MEFs (PolBKOs) has a shoulder, and increased sensitivity is only apparent at relatively high MMS concentrations. In this study, we prepared Apex-knockdown/PolB-knockout MEFs (AKDBKOs) to examine whether BER is related to the apparent resistance of PolBKOs at low MMS concentrations. The viability of PolBKOs immediately after MMS treatment was significantly lower than that of wild-type MEFs, but there was essentially no effect of Apex-knockdown on cell viability in the presence or absence of PolB. In contrast, relative counts of MEFs after repair were decreased by Apex knockdown. Parental PolBKOs showed especially high sensitivity at >1.5 mM MMS, suggesting that PolBKOs have another repair mechanism in addition to PolB-dependent Sn-BER, and that the back-up mechanism is unable to repair damage induced by high MMS concentrations. Interestingly, AKDBKOs were hypersensitive to MMS in a relative cell growth assay, suggesting that MMS-induced damage in PolB-knockout MEFs is repaired by Apex-dependent repair mechanisms, presumably including long-patch BER. (author)

  6. 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.

  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

    OpenAIRE

    Sisay, S.; Pryce, G.; Jackson, S. J.; Tanner, C.; Ross, R A; Michael, G. J.; Selwood, D. L.; Giovannoni, G; Baker, D.

    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 treatmen...

  8. [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

  9. 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

  10. 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...

  11. 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...

  12. 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

  13. 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.)

  14. 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.)

  15. Orbital apex syndrome in a child

    OpenAIRE

    Sasindran, Vivek; Ravikumar, A.; Senthil

    2008-01-01

    A 8-year-old male presented with visual loss, diplopia, ptosis, pain behind the left eye, facial numbness and vomiting of one week duration. The ophthalmological, neurological and radiological examination showed a lesion of the left orbital apex with extension into the cavernous sinus. Examination of the nose and paranasal sinuses did not reveal any abnormality. Transnasal Endoscopic orbital decompression was performed and inflamed granulation tissue found in the orbital apex was removed. Mic...

  16. The Large APEX Bolometer Camera LABOCA

    OpenAIRE

    Siringo, G.; Kreysa, E.; Kovacs, 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.

    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.

  17. Knock-out of SO1377 gene, which encodes the member of a conserved hypothetical bacterial protein family COG2268, results in alteration of iron metabolism, increased spontaneous mutation and hydrogen peroxide sensitivity in Shewanella oneidensis MR-1

    Directory of Open Access Journals (Sweden)

    Klingeman Dawn M

    2006-04-01

    Full Text Available Abstract Background Shewanella oneidensis MR-1 is a facultative, gram-negative bacterium capable of coupling the oxidation of organic carbon to a wide range of electron acceptors such as oxygen, nitrate and metals, and has potential for bioremediation of heavy metal contaminated sites. The complete 5-Mb genome of S. oneidensis MR-1 was sequenced and standard sequence-comparison methods revealed approximately 42% of the MR-1 genome encodes proteins of unknown function. Defining the functions of hypothetical proteins is a great challenge and may need a systems approach. In this study, by using integrated approaches including whole genomic microarray and proteomics, we examined knockout effects of the gene encoding SO1377 (gi24372955, a member of the conserved, hypothetical, bacterial protein family COG2268 (Clusters of Orthologous Group in bacterium Shewanella oneidensis MR-1, under various physiological conditions. Results Compared with the wild-type strain, growth assays showed that the deletion mutant had a decreased growth rate when cultured aerobically, but not affected under anaerobic conditions. Whole-genome expression (RNA and protein profiles revealed numerous gene and protein expression changes relative to the wild-type control, including some involved in iron metabolism, oxidative damage protection and respiratory electron transfer, e. g. complex IV of the respiration chain. Although total intracellular iron levels remained unchanged, whole-cell electron paramagnetic resonance (EPR demonstrated that the level of free iron in mutant cells was 3 times less than that of the wild-type strain. Siderophore excretion in the mutant also decreased in iron-depleted medium. The mutant was more sensitive to hydrogen peroxide and gave rise to 100 times more colonies resistant to gentamicin or kanamycin. Conclusion Our results showed that the knock-out of SO1377 gene had pleiotropic effects and suggested that SO1377 may play a role in iron

  18. 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.

  19. 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.

  20. 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.

  1. 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.

  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

    Traumatic injury to the brain is one of the leading causes of injury-related death or disability, especially among young people. Inflammatory processes and oxidative stress likely underlie much of the damage elicited by injury, but the full repertoire of responses involved is not well known. 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...... 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. 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...

  4. Knockout beyond the dripline

    Energy Technology Data Exchange (ETDEWEB)

    Bonaccorso, A. [INFN, Sez. di Pisa, Largo B. Pontecorvo 3, 56127 Pisa (Italy); Charity, R. J. [Department of Chemistry, Washington University, St. Louis, Missouri 63130 (United States); Kumar, R. [Department of Physics, Deenbandhu Chhoturam University of Science and Technology, Murthal, Sonepat-131039 Haryana (India); Salvioni, G. [INFN, Sez. di Pisa, Largo B. Pontecorvo 3, 56127 Pisa, Italy and Dipartimento di Fisica, Università di Pisa, Largo B. Pontecorvo 3, 56127, Pisa (Italy)

    2015-02-24

    In this contribution, we will describe neutron and proton removal from {sup 9}C and {sup 7}Be which are two particularly interesting nuclei entering the nucleo-synthesis pp-chain [1, 2]. Neutron and proton removal reactions have been used in the past twenty years to probe the single-particle structure of exotic nuclei. The core parallel-momentum distribution can give information on the angular momentum and spin of the nucleon initial state while the total removal cross section is sensitive to the asymptotic part of the initial wave function and also to the reaction mechanism. Because knockout is a peripheral reaction from which the Asymptotic Normalization Constant (ANC) of the single-particle wave function can be extracted, it has been used as an indirect method to obtain the rate of reactions like {sup 8}B(p,γ){sup 9}C or {sup 7}Be(p,γ){sup 8}B. Nucleon removal has recently been applied by the HiRA collaboration [3] to situations in which the remaining “core” is beyond the drip line, such as {sup 8}C and {sup 6}Be, unbound by one or more protons, and whose excitation-energy spectrum can be obtained by the invariant-mass method. By gating on the ground-state peak, “core” parallel-momentum distributions and total knockout cross sections have been obtained similar to previous studies with well-bound “cores”. In addition for each projectile, knock out to final bound states has also been obtained in several cases. We will report on the theoretical description and comparison to this experimental data for a few cases for which advances in the accuracy of the transfer-to-the continuum model [4, 5] have been made [6]. These include the use, when available, of “ab-initio” overlaps for the initial state [7] and in particular their ANC values [8]. Also, the construction of a nucleus-target folding potential for the treatment of the core-target S-matrix [9] using for the cores “ab-initio” densities [10] and state-of-the-art n−{sup 9}Be optical

  5. 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

  6. [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

  7. 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.

  8. 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

  9. 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.

  10. Directed evolution of APEX2 for electron microscopy and proteomics

    OpenAIRE

    Lam, Stephanie S.; Martell, Jeffrey D.; Kamer, Kimberli J; Deerinck, Thomas J.; Ellisman, Mark H.; Mootha, Vamsi K.; Ting, Alice Y.

    2014-01-01

    APEX is an engineered peroxidase that functions both as an electron microscopy tag, and as a promiscuous labeling enzyme for live-cell proteomics. Because the limited sensitivity of APEX precludes applications requiring low APEX expression, we used yeast display evolution to improve its catalytic efficiency. Our evolved APEX2 is far more active in cells, enabling the superior enrichment of endogenous mitochondrial and endoplasmic reticulum membrane proteins and the use of electron microscopy ...

  11. 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

  12. 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

  13. 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...

  14. 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.

  15. 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.

  16. 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.

  17. 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.

  18. 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.

  19. 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.

  20. Generation of Fgfr3 Conditional Knockout Mice

    Directory of Open Access Journals (Sweden)

    Nan Su, Xiaoling Xu, Cuiling Li, Qifen He, Ling Zhao, Can Li, Siyu Chen, Fengtao Luo, Lingxian Yi, Xiaolan Du, Haiyang Huang, Chuxia Deng, Lin Chen

    2010-01-01

    Full Text Available Fibroblast growth factor receptor 3 (FGFR3, highly conserved in both humans and murine, is one of key tyrosine kinase receptors for FGF. FGFR3 is expressed in different tissues, including cartilage, brain, kidney, and intestine at different development stages. Conventional knockout of Fgfr3 alleles leads to short life span, and overgrowth of bone. In clinic, human FGFR3 mutations are responsible for three different types of chondrodysplasia syndromes including achondroplasia (ACH, hypochondroplasia (HCH and thanatophoric dysplasia (TD. For better understanding of the roles of FGFR3 in different tissues at different stages of development and in pathological conditions, we generated Fgfr3 conditional knockout mice in which loxp sites flank exons 9-10 in the Fgfr3 allele. We also demonstrated that Cre-mediated recombination using Col2a1-Cre, a Cre line expressed in chondrocyte during bone development, results in specific deletion of the gene in tissues containing cartilage. This animal model will be useful to study distinct roles of FGFR3 in different tissues at different ages.

  1. Cocaine conditioned locomotion in dopamine transporter, norepinephrine transporter and serotonin transporter knockout mice

    OpenAIRE

    Hall, F. Scott; Li, Xiao-Fei; Randall-Thompson, Jovita; Sora, Ichiro; Murphy, Dennis L.; Lesch, Klaus-Peter; Caron, Marc; Uhl, George R

    2009-01-01

    The behavioral effects of cocaine are affected by gene knockout of the dopamine transporter (DAT), the serotonin transporter (SERT) and the norepinephrine transporter (NET). The relative involvement of each of these transporters varies depending on the particular behavioral response to cocaine considered, as well as on other factors such as genetic background of the subjects. Interestingly, the effects of these gene knockouts on cocaine induced locomotion are quite different from those on rew...

  2. Transgenic mice: beyond the knockout

    OpenAIRE

    Miller, R. Lance

    2010-01-01

    Transgenic mice have had a tremendous impact on biomedical research. Most researchers are familiar with transgenic mice that carry Cre recombinase (Cre) and how they are used to create conditional knockouts. However, some researchers are less familiar with many of the other types of transgenic mice and their applications. For example, transgenic mice can be used to study biochemical and molecular pathways in primary cultures and cell suspensions derived from transgenic mice, cell-cell interac...

  3. Argonne Plasma Engineering Experiment (APEX) Tokamak

    International Nuclear Information System (INIS)

    The Argonne Plasma Engineering Experiment (APEX) Tokamak was designed to provide hot plasmas for reactor-relevant experiments with rf heating (current drive) and plasma wall experiments, principally in-situ low-Z wall coating and maintenance. The device, sized to produce energetic plasmas at minimum cost, is small (R = 51 cm, r = 15 cm) but capable of high currents (100 kA) and long pulse durations (100 ms). A design using an iron central core with no return legs, pure tension tapewound toroidal field coils, digital radial position control, and UHV vacuum technology was used. Diagnostics include monochrometers, x-ray detectors, and a microwave interferometer and radiometer for density and temperature measurements. Stable 100 ms shots were produced with electron temperatures in the range 500 to 1000 eV. Initial results included studies of thermal desorption and recoating of wall materials

  4. 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.

  5. 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.

  6. 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.

  7. 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

  8. Reaching for the APEX at Ames

    Science.gov (United States)

    Kohut, Matthew

    2008-01-01

    The multidimensional design of the APEX program is the result of an extensive research and development effort dating back nearly a decade. "In the late 1990s and early 2000, we were pretty successful at getting new research and technology projects here at the center," Johnson says, "and we had a lack of critical mass of project managers. We were taking people who were primarily researchers and putting them in the position of managing projects." Smith and Johnson held a series of workshops across the center during 2000 and 2001 to gather feedback about how to address this issue. When they briefed the center's senior management on their findings, one of the top recommendations was to establish a project manager development program at Ames. At that point, they cast a wide net for ideas and information. "We did centerwide needs assessment, we did focus groups, we did surveys," Smith says. "We came up with a proposal for what a program would look like, tying in what we knew about the Academy of Program1 Project Leadership (now the Academy for Program/Project and Engineering Leadership, or APPEL), what we've seen at other centers, what other centers have tried. We were always checking to make sure our program mapped to APPEL. We also looked at the PMI [Project Management Institute] model, INCOSE [International Council on Systems Engineering], CMMI [Capability Maturity Model Integration], you name it." "We had a lot of conversations with the Jet Propulsion Lab and Goddard," Johnson adds. "We saw those centers as models for what Ames was aspiring to be in terms of a center for managing space flight missions." Their research confirmed what they already knew-that strong practitioner involvement would be critical to their program design process. 'XPEX is for the practitioner by the practitioner," Smith says. "They have to be a part of designing it. Otherwise there's no way we could design a program that meets their needs." At the same time that they worked at the grassroots

  9. Reduced cocaine-seeking behavior in heterozygous BDNF knockout rats

    OpenAIRE

    St. Laurent, Robyn; Helm, Samuel R.; Glenn, Melissa J.

    2013-01-01

    Cocaine generates drug-seeking behavior by creating long-lasting changes in the reward pathway. The role of the growth factor, brain-derived neurotrophic factor (BDNF) in facilitating these changes was investigated in the present report with a genetic rat model. Using conditioned place preference, the current study investigated the hypothesis that a partial knockout of the BDNF gene in rats (BDNF+/−) would attenuate the rewarding effects of cocaine. Wildtype rats exposed to cocaine exhibited ...

  10. Analysis of Microsatellite Polymorphism in Inbred Knockout Mice

    OpenAIRE

    Zuo, Baofen; Du, Xiaoyan; Zhao, Jing; Yang, Huixin; Wang, Chao; Wu, Yanhua; LU, JING; Wang, Ying; Chen, Zhenwen

    2012-01-01

    Previously, we found that the genotype of 42 out of 198 mouse microsatellite loci, which are distributed among all chromosomes except the Y chromosome, changed from monomorphism to polymorphism (CMP) in a genetically modified inbred mouse strain. In this study, we further examined whether CMP also relates to the homologous recombination in gene knockout (KO) mouse strains. The same 42 microsatellite loci were analyzed by polymerase chain reaction (PCR) in 29 KO inbred mouse strains via short ...

  11. 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.

  12. 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.

  13. Localization of the prostatic apex for radiotherapy treatment planning

    International Nuclear Information System (INIS)

    Purpose/Objective: There is no consensus on the optimal method for localizing the prostatic apex in patients with early stage adenocarcinoma of the prostate. Some radiation oncologists have recommended that transrectal ultrasound or MRI scans be used to define the inferior border of radiation portals. The purpose of this prospective study is to assess the ability of retrograde urethrograms and CT scans to accurately define the prostatic apex in the craniocaudad dimension, using urethroscopy as a reference. Materials and Methods: Following construction of an Alpha cradle, plain radiographs of the pelvis were obtained in 15 patients with early stage adenocarcinoma of the prostate, with the tip of a urethroscope placed at the superior border of the external sphincter (which most closely approximates the prostatic apex). The scope was then withdrawn, and a retrograde urethrogram was performed. Immediately afterwards, a treatment planning CT scan of the pelvis was obtained. Since differential filling of the bladder and rectum affects the position of the prostatic apex, patients voided prior to rather than in between the 3 consecutive studies. Results: The urethroscopy-defined prostatic apex was located 28 ± 3 mm (mean ± SE) superior to the ischial tuberosities, 12 ± 1 mm (mean ± SE) superior to the urethrogram tip and 8 ± 2 mm (mean ± SE) superior to the CT-defined apex. Placement of the inferior border of the radiation portals at the ischial tuberosities would have resulted in irradiation of > 20 mm membranous and spongy urethra in all of the patients. Conclusion: Retrograde urethrograms provide more helpful information than CT scans with regard to localization of the prostatic apex and are more cost effective than sonograms or MRI scans. The prostatic apex is typically 12 mm superior to the urethrogram tip with little variability. Retrograde urethrograms allow one to spare as much urethra as possible in the radiation portals, which should theoretically reduce

  14. 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...

  15. 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

  16. 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.

  17. Knockout confirmation for Hurries: rapid genotype identification of Trypanosoma cruzi transfectants by polymerase chain reaction directly from liquid culture

    OpenAIRE

    2014-01-01

    Gene knockout is a widely used approach to evaluate loss-of-function phenotypes and it can be facilitated by the incorporation of a DNA cassette having a drug-selectable marker. Confirmation of the correct knockout cassette insertion is an important step in gene removal validation and has generally been performed by polymerase chain reaction (PCR) assays following a time-consuming DNA extraction step. Here, we show a rapid procedure for the identification of Trypanosoma cruzi transfectants by...

  18. [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

  19. 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.

  20. 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.

  1. Orbital apex syndrome associated with herpes zoster ophthalmicus

    Directory of Open Access Journals (Sweden)

    Kurimoto T

    2011-11-01

    Full Text Available Takuji Kurimoto1, Masahiro Tonari1, Norihiko Ishizaki1, Mitsuhiro Monta2, Saori Hirata2, Hidehiro Oku1, Jun Sugasawa1, Tsunehiko Ikeda11Department of Ophthalmology, Osaka Medical College, 2Department of Ophthalmology, Shitennoji Hospital, Osaka, JapanAbstract: We report our findings for a patient with orbital apex syndrome associated with herpes zoster ophthalmicus. Our patient was initially admitted to a neighborhood hospital because of nausea and loss of appetite of 10 days' duration. The day after hospitalization, she developed skin vesicles along the first division of the trigeminal nerve, with severe lid swelling and conjunctival injection. On suspicion of meningoencephalitis caused by varicella zoster virus, antiviral therapy with vidarabine and betamethasone was started. Seventeen days later, complete ptosis and ophthalmoplegia developed in the right eye. The light reflex in the right eye was absent and anisocoria was present, with the right pupil larger than the left. Fat-suppressed enhanced T1-weighted magnetic resonance images showed high intensity areas in the muscle cone, cavernous sinus, and orbital optic nerve sheath. Our patient was diagnosed with orbital apex syndrome, and because of skin vesicles in the first division of the trigeminal nerve, the orbital apex syndrome was considered to be caused by herpes zoster ophthalmicus. After the patient was transferred to our hospital, prednisolone 60 mg and vidarabine antiviral therapy was started, and fever and headaches disappeared five days later. The ophthalmoplegia and optic neuritis, but not the anisocoria, gradually resolved during tapering of oral therapy. From the clinical findings and course, the cause of the orbital apex syndrome was most likely invasion of the orbital apex and cavernous sinus by the herpes virus through the trigeminal nerve ganglia.Keywords: varicella zoster virus, orbital apex syndrome, herpes zoster ophthalmicus, complete ophthalmoplegia

  2. 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

  3. 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.

  4. 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

  5. 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

  6. 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

  7. 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

  8. 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.

  9. 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

  10. Contributions of Selective Knockout Studies to Understanding Cholinesterase Disposition and Function

    OpenAIRE

    Camp, Shelley; ZHANG, LIMIN; Krejci, Eric; Dobbertin, Alexandre; Bernard, Véronique; Girard, Emmanuelle; Duysen, Ellen G.; Lockridge, Oksana; De Jaco, Antonella; Taylor, Palmer

    2010-01-01

    The complete knockout of the acetylcholinesterase gene (AChE) in the mouse yielded a surprising phenotype that could not have been predicted from deletion of the cholinesterase genes in Drosophila, that of a living, but functionally compromised animal. The phenotype of this animal showed a sufficient compromise in motor function that precluded precise characterization of central and peripheral nervous functional deficits. Since AChE in mammals is encoded by a single gene with alternative spli...

  11. 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…

  12. 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

  13. 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

  14. 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.

  15. 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.

  16. Petrous Apex Cholesteatoma: Exteriorization vs. Subtotal Petrosectomy with Obliteration

    OpenAIRE

    Pyle, G. Mark; Wiet, Richard J.

    1991-01-01

    True cholesteatomas developing medial to the labyrinth and facial nerve with extension into the petrous apex are uncommon lesions. More recent techniques of total matrix removal and obliteration require the otoneurosurgeon to be knowledgeable of intratemporal facial nerve mobilization techniques. Total cholesteatoma matrix removal and obliteration may obviate the need for frequent care of the exteriorized cavity. However, these procedures can be associated with extensive recurrence from resid...

  17. 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.

  18. 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.

  19. 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

  20. Possible use of calcifying nanoparticles in immature root apex treatment

    Directory of Open Access Journals (Sweden)

    Mohammed S Alenazy

    2014-01-01

    Full Text Available Introduction: There are reports in the literature, which describe different techniques and materials in the challenging management of thin dentin walls and immature root apex. It has been suggested that calcifying nanoparticles (CNPs could be used in the management of these conditions. The Hypothesis: Compositionally modified CNPs made into a paste could become efficacious in managing thin dentin walls and immature root apex. Calcium and phosphate ions when mixed with CNPs could form a synthetic nanopaste that clinicians could use to manage thin dentin walls and to get a biological seal for immature root apex. Evaluation of the Hypothesis: CNPs can replicate and could facilitate the aggregations of calcium hydroxyapatite to produce a self-surrounding shell. These characteristics of CNPs could be used through their biomineralization process as initial nidus of calcification for further calcification progression to achieve total biological apical seal. If the hypothesis could be supported by biomineralization behavior of the paste (CNPs, Ca2 + , and PO4− , a new therapeutic agent would have been added to the armamentarium of endodontists. There is need for more in vivo and in vitro investigations of modified nanopaste to manage these conditions.

  1. Impaired Extinction of Learned Contextual Fear Memory in Early Growth Response 1 Knockout Mice

    OpenAIRE

    Han, Seungrie; Hong, Soontaek; Mo, Jiwon; Lee, Dongmin; Choi, Eunju; Choi, June-Seek; Sun, Woong; Lee, Hyun Woo; Kim, Hyun

    2014-01-01

    Inductive expression of early growth response 1 (Egr-1) in neurons is associated with many forms of neuronal activity. However, only a few Egr-1 target genes are known in the brain. The results of this study demonstrate that Egr-1 knockout (KO) mice display impaired contextual extinction learning and normal fear acquisition relative to wild-type (WT) control animals. Genome-wide microarray experiments revealed 368 differentially expressed genes in the hippocampus of Egr-1 WT exposed to differ...

  2. A novel CYP17A1 deletion causes a functional knockout of the steroid enzyme 17-hydroxylase and 17,20-lyase in a Turkish family and illustrates the precise role of the CYP17A1 gene.

    Science.gov (United States)

    Camats, Núria; Üstyol, Ala; Atabek, Mehmet Emre; Dick, Bernhard; Flück, Christa E

    2015-10-01

    A novel homozygous long-range deletion of the CYP17A1 gene abolished protein expression and caused the severest form of 17-hydroxylase deficiency in one kindred of a Turkish family. The affected subjects presented with 46,XY sex reversal and 46,XX lack of pubertal development as well as severe hypertension. PMID:26509008

  3. Mup-knockout mice generated through CRISPR/Cas9-mediated deletion for use in urinary protein analysis.

    Science.gov (United States)

    Yang, Haixia; Zhang, Wei; Lu, Shan; Lu, Guangqing; Zhang, Hongjuan; Zhuang, Yinghua; Wang, Yue; Dong, Mengqiu; Zhang, Yu; Zhou, Xingang; Wang, Peng; Yu, Lei; Wang, Fengchao; Chen, Liang

    2016-05-01

    Major urinary proteins (MUPs) are the most abundant protein species in mouse urine, accounting for more than 90% of total protein content. Twenty-one Mup genes and 21 pseudogenes are clustered in a region of around 2 megabase pairs (Mbp) on chromosome 4. A Mup-knockout mouse model would greatly facilitate researches in the field of proteomic analysis of mouse urine. Here, we report the successful knockout of the Mup gene cluster of 2.2 Mbp using the CRISPR/Cas9 system. Homozygous Mup-knockout mice survived to adulthood and exhibited no obvious defects. The patterns of the proteomes of non-MUP urinary proteins in homozygous Mup-knockout mice were similar to those of wild-type mice judged by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The sensitivity of enzyme-linked immunosorbent assay to detect non-MUP urinary protein was significantly enhanced in Mup-knockout mice. In short, we have developed a Mup-knockout mouse model. This mouse model will be useful for the research of urinary biomarker testing that may have relevance for humans. PMID:26851484

  4. Knockout reactions in atomic and nuclear physics

    International Nuclear Information System (INIS)

    In a knockout experiment the momenta of a projectile before and after the collision and of a knocked-out particle are all measured, so that the recoil momentum of the residual system is known by subtraction. The atomic (e,2e) experiments are very much more accurate and detailed than present nuclear experiments. The (e,2e) reaction on argon is used to illustrate the principles involved. Other experiments involve the (p,2p) and (e,e'p) reactions

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

    International Nuclear Information System (INIS)

    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

  6. 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.

  7. 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.

  8. APEX CO(3-2) observations of NGC6822

    OpenAIRE

    De Rijcke, S.; Buyle, P.; Cannon, J.; Walter, F.; Lundgren, A.; Michielsen, D.; Dejonghe, H.

    2006-01-01

    We observed the CO(3-2) emission of the emission-line regions HubbleI, HubbleV, HubbleX, Holmberg 18, and the stellar emission-line object S28 in NGC6822 with the ESO Atacama Pathfinder Experiment (APEX) 12m telescope as part of its science verification. The very low system temperature of 130-180K enabled us to achieve detections in 4 single pointings and in a high spatial resolution 70''x70'' map of HubbleV. We compare the spectra with HI observations, obtained with the Australia Telescope C...

  9. 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.

  10. APEX accelerator cycle for transmutation of long-lived fission wastes

    International Nuclear Information System (INIS)

    Based on preliminary studies, some conclusions can be drawn concerning the Accelerator Fuel Enricher and Fission Product Exterminator (APEX). APEX-1 and APEX-2 systems can destroy TU's, 137Cs, and 90Sr at acceptable cost and efficiency. The principal difference between APEX-1 and APEX-2 is the in-reactor and in-circuit inventory of 137Cs and 90Sr. Stable and low hazard wastes can be disposed of by burial. Accelerator breeders can effectively sustain a fission reactor economy indefinitely. Military waste can be blended into commercial fuel cycle for transmutation. Accelerator and target technologies appear practical and could be developed in a few years. More detailed studies are needed to better define the technical and economic features of the LAFER and APEX cycles, so that comparative assessments can be made between these cycles, as well as with other transmutation and waste disposal concepts

  11. 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

  12. 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.

  13. Hypercarnivorous apex predator could provide ecosystem services by dispersing seeds

    Science.gov (United States)

    Sarasola, José Hernán; Zanón-Martínez, Juan Ignacio; Costán, Andrea Silvina; Ripple, William J.

    2016-01-01

    Large “hypercarnivorous” felids are recognized for their role as apex predators and hence as key elements in food webs and ecosystem functioning through competition and depredation. Here we show that cougars (Puma concolor), one of the largest and the most widely ranging apex felid predators with a strictly carnivorous diet, could also be effective secondary long distance seed dispersers, potentially establishing direct and non-herbivore mediated interactions with plant species at the bottom of the food web. Cougars accidently ingest and disseminate large amounts of seeds (31,678 seeds in 123 scats) of plant species initially consumed by their main prey, the Eared Dove Zenaida auriculata. The germination potential of seeds for the three plant species most abundantly found in cougar scats (19,570 seeds) was not significantly different from that observed in seeds obtained from dove gizzards, indicating that seed passage through cougar guts did not affect seed germination. Considering the estimated cougar density in our study area, dispersal of seeds by cougars could allow a mean, annual seed spread of ~5,000 seeds per km2. Our results demonstrate that strictly carnivorous, felid predators could have broad and overlooked ecological functions related to ecosystem structuring and functioning. PMID:26791932

  14. 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

  15. 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.

  16. 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.

  17. Systematic construction of a conceptual minimal model of plasma cholesterol levels based on knockout mouse phenotypes.

    Science.gov (United States)

    van de Pas, Niek C A; Soffers, Ans E M F; Freidig, Andreas P; van Ommen, Ben; Woutersen, Ruud A; Rietjens, Ivonne M C M; de Graaf, Albert A

    2010-06-01

    Elevated plasma cholesterol, a well-known risk factor for cardiovascular diseases, is the result of the activity of many genes and their encoded proteins in a complex physiological network. We aim to develop a minimal kinetic computational model for predicting plasma cholesterol levels. To define the scope of this model, it is essential to discriminate between important and less important processes influencing plasma cholesterol levels. To this end, we performed a systematic review of mouse knockout strains and used the resulting dataset, named KOMDIP, for the identification of key genes that determine plasma cholesterol levels. Based on the described phenotype of mouse knockout models, 36 of the 120 evaluated genes were marked as key genes that have a pronounced effect on the plasma cholesterol concentration. The key genes include well-known genes, e.g., Apoe and Ldlr, as well as genes hardly linked to cholesterol metabolism so far, e.g., Plagl2 and Slc37a4. Based on the catalytic function of the genes, a minimal conceptual model was defined. A comparison with nine conceptual models from literature revealed that each of the individual published models is less complete than our model. Concluding, we have developed a conceptual model that can be used to develop a physiologically based kinetic model to quantitatively predict plasma cholesterol levels. PMID:20176131

  18. 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.

  19. 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

  20. 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...

  1. Comprehensive behavioral analysis of pituitary adenylate cyclase-activating polypeptide (PACAP) knockout mice

    OpenAIRE

    Koichi Tanda; Norihito Shintani; Akemichi Baba; Hitoshi Hashimoto; Tsuyoshi Miyakawa

    2012-01-01

    Pituitary adenylate cyclase-activating polypeptide (PACAP) is a neuropeptide acting as a neurotransmitter, neuromodulator, or neurotrophic factor. PACAP is widely expressed throughout the brain and exerts its functions through the PACAP-specific receptor (PAC1). Recent studies reveal that genetic variants of the PACAP and PAC1 genes are associated with mental disorders, and several behavioral abnormalities of PACAP knockout (KO) mice are reported. However, an insufficient number of backcrosse...

  2. 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...

  3. 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.

  4. 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...

  5. Reduced cortical BDNF expression and aberrant memory in Carf knockout mice

    OpenAIRE

    McDowell, Kelli A.; Hutchinson, Ashley N.; Wong-Goodrich, Sarah J.E.; Presby, Matthew M.; Su, Dan; Rodriguiz, Ramona M.; Law, Krystal C.; Williams, Christina L.; Wetsel, William C.; West, Anne E.

    2010-01-01

    Transcription factors are a key point of convergence between the cell-intrinsic and extracellular signals that guide synaptic development and brain plasticity. Calcium-Response Factor (CaRF) is a unique transcription factor first identified as a binding protein for a calcium-response element in the gene encoding Brain-Derived Neurotrophic Factor (Bdnf). We have now generated Carf knockout (KO) mice to characterize the function of this factor in vivo. Intriguingly, Carf KO mice have selectivel...

  6. Efficient conditional knockout targeting vector construction using co-selection BAC recombineering (CoSBR)

    OpenAIRE

    Robert J. Newman; Roose-Girma, Merone; Warming, Søren

    2015-01-01

    A simple and efficient strategy for Bacterial Artificial Chromosome (BAC) recombineering based on co-selection is described. We show that it is possible to efficiently modify two positions of a BAC simultaneously by co-transformation of a single-stranded DNA oligo and a double-stranded selection cassette. The use of co-selection BAC recombineering reduces the DNA manipulation needed to make a conditional knockout gene targeting vector to only two steps: a single round of BAC modification foll...

  7. 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.

  8. 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

  9. 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

    International Nuclear Information System (INIS)

    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

  10. 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

  11. 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.

  12. 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

  13. 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

  14. Treatment of traumatized teeth with incomplete apex – apexification

    Directory of Open Access Journals (Sweden)

    Melissa Andréia MARCHESAN

    2008-04-01

    Full Text Available Introduction and objective: Two cases reports of traumatized teethwith incomplete apexes and no tooth fracture. Case report: In thesecases, instrumentation should be performed with no pressure to thewalls due to the decreased thickness, therefore, the irrigating solution is fundamental to achieve ideal biomechanical preparation. Calcium hydroxide intracanal dressings between sessions were applied at 7 days,15 days, 3 changes every 30 days and a change after 3 months, until apical barrier formation, and sealing of the root canal with the single-gutta-percha point technique with Grossman’s cement. Conclusion:The used clinical protocol efficiently formed the calcified root barrier and biological and functional equilibrium was achieved guaranteeing the treatment success.

  15. 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.

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

    DEFF Research Database (Denmark)

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

    2014-01-01

    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...... 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...... 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...

  17. Localization of the prostatic apex using CT for radiation treatment planning

    International Nuclear Information System (INIS)

    Objective: In this retrospective study, we analyzed the magnetic resonance imaging (MRI) and computed tomography (CT) scans of patients with prostate cancer to investigate the relationship between the apex of prostate and the anatomic structures visible in CT, and to provide evidence for localizing the prostatic apex in radiation treatment planning. Methods: MRI and CT scans from 108 patients with prostate cancer were analyzed to measure the distance between the prostatic apex and the bottom of ischial tuberosities, the bottom of obturator foramen, the bottom of pubic symphysis and the bulb of the penis. The volume of prostate was calculated and the relationship between the size of the prostate and the localization of the prostatic apex was analyzed. Results: The prostatic apex is located 13.1 mm ± 3.3 mm superior to the bulb of the penis, 11.0 mm ± 5.4 mm superior to the bottom of obturator foramen, 31.3 mm ± 5.5 mm superior to the bottom of ischial tuberosities, and 7.1 mm ± 4.7 mm superior to the bottom of obturator foramen. There was no correlation between the size of prostate and the localization of the prostatic apex (R =0.07, -0.33, all P > 0.05). Conclusions: Ninety-five percent of patients had a prostatic apex that is above the bulb of the penis 6 mm, and 100% of patients had a prostatic apex that is above the bottom of obturator foramen. (authors)

  18. Surgical Approaches to the Petrous Apex: Distances and Relations with Cranial Morphology

    OpenAIRE

    MURILO S. MENESES; Moreira, Ana Lucila; Kelly C. Bordignon; Ari A. Pedrozo; Ramina, Ricardo; Nikoski, Jeziel G.

    2004-01-01

    Different pathologies may be located at the petrous apex. The primary surgical approaches used to remove such lesions are the pterional, subtemporal, presigmoid, and retrosigmoid. Each has advantages and disadvantages, and the distance to the petrous apex varies with the approach. Anatomical variations in cranial morphology may interfere with these distances. Dolichocephalic skulls have a longer anteroposterior axis than brachycephalic ones.

  19. METRO-APEX Volume 15.1: Industrialist's Manual No. 5, Caesar's Rendering Plant. Revised.

    Science.gov (United States)

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

    The Industrialist's Manual No. 5 (Caesar's Rendering Plant) 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…

  20. 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

  1. 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

  2. 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.

  3. 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.

  4. 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...

  5. 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...

  6. 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...

  7. Generation of ERα-floxed and knockout mice using the Cre/LoxP system

    International Nuclear Information System (INIS)

    Highlights: ► ERα floxed and knockout mice were generated. ► Disruption of the ERα gene results in sterility in both male and female mice. ► ERα−/− mice have ovaries with hemorrhagic follicles and hypoplastic uterus. ► Female ERα−/− mice develop obesity. -- Abstract: Estrogen receptor alpha (ERα) 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 ERα mouse line that can be used to knock out ERα in selected tissues by using the Cre/LoxP system. In this study, we established a new ERα knockout mouse line by crossing the floxed ERα mice with Cre deleter mice. Here we show that genetic disruption of the ERα 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 ERα is necessary for reproductive tract development and has important functions as a regulator of metabolism in females.

  8. S100A9 Knockout Decreases the Memory Impairment and Neuropathology in Crossbreed Mice of Tg2576 and S100A9 Knockout Mice Model

    OpenAIRE

    Kim, Hee Jin; Chang, Keun-A; Ha, Tae-Young; Kim, Jeonga; Ha, Sungji; Shin, Ki-Young; Moon, Cheil; Nacken, Wolfgang; Kim, Hye-Sun; Suh, Yoo-Hun

    2014-01-01

    Our previous study presented evidence that the inflammation-related S100A9 gene is significantly upregulated in the brains of Alzheimer's disease (AD) animal models and human AD patients. In addition, experiments have shown that knockdown of S100A9 expression improves cognition function in AD model mice (Tg2576), and these animals exhibit reduced amyloid plaque burden. In this study, we established a new transgenic animal model of AD by crossbreeding the Tg2576 mouse with the S100A9 knockout ...

  9. Assessment of tumors of the lung apex by imaging techniques

    International Nuclear Information System (INIS)

    The purpose of this study was to analyze the value of MR in the preoperative staging of tumors of the lung apex and detection of local invasion of adjacent structures to determine its influence on the therapeutic approach. We obtained plain X-ray images in two planes, as well as CT and Mr images, in 12 patients with Pan coast tumor in whom there was surgical (n=8) or clinical (n=4) evidence of invasion. The objective was to assess local infiltration of brain stem and chest wall soft tissue, enveloping of the subclavian artery, substantial involvement of the brachial plexus and destruction of the vertebral body. In our series, MR was superior to the other imaging techniques in predicting the involvement of the structures surrounding the tumor. In conclusion, MR should be performed in a patient diagnosed by plain radiography as having an apical tumors to assess local tumor extension, while CT should be done to detect mediastinal lymph node involvement and distant metastases. 19 refs

  10. Acute orbital apex syndrome and rhino-orbito-cerebral mucormycosis

    Directory of Open Access Journals (Sweden)

    Anders UM

    2015-04-01

    Full Text Available Ursula M Anders,1 Elise J Taylor,1 Joseph R Martel,1–3 James B Martel1–3 1Research Center, Martel Eye Medical Group, Rancho Cordova, 2Graduate Medical Education, California Northstate University College of Medicine, Elk Grove, 3Department of Ophthalmology, Dignity Health, Carmichael, CA, USA Purpose: To demonstrate the successful clinical identification and management of rhino-orbital mucormycosis, a fungal infection with a high mortality rate. Patients and methods: A diabetic male patient with a headache and orbital apex syndrome in the right eye was examined using computed tomography (CT and magnetic resonance imaging (MRI for a possible fungal infection. Endoscopic surgical resection was performed and a pathology sample was taken. Specimens were prepared with Gömöri methenamine silver and hematoxylin and eosin staining. The patient was treated with liposomal amphotericin B 400 mg daily, followed by posaconazole 400 mg twice daily. Results: CT and MRI revealed a mass of the right sphenoid spreading into the orbit, indicative of a fungal infection. The biopsy confirmed the diagnosis of mucormycosis. Complete recovery of eyelid and oculomotor function was achieved after 10 months of treatment, although the patient continues to suffer from irreversible blindness in the right eye due to optic nerve atrophy. He has been without signs or symptoms of recurrence. Conclusion: Patients with rhino-orbito-cerebral mucormycosis need extensive surgical and medical treatment to maximize outcomes. Success requires multidisciplinary management. Keywords: ophthalmoplegia, sixth nerve palsy, diabetes mellitus, nephrotoxicity, amphotericin B, posaconazole

  11. Body condition predicts energy stores in apex predatory sharks.

    Science.gov (United States)

    Gallagher, Austin J; Wagner, Dominique N; Irschick, Duncan J; Hammerschlag, Neil

    2014-01-01

    Animal condition typically reflects the accumulation of energy stores (e.g. fatty acids), which can influence an individual's decision to undertake challenging life-history events, such as migration and reproduction. Accordingly, researchers often use measures of animal body size and/or weight as an index of condition. However, values of condition, such as fatty acid levels, may not always reflect the physiological state of animals accurately. While the relationships between condition indices and energy stores have been explored in some species (e.g. birds), they have yet to be examined in top predatory fishes, which often undertake extensive and energetically expensive migrations. We used an apex predatory shark (Galeocerdo cuvier, the tiger shark) as a model species to evaluate the relationship between triglycerides (energy metabolite) and a metric of overall body condition. We captured, blood sampled, measured and released 28 sharks (size range 125-303 cm pre-caudal length). In the laboratory, we assayed each plasma sample for triglyceride values. We detected a positive and significant relationship between condition and triglyceride values (P fisheries, because these individuals are likely to have the highest potential for successful reproduction. Our results suggest that researchers may use either plasma triglyceride values or an appropriate measure of body condition for assessing health in large sharks. PMID:27293643

  12. 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.

  13. 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

  14. 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.

  15. 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.

  16. 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.

  17. Boolean network model predicts knockout mutant phenotypes of fission yeast.

    Directory of Open Access Journals (Sweden)

    Maria I Davidich

    Full Text Available BOOLEAN NETWORKS (OR: networks of switches are extremely simple mathematical models of biochemical signaling networks. Under certain circumstances, Boolean networks, despite their simplicity, are capable of predicting dynamical activation patterns of gene regulatory networks in living cells. For example, the temporal sequence of cell cycle activation patterns in yeasts S. pombe and S. cerevisiae are faithfully reproduced by Boolean network models. An interesting question is whether this simple model class could also predict a more complex cellular phenomenology as, for example, the cell cycle dynamics under various knockout mutants instead of the wild type dynamics, only. Here we show that a Boolean network model for the cell cycle control network of yeast S. pombe correctly predicts viability of a large number of known mutants. So far this had been left to the more detailed differential equation models of the biochemical kinetics of the yeast cell cycle network and was commonly thought to be out of reach for models as simplistic as Boolean networks. The new results support our vision that Boolean networks may complement other mathematical models in systems biology to a larger extent than expected so far, and may fill a gap where simplicity of the model and a preference for an overall dynamical blueprint of cellular regulation, instead of biochemical details, are in the focus.

  18. Increased hepatotoxicity of acetaminophen in Hsp70i knockout mice

    International Nuclear Information System (INIS)

    The effect of the inducible forms of 70 kDa heat shock protein (Hsp70i) on acetaminophen (APAP) hepatotoxicity was assessed in an Hsp70i knockout mouse model. Absence of the Hsp70i protein in liver was verified by monitoring Hsp levels in knockout and control mice after heat stress (41.5 oC water bath immersion for 30 min). Hsp70i knockout mice were more susceptible to APAP-induced hepatotoxicity than controls, as indicated by elevated serum alanine aminotransferase activities 24 and 48 h after the APAP dose. Increased APAP hepatotoxicity in knockout mice was verified by morphological evaluation of liver sections. The difference in toxic response to APAP between knockout and control strain mice could not be attributed to differences in APAP bioactivation, assessed by measurement of CYP2E1 and glutathione S-transferase activities, hepatic nonprotein sulfhydryl content, or covalent binding of reactive APAP metabolites to proteins. Pretreatment with transient hyperthermia to produce a general upregulation of Hsps resulted in decreased APAP hepatotoxicity in both the knockout and control strains. Among thermally-pretreated mice, hepatotoxicity of APAP was greater in the knockouts compared with the control strain. These observations suggest that increased Hsp70i expression in response to APAP acts to limit the extent of tissue injury. Results further suggest that other factors related to heat stress can also contribute to protection against APAP toxicity

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

    International Nuclear Information System (INIS)

    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

  20. Seasonal Variation of Organogenetic Activity and Reserves Allocation in the Shoot Apex of Pinus pinaster Ait.

    OpenAIRE

    JORDY, MARIE‐NOËLLE

    2004-01-01

    • Background and aims To understand better the basic growth characteristics of pines and the fundamental properties of the shoot apical meristem (SAM), variations within the shoot apex of buds were studied.

  1. 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.

  2. 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.

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

    OpenAIRE

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

    2008-01-01

    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 ...

  4. Facile Preparation of a Platinum Silicide Nanoparticle-Modified Tip Apex for Scanning Kelvin Probe Microscopy

    OpenAIRE

    Lin, Chun-Ting; Chen, Yu-Wei; Su, James; Wu, Chien-Ting; Hsiao, Chien-Nan; Shiao, Ming-Hua; Chang, Mao-Nan

    2015-01-01

    In this study, we propose an ultra-facile approach to prepare a platinum silicide nanoparticle-modified tip apex (PSM tip) used for scanning Kelvin probe microscopy (SKPM). We combined a localized fluoride-assisted galvanic replacement reaction (LFAGRR) and atmospheric microwave annealing (AMA) to deposit a single platinum silicide nanoparticle with a diameter of 32 nm on the apex of a bare silicon tip of atomic force microscopy (AFM). The total process was completed in an ambient environment...

  5. 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.

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

    OpenAIRE

    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....

  7. 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...

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

    OpenAIRE

    Popp, C; Brunner, Dominik; DAMM Alexander; M. Van Roozendael; C. Fayt; B. Buchmann

    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 day in June 2010. N...

  9. 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

  10. 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.

  11. 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.

  12. Gomafu lncRNA knockout mice exhibit mild hyperactivity with enhanced responsiveness to the psychostimulant methamphetamine

    Science.gov (United States)

    Ip, Joanna Y.; Sone, Masamitsu; Nashiki, Chieko; Pan, Qun; Kitaichi, Kiyoyuki; Yanaka, Kaori; Abe, Takaya; Takao, Keizo; Miyakawa, Tsuyoshi; Blencowe, Benjamin J.; Nakagawa, Shinichi

    2016-01-01

    The long noncoding RNA Gomafu/MIAT/Rncr2 is thought to function in retinal cell specification, stem cell differentiation and the control of alternative splicing. To further investigate physiological functions of Gomafu, we created mouse knockout (KO) model that completely lacks the Gomafu gene. The KO mice did not exhibit any developmental deficits. However, behavioral tests revealed that the KO mice are hyperactive. This hyperactive behavior was enhanced when the KO mice were treated with the psychostimulant methamphetamine, which was associated with an increase in dopamine release in the nucleus accumbens. RNA sequencing analyses identified a small number of genes affected by the deficiency of Gomafu, a subset of which are known to have important neurobiological functions. These observations suggest that Gomafu modifies mouse behavior thorough a mild modulation of gene expression and/or alternative splicing of target genes. PMID:27251103

  13. 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

  14. 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.

  15. 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

  16. 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.

  17. Development-related PcG target in the apex 4 controls leaf margin architecture in Arabidopsis thaliana.

    Science.gov (United States)

    Engelhorn, Julia; Reimer, Julia J; Leuz, Iris; Göbel, Ulrike; Huettel, Bruno; Farrona, Sara; Turck, Franziska

    2012-07-01

    In a reverse genetics screen based on a group of genes enriched for development-related Polycomb group targets in the apex (DPAs), we isolated DPA4 as a novel regulator of leaf margin shape. T-DNA insertion lines in the DPA4 locus display enhanced leaf margin serrations and enlarged petals, whereas overexpression of DPA4 results in smooth margins. DPA4 encodes a putative RAV (Related to ABI3/VP1) transcriptional repressor and is expressed in the lateral organ boundary region and in the sinus of leaf serrations. DPA4 expression domains overlap with those of the known leaf shape regulator CUP-SHAPED COTYLEDON 2 (CUC2) and we provide evidence that DPA4 negatively regulates CUC2 expression independently of MIR164A, an established regulator of CUC2. Taken together, the data suggest DPA4 as a newly identified player in the signalling network that controls leaf serrations in Arabidopsis thaliana. PMID:22675210

  18. 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

  19. 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

  20. 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...

  1. 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

  2. P2X6 Knockout Mice Exhibit Normal Electrolyte Homeostasis

    Science.gov (United States)

    Viering, Daan H. H. M.; Bos, Caro; Bindels, René J. M.; Hoenderop, Joost G. J.

    2016-01-01

    ATP-mediated signaling is an important regulator of electrolyte transport in the kidney. The purinergic cation channel P2X6 has been previously localized to the distal convoluted tubule (DCT), a nephron segment important for Mg2+ and Na+ reabsorption, but its role in ion transport remains unknown. In this study, P2x6 knockout (P2x6-/-) mice were generated to investigate the role of P2X6 in renal electrolyte transport. The P2x6-/- animals displayed a normal phenotype and did not differ physiologically from wild type mice. Differences in serum concentration and 24-hrs urine excretion of Na+, K+, Mg2+ and Ca2+ were not detected between P2x6+/+, P2x6+/- and P2x6-/- mice. Quantitative PCR was applied to examine potential compensatory changes in renal expression levels of other P2x subunits and electrolyte transporters, including P2x1-5, P2x7, Trpm6, Ncc, Egf, Cldn16, Scnn1, Slc12a3, Slc41a1, Slc41a3, Cnnm2, Kcnj10 and Fxyd2. Additionally, protein levels of P2X2 and P2X4 were assessed in P2x6+/+ and P2x6-/- mouse kidneys. However, significant changes in expression were not detected. Furthermore, no compensatory changes in gene expression could be demonstrated in heart material isolated from P2x6-/- mice. Except for a significant (P<0.05) upregulation of P2x2 in the heart of P2x6-/- mice compared to the P2x6+/+ mice. Thus, our data suggests that purinergic signaling via P2X6 is not significantly involved in the regulation of renal electrolyte handling under normal physiological conditions. PMID:27254077

  3. 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

  4. 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

  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. Post-radiotherapy prostate biopsies reveal heightened apex positivity relative to other prostate regions sampled

    International Nuclear Information System (INIS)

    Background and purpose: Prostate biopsy positivity after radiotherapy (RT) is a significant determinant of eventual biochemical failure. We mapped pre- and post-treatment tumor locations to determine if residual disease is location-dependent. Materials and methods: There were 303 patients treated on a randomized hypofractionation trial. Of these, 125 underwent prostate biopsy 2-years post-RT. Biopsy cores were mapped to a sextant template, and 86 patients with both pre-/post-treatment systematic sextant biopsies were analyzed. Results: The pretreatment distribution of positive biopsy cores was not significantly related to prostate region (base, mid, apex; p = 0.723). Whereas all regions post-RT had reduced positive biopsies, the base was reduced to the greatest degree and the apex the least (p = 0.045). In 38 patients who had a positive post-treatment biopsy, there was change in the rate of apical positivity before and after treatment (76 vs. 71%; p = 0.774), while significant reductions were seen in the mid and base. Conclusion: In our experience, persistence of prostate tumor cells after RT increases going from the base to apex. MRI was used in planning and image guidance was performed daily during treatment, so geographic miss of the apex is unlikely. Nonetheless, the pattern observed suggests that attention to apex dosimetry is a priority

  8. 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.

  9. 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

  10. Rbpj conditional knockout reveals distinct functions of Notch4/Int3 in mammary gland development and tumorigenesis

    OpenAIRE

    Raafat, A.; Lawson, S; Bargo, S; Klauzinska, M; Strizzi, L; Goldhar, AS; Buono, K; Salomon, D.; Vonderhaar, BK; Callahan, R

    2008-01-01

    Transgenic mice expressing the Notch 4 intracellular domain (ICD) (Int3) in the mammary gland have two phenotypes: arrest of mammary alveolar/lobular development and mammary tumorigenesis. Notch4 signaling is mediated primarily through the interaction of Int3 with the transcription repressor/activator Rbpj. We have conditionally ablated the Rbpj gene in the mammary glands of mice expressing whey acidic protein (Wap)-Int3. Interestingly, Rbpj knockout mice (Wap-Cre+/Rbpj−/−/ Wap-Int3) have nor...

  11. 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...

  12. The effect of apex-finding errors on factor images obtained from factor analysis and oblique transformation

    International Nuclear Information System (INIS)

    The use of an oblique transformation to obtain physiological factors from a factor analysis of dynamic studies has recently been suggested. The technique involves using an apex-seeking routine to enable these factors and their contribution images to be formed. This paper investigates the effects of errors in apex-finding on these factor images. In particular it is shown that, apart from scaling, the factor image obtained from a given apex is independent of the position of that apex (and hence is independent of its own factor). (author)

  13. p21WAF1/Cip1/Sdi1 knockout mice respond to doxorubicin with reduced cardiotoxicity

    International Nuclear Information System (INIS)

    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 p21WAF1/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, IFNγ and TNFα 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: ► Doxorubicin induces p21 elevation in the myocardium. ► Doxorubicin causes dilated cardiomyopathy in wild type mice. ► p21 Knockout mice are resistant against doxorubicin induced cardiomyopathy. ► Lack of inflammatory response correlates with the resistance in p21 knockout mice.

  14. 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.

  15. Roles of hepatic glucokinase in intertissue metabolic communication: Examination of novel liver-specific glucokinase knockout mice.

    Science.gov (United States)

    Hayashi, Hirofumi; Sato, Yoshifumi; Li, Zhenghua; Yamamura, Ken-ichi; Yoshizawa, Tatsuya; Yamagata, Kazuya

    2015-05-01

    Glucokinase is expressed principally in pancreatic β-cells and hepatocytes, and catalyzes the phosphorylation of glucose to glucose-6-phosphate, a rate-limiting step of glycolysis. To better understand the roles of hepatic glucokinase, we generated Gck knockout mice by ablating liver-specific exon 1b. The knockout mice exhibited impaired glucose tolerance, decreased hepatic glycogen content, and reduced Pklr and Fas gene expression in the liver, indicating that hepatic glucokinase plays important roles in glucose metabolism. It has also been reported that hepatic glucokinase regulates the expression of thermogenesis-related genes in brown adipose tissue (BAT) and insulin secretion in response to glucose. However, the liver-specific Gck knockout mice displayed neither altered expression of thermogenesis-related genes in BAT nor impaired insulin secretion by β-cells under a normal chow diet. These results suggest that chronic suppression of hepatic glucokinase has a small influence on intertissue (liver-to-BAT as well as liver-to-β-cell) metabolic communication. PMID:25817793

  16. Knockout and transgenic mice of Trp53: what have we learned about p53 in breast cancer?

    International Nuclear Information System (INIS)

    The human p53 tumor suppressor gene TP53 is mutated at a high frequency in sporadic breast cancer, and Li-Fraumeni syndrome patients who carry germline mutations in one TP53 allele have a high incidence of breast cancer. In the 10 years since the first knockout of the mouse p53 tumor suppressor gene (designated Trp53) was published, much has been learned about the contribution of p53 to biology and tumor suppression in the breast through the use of p53 transgenic and knockout mice. The original mice deficient in p53 showed no mammary gland phenotype. However, studies using BALB/c-Trp53-deficient mice have demonstrated a delayed involution phenotype and a mammary tumor phenotype. Together with other studies of mutant p53 transgenes and p53 bitransgenics, a greater understanding has been gained of the role of p53 in involution, of the regulation of p53 activity by hormones, of the effect of mouse strain and modifier genes on tumor phenotype, and of the cooperation between p53 and other oncogenic pathways, chemical carcinogens and hormonal stimulation in mammary tumorigenesis. Both p53 transgenic and knockout mice are important in vivo tools for understanding breast cancer, and are yet to be exploited for developing therapeutic strategies in breast cancer

  17. 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.

  18. In-Flight Spectral Calibration of the APEX Imaging Spectrometer Using Fraunhofer Lines

    Science.gov (United States)

    Kuhlmann, Gerrit; Hueni, Andreas; Damm, Aalexander; Brunner, Dominik

    2015-06-01

    The Airborne Prism EXperiment (APEX) is an imaging spectrometer which allows to observe atmospheric trace gases such as nitrogen dioxide (NO2). Using a high resolution spectrum of solar Fraunhofer lines, APEX measurements collected during flight have been spectrally calibrated for centre wavelength positions (CW) and instrument slit function (ISF) and compared to the laboratory calibration. We find that CWs depend strongly on both across- and along-track position due to spectral smile and CWs dependency on ambient pressure. The width of the ISF is larger than estimated from the laboratory calibration but can be described by a linear scaling of the laboratory values. The ISF width depends on across but not on along-track direction. The results demonstrate the importance of characterizing and monitoring the instrument performance during flight and will be used to improve the Empa APEX NO2 retrieval algorithm.

  19. Rapid Development of Database Interfaces with Oracle APEX, used for the Controls Systems at CERN

    CERN Document Server

    Zaharieva, Z

    2009-01-01

    The need to rapidly prototype, build and deploy applications and to be able to react immediately to the changing user requirements is a challenge facing every enterprise. CERN – the largest particle research centre in the world – has tremendous data storage requirements, encompassing many different databases and has to quickly provide interfaces to visualize the data. This article will cover how Oracle APEX has been used to build several different database-centric interfaces related to the accelerator complex. Real-world applications will be discussed and it will be shown how Oracle APEX has met the preliminary requirements of the application developers and the user community at CERN. The article will address the question of when APEX could be a suitable choice of application development technology, and will share a developer's first-hand experience of both the good and bad points.

  20. 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.

  1. A new apex-ejecting perfused rat heart preparation: relation between coronary flow and loading conditions.

    Science.gov (United States)

    Wikman-Coffelt, J; Coffelt, R J; Rapcsak, M; Sievers, R; Rouleau, J L; Parmley, W W

    1983-12-01

    The isolated perfused rat heart is an important experimental preparation for both mechanical and biochemical studies. In order to define better the relationship between coronary flow and loading conditions, a new preparation was developed in which the left ventricle ejected through the apex, while the aortic perfusion pressure could be separately controlled at a higher level than the apex afterload. Results were compared with a standard aortic perfused and ejecting preparation. All analyses were made at low calcium concentration (1.6 mmol X litre-1) for reducing cardiac performance. Coronary flow was related to perfusion pressure in the aortic ejecting preparation when the aortic afterload chamber was between 6.0 and 9.3 kPa (45 and 70 mmHg). Coronary autoregulation was demonstrable in the apex ejecting preparation irrespective of the height of the apex afterload chamber and the aortic ejecting preparation when the aortic chamber was between 11.0 and 16.0 kPa (83 and 120 mmHg). Following the addition of 10(-6) mol X litre-1 adenosine, there was significant coronary vasodilatation, and flow became pressure dependent in all cases. In the apex-ejecting preparation, with a high aortic pressure, coronary flow remained at relatively fixed level, and increases in oxygen demand were met by increasing oxygen extraction. Thus, in this preparation oxygen extraction was directly related to workload. With abrupt increases in afterload, going from 6.0 to 9.3 kPa (45 to 70 mmHg) to a higher level, there was evidence of transient hypoxia with the aortic ejecting but not the apex ejecting preparation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:6661747

  2. 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

  3. 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

  4. 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...

  5. 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...

  6. Positional relationship between the maxillary sinus floor and the apex of the maxillary first molar using cone beam computed tomograph

    International Nuclear Information System (INIS)

    To assess the positional relationship between the maxillary sinus floor and the apex of the maxillary first molar using cone beam computed tomograph (CBCT). CBCTs from 127 subjects were analysed. A total of 134 maxillary first molars were classified according to their vertical and horizontal positional relationship to the maxillary sinus floor and measured according to the distance between the maxillary sinus floor and the maxillary first molar. Type III (The root projected laterally on the sinus cavity but its apex is outside the sinus boundaries) was dominated between 10 and 19 years and type I (The root apex was not in contact with the cortical borders of the sinus) was dominated (P<0.05) between 20 and 72 years on the vertical relationship between the maxillary sinus floor and the apex of the maxillary first molar. The maxillary sinus floor was located more at the apex (78.2%) than at the furcation (21.3%) for the palatal root. The distance from the root apex to the maxillary sinus floor confined to type I was increased according to the ages (P<0.05). Type M (The maxillary sinus floor was located between the buccal and the palatal root) was most common (72.4%) on the horizontal relationship between the maxillary sinus floor and the apex of the maxillary first molar. CBCT can provide highly qualified images for the maxillary sinus floor and the root apex of the maxillary first molar.

  7. Cardiomyocyte-specific conditional knockout of the histone chaperone HIRA in mice results in hypertrophy, sarcolemmal damage and focal replacement fibrosis

    Directory of Open Access Journals (Sweden)

    Nicolas Valenzuela

    2016-03-01

    Full Text Available 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.

  8. 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

  9. Knockout confirmation for Hurries: rapid genotype identification of Trypanosoma cruzi transfectants by polymerase chain reaction directly from liquid culture

    Directory of Open Access Journals (Sweden)

    Monica Visnieski Alcantara

    2014-07-01

    Full Text Available Gene knockout is a widely used approach to evaluate loss-of-function phenotypes and it can be facilitated by the incorporation of a DNA cassette having a drug-selectable marker. Confirmation of the correct knockout cassette insertion is an important step in gene removal validation and has generally been performed by polymerase chain reaction (PCR assays following a time-consuming DNA extraction step. Here, we show a rapid procedure for the identification of Trypanosoma cruzi transfectants by PCR directly from liquid culture - without prior DNA extraction. This simple approach enabled us to generate PCR amplifications from different cultures varying from 106-108 cells/mL. We also show that it is possible to combine different primer pairs in a multiplex detection reaction and even to achieve knockout confirmation with an extremely simple interpretation of a real-time PCR result. Using the “culture PCR” approach, we show for the first time that we can assess different DNA sequence combinations by PCR directly from liquid culture, saving time in several tasks for T. cruzi genotype interrogation.

  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. 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…

  12. 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...

  13. 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.

  14. 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.

  15. 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

  16. 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...

  17. APEX: A Computerized Simulation Game as the Basis for an Undergraduate Interdisciplinary Course.

    Science.gov (United States)

    Tannenbaum, Robert S.

    APEX is a computerized gaming simulation; it is also the name of an interdisciplinary course in environmental problems in urban areas introduced at the School of Health Science, Hunter College of the City University of New York. In the course, students assume the roles of decision makers in both the private and public sectors. They receive data…

  18. APEX (Air Pollution Exercise) Volume 9: Industrialist's Manual No. 5, Caesar's Rendering Plant.

    Science.gov (United States)

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

    The Industrialist's Manual No. 5, Caesar's Rendering Plant 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,…

  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. 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

  1. 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

  2. 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. PMID:25762293

  3. 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.

  4. 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

  5. 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.

  6. 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

  7. 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,...

  8. 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...

  9. 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.)

  10. 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...

  11. 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...

  12. 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...

  13. 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

  14. 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.

  15. 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.

  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. 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.

  18. Pregnane X receptor knockout mice display aging-dependent wearing of articular cartilage.

    Science.gov (United States)

    Azuma, Kotaro; Casey, Stephanie C; Urano, Tomohiko; Horie-Inoue, Kuniko; Ouchi, Yasuyoshi; Blumberg, Bruce; Inoue, Satoshi

    2015-01-01

    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. PMID:25749104

  19. Evaluation of Dentaport ZX and Raypex 6 electronic apex locators: An in vivo study

    OpenAIRE

    Moscoso, Saddy; Pineda, Kenneth; Basilio, Juan; Alvarado, Carlos; Roig Cayón, Miguel; Duran Sindreu, Fernando

    2013-01-01

    Med Oral Patol Oral Cir Bucal. 2014 Mar 1;19 (2):e202-5. Evaluation of Dentaport ZX and Raypex 6: An in vivo study e202 Journal section: Clinical and Experimental Dentistry Publication Types: Research Evaluation of Dentaport ZX and Raypex 6 electronic apex locators: An in vivo study Saddy Moscoso 1 , Kenneth Pineda 1 , Juan Basilio 1 , Carlos Alvarado 2 , Miguel Roig 2 , Fernando Duran- Sindreu 3 1 MD, Department of Restorative...

  20. Displacements of the organ of Corti by gel injections into the cochlear apex

    OpenAIRE

    Salt, Alec N.; Brown, Daniel J; Hartsock, Jared J.; Plontke, Stefan K

    2009-01-01

    In order to transduce sounds efficiently, the stereocilia of hair cells in the organ of Corti must be positioned optimally. Mechanical displacements, such as pressure differentials across the organ caused by endolymphatic hydrops, may impair sensitivity. Studying this phenomenon has been limited by the technical difficulty of inducing sustained displacements of stereocilia in vivo. We have found that small injections (0.5 to 2 μL) of Healon gel into the cochlear apex of guinea pigs produced s...

  1. 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.

  2. 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.

  3. Avulsion of permanent teeth with open apex: a systematic review of the literature

    OpenAIRE

    Felipe G. Belladonna; Ane Poly; João M. S. Teixeira; Viviane D. M. A. Nascimento; Sandra R. Fidel; Rivail A. S. Fidel

    2012-01-01

    Introduction: Considered the most serious of dental in Considered the most serious of dental injuries, uries,avulsion is known as the total displacement of tooth out of its socket. Treatment includes immediate replantation and its success is directly related to several factors. Objective: This paper aimed to review the literature in a systematic way on dental avulsion of in a systematic way on dental avulsion of permanent teeth with open apex, covering various topics such as: reason for av...

  4. Interdisciplinary Approach to a Tooth with Open Apex and Persistent Sinus

    OpenAIRE

    Das, Anoop N.; Krishnamohan Geetha; Ajay Varghese Kurian; Radhakrishnan Nair; Nandakumar, K

    2015-01-01

    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 ...

  5. The APEX\\/EPOS Quandary the Way Out via Low Energy Studies

    CERN Document Server

    Griffin, J J

    1998-01-01

    A scorecard summary of the various data of the ``Sharp Lepton Problem'' is presented. The present situation, in which APEX reports ``...no evidence for sharp pairs...'' even as their data exhibits a sharp pair excess near 800 keV, is discussed. Two kinds of low energy experiments utilizing non-heavy ion processes are suggested as means to break the impasse arising from the ambiguity of the present heavy ion data.

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

    Science.gov (United States)

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

    2012-09-01

    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 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.

  7. 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.

  8. 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

  9. 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...

  10. 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.

  11. 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

  12. 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.

  13. 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 ...

  14. 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...

  15. 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...

  16. Treatment with anti-interferon-gamma monoclonal antibodies modifies experimental autoimmune encephalomyelitis in interferon-gamma receptor knockout mice

    DEFF Research Database (Denmark)

    Espejo, C; Penkowa, M; Sáez-Torres, I;

    2001-01-01

    The role of interferon-gamma (IFN-gamma) in the pathogenesis of multiple sclerosis and experimental autoimmune encephalomyelitis (EAE) is still controversial. We have studied the function of IFN-gamma and its receptor in the EAE model using two different IFN-gamma receptor knockout (IFN-gamma R......(-/-)) mouse types: C57Bl/6x129Sv, with a disruption of the IFN-gamma receptor cytoplasmic domain, and 129Sv, homozygous for a disrupted IFN-gamma receptor gene. Mice were immunized with peptide 40-55 from rat myelin oligodendrocyte glycoprotein. A subgroup of mice was treated with anti-IFN-gamma monoclonal...

  17. 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...

  18. 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.

  19. Hippocampal dysfunction in the Euchromatin histone methyltransferase 1 heterozygous knockout mouse model for Kleefstra syndrome.

    Science.gov (United States)

    Balemans, Monique C M; Kasri, Nael Nadif; Kopanitsa, Maksym V; Afinowi, Nurudeen O; Ramakers, Ger; Peters, Theo A; Beynon, Andy J; Janssen, Sanne M; van Summeren, Rik C J; Eeftens, Jorine M; Eikelenboom, Nathalie; Benevento, Marco; Tachibana, Makoto; Shinkai, Yoichi; Kleefstra, Tjitske; van Bokhoven, Hans; Van der Zee, Catharina E E M

    2013-03-01

    Euchromatin histone methyltransferase 1 (EHMT1) is a highly conserved protein that catalyzes mono- and dimethylation of histone H3 lysine 9, thereby epigenetically regulating transcription. Kleefstra syndrome (KS), is caused by haploinsufficiency of the EHMT1 gene, and is an example of an emerging group of intellectual disability (ID) disorders caused by genes encoding epigenetic regulators of neuronal gene activity. Little is known about the mechanisms underlying this disorder, prompting us to study the Euchromatin histone methyltransferase 1 heterozygous knockout (Ehmt1(+/-)) mice as a model for KS. In agreement with the cognitive disturbances observed in patients with KS, we detected deficits in fear extinction learning and both novel and spatial object recognition in Ehmt1(+/-) mice. These learning and memory deficits were associated with a significant reduction in dendritic arborization and the number of mature spines in hippocampal CA1 pyramidal neurons of Ehmt1(+/-) mice. In-depth analysis of the electrophysiological properties of CA3-CA1 synapses revealed no differences in basal synaptic transmission or theta-burst induced long-term potentiation (LTP). However, paired-pulse facilitation (PPF) was significantly increased in Ehmt1(+/-) neurons, pointing to a potential deficiency in presynaptic neurotransmitter release. Accordingly, a reduction in the frequency of miniature excitatory post-synaptic currents (mEPSCs) was observed in Ehmt1(+/-) neurons. These data demonstrate that Ehmt1 haploinsufficiency in mice leads to learning deficits and synaptic dysfunction, providing a possible mechanism for the ID phenotype in patients with KS. PMID:23175442

  20. 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

  1. 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

  2. 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

  3. 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

  4. 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

  5. Characteristics of Skeletal Muscle Fibers of SOD1 Knockout Mice

    OpenAIRE

    Hiroshi Nagahisa; Kazuma Okabe; Yoshihito Iuchi; Junichi Fujii; Hirofumi Miyata

    2016-01-01

    Cu/Zn superoxide dismutase (SOD1) knockout (KO) mice are known as an aging model in some aspects, but the damage and regeneration process of each fiber type have not been sufficiently studied. In this study, we investigated the damage and satellite cell state of the gastrocnemius muscle in SOD1 KO mice (6 months old) using immunohistochemical staining and real-time RT-PCR. The proportion of central nuclei-containing Type IIx/b fibers in the deep and superficial portions of the gastrocnemius m...

  6. Electron-induced neutron knockout from 4He

    International Nuclear Information System (INIS)

    The differential cross section for electron-induced neutron knockout in the reaction 4He(e,e'n)3He has been measured for the first time with a statistical accuracy of 11%. The experiment was performed in quasielastic kinematics at a momentum transfer of 300 MeV/c and in the missing-momentum range of 25-70 MeV/c. The comparison of the data with theoretical calculations shows an impressive increase of the cross section resulting from final state interaction effects. Specifically, the p-n charge-exchange process dominates the cross section in this kinematical regime

  7. 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.

  8. Thyrotoropin receptor knockout changes monoaminergic neuronal system and produces methylphenidate-sensitive emotional and cognitive dysfunction.

    Science.gov (United States)

    Mouri, Akihiro; Hoshino, Yuta; Narusawa, Shiho; Ikegami, Keisuke; Mizoguchi, Hiroyuki; Murata, Yoshiharu; Yoshimura, Takashi; Nabeshima, Toshitaka

    2014-10-01

    Attention deficit/hyperactivity disorder (ADHD) has been reported in association with resistance to thyroid hormone, a disease caused by a mutation in the thyroid hormone receptor β (TRβ) gene. TRβ is a key protein mediating down-regulation of thyrotropin (TSH) expression by 3,3',5-tri-iodothyronine (T3), an active form of thyroid hormone. Dysregulation of TSH and its receptor (TSHR) is implicated in the pathophysiology of ADHD but the role of TSHR remains elusive. Here, we clarified a novel role for TSHR in emotional and cognitive functions related to monoaminergic nervous systems. TSHR knockout mice showed phenotypes of ADHD such as hyperactivity, impulsiveness, a decrease in sociality and increase in aggression, and an impairment of short-term memory and object recognition memory. Administration of methylphenidate (1, 5 and 10mg/kg) reversed impulsiveness, aggression and object recognition memory impairment. In the knockout mice, monoaminergic changes including decrease in the ratio of 3-methoxy-4-hydroxyphenylglycol/noradrenaline and increase in the ratio of homovanillic acid/dopamine were observed in some brain regions, accompanied by increase in the expression of noradrenaline transporter in the frontal cortex. When TSH was completely suppressed by the supraphysiological administration of T3 to the adult mice, some behavioral and neurological changes in TSHR KO mice were also observed, suggesting that these changes were not due to developmental hypothyroidism induced by the inactivation of TSHR but to the loss of the TSH-TSHR pathway itself. Taken together, the present findings suggest a novel role for TSHR in behavioral and neurological phenotypes of ADHD. PMID:25016105

  9. 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.

  10. 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

  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. Post-operative high dose rate vaginal apex brachytherapy in stage I endometrial adenocarcinoma

    International Nuclear Information System (INIS)

    Purpose/Objective: Patients with Stage I endometrial adenocarcinoma have traditionally been treated with total abdominal hysterectomy/bilateral salpingo-oophorectomy and radiation. The reported incidence of local recurrence in surgically treated patients with FIGO Stage IA or IB endometrial adenocarcinoma is 4-10% at 2 years. Combined surgery and radiation has resulted in a reduction of recurrence to 2-6%. We report the presentation, actuarial survival, actuarial rate of local failure, salvage rate, and complications for patients undergoing high dose rate (HDR) vaginal apex brachytherapy following surgery. Materials and Methods: Between 1985 and 1994 a total of 286 patients with FIGO Stage I endometrioid uterine adenocarcinoma were treated with HDR Ir-192 vaginal apex brachytherapy alone to a total dose of 21 Gy in 3 fractions at 0.5 cm from the vaginal mucosa. The pathologic stage by treatment group was IA: 31%, IB: 68%, and IC: 1%. The histologic grade of the patient's tumors was grade 1: 69%, 2: 29%, and 3: 2% of patients. The median time from surgery to radiation was 34 days (range 14-66 days). The median follow-up for 286 patients with Stage IA (92 patients), IB (190 patients), and IC (4 patients) was respectively, 37, 35 and 40 months (2 patients lost to follow-up prior to 6 months). Results: Patients presented with vaginal bleeding (94%) or abnormal pap smear (6%) at a median age for Stage IA and IB, of 54 and 63 years, respectively (range 32-88). The 5-year overall actuarial survival rate was 94.5%. The 5-year actuarial survival rate by histologic grade was 97.5% and 91.5% for FIGO grade 1 and 2, respectively (p=.011). The 5-year actuarial survival rate by depth of myometrial invasion was 99.0% and 92.5% for Stage IA and IB, respectively (p=.029). Median overall time to failure is 19.5 months (range 10-36 months). The 5-year actuarial rate of local failure was 4.5%. The overall failure rate in our study group was 2.8% (8 patients), local failure only 1

  13. 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.

  14. 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...

  15. 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.

  16. USM Apex University Status: Transforming Higher Education For A Sustainable Tomorrow

    OpenAIRE

    Abd Razak, Dzulkifli

    2009-01-01

    This special editorial for the month of January 2009 is “THE” interview with Professor Tan Sri Dato’ Dzulkifli Abdul Razak, Vice Chancellor of the Universiti Sains Malaysia (USM). He talked to us about the Apex University status that was conferred on 3rd September 2008. While most newspapers and the rest of the media were in frenzy interviewing him that month, MJMS decided to catch up with him on Monday 15th December 2008 just before the celebration at the Dewan Utama, Universiti Sains Malays...

  17. Cosmological studies of the Sunyaev-Zel'dovich Effect with the APEX-SZ instrument

    Science.gov (United States)

    Mehl, Jared Joseph

    We have built the APEX-SZ instrument to observe galaxy clusters with the Sunyaev-Zel'dovich Effect (SZE), in which Cosmic Microwave Background (CMB) photons are scattered off of the hot electron gas within a galaxy cluster. Galaxy clusters are the largest collapsed objects in the universe and make excellent tracers of the distribution and evolution of matter. Galaxy cluster surveys can be used to measure the matter density of the universe O m , the equation of state of dark energy w, the amplitude of matter fluctuations at 8 h -1 Mpc s 8 , and the Hubble constant H 0 . Galaxy cluster observations with the SZE also provide a unique probe of galaxy cluster physics. I will describe the design and fabrication of the 330-element Transition-Edge Sensor (TES) bolometer array used in APEX-SZ. CMB power is measured with an aluminum-titanium bilayer TES centered on a silicon-nitride spiderweb covered with a thin-film gold absorber. Frequency-domain multiplexing is used to read out the TESs. An integration cavity is formed behind a smooth-walled conical horn by the silicon wafer in direct contact with a metal backshort. This cavity couples the gold absorber to incoming millimeter-wave radiation. I will also describe finite element analysis electromagnetic simulations of the integration cavity intended to optimize the optical efficiency of the bolometer array. The full configuration APEX-SZ instrument was deployed to the 12-meter diameter APEX telescope sited in Chile in March 2007. Observations of the Bullet cluster, a massive galaxy cluster merger perpendicular to the line-of-sight, were made in August 2007. The Bullet cluster was detected in the SZE with a23s significance within the central 1' radius of the source position. We measure a temperature decrement of --880 ± 80mK CMB using an isothermal elliptical b- model fit to the temperature map. We measure a cluster gas mass fraction fg = 0.121±0.038 within a radius 1.42 Mpc, consistent with X-ray and weak lensing results

  18. 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

  19. Analysis of Base-Apex Lead Electrocardiogram in Clinically Healthy Kermani Sheep

    OpenAIRE

    Tajik, Javad; Amir Saeed SAMIMI; Shojaeepour, Saeedeh; JARAKANI, Sajad

    2016-01-01

    The normal electrocardiographic (ECG) parameters in the base-apex lead were evaluated in 40 clinically healthy Kermani sheep, and compared between sexes and three age groups. The heart rate varied from 83-192 beats/min with a mean of 128.9±4.7 beats/min (Mean±SEM). Sinus arrhythmia was the only observed cardiac dysrhythmia on the ECG traces diagnosed in 45% of animals. No significant difference was found in heart rate, amplitude, and duration of ECG waves and intervals between two sexes. Neve...

  20. A Survey of Jet Aircraft PM by TEM in APEX III

    Science.gov (United States)

    VanderWal, Randy L.; Bryg, Victoria M.

    2014-01-01

    Based upon field testing during the NASA led APEX III campaign conducted in November 2005 at the NASA Glenn Research Center in coordination with Continental Airlines and Cleveland Hopkins International Airport. This paper reports observations of particulate emissions collected from a suite of jet engine aircraft to assess differences and similarities in soot macro- micro- and nanostructure using transmission electron microscopy (TEM). Aggregates are compact, primary particle sizes varied and nanostructure mixed. Comparisons are made to more familiar laboratory flame-generated soot as a well-studied point of reference. Results are interpreted in terms of turbulence interacting with the different stages of particle formation and growth.

  1. 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.

  2. Controlling the apex angle of carbon cone induced by 1.2 keV argon ions at room temperature

    International Nuclear Information System (INIS)

    Carbon nanofiber-tipped-cones with controllable apex angle were fabricated by sputtering graphite with 1.2keV Ar+ ions in different incident angles at room temperature. The density of carbon cones was estimated at 1 x 109- 1 x 1010/cm2. The cones with carbon nanofiber were oriented to the ion-beam direction. By increasing the incidence from 30 degree to 60 degree, the apex angle of cones decreased from 33 degree to 20 degree, and the aspect ratio increased from 250nm/150nm to 1200nm/400nm. The decreased apex angle, and the increased aspect ratio and density of the cones, were attributed to decreasing effective diffuse coefficient induced by the ion beam and increasing sputtering yield. By increasing the current density from 200 μA/cm2 to 800 μA/cm2, the apex angle of cones decreased from 90 degree to 20 degree and the height of cones increased from 100nm to 1200nm. The increased dose rate caused by larger current densities of the ion beam should result in the different number of the sputtered atom or cluster, which was considered as reason of the decrement of the apex angle and the increment of the height of cones. (authors)

  3. Ethanol self-administration in serotonin transporter knockout mice: unconstrained demand and elasticity.

    Science.gov (United States)

    Lamb, R J; Daws, L C

    2013-10-01

    Low serotonin function is associated with alcoholism, leading to speculation that increasing serotonin function could decrease ethanol consumption. Mice with one or two deletions of the serotonin transporter (SERT) gene have increased extracellular serotonin. To examine the relationship between SERT genotype and motivation for alcohol, we compared ethanol self-administration in mice with zero (knockout, KO), one (HET) or two copies (WT) of the SERT gene. All three genotypes learned to self-administer ethanol. The SSRI, fluvoxamine, decreased responding for ethanol in the HET and WT, but not the KO mice. When tested under a progressive ratio schedule, KO mice had lower breakpoints than HET or WT. As work requirements were increased across sessions, behavioral economic analysis of ethanol self-administration indicated that the decreased breakpoint in KO as compared to HET or WT mice was a result of lower levels of unconstrained demand, rather than differences in elasticity, i.e. the proportional decreases in ethanol earned with increasing work requirements were similar across genotypes. The difference in unconstrained demand was unlikely to result from motor or general motivational factors, as both WT and KO mice responded at high levels for a 50% condensed milk solution. As elasticity is hypothesized to measure essential value, these results indicate that KO value ethanol similarly to WT or HET mice despite having lower break points for ethanol. PMID:23927813

  4. 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...

  5. 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

  6. Facile Preparation of a Platinum Silicide Nanoparticle-Modified Tip Apex for Scanning Kelvin Probe Microscopy.

    Science.gov (United States)

    Lin, Chun-Ting; Chen, Yu-Wei; Su, James; Wu, Chien-Ting; Hsiao, Chien-Nan; Shiao, Ming-Hua; Chang, Mao-Nan

    2015-12-01

    In this study, we propose an ultra-facile approach to prepare a platinum silicide nanoparticle-modified tip apex (PSM tip) used for scanning Kelvin probe microscopy (SKPM). We combined a localized fluoride-assisted galvanic replacement reaction (LFAGRR) and atmospheric microwave annealing (AMA) to deposit a single platinum silicide nanoparticle with a diameter of 32 nm on the apex of a bare silicon tip of atomic force microscopy (AFM). The total process was completed in an ambient environment in less than 3 min. The improved potential resolution in the SKPM measurement was verified. Moreover, the resolution of the topography is comparable to that of a bare silicon tip. In addition, the negative charges found on the PSM tips suggest the possibility of exploring the use of current PSM tips to sense electric fields more precisely. The ultra-fast and cost-effective preparation of the PSM tips provides a new direction for the preparation of functional tips for scanning probe microscopy. PMID:26471480

  7. 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...

  8. 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.

  9. 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.

  10. 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.

  11. 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.

  12. 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.

  13. Oyster polynuclear aromatic hydrocarbon fingerprinting applied to the Apex barge oil spill

    International Nuclear Information System (INIS)

    An estimated 692,000 gallons of catalytic feed stock oil was spilled into Galveston Bay on July 28, 1990, when a tanker collided with three Apex barges in the Houston Ship Channel. Oysters were collected and analyzed from Galveston Bay Todd's Dump (GBRD) before the spill and after the spill. Oysters were also collected from Galveston Bay Redfish Island (GBRI), a site known to be impacted by the spill, 37 and 110 days after the spill. The spilled oil was also analyzed. The concentration of 18 polynuclear aromatic hydrocarbons (PAH) measured as part of the National Oceanic and Atmospheric Administration's National Status ampersand Trends (NS ampersand T) showed a sharp increase from 100 ng/g to over 600 ng/g. Concentrations of these 18 PAHs were also found at GBRI. Fingerprinting techniques applied to data from oyster analyses demonstrated the presence of bioavailable Apex Barge oil 37, 110, and 132 days after the spill at GHTD and GBRI. Fingerprinting becomes less diagnostic with time due to possible environmental weathering of the oil. The fingerprint from GBTD 495 and 851 days after the spill will be presented and discussed

  14. Postoperative high dose rate vaginal apex brachytherapy in stage I endometrial adenocarcinoma

    International Nuclear Information System (INIS)

    Introduction: Patients with stage I endometrial adenocarcinoma have traditionally been treated with TAH/BSO and radiation. The reported incidence of local recurrence in surgically treated patients with Figo stage IA or IB endometrial adenocarcinoma is 4-10% at 2 years. Combined surgery and radiation has resulted in a reduction of recurrence to 2-6%. We report the presentation, local and distant control, salvage rate, survival and complications for patients undergoing high dose rate (HDR) vaginal apex brachytherapy following surgery. Methods: Between 1985 and 1994 a total of 302 patients with Figo stage I endometrial carcinoma were treated with HDR Ir-192 vaginal apex brachytherapy to a total dose of 21 Gy in 3 fractions at 0.5 cm from the vaginal mucosa. The pathologic stage by treatment group was IA: 31%, IB: 68%, and IC: 1%. The histologic grade of the patient's tumors was grade 1: 69%, 2: 27%, and 3: 4%. The median time from surgery to radiation was 33 days (range 14-66 days). The median follow-up for 300 patients with stage IA (91 patients), IB (205 patients), and IC (4 patients) was respectively 36, 34 and 40 months (2 patients lost to follow-up prior to 6 months). Results: Patients presented with vaginal bleeding (94%) or abnormal pap smear (6%) at a median age for stage IA and IB, of 55 and 64 years, respectively. The crude overall survival of the patient population at 2 years is 95%. Median overall time to failure is 19.5 months (range 10-36 months). The overall failure rate was 2.7% (8 patients), local failure only 1.0% (38% of failed group), distant failure only 0.3% (12% of failed group) and combined local/distant failure 1.3% (50% of failed group). The local failure rate for pathologic stage IA patients was 1.0% and no distant disease was observed. The local failure rate for pathologic stage IB patients was 3.4% 7/205 and distant failure was 2.4% 5/205. The majority of patients with recurrence had grade 2 histologic changes 5/8. The overall salvage rate

  15. Healing of apical rarefaction of three nonvital open apex anterior teeth using a white portland cement apical plug

    Directory of Open Access Journals (Sweden)

    Amitabha Chakraborty

    2012-01-01

    Full Text Available The major challenge of performing root canal treatment in an open apex pulp-less tooth is to obtain a good apical seal. MTA has been successfully used to achieve a good apical seal, wherein the root canal obturation can be done immediately. MTA and White Portland Cement has been shown similarity in their physical, chemical and biological properties and has also shown similar outcome when used in animal studies and human trials. In our study, open apex of three non vital upper central incisors has been plugged using modified white Portland cement. 3 to 6 months follow up revealed absence of clinical symptoms and disappearance of peri-apical rarefactions. The positive clinical outcome may encourage the future use of white Portland cement as an apical plug material in case of non vital open apex tooth as much cheaper substitute of MTA.

  16. Preparation of Knockout Extract by Immunoaffinity Column and Its Application

    Directory of Open Access Journals (Sweden)

    Osamu Morinaga

    2012-12-01

    Full Text Available Importance of herbal medicines have recently increased owing to rising interest in their health benefits. However, medicinal plant extracts are complex mixtures of phytochemicals that act synergistically or additively on specific and/or multiple molecular and cellular targets. Thus, it is difficult to examine the actual pharmacological roles of active compounds in plant extracts. This review describes a new strategy for isolating target compounds from plant extracts using immunoaffinity columns coupled with monoclonal antibodies (mAbs against natural compounds. Through one-step purification using mAb-coupled immunoaffinity columns, we succeeded in preparing a knockout (KO extract, which contains all components except the target compound. Furthermore, we investigated the pharmacological effects of the KO extract to reveal the actual effects of a bioactive compound in the crude extract. This approach may help determine the potential function of target compounds in herbal medicines.

  17. 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 not...

  18. 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 < 0.001). The trabecular bone structure...

  19. 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.

  20. 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

  1. Medium effects on spin observables of proton knockout reactions

    International Nuclear Information System (INIS)

    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

  2. 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.

  3. 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…

  4. 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.

  5. Atm heterozygous deficiency enhances development of mammary carcinomas in p53 heterozygous knockout mice

    International Nuclear Information System (INIS)

    Ataxia-telangiectasia is an autosomal-recessive disease that affects neuro-immunological functions, associated with increased susceptibility to malignancy, chromosomal instability and hypersensitivity to ionizing radiation. Although ataxia-telangiectasia mutated (ATM) heterozygous deficiency has been proposed to increase susceptibility to breast cancer, some studies have not found excess risk. In experimental animals, increased susceptibility to breast cancer is not observed in the Atm heterozygous deficient mice (Atm+/-) carrying a knockout null allele. In order to determine the effect of Atm heterozygous deficiency on mammary tumourigenesis, we generated a series of Atm+/- mice on the p53+/- background with a certain predisposition to spontaneous development of mammary carcinomas, and we examined the development of the tumours after X-irradiation. BALB/cHeA-p53+/- mice were crossed with MSM/Ms-Atm+/- mice, and females of the F1 progeny ([BALB/cHeA × MSM/Ms]F1) with four genotypes were used in the experiments. The mice were exposed to X-rays (5 Gy; 0.5 Gy/min) at age 5 weeks. We tested the effect of haploinsufficiency of the Atm gene on mammary tumourigenesis after X-irradiation in the p53+/- mice of the BALB/cHeA × MSM/Ms background. The singly heterozygous p53+/- mice subjected to X-irradiation developed mammary carcinomas at around 25 weeks of age, and the final incidence of mammary carcinomas at 39 weeks was 31% (19 out of 61). The introduction of the heterozygous Atm knockout alleles into the background of the p53+/- genotype significantly increased the incidence of mammary carcinoma to 58% (32 out of 55) and increased the average number of mammary carcinomas per mouse. However, introduction of Atm alleles did not change the latency of development of mammary carcinoma. Our results indicate a strong enhancement in mammary carcinogenesis by Atm heterozygous deficiency in p53+/- mice. Thus, doubly heterozygous mice represent a useful model system with which to

  6. 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.

  7. Healing of apical rarefaction of three nonvital open apex anterior teeth using a white portland cement apical plug

    OpenAIRE

    Amitabha Chakraborty; Bibhas Dey; Reema Dhar; Prabir Sardar

    2012-01-01

    The major challenge of performing root canal treatment in an open apex pulp-less tooth is to obtain a good apical seal. MTA has been successfully used to achieve a good apical seal, wherein the root canal obturation can be done immediately. MTA and White Portland Cement has been shown similarity in their physical, chemical and biological properties and has also shown similar outcome when used in animal studies and human trials. In our study, open apex of three non vital upper central incisors...

  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. Altered lipid metabolism in Hfe-knockout mice promotes severe NAFLD and early fibrosis.

    Science.gov (United States)

    Tan, Terrence C H; Crawford, Darrell H G; Jaskowski, Lesley A; Murphy, Therese M; Heritage, Mandy L; Subramaniam, V Nathan; Clouston, Andrew D; Anderson, Gregory J; Fletcher, Linda M

    2011-11-01

    The HFE protein plays a crucial role in the control of cellular iron homeostasis. Steatosis is commonly observed in HFE-related iron-overload disorders, and current evidence suggests a causal link between iron and steatosis. Here, we investigated the potential contribution of HFE mutations to hepatic lipid metabolism and its role in the pathogenesis of nonalcoholic fatty liver disease. Wild-type (WT) and Hfe knockout mice (Hfe(-/-)) were fed either standard chow, a monounsaturated low fat, or a high-fat, high-carbohydrate diet (HFD) and assessed for liver injury, body iron status, and markers of lipid metabolism. Despite hepatic iron concentrations and body weights similar to WT controls, Hfe(-/-) mice fed the HFD developed severe hypoxia-related steatohepatitis, Tnf-α activation, and mitochondrial respiratory complex and antioxidant dysfunction with early fibrogenesis. These features were associated with an upregulation in the expression of genes involved in intracellular lipid synthesis and trafficking, while transcripts for mitochondrial fatty acid β-oxidation and adiponectin signaling-related genes were significantly attenuated. In contrast, HFD-fed WT mice developed bland steatosis only, with no inflammation or fibrosis and no upregulation of lipogenesis-related genes. A HFD led to reduced hepatic iron in Hfe(-/-) mice compared with chow-fed mice, despite higher serum iron, decreased hepcidin expression, and increased duodenal ferroportin mRNA. In conclusion, our results demonstrate that Hfe(-/-) mice show defective hepatic-intestinal iron and lipid signaling, which predispose them toward diet-induced hepatic lipotoxicity, accompanied by an accelerated progression of injury to fibrosis. PMID:21817060

  10. 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.

  11. 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.

  12. Spontaneous fusion across the apex of severe thoracolumbar Scheuermann′s kyphosis: A surgical consideration

    Directory of Open Access Journals (Sweden)

    Tsirikos Athanasios

    2010-01-01

    Full Text Available Considerable debate exists regarding the pathogenesis, natural history and treatment of Scheuermann′s kyphosis. Surgical correction is indicated in the presence of severe kyphosis which carries the risk of neurological complications, persistent back pain and significant cosmetic deformity. This can be achieved through a posterior-only or an anteroposterior approach. Spontaneous fusion in association with Scheuermann′s kyphosis has not been previously described. This is an important consideration if surgical correction of the kyphosis is planned. Two patients with severe thoracolumbar Scheuermann′s kyphosis who developed spontaneous posterior and anteroposterior fusion across the apex of the deformity are presented. The surgical treatment and final outcome is discussed.

  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. 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; Hvidsten, Svend; Zebrowski, David C; Engel, Felix Benedikt; Sheikh, Søren Paludan

    2016-01-01

    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 be an...... intermediate response before the onset of complete heart regeneration. Myocardial damage was performed by AR in postnatal day 1 C57BL/6 mice, and myocardial function and scarring assessed at day 180 using F-18-fluorodeoxyglucose positron emission tomography (FDG-PET) and histology, respectively. AR mice...

  15. Disease-induced decline of an apex predator drives invasive dominated states and threatens biodiversity.

    Science.gov (United States)

    Hollings, Tracey; Jones, Menna; Mooney, Nick; McCallum, Hamish

    2016-02-01

    Apex predators are important in protecting biodiversity through top-down influence on food webs. Their loss is linked with competitive release of invasive mesopredators and species extinctions. The Tasmanian devil (Sarcophilus harrisii) has experienced severe declines over a 15-yr period as a novel transmissible cancer has spread across its current geographic range. We surveyed the mammalian community, using hair traps, across the spatial extent of the devil's progressive population decline. We found increased activity of alien invasive species (feral cats, black rats), and reduced small and medium-sized native prey species in response to the timing of the decline. In areas of long-term devil decline, invasive species comprised a significantly larger proportion of the community. The results provide evidence that the devil plays a keystone role in Tasmania's ecosystem with their decline linked to a shift toward an invasive state and biodiversity loss in one of Australia's most intact faunal communities. PMID:27145614

  16. 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.

  17. Balanced Waveguide HEB Mixer for APEX 1.3 THz receiver

    Science.gov (United States)

    Meledin, Denis; Pantaleev, Miroslav; Pavolotsky, Alexey; Risacher, Christophe; Perez Robles, Victor A.; Belitsky, Victor; Drakinskiy, Vladimir; Cherednichenko, Sergey

    2005-05-01

    We present results on the design and construction of a waveguide balanced Hot Electron Bolometer (HEB) Terahertz mixer for Atacama Pathfinder EXperiment (APEX), band T2 covering 1250-1390 GHz frequency range. In the proposed design, a waveguide balanced mixer is realized using a quadrature scheme. The two identical HEB elements are integrated with RF choke filters, DC-bias, IF circuitry, and fabricated from 4 nm thick NbN film deposited on a crystalline quartz substrate with dimensions of 1100 μm×70 μm × 17 μm. We have designed and fabricated an input 3 dB quadrature waveguide hybrid. For its fabrication, we use micromachining approach to achieve low insertion loss and symmetrical division of the RF and local oscillator (LO) power within the band of interest. We plan to use two HEB mixer configurations with different probe impedance values of 55, 70 Ohm within 1250-1390 GHz frequency range.

  18. Rovibrational effects on nuclear shielding of apex nuclei in bent molecules

    International Nuclear Information System (INIS)

    The isotope shifts of non-end nuclei with lone pairs are generally larger than those of nuclei without lone pairs. Using the bent triatomic molecule as a prototype, we examine the mass and temperature dependence of the mean bond angle deformation and the mean displacement along a bond due to centrifugal stretching and anharmonic vibration. We find that the temperature dependence of determines whether the temperature dependence of the shielding of the apex nucleus will be normal [(dsigma0/dT) in the hydrides can lead to opposing temperature effects on shielding while the mass effects lead to normal isotope shifts. We performed similar calculations for the trigonal pyramidal molecules in order to explain the observed abnormal temperature dependence of the 15N and 31P nuclear shielding in NH3 and PH3

  19. 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

  20. 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.

  1. 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.

  2. 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.

  3. Biotrophic Development of Sporisorium reilianum f. sp. zeae in Vegetative Shoot Apex of Maize.

    Science.gov (United States)

    Martinez, C; Roux, C; Dargent, R

    1999-03-01

    ABSTRACT Sporisorium reilianum f. sp. zeae is the causal agent of maize head smut, a disease present in several regions of France. A cytological study was carried out to describe a key step of the fungal etiology, in which the mycelium invades the vegetative shoot apex. Light and transmission electron microscopy observations show that the fungus is mostly intracellular and suggest that it passes through the host cell wall by lysis and mechanical pressure. The hyphae are surrounded by an amorphous vesicle-rich layer limited by a membrane related to the host plasmalemma. The encasement can be considered as an exchange zone between the plant and the fungus. The infected host cells appear normal; therefore, the fungus seems to act like a biotrophic endophyte. PMID:18944766

  4. 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

  5. Analysis of knockout mice suggests a role for VGF in the control of fat storage and energy expenditure

    Directory of Open Access Journals (Sweden)

    Chakraborty Tandra

    2009-10-01

    Full Text Available Abstract Background Previous studies of mixed background mice have demonstrated that targeted deletion of Vgf produces a lean, hypermetabolic mouse that is resistant to diet-, lesion-, and genetically-induced obesity. To investigate potential mechanism(s and site(s of action of VGF, a neuronal and endocrine secreted protein and neuropeptide precursor, we further analyzed the metabolic phenotypes of two independent VGF knockout lines on C57Bl6 backgrounds. Results Unlike hyperactive VGF knockout mice on a mixed C57Bl6-129/SvJ background, homozygous mutant mice on a C57Bl6 background were hypermetabolic with similar locomotor activity levels to Vgf+/Vgf+ mice, during day and night cycles, indicating that mechanism(s other than hyperactivity were responsible for their increased energy expenditure. In Vgf-/Vgf- knockout mice, morphological analysis of brown and white adipose tissues (BAT and WAT indicated decreased fat storage in both tissues, and decreased adipocyte perimeter and area in WAT. Changes in gene expression measured by real-time RT-PCR were consistent with increased fatty acid oxidation and uptake in BAT, and increased lipolysis, decreased lipogenesis, and brown adipocyte differentiation in WAT, suggesting that increased sympathetic nervous system activity in Vgf-/Vgf- mice may be associated with or responsible for alterations in energy expenditure and fat storage. In addition, uncoupling protein 1 (UCP1 and UCP2 protein levels, mitochondrial number, and mitochondrial cristae density were upregulated in Vgf-/Vgf- BAT. Using immunohistochemical and histochemical techniques, we detected VGF in nerve fibers innervating BAT and Vgf promoter-driven reporter expression in cervical and thoracic spinal ganglia that project to and innervate the chest wall and tissues including BAT. Moreover, VGF peptide levels were quantified by radioimmunoassay in BAT, and were found to be down-regulated by a high fat diet. Lastly, despite being hypermetabolic

  6. 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.

  7. Creatine transporter (CrT; Slc6a8 knockout mice as a model of human CrT deficiency.

    Directory of Open Access Journals (Sweden)

    Matthew R Skelton

    Full Text Available Mutations in the creatine (Cr transporter (CrT; Slc6a8 gene lead to absence of brain Cr and intellectual disabilities, loss of speech, and behavioral abnormalities. To date, no mouse model of CrT deficiency exists in which to understand and develop treatments for this condition. The purpose of this study was to generate a mouse model of human CrT deficiency. We created mice with exons 2-4 of Slc6a8 flanked by loxP sites and crossed these to Cre:CMV mice to create a line of ubiquitous CrT knockout expressing mice. Mice were tested for learning and memory deficits and assayed for Cr and neurotransmitter levels. Male CrT(⁻/y (affected mice lack Cr in the brain and muscle with significant reductions of Cr in other tissues including heart and testes. CrT(⁻/y mice showed increased path length during acquisition and reversal learning in the Morris water maze. During probe trials, CrT(⁻/y mice showed increased average distance from the platform site. CrT(⁻/y mice showed reduced novel object recognition and conditioned fear memory compared to CrT(+/y. CrT(⁻/y mice had increased serotonin and 5-hydroxyindole acetic acid in the hippocampus and prefrontal cortex. Ubiquitous CrT knockout mice have learning and memory deficits resembling human CrT deficiency and this model should be useful in understanding this disorder.

  8. Analysis of knockout mutants reveals non-redundant functions of poly(ADP-ribose)polymerase isoforms in Arabidopsis.

    Science.gov (United States)

    Pham, Phuong Anh; Wahl, Vanessa; Tohge, Takayuki; de Souza, Laise Rosado; Zhang, Youjun; Do, Phuc Thi; Olas, Justyna J; Stitt, Mark; Araújo, Wagner L; Fernie, Alisdair R

    2015-11-01

    The enzyme poly(ADP-ribose)polymerase (PARP) has a dual function being involved both in the poly(ADP-ribosyl)ation and being a constituent of the NAD(+) salvage pathway. To date most studies, both in plant and non-plant systems, have focused on the signaling role of PARP in poly(ADP-ribosyl)ation rather than any role that can be ascribed to its metabolic function. In order to address this question we here used a combination of expression, transcript and protein localization studies of all three PARP isoforms of Arabidopsis alongside physiological analysis of the corresponding mutants. Our analyses indicated that whilst all isoforms of PARP were localized to the nucleus they are also present in non-nuclear locations with parp1 and parp3 also localised in the cytosol, and parp2 also present in the mitochondria. We next isolated and characterized insertional knockout mutants of all three isoforms confirming a complete knockout in the full length transcript levels of the target genes as well as a reduced total leaf NAD hydrolase activity in the two isoforms (PARP1, PARP2) that are highly expressed in leaves. Physiological evaluation of the mutant lines revealed that they displayed distinctive metabolic and root growth characteristics albeit unaltered leaf morphology under optimal growth conditions. We therefore conclude that the PARP isoforms play non-redundant non-nuclear metabolic roles and that their function is highly important in rapidly growing tissues such as the shoot apical meristem, roots and seeds. PMID:26428915

  9. 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.

  10. Trophic cascades following the disease-induced decline of an apex predator, the Tasmanian devil.

    Science.gov (United States)

    Hollings, Tracey; Jones, Menna; Mooney, Nick; McCallum, Hamish

    2014-02-01

    As apex predators disappear worldwide, there is escalating evidence of their importance in maintaining the integrity and diversity of the ecosystems they inhabit. The largest extant marsupial carnivore, the Tasmanian devil (Sarcophilus harrisii) is threatened with extinction from a transmissible cancer, devil facial tumor disease (DFTD). The disease, first observed in 1996, has led to apparent population declines in excess of 95% in some areas and has spread to more than 80% of their range. We analyzed a long-term Tasmania-wide data set derived from wildlife spotlighting surveys to assess the effects of DFTD-induced devil decline on populations of other mammals and to examine the relative strength of top-down and bottom-up control of mesopredators between 2 regions with different environmental conditions. Collection of the data began >10 years before DFTD was first observed. A decrease in devil populations was immediate across diseased regions following DFTD arrival, and there has been no indication of population recovery. Feral cats (Felis catus) increased in areas where the disease was present the longest, and feral cat occurrence was significantly and negatively associated with devils. The smallest mesopredator, the eastern quoll (Dasyurus viverrinus), declined rapidly following DFTD arrival. This result suggests the species was indirectly protected by devils through the suppression of larger predators. Rainfall deficiency was also a significant predictor of their decline. Environmental variables determined the relative importance of top-down control in the population regulation of mesopredators. In landscapes of low rainfall and relatively higher proportions of agriculture and human settlement, top-down forces were dampened and bottom-up forces had the most effect on mesopredators. For herbivore prey species, there was evidence of population differences after DFTD arrival, but undetected environmental factors had greater effects. The unique opportunity to

  11. Simulation of design basis local events in the APEX-1000 test facility

    International Nuclear Information System (INIS)

    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, onefourth 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. Several design basis events have been simulated at APEX-1000 including the double-ended break of an 8-inch direct vessel injection (DEDVI) line, 2-inch cold leg break, 0.5-inch cold leg break and inadvertent actuation of the automatic depressurization system. Each of these tests show that the core remains covered with ample margin until gravity injection is established regardless of the initiating event. The tests also show that liquid entrainment from the upper plenum which is proportional to the reactor power does not impact the ability of the passive core cooling system to keep the core covered. The results of these tests were used to demonstrate the response of the AP1000 to a spectrum of break sizes and locations, and contributed to the final design approval (FDA) of AP1000. (authors)

  12. 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

  13. 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

  14. 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.

  15. 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.

  16. 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

  17. 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.

  18. 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

  19. 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.

  20. 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.

  1. 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...

  2. 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

  3. 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...

  4. Evaluation of the Scaling of the APEX-1000 Test Facility to AP1000 for Design Basis Events

    International Nuclear Information System (INIS)

    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. Several design basis events have been simulated at APEX-1000 including double-ended direct vessel injection (DEDVI) line break, 2-inch cold leg break, and inadvertent actuation of the automatic depressurization system. These events were analyzed as part of the AP1000 safety analysis using the NOTRUMP computer code. Applying the scaling factors that were used to size the APEX-1000 facility, the test results can be compared to the NOTRUMP plant simulations. These comparisons indicate the similarity between the design basis events in AP1000 and APEX-1000, the adequacy of the scaling of APEX-1000, and the applicability of the NOTRUMP code. (authors)

  5. 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

  6. 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 ...

  7. Environmental enrichment induces behavioural disturbances in neuropeptide Y knockout mice

    Science.gov (United States)

    Reichmann, Florian; Wegerer, Vanessa; Jain, Piyush; Mayerhofer, Raphaela; Hassan, Ahmed M.; Fröhlich, Esther E.; Bock, Elisabeth; Pritz, Elisabeth; Herzog, Herbert; Holzer, Peter; Leitinger, Gerd

    2016-01-01

    Environmental enrichment (EE) refers to the provision of a complex and stimulating housing condition which improves well-being, behaviour and brain function of laboratory animals. The mechanisms behind these beneficial effects of EE are only partially understood. In the current report, we describe a link between EE and neuropeptide Y (NPY), based on findings from NPY knockout (KO) mice exposed to EE. Relative to EE-housed wildtype (WT) animals, NPY KO mice displayed altered behaviour as well as molecular and morphological changes in amygdala and hippocampus. Exposure of WT mice to EE reduced anxiety and decreased central glucocorticoid receptor expression, effects which were absent in NPY KO mice. In addition, NPY deletion altered the preference of EE items, and EE-housed NPY KO mice responded to stress with exaggerated hyperthermia, displayed impaired spatial memory, had higher hippocampal brain-derived neurotrophic factor mRNA levels and altered hippocampal synaptic plasticity, effects which were not seen in WT mice. Accordingly, these findings suggest that NPY contributes to the anxiolytic effect of EE and that NPY deletion reverses the beneficial effects of EE into a negative experience. The NPY system could thus be a target for “enviromimetics”, therapeutics which reproduce the beneficial effects of enhanced environmental stimulation.

  8. Phenotypic characterization of nonsocial behavioral impairment in neurexin 1α knockout rats.

    Science.gov (United States)

    Esclassan, Frederic; Francois, Jennifer; Phillips, Keith G; Loomis, Sally; Gilmour, Gary

    2015-02-01

    Neurexins are neuronal presynaptic proteins that play a key role in mediation of synapse formation. Heterozygous partial deletions in the neurexin-1 gene (NRXN1, 2p16.3) have been observed in autism spectrum disorder (ASD) patients. NRXN1-α knockout (KO) mice present behavioral impairments that resemble some of the core ASD symptoms of social impairment and inflexibility/stereotypy. At present, a thorough assessment of cognitive function has yet to be completed. Rats, containing a biallelic deletion of the NRNX1-α gene on a Sprague Dawley background were compared to littermate wild types across a range of tasks designed to test functional domains disrupted in ASD and other neurodevelopmental disorders, including sensory perception (prepulse inhibition), attention (latent inhibition), associative learning (instrumental and Pavlovian conditioning), and memory (rewarded alternation T maze and spatial discrimination). NRXN1α KO rats were found to present with large and persistent nonsocial deficits, including hyperactivity, deficits in simple instrumental learning, latent inhibition, and spatial-dependent learning. No deficit in sensorimotor gating was observed, despite the presence of an exaggerated startle response. Although KO animals were also able to learn a simple Pavlovian conditioning discrimination, they did display impaired latent inhibition. The presence of pronounced impairments in several domains in NRXN1α KO rats clearly suggests that nonsocial cognitive deficits can also be measured in an animal model of ASD. Further exploration of those deficits, both clinically and preclinically, as planned in the Innovative Medicines Initiative's European Autism Interventions: A Multicenter Study for Developing New Medications program, may help to better understand the brain circuitry involved in ASD and therefore open new avenues to advance novel therapies. PMID:25420124

  9. Acute behavioral effects of nicotine in male and female HINT1 knockout mice.

    Science.gov (United States)

    Jackson, K J; Wang, J B; Barbier, E; Chen, X; Damaj, M I

    2012-11-01

    Human genetic association and brain expression studies, and mouse behavioral and molecular studies implicate a role for the histidine triad nucleotide-binding protein 1 (HINT1) in schizophrenia, bipolar disorder, depression and anxiety. The high comorbidity between smoking and psychiatric disorders, schizophrenia in particular, is well established. Associations with schizophrenia and HINT1 are also sex specific, with effects more predominant in males; however, it is unknown if sex differences associated with the gene extend to other phenotypes. Thus, in this study, using a battery of behavioral tests, we elucidated the role of HINT1 in acute nicotine-mediated behaviors using male and female HINT1 wild-type (+/+) and knockout (-/-) mice. The results show that male HINT1 -/- mice were less sensitive to acute nicotine-induced antinociception in the tail-flick, but not hot-plate test. At low nicotine doses, male and female HINT1 -/- mice were less sensitive to nicotine-induced hypomotility, although the effect was more pronounced in females. Baseline differences in locomotor activity observed in male HINT1 +/+ and -/- mice were absent in females. Nicotine did not produce an anxiolytic effect in male HINT1 -/- mice, but rather an anxiogenic response. Diazepam also failed to induce an anxiolytic response in these mice, suggesting a general anxiety phenotype not specific to nicotine. Differences in anxiety-like behavior were not observed in female mice. These results further support a role for HINT1 in nicotine-mediated behaviors and suggest that alterations in the gene may have differential effects on phenotype in males and females. PMID:22827509

  10. Comprehensive behavioral analysis of pituitary adenylate cyclase-activating polypeptide (PACAP knockout mice

    Directory of Open Access Journals (Sweden)

    Satoko eHattori

    2012-10-01

    Full Text Available Pituitary adenylate cyclase-activating polypeptide (PACAP is a neuropeptide acting as a neurotransmitter, neuromodulator, or neurotrophic factor. PACAP is widely expressed throughout the brain and exerts its functions through the PACAP-specific receptor (PAC1. Recent studies reveal that genetic variants of the PACAP and PAC1 genes are associated with mental disorders, and several behavioral abnormalities of PACAP knockout (KO mice are reported. However, an insufficient number of backcrosses was made using PACAP KO mice on the C57BL/6J background due to their postnatal mortality. To elucidate the effects of PACAP on neuropsychiatric function, the PACAP gene was knocked out in F1 hybrid mice (C57BL/6J x 129SvEv for appropriate control of the genetic background. The PACAP KO mice were then subjected to a behavioral test battery. PACAP deficiency had no significant effects on neurological screen. As shown previously, the mice exhibited significantly increased locomotor activity in a novel environment and abnormal anxiety-like behavior, while no obvious differences between genotypes were shown in home cage activity. In contrast to previous reports, the PACAP KO mice showed normal prepulse inhibition and slightly decreased depression-like behavior. Previous study demonstrates that the social interaction in a resident-intruder test was decreased in PACAP KO mice. On the other hand, we showed that PACAP KO mice exhibited increased social interaction in Crawley’s three-chamber social approach test, although PACAP KO had no significant impact on social interaction in a home cage. PACAP KO mice also exhibited mild performance deficit in working memory in an eight-arm radial maze and the T-maze, while they did not show any significant abnormalities in the left-right discrimination task in the T-maze. These results suggest that PACAP has an important role in the regulation of locomotor activity, social behavior, anxiety-like behavior and, potentially

  11. Functional deficits in nNOSmu-deficient skeletal muscle: myopathy in nNOS knockout mice.

    Directory of Open Access Journals (Sweden)

    Justin M Percival

    Full Text Available Skeletal muscle nNOSmu (neuronal nitric oxide synthase mu localizes to the sarcolemma through interaction with the dystrophin-associated glycoprotein (DAG complex, where it synthesizes nitric oxide (NO. Disruption of the DAG complex occurs in dystrophinopathies and sarcoglycanopathies, two genetically distinct classes of muscular dystrophy characterized by progressive loss of muscle mass, muscle weakness and increased fatigability. DAG complex instability leads to mislocalization and downregulation of nNOSmu; but this is thought to play a minor role in disease pathogenesis. This view persists without knowledge of the role of nNOS in skeletal muscle contractile function in vivo and has influenced gene therapy approaches to dystrophinopathy, the majority of which do not restore sarcolemmal nNOSmu. We address this knowledge gap by evaluating skeletal muscle function in nNOS knockout (KN1 mice using an in situ approach, in which the muscle is maintained in its normal physiological environment. nNOS-deficiency caused reductions in skeletal muscle bulk and maximum tetanic force production in male mice only. Furthermore, nNOS-deficient muscles from both male and female mice exhibited increased susceptibility to contraction-induced fatigue. These data suggest that aberrant nNOSmu signaling can negatively impact three important clinical features of dystrophinopathies and sarcoglycanopathies: maintenance of muscle bulk, force generation and fatigability. Our study suggests that restoration of sarcolemmal nNOSmu expression in dystrophic muscles may be more important than previously appreciated and that it should be a feature of any fully effective gene therapy-based intervention.

  12. Altered social behaviours in neurexin 1α knockout mice resemble core symptoms in neurodevelopmental disorders.

    Directory of Open Access Journals (Sweden)

    Hannah Mary Grayton

    Full Text Available BACKGROUND: Copy number variants have emerged as an important genomic cause of common, complex neurodevelopmental disorders. These usually change copy number of multiple genes, but deletions at 2p16.3, which have been associated with autism, schizophrenia and mental retardation, affect only the neurexin 1 gene, usually the alpha isoform. Previous analyses of neurexin 1α (Nrxn1α knockout (KO mouse as a model of these disorders have revealed impairments in synaptic transmission but failed to reveal defects in social behaviour, one of the core symptoms of autism. METHODS: We performed a detailed investigation of the behavioural effects of Nrxn1α deletion in mice bred onto a pure genetic background (C57BL/6J to gain a better understanding of its role in neurodevelopmental disorders. Wildtype, heterozygote and homozygote Nrxn1α KO male and female mice were tested in a battery of behavioural tests (n = 9-16 per genotype, per sex. RESULTS: In homozygous Nrxn1α KO mice, we observed altered social approach, reduced social investigation, and reduced locomotor activity in novel environments. In addition, male Nrxn1α KO mice demonstrated an increase in aggressive behaviours. CONCLUSIONS: These are the first experimental data that associate a deletion of Nrxn1α with alterations of social behaviour in mice. Since this represents one of the core symptom domains affected in autism spectrum disorders and schizophrenia in humans, our findings suggest that deletions within NRXN1 found in patients may be responsible for the impairments seen in social behaviours, and that the Nrxn1α KO mice are a useful model of human neurodevelopmental disorder.

  13. A selective histone deacetylase-6 inhibitor improves BDNF trafficking in hippocampal neurons from Mecp2 knockout mice:implications for Rett syndrome

    Directory of Open Access Journals (Sweden)

    Xin eXu

    2014-03-01

    Full Text Available Rett syndrome (RTT is a neurodevelopmental disorder caused by loss-of-function mutations in the transcriptional modulator methyl-CpG-binding protein 2 (MECP2. One of the most prominent gene targets of MeCP2 is brain-derived neurotrophic factor (Bdnf, a potent modulator of activity-dependent synaptic development, function and plasticity. Dysfunctional BDNF signaling has been demonstrated in several pathophysiological mechanisms of RTT disease progression. To evaluate whether the dynamics of BDNF trafficking is affected by Mecp2 deletion, we analyzed movements of BDNF tagged with yellow fluorescent protein (YFP in cultured hippocampal neurons by time-lapse fluorescence imaging. We found that both anterograde and retrograde vesicular trafficking of BDNF-YFP are significantly impaired in Mecp2 knockout hippocampal neurons. Selective inhibitors of histone deacetylase 6 (HDAC6 show neuroprotective effects in neurodegenerative diseases and stimulate microtubule-dependent vesicular trafficking of BDNF-containing dense core vesicles. Here, we show that the selective HDAC6 inhibitor Tubastatin-A increased the velocity of BDNF-YFP vesicles in Mecp2 knockout neurons in both directions by increasing αtubulin acetylation. Tubastatin-A also restored activity-dependent BDNF release from Mecp2 knockout neurons to levels comparable to those shown by wildtype neurons. These findings demonstrate that a selective HDAC6 inhibitor is a potential pharmacological strategy to reverse cellular and synaptic impairments in RTT resulting from impaired BDNF signaling.

  14. 锌指核酸酶技术制备肌肉生长抑制素基因敲除的五指山小型猪成纤维细胞%Production of myostatin gene knockout Wuzhishan miniature pig fibroblasts with zinc-finger nucleases

    Institute of Scientific and Technical Information of China (English)

    曹随忠; 岳成鹤; 李西睿; 冯冲; 龙川; 潘登科

    2013-01-01

    Disruption of myostatin (MSTN) gene in pigs may improve porcine lean meat percentage (LMP), and create an animal model for certain human diseases. Using zinc-finger nucleases (ZFNs) technology, MSTN gene was deleted in Wuzhishan miniature pig fibroblasts by transfection of either ZFNs plasmids or ZFNs mRNA in high efficiency. Strikingly, ZFNs encoding mRNA could produce MSTN+/-and MSTN-/- cell colonies with single or double allele deletion by single transfection. Sequencing results demonstrated that 92.18% of the mutations were short fragment deletions or insertions (≤10 bp). Prediction of amino acids sequences indicated that more than half of the mutations cause premature transla-tional-termination codon. MSTN+/+, MSTN+/-, and MSTK-/- cell colonies were used as nuclear donor for somatic cell nuclear transfer (SCNT), and developmental potential of SCNT embryos were measured by the blastocyst rate. The results revealed no significant difference in development competence among the three kinds of reconstructed embryos (14.29% vs. 19.64% vs. 16.13%), which provides the possibility of making myostatin knock out pigs in the future.%敲除猪肌肉生长抑制素(Myostatin,MSTN)基因可能提高猪瘦肉率,MSTN 基因敲除猪也可作为相关疾病的动物模型.文章利用锌指核酸酶(Zinc-finger nucleases,ZFNs)技术敲除五指山小型猪胎儿成纤维细胞MSTN基因,为制备MSTN 基因敲除猪奠定基础.ZFNs 质粒或编码ZFNs 的mRNA 均能高效敲除MSTN 基因,使用ZFNs mRNA 能直接得到MSTN+/-和MSTN-/-两种基因型的细胞克隆.DNA 序列测定与分析发现,细胞克隆的突变类型多为ZFNs 作用靶位点处不大于10 bp 的碱基插入或缺失(92.18 %); 氨基酸预测发现,突变型MSTN 基因的终止密码子常常提前出现.将MSTN 基因敲除的细胞进行体细胞核移植(Somatic cell nuclear transfer,SCNT)发现,胚胎体外早期发育潜力与野生型无显著差异,表明这些细胞可用于后续MSTN 基因敲除猪的制备.

  15. Hypoxic and hypercapnic challenges unveil respiratory vulnerability of Surf1 knockout mice, an animal model of Leigh syndrome.

    Science.gov (United States)

    Stettner, Georg M; Viscomi, Carlo; Zeviani, Massimo; Wilichowski, Ekkehard; Dutschmann, Mathias

    2011-05-01

    Surf1 gene mutations were detected as a main cause for Leigh syndrome (LS), also known as infantile subacute necrotizing encephalomyelopathy. This syndrome which is commonly associated with systemic cytochrome c oxidase (COX) deficiency manifests in early childhood and has an invariable poor prognosis. Progressive disturbances of the respiratory function, for which both the metabolic condition and necrotizing brainstem lesions contribute, belong to the major symptoms of LS. A constitutive knockout (KO) mouse for Surf1 enables invasive investigations of distinct aspects of LS. In the present study the respiratory function was analyzed applying an arterially perfused brainstem preparation. Compared to wild type (WT) preparations Surf1 KO preparations had a higher baseline respiratory frequency and abnormal responses to hypoxia and hypercapnia that involved both respiratory frequency and motor nerve discharge pattern. These data suggest that COX deficiency impairs peripheral and/or central chemoreceptor function. PMID:21167962

  16. Brain response to traumatic brain injury in wild-type and interleukin-6 knockout mice: a microarray analysis

    DEFF Research Database (Denmark)

    Poulsen, Christian Bjørn; Penkowa, Milena; Borup, Rehannah; Nielsen, Finn Cilius; Cáceres, Mario; Quintana, Albert; Molinero, Amalia; Carrasco, Javier; Giralt, Mercedes; Hidalgo, Juan

    2005-01-01

    Traumatic injury to the brain is one of the leading causes of injury-related death or disability. Brain response to injury is orchestrated by cytokines, such as interleukin (IL)-6, but the full repertoire of responses involved is not well known. We here report the results obtained with microarrays...... processes involved in the initial tissue injury and later regeneration of the parenchyma. IL-6 deficiency showed a dramatic effect in the expression of many genes, especially in the 1 day post-lesion timing, which presumably underlies the poor capacity of IL-6 knockout mice to cope with brain damage. The...... results highlight the importance of IL-6 controlling the response of the brain to injury as well as the suitability of microarrays for identifying specific targets worthy of further study....

  17. [Expression analysis of green fluorescent protein in tissues and organs in α-1,3 galactosyltransferase knockout pigs].

    Science.gov (United States)

    Zhifang, Li; Chong, Feng; Huili, Ji; Ningning, Shi; Xiaofeng, Song; Qinli, Zhao; Chuan, Long; Dengke, Pan; Xiaogan, Yang

    2015-12-01

    The pig is an ideal source to provide organs because its organ size and physiology are similar to humans. However, an acute rejection will ensue after pig-to-human xenotransplantation. The α-1,3 galactosyltransferase gene knockout (GTKO) pigs were generated in recent years, and could solve the problem of hyperacute rejection. But due to lack of reporting genes, the rejection status of cells and organs post pig-to-human xenotransplantation cannot be visualized. In this study, we introduced the enhanced green fluorescent protein (EGFP) gene driven by the CAG promoter into GTKO porcine ear fibroblasts. Then we produced transgenic pigs expressing the EGFP gene by nuclear transfer technology. Expression levels of EGFP in different tissues and organs of the cloned pig were investigated by Nightsea DFP-1 Fluorescent Protein Flashlight, fluorescence microscope and quantitative PCR assays. The results showed that the protein and transcript of EGFP were expressed in all tissues and organs of the GTKO pig, but the expression was weak in the liver and central nervous system. In conclusion, we have successfully produced the transgenic GTKO pigs expressing EGFP in all tested tissues and organs, which builds up a good basis to track transplanted cells or tissues. PMID:26704946

  18. Targeted Heritable Mutation and Gene Conversion by Cas9-CRISPR in Caenorhabditis elegans

    OpenAIRE

    Katic, Iskra; Großhans, Helge

    2013-01-01

    We have achieved targeted heritable genome modification in Caenorhabditis elegans by injecting mRNA of the nuclease Cas9 and Cas9 guide RNAs. This system rapidly creates precise genomic changes, including knockouts and transgene-instructed gene conversion.

  19. Improved cognitive flexibility in serotonin transporter knockout rats is unchanged following chronic cocaine self-administration

    NARCIS (Netherlands)

    Nonkes, L.J.; Maes, J.H.R.; Homberg, J.R.

    2013-01-01

    Cocaine dependence is associated with orbitofrontal cortex (OFC)-dependent cognitive inflexibility in both humans and laboratory animals. A critical question is whether cocaine self-administration affects pre-existing individual differences in cognitive flexibility. Serotonin transporter knockout (5

  20. 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

  1. 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

  2. 15q13.3 homozygous knockout mouse model display epilepsy-, autism- and schizophrenia-related phenotypes.

    Science.gov (United States)

    Forsingdal, A; Fejgin, K; Nielsen, V; Werge, T; Nielsen, J

    2016-01-01

    The 15q13.3 microdeletion syndrome is caused by a 1.5-MB hemizygous microdeletion located on 15q13.3 affecting seven genes: FAN1; MTMR10; TRPM1; miR-211; KLF13; OTUD7A; and CHRNA7. The 15q13.3 microdeletion increases the risk of intellectual disability, epilepsy, autism spectrum disorder and schizophrenia, though the clinical profile varies considerably. Two mouse models of this syndrome, with hemizygous deletion of the orthologous region in the murine genome, have recently been shown to recapitulate a number of the behavioral and physiological deficits that characterize the human condition. Still, little is known of the underlying biological mechanisms. Eleven human cases with homozygous deletion of the 15q13.3 region have been reported, all with severe functional and physiological impairments. We therefore hypothesized that a 15q13.3 homozygous knockout would confer more pronounced behavioral and physiological deficits in mice than the 15q13.3 hemizygous deletion. Here we report the characterization of a 15q13.3 knockout mouse. We observed marked deficits including altered seizure susceptibility, autistic behavior-related phenotypes, and auditory sensory processing. Several of these deficits, albeit less pronounced, were also found in the 15q13.3 hemizygous littermates indicating a gene-dosage dependency. Our findings strongly indicate that studies of the hemi- and homozygous 15q13.3 mouse strains will facilitate understanding of the biological mechanisms of severe mental disorders. PMID:27459725

  3. Maximal Oxygen Consumption Is Reduced in Aquaporin-1 Knockout Mice.

    Science.gov (United States)

    Al-Samir, Samer; Goossens, Dominique; Cartron, Jean-Pierre; Nielsen, Søren; Scherbarth, Frank; Steinlechner, Stephan; Gros, Gerolf; Endeward, Volker

    2016-01-01

    We have measured maximal oxygen consumption ([Formula: see text]O2,max) of mice lacking one or two of the established mouse red-cell CO2 channels AQP1, AQP9, and Rhag. We intended to study whether these proteins, by acting as channels for O2, determine O2 exchange in the lung and in the periphery. We found that [Formula: see text]O2,max as determined by the Helox technique is reduced by ~16%, when AQP1 is knocked out, but not when AQP9 or Rhag are lacking. This figure holds for animals respiring normoxic as well as hypoxic gas mixtures. To see whether the reduction of [Formula: see text]O2,max is due to impaired O2 uptake in the lung, we measured carotid arterial O2 saturation (SO2) by pulse oximetry. Neither under normoxic (inspiratory O2 21%) nor under hypoxic conditions (11% O2) is there a difference in SO2 between AQP1null and WT mice, suggesting that AQP1 is not critical for O2 uptake in the lung. The fact that the % reduction of [Formula: see text]O2,max is identical in normoxia and hypoxia indicates moreover that the limitation of [Formula: see text]O2,max is not due to an O2 diffusion problem, neither in the lung nor in the periphery. Instead, it appears likely that AQP1null animals exhibit a reduced [Formula: see text]O2,max due to the reduced wall thickness and muscle mass of the left ventricles of their hearts, as reported previously. We conclude that very likely the properties of the hearts of AQP1 knockout mice cause a reduced maximal cardiac output and thus cause a reduced [Formula: see text]O2,max, which constitutes a new phenotype of these mice. PMID:27559317

  4. 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

  5. Altered sleep homeostasis in rev-erbα knockout mice

    OpenAIRE

    Mang, Géraldine M.; La Spada, Francesco; Emmenegger, Yann; Chappuis, Sylvie; Ripperger, Jürgen A; Albrecht, Urs; Franken, Paul

    2016-01-01

    Study Objectives: The nuclear receptor REV-ERBα is a potent, constitutive transcriptional repressor critical for the regulation of key circadian and metabolic genes. Recently, REV-ERBα's involvement in learning, neurogenesis, mood, and dopamine turnover was demonstrated suggesting a specific role in central nervous system functioning. We have previously shown that the brain expression of several core clock genes, including Rev-erbα, is modulated by sleep loss. We here test the consequen...

  6. 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...

  7. Oroclinal bending and orogenic asymmetry? A case study from the apex of the Northern Andean orocline

    Science.gov (United States)

    Mora, A.; Parra-Amezquita, M.; Rodriguez, G.

    2013-05-01

    Here we present new thermocronometric analysis from the eastern flank of the Colombian Eastern Cordillera at 7° N lat. which aid in deconvolving the roles of plate tectonics, tectonic inheritance and surface processes in building the Cocuy syntaxis, at the apex of the Northern Andean orocline. The Cocuy syntaxis is the region with higher structural and topographic relief in the Eastern Cordillera. The primary factor controlling that is faster tectonism, apparently related with the most important Panama collision at ca. 4 Ma. This push from behind is focused between two resistant plates and escapes toward a weak foreland plate, which is able to flex. However, we suggest it is an enhanced late Miocene to Recent exhumation episode which is responsible for the limited advance of the deformation front as basement involved blocks. Faster denudation causes faster sedimentation rates in the weak foreland plate east of Cocuy. In this case, the thick pile of synkinematic sediments would make an otherwise possible thin skin deformation migration very difficult.

  8. 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...

  9. 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.

  10. 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

  11. 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.

  12. Inhibition of white light of 86Rb+ absorption in the root apex of corn

    International Nuclear Information System (INIS)

    Measurements of cell lengths made at 0.5 millimeter intervals in median longitudinal sections of the primary roots of corn (Zea mays) were used to construct a growth curve. The region 1.5 to 4.0 millimeters from the apex contained the largest number of elongating cells. Absorption of 86Rb+ was measured using intact, dark-grown corn seedlings. Following uptake and exchange, the terminal 8.0 millimeters of each root was cut into four 2.0 millimeter segments. Maximum 86Rb+ uptake occurred in the region from 0.0 to 4.0 millimeter from the root tip. Washing the intact primary root in fresh 2.0 millimolar CaSO4 for 2 hours prior to uptake augmented the rate of 86Rb+ uptake in all regions. Illumination with white light during washing caused a reduction of 86Rb+ uptake as compared with controls washing in darkness, and the region of greatest light response was the region of elongation. Removal of the coleoptile prior to washing did not prevent the light inhibition of subsequent 86Rb+ uptake. Removal of the root cap prior to washing in light partially reversed the light-induced inhibition of the washing response

  13. 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.

  14. 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.

  15. 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. PMID:27123930

  16. Atp6v0a4 knockout mouse is a model of distal renal tubular acidosis with hearing loss, with additional extrarenal phenotype

    OpenAIRE

    Elizabeth E. Norgett; Golder, Zoe J.; Lorente-Cánovas, Beatriz; Ingham, Neil; Steel, Karen P; Karet Frankl, Fiona E.

    2012-01-01

    Autosomal recessive distal renal tubular acidosis (dRTA) is a severe disorder of acid–base homeostasis, often accompanied by sensorineural deafness. We and others have previously shown that mutations in the tissue-restricted a4 and B1 subunits of the H+-ATPase underlie this syndrome. Here, we describe an Atp6v0a4 knockout mouse, which lacks the a4 subunit. Using β-galactosidase as a reporter for the null gene, developmental a4 expression was detected in developing bone, nose, eye, and skin, i...

  17. Knockout of Na-glucose transporter SGLT2 attenuates hyperglycemia and glomerular hyperfiltration but not kidney growth or injury in diabetes mellitus

    OpenAIRE

    Vallon, Volker; Rose, Michael; Gerasimova, Maria; Satriano, Joseph; Platt, Kenneth A.; Koepsell, Hermann; Cunard, Robyn; Sharma, Kumar; Thomson, Scott C.; Rieg, Timo

    2012-01-01

    The Na-glucose cotransporter SGLT2 mediates high-capacity glucose uptake in the early proximal tubule and SGLT2 inhibitors are developed as new antidiabetic drugs. We used gene-targeted Sglt2 knockout (Sglt2−/−) mice to elucidate the contribution of SGLT2 to blood glucose control, glomerular hyperfiltration, kidney growth, and markers of renal growth and injury at 5 wk and 4.5 mo after induction of low-dose streptozotocin (STZ) diabetes. The absence of SGLT2 did not affect renal mRNA expressi...

  18. Msx homeobox gene family and craniofacial development

    Institute of Scientific and Technical Information of China (English)

    SYLVIA ALAPPAT; ZUN YI ZHANG; YI PING CHEN

    2003-01-01

    Vertebrate Msx genes are unlinked,homeobox-containing genes that bear homology to the Drosophila muscle segment homeobox gene.These genes are expressed at multiple sites of tissue-tissue interactions during vertebrate embryonic development.Inductive interactions mediated by the Msx genes are essential for normal craniofacial,limb and ectodermal organ morphogenesis,and are also essential to survival in mice,as manifested by the phenotypic abnormalities shown in knockout mice and in humans.This review summarizes studies on the expression,regulation,and functional analysis of Msx genes that bear relevance to craniofacial development in humans and mice.

  19. 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.

  20. 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...

  1. 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...

  2. 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...

  3. 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...

  4. Contemporary methods for determining root canal working length in primary teeth: digital radiography and electronic apex locators

    OpenAIRE

    Zeren, Ali Emre; Sarı, Şaziye

    2014-01-01

    With developing technology, in endodontics, like in other fields of dentistry, different approaches are introduced as means for improving success, efficacy and comfort of treatment. In pedodontics, a branch of dentistry where patient compliance and treatment periods are crucial, the new treatment approaches help dentists perform more efficient and faster. Currently, digital radiography and apex locators are widely used in the root canal treatment of permanent teeth. The aim of this review was...

  5. Left ventricular diverticulum with marked hypertrophy of the left ventricular apex revealed by thallium-201 myocardial emission CT

    International Nuclear Information System (INIS)

    A case of left ventricular apical diverticulum with marked hypertrophy of the left ventricular apical wall revealed by thallium-201 myocardial emission CT is reported. A 23-year-old woman was admitted to our hospital for evaluation of chest oppression. She was known to have had a heart murmur soon after birth, but she grew uneventfully, partaking in normal exercise. At the age of 21, she began to feel chest oppression during exercise. As the attacks became frequent, she was admitted to our hospital. Physical examination revealed an ejection systolic murmur in the second left intercostal space. Electrocardiography showed ST depression and T inversion in leads III, a VF and V4-6. M-mode echocardiography was normal. Two-dimensional echocardiography showed a small diverticulum at the apex of the left ventricle, which was also recognized by left ventriculography. It was about 8 x 12 mm in size. Thallium-201 myocardial emission CT disclosed marked uptake in the apex of the left ventricle, suggesting apical hypertrophy. Stress thallium-201 myocardial emission CT was negative. Coronary angiography was normal. The cause of chest oppression in this patient is uncertain, but the small diverticulum and hypertrophy of the cardiac apex may play a role in its pathogenesis. (author)

  6. Autism spectrum disorder traits in Slc9a9 knock-out mice.

    Science.gov (United States)

    Yang, Lina; Faraone, Stephen V; Zhang-James, Yanli

    2016-04-01

    Autism spectrum disorders (ASDs) are a group of neurodevelopmental disorders which begin in childhood and persist into adulthood. They cause lifelong impairments and are associated with substantial burdens to patients, families, and society. Genetic studies have implicated the sodium/proton exchanger (NHE) nine gene, Slc9a9, to ASDs and attention-deficit/hyperactivity disorder(ADHD). Slc9a9 encodes, NHE9, a membrane protein of the late recycling endosomes. The recycling endosome plays an important role in synapse development and plasticity by regulating the trafficking of membrane neurotransmitter receptors and transporters. Here we tested the hypothesis that Slc9a9 knock-out (KO) mice would show ADHD-like and ASD-like traits. Ultrasonic vocalization (USV) recording showed that Slc9a9 KO mice emitted fewer calls and had shorter call durations, which suggest communication impairment. Slc9a9 KO mice lacked a preference for social novelty, but did not show deficits in social approach; Slc9a9 KO mice spent more time self-grooming, an indicator for restricted and repetitive behavior. We did not observe hyperactivity or other behavior impairments which are commonly comorbid with ASDs in human, such as anxiety-like behavior. Our study is the first animal behavior study that links Slc9a9 to ASDs. By eliminatingNHE9 activity, it provides strong evidence that lack of Slc9a9leads to ASD-like behaviors in mice and provides the field with a new mouse model of ASDs. PMID:26755066

  7. Blue-Green Algae Inhibit the Development of Atherosclerotic Lesions in Apolipoprotein E Knockout Mice.

    Science.gov (United States)

    Ku, Chai Siah; Kim, Bohkyung; Pham, Tho X; Yang, Yue; Wegner, Casey J; Park, Young-Ki; Balunas, Marcy; Lee, Ji-Young

    2015-12-01

    Hyperlipidemia and inflammation contribute to the development of atherosclerotic lesions. Our objective was to determine antiatherogenic effect of edible blue-green algae (BGA) species, that is, Nostoc commune var. sphaeroides Kützing (NO) and Spirulina platensis (SP), in apolipoprotein E knockout (ApoE(-/-)) mice, a well-established mouse model of atherosclerosis. Male ApoE(-/-) mice were fed a high-fat/high-cholesterol (HF/HC, 15% fat and 0.2% cholesterol by wt) control diet or a HF/HC diet supplemented with 5% (w/w) of NO or SP powder for 12 weeks. Plasma total cholesterol (TC) and triglycerides (TG) were measured, and livers were analyzed for histology and gene expression. Morphometric analysis for lesions and immunohistochemical analysis for CD68 were conducted in the aorta and the aortic root. NO supplementation significantly decreased plasma TC and TG, and liver TC, compared to control and SP groups. In the livers of NO-fed mice, less lipid droplets were present with a concomitant decrease in fatty acid synthase protein levels than the other groups. There was a significant increase in hepatic low-density lipoprotein receptor protein levels in SP-supplemented mice than in control and NO groups. Quantification of aortic lesions by en face analysis demonstrated that both NO and SP decreased aortic lesion development to a similar degree compared with control. While lesions in the aortic root were not significantly different between groups, the CD68-stained area in the aortic root was significantly lowered in BGA-fed mice than controls. In conclusion, both NO and SP supplementation decreased the development of atherosclerotic lesions, suggesting that they may be used as a natural product for atheroprotection. PMID:26566121

  8. Somatostatin receptor 1 and 5 double knockout mice mimic neurochemical changes of Huntington's disease transgenic mice.

    Directory of Open Access Journals (Sweden)

    Padmesh S Rajput

    Full Text Available BACKGROUND: Selective degeneration of medium spiny neurons and preservation of medium sized aspiny interneurons in striatum has been implicated in excitotoxicity and pathophysiology of Huntington's disease (HD. However, the molecular mechanism for the selective sparing of medium sized aspiny neurons and vulnerability of projection neurons is still elusive. The pathological characteristic of HD is an extensive reduction of the striatal mass, affecting caudate putamen. Somatostatin (SST positive neurons are selectively spared in HD and Quinolinic acid/N-methyl-D-aspartic acid induced excitotoxicity, mimic the model of HD. SST plays neuroprotective role in excitotoxicity and the biological effects of SST are mediated by five somatostatin receptor subtypes (SSTR1-5. METHODS AND FINDINGS: To delineate subtype selective biological responses we have here investigated changes in SSTR1 and 5 double knockout mice brain and compared with HD transgenic mouse model (R6/2. Our study revealed significant loss of dopamine and cAMP regulated phosphoprotein of 32 kDa (DARPP-32 and comparable changes in SST, N-methyl-D-aspartic acid receptors subtypes, calbindin and brain nitric oxide synthase expression as well as in key signaling proteins including calpain, phospho-extracellular-signal-regulated kinases1/2, synapsin-IIa, protein kinase C-α and calcineurin in SSTR1/5(-/- and R6/2 mice. Conversely, the expression of somatostatin receptor subtypes, enkephalin and phosphatidylinositol 3-kinases were strain specific. SSTR1/5 appears to be important in regulating NMDARs, DARPP-32 and signaling molecules in similar fashion as seen in HD transgenic mice. CONCLUSIONS: This is the first comprehensive description of disease related changes upon ablation of G- protein coupled receptor gene. Our results indicate that SST and SSTRs might play an important role in regulation of neurodegeneration and targeting this pathway can provide a novel insight in understanding the

  9. Experimental evidence for the involvement of PDLIM5 in mood disorders in hetero knockout mice.

    Directory of Open Access Journals (Sweden)

    Yasue Horiuchi

    Full Text Available BACKGROUND: Reports indicate that PDLIM5 is involved in mood disorders. The PDLIM5 (PDZ and LIM domain 5 gene has been genetically associated with mood disorders; it's expression is upregulated in the postmortem brains of patients with bipolar disorder and downregulated in the peripheral lymphocytes of patients with major depression. Acute and chronic methamphetamine (METH administration may model mania and the evolution of mania into psychotic mania or schizophrenia-like behavioral changes, respectively. METHODS: To address whether the downregulation of PDLIM5 protects against manic symptoms and cause susceptibility to depressive symptoms, we evaluated the effects of reduced Pdlim5 levels on acute and chronic METH-induced locomotor hyperactivity, prepulse inhibition, and forced swimming by using Pdlim5 hetero knockout (KO mice. RESULTS: The homozygous KO of Pdlim5 is embryonic lethal. The effects of METH administration on locomotor hyperactivity and the impairment of prepulse inhibition were lower in Pdlim5 hetero KO mice than in wild-type mice. The transient inhibition of PDLIM5 (achieved by blocking the translocation of protein kinase C epsilon before the METH challenge had a similar effect on behavior. Pdlim5 hetero KO mice showed increased immobility time in the forced swimming test, which was diminished after the chronic administration of imipramine. Chronic METH treatment increased, whereas chronic haloperidol treatment decreased, Pdlim5 mRNA levels in the prefrontal cortex. Imipramine increased Pdlim5 mRNA levels in the hippocampus. CONCLUSION: These findings are partially compatible with reported observations in humans, indicating that PDLIM5 is involved in psychiatric disorders, including mood disorders.

  10. Brain-derived neurotrophic factor signaling is altered in the forebrain of Engrailed-2 knockout mice.

    Science.gov (United States)

    Zunino, G; Messina, A; Sgadò, P; Baj, G; Casarosa, S; Bozzi, Y

    2016-06-01

    Engrailed-2 (En2), a homeodomain transcription factor involved in regionalization and patterning of the midbrain and hindbrain regions has been associated to autism spectrum disorders (ASDs). En2 knockout (En2(-/-)) mice show ASD-like features accompanied by a significant loss of GABAergic subpopulations in the hippocampus and neocortex. Brain-derived neurotrophic factor (BDNF) is a crucial factor for the postnatal development of forebrain GABAergic neurons, and altered GABA signaling has been hypothesized to underlie the symptoms of ASD. Here we sought to determine whether interneuron loss in the En2(-/-) forebrain might be related to altered expression of BDNF and its signaling receptors. We first evaluated the expression of different BDNF mRNA isoforms in the neocortex and hippocampus of wild-type (WT) and En2(-/-) mice. Quantitative RT-PCR showed a marked down-regulation of several splicing variants of BDNF mRNA in the neocortex but not hippocampus of adult En2(-/-) mice, as compared to WT controls. Accordingly, levels of mature BDNF protein were lower in the neocortex but not hippocampus of En2(-/-) mice, as compared to WT. Increased levels of phosphorylated TrkB and decreased levels of p75 receptor were also detected in the neocortex of mutant mice. Accordingly, the expression of low density lipoprotein receptor (LDLR) and RhoA, two genes regulated via p75 was significantly altered in forebrain areas of mutant mice. These data indicate that BDNF signaling alterations might be involved in the anatomical changes observed in the En2(-/-) forebrain and suggest a pathogenic role of altered BDNF signaling in this mouse model of ASD. PMID:26987954

  11. Alpha-asarone improves striatal cholinergic function and locomotor hyperactivity in Fmr1 knockout mice.

    Science.gov (United States)

    Qiu, Guozhen; Chen, Shengqiang; Guo, Jialing; Wu, Jie; Yi, Yong-Hong

    2016-10-01

    Hyperactivity is a symptom found in several neurological and psychiatric disorders, including Fragile X syndrome (FXS). The animal model of FXS, fragile X mental retardation gene (Fmr1) knockout (KO) mouse, exhibits robust locomotor hyperactivity. Alpha (α)-asarone, a major bioactive component isolated from Acorus gramineus, has been shown in previous studies to improve various disease conditions including central nervous system disorders. In this study, we show that treatment with α-asarone alleviates locomotor hyperactivity in Fmr1 KO mice. To elucidate the mechanism underlying this improvement, we evaluated the expressions of various cholinergic markers, as well as acetylcholinesterase (AChE) activity and acetylcholine (ACh) levels, in the striatum of Fmr1 KO mice. We also analyzed the AChE-inhibitory activity of α-asarone. Striatal samples from Fmr1 KO mice showed decreased m1 muscarinic acetylcholine receptor (m1 mAChR) expression, increased AChE activity, and reduced ACh levels. Treatment with α-asarone improved m1 mAChR expression and ACh levels, and attenuated the increased AChE activity. In addition, α-asarone dose-dependently inhibited AChE activity in vitro. These results indicate that direct inhibition of AChE activity and up-regulation of m1 mAChR expression in the striatum might contribute to the beneficial effects of α-asarone on locomotor hyperactivity in Fmr1 KO mice. These findings might improve understanding of the neurobiological mechanisms responsible for locomotor hyperactivity. PMID:27316341

  12. CDKL5 knockout leads to altered inhibitory transmission in the cerebellum of adult mice.

    Science.gov (United States)

    Sivilia, S; Mangano, C; Beggiato, S; Giuliani, A; Torricella, R; Baldassarro, V A; Fernandez, M; Lorenzini, L; Giardino, L; Borelli, A C; Ferraro, L; Calzà, L

    2016-06-01

    Mutations in the X-linked cyclin-dependent kinase-like 5 gene (CDKL5) are associated to severe neurodevelopmental alterations including motor symptoms. In order to elucidate the neurobiological substrate of motor symptoms in CDKL5 syndrome, we investigated the motor function, GABA and glutamate pathways in the cerebellum of CDKL5 knockout female mice. Behavioural data indicate that CDKL5-KO mice displayed impaired motor coordination on the Rotarod test, and altered steps, as measured by the gait analysis using the CatWalk test. A higher reduction in spontaneous GABA efflux, than that in glutamate, was observed in CDKL5-KO mouse cerebellar synaptosomes, leading to a significant increase of spontaneous glutamate/GABA efflux ratio in these animals. On the contrary, there were no differences between groups in K(+) -evoked GABA and glutamate efflux. The anatomical analysis of cerebellar excitatory and inhibitory pathways showed a selective defect of the GABA-related marker GAD67 in the molecular layer in CDKL5-KO mice, while the glutamatergic marker VGLUT1 was unchanged in the same area. Fine cerebellar structural abnormalities such as a reduction of the inhibitory basket 'net' estimated volume and an increase of the pinceau estimated volume were also observed in CDKL5-KO mice. Finally, the BDNF mRNA expression level in the cerebellum, but not in the hippocampus, was reduced compared with WT animals. These data suggest that CDKL5 deletion during development more markedly impairs the establishment of a correct GABAergic cerebellar network than that of glutamatergic one, leading to the behavioural symptoms associated with CDKL5 mutation. PMID:27108663

  13. Subchronic exposure to ethyl tertiary butyl ether resulting in genetic damage in Aldh2 knockout mice.

    Science.gov (United States)

    Weng, Zuquan; Suda, Megumi; Ohtani, Katsumi; Mei, Nan; Kawamoto, Toshihiro; Nakajima, Tamie; Wang, Rui-Sheng

    2013-09-15

    Ethyl tertiary butyl ether (ETBE) is biofuel additive recently used in Japan and some other countries. Limited evidence shows that ETBE has low toxicity. Acetaldehyde (AA), however, as one primary metabolite of ETBE, is clearly genotoxic and has been considered to be a potential carcinogen. The aim of this study was to evaluate the effects of ALDH2 gene on ETBE-induced genotoxicity and metabolism of its metabolites after inhalation exposure to ETBE. A group of wild-type (WT) and Aldh2 knockout (KO) C57BL/6 mice were exposed to 500ppm ETBE for 1-6h, and the blood concentrations of ETBE metabolites, including AA, tert-butyl alcohol and 2-methyl-1,2-propanediol, were measured. Another group of mice of WT and KO were exposed to 0, 500, 1750, or 5000ppm ETBE for 6h/day with 5 days per weeks for 13 weeks. Genotoxic effects of ETBE in these mice were measured by the alkaline comet assay, 8-hydroxyguanine DNA-glycosylase modified comet assay and micronucleus test. With short-term exposure to ETBE, the blood concentrations of all the three metabolites in KO mice were significantly higher than the corresponding concentrations of those in WT mice of both sexes. After subchronic exposure to ETBE, there was significant increase in DNA damage in a dose-dependent manner in KO male mice, while only 5000ppm exposure significantly increased DNA damage in male WT mice. Overall, there was a significant sex difference in genetic damage in both genetic types of mice. These results showed that ALDH2 is involved in the detoxification of ETBE and lack of enzyme activity may greatly increase the sensitivity to the genotoxic effects of ETBE, and male mice were more sensitive than females. PMID:23810710

  14. Repetitive grooming and sensorimotor abnormalities in an ephrin-A knockout model for Autism Spectrum Disorders

    Science.gov (United States)

    Wurzman, Rachel; Forcelli, Patrick A.; Griffey, Christopher J.; Kromer, Lawrence F.

    2014-01-01

    EphA receptors and ephrin-A ligands play important roles in neural development and synaptic plasticity in brain regions where expression persists into adulthood. Recently, EPHA3 and EPHA7 gene mutations were linked with Autism Spectrum Disorders (ASDs) and developmental neurological delays, respectively. Furthermore, deletions of ephrin-A2 or ephrin-A3, which exhibit high binding affinity for EphA3 and EphA7 receptors, are associated with subtle deficits in learning and memory behavior and abnormalities in dendritic spine morphology in the cortex and hippocampus in mice. To better characterize a potential role for these ligands in ASDs, we performed a comprehensive behavioral characterization of anxiety-like, sensorimotor, learning, and social behaviors in ephrin-A2/-A3 double knockout (DKO) mice. The predominant phenotype in DKO mice was repetitive and self-injurious grooming behaviors such as have been associated with corticostriatal circuit abnormalities in other rodent models of neuropsychiatric disorders. Consistent with ASDs specifically, DKO mice exhibited decreased preference for social interaction in the social approach assay, decreased locomotor activity in the open field, increased prepulse inhibition of acoustic startle, and a shift towards self-directed activity (e.g., grooming) in novel environments, such as marble burying. Although there were no gross deficits in cognitive assays, subtle differences in performance on fear conditioning and in the Morris water maze resembled traits observed in other rodent models of ASD. We therefore conclude that ephrin-A2/-A3 DKO mice have utility as a novel ASD model with an emphasis on sensory abnormalities and restricted, repetitive behavioral symptoms. PMID:25281279

  15. What Have We Learned from Glycosyltransferase Knockouts in Mice?

    Science.gov (United States)

    Stanley, Pamela

    2016-08-14

    There are five major classes of glycan including N- and O-glycans, glycosaminoglycans, glycosphingolipids, and glycophosphatidylinositol anchors, all expressed at the molecular frontier of each mammalian cell. Numerous biological consequences of altering the expression of mammalian glycans are understood at a mechanistic level, but many more remain to be characterized. Mouse mutants with deleted, defective, or misexpressed genes that encode activities necessary for glycosylation have led the way to identifying key functions of glycans in biology. However, with the advent of exome sequencing, humans with mutations in genes involved in glycosylation are also revealing specific requirements for glycans in mammalian development. The aim of this review is to summarize glycosylation genes that are necessary for mouse embryonic development, pathway-specific glycosylation genes whose deletion leads to postnatal morbidity, and glycosylation genes for which effects are mild, but perturbation of the organism may reveal functional consequences. General strategies for generating and interpreting the phenotype of mice with glycosylation defects are discussed in relation to human congenital disorders of glycosylation (CDG). PMID:27040397

  16. Impaired social behavior in 5-HT3A receptor knockout mice

    Directory of Open Access Journals (Sweden)

    Laura A Smit-Rigter

    2010-11-01

    Full Text Available The 5-HT3 receptor is a ligand-gated ion channel expressed on interneurons throughout the brain. So far, analysis of the 5-HT3A knockout mouse revealed changes in nociceptive processing and a reduction in anxiety related behavior. Recently, it was shown that the 5-HT3 receptor is also expressed on Cajal-Retzius cells which play a key role in cortical development and that knockout mice lacking this receptor showed aberrant growth of the dendritic tree of cortical layer II/III pyramidal neurons. Other mouse models in which serotonergic signaling was disrupted during development showed similar morphological changes in the cortex, and in addition, also deficits in social behavior. Here, we subjected male and female 5-HT3A knockout mice and their non-transgenic littermates to several tests of social behavior. We found that 5-HT3A knockout mice display impaired social communication in the social transmission of food preference task. Interestingly, we showed that in the social interaction test only female 5-HT3A knockout mice spent less time in reciprocal social interaction starting after 5 minutes of testing. Moreover, we observed differences in preference for social novelty for male and female 5-HT3A knockout mice during the social approach test. However, no changes in olfaction, exploratory activity and anxiety were detected. These results indicate that the 5-HT3A knockout mouse displays impaired social behavior with specific changes in males and females, reminiscent to other mouse models in which serotonergic signaling is disturbed in the developing brain.

  17. Current measurements from SVP and APEX drifters around the Hawaiian Islands deployed by the NOAA Coral Reef Ecosystems Division (CRED), 2001-2006 (NODC Accession 0049436)

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — This data collection includes data from multiple SVP and APEX drifters deployed in the region around the Hawaiian Islands to assess ocean currents and sea surface...

  18. Cone Beam Computed Tomography Evaluation of the Periapical Status of Nonvital Tooth with Open Apex Obturated with Mineral Trioxide Aggregate: A Case Report

    Directory of Open Access Journals (Sweden)

    Vijay Shekhar

    2013-01-01

    Full Text Available Management of a tooth with open apex is a challenge to the dental practitioners. Evaluation of the periapical healing is required in such cases by radiographic techniques. The objective of this paper was to assess the healing of a periapical lesion in a non-vital tooth with open apex treated with mineral trioxide aggregate (MTA obturation using cone beam computed tomography (CBCT. The endodontic treatment of a fractured non-vital discolored maxillary left lateral incisor with an open apex was done with MTA obturation. The clinical and radiographic followup done regularly showed that the tooth was clinically asymptomatic and that the size of the periapical lesion observed by intraoral periapical (IOPA radiographs and CBCT was decreased remarkably after two years. CBCT and IOPA radiographs were found to be useful radiographic tools to assess the healing of a large periapical lesion in a non-vital tooth with open apex managed by MTA obturation.

  19. Male CD81 Knockout Genotype Disrupts Mendelian Distribution of Offspring

    OpenAIRE

    Mordica, Whitney J.; Gallagher, Ryan J; Kennedy, Jenna L; Chapes, Stephen K.

    2010-01-01

    CD81 is an integral membrane protein in the tetraspanin superfamily that serves as an adaptor protein. CD81 is also a maternally imprinted gene that is found in a regulated cluster of genes on mouse chromosome 7. Among offspring produced from heterozygous breeding pairs, CD81null/null mice grew at the same rate as CD81+/+ and CD81+/null mice. Because of an inhibition in sperm–egg fusion, CD81null/null female mice are much less fertile than CD81+/+ and CD81+/null mice. However, no published st...

  20. A comparative investigation on the accuracy of Neosono Ultima EZ instrument, radiography and anexperimental apex locator in root canal working length measurement

    OpenAIRE

    Zarabian M.; Nekoofar M.H; Marashi MS

    2004-01-01

    Various methods have already been suggested to measure the working length in root"ncanal therapy among which the use of paper point, tactile sense, radiography and electronic apex finder"ninstruments can be named. Investigations on EAL (Electronic Apex Locator) indicate that these instruments"ncan reduce the number of required radiography to a great extent. Moreover, in special cases such as patients"nwith high risk reflex, pregnant women, the mental disabled and handicapp...

  1. C-Terminal-Truncated Microdystrophin Recruits Dystrobrevin and Syntrophin to the Dystrophin-Associated Glycoprotein Complex and Reduces Muscular Dystrophy in Symptomatic Utrophin/Dystrophin Double-Knockout Mice

    OpenAIRE

    Yue, Yongping; LIU, MINGJU; Duan, Dongsheng

    2006-01-01

    C-terminal-truncated (ΔC) microdystrophin is being developed for Duchenne muscular dystrophy gene therapy. Encouraging results have been achieved in the mdx mouse model. Unfortunately, mdx mice do not display the same phenotype as human patients. Evaluating ΔC microdystrophin in a symptomatic model will be of significant relevance to human trials. Utrophin/dystrophin double-knockout (u-dko) mice were developed to model severe dystrophic changes in human patients. In this study we evaluated th...

  2. Comparative proteomics analysis of apoptotic Spodoptera frugiperda cells during p35 knockout Autographa californica multiple nucleopolyhedrovirus infection.

    Science.gov (United States)

    Yu, Qian; Xiong, Youhua; Liu, Jianliang; Wang, Qin; Qiu, Yuanxin; Wen, Dongling

    2016-06-01

    Infection with Autographa californica multiple nucleopolyhedrovirus (AcMNPV) mutants lacking a functional p35 gene can induce host cell apoptosis, which provides the possibility to use the potential of these viruses in the biological control of pest insects. Nonetheless, the proteomics or the protein changes of Spodoptera frugiperda (Sf9) cells infected with p35 knockout AcMNPV have not yet been studied. To further improve the use of AcMNPV, we set out to analyze the protein composition and protein changes of Sf9 cells of different infection stages by isobaric tag for relative and absolute quantification (iTRAQ) techniques. A total of 4004 sf9 proteins were identified by iTRAQ. After comparation of the significantly expressed 483 proteins from p35koAcMNPV-infected Sf9 cells and the significantly expressed 413 proteins from wtAcMNPV-infected Sf9 cells, we found that 226 proteins were specific to p35koAcMNPV-infected Sf9 cells. The 226 proteins were categorized according to GO classification for insects and were categorized into: biological processes, molecular functions and cellular components. Of interest, the most up-regulated proteins related to Epstein-Barr virus infection, RNA transport, Calcium signaling pathway, cGMP-PKG signaling pathway, oxidative phosphorylation and N-Glycan biosynthesis. Determination of the protein changes in p35 knockout AcMNPV-infected Sf9 cells would facilitate the better use of this virus-host cell interaction in pest insect control and other related fields. PMID:26922645

  3. Serotonin abnormalities in Engrailed-2 knockout mice: New insight relevant for a model of Autism Spectrum Disorder.

    Science.gov (United States)

    Viaggi, Cristina; Gerace, Claudio; Pardini, Carla; Corsini, Giovanni U; Vaglini, Francesca

    2015-08-01

    Autism spectrum disorder (ASD) is a congenital neurodevelopmental behavioral disorder that appears in early childhood. Recent human genetic studies identified the homeobox transcription factor, Engrailed 2 (EN2), as a possible ASD susceptibility gene. En2 knockout mice (En2-/-) display subtle cerebellar neuropathological changes and reduced levels of tyrosine hydroxylase, noradrenaline and serotonin in the hippocampus and cerebral cortex similar to those ones which have been observed in the ASD brain. Furthermore other similarities link En2 knockout mice to ASD patients. Several lines of evidence suggest that serotonin may play an important role in the pathophysiology of the disease. In the present study we measured, by using an HPLC, the 5-HT levels in different brain areas and at different ages in En2-/- mice. In the frontal and occipital cortex, the content of 5HT was reduced in En2-/- 1 and 3 months old mice; in 6 month old mice, the difference was still present, but it was not statistically significant. The 5-HT content of cerebellar cortex was significantly reduced at 1 month old but significantly high when the KO mice reached 3 months of age. The increase was present even at 6 months of age. A similar trend was highlighted by SERT immunolabeling in En2-/- mice compared to control in the same areas and age analyzed. Our findings, in agreement with the current knowledge on the 5-HT system alterations in ASD, confirm the early neurotransmitter deficit with a late compensatory recovery in En2 KO-mice further suggesting that this experimental animal may be considered a good predictive model for the human disease. PMID:26002543

  4. Generation and characterization of an Nxf7 knockout mouse to study NXF5 deficiency in a patient with intellectual disability.

    Directory of Open Access Journals (Sweden)

    Lieselot Vanmarsenille

    Full Text Available Members of the Nuclear eXport Factor (NXF family are involved in the export of mRNA from the nucleus to the cytoplasm, or hypothesized to play a role in transport of cytoplasmic mRNA. We previously reported on the loss of NXF5 in a male patient with a syndromic form of intellectual disability. To study the functional role of NXF5 we identified the mouse counterpart. Based on synteny, mouse Nxf2 is the ortholog of human NXF5. However, we provide several lines of evidence that mouse Nxf7 is the actual functional equivalent of NXF5. Both Nxf7 and NXF5 are predominantly expressed in the brain, show cytoplasmic localization, and present as granules in neuronal dendrites suggesting a role in cytoplasmic mRNA metabolism in neurons. Nxf7 was primarily detected in the pyramidal cells of the hippocampus and in layer V of the cortex. Similar to human NXF2, mouse Nxf2 is highly expressed in testis and shows a nuclear localization. Interestingly, these findings point to a different evolutionary path for both NXF genes in human and mouse. We thus generated and validated Nxf7 knockout mice, which were fertile and did not present any gross anatomical or morphological abnormalities. Expression profiling in the hippocampus and the cortex did not reveal significant changes between wild-type and Nxf7 knockout mice. However, impaired spatial memory was observed in these KO mice when evaluated in the Morris water maze test. In conclusion, our findings provide strong evidence that mouse Nxf7 is the functional counterpart of human NXF5, which might play a critical role in mRNA metabolism in the brain.

  5. Fronto-temporo-orbitozygomatic craniotomy and "half-and-half" approach for basilar apex aneurysms

    Directory of Open Access Journals (Sweden)

    Behari Sanjay

    2009-01-01

    Full Text Available Background: Basilar apex aneurysms (BAA are located in interpeduncular cistern surrounded by eloquent neurovascular structures. Surgical access is difficult due to narrow surgical corridors and requires traversing through a depth of 6-8 cm of subarachnoid space. Aim: Surgical management of BAAs clipped using frontotemporal craniotomy, orbitozygomatic osteotomy with combined subtemporal and transylvian (half and half approach is discussed. Setting and Design: Tertiary care referral institute; prospective study. Materials and Methods: Five patients with BAA rupture causing subarachnoid hemorrhage presented in modified Hunt and Hess (Hand H grades II (n=1, III (n=1 and IV (n=3, respectively. In 4 patients, the aneurysms were 0.8-1.2 cm in diameter, situated 7 mm-1 cm above dorsum sellae. Two of them had posteriorly projecting aneurysms. One patient had a giant, high BAA with a left parietooccipital arteriovenous malformation. Vasospasm of posterior cerebral/proximal basilar artery was seen in 2 patients. In one patient, internal carotid artery was mobilized by intradural anterior clinoid drilling with carotid collar division. Triple-H therapy was administered following surgery. Results: There was no intraoperative rupture or temporary clipping. Follow up angiography showed complete aneurysmal obliteration with preservation of posterior cerebral and superior cerebellar arteries. Follow up (mean: 8.7 ± 3.5 months H and H grades were II (n=2 and III (n=3, respectively. The morbidity include caudate and thalamic region infarct, transient III rd nerve palsy and cerebrospinal fluid otorrhoea (n=1, respectively. Conclusions: This simple approach provides a wide surgical corridor from 5 mm below to greater than 1 cm above dorsum sellae with adequate proximal control of basilar artery. It is an option to endovascular embolization especially with large and giant, or wide-necked BAA, vertebrobasilar tortuosity, coil compaction or postcoiling re-rupture and

  6. Climate change alters the trophic niche of a declining apex marine predator.

    Science.gov (United States)

    Bond, Alexander L; Lavers, Jennifer L

    2014-07-01

    Changes in the world's oceans have altered nutrient flow, and affected the viability of predator populations when prey species become unavailable. These changes are integrated into the tissues of apex predators over space and time and can be quantified using stable isotopes in the inert feathers of historical and contemporary avian specimens. We measured δ(13) C and δ(15) N values in Flesh-footed Shearwaters (Puffinus carneipes) from Western and South Australia from 1936-2011. The Flesh-footed Shearwaters more than doubled their trophic niche (from 3.91 ± 1.37 ‰(2) to 10.00 ± 1.79 ‰(2) ), and dropped an entire trophic level in 75 years (predicted δ(15) N decreased from +16.9 ‰ to + 13.5 ‰, and δ(13) C from -16.9 ‰ to -17.9 ‰) - the largest change in δ(15) N yet reported in any marine bird, suggesting a relatively rapid shift in the composition of the Indian Ocean food web, or changes in baseline δ(13) C and δ(15) N values. A stronger El Niño-Southern Oscillation results in a weaker Leeuwin Current in Western Australia, and decreased Flesh-footed Shearwater δ(13) C and δ(15) N. Current climate forecasts predict this trend to continue, leading to increased oceanic 'tropicalization' and potentially competition between Flesh-footed Shearwaters and more tropical sympatric species with expanding ranges. Flesh-footed Shearwater populations are declining, and current conservation measures aimed primarily at bycatch mitigation are not restoring populations. Widespread shifts in foraging, as shown here, may explain some of the reported decline. An improved understanding and ability to mitigate the impacts of global climactic changes is therefore critical to the long-term sustainability of this declining species. PMID:24615959

  7. A HERSCHEL AND APEX CENSUS OF THE REDDEST SOURCES IN ORION: SEARCHING FOR THE YOUNGEST PROTOSTARS

    International Nuclear Information System (INIS)

    We perform a census of the reddest, and potentially youngest, protostars in the Orion molecular clouds using data obtained with the PACS instrument on board the Herschel Space Observatory and the LABOCA and SABOCA instruments on APEX as part of the Herschel Orion Protostar Survey (HOPS). A total of 55 new protostar candidates are detected at 70 μm and 160 μm that are either too faint (m24 > 7 mag) to be reliably classified as protostars or undetected in the Spitzer/MIPS 24 μm band. We find that the 11 reddest protostar candidates with log λFλ70/λFλ24 > 1.65 are free of contamination and can thus be reliably explained as protostars. The remaining 44 sources have less extreme 70/24 colors, fainter 70 μm fluxes, and higher levels of contamination. Taking the previously known sample of Spitzer protostars and the new sample together, we find 18 sources that have log λFλ70/λFλ24 > 1.65; we name these sources 'PACS Bright Red sources', or PBRs. Our analysis reveals that the PBR sample is composed of Class 0 like sources characterized by very red spectral energy distributions (SEDs; Tbol smm/Lbol > 0.6%). Modified blackbody fits to the SEDs provide lower limits to the envelope masses of 0.2-2 M☉ and luminosities of 0.7-10 L☉. Based on these properties, and a comparison of the SEDs with radiative transfer models of protostars, we conclude that the PBRs are most likely extreme Class 0 objects distinguished by higher than typical envelope densities and hence, high mass infall rates.

  8. A HERSCHEL AND APEX CENSUS OF THE REDDEST SOURCES IN ORION: SEARCHING FOR THE YOUNGEST PROTOSTARS

    Energy Technology Data Exchange (ETDEWEB)

    Stutz, Amelia M.; Robitaille, Thomas; Henning, Thomas; Krause, Oliver [Max Planck Institute for Astronomy, Koenigstuhl 17, D-69117 Heidelberg (Germany); Tobin, John J. [National Radio Astronomy Observatory, Charlottesville, VA 22903 (United States); Stanke, Thomas [ESO, Karl-Schwarzschild-Strasse 2, D-85748 Garching bei Muenchen (Germany); Megeath, S. Thomas; Fischer, William J. [Department of Physics and Astronomy, University of Toledo, 2801 W. Bancroft Street, Toledo, OH 43606 (United States); Ali, Babar; Furlan, Elise [NHSC/IPAC/Caltech, 770 S. Wilson Avenue, Pasadena, CA 91125 (United States); Di Francesco, James [National Research Council of Canada, Herzberg Institute of Astrophysics, 5071 West Saanich Road, Victoria, BC V9E 2E7 (Canada); Hartmann, Lee [Department of Astronomy, University of Michigan, 830 Dennison Building, 500 Church Street, Ann Arbor, MI 48109 (United States); Osorio, Mayra [Instituto de Astrofisica de Andalucia, CSIC, Camino Bajo de Huetor 50, E-18008 Granada (Spain); Wilson, Thomas L. [Naval Research Laboratory, 4555 Overlook Avenue SW, Washington, DC 20375 (United States); Allen, Lori [National Optical Astronomy Observatory, 950 North Cherry Avenue, Tucson, AZ 85719 (United States); Manoj, P., E-mail: stutz@mpia.de [Department of Physics and Astronomy, 500 Wilson Boulevard, University of Rochester, Rochester, NY 14627 (United States)

    2013-04-10

    We perform a census of the reddest, and potentially youngest, protostars in the Orion molecular clouds using data obtained with the PACS instrument on board the Herschel Space Observatory and the LABOCA and SABOCA instruments on APEX as part of the Herschel Orion Protostar Survey (HOPS). A total of 55 new protostar candidates are detected at 70 {mu}m and 160 {mu}m that are either too faint (m{sub 24} > 7 mag) to be reliably classified as protostars or undetected in the Spitzer/MIPS 24 {mu}m band. We find that the 11 reddest protostar candidates with log {lambda}F{sub {lambda}}70/{lambda}F{sub {lambda}}24 > 1.65 are free of contamination and can thus be reliably explained as protostars. The remaining 44 sources have less extreme 70/24 colors, fainter 70 {mu}m fluxes, and higher levels of contamination. Taking the previously known sample of Spitzer protostars and the new sample together, we find 18 sources that have log {lambda}F{sub {lambda}}70/{lambda}F{sub {lambda}}24 > 1.65; we name these sources 'PACS Bright Red sources', or PBRs. Our analysis reveals that the PBR sample is composed of Class 0 like sources characterized by very red spectral energy distributions (SEDs; T{sub bol} < 45 K) and large values of sub-millimeter fluxes (L{sub smm}/L{sub bol} > 0.6%). Modified blackbody fits to the SEDs provide lower limits to the envelope masses of 0.2-2 M{sub Sun} and luminosities of 0.7-10 L{sub Sun }. Based on these properties, and a comparison of the SEDs with radiative transfer models of protostars, we conclude that the PBRs are most likely extreme Class 0 objects distinguished by higher than typical envelope densities and hence, high mass infall rates.

  9. Investigation of radiation damage in structural material of APEX reactor by using Monte Carlo method

    International Nuclear Information System (INIS)

    Highlights: ► In this study was calculated effects of the radiation damage on the selected fluid rates and thicknesses of liquid second wall. ► MCNPX-2.5.0 Monte Carlo code was used for three-dimensional calculations. ► The heavy metal was found as appropriate 10% ThF4 for DPA and 10% UO2 for gas production. ► DPA in the structural material was calculated as appropriate for 40–50 cm thickness of the liquid second wall. - Abstract: In this study, ThF4, UF4 and UO2 heavy metals were used with ratios of 2% and 10% while Flibe (Li2BeF4) molten salt mixture and 100% Flibe were used as fluids in the liquid first wall, liquid second wall and shield zones of the APEX. The steel wall that is used as a structural material is 4 cm in thickness and calculations for each 0.5 cm thick zone of the steel wall were performed. In this study, the total changes caused by radiation damage in the steel used as a structural material for 20 cm, 30 cm, 40 cm and 50 cm thicknesses of liquid second wall, for each 0.5 cm thick zones of the steel wall and for the selected fluid rates were investigated for 30 full power years (FPY). The neutron wall load is assumed to be 10 MW/m2. Three-dimensional nucleonic calculations were performed using MCNPX-2.5.0 Monte Carlo code and ENDF/B-VI nuclear data library

  10. Effects of D1 receptor knockout on fear and reward learning.

    Science.gov (United States)

    Abraham, Antony D; Neve, Kim A; Lattal, K Matthew

    2016-09-01

    Dopamine signaling is involved in a variety of neurobiological processes that contribute to learning and memory. D1-like dopamine receptors (including D1 and D5 receptors) are thought to be involved in memory and reward processes, but pharmacological approaches have been limited in their ability to distinguish between D1 and D5 receptors. Here, we examine the effects of a specific knockout of D1 receptors in associative learning tasks involving aversive (shock) or appetitive (cocaine) unconditioned stimuli. We find that D1 knockout mice show similar levels of cued and contextual fear conditioning to WT controls following conditioning protocols involving one, two, or four shocks. D1 knockout mice show increased generalization of fear conditioning and extinction across contexts, revealed as increased freezing to a novel context following conditioning and decreased freezing to an extinguished cue during a contextual renewal test. Further, D1 knockout mice show mild enhancements in extinction following an injection of SKF81297, a D1/D5 receptor agonist, suggesting a role for D5 receptors in extinction enhancements induced by nonspecific pharmacological agonists. Finally, although D1 knockout mice show decreased locomotion induced by cocaine, they are able to form a cocaine-induced conditioned place preference. We discuss these findings in terms of the role of dopamine D1 receptors in general learning and memory processes. PMID:27423521

  11. Atherosclerosis, inflammation and lipoprotein glomerulopathy in kidneys of apoE-/-/LDL-/- double knockout mice

    Directory of Open Access Journals (Sweden)

    Langheinrich Alexander C

    2010-08-01

    Full Text Available Abstract Background The apoE-/-/LDL-/- double knockout mice are bearing considerable structural homology to human atherosclerosis. We hypothesized, that advanced lesion formation in the renal artery is associated with kidney alterations in these mice. Methods Kidneys from apoE-/-/LDL-/- double knockout mice at the age of 80 weeks (n = 6 and C57/BL control mice (n = 5 were infused with Microfil, harvested and scanned with micro-CT (12 μm cubic voxels and Nano-CT (900 nm cubic voxels. We quantitated the total vascular volume using micro-CT. Number and cross-sectional area (μm2 of glomeruli were measured using histology. Results At the age of 80 weeks, the renal total vascular volume fraction decreased significantly (p -/-/LDL-/- double knockout mice, predominantly involved are plasma cells and leucocytes. Glomeruli cross-sectional area (9959 ± 1083 μm2 and number (24.8 ± 4.5 increased in apoE-/-/LDL-/- double knockout mice compared to controls (3533 ± 398 μm2; 17.6 ± 3, respectively, whereas 41% of the total number of glomeruli showed evidence for lipoprotein associated glomerulopathy (LPG. Moreover, immunohistochemistry demonstrated capillary aneurysms of the glomeruli filled with factor 8 containing emboli. Conclusion The reduced intra-renal total vascular volume is associated with systemic atherosclerosis and glomeruli alterations in the apoE-/-/LDL-/- double knockout mouse model.

  12. Aggravated renal inflammatory responses in TRPV1 gene knockout mice subjected to DOCA-salt hypertension

    OpenAIRE

    Wang, Youping; Wang, Donna H.

    2009-01-01

    To test the hypothesis that deletion of the transient receptor potential vanilloid type 1 (TRPV1) channel exaggerates hypertension-induced renal inflammatory response, wild-type (WT) or TRPV1-null mutant (TRPV1−/−) mice were subjected to uninephrectomy and deoxycorticosterone acetate (DOCA)-salt treatment for 4 wk. Mean arterial pressure (MAP) determined by radiotelemetry increased in DOCA-salt-treated WT or TRPV1−/− mice, whereas there was no difference in MAP between two strains at the base...

  13. Restoration of Hearing in the VGLUT3 Knockout Mouse Using Virally-Mediated Gene Therapy

    OpenAIRE

    Akil, Omar; Seal, Rebecca P.; Burke, Kevin; Wang, Chuansong; Alemi, Aurash; During, Matthew; Edwards, Robert H.; Lustig, Lawrence R.

    2012-01-01

    Mice lacking the vesicular glutamate transporter-3 (VGLUT3) are congenitally deaf due to loss of glutamate release at the inner hair cell afferent synapse. Cochlear delivery of VGLUT3 using adeno-associated virus-1 (AAV1) leads to transgene expression in only inner hair cells (IHC), despite broader viral uptake. Within two weeks of AAV1-VGLUT3 delivery, acoustic brainstem response (ABR) thresholds normalize, along with partial rescue of the startle response. Lastly, we demonstrate partial rev...

  14. 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; Andreelli, Fabrizio; Birk, Jesper Bratz; Hellsten, Ylva; Schjerling, Peter; Vaulont, Sophie; Neufer, P. Darrell; Richter, Erik A.; Pilegaard, Henriette

    2005-01-01

    did not affect exercise-induced transcription. Exercise increased the mRNA content of PGC-1a, Forkhead box class O (FOXO)1, HKII, and pyruvate dehydrogenase kinase 4 (PDK4) similarly in a2-WT and a2-KO mice, whereas glucose transporter GLUT 4, carnitine palmitoyltransferase 1 (CPTI), lipoprotein...

  15. Determination of tissue and cell function of Nuclear Myosin 1 by gene knock-out

    Czech Academy of Sciences Publication Activity Database

    Venit, Tomáš; Dzijak, Rastislav; Kahle, Michal; Hozák, Pavel

    Gothenburg : FEBS, 2010. s. 24-24. [FEBS Congress /35./ - Molecules of Life. 26.06.10-01.07.10, Gothenburg] Institutional research plan: CEZ:AV0Z50520514 Subject RIV: EB - Genetics ; Molecular Biology

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

    Science.gov (United States)

    The North American opossum (Didelphis virginiana) is the definitive host for at least three named species of Sarcocystis: S. falcatula, S. neurona, and S. speeri. It appears that there may be additional undescribed species of Sarcocystis in D. virginiana feces. The South American opossums (D. albive...

  17. Dose-dependent effects of UVB-induced skin carcinogenesis in hairless p53 knockout mice

    International Nuclear Information System (INIS)

    Exposure to (solar) UVB radiation gives rise to mutations in the p53 tumor suppressor gene that appear to contribute to the earliest steps in the molecular cascade towards human and murine skin cancer. To examine in more detail the role of p53, we studied UVB-induced carcinogenesis in hairless p53 knock-out mice. The early onset of lymphomas as well as early wasting of mice interfered with the development of skin tumors in p53 null-mice. The induction of skin tumors in the hairless p53+/- mice was accomplished by daily exposure to two different UV-doses of approximately 450 J/m2 and 900 J/m2 from F40 lamps corresponding to a fraction of about 0.4 and 0.8 of the minimal edemal dose. Marked differences in skin carcinogenesis were observed between the p53+/- mice and their wild type littermates. Firstly, at 900 J/m2, tumors developed significantly faster in the heterozygotes than in wild types, whereas at 450 J/m2 there was hardly any difference, suggesting that only at higher damage levels loss of one functional p53 allele is important. Secondly, a large portion (25%) of skin tumors in the heterozygotes were of a more malignant, poorly differentiated variety of squamous cell carcinomas, i.e. spindle cell carcinomas, a tumor type that was rarely observed in daily UV exposed wild type hairless mice. Thirdly, the p53 mutation spectrum in skin tumors in heterozygotes is quite different from that in wild types. Together these results support the notion that a point mutation in the p53 gene impacts skin carcinogenesis quite differently than allelic loss: the former is generally selected for in early stages of skin tumors in wild type mice, whereas the latter enhances tumor development only at high exposure levels (where apoptosis becomes more prevalent) and appears to increase progression (to a higher grade of malignancy) of skin tumors

  18. The Epstein-Barr Virus BDLF4 Gene Is Required for Efficient Expression of Viral Late Lytic Genes.

    Science.gov (United States)

    Watanabe, Takahiro; Narita, Yohei; Yoshida, Masahiro; Sato, Yoshitaka; Goshima, Fumi; Kimura, Hiroshi; Murata, Takayuki

    2015-10-01

    Epstein-Barr virus (EBV) is a gammaherpesvirus, associated with infectious mononucleosis and various types of malignancy. We focused here on the BDLF4 gene of EBV and identified it as a lytic gene, expressed with early kinetics. Viral late gene expression of the BDLF4 knockout strain was severely restricted; this could be restored by an exogenous supply of BDLF4. These results indicate that BDLF4 is important for the EBV lytic replication cycle, especially in late gene expression. PMID:26202235

  19. High-temperature expansion and knock-out properties of moulding sands with water glass

    Directory of Open Access Journals (Sweden)

    Major-Gabryś K.

    2007-01-01

    Full Text Available The article focuses on the topic of improving the knock-out properties of moulding sand with water glass and ester hardener. It is settled that the cause of worse knock-out properties of moulding sand can be brought by their thermal expansion in increased temperatures. There is a presentation of the influence of different additives, containing Al2O3, on moulding sands’ expansion in increased temperatures. Within the frames of research, there was an elaboration of the influence of authors own additive- Glassex, on the expansion phenomenon of moulding sands with water glass and ester hardener. It is concluded, that the new additive stops the expansion of moulding sands and as well it improves their knock-out properties.

  20. MR histology of advanced atherosclerotic lesions of ApoE- knockout mice

    Science.gov (United States)

    Naumova, A.; Yarnykh, V.; Ferguson, M.; Rosenfeld, M.; Yuan, C.

    2016-02-01

    The purposes of this study were to examine the feasibility of determining the composition of advanced atherosclerotic plaques in fixed ApoE-knockout mice and to develop a time-efficient microimaging protocol for MR histological imaging on mice. Five formalin-fixed transgenic ApoE-knockout mice were imaged at the 9.4T Bruker BioSpec MR scanner using 3D spoiled gradient-echo sequence with an isotropic field of view of 24 mm3; TR 20.8 ms; TE 2.6 ms; flip angle 20°, resulted voxel size 47 × 63 × 94 pm3. MRI examination has shown that advanced atherosclerotic lesions of aorta, innominate and carotid arteries in ApoE-knockout mice are characterized by high calcification and presence of the large fibrofatty nodules. MRI quantification of atherosclerotic lesion components corresponded to histological assessment of plaque composition with a correlation coefficient of 0.98.