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Sample records for giy-yig homing endonuclease

  1. Divalent metal ion differentially regulates the sequential nicking reactions of the GIY-YIG homing endonuclease I-BmoI.

    Directory of Open Access Journals (Sweden)

    Benjamin P Kleinstiver

    Full Text Available Homing endonucleases are site-specific DNA endonucleases that function as mobile genetic elements by introducing double-strand breaks or nicks at defined locations. Of the major families of homing endonucleases, the modular GIY-YIG endonucleases are least understood in terms of mechanism. The GIY-YIG homing endonuclease I-BmoI generates a double-strand break by sequential nicking reactions during which the single active site of the GIY-YIG nuclease domain must undergo a substantial reorganization. Here, we show that divalent metal ion plays a significant role in regulating the two independent nicking reactions by I-BmoI. Rate constant determination for each nicking reaction revealed that limiting divalent metal ion has a greater impact on the second strand than the first strand nicking reaction. We also show that substrate mutations within the I-BmoI cleavage site can modulate the first strand nicking reaction over a 314-fold range. Additionally, in-gel DNA footprinting with mutant substrates and modeling of an I-BmoI-substrate complex suggest that amino acid contacts to a critical GC-2 base pair are required to induce a bottom-strand distortion that likely directs conformational changes for reaction progress. Collectively, our data implies mechanistic roles for divalent metal ion and substrate bases, suggesting that divalent metal ion facilitates the re-positioning of the GIY-YIG nuclease domain between sequential nicking reactions.

  2. Phylogenomic analysis of the GIY-YIG nuclease superfamily

    Directory of Open Access Journals (Sweden)

    Bujnicki Janusz M

    2006-04-01

    Full Text Available Abstract Background The GIY-YIG domain was initially identified in homing endonucleases and later in other selfish mobile genetic elements (including restriction enzymes and non-LTR retrotransposons and in enzymes involved in DNA repair and recombination. However, to date no systematic search for novel members of the GIY-YIG superfamily or comparative analysis of these enzymes has been reported. Results We carried out database searches to identify all members of known GIY-YIG nuclease families. Multiple sequence alignments together with predicted secondary structures of identified families were represented as Hidden Markov Models (HMM and compared by the HHsearch method to the uncharacterized protein families gathered in the COG, KOG, and PFAM databases. This analysis allowed for extending the GIY-YIG superfamily to include members of COG3680 and a number of proteins not classified in COGs and to predict that these proteins may function as nucleases, potentially involved in DNA recombination and/or repair. Finally, all old and new members of the GIY-YIG superfamily were compared and analyzed to infer the phylogenetic tree. Conclusion An evolutionary classification of the GIY-YIG superfamily is presented for the very first time, along with the structural annotation of all (subfamilies. It provides a comprehensive picture of sequence-structure-function relationships in this superfamily of nucleases, which will help to design experiments to study the mechanism of action of known members (especially the uncharacterized ones and will facilitate the prediction of function for the newly discovered ones.

  3. Phage T4 endonuclease SegD that is similar to group I intron endonucleases does not initiate homing of its own gene.

    Science.gov (United States)

    Sokolov, Andrey S; Latypov, Oleg R; Kolosov, Peter M; Shlyapnikov, Michael G; Bezlepkina, Tamara A; Kholod, Natalia S; Kadyrov, Farid A; Granovsky, Igor E

    2018-02-01

    Homing endonucleases are a group of site-specific endonucleases that initiate homing, a nonreciprocal transfer of its own gene into a new allele lacking this gene. This work describes a novel phage T4 endonuclease, SegD, which is homologous to the GIY-YIG family of homing endonucleases. Like other T4 homing endonucleases SegD recognizes an extended, 16bp long, site, cleaves it asymmetrically to form 3'-protruding ends and digests both unmodified DNA and modified T-even phage DNA with similar efficiencies. Surprisingly, we revealed that SegD cleavage site was identical in the genomes of segD - and segD + phages. We found that segD gene was expressed during the T4 developmental cycle. Nevertheless, endonuclease SegD was not able to initiate homing of its own gene as well as genetic recombination between phages in its site inserted into the rII locus. Copyright © 2018 Elsevier Inc. All rights reserved.

  4. Phage T4 SegB protein is a homing endonuclease required for the preferred inheritance of T4 tRNA gene region occurring in co-infection with a related phage.

    Science.gov (United States)

    Brok-Volchanskaya, Vera S; Kadyrov, Farid A; Sivogrivov, Dmitry E; Kolosov, Peter M; Sokolov, Andrey S; Shlyapnikov, Michael G; Kryukov, Valentine M; Granovsky, Igor E

    2008-04-01

    Homing endonucleases initiate nonreciprocal transfer of DNA segments containing their own genes and the flanking sequences by cleaving the recipient DNA. Bacteriophage T4 segB gene, which is located in a cluster of tRNA genes, encodes a protein of unknown function, homologous to homing endonucleases of the GIY-YIG family. We demonstrate that SegB protein is a site-specific endonuclease, which produces mostly 3' 2-nt protruding ends at its DNA cleavage site. Analysis of SegB cleavage sites suggests that SegB recognizes a 27-bp sequence. It contains 11-bp conserved sequence, which corresponds to a conserved motif of tRNA TpsiC stem-loop, whereas the remainder of the recognition site is rather degenerate. T4-related phages T2L, RB1 and RB3 contain tRNA gene regions that are homologous to that of phage T4 but lack segB gene and several tRNA genes. In co-infections of phages T4 and T2L, segB gene is inherited with nearly 100% of efficiency. The preferred inheritance depends absolutely on the segB gene integrity and is accompanied by the loss of the T2L tRNA gene region markers. We suggest that SegB is a homing endonuclease that functions to ensure spreading of its own gene and the surrounding tRNA genes among T4-related phages.

  5. Engineering a Nickase on the Homing Endonuclease I-DmoI Scaffold

    DEFF Research Database (Denmark)

    Molina, Rafael; Marcaida, María José; Redondo, Pilar

    2015-01-01

    strand break could be an approach to reduce the toxicity associated with non-homologous end joining by promoting the use of homologous recombination to repair the cleavage of a single DNA break. Taking advantage of the sequential DNA cleavage mechanism of I-DmoI LAGLIDADG homing endonuclease, we have......Homing endonucleases are useful tools for genome modification because of their capability to recognize and cleave specifically large DNA targets. These endonucleases generate a DNA double strand break that can be repaired by the DNA damage response machinery. The break can be repaired by homologous...

  6. Evolutionary maintenance of selfish homing endonuclease genes in the absence of horizontal transfer.

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    Yahara, Koji; Fukuyo, Masaki; Sasaki, Akira; Kobayashi, Ichizo

    2009-11-03

    Homing endonuclease genes are "selfish" mobile genetic elements whose endonuclease promotes the spread of its own gene by creating a break at a specific target site and using the host machinery to repair the break by copying and inserting the gene at this site. Horizontal transfer across the boundary of a species or population within which mating takes place has been thought to be necessary for their evolutionary persistence. This is based on the assumption that they will become fixed in a host population, where opportunities of homing will disappear, and become susceptible to degeneration. To test this hypothesis, we modeled behavior of a homing endonuclease gene that moves during meiosis through double-strand break repair. We mathematically explored conditions for persistence of the homing endonuclease gene and elucidated their parameter dependence as phase diagrams. We found that, if the cost of the pseudogene is lower than that of the homing endonuclease gene, the 2 forms can persist in a population through autonomous periodic oscillation. If the cost of the pseudogene is higher, 2 types of dynamics appear that enable evolutionary persistence: bistability dependent on initial frequency or fixation irrespective of initial frequency. The prediction of long persistence in the absence of horizontal transfer was confirmed by stochastic simulations in finite populations. The average time to extinction of the endonuclease gene was found to be thousands of meiotic generations or more based on realistic parameter values. These results provide a solid theoretical basis for an understanding of these and other extremely selfish elements.

  7. Crystal Structure of the Homing Endonuclease I-CvuI Provides a New Template for Genome Modification

    DEFF Research Database (Denmark)

    Molina, Rafael; Redondo, Pilar; López-Méndez, Blanca

    2015-01-01

    Homing endonucleases recognize and generate a DNA double-strand break, which has been used to promote gene targeting. These enzymes recognize long DNA stretches; they are highly sequence-specific enzymes and display a very low frequency of cleavage even in complete genomes. Although a large number...... of homing endonucleases have been identified, the landscape of possible target sequences is still very limited to cover the complexity of the whole eukaryotic genome. Therefore, the finding and molecular analysis of homing endonucleases identified but not yet characterized may widen the landscape...

  8. Home and away- the evolutionary dynamics of homing endonucleases

    Directory of Open Access Journals (Sweden)

    Barzel Adi

    2011-11-01

    Full Text Available Abstract Background Homing endonucleases (HEases are a large and diverse group of site-specific DNAases. They reside within self-splicing introns and inteins, and promote their horizontal dissemination. In recent years, HEases have been the focus of extensive research due to their promising potential use in gene targeting procedures for the treatment of genetic diseases and for the genetic engineering of crop, animal models and cell lines. Results Using mathematical analysis and computational modeling, we present here a novel account for the evolution and population dynamics of HEase genes (HEGs. We describe HEGs as paradoxical selfish elements whose long-term persistence in a single population relies on low transmission rates and a positive correlation between transmission efficiency and toxicity. Conclusion Plausible conditions allow HEGs to sustain at high frequency through long evolutionary periods, with the endonuclease frequency being either at equilibrium or periodically oscillating. The predictions of our model may prove important not only for evolutionary theory but also for gene therapy and bio-engineering applications of HEases.

  9. Evolution of divergent DNA recognition specificities in VDE homing endonucleases from two yeast species.

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    Posey, Karen L; Koufopanou, Vassiliki; Burt, Austin; Gimble, Frederick S

    2004-01-01

    Homing endonuclease genes (HEGs) are mobile DNA elements that are thought to confer no benefit to their host. They encode site-specific DNA endonucleases that perpetuate the element within a species population by homing and disseminate it between species by horizontal transfer. Several yeast species contain the VMA1 HEG that encodes the intein-associated VMA1-derived endonuclease (VDE). The evolutionary state of VDEs from 12 species was assessed by assaying their endonuclease activities. Only two enzymes are active, PI-ZbaI from Zygosaccharomyces bailii and PI-ScaI from Saccharomyces cariocanus. PI-ZbaI cleaves the Z.bailii recognition sequence significantly faster than the Saccharomyces cerevisiae site, which differs at six nucleotide positions. A mutational analysis indicates that PI-ZbaI cleaves the S.cerevisiae substrate poorly due to the absence of a contact that is analogous to one made in PI-SceI between Gln-55 and nucleotides +9/+10. PI-ZbaI cleaves the Z.bailii substrate primarily due to a single base-pair substitution (A/T+5 --> T/A+5). Structural modeling of the PI-ZbaI/DNA complex suggests that Arg-331, which is absent in PI-SceI, contacts T/A+5, and the reduced activity observed in a PI-ZbaI R331A mutant provides evidence for this interaction. These data illustrate that homing endonucleases evolve altered specificity as they adapt to recognize alternative target sites.

  10. Evolution of divergent DNA recognition specificities in VDE homing endonucleases from two yeast species

    Science.gov (United States)

    Posey, Karen L.; Koufopanou, Vassiliki; Burt, Austin; Gimble, Frederick S.

    2004-01-01

    Homing endonuclease genes (HEGs) are mobile DNA elements that are thought to confer no benefit to their host. They encode site-specific DNA endonucleases that perpetuate the element within a species population by homing and disseminate it between species by horizontal transfer. Several yeast species contain the VMA1 HEG that encodes the intein-associated VMA1-derived endonuclease (VDE). The evolutionary state of VDEs from 12 species was assessed by assaying their endonuclease activities. Only two enzymes are active, PI-ZbaI from Zygosaccharomyces bailii and PI-ScaI from Saccharomyces cariocanus. PI-ZbaI cleaves the Z.bailii recognition sequence significantly faster than the Saccharomyces cerevisiae site, which differs at six nucleotide positions. A mutational analysis indicates that PI-ZbaI cleaves the S.cerevisiae substrate poorly due to the absence of a contact that is analogous to one made in PI-SceI between Gln-55 and nucleotides +9/+10. PI-ZbaI cleaves the Z.bailii substrate primarily due to a single base-pair substitution (A/T+5 → T/A+5). Structural modeling of the PI-ZbaI/DNA complex suggests that Arg-331, which is absent in PI-SceI, contacts T/A+5, and the reduced activity observed in a PI-ZbaI R331A mutant provides evidence for this interaction. These data illustrate that homing endonucleases evolve altered specificity as they adapt to recognize alternative target sites. PMID:15280510

  11. Karyopherin-Mediated Nuclear Import of the Homing Endonuclease VMA1-Derived Endonuclease Is Required for Self-Propagation of the Coding Region

    OpenAIRE

    Nagai, Yuri; Nogami, Satoru; Kumagai-Sano, Fumi; Ohya, Yoshikazu

    2003-01-01

    VMA1-derived endonuclease (VDE), a site-specific endonuclease in Saccharomyces cerevisiae, enters the nucleus to generate a double-strand break in the VDE-negative allelic locus, mediating the self-propagating gene conversion called homing. Although VDE is excluded from the nucleus in mitotic cells, it relocalizes at premeiosis, becoming localized in both the nucleus and the cytoplasm in meiosis. The nuclear localization of VDE is induced by inactivation of TOR kinases, which constitute centr...

  12. Homing endonuclease genes: the rise and fall and rise again of a selfish element.

    Science.gov (United States)

    Burt, Austin; Koufopanou, Vassiliki

    2004-12-01

    Homing endonuclease genes (HEGs) are selfish genetic elements that spread by first cleaving chromosomes that do not contain them and then getting copied across to the broken chromosome as a byproduct of the repair process. The success of this strategy will depend on the opportunities for homing--in other words, the frequency with which HEG(+) and HEG(-) chromosomes come into contact--which varies widely among host taxa. HEGs are also unusual in that the selection pressure for endonuclease function disappears if they become fixed in a population, which makes them susceptible to degeneration and imposes a need for regular horizontal transmission between species. HEGs will be selected to reduce the harm done to the host organism, and this is expected to influence the evolution of their sequence specificity and maturase functions. HEGs may also be domesticated by their hosts, and are currently being put to human uses.

  13. Karyopherin-mediated nuclear import of the homing endonuclease VMA1-derived endonuclease is required for self-propagation of the coding region.

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    Nagai, Yuri; Nogami, Satoru; Kumagai-Sano, Fumi; Ohya, Yoshikazu

    2003-03-01

    VMA1-derived endonuclease (VDE), a site-specific endonuclease in Saccharomyces cerevisiae, enters the nucleus to generate a double-strand break in the VDE-negative allelic locus, mediating the self-propagating gene conversion called homing. Although VDE is excluded from the nucleus in mitotic cells, it relocalizes at premeiosis, becoming localized in both the nucleus and the cytoplasm in meiosis. The nuclear localization of VDE is induced by inactivation of TOR kinases, which constitute central regulators of cell differentiation in S. cerevisiae, and by nutrient depletion. A functional genomic approach revealed that at least two karyopherins, Srp1p and Kap142p, are required for the nuclear localization pattern. Genetic and physical interactions between Srp1p and VDE imply direct involvement of karyopherin-mediated nuclear transport in this process. Inactivation of TOR signaling or acquisition of an extra nuclear localization signal in the VDE coding region leads to artificial nuclear localization of VDE and thereby induces homing even during mitosis. These results serve as evidence that VDE utilizes the host systems of nutrient signal transduction and nucleocytoplasmic transport to ensure the propagation of its coding region.

  14. YIG magnonics

    International Nuclear Information System (INIS)

    Serga, A A; Chumak, A V; Hillebrands, B

    2010-01-01

    Early experiments in magnonics were made using ferrite samples, largely due to the intrinsically low magnetic (spin-wave) damping in these materials. Historically, magnonic phenomena were studied on micrometre to millimetre length scales. Today, the principal challenge in applied magnonics is to create sub-micrometre devices using modern polycrystalline magnetic alloys. However, until certain technical obstacles are overcome in these materials, ferrites-in particular yttrium iron garnet (YIG)-remain a valuable source of insight. At a time when interest in magnonic systems is particularly strong, it is both useful and timely to review the main scientific results of YIG magnonics of the last two decades, and to discuss the transferability of the concepts and ideas learned in ferrite materials to modern nano-scale systems.

  15. Surface modification of YIG by magnet array

    International Nuclear Information System (INIS)

    Atalay, S.; Kolat, V.S.; Bakır, H.G.; Izgi, T.; Kaya, A.O.; Kaya, O.A.; Gencer, H.

    2015-01-01

    Highlights: • The surface of YIG films were magnetically modulated by magnet array. • The surface modulated YIG films formed sharp band gaps. • A very small magnetic field change leads a large change in the peak value of band gap frequency. - Abstract: In this work, magnetostatic surface spin waves (MSSW) were propagated along the single crystal YIG (Y_3Fe_5O_1_2) film grown on GGG substrate. In order to obtain magnonic crystals, unlike the conventional methods, the surface of YIG films were magnetically modulated by magnet array in one and two-dimensions. The surface modulated YIG films formed sharp band gaps at approximately 6.55 GHz and 6.58 GHz at 1600 Oe magnetic field for one and two-dimensional magnonic crystals, respectively. It was found that a very small magnetic field change leads a large change in the peak value of band gap frequency.

  16. Surface modification of YIG by magnet array

    Energy Technology Data Exchange (ETDEWEB)

    Atalay, S., E-mail: satalay@inonu.edu.tr [Inonu University, Science and Art Faculty, Physics Department, 44280 Malatya (Turkey); Kolat, V.S. [Inonu University, Science and Art Faculty, Physics Department, 44280 Malatya (Turkey); Bakır, H.G. [Inonu University, Science and Art Faculty, Astronomy Department, 44280 Malatya (Turkey); Izgi, T.; Kaya, A.O. [Inonu University, Science and Art Faculty, Physics Department, 44280 Malatya (Turkey); Kaya, O.A. [Inonu University, Education Faculty, Computer Education and Educational Technology Department, 44280 Malatya (Turkey); Gencer, H. [Inonu University, Science and Art Faculty, Physics Department, 44280 Malatya (Turkey)

    2015-11-01

    Highlights: • The surface of YIG films were magnetically modulated by magnet array. • The surface modulated YIG films formed sharp band gaps. • A very small magnetic field change leads a large change in the peak value of band gap frequency. - Abstract: In this work, magnetostatic surface spin waves (MSSW) were propagated along the single crystal YIG (Y{sub 3}Fe{sub 5}O{sub 12}) film grown on GGG substrate. In order to obtain magnonic crystals, unlike the conventional methods, the surface of YIG films were magnetically modulated by magnet array in one and two-dimensions. The surface modulated YIG films formed sharp band gaps at approximately 6.55 GHz and 6.58 GHz at 1600 Oe magnetic field for one and two-dimensional magnonic crystals, respectively. It was found that a very small magnetic field change leads a large change in the peak value of band gap frequency.

  17. Preparation and Faraday rotation of Bi-YIG/PMMA nanocomposite

    Science.gov (United States)

    Fu, H. P.; Hong, R. Y.; Wu, Y. J.; Di, G. Q.; Xu, B.; Zheng, Y.; Wei, D. G.

    Bismuth-substituted yttrium iron garnet (Bi-YIG) nanoparticles (NPs) were prepared by coprecipitation and subsequent heating treatment. Thermal gravity-differential thermal analysis was performed to investigate the thermal behavior of the Bi-YIG precursors and to decide the best annealing temperature. Phase formation of garnet NPs was investigated by X-ray powder diffraction. The size of Bi-YIG NPs was investigated by transmission electron microscopy, and the magnetic properties of Bi-YIG NPs were measured using a vibrating sample magnetometer. The results show that the temperature needed for the transformation of Bi-YIG from the amorphous phase to the garnet phase decreases with increasing Bi content, and Bi-YIG NPs with sizes of 28-78 nm are obtained after heating treatment at 650-1000 °C. The saturation magnetization of Bi-YIG NPs increases as the Bi content increases. Moreover, the Faraday rotation of polymethyl methacrylate (PMMA) slices doped with Bi-YIG NPs was investigated. The results indicate that the angle of Faraday rotation increases with increasing Bi content in PMMA composites, and the maximum value of the figure of merit is 1.46°, which is comparable to the value of a sputtered film. The Bi-YIG NPs-doped PMMA slices are new promising materials for magneto-optical devices.

  18. PI-PfuI and PI-PfuII, intein-coded homing endonucleases from Pyrococcus furiosus. II. Characterization Of the binding and cleavage abilities by site-directed mutagenesis.

    OpenAIRE

    Komori, K; Ichiyanagi, K; Morikawa, K; Ishino, Y

    1999-01-01

    PI- Pfu I and PI- Pfu II from Pyrococcus furiosus are homing endonucleases, as shown in the accompanying paper. These two endonucleases are produced by protein splicing from the precursor protein including ribonucleotide reductase (RNR). We show here that both enzymes specifically interact with their substrate DNA and distort the DNA strands by 73 degrees and 67 degrees, respectively. They have two copies of the amino acid sequence motif LAGLIDADG, which is present in the majority of homing e...

  19. Complex group-I introns in nuclear SSU rDNA of red and green algae: evidence of homing-endonuclease pseudogenes in the Bangiophyceae

    DEFF Research Database (Denmark)

    Haugen, P; Huss, V A; Nielsen, Henrik

    1999-01-01

    on the complementary strand. A comparison between related group-I introns in the Bangiophyceae revealed homing-endonuclease-like pseudogenes due to frame-shifts and deletions in Porphyra and Bangia. The Scenedesmus and Porphyra introns provide new insights into the evolution and possible novel functions of nuclear...

  20. Improved propagation margin in YIG coated LPE garnet films for bubble devices

    International Nuclear Information System (INIS)

    Hidaka, Y.; Yoshimi, K.; Hibiya, T.; Mikami, M.

    1975-01-01

    YIG thin layers grown on (Y,Eu,Yb) 3 (Fe,Ga) 5 O 12 LPE films were found to be very effective for improvement of bubble propagation margin as well as for hard bubble suppression. In the ion-implanted rare earth substituted Ga:YIG on (111) GGG with 8 micron bubble, T-bar propagation margin was diminished, because of stretching or oscillating of bubble along the patterns. A 600 A YIG thin layer with in-plane magnetization, grown by CVD at 1065 0 C, obviated these destructive shortcomings and guaranteed the minimum driving field for stable bubble propagation down to 10 Oe. This improvement can be attributed to the magnetostatic interaction between the YIG layer and the bubble supporting layer. (auth)

  1. The mechanisms of action of E. coli endonuclease III and T4 UV endonuclease (endonuclease V) at AP sites.

    OpenAIRE

    Kim, J; Linn, S

    1988-01-01

    Treatment of DNA containing AP sites with either T4 UV endonuclease or with E. coli endonuclease III followed by a human class II AP endonuclease releases a putative beta-elimination product. This result suggests that both the T4 endonuclease and E. coli endonuclease III class I AP endonucleases catalyze phosphodiester bond cleavage via a lyase- rather than a hydrolase mechanism. Indeed, we have not detected a class I AP endonuclease which hydrolytically catalyzes phosphodiester bond cleavage...

  2. Low temperature spark plasma sintering of YIG powders

    International Nuclear Information System (INIS)

    Fernandez-Garcia, L.; Suarez, M.; Menendez, J.L.

    2010-01-01

    A transition from a low to a high spin state in the magnetization saturation between 1000 and 1100 o C calcination temperature is observed in YIG powders prepared by oxides mixture. Spark plasma sintering of these powders between 900 and 950 o C leads to dense samples with minimal formation of YFeO 3 , opening the way to co-sintering of YIG with metals or metallic alloys. The optical properties depend on the sintering stage: low (high) density samples show poor (bulk) optical absorption.

  3. Germline excision of transgenes in Aedes aegypti by homing endonucleases.

    Science.gov (United States)

    Aryan, Azadeh; Anderson, Michelle A E; Myles, Kevin M; Adelman, Zach N

    2013-01-01

    Aedes (Ae.) aegypti is the primary vector for dengue viruses (serotypes1-4) and chikungunya virus. Homing endonucleases (HEs) are ancient selfish elements that catalyze double-stranded DNA breaks (DSB) in a highly specific manner. In this report, we show that the HEs Y2-I-AniI, I-CreI and I-SceI are all capable of catalyzing the excision of genomic segments from the Ae. aegypti genome in a heritable manner. Y2-I-AniI demonstrated the highest efficiency at two independent genomic targets, with 20-40% of Y2-I-AniI-treated individuals producing offspring that had lost the target transgene. HE-induced DSBs were found to be repaired via the single-strand annealing (SSA) and non-homologous end-joining (NHEJ) pathways in a manner dependent on the availability of direct repeat sequences in the transgene. These results support the development of HE-based gene editing and gene drive strategies in Ae. aegypti, and confirm the utility of HEs in the manipulation and modification of transgenes in this important vector.

  4. Enhanced room-temperature spin Seebeck effect in a YIG/C60/Pt layered heterostructure

    Science.gov (United States)

    Das, R.; Kalappattil, V.; Geng, R.; Luong, H.; Pham, M.; Nguyen, T.; Liu, Tao; Wu, Mingzhong; Phan, M. H.; Srikanth, H.

    2018-05-01

    We report on large enhancement of the longitudinal spin Seebeck effect (LSSE) in the Y3Fe5O12 (YIG)/Pt system at room temperature due to the addition of a thin layer of organic semiconductor (C60) in between the YIG and the Pt. LSSE measurements show that the LSSE voltage increases significantly, from the initial value of 150 nV for the YIG/Pt structure to 240 nV for the YIG/C60(5nm)/Pt structure. Radio-frequency transverse susceptibility experiments reveal a significant decrease in the surface perpendicular magnetic anisotropy (PMA) of the YIG film when C60 is deposited on it. These results suggest that the LSSE enhancement may be attributed to increased spin mixing conductance, the decreased PMA, and the large spin diffusion length of C60.

  5. Visualizing phosphodiester-bond hydrolysis by an endonuclease

    DEFF Research Database (Denmark)

    Molina, Rafael; Stella, Stefano; Redondo, Pilar

    2015-01-01

    The enzymatic hydrolysis of DNA phosphodiester bonds has been widely studied, but the chemical reaction has not yet been observed. Here we follow the generation of a DNA double-strand break (DSB) by the Desulfurococcus mobilis homing endonuclease I-DmoI, trapping sequential stages of a two-metal-...

  6. Unidirectional Spin-Wave-Propagation-Induced Seebeck Voltage in a PEDOT:PSS/YIG Bilayer

    Science.gov (United States)

    Wang, P.; Zhou, L. F.; Jiang, S. W.; Luan, Z. Z.; Shu, D. J.; Ding, H. F.; Wu, D.

    2018-01-01

    We clarify the physical origin of the dc voltage generation in a bilayer of a conducting polymer film and a micrometer-thick magnetic insulator Y3Fe5O12 (YIG) film under ferromagnetic resonance and/or spin wave excitation conditions. The previous attributed mechanism, the inverse spin Hall effect in the polymer [Nat. Mater. 12, 622 (2013), 10.1038/nmat3634], is excluded by two control experiments. We find an in-plane temperature gradient in YIG which has the same angular dependence with the generated voltage. Both vanish when the YIG thickness is reduced to a few nanometers. Thus, we argue that the dc voltage is governed by the Seebeck effect in the polymer, where the temperature gradient is created by the nonreciprocal magnetostatic surface spin wave propagation in YIG.

  7. Whole genome sequencing of Rhodotorula mucilaginosa isolated from the chewing stick (Distemonanthus benthamianus): insights into Rhodotorula phylogeny, mitogenome dynamics and carotenoid biosynthesis.

    Science.gov (United States)

    Gan, Han Ming; Thomas, Bolaji N; Cavanaugh, Nicole T; Morales, Grace H; Mayers, Ashley N; Savka, Michael A; Hudson, André O

    2017-01-01

    In industry, the yeast Rhodotorula mucilaginosa is commonly used for the production of carotenoids. The production of carotenoids is important because they are used as natural colorants in food and some carotenoids are precursors of retinol (vitamin A). However, the identification and molecular characterization of the carotenoid pathway/s in species belonging to the genus Rhodotorula is scarce due to the lack of genomic information thus potentially impeding effective metabolic engineering of these yeast strains for improved carotenoid production. In this study, we report the isolation, identification, characterization and the whole nuclear genome and mitogenome sequence of the endophyte R. mucilaginosa RIT389 isolated from Distemonanthus benthamianus, a plant known for its anti-fungal and antibacterial properties and commonly used as chewing sticks. The assembled genome of R. mucilaginosa RIT389 is 19 Mbp in length with an estimated genomic heterozygosity of 9.29%. Whole genome phylogeny supports the species designation of strain RIT389 within the genus in addition to supporting the monophyly of the currently sequenced Rhodotorula species. Further, we report for the first time, the recovery of the complete mitochondrial genome of R. mucilaginosa using the genome skimming approach. The assembled mitogenome is at least 7,000 bases larger than that of Rhodotorula taiwanensis which is largely attributed to the presence of large intronic regions containing open reading frames coding for homing endonuclease from the LAGLIDADG and GIY-YIG families. Furthermore, genomic regions containing the key genes for carotenoid production were identified in R. mucilaginosa RIT389, revealing differences in gene synteny that may play a role in the regulation of the biotechnologically important carotenoid synthesis pathways in yeasts.

  8. Whole genome sequencing of Rhodotorula mucilaginosa isolated from the chewing stick (Distemonanthus benthamianus): insights into Rhodotorula phylogeny, mitogenome dynamics and carotenoid biosynthesis

    Science.gov (United States)

    Thomas, Bolaji N.; Cavanaugh, Nicole T.; Morales, Grace H.; Mayers, Ashley N.; Savka, Michael A.

    2017-01-01

    In industry, the yeast Rhodotorula mucilaginosa is commonly used for the production of carotenoids. The production of carotenoids is important because they are used as natural colorants in food and some carotenoids are precursors of retinol (vitamin A). However, the identification and molecular characterization of the carotenoid pathway/s in species belonging to the genus Rhodotorula is scarce due to the lack of genomic information thus potentially impeding effective metabolic engineering of these yeast strains for improved carotenoid production. In this study, we report the isolation, identification, characterization and the whole nuclear genome and mitogenome sequence of the endophyte R. mucilaginosa RIT389 isolated from Distemonanthus benthamianus, a plant known for its anti-fungal and antibacterial properties and commonly used as chewing sticks. The assembled genome of R. mucilaginosa RIT389 is 19 Mbp in length with an estimated genomic heterozygosity of 9.29%. Whole genome phylogeny supports the species designation of strain RIT389 within the genus in addition to supporting the monophyly of the currently sequenced Rhodotorula species. Further, we report for the first time, the recovery of the complete mitochondrial genome of R. mucilaginosa using the genome skimming approach. The assembled mitogenome is at least 7,000 bases larger than that of Rhodotorula taiwanensis which is largely attributed to the presence of large intronic regions containing open reading frames coding for homing endonuclease from the LAGLIDADG and GIY-YIG families. Furthermore, genomic regions containing the key genes for carotenoid production were identified in R. mucilaginosa RIT389, revealing differences in gene synteny that may play a role in the regulation of the biotechnologically important carotenoid synthesis pathways in yeasts. PMID:29158974

  9. Magnetic and structural properties of ZNn-doped YIG nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Garcia, R.P.; Del Toro, A.D.; Davila, Y.G.; Gomes, J.L.; Goncalves, L.P.; Hernandez, E.P. [Universidade Federal de Pernambuco (UFPE), PR (Brazil)

    2016-07-01

    Full text: Zn-doped YIG was prepared using the sol gel method and TGA measurements showed the phase formation between 900°C and 1000°C. XRD analysis showed close to 1100°C the formation of Franklinite phase, coexisting with the cubic YIG. Y3(Fe1-xZnx)5O12 samples with different Zn concentrations (x = 0, 0.01, 0.03 and 0.05) were prepared and analyzed by a magnetic study. A decrease in magnetic moment of the samples was confirmed with increasing the concentration of Zn ions. The diminishing is due to the substitution of Fe ions by Zn. This also confirmed the results of XRD showing the linear increasing in the lattice parameter. Fittings by the Bloch's law shows results compared with those already reported in the literature. Both the exponent of how Bloch constant presented similar values to those reported in YIG doped with other ions. We obtained the parameter α ∼ 10-6 K-n and n close to 1.9 for all samples. (author)

  10. Observation of spin superfluidity: YIG magnetic films and beyond

    Science.gov (United States)

    Sonin, Edouard

    2018-03-01

    From topology of the order parameter of the magnon condensate observed in yttrium-iron-garnet (YIG) magnetic films one must not expect energetic barriers making spin supercurrents metastable. But we show that some barriers of dynamical origin are possible nevertheless until the gradient of the phase (angle of spin precession) does not exceed the critical value (analog of the Landau critical velocity in superfluids). On the other hand, recently published claims of experimental detection of spin superfluidity in YIG films and antiferromagnets are not justified, and spin superfluidity in magnetically ordered solids has not yet been experimentally confirmed.

  11. Structural and magnetic properties of yttrium iron garnet (YIG) and yttrium aluminum iron garnet (YAIG) nanoferrites prepared by microemulsion method

    Energy Technology Data Exchange (ETDEWEB)

    Akhtar, Majid Niaz, E-mail: majidniazakhtar@ciitlahore.edu.pk [Department of Physics, COMSATS Institute of Information Technology, Lahore 54000 (Pakistan); Department of Mechanical and Materials Engineering, Faculty of Engineering and Built Environment, Universiti Kebangsaan Malaysia, 43600 Bangi, Selangor (Malaysia); Bakar Sulong, Abu [Department of Mechanical and Materials Engineering, Faculty of Engineering and Built Environment, Universiti Kebangsaan Malaysia, 43600 Bangi, Selangor (Malaysia); Khan, Muhammad Azhar [Department of Physics, The Islamia University of Bahawalpur, Bahawalpur 63100 (Pakistan); Ahmad, Mukhtar [Department of Physics, COMSATS Institute of Information Technology, Islamabad (Pakistan); Murtaza, Ghulam [Centre for Advanced Studies in Physics, G.C. University, Lahore, Pakistan" f Department of Mechanical Engineering, COMSATS Institute of Information Technology Sahiwal Pakistan (Pakistan); Raza, M.R. [Department of Mechanical and Materials Engineering, Faculty of Engineering and Built Environment, Universiti Kebangsaan Malaysia, 43600 Bangi, Selangor (Malaysia); Department of Mechanical Engineering, COMSATS Institute of Information Technology Sahiwal (Pakistan); Raza, R.; Saleem, M. [Department of Physics, COMSATS Institute of Information Technology, Lahore 54000 (Pakistan); Kashif, M. [Department of Physics, Govt. College University Faisalabad (Pakistan)

    2016-03-01

    Yttrium iron garnet (YIG) and yttrium aluminum iron garnet (YAIG) nanoferrite samples were synthesized by microemulsion method. The effect of sintering was examined by heating the samples at 900, 1000, and 1100 °C. The YIG and YAIG samples were then characterized using X-ray diffraction and field-emission scanning electron microscopy. Static and dynamic magnetic properties were measured by evaluating initial permeability, Q factor, and vibrating sample magnetometry properties of YIG and YAIG samples. YIG samples sintered at 1100 °C showed higher initial permeability and Q factor compared with YAIG samples. However, hysteresis loops also showed variations in the saturation magnetization, remanence, and coercivity of YIG and YAIG samples sintered at 900, 1000, and 1100 °C. The observed magnetic parameter such as saturation magnetization, coercivity and initial permeability are strongly affected by increasing temperature. The saturation magnetization and coercivity of YIG and YAIG nanoferrites were found in the range 11.56–19.92 emu/g and 7.30–87.70 Oe respectively. Furthermore, the decreasing trends in the static and magnetic properties of YAIG samples may be due to the introduction of Al ions in the YIG crystal lattice. Thus, YIG and YAIG sintered at 1100 °C can be used for wide-ranging frequency applications. - Highlights: • Static and dynamic magnetic properties of YIG and YAIG nanoferrites were determined. • Saturation magnetization, Q and initial permeability increased in YIG nanoferites. • Possible use of these nanoferrites for sensing and switching applications.

  12. YIG based broad band microwave absorber: A perspective on synthesis methods

    Science.gov (United States)

    Sharma, Vinay; Saha, J.; Patnaik, S.; Kuanr, Bijoy K.

    2017-10-01

    The fabrication of a thin layer of microwave absorber that operates over a wide band of frequencies is still a challenging task. With recent advances in nanostructure synthesis techniques, considerable progress has been achieved in realizations of thin nanocomposite layer designed for full absorption of incident electromagnetic (EM) radiation covering S to K band frequencies. The primary objective of this investigation is to achieve best possible EM absorption with a wide bandwidth and attenuation >10 dB for a thin absorbing layer (few hundred of microns). Magnetic yttrium iron garnet (Y3Fe5O12; in short YIG) nanoparticles (NPs) were prepared by sol-gel (SG) as well as solid-state (SS) reaction methods to elucidate the effects of nanoscale finite size on the magnetic behavior of the particles and hence their microwave absorption capabilities. It is found that YIG prepared by these two methods are different in many ways. Magnetic properties investigated using vibrating sample magnetometry (VSM) exhibit that the coercivity (Hc) of solid-state NPs is much larger (72 Oe) than the sol-gel NPs (31 Oe). Microwave absorption properties were studied by ferromagnetic resonance (FMR) technique in field sweep mode at different fixed frequencies. A thin layer (∼300 μm) of YIG film was deposited using electrophoretic deposition (EPD) technique over a coplanar waveguide (CPW) transmission line made on copper coated RT/duroid® 5880 substrates. Temperature dependent magnetic properties were also investigated using VSM and FMR techniques. Microwave absorption properties were investigated at high temperatures (up to 300 °C) both for sol-gel and solid-state synthesized NPs and are related to skin depth of YIG films. It is observed that microwave absorption almost vanishes when the temperature reached the Néel temperature of YIG.

  13. Effect of synthesis conditions on the preparation of YIG powders via co-precipitation method

    International Nuclear Information System (INIS)

    Rashad, M.M.; Hessien, M.M.; El-Midany, A.; Ibrahim, I.A.

    2009-01-01

    Yttrium iron garnet (YIG) (Y 3 Fe 5 O 12 ) powders have been synthesized through a co-precipitation method in the presence of sodium bis(2-ethylhexylsulfosuccinate), AOT as an anionic surfactant. The garnet precursors produced were obtained from aqueous iron and yttrium nitrates mixtures using 5 M sodium hydroxide at pH 10. A statistical Box-Behnken experimental design was used to investigate the effect of the main parameters (i.e. AOT surfactant concentration, annealing time and temperature) on YIG powder formation, crystallite size, morphology and magnetic properties. YIG particles were investigated using X-ray diffraction (XRD), scanning electron microscopy (SEM), and vibrating sample magnetometer. XRD revealed that the formation of single cubic phase of YIG was temperature dependent and increased by increasing the annealing temperature from 800 to 1200 o C. SEM micrographs showed that the addition of AOT surfactant promoted the microstructure of YIG in crystalline cubic-like structure. The magnetic properties were sensitive to the synthesis variables of annealing temperature, time and AOT surfactant concentration. The maximum saturation magnetization (28.13 emu/g), remanence magnetization (21.57 emu/g) and coercive force (703 Oe) were achieved at an annealing temperature of 1200 o C, time 2 h and 500 ppm of AOT surfactant concentration.

  14. The mitochondrial LSU rRNA group II intron of Ustilago maydis encodes an active homing endonuclease likely involved in intron mobility.

    Directory of Open Access Journals (Sweden)

    Anja Pfeifer

    Full Text Available BACKGROUND: The a2 mating type locus gene lga2 is critical for uniparental mitochondrial DNA inheritance during sexual development of Ustilago maydis. Specifically, the absence of lga2 results in biparental inheritance, along with efficient transfer of intronic regions in the large subunit rRNA gene between parental molecules. However, the underlying role of the predicted LAGLIDADG homing endonuclease gene I-UmaI located within the group II intron LRII1 has remained unresolved. METHODOLOGY/PRINCIPAL FINDINGS: We have investigated the enzymatic activity of I-UmaI in vitro based on expression of a tagged full-length and a naturally occurring mutant derivative, which harbors only the N-terminal LAGLIDADG domain. This confirmed Mg²⁺-dependent endonuclease activity and cleavage at the LRII1 insertion site to generate four base pair extensions with 3' overhangs. Specifically, I-UmaI recognizes an asymmetric DNA sequence with a minimum length of 14 base pairs (5'-GACGGGAAGACCCT-3' and tolerates subtle base pair substitutions within the homing site. Enzymatic analysis of the mutant variant indicated a correlation between the activity in vitro and intron homing. Bioinformatic analyses revealed that putatively functional or former functional I-UmaI homologs are confined to a few members within the Ustilaginales and Agaricales, including the phylogenetically distant species Lentinula edodes, and are linked to group II introns inserted into homologous positions in the LSU rDNA. CONCLUSIONS/SIGNIFICANCE: The present data provide strong evidence that intron homing efficiently operates under conditions of biparental inheritance in U. maydis. Conversely, uniparental inheritance may be critical to restrict the transmission of mobile introns. Bioinformatic analyses suggest that I-UmaI-associated introns have been acquired independently in distant taxa and are more widespread than anticipated from available genomic data.

  15. Selective inhibition by harmane of the apurinic apyrimidinic endonuclease activity of phage T4-induced UV endonuclease.

    Science.gov (United States)

    Warner, H R; Persson, M L; Bensen, R J; Mosbaugh, D W; Linn, S

    1981-11-25

    1-Methyl-9H-pyrido-[3,4-b]indole (harmane) inhibits the apurinic/apyrimidinic (AP) endonuclease activity of the UV endonuclease induced by phage T4, whereas it stimulates the pyrimidine dimer-DNA glycosylase activity of that enzyme. E. coli endonuclease IV, E. coli endonuclease VI (the AP endonuclease activity associated with E. coli exonuclease III), and E. coli uracil-DNA glycosylase were not inhibited by harmane. Human fibroblast AP endonucleases I and II also were only slightly inhibited. Therefore, harmane is neither a general inhibitor of AP endonucleases, nor a general inhibitor of Class I AP endonucleases which incise DNA on the 3'-side of AP sites. However, E. coli endonuclease III and its associated dihydroxythymine-DNA glycosylase activity were both inhibited by harmane. This observation suggests that harmane may inhibit only AP endonucleases which have associated glycosylase activities.

  16. Experimental investigation of the nature of the magnetoresistance effects in Pd-YIG hybrid structures.

    Science.gov (United States)

    Lin, Tao; Tang, Chi; Alyahayaei, Hamad M; Shi, Jing

    2014-07-18

    In bilayers consisting of Pd and yttrium iron garnet (Y(3)Fe(5)O(12) or YIG), we observe vanishingly small room-temperature conventional anisotropic magnetoresistance but large new magnetoresistance that is similar to the spin Hall magnetoresistance previously reported in Pt-YIG bilayers. We report a temperature dependence study of the two magnetoresistance effects in Pt-YIG bilayers. As the temperature is decreased, the new magnetoresistance shows a peak, whereas the anisotropic magnetoresistance effect starts to appear and increases monotonically. We find that the magnetoresistance peak shifts to lower temperatures in thicker Pd samples, a feature characteristic of the spin current effect. The distinct temperature dependence reveals fundamentally different mechanisms responsible for the two effects in such hybrid structures.

  17. Optimising homing endonuclease gene drive performance in a semi-refractory species: the Drosophila melanogaster experience.

    Directory of Open Access Journals (Sweden)

    Yuk-Sang Chan

    Full Text Available Homing endonuclease gene (HEG drive is a promising insect population control technique that employs meganucleases to impair the fitness of pest populations. Our previous studies showed that HEG drive was more difficult to achieve in Drosophila melanogaster than Anopheles gambiae and we therefore investigated ways of improving homing performance in Drosophila. We show that homing in Drosophila responds to increased expression of HEGs specifically during the spermatogonia stage and this could be achieved through improved construct design. We found that 3'-UTR choice was important to maximise expression levels, with HEG activity increasing as we employed Hsp70, SV40, vasa and βTub56D derived UTRs. We also searched for spermatogonium-specific promoters and found that the Rcd-1r promoter was able to drive specific expression at this stage. Since Rcd-1 is a regulator of differentiation in other species, it suggests that Rcd-1r may serve a similar role during spermatogonial differentiation in Drosophila. Contrary to expectations, a fragment containing the entire region between the TBPH gene and the bgcn translational start drove strong HEG expression only during late spermatogenesis rather than in the germline stem cells and spermatogonia as expected. We also observed that the fraction of targets undergoing homing was temperature-sensitive, falling nearly four-fold when the temperature was lowered to 18°C. Taken together, this study demonstrates how a few simple measures can lead to substantial improvements in the HEG-based gene drive strategy and reinforce the idea that the HEG approach may be widely applicable to a variety of insect control programs.

  18. A site-specific endonuclease encoded by a typical archaeal intron

    DEFF Research Database (Denmark)

    Dalgaard, Jacob; Garrett, Roger Antony; Belfort, Malene

    1993-01-01

    The protein encoded by the archaeal intron in the 23S rRNA gene of the hyperthermophile Desulfurococcus mobilis is a double-strand DNase that, like group I intron homing endonucleases, is capable of cleaving an intronless allele of the gene. This enzyme, I-Dmo I, is unusual among the intron...

  19. Preparation and microstructure of hot-pressed YIG

    International Nuclear Information System (INIS)

    Abdul Halim Shaari; Mansor Hashim; Sidek Haji Abd Aziz

    1992-01-01

    Bulk magnetic ceramics such as yttrium-iron-garnet (YIG) and nickel-zinc ferrite were prepared by the conventional-firing and hot-press techniques. Pre-sintering and sintering, both in air, were carried out at temperature ≥ 1200 0 C. Initial density and microstructural measurements show that the high density samples with uniform grain size can be readily achieved by hot-pressing. The well-defined microstructures, the high density attained. The agreement of saturation magnetization 4πMs values and the high resistivities indicated that the optimized preparation conditions for conventional firing yielded yttrium-iron-garnet of good quality. Quenched and isochronally annealed, the YIG samples were also found to retain their room-temperature saturation-magnetization and resonance linewidth values after any single heat treatment. However, the electrical resistivity and the effective linewidth clearly exhibited isochronal recovery. A possible model to explain this behaviour and its implications on microwave applications are discussed. Ferrites of the type MFe 2 O 4 where M=Mg, Mn, Co, Fe, Cu, Zn and Cd and of the type Ni1-2xSnxFe 2 O4 have also been prepared and their properties are being studied

  20. ORIGIN FOR IRRADIATION EFFECT OF 0.56GeV C6+ ON CaVSn:YIG

    Institute of Scientific and Technical Information of China (English)

    熊宏齐; 侯明东; 等

    1995-01-01

    This paper presents numerous physical characteristics of Ca,V.Sn doped yttrium iron garnet(CaVSn:YIG) irradiated with 0.56GeV carbon ions delivered by the Heavy Ion Research Facility of Lanzhou (HIRFL).The reason for change of the magnetic properties of the samples induced by energetic carbon ions bombardment is discussed.By comparison of this results with the irradiation effects of YIG induced by eneregetic argon,krypton and xenon oibtained on the GANIL,Caen,France,it is concluded that the irradiation effect of 0.56GeV C6+ on CaVSn:YIG arises from the electronic energy losses.

  1. Effect of NiO inserted layer on spin-Hall magnetoresistance in Pt/NiO/YIG heterostructures

    International Nuclear Information System (INIS)

    Shang, T.; Zhan, Q. F.; Yang, H. L.; Zuo, Z. H.; Xie, Y. L.; Liu, L. P.; Zhang, S. L.; Zhang, Y.; Li, H. H.; Wang, B. M.; Li, Run-Wei; Wu, Y. H.; Zhang, S.

    2016-01-01

    We investigate spin-current transport with an antiferromagnetic insulator NiO thin layer by means of the spin-Hall magnetoresistance (SMR) over a wide range of temperature in Pt/NiO/Y_3Fe_5O_1_2 (Pt/NiO/YIG) heterostructures. The SMR signal is comparable to that without the NiO layer as long as the temperature is near or above the blocking temperature of the NiO, indicating that the magnetic fluctuation of the insulating NiO is essential for transmitting the spin current from the Pt to YIG layer. On the other hand, the SMR signal becomes negligibly small at low temperature, and both conventional anisotropic magnetoresistance and the anomalous Hall resistance are extremely small at any temperature, implying that the insertion of the NiO has completely suppressed the Pt magnetization induced by the YIG magnetic proximity effect (MPE). The dual roles of the thin NiO layer are, to suppress the magnetic interaction or MPE between Pt and YIG, and to maintain efficient spin current transmission at high temperature.

  2. Influence of static and dynamic dipolar fields in bulk YIG/thin film NiFe systems probed via spin rectification effect

    Energy Technology Data Exchange (ETDEWEB)

    Soh, Wee Tee, E-mail: a0046479@u.nus.edu [Center for Superconducting and Magnetic Materials, Department of Physics, National University of Singapore, 2 Science Drive 3, Singapore 117551 (Singapore); Tay, Z.J. [Center for Superconducting and Magnetic Materials, Department of Physics, National University of Singapore, 2 Science Drive 3, Singapore 117551 (Singapore); Yakovlev, N.L. [Institute of Materials Research and Engineering, A*STAR (Agency for Science, Technology and Research), 3 Research Link, Singapore 117602 (Singapore); Peng, Bin [State Key Laboratory of Electronic Thin Films and Integrated Devices, University of Electronic Science and Technology of China, Chengdu 610054 (China); Ong, C.K. [Center for Superconducting and Magnetic Materials, Department of Physics, National University of Singapore, 2 Science Drive 3, Singapore 117551 (Singapore); Temasek Laboratories, National University of Singapore, 5A Engineering Drive 2, Singapore 117411 (Singapore)

    2017-03-15

    The characteristics of the static and dynamic components of the dipolar fields originating from a bulk polycrystalline yttrium iron garnet (YIG) substrate are probed by depositing a NiFe (Permalloy) layer on it, which acts as a detector. By measuring dc voltages generated via spin rectification effect (SRE) within the NiFe layer under microwave excitation, we characterize the influence of dipolar fields from bulk YIG on the NiFe layer. It is found that the dynamic YIG dipolar fields modify the self-SRE of NiFe, driving its own rectification voltages within the NiFe layer, an effect we term as non-local SRE. This non-local SRE only occurs near the simultaneous resonance of both YIG and NiFe. On the other hand, the static dipolar field from YIG manifests itself as a negative anisotropy in the NiFe layer which shifts the latter’s ferromagnetic resonance frequency. - Highlights: • We demonstrate the quantification of both the static and dynamic components of the dipolar fields due to a YIG slab. • The detection and characterisation of such dipolar fields are important in many magnetic applications such as magnonics. • The dipolar fields can pose potential pitfalls if not properly considered in certain spin-electronics systems.

  3. Spin-Orbit Torque and Spin Pumping in YIG/Pt with Interfacial Insertion Layers (Postprint)

    Science.gov (United States)

    2018-05-03

    modified by spin-orbit torque6,7 in thin- film YIG due to absorption of pure spin current,8–12 which is gen- erated from an electric current in the adjacent... films were grown on Gd3Ga5O12(111) substrates by pulsed laser deposition as reported in Ref. 3. The YIG films were transferred through an ambient... introduction into the deposition chamber, maintained at 250 C at 50 mTorr O2 for 30 min to remove water and organics on the surface. The metal overlayers

  4. Study of photon–magnon coupling in a YIG-film split-ring resonant system

    Energy Technology Data Exchange (ETDEWEB)

    Bhoi, B.; Aiyar, R. [Department of Physics, Indian Institute of Technology Bombay, Powai, Mumbai 400076 (India); CRNTS, Indian Institute of Technology Bombay, Powai, Mumbai 400076 (India); Cliff, T.; Maksymov, I. S.; Kostylev, M., E-mail: mikhail.kostylev@uwa.edu.au [School of Physics M013, University of Western Australia, Crawley 6009 (Australia); Venkataramani, N. [Department of Metallurgical Engineering and Materials Science, Indian Institute of Technology Bombay, Mumbai 400076 (India); Prasad, S. [Department of Physics, Indian Institute of Technology Bombay, Powai, Mumbai 400076 (India); Stamps, R. L. [School of Physics M013, University of Western Australia, Crawley 6009 (Australia); SUPA, University of Glasgow, Glasgow G12 8QQ (United Kingdom)

    2014-12-28

    By using the stripline Microwave Vector–Network Analyser Ferromagnetic Resonance and Time Domain spectroscopy techniques, we study a strong coupling regime of magnons to microwave photons in the planar geometry of a lithographically formed split-ring resonator (SRR) loaded by a single-crystal epitaxial yttrium–iron–garnet (YIG) film. Strong anti-crossing of the photon modes of SRR and of the magnon modes of the YIG film is observed in the applied-magnetic-field resolved measurements. The coupling strength extracted from the experimental data reaches 9% at 3 GHz. Theoretically, we propose an equivalent circuit model of the SRR loaded by a magnetic film. This model follows from the results of our numerical simulations of the microwave field structure of the SRR and of the magnetisation dynamics in the YIG film driven by the microwave currents in the SRR. The results obtained with the equivalent-circuit model are in good agreement with the experiment. This model provides a simple physical explanation of the process of mode anti-crossing. Our findings are important for future applications in microwave quantum photonic devices as well as in nonlinear and magnetically tuneable metamaterials exploiting the strong coupling of magnons to microwave photons.

  5. YIG: Bi2O3 Nanocomposite Thin Films for Magnetooptic and Microwave Applications

    Directory of Open Access Journals (Sweden)

    M. Nur-E-Alam

    2015-01-01

    Full Text Available Y3Fe5O12-Bi2O3 composite thin films are deposited onto Gd3Ga5O12 (GGG substrates and their annealing crystallization regimes are optimized (in terms of both process temperatures and durations to obtain high-quality thin film layers possessing magnetic properties attractive for a range of technological applications. The amount of bismuth oxide content introduced into these nanocomposite-type films is controlled by adjusting the RF power densities applied to both Y3Fe5O12 and Bi2O3 sputtering targets during the cosputtering deposition processes. The measured material properties of oven-annealed YIG-Bi2O3 films indicate that cosputtering of YIG-Bi2O3 composites can provide the flexibility of application-specific YIG layers fabrication of interest for several existing, emerging, and also frontier technologies. Experimental results demonstrate large specific Faraday rotation (of more than 1°/µm at 532 nm, achieved simultaneously with low optical losses in the visible range and very narrow peak-to-peak ferromagnetic resonance linewidth of around ΔHpp= 6.1 Oe at 9.77 GHz.

  6. Pure nuclear reflexes and combined hyperfine interactions in YIG

    Energy Technology Data Exchange (ETDEWEB)

    Winkler, H; Eisberg, R; Alp, E; Rueffer, R; Gerdau, E; Lauer, S; Trautwein, A X; Grodzicki, M; Vera, A

    1983-01-01

    Moessbauer spectra of oriented YIG single crystals were taken and the numerical analysis using the transmission integral yielded a consistent set of hyperfine interaction parameters. They are in good agreement with theoretical values obtained by MO-calculations which included clusters up to 62 ions. Finally pure nuclear reflexes are predicted for single crystals and two theoretical spectra are given.

  7. EsrE-A yigP Locus-Encoded Transcript-Is a 3′ UTR sRNA Involved in the Respiratory Chain of E. coli

    Directory of Open Access Journals (Sweden)

    Hui Xia

    2017-08-01

    Full Text Available The yigP locus is widely conserved among γ-proteobacteria. Mutation of the yigP locus impacts aerobic growth of Gram-negative bacteria. However, the underlying mechanism of how the yigP locus influences aerobic growth remains largely unknown. Here, we demonstrated that the yigP locus in Escherichia coli encodes two transcripts; the mRNA of ubiquinone biosynthesis protein, UbiJ, and the 3′ untranslated region small regulatory RNA (sRNA, EsrE. EsrE is an independent transcript that is transcribed using an internal promoter of the yigP locus. Surprisingly, we found that both the EsrE sRNA and UbiJ protein were required for Q8 biosynthesis, and were sufficient to rescue the growth defect ascribed to deletion of the yigP locus. Moreover, our data showed that EsrE targeted multiple mRNAs involved in several cellular processes including murein biosynthesis and the tricarboxylic acid cycle. Among these targets, sdhD mRNA that encodes one subunit of succinate dehydrogenase (SDH, was significantly activated. Our findings provided an insight into the important function of EsrE in bacterial adaptation to various environments, as well as coordinating different aspects of bacterial physiology.

  8. Structural and magnetic characterization of YIG particles prepared using microemulsions

    International Nuclear Information System (INIS)

    Teijeiro, A.G.; Baldomir, D.; Rivas, J.; Paz, S.; Vaqueiro, P.; Lopez Quintela, A.

    1995-01-01

    Yttrium-iron-garnet (YIG) particles have been synthesized using the microemulsion technique. A comparison of ferrite powders obtained by this method and those prepared by sol-gel and solid state reactions is reported. We have studied both the magnetic and structural properties and have found a dependence on annealing temperatures. ((orig.))

  9. Preparation of yttrium iron garnet (YIG) by modified domestic iron oxide

    International Nuclear Information System (INIS)

    Mozaffari, M.; Amighian

    2002-01-01

    Iron oxide by product of a local steel complex was modified to use for preparation of Yttrium iron garnet (YIG). The improvement was necessary to reduce impurities, especially the Si0 2 and Cl contents, which have deteriorative effects on magnetic properties and equipment used for preparation of the samples. The modified iron oxide was then mixed with Yttrium oxide of Merck Company in appropriate proportion to obtain a stoichiometric single phase YIG, using the conventional ceramic technique. XRD and SEM equipments were used to identify the resulting phases and microstructure respectively. Magnetic parameters were measured by VSM. Curie temperature of the samples was obtained by DTG (M) method. The results were compared with those obtained from samples that made by Merck iron oxide. There are small differences between the results. This was discussed according to extra pores and minute secondary phase in the samples made by domestic iron oxide. (Author)

  10. Magneto-optical properties of yttrium iron garnet (YIG) thin films elaborated by radio frequency sputtering

    International Nuclear Information System (INIS)

    Boudiar, T.; Payet-Gervy, B.; Blanc-Mignon, M.-F.; Rousseau, J.-J.; Le Berre, M.; Joisten, H.

    2004-01-01

    Thin films of yttrium iron garnet (YIG) are grown by radio frequency magnetron non reactive sputtering system. Thin films are crystallised by heat-treatment to obtain magneto-optical properties. On quartz substrate, the network of cracks observed on the annealed samples can be explained by the difference between the thermal expansion coefficient of substrate and YIG. Physico-chemical analysis shown that the obtained material has a correct stoichiometry and is crystallised as FCC. The Faraday rotation of thin films is measured with a classical ellipsometric system based on transmission which allows us to obtained an accuracy of 0.01 deg. The variation of Faraday rotation is studied on the one hand versus radio frequency power applied to the cathode during the deposition and on the other hand versus the applied magnetic field. The results are compared with those obtained by vibrating sample magnetometer analysis in perpendicular configuration. A maximum Faraday rotation is observed to be 1900 deg./cm at the wavelength of 594nm for a YIG thin film formed on quartz substrate and annealed at 740 deg. C. The values of the Faraday rotation coefficients obtained in the study versus the wavelength are comparable to those of the literature for the bulk material. In order to eliminate the stress due to the heat-treatment, we made some films on single crystals of gadolinium gallium garnet (111) substrates for which thermal expansion coefficient is near than the YIG one. The material crystallises with no crackles and the Faraday effect is equivalent

  11. Influence of Thickness and Interface on the Low-Temperature Enhancement of the Spin Seebeck Effect in YIG Films

    Directory of Open Access Journals (Sweden)

    Er-Jia Guo

    2016-07-01

    Full Text Available The temperature-dependent longitudinal spin Seebeck effect (LSSE in heavy metal (HM/Y_{3}Fe_{5}O_{12} (YIG hybrid structures is investigated as a function of YIG film thickness, magnetic field strength, and different HM detection materials. The LSSE signal shows a large enhancement with reductions in temperature, leading to a pronounced peak at low temperatures. We find that the LSSE peak temperature strongly depends on the film thickness as well as on the magnetic field. Our result can be well explained in the framework of magnon-driven LSSE by taking into account the temperature-dependent effective propagation length of thermally excited magnons in the bulk of the material. We further demonstrate that the LSSE peak is significantly shifted by changing the interface coupling to an adjacent detection layer, revealing a more complex behavior beyond the currently discussed bulk effect. By direct microscopic imaging of the interface, we correlate the observed temperature dependence with the interface structure between the YIG and the adjacent metal layer. Our results highlight the role of interface effects on the temperature-dependent LSSE in HM/YIG system, suggesting that the temperature-dependent spin current transparency strikingly relies on the interface conditions.

  12. Large enhancement of Faraday rotation by localized surface plasmon resonance in Au nanoparticles embedded in Bi:YIG film

    International Nuclear Information System (INIS)

    Uchida, H.; Masuda, Y.; Fujikawa, R.; Baryshev, A.V.; Inoue, M.

    2009-01-01

    A large enhancement of the Faraday rotation, which is associated with localized surface plasmon resonance (LSPR), was obtained in a sample with Au nanoparticles embedded in a Bi-substituted yttrium iron garnet (Bi:YIG) film. On a quartz substrate, Au nanoparticles were formed by heating an Au thin film, and a Bi:YIG film was then deposited on them. A sample containing the Au nanoparticles produced by 1000 deg. C heating showed a resonant attenuation with narrower bandwidth in the transmission spectrum than nanoparticles of other samples formed by low-temperature heating. The sharp resonant Faraday rotation angle was 4.4 times larger than the estimated intrinsic Bi:YIG film at the LSPR wavelength; the angular difference was 0.14 deg. A discrepancy in the bandwidth between the transmission attenuation and the resonant Faraday rotation is discussed

  13. Using Group II Introns for Attenuating the In Vitro and In Vivo Expression of a Homing Endonuclease.

    Directory of Open Access Journals (Sweden)

    Tuhin Kumar Guha

    Full Text Available In Chaetomium thermophilum (DSM 1495 within the mitochondrial DNA (mtDNA small ribosomal subunit (rns gene a group IIA1 intron interrupts an open reading frame (ORF encoded within a group I intron (mS1247. This arrangement offers the opportunity to examine if the nested group II intron could be utilized as a regulatory element for the expression of the homing endonuclease (HEase. Constructs were generated where the codon-optimized ORF was interrupted with either the native group IIA1 intron or a group IIB type intron. This study showed that the expression of the HEase (in vivo in Escherichia coli can be regulated by manipulating the splicing efficiency of the HEase ORF-embedded group II introns. Exogenous magnesium chloride (MgCl2 stimulated the expression of a functional HEase but the addition of cobalt chloride (CoCl2 to growth media antagonized the expression of HEase activity. Ultimately the ability to attenuate HEase activity might be useful in precision genome engineering, minimizing off target activities, or where pathways have to be altered during a specific growth phase.

  14. Endonuclease IV Is the major apurinic/apyrimidinic endonuclease in Mycobacterium tuberculosis and is important for protection against oxidative damage.

    Directory of Open Access Journals (Sweden)

    Rupangi Verma Puri

    Full Text Available During the establishment of an infection, bacterial pathogens encounter oxidative stress resulting in the production of DNA lesions. Majority of these lesions are repaired by base excision repair (BER pathway. Amongst these, abasic sites are the most frequent lesions in DNA. Class II apurinic/apyrimidinic (AP endonucleases play a major role in BER of damaged DNA comprising of abasic sites. Mycobacterium tuberculosis, a deadly pathogen, resides in the human macrophages and is continually subjected to oxidative assaults. We have characterized for the first time two AP endonucleases namely Endonuclease IV (End and Exonuclease III (XthA that perform distinct functions in M.tuberculosis. We demonstrate that M.tuberculosis End is a typical AP endonuclease while XthA is predominantly a 3'→5' exonuclease. The AP endonuclease activity of End and XthA was stimulated by Mg(2+ and Ca(2+ and displayed a preferential recognition for abasic site paired opposite to a cytosine residue in DNA. Moreover, End exhibited metal ion independent 3'→5' exonuclease activity while in the case of XthA this activity was metal ion dependent. We demonstrate that End is not only a more efficient AP endonuclease than XthA but it also represents the major AP endonuclease activity in M.tuberculosis and plays a crucial role in defense against oxidative stress.

  15. Study of the magnetic disaccommodation in La doped YIG

    International Nuclear Information System (INIS)

    Torres, C.; Hernandez-Gomez, P.; Francisco, C. de; Munoz, J.M.; Alejos, O.; Gonzalez Arias, A.; Perdigao, J.M.; Ferreira, A.R.

    2005-01-01

    The relaxation of the initial magnetic permeability of La doped yttrium iron garnet (YIG) samples with nominal composition Y 3-x La x Fe 5 O 12 (0 2 atmosphere when the La content is at least of 0.3. These results have been interpreted in terms of the formation of a secondary perovskite phase when the La solubility limit is reached. (copyright 2005 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim) (orig.)

  16. Expression of homing endonuclease gene and insertion-like element in sea anemone mitochondrial genomes: Lesson learned from Anemonia viridis.

    Science.gov (United States)

    Chi, Sylvia Ighem; Urbarova, Ilona; Johansen, Steinar D

    2018-04-30

    The mitochondrial genomes of sea anemones are dynamic in structure. Invasion by genetic elements, such as self-catalytic group I introns or insertion-like sequences, contribute to sea anemone mitochondrial genome expansion and complexity. By using next generation sequencing we investigated the complete mtDNAs and corresponding transcriptomes of the temperate sea anemone Anemonia viridis and its closer tropical relative Anemonia majano. Two versions of fused homing endonuclease gene (HEG) organization were observed among the Actiniidae sea anemones; in-frame gene fusion and pseudo-gene fusion. We provided support for the pseudo-gene fusion organization in Anemonia species, resulting in a repressed HEG from the COI-884 group I intron. orfA, a putative protein-coding gene with insertion-like features, was present in both Anemonia species. Interestingly, orfA and COI expression were significantly up-regulated upon long-term environmental stress corresponding to low seawater pH conditions. This study provides new insights to the dynamics of sea anemone mitochondrial genome structure and function. Copyright © 2018 Elsevier B.V. All rights reserved.

  17. Exchange magnetic field torques in YIG/Pt bilayers observed by the spin-Hall magnetoresistance

    NARCIS (Netherlands)

    Vlietstra, N.; Shan, J.; Castel, V.; Ben Youssef, J.; Bauer, G. E. W.; van Wees, B. J.

    2013-01-01

    The effective field torque of an yttrium-iron-garnet (YIG) film on the spin accumulation in an attached platinum (Pt) film is measured by the spin-Hall magnetoresistance (SMR). As a result, the magnetization direction of a ferromagnetic insulating layer can be measured electrically. Experimental

  18. Robust magnon-photon coupling in a planar-geometry hybrid of inverted split-ring resonator and YIG film.

    Science.gov (United States)

    Bhoi, Biswanath; Kim, Bosung; Kim, Junhoe; Cho, Young-Jun; Kim, Sang-Koog

    2017-09-20

    We experimentally demonstrate strongly enhanced coupling between excited magnons in an Yttrium Iron Garnet (YIG) film and microwave photons in an inverted pattern of split-ring resonator (noted as ISRR). The anti-crossing effects of the ISRR's photon mode and the YIG's magnon modes were found from |S 21 |-versus-frequency measurements for different strengths and directions of externally applied magnetic fields. The spin-number-normalized coupling strength (i.e. single spin-photon coupling) [Formula: see text] was determined to 0.194 Hz ([Formula: see text] = 90 MHz) at 3.7 GHz frequency. Furthermore, we found that additional fine features in the anti-crossing region originate from the excitation of different spin-wave modes (such as the magnetostatic surface and the backward-volume magnetostatic spin-waves) rather than the Kittel-type mode. These spin-wave modes, as coupled with the ISRR mode, modify the anti-crossing effect as well as their coupling strength. An equivalent circuit model very accurately reproduced the observed anti-crossing effect and its coupling strength variation with the magnetic field direction in the planar-geometry ISRR/YIG hybrid system. This work paves the way for the design of new types of high-gain magnon-photon coupling systems in planar geometry.

  19. Endonuclease activities in extracts of Micrococcus luteus that act on. gamma. -irradiated DNA

    Energy Technology Data Exchange (ETDEWEB)

    Schoen-Bopp, A; Schaefer, G; Hagen, U [Kernforschungszentrum Karlsruhe (Germany, F.R.). Inst. fuer Strahlenbiologie

    1977-03-01

    Several protein fractions containing endonuclease activity against ..gamma..-irradiated DNA (..gamma..-endonuclease) were isolated from M.luteus. The crude extract was eluted on a phosphocellulose column and chromatographed on TEAE cellulose and subsequently on hydroxypatite. Five peaks of ..gamma..-endonuclease were obtained from each preparation. Repeated experiments showed comparable chromatographic behaviour of the fractions. There was no detectable activity of uv-endonuclease in the fractions with ..gamma..-endonuclease but a small contamination of endonuclease against unirradiated DNA and against DNA with apurinic sites. The ..gamma..-endonuclease was stimulated by, but was not dependent on, magnesium. Several tests for endonuclease activity have been used: the analysis of strand breaks in calf-thymus DNA or in PM2 DNA, and the determination of end-groups formed by endonuclease, either 3'OH end-groups or phosphomonoester end groups. From the results obtained it can be assumed that the strand breaks induced by the ..gamma..-endonuclease carry 3'OH and 5' phosphate end groups.

  20. Genes ycfR, sirA and yigG contribute to the surface attachment of Salmonella enterica Typhimurium and Saintpaul to fresh produce.

    Directory of Open Access Journals (Sweden)

    Joelle K Salazar

    Full Text Available Salmonella enterica is a frequent contaminant of minimally-processed fresh produce linked to major foodborne disease outbreaks. The molecular mechanisms underlying the association of this enteric pathogen with fresh produce remain largely unexplored. In our recent study, we showed that the expression of a putative stress regulatory gene, ycfR, was significantly induced in S. enterica upon exposure to chlorine treatment, a common industrial practice for washing and decontaminating fresh produce during minimal processing. Two additional genes, sirA involved in S. enterica biofilm formation and yigG of unknown function, were also found to be differentially regulated under chlorine stress. To further characterize the roles of ycfR, sirA, and yigG in S. enterica attachment and survival on fresh produce, we constructed in-frame deletions of all three genes in two different S. enterica serovars, Typhimurium and Saintpaul, which have been implicated in previous disease outbreaks linked to fresh produce. Bacterial attachment to glass and polystyrene microtiter plates, cell aggregation and hydrophobicity, chlorine resistance, and surface attachment to intact spinach leaf and grape tomato were compared among wild-type strains, single-gene deletion mutants, and their respective complementation mutants. The results showed that deletions of ycfR, sirA, and yigG reduced bacterial attachment to glass and polystyrene as well as fresh produce surface with or without chlorine treatment in both Typhimurium and Saintpaul. Deletion of ycfR in Typhimurium significantly reduced bacterial chlorine resistance and the attachment to the plant surfaces after chlorinated water washes. Deletions of ycfR in Typhimurium and yigG in Saintpaul resulted in significant increase in cell aggregation. Our findings suggest that ycfR, sirA, and yigG collectively contribute to S. enterica surface attachment and survival during post-harvest minimal processing of fresh produce.

  1. A yigP mutant strain is a small colony variant of E. coli and shows pleiotropic antibiotic resistance.

    Science.gov (United States)

    Xia, Hui; Tang, Qiongwei; Song, Jie; Ye, Jiang; Wu, Haizhen; Zhang, Huizhan

    2017-12-01

    Small colony variants (SCVs) are a commonly observed subpopulation of bacteria that have a small colony size and distinctive biochemical characteristics. SCVs are more resistant than the wild type to some antibiotics and usually cause persistent infections in the clinic. SCV studies have been very active during the past 2 decades, especially Staphylococcus aureus SCVs. However, fewer studies on Escherichia coli SCVs exist, so we studied an E. coli SCV during an experiment involving the deletion of the yigP locus. PCR and DNA sequencing revealed that the SCV was attributable to a defect in the yigP function. Furthermore, we investigated the antibiotic resistance profile of the E. coli SCV and it showed increased erythromycin, kanamycin, and d-cycloserine resistance, but collateral sensitivity to ampicillin, polymyxin, chloramphenicol, tetracycline, rifampin, and nalidixic acid. We tried to determine the association between yigP and the pleiotropic antibiotic resistance of the SCV by analyzing biofilm formation, cellular morphology, and coenzyme Q (Q 8 ) production. Our results indicated that impaired Q 8 biosynthesis was the primary factor that contributed to the increased resistance and collateral sensitivity of the SCV. This study offers a novel genetic basis for E. coli SCVs and an insight into the development of alternative antimicrobial strategies for clinical therapy.

  2. Creating a monomeric endonuclease TALE-I-SceI with high specificity and low genotoxicity in human cells.

    Science.gov (United States)

    Lin, Jianfei; Chen, He; Luo, Ling; Lai, Yongrong; Xie, Wei; Kee, Kehkooi

    2015-01-01

    To correct a DNA mutation in the human genome for gene therapy, homology-directed repair (HDR) needs to be specific and have the lowest off-target effects to protect the human genome from deleterious mutations. Zinc finger nucleases, transcription activator-like effector nuclease (TALEN) and CRISPR-CAS9 systems have been engineered and used extensively to recognize and modify specific DNA sequences. Although TALEN and CRISPR/CAS9 could induce high levels of HDR in human cells, their genotoxicity was significantly higher. Here, we report the creation of a monomeric endonuclease that can recognize at least 33 bp by fusing the DNA-recognizing domain of TALEN (TALE) to a re-engineered homing endonuclease I-SceI. After sequentially re-engineering I-SceI to recognize 18 bp of the human β-globin sequence, the re-engineered I-SceI induced HDR in human cells. When the re-engineered I-SceI was fused to TALE (TALE-ISVB2), the chimeric endonuclease induced the same HDR rate at the human β-globin gene locus as that induced by TALEN, but significantly reduced genotoxicity. We further demonstrated that TALE-ISVB2 specifically targeted at the β-globin sequence in human hematopoietic stem cells. Therefore, this monomeric endonuclease has the potential to be used in therapeutic gene targeting in human cells. © The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.

  3. Key Players in I-DmoI Endonuclease Catalysis Revealed from Structure and Dynamics

    DEFF Research Database (Denmark)

    Molina, Rafael; Besker, Neva; Marcaida, Maria Jose

    2016-01-01

    . The cleavage mechanism was related both to key structural effects, such as the position of water molecules and ions participating in the cleavage reaction, and to dynamical effects related to protein behavior. In particular, we found that the protein perturbation pattern significantly changes between cleaved......Homing endonucleases, such as I-DmoI, specifically recognize and cleave long DNA target sequences (∼20 bp) and are potentially powerful tools for genome manipulation. However, inefficient and off-target DNA cleavage seriously limits specific editing in complex genomes. One approach to overcome...

  4. Crystal structure of the apurinic/apyrimidinic endonuclease IV from Mycobacterium tuberculosis.

    Science.gov (United States)

    Zhang, Wei; Xu, Yueyang; Yan, Mengrong; Li, Shanshan; Wang, Huiying; Yang, Haitao; Zhou, Weihong; Rao, Zihe

    2018-03-25

    Endonuclease IV is a typical endonuclease of the apurinic-apyrimidinic (AP) or abasic endonuclease superfamily. It repairs damaged DNA through base excision repair by cleaving the DNA backbone immediately 5' of an AP site. In Mycobacterium tuberculosis, endonuclease IV is the major AP endonuclease. This enzyme is absent from mammalian cells, making it an attractive target for anti-tuberculosis drug development. In this study, the structure of the recombinant endonuclease IV from M. tuberculosis (MtbEndo IV) was determined at a high resolution of 1.18 Å. MtbEndo IV was found to have a classical α8β8-fold TIM barrel with loops on its surface connecting the α-helices and β-strands that constitute a groove for DNA binding. Three zinc ions were identified at the active site. A comparison between the structures of MtbEndo IV and Escherichia coli End IV suggested that Gln32 of MtbEndo IV may plays a role in regulating substrate binding. Copyright © 2018. Published by Elsevier Inc.

  5. Homing at an extragenic locus mediated by VDE (PI-SceI) in Saccharomyces cerevisiae.

    Science.gov (United States)

    Nogami, Satoru; Fukuda, Tomoyuki; Nagai, Yuri; Yabe, Shizu; Sugiura, Masako; Mizutani, Ryuta; Satow, Yoshinori; Anraku, Yasuhiro; Ohya, Yoshikazu

    2002-06-30

    PI-SceI (VDE), a homing endonuclease with protein splicing activity, is a genomic parasite in the VMA1 gene of Saccharomyces cerevisiae. In a heterozygous diploid of the VDE-less VMA1 allele and a VDE-containing VMA1 allele, VDE specifically cleaves its recognition sequence (VRS) in the VDE-less VMA1 allele at meiosis, followed by 'homing', i.e. a conversion to a VDE-containing allele. We found that upon VDE expression, homing of a marker gene at an extragenic locus occurs only when a 45 bp element containing the VRS is inserted at its allelic site, while mutants of VDE with no endonuclease activity lack authentic extragenic homing activity. Thus, both the VRS and VDE are required for homing. Insertion of the VRS in a homozygous diploid significantly lowered the spore germination ability, indicating that a template for gene repair at its allelic locus is essential for efficient homing and survival of yeast cells. Copyright 2002 John Wiley & Sons, Ltd.

  6. Endonuclease IV of Escherichia coli is induced by paraquat

    Energy Technology Data Exchange (ETDEWEB)

    Chan, E.; Weiss, B.

    1987-05-01

    The addition of paraquat (methyl viologen) to a growing culture of Escherichia coli K-12 led within 1 hr to a 10- to 20-fold increase in the level of endonuclease IV, a DNase for apurinic/apyrimidinic sites. The induction was blocked by chloramphenicol. Increases of 3-fold or more were also seen with plumbagin, menadione, and phenazine methosulfate. H/sub 2/O/sub 2/ produced no more than a 2-fold increase in endonuclease IV activity. The following agents had no significant effect: streptonigrin, nitrofurantoin, tert-butyl hydroperoxide, ..gamma.. rays, 260-nm UV radiation, methyl methanesulfonate, mitomycin C, and ascorbate. Paraquat, plumbagin, menadione, and phenazine methosulfate are known to generate superoxide radical anions via redox cycling in vivo. A mutant lacking superoxide dismutase was unusually sensitive to induction by paraquat. In addition, endonuclease IV could be induced by merely growing the mutant in pure O/sub 2/. The levels of endonuclease IV in uninduced or paraquat-treated cells were unaffected by mutations of oxyR, a H/sub 2/O/sub 2/-inducible gene that governs an oxidative-stress regulon. The results indicate that endonuclease IV is an inducible DNA-repair enzyme and that its induction can be mediated via the production of superoxide radicals.

  7. Endonuclease IV of Escherichia coli is induced by paraquat

    International Nuclear Information System (INIS)

    Chan, E.; Weiss, B.

    1987-01-01

    The addition of paraquat (methyl viologen) to a growing culture of Escherichia coli K-12 led within 1 hr to a 10- to 20-fold increase in the level of endonuclease IV, a DNase for apurinic/apyrimidinic sites. The induction was blocked by chloramphenicol. Increases of 3-fold or more were also seen with plumbagin, menadione, and phenazine methosulfate. H 2 O 2 produced no more than a 2-fold increase in endonuclease IV activity. The following agents had no significant effect: streptonigrin, nitrofurantoin, tert-butyl hydroperoxide, γ rays, 260-nm UV radiation, methyl methanesulfonate, mitomycin C, and ascorbate. Paraquat, plumbagin, menadione, and phenazine methosulfate are known to generate superoxide radical anions via redox cycling in vivo. A mutant lacking superoxide dismutase was unusually sensitive to induction by paraquat. In addition, endonuclease IV could be induced by merely growing the mutant in pure O 2 . The levels of endonuclease IV in uninduced or paraquat-treated cells were unaffected by mutations of oxyR, a H 2 O 2 -inducible gene that governs an oxidative-stress regulon. The results indicate that endonuclease IV is an inducible DNA-repair enzyme and that its induction can be mediated via the production of superoxide radicals

  8. Exchange stiffness of Ca-doped YIG

    Science.gov (United States)

    Avgin, I.; Huber, D. L.

    1994-05-01

    An effective medium theory for the zero-temperature exchange stiffness of uncompensated Ca-doped YIG is presented. The theory is based on the assumption that the effect of the Ca impurities is to produce strong, random ferromagnetic interactions between spins on the a and d sublattices. In the simplest version of the theory, a fraction, x, of the ad exchange integrals are large and positive, x being related to the Ca concentration. The stiffness is calculated as function of x for arbitrary perturbed ad exchange integral, Jxad. For Jxad≳(1/5)‖8Jaa+3Jdd‖, with Jaa and Jdd denoting the aa and dd exchange integrals, respectively, there is a critical concentration, Xc, such that when x≳Xc, the stiffness is complex. It is suggested that Xc delineates the region where there are significant departures from colinearity in the ground state of the Fe spins. Extension of the theory to a model where the Ca doping is assumed to generate Fe4+ ions on the tetrahedral sites is discussed. Possible experimental tests of the theory are mentioned.

  9. Efficient coherent driving of NV centers in a YIG-nanodiamond hybrid platform

    Science.gov (United States)

    Andrich, Paolo; de Las Casas, Charles F.; Liu, Xiaoying; Bretscher, Hope L.; Nealey, Paul F.; Awschalom, David D.; Heremans, F. Joseph

    The nitrogen-vacancy (NV) center in diamond is an ideal candidate for room temperature quantum computing and sensing applications. These schemes rely on magnetic dipolar interactions between the NV centers and other paramagnetic centers, imposing a stringent limit on the spin-to-spin separation. For instance, creating multi-qubit entanglement requires two NV centers to be within a few nanometers of each other, limiting the possibility for individual optical and microwave (MW) control. Moreover, to sense spins external to the diamond lattice the NV centers need to be within few nanometers from the surface, where their coherence properties are strongly reduced. In this work, we address these limitations using a hybrid YIG-nanodiamond platform where propagating spin-waves (SWs) are used to mediate the interaction between a MW source and a NV center ensemble, thereby relaxing the requirements imposed by dipolar interactions. In particular, we show that SWs can be used to amplify a MW signal detected by the NV centers by more than two orders of magnitude, allowing us to obtain ultra-low energy SW-driven coherent control of the NV centers. These results demonstrate the potentials of YIG-ND hybrid systems for the realization of enhanced quantum sensing and scalable computing devices. This work is supported by the ARO MURI program and the AFOSR.

  10. Endonucleases induced TRAIL-insensitive apoptosis in ovarian carcinoma cells

    Energy Technology Data Exchange (ETDEWEB)

    Geel, Tessa M. [Department of Pathology and Medical Biology, Groningen University Institute for Drug Exploration (GUIDE), University Medical Center Groningen (UMCG), Hanzeplein 1, 9713 GZ, Groningen (Netherlands); Meiss, Gregor [Institute of Biochemistry, Justus-Liebig-University Giessen, D-35392 Giessen (Germany); Gun, Bernardina T. van der; Kroesen, Bart Jan; Leij, Lou F. de [Department of Pathology and Medical Biology, Groningen University Institute for Drug Exploration (GUIDE), University Medical Center Groningen (UMCG), Hanzeplein 1, 9713 GZ, Groningen (Netherlands); Zaremba, Mindaugas; Silanskas, Arunas [Institute of Biotechnology, Vilnius LT-02241 (Lithuania); Kokkinidis, Michael [IMBB/FORTH and University of Crete/Department of Biology, GR-71409 Heraklion/Crete (Greece); Pingoud, Alfred [Institute of Biochemistry, Justus-Liebig-University Giessen, D-35392 Giessen (Germany); Ruiters, Marcel H. [Department of Pathology and Medical Biology, Groningen University Institute for Drug Exploration (GUIDE), University Medical Center Groningen (UMCG), Hanzeplein 1, 9713 GZ, Groningen (Netherlands); Synvolux therapeutics, Groningen (Netherlands); McLaughlin, Pamela M. [Department of Pathology and Medical Biology, Groningen University Institute for Drug Exploration (GUIDE), University Medical Center Groningen (UMCG), Hanzeplein 1, 9713 GZ, Groningen (Netherlands); Rots, Marianne G., E-mail: m.g.rots@med.umcg.nl [Department of Pathology and Medical Biology, Groningen University Institute for Drug Exploration (GUIDE), University Medical Center Groningen (UMCG), Hanzeplein 1, 9713 GZ, Groningen (Netherlands)

    2009-09-10

    TRAIL induced apoptosis of tumor cells is currently entering phase II clinical settings, despite the fact that not all tumor types are sensitive to TRAIL. TRAIL resistance in ovarian carcinomas can be caused by a blockade upstream of the caspase 3 signaling cascade. We explored the ability of restriction endonucleases to directly digest DNA in vivo, thereby circumventing the caspase cascade. For this purpose, we delivered enzymatically active endonucleases via the cationic amphiphilic lipid SAINT-18{sup Registered-Sign }:DOPE to both TRAIL-sensitive and insensitive ovarian carcinoma cells (OVCAR and SKOV-3, respectively). Functional nuclear localization after delivery of various endonucleases (BfiI, PvuII and NucA) was indicated by confocal microscopy and genomic cleavage analysis. For PvuII, analysis of mitochondrial damage demonstrated extensive apoptosis both in SKOV-3 and OVCAR. This study clearly demonstrates that cellular delivery of restriction endonucleases holds promise to serve as a novel therapeutic tool for the treatment of resistant ovarian carcinomas.

  11. Identification of a mismatch-specific endonuclease in hyperthermophilic Archaea.

    Science.gov (United States)

    Ishino, Sonoko; Nishi, Yuki; Oda, Soichiro; Uemori, Takashi; Sagara, Takehiro; Takatsu, Nariaki; Yamagami, Takeshi; Shirai, Tsuyoshi; Ishino, Yoshizumi

    2016-04-20

    The common mismatch repair system processed by MutS and MutL and their homologs was identified in Bacteria and Eukarya. However, no evidence of a functional MutS/L homolog has been reported for archaeal organisms, and it is not known whether the mismatch repair system is conserved in Archaea. Here, we describe an endonuclease that cleaves double-stranded DNA containing a mismatched base pair, from the hyperthermophilic archaeon Pyrococcus furiosus The corresponding gene revealed that the activity originates from PF0012, and we named this enzyme Endonuclease MS (EndoMS) as the mismatch-specific Endonuclease. The sequence similarity suggested that EndoMS is the ortholog of NucS isolated from Pyrococcus abyssi, published previously. Biochemical characterizations of the EndoMS homolog from Thermococcus kodakarensis clearly showed that EndoMS specifically cleaves both strands of double-stranded DNA into 5'-protruding forms, with the mismatched base pair in the central position. EndoMS cleaves G/T, G/G, T/T, T/C and A/G mismatches, with a more preference for G/T, G/G and T/T, but has very little or no effect on C/C, A/C and A/A mismatches. The discovery of this endonuclease suggests the existence of a novel mismatch repair process, initiated by the double-strand break generated by the EndoMS endonuclease, in Archaea and some Bacteria. © The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.

  12. Endonuclease α from Saccharomyces cerevisiae shows increased activity on ultraviolet irradiated native DNA

    International Nuclear Information System (INIS)

    Bryant, D.W.; Haynes, R.H.

    1978-01-01

    Endonuclease α isolated from the nucleus of the yeast Saccharomyces cerevisiae is a DNA endonuclease which has been shown to act preferentially on denatured T7 DNA. The purified enzyme is more active with UV-irradiated native T7 DNA than with unirradiated substrate. The relation between damage, measured by pyrimidine dimer concentration, and excess endonuclease activity is most readily explained by local denaturation caused by the presence of pyrimidine dimers. When three radiation sensitive mutants of yeast were tested for the level of endonuclease α present, none were found lacking the enzyme. However, nuclei of strain rad 1-1, a mutant that may be defective in heteroduplex repair as well as excision repair, were found to contain reduced levels of the endonuclease. (orig./AJ) [de

  13. The Effect of Ca–Zr mole ratio on preparation, structural and magnetic properties of Ca–Zr doped Bi-YIG

    International Nuclear Information System (INIS)

    Hasanpour, A.; Niyaifar, M.; Faridniya, N.; Amighian, J.

    2013-01-01

    Graphical abstract: - Highlights: • Ca–Zr substituted Bi-YIG has been prepared via mechanochemical processing and heat treatment. • Structural and magnetic properties were investigated. • Magnetic measurements showed the Ferrimagnetic behavior for pure garnet structure samples. • The spherical morphology of the nanoparticles was found from the SEM micrograph. - Abstract: Ca–Zr substituted (Bi-YIG) nanopowders with a nominal composition of BiY 2−x Ca x Zr y Fe 5−y O 12 (x = y and x varied from 0.00 to 1.25 by the step of 0.25) were prepared by mechanochemical processing (MCP) and subsequent heat treatments. The effect of dopant mol ratios, on garnet phase formation were investigated by X-ray diffraction (XRD) and Fourier transform infrared (FT-IR) spectroscopy. The lattice constant of the samples increased by increasing Zr 4+ content (for x ≤ 1). Mean crystallite size of the single-phase powders, which was evaluated by Scherrer's formula, was about 35 nm. The experimental results show that the Ca–Zr substitution Bi-YIG lowers the calcining and sintering temperatures for x < 1. The results show that the single-phase nanopowders can be obtained at temperatures below 850 °C. The measurements of vibrating sample magnetometer (VSM) show that the saturation magnetization of the samples increases as x increase from 0.00 to 0.25 and then decreases by increasing x to the values greater than 0.25

  14. HIGH TEMPERATURE RESONANCE LOSSES IN SILICON-DOPED YTTRIUM-IRON GARNET (YIG)

    DEFF Research Database (Denmark)

    Epstein, D. J.; Tocci, L.

    1967-01-01

    The ferrimagnetic resonance linewidth of silicon-doped YIG, measured as a function of temperature at 13.4 kMHz, is found to show a pronounced peak at 105°C. The anisotropic behavior of this peak is in good agreement with the four-level valence-exchange model proposed by Clogston. The model yields...... for the electron ordering energy a value 5 × 10-4 eV which agrees closely with the energy deduced from magnetic anneal studies. The activation energy for electron transfer (0.25 eV) is virtually identical with values reported in investigations of electrical conductivity and acoustic loss. ©1967 The American...

  15. Binding of T4 endonuclease V to deoxyribonucleic acid irradiated with ultraviolet light

    International Nuclear Information System (INIS)

    Seawell, P.C.; Simon, T.J.; Ganesan, A.K.

    1980-01-01

    Endonuclease V of bacteriophage T4 binds to uv-irradiated deoxyribonucleic acid (DNA) but not to unirradiated DNA. We have developed an assay to detect this binding, based on the retention of enzyme - DNA complexes on nitrocellulose filters. The amount of complex retained, ascertained by using radioactive DNA, is a measure of T4 endonuclease V activity. From our data we conclude that (1) T4 endonuclease V binds to uv-irradiated DNA but not to DNA that has been previously incised by the endonuclease, (2) equilibrium between the free and complexed form of the enzyme is attained under our reaction conditions, (3) dissociation of enzyme - DNA complexes is retarded by sodium cyanide, and (4) retention of enzyme - DNA complexes on nitrocellulose filters is enhanced by high concentrations of saline-citrate

  16. Combined Brillouin light scattering and microwave absorption study of magnon-photon coupling in a split-ring resonator/YIG film system

    Energy Technology Data Exchange (ETDEWEB)

    Klingler, S., E-mail: stefan.klingler@wmi.badw.de; Maier-Flaig, H.; Weiler, M. [Walther-Meißner-Institut, Bayerische Akademie der Wissenschaften, Walther-Meißner-Straße 8, 85748 Garching (Germany); Physik-Department, Technische Universität München, 85748 Garching (Germany); Gross, R.; Huebl, H.; Goennenwein, S. T. B. [Walther-Meißner-Institut, Bayerische Akademie der Wissenschaften, Walther-Meißner-Straße 8, 85748 Garching (Germany); Physik-Department, Technische Universität München, 85748 Garching (Germany); Nanosystems Initiative Munich (NIM), 80799 Munich (Germany); Hu, C.-M. [Department of Physics and Astronomy, University of Manitoba, Winnipeg, Manitoba R3T2N2 (Canada)

    2016-08-15

    Microfocused Brillouin light scattering (BLS) and microwave absorption (MA) are used to study magnon-photon coupling in a system consisting of a split-ring microwave resonator and an yttrium iron garnet (YIG) film. The split-ring resonator is defined by optical lithography and loaded with a 1 μm-thick YIG film grown by liquid phase epitaxy. BLS and MA spectra of the hybrid system are simultaneously recorded as a function of the applied magnetic field magnitude and microwave excitation frequency. Strong coupling of the magnon and microwave resonator modes is found with a coupling strength of g{sub eff} /2π = 63 MHz. The combined BLS and MA data allow us to study the continuous transition of the hybridized modes from a purely magnonic to a purely photonic mode by varying the applied magnetic field and microwave frequency. Furthermore, the BLS data represent an up-conversion of the microwave frequency coupling to optical frequencies.

  17. Simple and sensitive fluorescence assay of restriction endonuclease on graphene oxide

    International Nuclear Information System (INIS)

    Gang, Jong Back

    2015-01-01

    Restriction endonucleases hydrolyze internal phosphodiester bonds at specific sites in a DNA sequence. These enzymes are essential in a variety of fields, such as biotechnology and clinical diagnostics. It is of great importance and necessity for the scientific and biomedical use of enzymes to measure endonuclease activity. In this study, graphene oxide (GO) has been used as a platform to measure enzyme activity with high sensitivity. To increase the detection sensitivity of Hinf I, the endonuclease-digested reaction was treated with exonuclease III (Exo III) and a fluorescence assay was conducted to measure the emission. Results showed that Exo III treatment enhanced 2.7-fold signal-to-background ratio for the detection of Hinf I compared with that done without Exo III in the presence of GO

  18. Properties of an endonuclease activity in Micrococcus luteus acting on γ-irradiated DNA and on apurinic DNA

    International Nuclear Information System (INIS)

    Schaefer, G.; Haas, P.; Coquerelle, Th.; Hagen, U.

    1980-01-01

    A protein fraction from Micrococcus luteus with endonuclease activity against γ-irradiated DNA was isolated and characterized. An additional activity on apurinic sites could not be separated, either by sucrose gradient sedimentation or by gel filtration through Sephadex G 100. From gel filtration, a molecular weight of about 25 000 was calculated for both endonuclease activities. The endonuclease activity against γ-irradiated DNA was stimulated five-fold with 5 mM Mg ++ , whereas that against apurinic sites was less dependent on the Mg ++ concentration. 100 mM KCl inhibited the γ-ray endonuclease, but not the apurinic endonuclease activity. In γ-irradiated DNA the protein recognized 1.65 endonuclease sensitive sites per radiation-induced single-strand break, among which are 0.45 alkali labile lesions in the nucleotide strand. The was evaluated resulting in a Ksub(m)-value of 73 nM. (author)

  19. Substrate specificity of Micrococcus luteus uv endonuclease and its overlap with DNA photolyase activity

    International Nuclear Information System (INIS)

    Patrick, M.H.

    1975-01-01

    The action of an endonuclease from Micrococcus luteus that operates on uv damage in DNA overlaps with that of DNA photolyase from yeast: homo- and heterocyclobutane dipyrimidines in DNA are substrates for both enzymes, but pyrimidine adducts or the spore photoproduct in DNA are not. As expected from this overlap, the action of the two enzymes is mutually interfering: single-strand nicks introduced by the endonuclease effectively preclude photoreactivation; conversely, formation of a photolyase-cyclobutane dipyrimidine complex can prevent nicking by the endonuclease

  20. Properties of an endonuclease activity in Micrococcus luteus acting on. gamma. -irradiated DNA and on apurinic DNA

    Energy Technology Data Exchange (ETDEWEB)

    Schaefer, G; Haas, P; Coquerelle, Th; Hagen, U [Kernforschungszentrum Karlsruhe G.m.b.H. (Germany, F.R.). Inst. fuer Genetik und fuer Toxikologie von Spaltstoffen

    1980-01-01

    A protein fraction from Micrococcus luteus with endonuclease activity against ..gamma..-irradiated DNA was isolated and characterized. An additional activity on apurinic sites could not be separated, either by sucrose gradient sedimentation or by gel filtration through Sephadex G 100. From gel filtration, a molecular weight of about 25 000 was calculated for both endonuclease activities. The endonuclease activity against ..gamma..-irradiated DNA was stimulated five-fold with 5 mM Mg/sup + +/, whereas that against apurinic sites was less dependent on the Mg/sup + +/ concentration. 100 mM KCl inhibited the ..gamma..-ray endonuclease, but not the apurinic endonuclease activity. In ..gamma..-irradiated DNA the protein recognized 1.65 endonuclease sensitive sites per radiation-induced single-strand break, among which are 0.45 alkali labile lesions in the nucleotide strand. The was evaluated resulting in a Ksub(m)-value of 73 nM.

  1. Effect of magnetization boundary condition on cavity magnon polariton of YIG thin film

    Science.gov (United States)

    Jiang, H. H.; Xiao, Y.; Hu, C. M.; Guo, H.; Xia, K.

    2018-06-01

    Motivated by recent studies of cavity magnon polariton (CMP), we extended a previous theoretical work to generalize microwave transmission calculation with various magnetization boundary condition of YIG thin film embedded in cavity. It is found that numerical implementation given in this paper can be easily applied to other magnetization boundary condition and extended to magnetic multilayers. Numerical results show that ferromagnetic resonance mode of microwave transmission spectrum, which is absent in previous calculation, can be recovered by altering the pinning condition of surface spins. The demonstrated reliability of our theory opens attractive perspectives for studying CMP of thin film with complicated surface magnetization distribution and magnetic multilayers.

  2. Spin Seebeck effect in nanometer-thick YIG micro-fabricated strips

    Directory of Open Access Journals (Sweden)

    Martin Collet

    2017-05-01

    Full Text Available We have investigated the spin Seebeck effect (SSE generated by current induced-heating in ultra-thin yttrium iron garnet film (20 nm covered by an 8 nm thick Pt layer. By passing current in the Pt layer, an out-of-plane temperature gradient is established that, in turn, generates an out-of-equilibrium magnons population. The resulting pure spin current is detected using the inverse spin Hall effect (ISHE measured in the Pt electrode. A lock-in detection scheme is used to separate the SSE signal from other magneto-galvanic effect. Indeed, the SSE signal is obtained as the second harmonic voltage response, while spin Hall magnetoresistance (SMR is measured as the first harmonic response to the ac excitation current. Interestingly, the amplitude of the SSE in such thin YIG film is comparable to what has been reported for much thicker films.

  3. Processive nicking activity of T4 endonuclease V on UV-irradiated chromatin

    International Nuclear Information System (INIS)

    Gruskin, E.A.; Lloyd, R.S.

    1986-01-01

    T4 endonuclease V initiates the excision repair of pyrimidine dimers in UV-irradiated T4 infected E. coli cells. The pyrimidine dimer specific nicking activity of T4 endonuclease V functions by a processive scanning on UV-irradiated DNA. Previously it has been demonstrated that introduction of endonuclease V into repair-deficient human cells causes a restoration of UV survival in these cells. This demonstrates that endonuclease V is competent to incise mammalian DNA at the site of pyrimidine dimers. In order to assess the ability of endonuclease V to act processively on DNA associated as chromatin, minichromosomes were prepared for use as a substrate. Form I DNA was reconstituted with H3, H4 +/- H1 histones by sequential dialysis steps from 2.0 M NaCl to 50 mM NaCl. Time course reactions were performed with minichromosomes containing 10 and 25 dimers per molecule. In each case the rate of disappearance of form I DNA which was associated as chromatin was decreased relative to that of naked form I DNA. Concurrent with that observation, the rate and extent of appearance of form III DNA was increased with the DNA in minichromosomes relative to naked DNA. This is diagnostic of an enhancement of processivity. The inclusion of H1 in the minichromosomes resulted in a slight additional increase in processivity relative to minichromosomes consisting only of H3 and H4

  4. The metabolic enhancer piracetam attenuates mitochondrion-specific endonuclease G translocation and oxidative DNA fragmentation.

    Science.gov (United States)

    Gupta, Sonam; Verma, Dinesh Kumar; Biswas, Joyshree; Rama Raju, K Siva; Joshi, Neeraj; Wahajuddin; Singh, Sarika

    2014-08-01

    This study was performed to investigate the involvement of mitochondrion-specific endonuclease G in piracetam (P)-induced protective mechanisms. Studies have shown the antiapoptotic effects of piracetam but the mechanism of action of piracetam is still an enigma. To assess the involvement of endonuclease G in piracetam-induced protective effects, astrocyte glial cells were treated with lipopolysaccharide (LPS) and piracetam. LPS treatment caused significantly decreased viability, mitochondrial activity, oxidative stress, chromatin condensation, and DNA fragmentation, which were attenuated by piracetam cotreatment. Cotreatment of astrocytes with piracetam showed its significantly time-dependent absorption as observed with high-performance liquid chromatography. Astrocytes treated with piracetam alone showed enhanced mitochondrial membrane potential (MMP) in comparison to control astrocytes. However, in LPS-treated cells no significant alteration in MMP was observed in comparison to control cells. Protein and mRNA levels of the terminal executor of the caspase-mediated pathway, caspase-3, were not altered significantly in LPS or LPS + piracetam-treated astrocytes, whereas endonuclease G was significantly translocated to the nucleus in LPS-treated astrocytes. Piracetam cotreatment attenuated the LPS-induced endonuclease G translocation. In conclusion this study indicates that LPS treatment of astrocytes caused decreased viability, oxidative stress, mitochondrial dysfunction, chromatin condensation, DNA damage, and translocation of endonuclease G to the nucleus, which was inhibited by piracetam cotreatment, confirming that the mitochondrion-specific endonuclease G is one of the factors involved in piracetam-induced protective mechanisms. Copyright © 2014 Elsevier Inc. All rights reserved.

  5. A functional endonuclease Q exists in the bacterial domain: identification and characterization of endonuclease Q from Bacillus pumilus.

    Science.gov (United States)

    Shiraishi, Miyako; Ishino, Sonoko; Cann, Isaac; Ishino, Yoshizumi

    2017-05-01

    DNA base deamination occurs spontaneously under physiological conditions and is promoted by high temperature. Therefore, hyperthermophiles are expected to have efficient repair systems of the deaminated bases in their genomes. Endonuclease Q (EndoQ) was originally identified from the hyperthermophlic archaeon, Pyrococcus furiosus, as a hypoxanthine-specific endonuclease recently. Further biochemical analyses revealed that EndoQ also recognizes uracil, xanthine, and the AP site in DNA, and is probably involved in a specific repair process for damaged bases. Initial phylogenetic analysis showed that an EndoQ homolog is found only in the Thermococcales and some of the methanogens in Archaea, and is not present in most members of the domains Bacteria and Eukarya. A better understanding of the distribution of the EndoQ-mediated repair system is, therefore, of evolutionary interest. We showed here that an EndoQ-like polypeptide from Bacillus pumilus, belonging to the bacterial domain, is functional and has similar properties with the archaeal EndoQs.

  6. DNA Nucleotide Sequence Restricted by the RI Endonuclease

    Science.gov (United States)

    Hedgpeth, Joe; Goodman, Howard M.; Boyer, Herbert W.

    1972-01-01

    The sequence of DNA base pairs adjacent to the phosphodiester bonds cleaved by the RI restriction endonuclease in unmodified DNA from coliphage λ has been determined. The 5′-terminal nucleotide labeled with 32P and oligonucleotides up to the heptamer were analyzed from a pancreatic DNase digest. The following sequence of nucleotides adjacent to the RI break made in λ DNA was deduced from these data and from the 3′-dinucleotide sequence and nearest-neighbor analysis obtained from repair synthesis with the DNA polymerase of Rous sarcoma virus [Formula: see text] The RI endonuclease cleavage of the phosphodiester bonds (indicated by arrows) generates 5′-phosphoryls and short cohesive termini of four nucleotides, pApApTpT. The most striking feature of the sequence is its symmetry. PMID:4343974

  7. Efficient fdCas9 Synthetic Endonuclease with Improved Specificity for Precise Genome Engineering

    KAUST Repository

    Aouida, Mustapha

    2015-07-30

    The Cas9 endonuclease is used for genome editing applications in diverse eukaryotic species. A high frequency of off-target activity has been reported in many cell types, limiting its applications to genome engineering, especially in genomic medicine. Here, we generated a synthetic chimeric protein between the catalytic domain of the FokI endonuclease and the catalytically inactive Cas9 protein (fdCas9). A pair of guide RNAs (gRNAs) that bind to sense and antisense strands with a defined spacer sequence range can be used to form a catalytically active dimeric fdCas9 protein and generate double-strand breaks (DSBs) within the spacer sequence. Our data demonstrate an improved catalytic activity of the fdCas9 endonuclease, with a spacer range of 15–39 nucleotides, on surrogate reporters and genomic targets. Furthermore, we observed no detectable fdCas9 activity at known Cas9 off-target sites. Taken together, our data suggest that the fdCas9 endonuclease variant is a superior platform for genome editing applications in eukaryotic systems including mammalian cells.

  8. Efficient fdCas9 Synthetic Endonuclease with Improved Specificity for Precise Genome Engineering

    KAUST Repository

    Aouida, Mustapha; Eid, Ayman; Ali, Zahir; Cradick, Thomas; Lee, Ciaran; Deshmukh, Harshavardhan; Atef, Ahmed; Abu Samra, Dina Bashir Kamil; Gadhoum, Samah Zeineb; Merzaban, Jasmeen; Bao, Gang; Mahfouz, Magdy M.

    2015-01-01

    The Cas9 endonuclease is used for genome editing applications in diverse eukaryotic species. A high frequency of off-target activity has been reported in many cell types, limiting its applications to genome engineering, especially in genomic medicine. Here, we generated a synthetic chimeric protein between the catalytic domain of the FokI endonuclease and the catalytically inactive Cas9 protein (fdCas9). A pair of guide RNAs (gRNAs) that bind to sense and antisense strands with a defined spacer sequence range can be used to form a catalytically active dimeric fdCas9 protein and generate double-strand breaks (DSBs) within the spacer sequence. Our data demonstrate an improved catalytic activity of the fdCas9 endonuclease, with a spacer range of 15–39 nucleotides, on surrogate reporters and genomic targets. Furthermore, we observed no detectable fdCas9 activity at known Cas9 off-target sites. Taken together, our data suggest that the fdCas9 endonuclease variant is a superior platform for genome editing applications in eukaryotic systems including mammalian cells.

  9. A new restriction endonuclease from Citrobacter freundii

    OpenAIRE

    Janulaitis, A.A.; Stakenas, P.S.; Lebedenko, E.N.; Berlin, Yu.A.

    1982-01-01

    CfrI, a new restriction endonuclease of unique substrate specificity, has been isolated from a Citrobacter freundii strain. The enzyme recognizes a degenerated sequence PyGGCCPu in double-strand DNA and cleaves it between Py and G residues to yield 5′ -protruding tetranucleotide ends GGCC.

  10. A new restriction endonuclease from Citrobacter freundii

    Science.gov (United States)

    Janulaitis, A.A.; Stakenas, P.S.; Lebedenko, E.N.; Berlin, Yu.A.

    1982-01-01

    CfrI, a new restriction endonuclease of unique substrate specificity, has been isolated from a Citrobacter freundii strain. The enzyme recognizes a degenerated sequence PyGGCCPu in double-strand DNA and cleaves it between Py and G residues to yield 5′ -protruding tetranucleotide ends GGCC. Images PMID:6294607

  11. Comparison of genomes of malignant catarrhal fever-associated herpesviruses by restriction endonuclease analysis.

    Science.gov (United States)

    Shih, L M; Zee, Y C; Castro, A E

    1989-01-01

    The restriction endonuclease DNA cleavage patterns of eight isolates of malignant catarrhal fever-associated herpesviruses were examined using the restriction endonucleases HindIII and EcoRI. The eight viruses could be assigned to two distinct groups. Virus isolates from a blue wildebeest, a sika deer and an ibex had restriction endonuclease DNA cleavage patterns that were in general similar to each other. The restriction pattern of these three viruses was distinct from the other five. Of these five, four were isolated from a greater kudu, a white tailed wildebeest, a white bearded wildebeest, and a cape hartebeest. The fifth isolate C500, was isolated from a domestic cow with malignant catarrhal fever. These five viruses had similar DNA cleavage patterns.

  12. Excision repair of gamma-ray-induced alkali-stable DNA lesions with the help of γ-endonuclease from Micrococcus luteus

    International Nuclear Information System (INIS)

    Tomilin, N.V.; Barenfeld, L.S.

    1979-01-01

    γ-endonuclease Y, an enzyme that hydrolyses phosphodiester bonds at alkali-stable lesions in γ-irradiated (N 2 , tris buffer) DNA, has been partially purified from Micrococcus luteus. The enzyme has a molecular weight of about 19 000, induces single-strand breaks with 3'OH-5'PO 4 termini and contains endonuclease activity towards DNA treated with 7-bromomethylbenz(a)anthracene. γ-endonuclease Y induces breaks in OsO 4 -treated poly(dA-dT) and apparently is specific towards γ-ray-induced base lesions of the t' type. The complete excision repair of γ-endonuclease Y substrate sites has been performed in vitro by γ-endonuclease Y, DNA polymerase and ligase. (author)

  13. Excision repair of gamma-ray-induced alkali-stable DNA lesions with the help of. gamma. -endonuclease from Micrococcus luteus

    Energy Technology Data Exchange (ETDEWEB)

    Tomilin, N V; Barenfeld, L S [AN SSSR, Leningrad. Inst. Tsitologii

    1979-03-01

    ..gamma..-endonuclease Y, an enzyme that hydrolyses phosphodiester bonds at alkali-stable lesions in ..gamma..-irradiated (N/sub 2/, tris buffer) DNA, has been partially purified from Micrococcus luteus. The enzyme has a molecular weight of about 19 000, induces single-strand breaks with 3'OH-5'PO/sub 4/ termini and contains endonuclease activity towards DNA treated with 7-bromomethylbenz(a)anthracene. ..gamma..-endonuclease Y induces breaks in OsO/sub 4/-treated poly(dA-dT) and apparently is specific towards ..gamma..-ray-induced base lesions of the t' type. The complete excision repair of ..gamma..-endonuclease Y substrate sites has been performed in vitro by ..gamma..-endonuclease Y, DNA polymerase and ligase.

  14. DENV gene of bacteriophage T4 codes for both pyrimidine dimer-DNA glycosylase and apyrimidinic endonuclease activities

    International Nuclear Information System (INIS)

    McMillan, S.; Edenberg, H.J.; Radany, E.H.; Friedberg, R.C.; Friedberg, E.C.

    1981-01-01

    Recent studies have shown that purified preparations of phage T4 UV DNA-incising activity (T4 UV endonuclease or endonuclease V of phase T4) contain a pyrimidine dimer-DNA glycosylase activity that catalyzes hydrolysis of the 5' glycosyl bond of dimerized pyrimidines in UV-irradiated DNA. Such enzyme preparations have also been shown to catalyze the hydrolysis of phosphodiester bonds in UV-irradiated DNA at a neutral pH, presumably reflecting the action of an apurinic/apyrimidinic endonuclease at the apyrimidinic sites created by the pyrimidine dimer-DNA glycosylase. In this study we found that preparations of T4 UV DNA-incising activity contained apurinic/apyrimidinic endonuclease activity that nicked depurinated form I simian virus 40 DNA. Apurinic/apyrimidinic endonuclease activity was also found in extracts of Escherichia coli infected with T4 denV + phage. Extracts of cells infected with T4 denV mutants contained significantly lower levels of apurinic/apyrimidinic endonuclease activity; these levels were no greater than the levels present in extracts of uninfected cells. Furthermore, the addition of DNA containing UV-irradiated DNA and T4 enzyme resulted in competition for pyrimidine dimer-DNA glycosylase activity against the UV-irradiated DNA. On the basis of these results, we concluded that apurinic/apyrimidinic endonuclease activity is encoded by the denV gene of phage T4, the same gene that codes for pyrimidine dimer-DNA glycosylase activity

  15. Structural aspects of catalytic mechanisms of endonucleases and their binding to nucleic acids

    Energy Technology Data Exchange (ETDEWEB)

    Zhukhlistova, N. E.; Balaev, V. V.; Lyashenko, A. V.; Lashkov, A. A., E-mail: alashkov83@gmail.com [Russian Academy of Sciences, Shubnikov Institute of Crystallography (Russian Federation)

    2012-05-15

    Endonucleases (EC 3.1) are enzymes of the hydrolase class that catalyze the hydrolytic cleavage of deoxyribonucleic and ribonucleic acids at any region of the polynucleotide chain. Endonucleases are widely used both in biotechnological processes and in veterinary medicine as antiviral agents. Medical applications of endonucleases in human cancer therapy hold promise. The results of X-ray diffraction studies of the spatial organization of endonucleases and their complexes and the mechanism of their action are analyzed and generalized. An analysis of the structural studies of this class of enzymes showed that the specific binding of enzymes to nucleic acids is characterized by interactions with nitrogen bases and the nucleotide backbone, whereas the nonspecific binding of enzymes is generally characterized by interactions only with the nucleic-acid backbone. It should be taken into account that the specificity can be modulated by metal ions and certain low-molecular-weight organic compounds. To test the hypotheses about specific and nonspecific nucleic-acid-binding proteins, it is necessary to perform additional studies of atomic-resolution three-dimensional structures of enzyme-nucleic-acid complexes by methods of structural biology.

  16. Characterization of DNA substrate specificities of apurinic/apyrimidinic endonucleases from Mycobacterium tuberculosis.

    Science.gov (United States)

    Abeldenov, Sailau; Talhaoui, Ibtissam; Zharkov, Dmitry O; Ishchenko, Alexander A; Ramanculov, Erlan; Saparbaev, Murat; Khassenov, Bekbolat

    2015-09-01

    Apurinic/apyrimidinic (AP) endonucleases are key enzymes involved in the repair of abasic sites and DNA strand breaks. Pathogenic bacteria Mycobacterium tuberculosis contains two AP endonucleases: MtbXthA and MtbNfo members of the exonuclease III and endonuclease IV families, which are exemplified by Escherichia coli Xth and Nfo, respectively. It has been shown that both MtbXthA and MtbNfo contain AP endonuclease and 3'→5' exonuclease activities. However, it remains unclear whether these enzymes hold 3'-repair phosphodiesterase and nucleotide incision repair (NIR) activities. Here, we report that both mycobacterial enzymes have 3'-repair phosphodiesterase and 3'-phosphatase, and MtbNfo contains in addition a very weak NIR activity. Interestingly, depending on pH, both enzymes require different concentrations of divalent cations: 0.5mM MnCl2 at pH 7.6 and 10 mM at pH 6.5. MtbXthA requires a low ionic strength and 37 °C, while MtbNfo requires high ionic strength (200 mM KCl) and has a temperature optimum at 60 °C. Point mutation analysis showed that D180 and N182 in MtbXthA and H206 and E129 in MtbNfo are critical for enzymes activities. The steady-state kinetic parameters indicate that MtbXthA removes 3'-blocking sugar-phosphate and 3'-phosphate moieties at DNA strand breaks with an extremely high efficiency (kcat/KM=440 and 1280 μM(-1)∙min(-1), respectively), while MtbNfo exhibits much lower 3'-repair activities (kcat/KM=0.26 and 0.65 μM(-1)∙min(-1), respectively). Surprisingly, both MtbXthA and MtbNfo exhibited very weak AP site cleavage activities, with kinetic parameters 100- and 300-fold lower, respectively, as compared with the results reported previously. Expression of MtbXthA and MtbNfo reduced the sensitivity of AP endonuclease-deficient E. coli xth nfo strain to methylmethanesulfonate and H2O2 to various degrees. Taken together, these data establish the DNA substrate specificity of M. tuberculosis AP endonucleases and suggest their possible role

  17. A new endonuclease recognizing the deoxynucleotide sequence CCNNGG from the cyanobacterium Synechocystis 6701.

    Science.gov (United States)

    Calléja, F; Tandeau de Marsac, N; Coursin, T; van Ormondt, H; de Waard, A

    1985-09-25

    A new sequence-specific endonuclease from the cyanobacterium Synechocystis species PCC 6701 has been purified and characterized. This enzyme, SecI, is unique in recognizing the nucleotide sequence: 5' -CCNNGG-3' 3' -GGNNCC-5' and cleaves it at the position indicated by the symbol. Two other restriction endonucleases, SecII and SecIII, found in this organism are isoschizomers of MspI and MstII, respectively.

  18. PMS2 endonuclease activity has distinct biological functions and is essential for genome maintenance.

    Science.gov (United States)

    van Oers, Johanna M M; Roa, Sergio; Werling, Uwe; Liu, Yiyong; Genschel, Jochen; Hou, Harry; Sellers, Rani S; Modrich, Paul; Scharff, Matthew D; Edelmann, Winfried

    2010-07-27

    The DNA mismatch repair protein PMS2 was recently found to encode a novel endonuclease activity. To determine the biological functions of this activity in mammals, we generated endonuclease-deficient Pms2E702K knock-in mice. Pms2EK/EK mice displayed increased genomic mutation rates and a strong cancer predisposition. In addition, class switch recombination, but not somatic hypermutation, was impaired in Pms2EK/EK B cells, indicating a specific role in Ig diversity. In contrast to Pms2-/- mice, Pms2EK/EK male mice were fertile, indicating that this activity is dispensable in spermatogenesis. Therefore, the PMS2 endonuclease activity has distinct biological functions and is essential for genome maintenance and tumor suppression.

  19. Adenosine triphosphate stimulates Aquifex aeolicus MutL endonuclease activity.

    Directory of Open Access Journals (Sweden)

    Jerome Mauris

    2009-09-01

    Full Text Available Human PMS2 (hPMS2 homologues act to nick 5' and 3' to misincorporated nucleotides during mismatch repair in organisms that lack MutH. Mn(++ was previously found to stimulate the endonuclease activity of these homologues. ATP was required for the nicking activity of hPMS2 and yPMS1, but was reported to inhibit bacterial MutL proteins from Thermus thermophilus and Aquifex aeolicus that displayed homology to hPMS2. Mutational analysis has identified the DQHA(X(2E(X(4E motif present in the C-terminus of PMS2 homologues as important for endonuclease activity.We examined the effect ATP had on the Mn(++ induced nicking of supercoiled pBR322 by full-length and mutant A. aeolicus MutL (Aae MutL proteins. Assays were single time point, enzyme titration experiments or reaction time courses. The maximum velocity for MutL nicking was determined to be 1.6+/-0.08x10(-5 s(-1 and 4.2+/-0.3x10(-5 s(-1 in the absence and presence of ATP, respectively. AMPPNP stimulated the nicking activity to a similar extent as ATP. A truncated Aae MutL protein composed of only the C-terminal 123 amino acid residues was found to nick supercoiled DNA. Furthermore, mutations in the conserved C-terminal DQHA(X(2E(X(4E and CPHGRP motifs were shown to abolish Aae MutL endonuclease activity.ATP stimulated the Mn(++ induced endonuclease activity of Aae MutL. Experiments utilizing AMPPNP implied that the stimulation did not require ATP hydrolysis. A mutation in the DQHA(X(2E(X(4E motif of Aae MutL further supported the role of this region in endonclease activity. For the first time, to our knowledge, we demonstrate that changing the histidine residue in the conserved CPHGRP motif abolishes endonucleolytic activity of a hPMS2 homologue. Finally, the C-terminal 123 amino acid residues of Aae MutL were sufficient to display Mn(++ induced nicking activity.

  20. X-ray detected magnetic resonance of YIG thin films in the nonlinear regime of spin waves

    Energy Technology Data Exchange (ETDEWEB)

    Goulon, J., E-mail: goulon@esrf.f [European Synchrotron Radiation Facility (ESRF), B.P. 220, F-38043 Grenoble Cedex (France); Rogalev, A.; Wilhelm, F.; Goujon, G. [European Synchrotron Radiation Facility (ESRF), B.P. 220, F-38043 Grenoble Cedex (France); Brouder, Ch. [Institut de Mineralogie et de Physique des Milieux Condenses, UMR-CNRS 7590, Universite Paris VI-VII, 4 place Jussieu, F-75252 Paris Cedex 05 (France); Yaresko, A. [Max Planck Institute for Solid State Research, Heisenbergstrasse 1, 70569 Stuttgart (Germany); Ben Youssef, J.; Indenbom, M.V. [Laboratoire de Magnetisme de Bretagne, CNRS FRE 2697, UFR Sciences et Techniques, F-29328 Brest Cedex (France)

    2010-08-15

    We discuss the information content of element/edge resolved X-ray detected magnetic resonance (XDMR) experiments carried out on yttrium iron garnet (YIG) thin films. Starting with a phenomenological approach, it is shown that the photoionisation of deep atomic core levels by circularly polarized X-rays can be used to probe the precession dynamics of spin or orbital magnetization components in empty final states of proper symmetry. Crude estimates of the opening angle of the uniform precession mode were tentatively deduced from the ratio of the XDMR and XMCD absorption cross-sections either at the iron or yttrium absorbing sites. The implications of the most recent experimental results collected at the ESRF are analyzed, keeping in mind that: (i) the Fe K-edge XDMR signal is largely dominated by the precession of orbital magnetization components at the tetrahedral iron sites; (ii) the Y L-edges XDMR signal essentially describes the precession of induced spin magnetization involving the 4d states of yttrium. In the magnetostatic regime, we produce clear experimental evidence of collective excitations of orbital magnetization waves, especially under high pumping power. Several coupling mechanisms could explain our observations, starting with pseudo-dipolar interactions in ferromagnetic systems. In ferrimagnetic systems in which orbital degeneracy and orbital ordering make the excitation of orbitons possible, one may envisage additional modes of excitation or relaxation of orbital magnetization waves. This interpretation looks fully consistent with the results of band structure calculations carried out recently on YIG with fully relativistic LMTO-LSDA methods.

  1. Sequence specificity of DNA cleavage by Micrococcus luteus γ endonuclease

    International Nuclear Information System (INIS)

    Hentosh, P.; Henner, W.D.; Reynolds, R.J.

    1985-01-01

    DNA fragments of defined sequence have been used to determine the sites of cleavage by γ-endonuclease activity in extracts prepared from Micrococcus luteus. End-labeled DNA restriction fragments of pBR322 DNA that had been irradiated under nitrogen in the presence of potassium iodide or t-butanol were treated with M. luteus γ endonuclease and analyzed on irradiated DNA preferentially at the positions of cytosines and thymines. DNA cleavage occurred immediately to the 3' side of pyrimidines in irradiated DNA and resulted in fragments that terminate in a 5'-phosphoryl group. These studies indicate that both altered cytosines and thymines may be important DNA lesions requiring repair after exposure to γ radiation

  2. Involvement of the VDE homing endonuclease and rapamycin in regulation of the Saccharomyces cerevisiae GSH11 gene encoding the high affinity glutathione transporter.

    Science.gov (United States)

    Miyake, Tsuyoshi; Hiraishi, Hiroyuki; Sammoto, Hiroyuki; Ono, Bun-Ichiro

    2003-10-10

    The Saccharomyces cerevisiae gene HGT1/GSH11 encodes the high affinity glutathione transporter and is repressed by cysteine added to the culture medium. It has been found previously that a 5'-upstream cis-element, CCGCCACAC, is responsible for regulating GSH11 expression and that several proteins bind to this element (Miyake, T., Kanayama, M., Sammoto, H., and Ono, B. (2002) Mol. Genet. Genomics 266, 1004-1011). In this report we present evidence that the most prominent of these proteins is VDE, known previously as the homing endonuclease encoded by VMA1. We show also that GSH11 is not expressed in a VDE-deleted strain and that inability to express the GSH11 of this strain is overcome by introduction of the coding region of VDE or the entire VMA1 gene. It is also found that VDE does not cut DNA in the vicinity of the GSH11 cis-element. Rapamycin, an inhibitor of the target of rapamycin (TOR) signal-transduction system, is found to enhance expression of GSH11 in a VDE-dependent manner under conditions of sulfur starvation. These results indicate that GSH11 is regulated by a system sensitive to sulfur starvation (presumably via cysteine depletion) and a more general system involving the nutritional starvation signal mediated by the TOR system. Both systems need to be operational (inhibition of TOR and sulfur starvation) for full expression of GSH11.

  3. Purification, crystallization and preliminary crystallographic analysis of a thermostable endonuclease IV from Thermotoga maritima

    International Nuclear Information System (INIS)

    Hughes, Ronny C.; Tomanicek, Stephen J.; Ng, Joseph D.; Coates, Leighton

    2009-01-01

    The overexpression, purification and crystallization of endonuclease IV from T. maritima are reported. The crystals belonged to the hexagonal space group P6 1 and diffracted to 2.36 Å resolution. The DNA-repair enzyme endonuclease IV from the thermophilic bacterium Thermotoga maritima MSB8 (reference sequence NC-000853) has been expressed in Escherichia coli and crystallized for X-ray analysis. T. maritima endonuclease IV is a 287-amino-acid protein with 32% sequence identity to E. coli endonuclease IV. The protein was purified to homogeneity and was crystallized using the sitting-drop vapor-diffusion method. The protein crystallized in space group P6 1 , with one biological molecule in the asymmetric unit, corresponding to a Matthews coefficient of 2.39 Å 3 Da −1 and 47% solvent content. The unit-cell parameters of the crystals were a = b = 123.2, c = 35.6 Å. Microseeding and further optimization yielded crystals with an X-ray diffraction limit of 2.36 Å. A single 70° data set was collected and processed, resulting in an overall R merge and a completeness of 9.5% and 99.3%, respectively

  4. Expression analysis of a ''Cucurbita'' cDNA encoding endonuclease

    International Nuclear Information System (INIS)

    Szopa, J.

    1995-01-01

    The nuclear matrices of plant cell nuclei display intrinsic nuclease activity which consists in nicking supercoiled DNA. A cDNA encoding a 32 kDa endonuclease has been cloned and sequenced. The nucleotide and deduced amino-acid sequences show high homology to known 14-3-3-protein sequences from other sources. The amino-acid sequence shows agreement with consensus sequences for potential phosphorylation by protein kinase A and C and for calcium, lipid and membrane-binding sites. The nucleotide-binding site is also present within the conserved part of the sequence. By Northern blot analysis, the differential expression of the corresponding mRNA was detected; it was the strongest in sink tissues. The endonuclease activity found on DNA-polyacrylamide gel electrophoresis coincided with mRNA content and was the highest in tuber. (author). 22 refs, 6 figs

  5. Conserved structural chemistry for incision activity in structurally non-homologous apurinic/apyrimidinic endonuclease APE1 and endonuclease IV DNA repair enzymes.

    Energy Technology Data Exchange (ETDEWEB)

    Tsutakawa, Susan E.; Shin, David S.; Mol, Clifford D.; Izum, Tadahide; Arvai, Andrew S.; Mantha, Anil K.; Szczesny, Bartosz; Ivanov, Ivaylo N.; Hosfield, David J.; Maiti, Buddhadev; Pique, Mike E.; Frankel, Kenneth A.; Hitomi, Kenichi; Cunningham, Richard P.; Mitra, Sankar; Tainer, John A.

    2013-03-22

    Non-coding apurinic/apyrimidinic (AP) sites in DNA form spontaneously and as DNA base excision repair intermediates are the most common toxic and mutagenic in vivo DNA lesion. For repair, AP sites must be processed by 5' AP endonucleases in initial stages of base repair. Human APE1 and bacterial Nfo represent the two conserved 5' AP endonuclease families in the biosphere; they both recognize AP sites and incise the phosphodiester backbone 5' to the lesion, yet they lack similar structures and metal ion requirements. Here, we determined and analyzed crystal structures of a 2.4 ? resolution APE1-DNA product complex with Mg(2+) and a 0.92 Nfo with three metal ions. Structural and biochemical comparisons of these two evolutionarily distinct enzymes characterize key APE1 catalytic residues that are potentially functionally similar to Nfo active site components, as further tested and supported by computational analyses. We observe a magnesium-water cluster in the APE1 active site, with only Glu-96 forming the direct protein coordination to the Mg(2+). Despite differences in structure and metal requirements of APE1 and Nfo, comparison of their active site structures surprisingly reveals strong geometric conservation of the catalytic reaction, with APE1 catalytic side chains positioned analogously to Nfo metal positions, suggesting surprising functional equivalence between Nfo metal ions and APE1 residues. The finding that APE1 residues are positioned to substitute for Nfo metal ions is supported by the impact of mutations on activity. Collectively, the results illuminate the activities of residues, metal ions, and active site features for abasic site endonucleases.

  6. Size dependence of non-magnetic thickness in YIG nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Niyaifar, M., E-mail: md.niyaifar@gmail.com; Mohammadpour, H.; Dorafshani, M.; Hasanpour, A.

    2016-07-01

    This study is focused on particle size dependence of structural and magnetic properties in yttrium iron garnet (Y{sub 3}Fe{sub 5}O{sub 12}) nanoparticles. A series of YIG samples with different particle size were produced by varying the annealing temperatures. The X-ray analysis revealed an inverse correlation between lattice parameter and the crystallite size. The normal distribution is used for fitting the particles size distribution which is extracted from scanning electron micrographs. Also, by using the results of vibrating sample magnetometer, the magnetic diameter was calculated based on Langevin model in order to investigate the variation of dead layer thickness. Furthermore, the observed line broadening in Mössbauer spectra confirmed the increase of non-magnetic thickness due to the reduction of particle size. - Highlights: • Pure phase Y{sub 3}Fe{sub 5}O{sub 12} nanoparticles are fabricated in different particle size by a thermal treatment. • The size effect on magnetic properties is studied with a core/shell (magnetic/nonmagnetic) model. • The logarithmic variation of (dead layer thickness)/(particle size) ratio with the particle size is investigated. • The results of Mossbauer are explained based on the correlation between lattice constant and particle size variation.

  7. Murine leukemia virus pol gene products: analysis with antisera generated against reverse transcriptase and endonuclease fusion proteins expressed in Escherichia coli

    International Nuclear Information System (INIS)

    Hu, S.C.; Court, D.L.; Zweig, M.; Levin, J.G.

    1986-01-01

    The organization of the murine leukemia virus (MuLV) pol gene was investigated by expressing molecular clones containing AKR MuLV reverse transcriptase or endonuclease or both gene segments in Escherichia coli and generating specific antisera against the expressed bacterial proteins. Reaction of these antisera with detergent-disrupted virus precipitated and 80-kilodalton (kDa) protein, the MuLV reverse transcriptase, and a 46-kDa protein which we believe is the viral endonuclease. A third (50-kDa) protein, related to reverse transcriptase, was also precipitated. Bacterial extracts of clones expressing reverse transcriptase and endonuclease sequences competed with the viral 80- and 46-kDa proteins, respectively. These results demonstrate that the antisera are specific for viral reverse transcriptase and endonuclease. Immunoprecipitation of AKR MuLV with antisera prepared against a bacterial protein containing only endonuclease sequences led to the observation that reverse transcriptase and endonuclease can be associated as a complex involving a disulfide bond(s)

  8. Physical association of pyrimidine dimer DNA glycosylase and apurinic/apyrimidinic DNA endonuclease essential for repair of ultraviolet-damaged DNA

    International Nuclear Information System (INIS)

    Nakabeppu, Y.; Sekiguchi, M.

    1981-01-01

    T4 endonuclease, which is involved in repair of uv-damaged DNA, has been purified to apparent physical homogeneity. Incubation of uv-irradiated poly(dA).poly(dT) with the purified enzyme preparations resulted in production of alkali-labile apyrimidinic sites, followed by formation of nicks in the polymer. By performing a limited reaction with T4 endonuclease V at pH 8.5, irradiated polymer was converted to an intermediate form that carried a large number of alkali-labile sites but only a few nicks. The intermediate was used as substrate for the assay of apurinic/apyrimidinic DNA endonuclease activity. The two activities, a pyrimidine dimer DNA glycosylase and an apurinic/apyrimidinic DNA endonuclease, were copurified and found in enzyme preparations that contained only a 16,000-dalton polypeptide. These results strongly suggested that a DNA glycosylase specific for pyrimidine dimers and an apurinic/apyrimidinic DNA endonuclease reside in a single polypeptide chain coded by the denV gene of bacteriophage T4

  9. Modification of Bi:YIG film properties by substrate surface ion pre-treatment

    International Nuclear Information System (INIS)

    Shaposhnikov, A.N.; Prokopov, A.R.; Karavainikov, A.V.; Berzhansky, V.N.; Mikhailova, T.V.; Kotov, V.A.; Balabanov, D.E.; Sharay, I.V.; Salyuk, O.Y.; Vasiliev, M.; Golub, V.O.

    2014-01-01

    Highlights: • Effects of substrates ion beam treatment on magnetoptical properties Bi:YIG films. • Substrate surface damage results in sign inversion of the magneto-optical effects. • Atomically smooth films growth takes place on low energy ions treated substrates. • High energy ions treatment results in selective nucleation mechanism of the growth. - Abstract: The effect of a controlled ion beam pre-treatment of (1 1 1)-oriented Gd 3 Ga 5 O 12 substrates on the magneto-optical properties and surface morphology of the ultrathin bismuth-substituted yttrium–iron garnet films with a composition Bi 2.8 Y 0.2 Fe 5 O 12 was studied. It has been shown that the observed sign inversion of magneto-optical effects (Faraday rotation and magnetic circular dichroism) observed in films that were deposited on the GGG substrate pre-treated by 1 keV and 4 keV Ar + ion beams is a result of the substrate surface amorphization caused by the ion bombardment

  10. Biochemical properties and base excision repair complex formation of apurinic/apyrimidinic endonuclease from Pyrococcus furiosus

    OpenAIRE

    Kiyonari, Shinichi; Tahara, Saki; Shirai, Tsuyoshi; Iwai, Shigenori; Ishino, Sonoko; Ishino, Yoshizumi

    2009-01-01

    Apurinic/apyrimidinic (AP) sites are the most frequently found mutagenic lesions in DNA, and they arise mainly from spontaneous base loss or modified base removal by damage-specific DNA glycosylases. AP sites are cleaved by AP endonucleases, and the resultant gaps in the DNA are repaired by DNA polymerase/DNA ligase reactions. We identified the gene product that is responsible for the AP endonuclease activity in the hyperthermophilic euryarchaeon, Pyrococcus furiosus. Furthermore, we detected...

  11. Modulation of the DNA scanning activity of the Micrococcus luteus UV endonuclease

    International Nuclear Information System (INIS)

    Hamilton, R.W.; Lloyd, R.S.

    1989-01-01

    Micrococcus luteus UV endonuclease incises DNA at the sites of ultraviolet (UV) light-induced pyrimidine dimers. The mechanism of incision has been previously shown to be a glycosylic bond cleavage at the 5'-pyrimidine of the dimer followed by an apyrimidine endonuclease activity which cleaves the phosphodiester backbone between the pyrimidines. The process by which M. luteus UV endonuclease locates pyrimidine dimers within a population of UV-irradiated plasmids was shown to occur, in vitro, by a processive or sliding mechanism on non-target DNA as opposed to a distributive or random hit mechanism. Form I plasmid DNA containing 25 dimers per molecule was incubated with M. luteus UV endonuclease in time course reactions. The three topological forms of plasmid DNA generated were analyzed by agarose gel electrophoresis. When the enzyme encounters a pyrimidine dimer, it is significantly more likely to make only the glycosylase cleavage as opposed to making both the glycosylic and phosphodiester bond cleavages. Thus, plasmids are accumulated with many alkaline-labile sites relative to single-stranded breaks. In addition, reactions were performed at both pH 8.0 and pH 6.0, in the absence of NaCl, as well as 25,100, and 250 mM NaCl. The efficiency of the DNA scanning reaction was shown to be dependent on both the ionic strength and pH of the reaction. At low ionic strengths, the reaction was shown to proceed by a processive mechanism and shifted to a distributive mechanism as the ionic strength of the reaction increased. Processivity at pH 8.0 is shown to be more sensitive to increases in ionic strength than reactions performed at pH 6.0

  12. DNA scanning mechanism of T4 endonuclease V. Effect of NaCl concentration on processive nicking activity

    International Nuclear Information System (INIS)

    Gruskin, E.A.; Lloyd, R.S.

    1986-01-01

    T4 endonuclease V is a pyrimidine dimer-specific endonuclease which generates incisions in DNA at the sites of pyrimidine dimers by a processive reaction mechanism. A model is presented in which the degree of processivity is directly related to the efficacy of the one-dimensional diffusion of endonuclease V on DNA by which the enzyme locates pyrimidine dimers. The modulation of the processive nicking activity of T4 endonuclease V on superhelical covalently closed circular DNA (form I) which contains pyrimidine dimers has been investigated as a function of the ionic strength of the reaction. Agarose gel electrophoresis was used to separate the three topological forms of the DNA which were generated in time course reactions of endonuclease V with dimer-containing form I DNA in the absence of NaCl, and in 25, 50, and 100 mM NaCl. The degree of processivity was evaluated in terms of the mass fraction of form III (linear) DNA which was produced as a function of the fraction of form I DNA remaining. Processivity is maximal in the absence of NaCl and decreases as the NaCl concentration is increased. At 100 mM NaCl, processivity is abolished and endonuclease V generates incisions in DNA at the site of dimers by a distributive reaction mechanism. The change from the distributive to a processive reaction mechanism occurs at NaCl concentrations slightly below 50 mM. The high degree of processivity which is observed in the absence of NaCl is reversible to the distributive mechanism, as demonstrated by experiments in which the NaCl concentration was increased during the time course reaction. In addition, unirradiated DNA inhibited the incision of irradiated DNA only at NaCl concentrations at which processivity was observed

  13. [Restriction endonuclease digest - melting curve analysis: a new SNP genotyping and its application in traditional Chinese medicine authentication].

    Science.gov (United States)

    Jiang, Chao; Huang, Lu-Qi; Yuan, Yuan; Chen, Min; Hou, Jing-Yi; Wu, Zhi-Gang; Lin, Shu-Fang

    2014-04-01

    Single nucleotide polymorphisms (SNP) is an important molecular marker in traditional Chinese medicine research, and it is widely used in TCM authentication. The present study created a new genotyping method by combining restriction endonuclease digesting with melting curve analysis, which is a stable, rapid and easy doing SNP genotyping method. The new method analyzed SNP genotyping of two chloroplast SNP which was located in or out of the endonuclease recognition site, the results showed that when attaching a 14 bp GC-clamp (cggcgggagggcgg) to 5' end of the primer and selecting suited endonuclease to digest the amplification products, the melting curve of Lonicera japonica and Atractylodes macrocephala were all of double peaks and the adulterants Shan-yin-hua and A. lancea were of single peaks. The results indicated that the method had good stability and reproducibility for identifying authentic medicines from its adulterants. It is a potential SNP genotyping method and named restriction endonuclease digest - melting curve analysis.

  14. Modification of Bi:YIG film properties by substrate surface ion pre-treatment

    Energy Technology Data Exchange (ETDEWEB)

    Shaposhnikov, A.N.; Prokopov, A.R.; Karavainikov, A.V.; Berzhansky, V.N.; Mikhailova, T.V. [Taurida National V.I. Vernadsky University, Vernadsky Avenue, 4, Simferopol, 95007 (Ukraine); Kotov, V.A. [V.A. Kotelnikov Institute of Radio Engineering and Electronics, RAS, 11 Mohovaya Street, Moscow, 125009 (Russian Federation); Balabanov, D.E. [Moscow Institute of Physics and Technology, Dolgoprudny, 141700 (Russian Federation); Sharay, I.V.; Salyuk, O.Y. [Institute of Magnetism, NAS of Ukraine, 03142, Kiev (Ukraine); Vasiliev, M. [Electron Science Research Institute, Edith Cowan University, 270 Joondalup Drive, Joondalup 6027 (Australia); Golub, V.O., E-mail: v_o_golub@yahoo.com [Institute of Magnetism, NAS of Ukraine, 03142, Kiev (Ukraine)

    2014-07-01

    Highlights: • Effects of substrates ion beam treatment on magnetoptical properties Bi:YIG films. • Substrate surface damage results in sign inversion of the magneto-optical effects. • Atomically smooth films growth takes place on low energy ions treated substrates. • High energy ions treatment results in selective nucleation mechanism of the growth. - Abstract: The effect of a controlled ion beam pre-treatment of (1 1 1)-oriented Gd{sub 3}Ga{sub 5}O{sub 12} substrates on the magneto-optical properties and surface morphology of the ultrathin bismuth-substituted yttrium–iron garnet films with a composition Bi{sub 2.8}Y{sub 0.2}Fe{sub 5}O{sub 12} was studied. It has been shown that the observed sign inversion of magneto-optical effects (Faraday rotation and magnetic circular dichroism) observed in films that were deposited on the GGG substrate pre-treated by 1 keV and 4 keV Ar{sup +} ion beams is a result of the substrate surface amorphization caused by the ion bombardment.

  15. Presence of UV-endonuclease sensitive sites in daughter DNA of UV-irradiated mammalian cells

    International Nuclear Information System (INIS)

    D'Ambrosio, S.; Setlow, R.B.

    1978-02-01

    Asynchronous Chinese hamster cells were irradiated with 10 Jm -2 uv radiation and 0.25 to 4 hours later pulse-labeled with [ 3 H]thymidine. Cells synchronized by shaking off mitotic and G 1 cells were irradiated in either the G 1 -phase or S-phase of the cell cycle and pulse-labeled with [ 3 H]thymidine in the S-phase. After a 12 to 14 hour chase in unlabeled medium, the DNA was extracted, incubated with Micrococcus luteus uv-endonuclease and sedimented in alkaline sucrose. The number of endonuclease sensitive sites decreased as the time between uv irradiation and pulse-labeling of daughter DNA increased. Further, there were significantly less endonuclease sensitive sites in the daughter DNA from cells irradiated in the G 1 -phase than in the S-phase. These data indicate that very few, if any, dimers are transferred from parental DNA to daughter DNA and that the dimers detected in daughter DNA may be due to the irradiation of replicating daughter DNA before labeling

  16. Specific action of T4 endonuclease V on damaged DNA in xeroderma pigmentosum cells in vivo

    International Nuclear Information System (INIS)

    Tanaka, K.; Hayakawa, H.; Sekiguchi, M.; Okada, Y.

    1977-01-01

    The specific action of T4 endonuclease V on damaged DNA in xeroderma pigmentosum cells was examined using an in vivo assay system with hemagglutinating virus of Japan (Sendai virus) inactivated by uv light. A clear dose response was observed between the level of uv-induced unscheduled DNA synthesis of xeroderma pigmentosum cells and the amount of T4 endonuclease V activity added. The T4 enzyme was unstable in human cells, and its half-life was 3 hr. Fractions derived from an extract of Escherichia coli infected with T4v 1 , a mutant defective in the endonuclease V gene, showed no ability to restore the uv-induced unscheduled DNA synthesis of xeroderma pigmentosum cells. However, fractions derived from an extract of T4D-infected E. coli with endonuclease V activity were effective. The T4 enzyme was effective in xeroderma pigmentosum cells on DNA damaged by uv light but not in cells damaged by 4-nitroquinoline 1-oxide. The results of these experiments show that the T4 enzyme has a specific action on human cell DNA in vivo. Treatment with the T4 enzyme increased the survival of group A xeroderma pigmentosum cells after uv irradiation

  17. Molecular Recognition of DNA Damage Sites by Apurinic/Apyrimidinic Endonucleases

    Energy Technology Data Exchange (ETDEWEB)

    Braun, W. A.

    2005-07-28

    The DNA repair/redox factor AP endonuclease 1 (APE1) is a multifunctional protein which is known to to be essential for DNA repair activity in human cells. Structural/functional analyses of the APE activity is thus been an important research field to assess cellular defense mechanisms against ionizing radiation.

  18. Expression and Purification of BmrI Restriction Endonuclease and Its N-terminal Cleavage Domain Variants

    OpenAIRE

    Bao, Yongming; Higgins, Lauren; Zhang, Penghua; Chan, Siu-hong; Laget, Sophie; Sweeney, Suzanne; Lunnen, Keith; Xu, Shuang-yong

    2007-01-01

    BmrI (ACTGGG N5/N4) is one of the few metal-independent restriction endonucleases (REases) found in bacteria. The BmrI restriction-modification system was cloned by the methylase selection method, inverse PCR, and PCR. BmrI REase shows significant amino acid sequence identity to BfiI and a putative endonuclease MspBNCORF3798 from the sequenced Mesorhizobium sp. BNC1 genome. The EDTA-resistant BmrI REase was successfully over-expressed in a pre-modified E. coli strain from pET21a or pBAC-expIQ...

  19. Crystal structure and DNA-binding property of the ATPase domain of bacterial mismatch repair endonuclease MutL from Aquifex aeolicus.

    Science.gov (United States)

    Fukui, Kenji; Iino, Hitoshi; Baba, Seiki; Kumasaka, Takashi; Kuramitsu, Seiki; Yano, Takato

    2017-09-01

    DNA mismatch repair (MMR) system corrects mismatched bases that are generated mainly by DNA replication errors. The repair system excises the error-containing single-stranded region and enables the re-synthesis of the strand. In the early reactions of MMR, MutL endonuclease incises the newly-synthesized/error-containing strand of the duplex to initiate the downstream excision reaction. MutL endonuclease consists of the N-terminal ATPase and C-terminal endonuclease domains. In this study, we report the crystal structure of the ATPase domain of MutL endonuclease from Aquifex aeolicus. The overall structure of the domain was similar to those of human MutL homologs and Escherichia coli MutL, although E. coli MutL has no endonuclease activity. The ATPase domain was comprised of two subdomains: the N-terminal ATP-binding subdomain and the C-terminal α-β sandwich subdomain. Site-directed mutagenesis experiment identified DNA-interacting eight basic amino acid residues, which were distributed across both the two subdomains and formed a DNA-binding cleft. Docking simulation between the structures of the ATPase and endonuclease domains generated a reliable model structure for the full-length A. aeolicus MutL, which satisfies our previous result of small-angle X-ray scattering analysis. On the basis of the model structure and further experimental results, we concluded that the two separate DNA-binding sites in the full-length A. aeolicus MutL simultaneously bind a dsDNA molecule. Copyright © 2017 Elsevier B.V. All rights reserved.

  20. Influence of annealing temperature on structural and magnetic properties of pulsed laser-deposited YIG films on SiO2 substrate

    Science.gov (United States)

    Nag, Jadupati; Ray, Nirat

    2018-05-01

    Yttrium Iron Garnet (Y3Fe5O12) was synthesized by solid state/ceramic process. Thin films of YIG were deposited on SiO2 substrate at room temperature(RT) and at substrate temperature (Ts) 700 °C using pulsed laser deposition (PLD) technique. RT deposited thin films are amorphous in nature and non-magnetic. After annealing at temperature 800 ° RT deposited thin films showed X-ray peaks as well as the magnetic order. Magnetic ordering is enhanced by annealing temperature(Ta ≥ 750 °C) and resulted good quality of films with high magnetization value.

  1. Purification, crystallization, X-ray diffraction analysis and phasing of an engineered single-chain PvuII restriction endonuclease

    International Nuclear Information System (INIS)

    Meramveliotaki, Chrysi; Kotsifaki, Dina; Androulaki, Maria; Hountas, Athanasios; Eliopoulos, Elias; Kokkinidis, Michael

    2007-01-01

    PvuII is the first type II restriction endonuclease to be converted from its wild-type homodimeric form into an enzymatically active single-chain variant. The enzyme was crystallized and phasing was successfully performed by molecular replacement. The restriction endonuclease PvuII from Proteus vulgaris has been converted from its wild-type homodimeric form into the enzymatically active single-chain variant scPvuII by tandemly joining the two subunits through the peptide linker Gly-Ser-Gly-Gly. scPvuII, which is suitable for the development of programmed restriction endonucleases for highly specific DNA cleavage, was purified and crystallized. The crystals diffract to a resolution of 2.35 Å and belong to space group P4 2 , with unit-cell parameters a = b = 101.92, c = 100.28 Å and two molecules per asymmetric unit. Phasing was successfully performed by molecular replacement

  2. Measurement of M. luteus endonuclease-sensitive lesions by alkaline elution

    Energy Technology Data Exchange (ETDEWEB)

    Fornace, Jr, A J [National Cancer Inst., Bethesda, MD (USA). Lab. for Experimental Pathology

    1982-01-01

    The UV-endonuclease approach to detect DNA damage has been combined with the alkaline elution technique with a resultant marked increase in sensitivity compared to the conventional method using alkaline sedimentation. DNA from UV-irradiated cells was digested on an inert filter with an extract from Micrococcus luteus and then analyzed by alkaline elution. Endonuclease-sensitive sites (endo-sites) were measured after doses of 0.08-0.7 Jm/sup -2/ of UV-radiation. An estimate of endo-site production with UV radiation, 0.27 endo-sites/10/sup 8/ daltons of DNA/0.1 Jm/sup -2/, was similar to that usually seen at higher doses by others. With repair incubation, approx. 50% of the endo-sites were removed in 4 h by normal human fibroblasts after 0.2 or 0.4 Jm/sup -2/, no appreciable repair was seen in xeroderma pigmentosum fibroblasts from complementation group A after 24 h of repair incubation. No photoreaction of UV damage due to 0.4 Jm/sup -2/ was detected in normal human fibroblasts.

  3. Measurement of M. luteus endonuclease-sensitive lesions by alkaline elution

    International Nuclear Information System (INIS)

    Fornace, A.J. Jr.

    1982-01-01

    The UV-endonuclease approach to detect DNA damage has been combined with the alkaline elution technique with a resultant marked increase in sensitivity compared to the conventional method using alkaline sedimentation. DNA from UV-irradiated cells was digested on an inert filter with an extract from Micrococcus luteus and then analyzed by alkaline elution. Endonuclease-sensitive sites (endo-sites) were measured after doses of 0.08-0.7 Jm -2 of UV-radiation. An estimate of endo-site production with UV radiation, 0.27 endo-sites/10 8 daltons of DNA/0.1 Jm -2 , was similar to that usually seen at higher doses by others. With repair incubation, approx. 50% of the endo-sites were removed in 4 h by normal human fibroblasts after 0.2 or 0.4 Jm -2 , no appreciable repair was seen in xeroderma pigmentosum fibroblasts from complementation group A after 24 h of repair incubation. No photoreaction of UV damage due to 0.4 Jm -2 was detected in normal human fibroblasts. (orig./AJ)

  4. The mismatch repair and meiotic recombination endonuclease Mlh1-Mlh3 is activated by polymer formation and can cleave DNA substrates in trans.

    Science.gov (United States)

    Manhart, Carol M; Ni, Xiaodan; White, Martin A; Ortega, Joaquin; Surtees, Jennifer A; Alani, Eric

    2017-04-01

    Crossing over between homologs is initiated in meiotic prophase by the formation of DNA double-strand breaks that occur throughout the genome. In the major interference-responsive crossover pathway in baker's yeast, these breaks are resected to form 3' single-strand tails that participate in a homology search, ultimately forming double Holliday junctions (dHJs) that primarily include both homologs. These dHJs are resolved by endonuclease activity to form exclusively crossovers, which are critical for proper homolog segregation in Meiosis I. Recent genetic, biochemical, and molecular studies in yeast are consistent with the hypothesis of Mlh1-Mlh3 DNA mismatch repair complex acting as the major endonuclease activity that resolves dHJs into crossovers. However, the mechanism by which the Mlh1-Mlh3 endonuclease is activated is unknown. Here, we provide evidence that Mlh1-Mlh3 does not behave like a structure-specific endonuclease but forms polymers required to generate nicks in DNA. This conclusion is supported by DNA binding studies performed with different-sized substrates that contain or lack polymerization barriers and endonuclease assays performed with varying ratios of endonuclease-deficient and endonuclease-proficient Mlh1-Mlh3. In addition, Mlh1-Mlh3 can generate religatable double-strand breaks and form an active nucleoprotein complex that can nick DNA substrates in trans. Together these observations argue that Mlh1-Mlh3 may not act like a canonical, RuvC-like Holliday junction resolvase and support a novel model in which Mlh1-Mlh3 is loaded onto DNA to form an activated polymer that cleaves DNA.

  5. Yeast redoxyendonuclease, a DNA repair enzyme similar to Escherichia coli endonuclease III

    International Nuclear Information System (INIS)

    Gossett, J.; Lee, K.; Cunningham, R.P.; Doetsch, P.W.

    1988-01-01

    A DNA repair endonuclease (redoxyendonuclease) was isolated from bakers' yeast (Saccharomyces cerevisiae). The enzyme has been purified by a series of column chromatography steps and cleaves OsO 4 -damaged, double-stranded DNA at sites of thymine glycol and heavily UV-irradiated DNA at sites of cytosine, thymine, and guanine photoproducts. The base specificity and mechanism of phosphodiester bond cleavage for the yeast redoxyendonuclease appear to be identical with those of Escherichia coli endonuclease III when thymine glycol containing, end-labeled DNA fragments of defined sequence are employed as substrates. Yeast redoxyendonuclease has an apparent molecular size of 38,000-42,000 daltons and is active in the absence of divalent metal cations. The identification of such an enzyme in yeast may be of value in the elucidation of the biochemical basis for radiation sensitivity in certain yeast mutants

  6. Inroads into base excision repair I. The discovery of apurinic/apyrimidinic (AP) endonuclease. "An endonuclease for depurinated DNA in Escherichia coli B," Canadian Journal of Biochemistry, 1972.

    Science.gov (United States)

    Lindahl, Tomas; Verly, W G; Paquette Y

    2004-11-02

    DNA treated with alkylating agents is incised at sites of damage by cell extracts. A key component of this DNA repair function was shown by Verly and co-workers to be an endonuclease acting at secondary lesions, apurinic sites, rather than directly at alkylated nucleotide residues.

  7. Molecular mechanism of VDE-initiated intein homing in yeast nuclear genome.

    Science.gov (United States)

    Fukuda, Tomoyuki; Nagai, Yuri; Ohya, Yoshikazu

    2004-01-01

    In Saccharomyces cerevisiae, VMA1 intein encodes a homing endonuclease termed VDE which is produced by an autocatalytic protein splicing reaction. VDE introduces a DSB at its recognition sequence on intein-minus allele, resulting in the lateral transfer of VMA1 intein. In this review, we summarize a decade of in vitro study on VDE and describe our recent study on the in vivo behavior of both VDE and host proteins involved in intein mobility. Meiotic DSBs caused by VDE are repaired in the similar pathway to that working in meiotic recombination induced by Spo11p-mediated DSBs. Meiosis-specific DNA cleavage and homing is shown to be guaranteed by the two distinct mechanisms, the subcellular localization of VDE and a requirement of premeiotic DNA replication. Based on these lines of evidence, we present the whole picture of molecular mechanism of VDE-initiated homing in yeast cells.

  8. Enhancement of ultraviolet-DNA repair in denV gene transfectants and T4 endonuclease V-liposome recipients

    International Nuclear Information System (INIS)

    Kibitel, J.T.; Yee, V.; Yarosh, D.B.

    1991-01-01

    The phage T4 denV gene, coding for the pyrimidine-dimer specific T4 endonuclease V, was transfected into human repair-proficient fibroblasts, repair-deficient xeroderma pigmentosum fibroblasts, and wild type CHO hamster cells. Transfectants maintained denV DNA and expressed denV mRNA. Purified T4 endonuclease V encapsulated in liposomes was also used to treat repair-proficient and -deficient human cells. The denV transfected clones and liposome-treated cells showed increased unscheduled DNA synthesis and enhanced removal of pyrimidine dimers compared to controls. Both denV gene transfection and endonuclease V liposome treatment enhanced post-UV survival in xeroderma pigmentosum cells but had no effect on survival in repair-proficient human or hamster cells. The results demonstrate that an exogenous DNA repair enzyme can correct the DNA repair defect in xeroderma pigmentosum cells and enhance DNA repair in normal cells. (author)

  9. Atomic Structure and Biochemical Characterization of an RNA Endonuclease in the N Terminus of Andes Virus L Protein.

    Directory of Open Access Journals (Sweden)

    Yaiza Fernández-García

    2016-06-01

    Full Text Available Andes virus (ANDV is a human-pathogenic hantavirus. Hantaviruses presumably initiate their mRNA synthesis by using cap structures derived from host cell mRNAs, a mechanism called cap-snatching. A signature for a cap-snatching endonuclease is present in the N terminus of hantavirus L proteins. In this study, we aimed to solve the atomic structure of the ANDV endonuclease and characterize its biochemical features. However, the wild-type protein was refractory to expression in Escherichia coli, presumably due to toxic enzyme activity. To circumvent this problem, we introduced attenuating mutations in the domain that were previously shown to enhance L protein expression in mammalian cells. Using this approach, 13 mutant proteins encompassing ANDV L protein residues 1-200 were successfully expressed and purified. Protein stability and nuclease activity of the mutants was analyzed and the crystal structure of one mutant was solved to a resolution of 2.4 Å. Shape in solution was determined by small angle X-ray scattering. The ANDV endonuclease showed structural similarities to related enzymes of orthobunya-, arena-, and orthomyxoviruses, but also differences such as elongated shape and positively charged patches surrounding the active site. The enzyme was dependent on manganese, which is bound to the active site, most efficiently cleaved single-stranded RNA substrates, did not cleave DNA, and could be inhibited by known endonuclease inhibitors. The atomic structure in conjunction with stability and activity data for the 13 mutant enzymes facilitated inference of structure-function relationships in the protein. In conclusion, we solved the structure of a hantavirus cap-snatching endonuclease, elucidated its catalytic properties, and present a highly active mutant form, which allows for inhibitor screening.

  10. Cofactor requirement of HpyAV restriction endonuclease.

    Directory of Open Access Journals (Sweden)

    Siu-Hong Chan

    Full Text Available BACKGROUND: Helicobacter pylori is the etiologic agent of common gastritis and a risk factor for gastric cancer. It is also one of the richest sources of Type II restriction-modification (R-M systems in microorganisms. PRINCIPAL FINDINGS: We have cloned, expressed and purified a new restriction endonuclease HpyAV from H. pylori strain 26695. We determined the HpyAV DNA recognition sequence and cleavage site as CCTTC 6/5. In addition, we found that HpyAV has a unique metal ion requirement: its cleavage activity is higher with transition metal ions than in Mg(++. The special metal ion requirement of HpyAV can be attributed to the presence of a HNH catalytic site similar to ColE9 nuclease instead of the canonical PD-X-D/EXK catalytic site found in many other REases. Site-directed mutagenesis was carried out to verify the catalytic residues of HpyAV. Mutation of the conserved metal-binding Asn311 and His320 to alanine eliminated cleavage activity. HpyAV variant H295A displayed approximately 1% of wt activity. CONCLUSIONS/SIGNIFICANCE: Some HNH-type endonucleases have unique metal ion cofactor requirement for optimal activities. Homology modeling and site-directed mutagenesis confirmed that HpyAV is a member of the HNH nuclease family. The identification of catalytic residues in HpyAV paved the way for further engineering of the metal binding site. A survey of sequenced microbial genomes uncovered 10 putative R-M systems that show high sequence similarity to the HpyAV system, suggesting lateral transfer of a prototypic HpyAV-like R-M system among these microorganisms.

  11. Identification of a mismatch-specific endonuclease in hyperthermophilic Archaea

    OpenAIRE

    Ishino, Sonoko; Nishi, Yuki; Oda, Soichiro; Uemori, Takashi; Sagara, Takehiro; Takatsu, Nariaki; Yamagami, Takeshi; Shirai, Tsuyoshi; Ishino, Yoshizumi

    2016-01-01

    The common mismatch repair system processed by MutS and MutL and their homologs was identified in Bacteria and Eukarya. However, no evidence of a functional MutS/L homolog has been reported for archaeal organisms, and it is not known whether the mismatch repair system is conserved in Archaea. Here, we describe an endonuclease that cleaves double-stranded DNA containing a mismatched base pair, from the hyperthermophilic archaeon Pyrococcus furiosus. The corresponding gene revealed that the act...

  12. Cleavage and protection of locked nucleic acid-modified DNA by restriction endonucleases

    DEFF Research Database (Denmark)

    Crouzier, Lucile; Dubois, Camille; Wengel, Jesper

    2012-01-01

    Locked nucleic acid (LNA) is one of the most prominent nucleic acid analogues reported so far. We herein for the first time report cleavage by restriction endonuclease of LNA-modified DNA oligonucleotides. The experiments revealed that RsaI is an efficient enzyme capable of recognizing and cleaving...

  13. Comparison of the cleavage of pyrimidine dimers by the bacteriophage T4 and Micrococcus luteus uv-specific endonucleases

    International Nuclear Information System (INIS)

    Gordon, L.K.; Haseltine, W.A.

    1980-01-01

    A comparison was made of the activity of the uv-specific endonucleases of bacteriophage T4 (T4 endonuclease V) and of Micrococcus luteus on ultraviolet light-irradiated DNA substrates of defined sequence. The two enzyms cleave DNA at the site of pyrimidine dimers with the same frequency. The products of the cleavage reaction are the same. The pyrimidine dimer DNA-glycosylase activity of both enzymes is more active on double-stranded DNA than it is on single-stranded DNA

  14. Recruitment of RecA homologs Dmc1p and Rad51p to the double-strand break repair site initiated by meiosis-specific endonuclease VDE (PI-SceI).

    Science.gov (United States)

    Fukuda, Tomoyuki; Ohya, Yoshikazu

    2006-02-01

    During meiosis, VDE (PI-SceI), a homing endonuclease in Saccharomyces cerevisiae, introduces a double-strand break (DSB) at its recognition sequence and induces homologous recombinational repair, called homing. Meiosis-specific RecA homolog Dmc1p, as well as mitotic RecA homolog Rad51p, acts in the process of meiotic recombination, being required for strand invasion and exchange. In this study, recruitment of Dmc1p and Rad51p to the VDE-induced DSB repair site is investigated by chromatin immunoprecipitation assay. It is revealed that Dmc1p and Rad51p are loaded to the repair site in an independent manner. Association of Rad51p requires other DSB repair proteins of Rad52p, Rad55p, and Rad57p, while loading of Dmc1p is facilitated by the different protein, Sae3p. Absence of Tid1p, which can bind both RecA homologs, appears specifically to cause an abnormal distribution of Dmc1p. Lack of Hop2, Mnd1p, and Sae1p does not impair recruitment of both RecA homologs. These findings reveal the discrete functions of each strand invasion protein in VDE-initiated homing, confirm the similarity between VDE-initiated homing and Spo11p-initiated meiotic recombination, and demonstrate the availability of VDE-initiated homing for the study of meiotic recombination.

  15. Type II restriction endonucleases : a historical perspective and more

    OpenAIRE

    Pingoud, Alfred; Wilson, Geoffrey G.; Wende, Wolfgang

    2014-01-01

    This article continues the series of Surveys and Summaries on restriction endonucleases (REases) begun this year in Nucleic Acids Research. Here we discuss ‘Type II’ REases, the kind used for DNA analysis and cloning. We focus on their biochemistry: what they are, what they do, and how they do it. Type II REases are produced by prokaryotes to combat bacteriophages. With extreme accuracy, each recognizes a particular sequence in double-stranded DNA and cleaves at a fixed position within or nea...

  16. Cell-Autonomous Progeroid Changes in Conditional Mouse Models for Repair Endonuclease XPG Deficiency

    NARCIS (Netherlands)

    S. Barnhoorn (Sander); L.M. Uittenboogaard (Lieneke); D. Jaarsma (Dick); W.P. Vermeij (Wilbert); M. Tresini (Maria); M. Weymaere (Michael); H. Menoni (Hervé); R.M.C. Brandt (Renata); M.C. de Waard (Monique); S.M. Botter (Sander); A.H. Sarker (Altraf); N.G.J. Jaspers (Nicolaas); G.T.J. van der Horst (Gijsbertus); P.K. Cooper (Priscilla K.); J.H.J. Hoeijmakers (Jan); I. van der Pluijm (Ingrid)

    2014-01-01

    textabstractAs part of the Nucleotide Excision Repair (NER) process, the endonuclease XPG is involved in repair of helix-distorting DNA lesions, but the protein has also been implicated in several other DNA repair systems, complicating genotype-phenotype relationship in XPG patients. Defects in XPG

  17. Structure of the I-SceI nuclease complexed with its dsDNA target and three catalytic metal ions

    DEFF Research Database (Denmark)

    Prieto, Jesús; Redondo, Pilar; Merino, Nekane

    2016-01-01

    Homing endonucleases are highly specific DNA-cleaving enzymes that recognize and cleave long stretches of DNA. The engineering of these enzymes provides instruments for genome modification in a wide range of fields, including gene targeting. The homing endonuclease I-SceI from the yeast Saccharom......Homing endonucleases are highly specific DNA-cleaving enzymes that recognize and cleave long stretches of DNA. The engineering of these enzymes provides instruments for genome modification in a wide range of fields, including gene targeting. The homing endonuclease I-SceI from the yeast...... experiments were performed in the presence of Mn(2+), yielding crystals that were suitable for X-ray diffraction analysis. The crystals belonged to the orthorhombic space group P212121, with unit-cell parameters a = 80.11, b = 80.57, c = 130.87 Å, α = β = γ = 90°. The self-rotation function and the Matthews...

  18. An analysis of the repair processes in ultraviolet-irradiated Micrococcus luteus using purified ultraviolet-endonuclease

    International Nuclear Information System (INIS)

    Tomilin, N.V.; Zherebtsov, S.V.

    1982-01-01

    The measurement of the frequency of endonucleolytic incisions in ultraviolet-irradiated DNA serves as the test for the presence of pyrimidine dimers. In accordance with this approach, the lysates of three Micrococcus luteus strains containing radioactively labeled chromosomes were treated with purified M. luteus ultraviolet-endonuclease to trace segregation of dimers amongst parental and newly synthesized DNA and their removal during postreplication and excision DNA repair. A considerable proportion of the dimers in all strains tested proved to be insensitive to the action of exogenous incising enzyme. The use of chloramphenicol as an inhibitor of postirradiation protein synthesis in combination with ultraviolet-endonuclease treatment of DNA allowed to reveal at least two alternative pathways of postreplication repair: constitutively active recombinational pathway and inducible nonrecombinational one. (Auth.)

  19. A detailed experimental study of a DNA computer with two endonucleases.

    Science.gov (United States)

    Sakowski, Sebastian; Krasiński, Tadeusz; Sarnik, Joanna; Blasiak, Janusz; Waldmajer, Jacek; Poplawski, Tomasz

    2017-07-14

    Great advances in biotechnology have allowed the construction of a computer from DNA. One of the proposed solutions is a biomolecular finite automaton, a simple two-state DNA computer without memory, which was presented by Ehud Shapiro's group at the Weizmann Institute of Science. The main problem with this computer, in which biomolecules carry out logical operations, is its complexity - increasing the number of states of biomolecular automata. In this study, we constructed (in laboratory conditions) a six-state DNA computer that uses two endonucleases (e.g. AcuI and BbvI) and a ligase. We have presented a detailed experimental verification of its feasibility. We described the effect of the number of states, the length of input data, and the nondeterminism on the computing process. We also tested different automata (with three, four, and six states) running on various accepted input words of different lengths such as ab, aab, aaab, ababa, and of an unaccepted word ba. Moreover, this article presents the reaction optimization and the methods of eliminating certain biochemical problems occurring in the implementation of a biomolecular DNA automaton based on two endonucleases.

  20. Survival of Saccharomyces cerevisiae after treatment with the restriction endonuclease Alu I

    International Nuclear Information System (INIS)

    Winckler, K.; Bach, B.; Obe, G.

    1988-01-01

    Treatment of yeast cells proficient in the repair of radiation damage (Saccharomyces cervisiae) with the restriction endonuclease Alu I leads to a positive dose-effect relationship between inactivation level and enzyme concentration. The data suggest an uptake of the active restriction enzyme into the cells and a relationship between induction of DNA double-strand breaks and cell killing. (author)

  1. Endonuclease G is a novel determinant of cardiac hypertrophy and mitochondrial function

    Czech Academy of Sciences Publication Activity Database

    McDermott-Roe, Ch.; Ye, J.; Ahmed, R.; Sun, X. M.; Serafín, A.; Ware, J.; Bottolo, L.; Muckett, P.; Caňas, X.; Zhang, J.; Rowe, G. C.; Buchan, R.; Lu, H.; Braithwaite, A.; Mancini, M.; Hauton, D.; Martí, R.; García-Arumí, E.; Hubner, N.; Jacob, H.; Serikawa, T.; Zídek, Václav; Papoušek, František; Kolář, František; Cardona, M.; Ruiz-Meana, M.; García-Dorado, D.; Comella, J. X.; Felkin, L. E.; Barton, P. J. R.; Arany, Z.; Pravenec, Michal; Petretto, E.; Sanchis, D.; Cook, S.A.

    2011-01-01

    Roč. 478, č. 7367 (2011), s. 114-118 ISSN 0028-0836 R&D Projects: GA MŠk(CZ) 1M0520; GA ČR(CZ) GA301/08/0166 Institutional research plan: CEZ:AV0Z50110509 Keywords : left ventricular hypertrophy * endonuclease G * mitochondrial dysfunction Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 36.280, year: 2011

  2. Transient and Switchable (Triethylsilyl)ethynyl Protection of DNA against Cleavage by Restriction Endonucleases

    Czech Academy of Sciences Publication Activity Database

    Kielkowski, Pavel; Macíčková-Cahová, Hana; Pohl, Radek; Hocek, Michal

    2011-01-01

    Roč. 50, č. 37 (2011), s. 8727-8730 ISSN 1433-7851 R&D Projects: GA ČR GA203/09/0317 Institutional research plan: CEZ:AV0Z40550506 Keywords : alkynes * DNA * protecting groups * nucleotides * restriction endonucleases Subject RIV: CC - Organic Chemistry Impact factor: 13.455, year: 2011

  3. Human RECQL5beta stimulates flap endonuclease 1

    DEFF Research Database (Denmark)

    Speina, Elzbieta; Dawut, Lale; Hedayati, Mohammad

    2010-01-01

    devoid of RECQL1 and RECQL5 display increased chromosomal instability. Here, we report the physical and functional interaction of the large isomer of RECQL5, RECQL5beta, with the human flap endonuclease 1, FEN1, which plays a critical role in DNA replication, recombination and repair. RECQL5beta...... dramatically stimulates the rate of FEN1 cleavage of flap DNA substrates. Moreover, we show that RECQL5beta and FEN1 interact physically and co-localize in the nucleus in response to DNA damage. Our findings, together with the previous literature on WRN, BLM and RECQL4's stimulation of FEN1, suggests...

  4. Biological significance of facilitated diffusion in protein-DNA interactions. Applications to T4 endonuclease V-initiated DNA repair

    International Nuclear Information System (INIS)

    Dowd, D.R.; Lloyd, R.S.

    1990-01-01

    Facilitated diffusion along nontarget DNA is employed by numerous DNA-interactive proteins to locate specific targets. Until now, the biological significance of DNA scanning has remained elusive. T4 endonuclease V is a DNA repair enzyme which scans nontarget DNA and processively incises DNA at the site of pyrimidine dimers which are produced by exposure to ultraviolet (UV) light. In this study we tested the hypothesis that there exists a direct correlation between the degree of processivity of wild type and mutant endonuclease V molecules and the degree of enhanced UV resistance which is conferred to repair-deficient Eshcerichia coli. This was accomplished by first creating a series of endonuclease V mutants whose in vitro catalytic activities were shown to be very similar to that of the wild type enzyme. However, when the mechanisms by which these enzymes search nontarget DNA for its substrate were analyzed in vitro and in vivo, the mutants displayed varying degrees of nontarget DNA scanning ranging from being nearly as processive as wild type to randomly incising dimers within the DNA population. The ability of these altered endonuclease V molecules to enhance UV survival in DNA repair-deficient E. coli then was assessed. The degree of enhanced UV survival was directly correlated with the level of facilitated diffusion. This is the first conclusive evidence directly relating a reduction of in vivo facilitated diffusion with a change in an observed phenotype. These results support the assertion that the mechanisms which DNA-interactive proteins employ in locating their target sites are of biological significance

  5. Restriction endonuclease analysis of chloroplast DNA in interspecies somatic Hybrids of Petunia.

    Science.gov (United States)

    Kumar, A; Cocking, E C; Bovenberg, W A; Kool, A J

    1982-12-01

    Restriction endonuclease cleavage pattern analysis of chloroplast DNA (cpDNA) of three different interspecific somatic hybrid plants revealed that the cytoplasms of the hybrids contained only cpDNA of P. parodii. The somatic hybrid plants analysed were those between P. parodii (wild type) + P. hybrida (wild type); P. parodii (wild type)+P. inflata (cytoplasmic albino mutant); P. parodii (wild type) + P. parviflora (nuclear albino mutant). The presence of only P. parodii chloroplasts in the somatic hybrid of P. parodii + P. inflata is possibly due to the stringent selection used for somatic hybrid production. However, in the case of the two other somatic hybrids P. parodii + P. hybrida and P. parodii + P. parviflora it was not possible to determine whether the presence of only P. parodii chloroplasts in these somatic hybrid plants was due to the nature of the selection schemes used or simply occurred by chance. The relevance of such somatic hybrid material for the study of genomic-cytoplasmic interaction is discussed, as well as the use of restriction endonuclease fragment patterns for the analysis of taxonomic and evolutionary inter-relationships in the genus Petunia.

  6. Cleavage of DNA containing 5-fluorocytosine or 5-fluorouracil by type II restriction endonucleases

    Czech Academy of Sciences Publication Activity Database

    Olszewska, Agata; Daďová, Jitka; Mačková, Michaela; Hocek, Michal

    2015-01-01

    Roč. 23, č. 21 (2015), s. 6885-6890 ISSN 0968-0896 R&D Projects: GA ČR GA14-04289S Institutional support: RVO:61388963 Keywords : modified nucleotides * DNA * restriction endonucleases * DNA polymerase * pyrimidine nucleosides Subject RIV: CC - Organic Chemistry Impact factor: 2.923, year: 2015

  7. Molecular dynamics simulations of deoxyribonucleic acids and repair enzyme T4 endonuclease V

    International Nuclear Information System (INIS)

    Pinak, Miroslav

    1999-01-01

    This report describes the results of molecular dynamics (MD) simulation of deoxyribonucleic acids (DNA) and specific repair enzyme T4 endonuclease V. Namely research described here is focused on the examination of specific recognition process, in which this repair enzyme recognizes the damaged site on the DNA molecule-thymine dimer (TD). TD is frequent DNA damage induced by UV radiation in sun light and unless properly repaired it may be mutagenic or lethal for cell, and is also considered among the major causes of skin cancer. T4 endonuclease V is a DNA specific repair enzyme from bacteriophage T4 that catalyzes the first reaction step of TD repair pathway. MD simulations of three molecules - native DNA dodecamer (12 base pairs), DNA of the same sequence of nucleotides as native one but with TD, and repair enzyme T4 endonuclease V - were performed for 1 ns individually for each molecule. Simulations were analyzed to determine the role of electrostatic interaction in the recognition process. It is found that electrostatic energies calculated for amino acids of the enzyme have positive values of around +15 kcal/mol. The electrostatic energy of TD site has negative value of approximately -9 kcal/mol, different from the nearly neutral value of the respective thymines site of the native DNA. The electrostatic interaction of TD site with surrounding water environment differs from the electrostatic interaction of other nucleotides. Differences found between TD site and respective thymines site of native DNA indicate that the electrostatic energy is an important factor contributing to proper recognition of TD site during scanning process in which enzyme scans the DNA. In addition to the electrostatic energy, the important factor in recognition process might be structural complementarity of enzyme and bent DNA with TD. There is significant kink formed around TD site, that is not observed in native DNA. (author)

  8. RPA activates the XPF‐ERCC1 endonuclease to initiate processing of DNA interstrand crosslinks

    KAUST Repository

    Abdullah, Ummi B

    2017-06-13

    During replication‐coupled DNA interstrand crosslink (ICL) repair, the XPF‐ERCC1 endonuclease is required for the incisions that release, or “unhook”, ICLs, but the mechanism of ICL unhooking remains largely unknown. Incisions are triggered when the nascent leading strand of a replication fork strikes the ICL. Here, we report that while purified XPF‐ERCC1 incises simple ICL‐containing model replication fork structures, the presence of a nascent leading strand, modelling the effects of replication arrest, inhibits this activity. Strikingly, the addition of the single‐stranded DNA (ssDNA)‐binding replication protein A (RPA) selectively restores XPF‐ERCC1 endonuclease activity on this structure. The 5′–3′ exonuclease SNM1A can load from the XPF‐ERCC1‐RPA‐induced incisions and digest past the crosslink to quantitatively complete the unhooking reaction. We postulate that these collaborative activities of XPF‐ERCC1, RPA and SNM1A might explain how ICL unhooking is achieved in vivo.

  9. RPA activates the XPF‐ERCC1 endonuclease to initiate processing of DNA interstrand crosslinks

    KAUST Repository

    Abdullah, Ummi B; McGouran, Joanna F; Brolih, Sanja; Ptchelkine, Denis; El‐Sagheer, Afaf H; Brown, Tom; McHugh, Peter J

    2017-01-01

    During replication‐coupled DNA interstrand crosslink (ICL) repair, the XPF‐ERCC1 endonuclease is required for the incisions that release, or “unhook”, ICLs, but the mechanism of ICL unhooking remains largely unknown. Incisions are triggered when the nascent leading strand of a replication fork strikes the ICL. Here, we report that while purified XPF‐ERCC1 incises simple ICL‐containing model replication fork structures, the presence of a nascent leading strand, modelling the effects of replication arrest, inhibits this activity. Strikingly, the addition of the single‐stranded DNA (ssDNA)‐binding replication protein A (RPA) selectively restores XPF‐ERCC1 endonuclease activity on this structure. The 5′–3′ exonuclease SNM1A can load from the XPF‐ERCC1‐RPA‐induced incisions and digest past the crosslink to quantitatively complete the unhooking reaction. We postulate that these collaborative activities of XPF‐ERCC1, RPA and SNM1A might explain how ICL unhooking is achieved in vivo.

  10. Permeabilization of ultraviolet-irradiated chinese hamster cells with polyethylene glycol and introduction of ultraviolet endonuclease from Micrococcus luteus

    International Nuclear Information System (INIS)

    Yarosh, D.B.; Setlow, R.B.

    1981-01-01

    Chinese hamster V-79 cells were made permeable by treatment with polyethylene glycol and then incubated with a Micrococcus luteus extract containing ultraviolet-specific endonuclease activity. This treatment introduced nicks in irradiated, but not in unirradiated, deoxyribonucleic acid. The nicks remained open for at least 3 h; there was no loss of endonuclease-sensitive sites, and no excision of dimers as measured by chromatography was detected. In addition, there was no increase in ultraviolet resistance in treated cells. This suggests that the absence of a significant amount of excision repair in rodent cells is due to the lack of both incision and excision capacity

  11. Structural studies on metal-containing enzymes: T4 endonuclease VII and D. gigas formate dehydrogenase

    NARCIS (Netherlands)

    Raaijmakers, H.C.A.

    2001-01-01

    Many biological processes require metal ions, and many of these metal-ion functions involve metalloproteins. The metal ions in metalloproteins are often critical to the protein's function, structure, or stability. This thesis focuses on two of these proteins, bacteriophage T4 endonuclease

  12. Mlh1-Mlh3, a Meiotic Crossover and DNA Mismatch Repair Factor, Is a Msh2-Msh3-stimulated Endonuclease*

    Science.gov (United States)

    Rogacheva, Maria V.; Manhart, Carol M.; Chen, Cheng; Guarne, Alba; Surtees, Jennifer; Alani, Eric

    2014-01-01

    Crossing over between homologous chromosomes is initiated in meiotic prophase in most sexually reproducing organisms by the appearance of programmed double strand breaks throughout the genome. In Saccharomyces cerevisiae the double-strand breaks are resected to form three prime single-strand tails that primarily invade complementary sequences in unbroken homologs. These invasion intermediates are converted into double Holliday junctions and then resolved into crossovers that facilitate homolog segregation during Meiosis I. Work in yeast suggests that Msh4-Msh5 stabilizes invasion intermediates and double Holliday junctions, which are resolved into crossovers in steps requiring Sgs1 helicase, Exo1, and a putative endonuclease activity encoded by the DNA mismatch repair factor Mlh1-Mlh3. We purified Mlh1-Mlh3 and showed that it is a metal-dependent and Msh2-Msh3-stimulated endonuclease that makes single-strand breaks in supercoiled DNA. These observations support a direct role for an Mlh1-Mlh3 endonuclease activity in resolving recombination intermediates and in DNA mismatch repair. PMID:24403070

  13. Selective inhibition by methoxyamine of the apurinic/apyrimidinic endonuclease activity associated with pyrimidine dimer-DNA glycosylases from Micrococcus luteus and bacteriophage T4

    International Nuclear Information System (INIS)

    Liuzzi, M.; Weinfeld, M.; Paterson, M.C.

    1987-01-01

    The UV endonucleases from Micrococcus luteus and bacteriophage T4 possess two catalytic activities specific for the site of cyclobutane pyrimidine dimers in UV-irradiated DNA: a DNA glycosylase that cleaves the 5'-glycosyl bond of the dimerized pyrimidines and an apurinic/apyrimidinic (AP) endonuclease that thereupon incises the phosphodiester bond 3' to the resulting apyrimidinic site. The authors have explored the potential use of methoxyamine, a chemical that reacts at neutral pH with AP sites in DNA, as a selective inhibitor of the AP endonuclease activities residing in the M. luteus and T4 enzymes. The presence of 50 mM methoxyamine during incubation of UV-treated, [ 3 H]thymine-labeled poly(dA) x poly(dT) with either enzyme preparation was found to protect completely the irradiated copolymer from endonucleolytic attack at dimer sites, as assayed by yield of acid-soluble radioactivity. In contrast, the dimer-DNA glycosylase activity of each enzyme remained fully functional, as monitored retrospectively by release of free thymine after either photochemical-(5 kJ/m 2 , 254 nm) or photoenzymic- (Escherichia coli photolyase plus visible light) induced reversal of pyrimidine dimers in the UV-damaged substrate. The data demonstrate that the inhibition of the strand-incision reaction arises because of chemical modification of the AP sites and is not due to inactivation of the enzyme by methoxyamine. The results, combined with earlier findings for 5'-acting AP endonucleases, strongly suggest that methoxyamine is a highly specific inhibitor of virtually all AP endonucleases, irrespective of their modes of action, and may therefore prove useful in a wide variety of DNA repair studies

  14. Spectroelectrochemical insights into structural and redox properties of immobilized endonuclease III and its catalytically inactive mutant

    Science.gov (United States)

    Moe, Elin; Rollo, Filipe; Silveira, Célia M.; Sezer, Murat; Hildebrandt, Peter; Todorovic, Smilja

    2018-01-01

    Endonuclease III is a Fe-S containing bifunctional DNA glycosylase which is involved in the repair of oxidation damaged DNA. Here we employ surface enhanced IR spectroelectrochemistry and electrochemistry to study the enzyme from the highly radiation- and desiccation-resistant bacterium Deinococcus radiodurans (DrEndoIII2). The experiments are designed to shed more light onto specific parameters that are currently proposed to govern damage search and recognition by endonucleases III. We demonstrate that electrostatic interactions required for the redox activation of DrEndoIII2 may result in high electric fields that alter its structural and thermodynamic properties. Analysis of inactive DrEndoIII2 (K132A/D150A double mutant) interacting with undamaged DNA, and the active enzyme interacting with damaged DNA also indicate that the electron transfer is modulated by subtle differences in the protein-DNA complex.

  15. Dominant mutations in S. cerevisiae PMS1 identify the Mlh1-Pms1 endonuclease active site and an exonuclease 1-independent mismatch repair pathway.

    Science.gov (United States)

    Smith, Catherine E; Mendillo, Marc L; Bowen, Nikki; Hombauer, Hans; Campbell, Christopher S; Desai, Arshad; Putnam, Christopher D; Kolodner, Richard D

    2013-10-01

    Lynch syndrome (hereditary nonpolypsis colorectal cancer or HNPCC) is a common cancer predisposition syndrome. Predisposition to cancer in this syndrome results from increased accumulation of mutations due to defective mismatch repair (MMR) caused by a mutation in one of the mismatch repair genes MLH1, MSH2, MSH6 or PMS2/scPMS1. To better understand the function of Mlh1-Pms1 in MMR, we used Saccharomyces cerevisiae to identify six pms1 mutations (pms1-G683E, pms1-C817R, pms1-C848S, pms1-H850R, pms1-H703A and pms1-E707A) that were weakly dominant in wild-type cells, which surprisingly caused a strong MMR defect when present on low copy plasmids in an exo1Δ mutant. Molecular modeling showed these mutations caused amino acid substitutions in the metal coordination pocket of the Pms1 endonuclease active site and biochemical studies showed that they inactivated the endonuclease activity. This model of Mlh1-Pms1 suggested that the Mlh1-FERC motif contributes to the endonuclease active site. Consistent with this, the mlh1-E767stp mutation caused both MMR and endonuclease defects similar to those caused by the dominant pms1 mutations whereas mutations affecting the predicted metal coordinating residue Mlh1-C769 had no effect. These studies establish that the Mlh1-Pms1 endonuclease is required for MMR in a previously uncharacterized Exo1-independent MMR pathway.

  16. Dominant mutations in S. cerevisiae PMS1 identify the Mlh1-Pms1 endonuclease active site and an exonuclease 1-independent mismatch repair pathway.

    Directory of Open Access Journals (Sweden)

    Catherine E Smith

    2013-10-01

    Full Text Available Lynch syndrome (hereditary nonpolypsis colorectal cancer or HNPCC is a common cancer predisposition syndrome. Predisposition to cancer in this syndrome results from increased accumulation of mutations due to defective mismatch repair (MMR caused by a mutation in one of the mismatch repair genes MLH1, MSH2, MSH6 or PMS2/scPMS1. To better understand the function of Mlh1-Pms1 in MMR, we used Saccharomyces cerevisiae to identify six pms1 mutations (pms1-G683E, pms1-C817R, pms1-C848S, pms1-H850R, pms1-H703A and pms1-E707A that were weakly dominant in wild-type cells, which surprisingly caused a strong MMR defect when present on low copy plasmids in an exo1Δ mutant. Molecular modeling showed these mutations caused amino acid substitutions in the metal coordination pocket of the Pms1 endonuclease active site and biochemical studies showed that they inactivated the endonuclease activity. This model of Mlh1-Pms1 suggested that the Mlh1-FERC motif contributes to the endonuclease active site. Consistent with this, the mlh1-E767stp mutation caused both MMR and endonuclease defects similar to those caused by the dominant pms1 mutations whereas mutations affecting the predicted metal coordinating residue Mlh1-C769 had no effect. These studies establish that the Mlh1-Pms1 endonuclease is required for MMR in a previously uncharacterized Exo1-independent MMR pathway.

  17. Polymerase-endonuclease amplification reaction (PEAR for large-scale enzymatic production of antisense oligonucleotides.

    Directory of Open Access Journals (Sweden)

    Xiaolong Wang

    Full Text Available Antisense oligonucleotides targeting microRNAs or their mRNA targets prove to be powerful tools for molecular biology research and may eventually emerge as new therapeutic agents. Synthetic oligonucleotides are often contaminated with highly homologous failure sequences. Synthesis of a certain oligonucleotide is difficult to scale up because it requires expensive equipment, hazardous chemicals and a tedious purification process. Here we report a novel thermocyclic reaction, polymerase-endonuclease amplification reaction (PEAR, for the amplification of oligonucleotides. A target oligonucleotide and a tandem repeated antisense probe are subjected to repeated cycles of denaturing, annealing, elongation and cleaving, in which thermostable DNA polymerase elongation and strand slipping generate duplex tandem repeats, and thermostable endonuclease (PspGI cleavage releases monomeric duplex oligonucleotides. Each round of PEAR achieves over 100-fold amplification. The product can be used in one more round of PEAR directly, and the process can be further repeated. In addition to avoiding dangerous materials and improved product purity, this reaction is easy to scale up and amenable to full automation. PEAR has the potential to be a useful tool for large-scale production of antisense oligonucleotide drugs.

  18. RPA activates the XPF-ERCC1 endonuclease to initiate processing of DNA interstrand crosslinks.

    Science.gov (United States)

    Abdullah, Ummi B; McGouran, Joanna F; Brolih, Sanja; Ptchelkine, Denis; El-Sagheer, Afaf H; Brown, Tom; McHugh, Peter J

    2017-07-14

    During replication-coupled DNA interstrand crosslink (ICL) repair, the XPF-ERCC1 endonuclease is required for the incisions that release, or "unhook", ICLs, but the mechanism of ICL unhooking remains largely unknown. Incisions are triggered when the nascent leading strand of a replication fork strikes the ICL Here, we report that while purified XPF-ERCC1 incises simple ICL-containing model replication fork structures, the presence of a nascent leading strand, modelling the effects of replication arrest, inhibits this activity. Strikingly, the addition of the single-stranded DNA (ssDNA)-binding replication protein A (RPA) selectively restores XPF-ERCC1 endonuclease activity on this structure. The 5'-3' exonuclease SNM1A can load from the XPF-ERCC1-RPA-induced incisions and digest past the crosslink to quantitatively complete the unhooking reaction. We postulate that these collaborative activities of XPF-ERCC1, RPA and SNM1A might explain how ICL unhooking is achieved in vivo . © 2017 The Authors. Published under the terms of the CC BY 4.0 license.

  19. Restriction endonuclease analysis of Pasteurella multocida isolates from three California turkey premises.

    Science.gov (United States)

    Christiansen, K H; Carpenter, T E; Snipes, K P; Hird, D W; Ghazikhanian, G Y

    1992-01-01

    Three California turkey premises that had repeated outbreaks of fowl cholera were studied for periods of 2 to 4 years. Using biochemical, serologic, plasmid DNA, and restriction endonuclease analyses of isolates of Pasteurella multocida from turkeys and wildlife on the premises, strains of the organism were found to be enzootic on two of the premises. On the third, a variety of strains of P. multocida were isolated from fowl cholera outbreak flocks.

  20. Ultraviolet-endonuclease activity in cell extracts of Saccharomyces cerevisiae mutants defective in excision of pyrimidine dimers

    International Nuclear Information System (INIS)

    Bekker, M.L.; Kaboev, O.K.; Akhmedov, A.T.; Luchkina, L.A.

    1980-01-01

    Cell-free extracts of ultraviolet-sensitive mutants of Saccharomyces cerevisiae defective in excision of pyrimidine dimers, rad1, rad2, rad3, rad4, rad10, and rad16, as well as the extracts of the wild-type strain RAD+, display ultraviolet-endonuclease activity

  1. The structure-specific endonuclease Ercc1-Xpf is required for targeted gene replacement in embryonic stem cells

    NARCIS (Netherlands)

    L.J. Niedernhofer (Laura); J. Essers (Jeroen); G. Weeda (Geert); H.B. Beverloo (Berna); J. de Wit (Jan); M. Muijtjens (Manja); H. Odijk (Hanny); J.H.J. Hoeijmakers (Jan); R. Kanaar (Roland)

    2001-01-01

    textabstractThe Ercc1-Xpf heterodimer, a highly conserved structure-specific endonuclease, functions in multiple DNA repair pathways that are pivotal for maintaining genome stability, including nucleotide excision repair, interstrand crosslink repair and homologous recombination. Erccl-Xpf incises

  2. Geographically diverse Australian isolates of Melissococcus pluton exhibit minimal genotypic diversity by restriction endonuclease analysis.

    Science.gov (United States)

    Djordjevic, S P; Smith, L A; Forbes, W A; Hornitzky, M A

    1999-04-15

    Melissococcus pluton, the causative agent of European foulbrood is an economically significant disease of honey bees (Apis mellifera) across most regions of the world and is prevalent throughout most states of Australia. 49 Isolates of M. pluton recovered from diseased colonies or honey samples in New South Wales, Queensland, South Australia, Tasmania and Victoria were compared using SDS-PAGE, Western immunoblotting and restriction endonuclease analyses. DNA profiles of all 49 geographically diverse isolates showed remarkably similar AluI profiles although four isolates (one each from Queensland, South Australia, New South Wales and Victoria) displayed minor profile variations compared to AluI patterns of all other isolates. DNA from a subset of the 49 Australian and three isolates from the United Kingdom were digested separately with the restriction endonucleases CfoI, RsaI and DraI. Restriction endonuclease fragment patterns generated using these enzymes were also similar although minor variations were noted. SDS-PAGE of whole cell proteins from 13 of the 49 isolates from different states of Australia, including the four isolates which displayed minor profile variations (AluI) produced indistinguishable patterns. Major immunoreactive proteins of approximate molecular masses of 21, 24, 28, 30, 36, 40, 44, 56, 60, 71, 79 and 95 kDa were observed in immunoblots of whole cell lysates of 22 of the 49 isolates and reacted with rabbit hyperimmune antibodies raised against M. pluton whole cells. Neither SDS-PAGE or immunoblotting was capable of distinguishing differences between geographically diverse isolates of M. pluton. Collectively these data confirm that Australian isolates of M. pluton are genetically homogeneous and that this species may be clonal. Plasmid DNA was not detected in whole cell DNA profiles of any isolate resolved using agarose gel electrophoresis.

  3. Activation of Saccharomyces cerevisiae Mlh1-Pms1 Endonuclease in a Reconstituted Mismatch Repair System*

    Science.gov (United States)

    Smith, Catherine E.; Bowen, Nikki; Graham, William J.; Goellner, Eva M.; Srivatsan, Anjana; Kolodner, Richard D.

    2015-01-01

    Previous studies reported the reconstitution of an Mlh1-Pms1-independent 5′ nick-directed mismatch repair (MMR) reaction using Saccharomyces cerevisiae proteins. Here we describe the reconstitution of a mispair-dependent Mlh1-Pms1 endonuclease activation reaction requiring Msh2-Msh6 (or Msh2-Msh3), proliferating cell nuclear antigen (PCNA), and replication factor C (RFC) and a reconstituted Mlh1-Pms1-dependent 3′ nick-directed MMR reaction requiring Msh2-Msh6 (or Msh2-Msh3), exonuclease 1 (Exo1), replication protein A (RPA), RFC, PCNA, and DNA polymerase δ. Both reactions required Mg2+ and Mn2+ for optimal activity. The MMR reaction also required two reaction stages in which the first stage required incubation of Mlh1-Pms1 with substrate DNA, with or without Msh2-Msh6 (or Msh2-Msh3), PCNA, and RFC but did not require nicking of the substrate, followed by a second stage in which other proteins were added. Analysis of different mutant proteins demonstrated that both reactions required a functional Mlh1-Pms1 endonuclease active site, as well as mispair recognition and Mlh1-Pms1 recruitment by Msh2-Msh6 but not sliding clamp formation. Mutant Mlh1-Pms1 and PCNA proteins that were defective for Exo1-independent but not Exo1-dependent MMR in vivo were partially defective in the Mlh1-Pms1 endonuclease and MMR reactions, suggesting that both reactions reflect the activation of Mlh1-Pms1 seen in Exo1-independent MMR in vivo. The availability of this reconstituted MMR reaction should now make it possible to better study both Exo1-independent and Exo1-dependent MMR. PMID:26170454

  4. Activation of Saccharomyces cerevisiae Mlh1-Pms1 Endonuclease in a Reconstituted Mismatch Repair System.

    Science.gov (United States)

    Smith, Catherine E; Bowen, Nikki; Graham, William J; Goellner, Eva M; Srivatsan, Anjana; Kolodner, Richard D

    2015-08-28

    Previous studies reported the reconstitution of an Mlh1-Pms1-independent 5' nick-directed mismatch repair (MMR) reaction using Saccharomyces cerevisiae proteins. Here we describe the reconstitution of a mispair-dependent Mlh1-Pms1 endonuclease activation reaction requiring Msh2-Msh6 (or Msh2-Msh3), proliferating cell nuclear antigen (PCNA), and replication factor C (RFC) and a reconstituted Mlh1-Pms1-dependent 3' nick-directed MMR reaction requiring Msh2-Msh6 (or Msh2-Msh3), exonuclease 1 (Exo1), replication protein A (RPA), RFC, PCNA, and DNA polymerase δ. Both reactions required Mg(2+) and Mn(2+) for optimal activity. The MMR reaction also required two reaction stages in which the first stage required incubation of Mlh1-Pms1 with substrate DNA, with or without Msh2-Msh6 (or Msh2-Msh3), PCNA, and RFC but did not require nicking of the substrate, followed by a second stage in which other proteins were added. Analysis of different mutant proteins demonstrated that both reactions required a functional Mlh1-Pms1 endonuclease active site, as well as mispair recognition and Mlh1-Pms1 recruitment by Msh2-Msh6 but not sliding clamp formation. Mutant Mlh1-Pms1 and PCNA proteins that were defective for Exo1-independent but not Exo1-dependent MMR in vivo were partially defective in the Mlh1-Pms1 endonuclease and MMR reactions, suggesting that both reactions reflect the activation of Mlh1-Pms1 seen in Exo1-independent MMR in vivo. The availability of this reconstituted MMR reaction should now make it possible to better study both Exo1-independent and Exo1-dependent MMR. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  5. Purification and characterization of VDE, a site-specific endonuclease from the yeast Saccharomyces cerevisiae.

    Science.gov (United States)

    Gimble, F S; Thorner, J

    1993-10-15

    The 119-kDa primary translation product of the VMA1 gene of Saccharomyces cerevisiae undergoes a self-catalyzed rearrangement ("protein splicing") that excises an internal 50-kDa segment of the polypeptide and joins the amino-terminal and carboxyl-terminal segments to generate the 69-kDa subunit of the vacuolar membrane-associated H(+)-ATPase. We have shown previously that the internal segment is a site-specific endonuclease (Gimble, F. S., and Thorner, J. (1992) Nature 357, 301-306). Here we describe methods for the high level expression and purification to near homogeneity of both the authentic VMA1-derived endonuclease (or VDE) from yeast (yield 18%) and a recombinant form of VDE made in bacteria (yield 29%). Detailed characterization of these preparations demonstrated that the yeast-derived and bacterially produced enzymes were indistinguishable, as judged by: (a) behavior during purification; (b) apparent native molecular mass (50 kDa); (c) immunological reactivity; and (d) catalytic properties (specific activity; cleavage site recognition; and optima for pH, temperature, divalent cation and ionic strength). The minimal site required for VDE cleavage was delimited to a 30-base pair sequence within its specific substrate (the VMA1 delta vde allele).

  6. Biochemical characterization of recombinant influenza A polymerase heterotrimer complex: Endonuclease activity and evaluation of inhibitors

    Czech Academy of Sciences Publication Activity Database

    Xing, W.; Barauskas, O.; Kirschberg, T.; Niedziela-Majka, A.; Clarke, M.; Birkuš, Gabriel; Weissburg, P.; Liu, X.; Schultz, B. E.; Sakowicz, R.; Kwon, H. J.; Feng, J. Y.

    2017-01-01

    Roč. 12, č. 8 (2017), č. článku e0181969. E-ISSN 1932-6203 Institutional support: RVO:61388963 Keywords : virus PA endonuclease * respiratory syncytial virus * RNA synthesis Subject RIV: CE - Biochemistry OBOR OECD: Biochemistry and molecular biology Impact factor: 2.806, year: 2016 http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0181969

  7. An ultrasensitive colorimeter assay strategy for p53 mutation assisted by nicking endonuclease signal amplification.

    Science.gov (United States)

    Lin, Zhenyu; Yang, Weiqiang; Zhang, Guiyun; Liu, Qida; Qiu, Bin; Cai, Zongwei; Chen, Guonan

    2011-08-28

    A novel catalytic colorimetric assay assisted by nicking endonuclease signal amplification (NESA) was developed. With the signal amplification, the detection limit of the p53 target gene can be as low as 1 pM, which is nearly 5 orders of magnitude lower than that of other previously reported colorimetric DNA detection strategies based on catalytic DNAzyme.

  8. Identification and characterization of inhibitors of human apurinic/apyrimidinic endonuclease APE1.

    Directory of Open Access Journals (Sweden)

    Anton Simeonov

    2009-06-01

    Full Text Available APE1 is the major nuclease for excising abasic (AP sites and particular 3'-obstructive termini from DNA, and is an integral participant in the base excision repair (BER pathway. BER capacity plays a prominent role in dictating responsiveness to agents that generate oxidative or alkylation DNA damage, as well as certain chain-terminating nucleoside analogs and 5-fluorouracil. We describe within the development of a robust, 1536-well automated screening assay that employs a deoxyoligonucleotide substrate operating in the red-shifted fluorescence spectral region to identify APE1 endonuclease inhibitors. This AP site incision assay was used in a titration-based high-throughput screen of the Library of Pharmacologically Active Compounds (LOPAC(1280, a collection of well-characterized, drug-like molecules representing all major target classes. Prioritized hits were authenticated and characterized via two high-throughput screening assays -- a Thiazole Orange fluorophore-DNA displacement test and an E. coli endonuclease IV counterscreen -- and a conventional, gel-based radiotracer incision assay. The top, validated compounds, i.e. 6-hydroxy-DL-DOPA, Reactive Blue 2 and myricetin, were shown to inhibit AP site cleavage activity of whole cell protein extracts from HEK 293T and HeLa cell lines, and to enhance the cytotoxic and genotoxic potency of the alkylating agent methylmethane sulfonate. The studies herein report on the identification of novel, small molecule APE1-targeted bioactive inhibitor probes, which represent initial chemotypes towards the development of potential pharmaceuticals.

  9. Group I introns and associated homing endonuclease genes reveals a clinal structure for Porphyra spiralis var. amplifolia (Bangiales, Rhodophyta) along the Eastern coast of South America

    Science.gov (United States)

    2008-01-01

    Background Group I introns are found in the nuclear small subunit ribosomal RNA gene (SSU rDNA) of some species of the genus Porphyra (Bangiales, Rhodophyta). Size polymorphisms in group I introns has been interpreted as the result of the degeneration of homing endonuclease genes (HEG) inserted in peripheral loops of intron paired elements. In this study, intron size polymorphisms were characterized for different Porphyra spiralis var. amplifolia (PSA) populations on the Southern Brazilian coast, and were used to infer genetic relationships and genetic structure of these PSA populations, in addition to cox2-3 and rbcL-S regions. Introns of different sizes were tested qualitatively for in vitro self-splicing. Results Five intron size polymorphisms within 17 haplotypes were obtained from 80 individuals representing eight localities along the distribution of PSA in the Eastern coast of South America. In order to infer genetic structure and genetic relationships of PSA, these polymorphisms and haplotypes were used as markers for pairwise Fst analyses, Mantel's test and median joining network. The five cox2-3 haplotypes and the unique rbcL-S haplotype were used as markers for summary statistics, neutrality tests Tajima's D and Fu's Fs and for median joining network analyses. An event of demographic expansion from a population with low effective number, followed by a pattern of isolation by distance was obtained for PSA populations with the three analyses. In vitro experiments have shown that introns of different lengths were able to self-splice from pre-RNA transcripts. Conclusion The findings indicated that degenerated HEGs are reminiscent of the presence of a full-length and functional HEG, once fixed for PSA populations. The cline of HEG degeneration determined the pattern of isolation by distance. Analyses with the other markers indicated an event of demographic expansion from a population with low effective number. The different degrees of degeneration of the HEG

  10. Group I introns and associated homing endonuclease genes reveals a clinal structure for Porphyra spiralis var. amplifolia (Bangiales, Rhodophyta along the Eastern coast of South America

    Directory of Open Access Journals (Sweden)

    Matioli Sergio R

    2008-11-01

    Full Text Available Abstract Background Group I introns are found in the nuclear small subunit ribosomal RNA gene (SSU rDNA of some species of the genus Porphyra (Bangiales, Rhodophyta. Size polymorphisms in group I introns has been interpreted as the result of the degeneration of homing endonuclease genes (HEG inserted in peripheral loops of intron paired elements. In this study, intron size polymorphisms were characterized for different Porphyra spiralis var. amplifolia (PSA populations on the Southern Brazilian coast, and were used to infer genetic relationships and genetic structure of these PSA populations, in addition to cox2-3 and rbcL-S regions. Introns of different sizes were tested qualitatively for in vitro self-splicing. Results Five intron size polymorphisms within 17 haplotypes were obtained from 80 individuals representing eight localities along the distribution of PSA in the Eastern coast of South America. In order to infer genetic structure and genetic relationships of PSA, these polymorphisms and haplotypes were used as markers for pairwise Fst analyses, Mantel's test and median joining network. The five cox2-3 haplotypes and the unique rbcL-S haplotype were used as markers for summary statistics, neutrality tests Tajima's D and Fu's Fs and for median joining network analyses. An event of demographic expansion from a population with low effective number, followed by a pattern of isolation by distance was obtained for PSA populations with the three analyses. In vitro experiments have shown that introns of different lengths were able to self-splice from pre-RNA transcripts. Conclusion The findings indicated that degenerated HEGs are reminiscent of the presence of a full-length and functional HEG, once fixed for PSA populations. The cline of HEG degeneration determined the pattern of isolation by distance. Analyses with the other markers indicated an event of demographic expansion from a population with low effective number. The different degrees of

  11. Creation of targeted inversion mutations in plants using an RNA-guided endonuclease

    Institute of Scientific and Technical Information of China (English)

    Congsheng Zhang; Changlin Liu; Jianfeng Weng; Beijiu Cheng; Fang Liu; Xinhai Li; Chuanxiao Xie

    2017-01-01

    Inversions are DNA rearrangements that are essential for plant gene evolution and adaptation to environmental changes. We demonstrate the creation of targeted inversions and previously reported targeted deletion mutations via delivery of a pair of RNA-guided endonucleases (RGENs) of CRISPR/Cas9. The efficiencies of the targeted inversions were 2.6%and 2.2%in the Arabidopsis FLOWERING TIME (AtFT) and TERMINAL FLOWER 1 (AtTFL1) loci, respectively. Thus, we successfully established an approach that can potentially be used to introduce targeted DNA inversions of interest for functional studies and crop improvement.

  12. Alteration of Sequence Specificity of the Type IIS Restriction Endonuclease BtsI

    OpenAIRE

    Guan, Shengxi; Blanchard, Aine; Zhang, Penghua; Zhu, Zhenyu

    2010-01-01

    The Type IIS restriction endonuclease BtsI recognizes and digests at GCAGTG(2/0). It comprises two subunits: BtsIA and BtsIB. The BtsIB subunit contains the recognition domain, one catalytic domain for bottom strand nicking and part of the catalytic domain for the top strand nicking. BtsIA has the rest of the catalytic domain that is responsible for the DNA top strand nicking. BtsIA alone has no activity unless it mixes with BtsIB to reconstitute the BtsI activity. During characterization of ...

  13. A RecB-family nuclease motif in the Type I restriction endonuclease EcoR124I

    Czech Academy of Sciences Publication Activity Database

    Šišáková, Eva; Stanley, L. K.; Weiserová, Marie; Szczelkun, M. D.

    2008-01-01

    Roč. 36, č. 12 (2008), s. 1-11 ISSN 0305-1048 R&D Projects: GA ČR GA204/07/0325 Grant - others:XE(XE) BioNano-Switch 043288 Institutional research plan: CEZ:AV0Z50200510 Keywords : restriction endonuclease * mutagenesis * dsdna Subject RIV: EE - Microbiology, Virology Impact factor: 6.878, year: 2008

  14. Atypical myxomatosis--virus isolation, experimental infection of rabbits and restriction endonuclease analysis of the isolate.

    Science.gov (United States)

    Psikal, I; Smíd, B; Rodák, L; Valícek, L; Bendová, J

    2003-08-01

    Atypical form of myxomatosis, which caused non-lethal and clinically mild disease in domestic rabbits 1 month after immunization with a commercially available vaccine MXT, is described. The isolated myxoma virus designated as Litovel 2 (Li-2) did not induce systemic disease following subcutaneous and intradermal applications in susceptible experimental rabbits but led to the immune response demonstrated by ELISA. No severe disease was induced in those Li-2 inoculated rabbits by challenge with the virulent strains Lausanne (Lu) or Sanar (SA), while the control animals showed nodular form of myxomatosis with lethal course of the illness. Restriction fragment length polymorphism (RFLP) of genomic DNA with KpnI and BamHI endonucleases was used for genetic characterization of the Li-2 isolate, the vaccine strain MXT and both virulent strains Lu and SA, respectively. In general, RFLP analysis has shown to be informative for inferring genetic relatedness between myxoma viruses. Based on restriction endonuclease DNA fragment size distribution, it was evident that the pathogenic strain SA is genetically related to the reference strain Lu and the isolate Li-2 is more related, but not identical, to the vaccination strain MXT.

  15. Single substitution in bacteriophage T4 RNase H alters the ratio between its exo- and endonuclease activities.

    Science.gov (United States)

    Kholod, Natalia; Sivogrivov, Dmitry; Latypov, Oleg; Mayorov, Sergey; Kuznitsyn, Rafail; Kajava, Andrey V; Shlyapnikov, Mikhail; Granovsky, Igor

    2015-11-01

    The article describes substitutions in bacteriophage T4 RNase H which provide so called das-effect. Phage T4 DNA arrest suppression (das) mutations have been described to be capable of partially suppressing the phage DNA arrest phenotype caused by a dysfunction in genes 46 and/or 47 (also known as Mre11/Rad50 complex). Genetic mapping of das13 (one of the das mutations) has shown it to be in the region of the rnh gene encoding RNase H. Here we report that Das13 mutant of RNase H has substitutions of valine 43 and leucine 242 with isoleucines. To investigate the influence of these mutations on RNase H nuclease properties we have designed a novel in vitro assay that allows us to separate and quantify exo- or endonuclease activities of flap endonuclease. The nuclease assay in vitro showed that V43I substitution increased the ratio between exonuclease/endonuclease activities of RNase H whereas L242I substitution did not affect the nuclease activity of RNase H in vitro. However, both mutations were necessary for the full das effect in vivo. Molecular modelling of the nuclease structure suggests that V43I substitution may lead to disposition of H4 helix, responsible for the interaction with the first base pairs of 5'end of branched DNA. These structural changes may affect unwinding of the first base pairs of gapped or nicked DNA generating a short flap and therefore may stabilize the DNA-enzyme complex. L242I substitution did not affect the structure of RNase H and its role in providing das-effect remains unclear. Copyright © 2015 Elsevier B.V. All rights reserved.

  16. Selective metal binding to Cys-78 within endonuclease V causes an inhibition of catalytic activities without altering nontarget and target DNA binding

    International Nuclear Information System (INIS)

    Prince, M.A.; Friedman, B.; Gruskin, E.A.; Schrock, R.D. III; Lloyd, R.S.

    1991-01-01

    T4 endonuclease V is a pyrimidine dimer-specific DNA repair enzyme which has been previously shown not to require metal ions for either of its two catalytic activities or its DNA binding function. However, we have investigated whether the single cysteine within the enzyme was able to bind metal salts and influence the various activities of this repair enzyme. A series of metals (Hg2+, Ag+, Cu+) were shown to inactivate both endonuclease Vs pyrimidine dimer-specific DNA glycosylase activity and the subsequent apurinic nicking activity. The binding of metal to endonuclease V did not interfere with nontarget DNA scanning or pyrimidine dimer-specific binding. The Cys-78 codon within the endonuclease V gene was changed by oligonucleotide site-directed mutagenesis to Thr-78 and Ser-78 in order to determine whether the native cysteine was directly involved in the enzyme's DNA catalytic activities and whether the cysteine was primarily responsible for the metal binding. The mutant enzymes were able to confer enhanced ultraviolet light (UV) resistance to DNA repair-deficient Escherichia coli at levels equal to that conferred by the wild type enzyme. The C78T mutant enzyme was purified to homogeneity and shown to be catalytically active on pyrimidine dimer-containing DNA. The catalytic activities of the C78T mutant enzyme were demonstrated to be unaffected by the addition of Hg2+ or Ag+ at concentrations 1000-fold greater than that required to inhibit the wild type enzyme. These data suggest that the cysteine is not required for enzyme activity but that the binding of certain metals to that amino acid block DNA incision by either preventing a conformational change in the enzyme after it has bound to a pyrimidine dimer or sterically interfering with the active site residue's accessibility to the pyrimidine dimer

  17. Apurinic/apyrimidinic endonucleases of Mycobacterium tuberculosis protect against DNA damage but are dispensable for the growth of the pathogen in guinea pigs.

    Science.gov (United States)

    Puri, Rupangi Verma; Reddy, P Vineel; Tyagi, Anil K

    2014-01-01

    In host cells, Mycobacterium tuberculosis encounters an array of reactive molecules capable of damaging its genome. Non-bulky DNA lesions are the most common damages produced on the exposure of the pathogen to reactive species and base excision repair (BER) pathway is involved in the repair of such damage. During BER, apurinic/apyrimidinic (AP) endonuclease enzymes repair the abasic sites that are generated after spontaneous DNA base loss or by the action of DNA glycosylases, which if left unrepaired lead to inhibition of replication and transcription. However, the role of AP endonucleases in imparting protection against DNA damage and in the growth and pathogenesis of M.tuberculosis has not yet been elucidated. To demonstrate the biological significance of these enzymes in M.tuberculosis, it would be desirable to disrupt the relevant genes and evaluate the resulting mutants for their ability to grow in the host and cause disease. In this study, we have generated M.tuberculosis mutants of the base excision repair (BER) system, disrupted in either one (MtbΔend or MtbΔxthA) or both the AP endonucleases (MtbΔendΔxthA). We demonstrate that these genes are crucial for bacteria to withstand alkylation and oxidative stress in vitro. In addition, the mutant disrupted in both the AP endonucleases (MtbΔendΔxthA) exhibited a significant reduction in its ability to survive inside human macrophages. However, infection of guinea pigs with either MtbΔend or MtbΔxthA or MtbΔendΔxthA resulted in the similar bacillary load and pathological damage in the organs as observed in the case of infection with wild-type M.tuberculosis. The implications of these observations are discussed.

  18. Apurinic/apyrimidinic endonucleases of Mycobacterium tuberculosis protect against DNA damage but are dispensable for the growth of the pathogen in guinea pigs.

    Directory of Open Access Journals (Sweden)

    Rupangi Verma Puri

    Full Text Available In host cells, Mycobacterium tuberculosis encounters an array of reactive molecules capable of damaging its genome. Non-bulky DNA lesions are the most common damages produced on the exposure of the pathogen to reactive species and base excision repair (BER pathway is involved in the repair of such damage. During BER, apurinic/apyrimidinic (AP endonuclease enzymes repair the abasic sites that are generated after spontaneous DNA base loss or by the action of DNA glycosylases, which if left unrepaired lead to inhibition of replication and transcription. However, the role of AP endonucleases in imparting protection against DNA damage and in the growth and pathogenesis of M.tuberculosis has not yet been elucidated. To demonstrate the biological significance of these enzymes in M.tuberculosis, it would be desirable to disrupt the relevant genes and evaluate the resulting mutants for their ability to grow in the host and cause disease. In this study, we have generated M.tuberculosis mutants of the base excision repair (BER system, disrupted in either one (MtbΔend or MtbΔxthA or both the AP endonucleases (MtbΔendΔxthA. We demonstrate that these genes are crucial for bacteria to withstand alkylation and oxidative stress in vitro. In addition, the mutant disrupted in both the AP endonucleases (MtbΔendΔxthA exhibited a significant reduction in its ability to survive inside human macrophages. However, infection of guinea pigs with either MtbΔend or MtbΔxthA or MtbΔendΔxthA resulted in the similar bacillary load and pathological damage in the organs as observed in the case of infection with wild-type M.tuberculosis. The implications of these observations are discussed.

  19. A newly discovered Bordetella species carries a transcriptionally active CRISPR-Cas with a small Cas9 endonuclease

    Science.gov (United States)

    The Cas9 endonuclease of the Type II-a clustered regularly interspersed short palindromic repeats (CRISPR), of Streptococcus pyogenes (SpCas9) has been adapted as a widely used tool for genome editing and genome engineering. Herein, we describe a gene encoding a novel Cas9 ortholog (BpsuCas9) and th...

  20. Analysis of Endonuclease R·EcoRI Fragments of DNA from Lambdoid Bacteriophages and Other Viruses by Agarose-Gel Electrophoresis

    Science.gov (United States)

    Helling, Robert B.; Goodman, Howard M.; Boyer, Herbert W.

    1974-01-01

    By means of agarose-gel electrophoresis, endonuclease R·EcoRI-generated fragments of DNA from various viruses were separated, their molecular weights were determined, and complete or partial fragment maps for lambda, φ80, and hybrid phages were constructed. Images PMID:4372397

  1. Multiplex, rapid and sensitive isothermal detection of nucleic-acid sequence by endonuclease restriction-mediated real-time multiple cross displacement amplification

    Directory of Open Access Journals (Sweden)

    Yi eWang

    2016-05-01

    Full Text Available We have devised a novel isothermal amplification technology, termed endonuclease restriction-mediated real-time multiple cross displacement amplification (ET-MCDA, which facilitated multiplex, rapid, specific and sensitive detection of nucleic-acid sequences at a constant temperature. The ET-MCDA integrated multiple cross displacement amplification strategy, restriction endonuclease cleavage and real-time fluorescence detection technique. In the ET-MCDA system, the functional cross primer E-CP1 or E-CP2 was constructed by adding a short sequence at the 5’ end of CP1 or CP2, respectively, and the new E-CP1 or E-CP2 primer was labelled at the 5’ end with a fluorophore and in the middle with a dark quencher. The restriction endonuclease Nb.BsrDI specifically recognized the short sequence and digested the newly synthesized double-stranded terminal sequences (5’ end short sequences and their complementary sequences, which released the quenching, resulting on a gain of fluorescence signal. Thus, the ET-MCDA allowed real-time detection of single or multiple targets in only a single reaction, and the positive results were observed in as short as 12 minutes, detecting down to 3.125 fg of genomic DNA per tube. Moreover, the analytical specificity and the practical application of the ET-MCDA were also successfully evaluated in this study. Here we provided the details on the novel ET-MCDA technique and expounded the basic ET-MCDA amplification mechanism.

  2. Multiplex, Rapid, and Sensitive Isothermal Detection of Nucleic-Acid Sequence by Endonuclease Restriction-Mediated Real-Time Multiple Cross Displacement Amplification.

    Science.gov (United States)

    Wang, Yi; Wang, Yan; Zhang, Lu; Liu, Dongxin; Luo, Lijuan; Li, Hua; Cao, Xiaolong; Liu, Kai; Xu, Jianguo; Ye, Changyun

    2016-01-01

    We have devised a novel isothermal amplification technology, termed endonuclease restriction-mediated real-time multiple cross displacement amplification (ET-MCDA), which facilitated multiplex, rapid, specific and sensitive detection of nucleic-acid sequences at a constant temperature. The ET-MCDA integrated multiple cross displacement amplification strategy, restriction endonuclease cleavage and real-time fluorescence detection technique. In the ET-MCDA system, the functional cross primer E-CP1 or E-CP2 was constructed by adding a short sequence at the 5' end of CP1 or CP2, respectively, and the new E-CP1 or E-CP2 primer was labeled at the 5' end with a fluorophore and in the middle with a dark quencher. The restriction endonuclease Nb.BsrDI specifically recognized the short sequence and digested the newly synthesized double-stranded terminal sequences (5' end short sequences and their complementary sequences), which released the quenching, resulting on a gain of fluorescence signal. Thus, the ET-MCDA allowed real-time detection of single or multiple targets in only a single reaction, and the positive results were observed in as short as 12 min, detecting down to 3.125 fg of genomic DNA per tube. Moreover, the analytical specificity and the practical application of the ET-MCDA were also successfully evaluated in this study. Here, we provided the details on the novel ET-MCDA technique and expounded the basic ET-MCDA amplification mechanism.

  3. Cleavage of phosphorothioated DNA and methylated DNA by the type IV restriction endonuclease ScoMcrA.

    Directory of Open Access Journals (Sweden)

    Guang Liu

    2010-12-01

    Full Text Available Many taxonomically diverse prokaryotes enzymatically modify their DNA by replacing a non-bridging oxygen with a sulfur atom at specific sequences. The biological implications of this DNA S-modification (phosphorothioation were unknown. We observed that simultaneous expression of the dndA-E gene cluster from Streptomyces lividans 66, which is responsible for the DNA S-modification, and the putative Streptomyces coelicolor A(32 Type IV methyl-dependent restriction endonuclease ScoA3McrA (Sco4631 leads to cell death in the same host. A His-tagged derivative of ScoA3McrA cleaved S-modified DNA and also Dcm-methylated DNA in vitro near the respective modification sites. Double-strand cleavage occurred 16-28 nucleotides away from the phosphorothioate links. DNase I footprinting demonstrated binding of ScoA3McrA to the Dcm methylation site, but no clear binding could be detected at the S-modified site under cleavage conditions. This is the first report of in vitro endonuclease activity of a McrA homologue and also the first demonstration of an enzyme that specifically cleaves S-modified DNA.

  4. Microinjection of Micrococcus luteus UV-endonuclease restores UV-induced unscheduled DNA synthesis in cells of 9 xeroderma pigmentosum complementation groups.

    NARCIS (Netherlands)

    A.J.R. de Jonge; W. Vermeulen (Wim); W. Keijzer; J.H.J. Hoeijmakers (Jan); D. Bootsma (Dirk)

    1985-01-01

    textabstractThe UV-induced unscheduled DNA synthesis (UDS) in cultured cells of excision-deficient xeroderma pigmentosum (XP) complementation groups A through I was assayed after injection of Micrococcus luteus UV-endonuclease using glass microneedles. In all complementation groups a restoration of

  5. Identification of flap structure-specific endonuclease 1 as a factor involved in long-term memory formation of aversive learning.

    Science.gov (United States)

    Saavedra-Rodríguez, Lorena; Vázquez, Adrinel; Ortiz-Zuazaga, Humberto G; Chorna, Nataliya E; González, Fernando A; Andrés, Lissette; Rodríguez, Karen; Ramírez, Fernando; Rodríguez, Alan; Peña de Ortiz, Sandra

    2009-05-06

    We previously proposed that DNA recombination/repair processes play a role in memory formation. Here, we examined the possible role of the fen-1 gene, encoding a flap structure-specific endonuclease, in memory consolidation of conditioned taste aversion (CTA). Quantitative real-time PCR showed that amygdalar fen-1 mRNA induction was associated to the central processing of the illness experience related to CTA and to CTA itself, but not to the central processing resulting from the presentation of a novel flavor. CTA also increased expression of the Fen-1 protein in the amygdala, but not the insular cortex. In addition, double immunofluorescence analyses showed that amygdalar Fen-1 expression is mostly localized within neurons. Importantly, functional studies demonstrated that amygdalar antisense knockdown of fen-1 expression impaired consolidation, but not short-term memory, of CTA. Overall, these studies define the fen-1 endonuclease as a new DNA recombination/repair factor involved in the formation of long-term memories.

  6. VDE-initiated intein homing in Saccharomyces cerevisiae proceeds in a meiotic recombination-like manner.

    Science.gov (United States)

    Fukuda, Tomoyuki; Nogami, Satoru; Ohya, Yoshikazu

    2003-07-01

    Inteins and group I introns found in prokaryotic and eukaryotic organisms occasionally behave as mobile genetic elements. During meiosis of the yeast Saccharomyces cerevisiae, the site-specific endonuclease encoded by VMA1 intein, VDE, triggers a single double-strand break (DSB) at an inteinless allele, leading to VMA1 intein homing. Besides the accumulating information on the in vitro activity of VDE, very little has been known about the molecular mechanism of intein homing in yeast nucleus. We developed an assay to detect the product of VMA1 intein homing in yeast genome. We analysed mutant phenotypes of RecA homologs, Rad51p and Dmc1p, and their interacting proteins, Rad54p and Tid1p, and found that they all play critical roles in intein inheritance. The absence of DSB end processing proteins, Sae2p and those in the Mre11-Rad50-Xrs2 complex, also causes partial reduction in homing efficiency. As with meiotic recombination, crossover events are frequently observed during intein homing. We also observed that the absence of premeiotic DNA replication caused by hydroxyurea (HU) or clb5delta clb6delta mutation reduces VDE-mediated DSBs. The repairing system working in intein homing shares molecular machinery with meiotic recombination induced by Spo11p. Moreover, like Spo11p-induced DNA cleavage, premeiotic DNA replication is a prerequisite for a VDE-induced DSB. VMA1 intein thus utilizes several host factors involved in meiotic and recombinational processes to spread its genetic information and guarantee its progeny through establishment of a parasitic relationship with the organism.

  7. Spectrophotometric, colorimetric and visually detection of Pseudomonas aeruginosa ETA gene based gold nanoparticles DNA probe and endonuclease enzyme

    Science.gov (United States)

    Amini, Bahram; Kamali, Mehdi; Salouti, Mojtaba; Yaghmaei, Parichehreh

    2018-06-01

    Colorimetric DNA detection is preferred over other methods for clinical molecular diagnosis because it does not require expensive equipment. In the present study, the colorimetric method based on gold nanoparticles (GNPs) and endonuclease enzyme was used for the detection of P. aeruginosa ETA gene. Firstly, the primers and probe for P. aeruginosa exotoxin A (ETA) gene were designed and checked for specificity by the PCR method. Then, GNPs were synthesized using the citrate reduction method and conjugated with the prepared probe to develop the new nano-biosensor. Next, the extracted target DNA of the bacteria was added to GNP-probe complex to check its efficacy for P. aeruginosa ETA gene diagnosis. A decrease in absorbance was seen when GNP-probe-target DNA cleaved into the small fragments of BamHI endonuclease due to the weakened electrostatic interaction between GNPs and the shortened DNA. The right shift of the absorbance peak from 530 to 562 nm occurred after adding the endonuclease. It was measured using a UV-VIS absorption spectroscopy that indicates the existence of the P. aeruginosa ETA gene. Sensitivity was determined in the presence of different concentrations of target DNA of P. aeruginosa. The results obtained from the optimized conditions showed that the absorbance value has linear correlation with concentration of target DNA (R: 0.9850) in the range of 10-50 ng mL-1 with the limit detection of 9.899 ng mL-1. Thus, the specificity of the new method for detection of P. aeruginosa was established in comparison with other bacteria. Additionally, the designed assay was quantitatively applied to detect the P. aeruginosa ETA gene from 103 to 108 CFU mL-1 in real samples with a detection limit of 320 CFU mL-1.

  8. Molecular mechanisms involved in the production of chromosomal aberrations. I. Utilization of Neurospora endonuclease for the study of aberration production in G2 stage of the cell cycle

    Energy Technology Data Exchange (ETDEWEB)

    Natarajan, A T; Obe, G [Rijksuniversiteit Leiden (Netherlands). J.A. Cohen Inst. voor Radiopathologie en Stralingsbescherming

    1978-10-01

    Chinese hamster ovary cells (CHO) were X-irradiated in G2 stage of the cell cycle and immediately treated, in the presence of inactivated Sendai virus, with Neurospora endonuclease (E.C. 3.1.4.), an enzyme which is specific for cleaving single-stranded DNA. With this treatment, the frequencies of all types of chromosome aberrations increased when compared to X-irradiated controls. These results are interpreted as due to the conversion of some of the X-ray induced single-stranded DNA breaks into double-strand breaks by this enzyme. Similar enhancement due to this enzyme was found following treatment with methyl methanesulfonate (MMS) and bleomycin, but not following UV and mitomycin C. Addition of Micrococcus endonuclease and Neurospora endonuclease to the cells did not alter the frequencies of aberrations induced by UV. The introduction of enzymes with specific DNA-repair function offers possibilities to probe into the molecular events involved in the formation of structural chromosome aberrations induced by different classes of physical and chemical mutagens.

  9. Specificity of binding to four-way junctions in DNA by bacteriophage T7 endonuclease I.

    OpenAIRE

    Parsons, C A; West, S C

    1990-01-01

    T7 endonuclease I binds specifically to four-way junctions in duplex DNA and promotes their resolution into linear duplexes. Under conditions in which the nuclease activity is blocked by the absence of divalent cations, the enzyme forms a distinct protein-DNA complex with the junction, as detected by gel retardation and filter binding assays. The formation of this complex is structure-specific and contrasts with the short-lived binding complexes formed on linear duplex DNA. The binding comple...

  10. An ultra-sensitive colorimetric Hg(2+)-sensing assay based on DNAzyme-modified Au NP aggregation, MNPs and an endonuclease.

    Science.gov (United States)

    Li, Chao; Dai, Peiqing; Rao, Xinyi; Shao, Lin; Cheng, Guifang; He, Pingang; Fang, Yuzhi

    2015-01-01

    This paper reports the development of an ultra-sensitive colorimetric method for the detection of trace mercury ions involving DNAzymes, Au nanoparticle aggregation, magnetic nanoparticles and an endonuclease. DNAzyme-sensing elements are conjugated to the surface of Au nanoparticle-2, which can crosslink with the T-rich strands coated on Au nanoparticle-1 to form Au nanoparticle aggregation. Other T-rich stands are immobilized on the surface of MNPs. The specific hybridization of these two T-rich strands depends on the presence of Hg(2+), resulting in the formation of a T-Hg(2+)-T structure. Added endonuclease then digests the hybridized strands, and DNAzyme-modified Au NP aggregation is released, catalysing the conversion of the colourless ABTS into a blue-green product by H2O2-mediated oxidation. The increase in the adsorption spectrum of ABTS(+) at 421 nm is related to the concentration of Hg(2+). This assay was validated by detecting mercury ion concentrations in river water. The colorimetric responses were not significantly altered in the presence of 100-fold excesses of other metal ions such as Zn(2+), Pb(2+), Cd(2+), Mn(2+), Ca(2+) and Ni(2+). The inclusion of both Au NP aggregation and an endonuclease enables the assay to eliminate interference from the magnetic nanoparticles with colorimetric detection, decrease the background and improve the detection sensitivity. The calibration curve of the assay was linear over the range of Hg(2+) concentrations from 1 to 30 nM, and the detection limit was 0.8 nM, which is far lower than the 10 nM US EPA limit for drinking water. Copyright © 2014 Elsevier B.V. All rights reserved.

  11. Restoration of ultraviolet-induced unscheduled DNA synthesis of xeroderma pigmentosum cells by the concomitant treatment with bacteriophage T4 endonuclease V and HVJ (Sendai virus)

    International Nuclear Information System (INIS)

    Tanaka, K.; Sekiguchi, M.; Okada, Y.

    1975-01-01

    Ultraviolet (uv)-induced unscheduled DNA synthesis of xeroderma pigmentosum cells, belonging to complementation groups, A, B, C, D, and E, was restored to the normal level by concomitant treatment of the cells with T4 endonuclease V and uv-inactivated HVJ (Sendai virus). The present results suggest that T4 endonuclease molecules were inserted effectively into the cells by the interaction of HVJ with the cell membranes, the enzyme was functional on human chromosomal DNA which had been damaged by uv irradiation in the viable cells, all the studied groups of xeroderma pigmentosum (variant was not tested) were defective in the first step (incision) of excision repair

  12. A Mismatch EndoNuclease Array-Based Methodology (MENA) for Identifying Known SNPs or Novel Point Mutations.

    Science.gov (United States)

    Comeron, Josep M; Reed, Jordan; Christie, Matthew; Jacobs, Julia S; Dierdorff, Jason; Eberl, Daniel F; Manak, J Robert

    2016-04-05

    Accurate and rapid identification or confirmation of single nucleotide polymorphisms (SNPs), point mutations and other human genomic variation facilitates understanding the genetic basis of disease. We have developed a new methodology (called MENA (Mismatch EndoNuclease Array)) pairing DNA mismatch endonuclease enzymology with tiling microarray hybridization in order to genotype both known point mutations (such as SNPs) as well as identify previously undiscovered point mutations and small indels. We show that our assay can rapidly genotype known SNPs in a human genomic DNA sample with 99% accuracy, in addition to identifying novel point mutations and small indels with a false discovery rate as low as 10%. Our technology provides a platform for a variety of applications, including: (1) genotyping known SNPs as well as confirming newly discovered SNPs from whole genome sequencing analyses; (2) identifying novel point mutations and indels in any genomic region from any organism for which genome sequence information is available; and (3) screening panels of genes associated with particular diseases and disorders in patient samples to identify causative mutations. As a proof of principle for using MENA to discover novel mutations, we report identification of a novel allele of the beethoven (btv) gene in Drosophila, which encodes a ciliary cytoplasmic dynein motor protein important for auditory mechanosensation.

  13. Restriction endonucleases from invasive Neisseria gonorrhoeae cause double-strand breaks and distort mitosis in epithelial cells during infection.

    Directory of Open Access Journals (Sweden)

    Linda Weyler

    Full Text Available The host epithelium is both a barrier against, and the target for microbial infections. Maintaining regulated cell growth ensures an intact protective layer towards microbial-induced cellular damage. Neisseria gonorrhoeae infections disrupt host cell cycle regulation machinery and the infection causes DNA double strand breaks that delay progression through the G2/M phase. We show that intracellular gonococci upregulate and release restriction endonucleases that enter the nucleus and damage human chromosomal DNA. Bacterial lysates containing restriction endonucleases were able to fragment genomic DNA as detected by PFGE. Lysates were also microinjected into the cytoplasm of cells in interphase and after 20 h, DNA double strand breaks were identified by 53BP1 staining. In addition, by using live-cell microscopy and NHS-ester stained live gonococci we visualized the subcellular location of the bacteria upon mitosis. Infected cells show dysregulation of the spindle assembly checkpoint proteins MAD1 and MAD2, impaired and prolonged M-phase, nuclear swelling, micronuclei formation and chromosomal instability. These data highlight basic molecular functions of how gonococcal infections affect host cell cycle regulation, cause DNA double strand breaks and predispose cellular malignancies.

  14. Restriction endonucleases from invasive Neisseria gonorrhoeae cause double-strand breaks and distort mitosis in epithelial cells during infection.

    Science.gov (United States)

    Weyler, Linda; Engelbrecht, Mattias; Mata Forsberg, Manuel; Brehwens, Karl; Vare, Daniel; Vielfort, Katarina; Wojcik, Andrzej; Aro, Helena

    2014-01-01

    The host epithelium is both a barrier against, and the target for microbial infections. Maintaining regulated cell growth ensures an intact protective layer towards microbial-induced cellular damage. Neisseria gonorrhoeae infections disrupt host cell cycle regulation machinery and the infection causes DNA double strand breaks that delay progression through the G2/M phase. We show that intracellular gonococci upregulate and release restriction endonucleases that enter the nucleus and damage human chromosomal DNA. Bacterial lysates containing restriction endonucleases were able to fragment genomic DNA as detected by PFGE. Lysates were also microinjected into the cytoplasm of cells in interphase and after 20 h, DNA double strand breaks were identified by 53BP1 staining. In addition, by using live-cell microscopy and NHS-ester stained live gonococci we visualized the subcellular location of the bacteria upon mitosis. Infected cells show dysregulation of the spindle assembly checkpoint proteins MAD1 and MAD2, impaired and prolonged M-phase, nuclear swelling, micronuclei formation and chromosomal instability. These data highlight basic molecular functions of how gonococcal infections affect host cell cycle regulation, cause DNA double strand breaks and predispose cellular malignancies.

  15. Thermographic measurements of the spin Peltier effect in metal/yttrium-iron-garnet junction systems

    Science.gov (United States)

    Daimon, Shunsuke; Uchida, Ken-ichi; Iguchi, Ryo; Hioki, Tomosato; Saitoh, Eiji

    2017-07-01

    The spin Peltier effect (SPE), heat-current generation due to spin-current injection, in various metal (Pt, W, and Au single layers and Pt/Cu bilayer)/ferrimagnetic insulator [yttrium-iron-garnet (YIG)] junction systems has been investigated by means of a lock-in thermography (LIT) method. The SPE is excited by a spin current across the metal/YIG interface, which is generated by applying a charge current to the metallic layer via the spin Hall effect. The LIT method enables the thermal imaging of the SPE free from the Joule-heating contribution. Importantly, we observed spin-current-induced temperature modulation not only in the Pt/YIG and W/YIG systems, but also in the Au/YIG and Pt/Cu/YIG systems, excluding the possible contamination by anomalous Ettingshausen effects due to proximity-induced ferromagnetism near the metal/YIG interface. As demonstrated in our previous study, the SPE signals are confined only in the vicinity of the metal/YIG interface; we buttress this conclusion by reducing a spatial blur due to thermal diffusion in an infrared-emission layer on the sample surface used for the LIT measurements. We also found that the YIG-thickness dependence of the SPE is similar to that of the spin Seebeck effect measured in the same Pt/YIG sample, implying the reciprocal relation between them.

  16. Apoptotic DNA Degradation into Oligonucleosomal Fragments, but Not Apoptotic Nuclear Morphology, Relies on a Cytosolic Pool of DFF40/CAD Endonuclease*

    Science.gov (United States)

    Iglesias-Guimarais, Victoria; Gil-Guiñon, Estel; Gabernet, Gisela; García-Belinchón, Mercè; Sánchez-Osuna, María; Casanelles, Elisenda; Comella, Joan X.; Yuste, Victor J.

    2012-01-01

    Apoptotic cell death is characterized by nuclear fragmentation and oligonucleosomal DNA degradation, mediated by the caspase-dependent specific activation of DFF40/CAD endonuclease. Here, we describe how, upon apoptotic stimuli, SK-N-AS human neuroblastoma-derived cells show apoptotic nuclear morphology without displaying concomitant internucleosomal DNA fragmentation. Cytotoxicity afforded after staurosporine treatment is comparable with that obtained in SH-SY5Y cells, which exhibit a complete apoptotic phenotype. SK-N-AS cell death is a caspase-dependent process that can be impaired by the pan-caspase inhibitor q-VD-OPh. The endogenous inhibitor of DFF40/CAD, ICAD, is correctly processed, and dff40/cad cDNA sequence does not reveal mutations altering its amino acid composition. Biochemical approaches show that both SH-SY5Y and SK-N-AS resting cells express comparable levels of DFF40/CAD. However, the endonuclease is poorly expressed in the cytosolic fraction of healthy SK-N-AS cells. Despite this differential subcellular distribution of DFF40/CAD, we find no differences in the subcellular localization of both pro-caspase-3 and ICAD between the analyzed cell lines. After staurosporine treatment, the preferential processing of ICAD in the cytosolic fraction allows the translocation of DFF40/CAD from this fraction to a chromatin-enriched one. Therefore, the low levels of cytosolic DFF40/CAD detected in SK-N-AS cells determine the absence of DNA laddering after staurosporine treatment. In these cells DFF40/CAD cytosolic levels can be restored by the overexpression of their own endonuclease, which is sufficient to make them proficient at degrading their chromatin into oligonucleosome-size fragments after staurosporine treatment. Altogether, the cytosolic levels of DFF40/CAD are determinants in achieving a complete apoptotic phenotype, including oligonucleosomal DNA degradation. PMID:22253444

  17. Quantitative Temperature Dependence of Longitudinal Spin Seebeck Effect at High Temperatures

    Directory of Open Access Journals (Sweden)

    Ken-ichi Uchida

    2014-11-01

    Full Text Available We report temperature-dependent measurements of longitudinal spin Seebeck effects (LSSEs in Pt/Y_{3}Fe_{5}O_{12} (YIG/Pt systems in a high temperature range from room temperature to above the Curie temperature of YIG. The experimental results show that the magnitude of the LSSE voltage in the Pt/YIG/Pt systems rapidly decreases with increasing the temperature and disappears above the Curie temperature. The critical exponent of the LSSE voltage in the Pt/YIG/Pt systems at the Curie temperature is estimated to be 3, which is much greater than that for the magnetization curve of YIG. This difference highlights the fact that the mechanism of the LSSE cannot be explained in terms of simple static magnetic properties in YIG.

  18. DNA interrogation by the CRISPR RNA-guided endonuclease Cas9.

    Science.gov (United States)

    Sternberg, Samuel H; Redding, Sy; Jinek, Martin; Greene, Eric C; Doudna, Jennifer A

    2014-03-06

    The clustered regularly interspaced short palindromic repeats (CRISPR)-associated enzyme Cas9 is an RNA-guided endonuclease that uses RNA-DNA base-pairing to target foreign DNA in bacteria. Cas9-guide RNA complexes are also effective genome engineering agents in animals and plants. Here we use single-molecule and bulk biochemical experiments to determine how Cas9-RNA interrogates DNA to find specific cleavage sites. We show that both binding and cleavage of DNA by Cas9-RNA require recognition of a short trinucleotide protospacer adjacent motif (PAM). Non-target DNA binding affinity scales with PAM density, and sequences fully complementary to the guide RNA but lacking a nearby PAM are ignored by Cas9-RNA. Competition assays provide evidence that DNA strand separation and RNA-DNA heteroduplex formation initiate at the PAM and proceed directionally towards the distal end of the target sequence. Furthermore, PAM interactions trigger Cas9 catalytic activity. These results reveal how Cas9 uses PAM recognition to quickly identify potential target sites while scanning large DNA molecules, and to regulate scission of double-stranded DNA.

  19. DNA interrogation by the CRISPR RNA-guided endonuclease Cas9

    Science.gov (United States)

    Sternberg, Samuel H.; Redding, Sy; Jinek, Martin; Greene, Eric C.; Doudna, Jennifer A.

    2014-03-01

    The clustered regularly interspaced short palindromic repeats (CRISPR)-associated enzyme Cas9 is an RNA-guided endonuclease that uses RNA-DNA base-pairing to target foreign DNA in bacteria. Cas9-guide RNA complexes are also effective genome engineering agents in animals and plants. Here we use single-molecule and bulk biochemical experiments to determine how Cas9-RNA interrogates DNA to find specific cleavage sites. We show that both binding and cleavage of DNA by Cas9-RNA require recognition of a short trinucleotide protospacer adjacent motif (PAM). Non-target DNA binding affinity scales with PAM density, and sequences fully complementary to the guide RNA but lacking a nearby PAM are ignored by Cas9-RNA. Competition assays provide evidence that DNA strand separation and RNA-DNA heteroduplex formation initiate at the PAM and proceed directionally towards the distal end of the target sequence. Furthermore, PAM interactions trigger Cas9 catalytic activity. These results reveal how Cas9 uses PAM recognition to quickly identify potential target sites while scanning large DNA molecules, and to regulate scission of double-stranded DNA.

  20. Effects of Dimerization of Serratia marcescens Endonuclease on Water Dynamics.

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Chuanying; Beck, Brian W.; Krause, Kurt; Weksberg, Tiffany E.; Pettitt, Bernard M.

    2007-02-15

    The research described in this product was performed in part in the Environmental Molecular Sciences Laboratory, a national scientific user facility sponsored by the Department of Energy's Office of Biological and Environmental Research and located at Pacific Northwest National Laboratory. The dynamics and structure of Serratia marcescens endonuclease and its neighboring solvent are investigated by molecular dynamics (MD). Comparisons are made with structural and biochemical experiments. The dimer form is physiologic and functions more processively than the monomer. We previously found a channel formed by connected clusters of waters from the active site to the dimer interface. Here, we show that dimerization clearly changes correlations in the water structure and dynamics in the active site not seen in the monomer. Our results indicate that water at the active sites of the dimer is less affected compared with bulk solvent than in the monomer where it has much slower characteristic relaxation times. Given that water is a required participant in the reaction, this gives a clear advantage to dimerization in the absence of an apparent ability to use both active sites simultaneously.

  1. Computational study of hydration at the TD damaged site of DNA in complex with repair enzyme T4 endonuclease V

    International Nuclear Information System (INIS)

    Pinak, Miroslav

    2000-02-01

    An analysis of the distribution of water around DNA surface focusing on the role of the distribution of water molecules in the proper recognition of damaged site by repair enzyme T4 Endonuclease V was performed. The native DNA dodecamer, dodecamer with the thymine dimer (TD) and complex of DNA and part of repair enzyme T4 Endonuclease V were examined throughout the 500 ps of molecular dynamics simulation. During simulation the number of water molecules close to the DNA atoms and the residence time were calculated. There is an increase in number of water molecules lying in the close vicinity to TD if compared with those lying close to two native thymines (TT). Densely populated area with water molecules around TD is one of the factors detected by enzyme during scanning process. The residence time was found higher for molecule of the complex and the six water molecules were found occupying the stabile positions between the TD and catalytic center close to atoms P, C3' and N3. These molecules originate water mediated hydrogen bond network that contribute to the stability of complex required for the onset of repair process. (author)

  2. Computational study of hydration at the TD damaged site of DNA in complex with repair enzyme T4 endonuclease V

    Energy Technology Data Exchange (ETDEWEB)

    Pinak, Miroslav [Japan Atomic Energy Research Inst., Tokai, Ibaraki (Japan). Tokai Research Establishment

    2000-02-01

    An analysis of the distribution of water around DNA surface focusing on the role of the distribution of water molecules in the proper recognition of damaged site by repair enzyme T4 Endonuclease V was performed. The native DNA dodecamer, dodecamer with the thymine dimer (TD) and complex of DNA and part of repair enzyme T4 Endonuclease V were examined throughout the 500 ps of molecular dynamics simulation. During simulation the number of water molecules close to the DNA atoms and the residence time were calculated. There is an increase in number of water molecules lying in the close vicinity to TD if compared with those lying close to two native thymines (TT). Densely populated area with water molecules around TD is one of the factors detected by enzyme during scanning process. The residence time was found higher for molecule of the complex and the six water molecules were found occupying the stabile positions between the TD and catalytic center close to atoms P, C3' and N3. These molecules originate water mediated hydrogen bond network that contribute to the stability of complex required for the onset of repair process. (author)

  3. Platinum/yttrium iron garnet inverted structures for spin current transport

    Energy Technology Data Exchange (ETDEWEB)

    Aldosary, Mohammed; Li, Junxue; Tang, Chi; Xu, Yadong; Shi, Jing [Department of Physics and Astronomy and SHINES Energy Frontier Research Center, University of California, Riverside, California 92521 (United States); Zheng, Jian-Guo [Irvine Materials Research Institute, University of California, Irvine, California 92697 (United States); Bozhilov, Krassimir N. [Central Facility for Advanced Microscopy and Microanalysis, University of California, Riverside, California 92521 (United States)

    2016-06-13

    30-80 nm thick yttrium iron garnet (YIG) films are grown by pulsed laser deposition on a 5 nm thick sputtered Pt atop gadolinium gallium garnet substrate (GGG) (110). Upon post-growth rapid thermal annealing, single crystal YIG(110) emerges as if it were epitaxially grown on GGG(110) despite the presence of the intermediate Pt film. The YIG surface shows atomic steps with the root-mean-square roughness of 0.12 nm on flat terraces. Both Pt/YIG and GGG/Pt interfaces are atomically sharp. The resulting YIG(110) films show clear in-plane uniaxial magnetic anisotropy with a well-defined easy axis along 〈001〉 and a peak-to-peak ferromagnetic resonance linewidth of 7.5 Oe at 9.32 GHz, similar to YIG epitaxially grown on GGG. Both spin Hall magnetoresistance and longitudinal spin Seebeck effects in the inverted bilayers indicate excellent Pt/YIG interface quality.

  4. Direct observation of multivalent states and 4 f →3 d charge transfer in Ce-doped yttrium iron garnet thin films

    Science.gov (United States)

    Vasili, H. B.; Casals, B.; Cichelero, R.; Macià, F.; Geshev, J.; Gargiani, P.; Valvidares, M.; Herrero-Martin, J.; Pellegrin, E.; Fontcuberta, J.; Herranz, G.

    2017-07-01

    Due to their large magneto-optic responses, rare-earth-doped yttrium iron garnets, Y3F e5O12 (YIG), are highly regarded for their potential in photonics and magnonics. Here, we consider the case of Ce-doped YIG (Ce-YIG) thin films, in which substitutional C e3 + ions are magnetic because of their 4 f1 ground state. In order to elucidate the impact of Ce substitution on the magnetization of YIG, we have carried out soft x-ray spectroscopy measurements on Ce-YIG films. In particular, we have used the element specificity of x-ray magnetic circular dichroism to extract the individual magnetization curves linked to Ce and Fe ions. Our results show that Ce doping triggers a selective charge transfer from Ce to the Fe tetrahedral sites in the YIG structure. This, in turn, causes a disruption of the electronic and magnetic properties of the parent compound, reducing the exchange coupling between the Ce and Fe magnetic moments and causing atypical magnetic behavior. Our work is relevant for understanding magnetism in rare-earth-doped YIG and, eventually, may enable a quantitative evaluation of the magneto-optical properties of rare-earth incorporation into YIG.

  5. Preparation and characterization of BaFe12O19/Y3Fe5O12 composites

    International Nuclear Information System (INIS)

    Lin, Ying; Kang, Pan; Yang, Haibo; Liu, Miao

    2015-01-01

    Highlights: • BaM/YIG composites were fabricated by microwave sintering method. • A simple sol–gel method was used to synthesize BaM/YIG composite powders. • The BaM phase and YIG phase are well exchange coupled in the composites. • The (BH) max can be much enhanced by the introduction of YIG. - Abstract: BaFe 12 O 19 /Y 3 Fe 5 O 12 (BaM/YIG) composites with giant enhancement of magnetic energy product ((BH) max ) were fabricated by microwave sintering the BaM/YIG composite powders, which were firstly prepared using a simple sol–gel method. The phase composition and surface morphology of the as-synthesized composites were characterized by an X-ray diffractometer and a scanning electron microscopy equipped with energy dispersive X-ray spectroscopy, respectively. The magnetic properties of the composites were investigated by a vibrating sample magnetometer. All the composites show single-phase-like magnetic hysteresis loops. The results reveal the hard phase (BaM) and soft phase (YIG) are well exchange coupled and the introduction of YIG could significantly enhance the remnant magnetization (M r ), coercivity (H c ) and (BH) max of BaM

  6. Small-angle X-ray scattering analysis reveals the ATP-bound monomeric state of the ATPase domain from the homodimeric MutL endonuclease, a GHKL phosphotransferase superfamily protein.

    Science.gov (United States)

    Iino, Hitoshi; Hikima, Takaaki; Nishida, Yuya; Yamamoto, Masaki; Kuramitsu, Seiki; Fukui, Kenji

    2015-05-01

    DNA mismatch repair is an excision system that removes mismatched bases chiefly generated by replication errors. In this system, MutL endonucleases direct the excision reaction to the error-containing strand of the duplex by specifically incising the newly synthesized strand. Both bacterial homodimeric and eukaryotic heterodimeric MutL proteins belong to the GHKL ATPase/kinase superfamily that comprises the N-terminal ATPase and C-terminal dimerization regions. Generally, the GHKL proteins show large ATPase cycle-dependent conformational changes, including dimerization-coupled ATP binding of the N-terminal domain. Interestingly, the ATPase domain of human PMS2, a subunit of the MutL heterodimer, binds ATP without dimerization. The monomeric ATP-bound state of the domain has been thought to be characteristic of heterodimeric GHKL proteins. In this study, we characterized the ATP-bound state of the ATPase domain from the Aquifex aeolicus MutL endonuclease, which is a homodimeric GHKL protein unlike the eukaryotic MutL. Gel filtration, dynamic light scattering, and small-angle X-ray scattering analyses clearly showed that the domain binds ATP in a monomeric form despite its homodimeric nature. This indicates that the uncoupling of dimerization and ATP binding is a common feature among bacterial and eukaryotic MutL endonucleases, which we suggest is closely related to the molecular mechanisms underlying mismatch repair.

  7. Growth and characterization of yttrium iron garnet films on Si substrates by Chemical Solution Deposition (CSD) technique

    Energy Technology Data Exchange (ETDEWEB)

    Guo, Xin; Chen, Ying; Wang, Genshui [Key Laboratory of Inorganic Function Materials and Devices, Shanghai Institute of Ceramics, Chinese Academy of Sciences, University of Chinese Academy of Sciences, 1295 Dingxi Rd., Shanghai 200050 (China); Zhang, Yuanyuan [Key Laboratory of Polar Materials and Devices, Ministry of Education, Department of Electronic Engineering, East China Normal University, 500 Dongchuan Rd., Shanghai 200241 (China); Ge, Jun [Key Laboratory of Inorganic Function Materials and Devices, Shanghai Institute of Ceramics, Chinese Academy of Sciences, University of Chinese Academy of Sciences, 1295 Dingxi Rd., Shanghai 200050 (China); Tang, Xiaodong [Key Laboratory of Polar Materials and Devices, Ministry of Education, Department of Electronic Engineering, East China Normal University, 500 Dongchuan Rd., Shanghai 200241 (China); Ponchel, Freddy; Rémiens, Denis [Institute of Electronics, Microelectronics and Nanotechnology (IEMN)–DOAE, UMR CNRS 8520, Université des Sciences et Technologies de Lille, 59652 Villeneuve d’Ascq Cedex (France); Dong, Xianlin, E-mail: xldong@mail.sic.ac.cn [Key Laboratory of Inorganic Function Materials and Devices, Shanghai Institute of Ceramics, Chinese Academy of Sciences, University of Chinese Academy of Sciences, 1295 Dingxi Rd., Shanghai 200050 (China)

    2016-06-25

    Yttrium Iron Garnet (YIG) films were prepared on Si substrates by Chemical Solution Deposition (CSD) technique using acetic acid and deionized water as solvents. Well-crystallized and crack-free YIG films were obtained when annealed at 750 °C and 850 °C respectively, showing a low surface roughness of several nanometers. When annealed at 750 °C for 30 min, the saturated magnetization (Ms) and coercive field (Hc) of YIG films were 0.121 emu/mm{sup 3} (4πMs = 1.52 kGs) and 7 Oe respectively, which were similar to that prepared by PLD technique. The peak-to-peak linewidth of ferromagnetic resonance (FMR) was 220 Oe at 9.10 GHz. The results demonstrated that CSD was an excellent technique to prepare high quality yttrium iron garnet (YIG) films on silicon, which could provide a lower-cost way for large-scale production on Si-based integrated devices. - Highlights: • The preparation of YIG films by Chemical Solution Deposition are demonstrated. • Well-crystallized and crack-free YIG films can be obtained on Si substrate by CSD. • YIG films can be crystallized in 750 °C with good magnetic performances. • It's beneficial to large-scale production of YIG films on Si integrated devices.

  8. Cloning and analysis of a bifunctional methyltransferase/restriction endonuclease TspGWI, the prototype of a Thermus sp. enzyme family

    Directory of Open Access Journals (Sweden)

    Zylicz-Stachula Agnieszka

    2009-05-01

    Full Text Available Abstract Background Restriction-modification systems are a diverse class of enzymes. They are classified into four major types: I, II, III and IV. We have previously proposed the existence of a Thermus sp. enzyme family, which belongs to type II restriction endonucleases (REases, however, it features also some characteristics of types I and III. Members include related thermophilic endonucleases: TspGWI, TaqII, TspDTI, and Tth111II. Results Here we describe cloning, mutagenesis and analysis of the prototype TspGWI enzyme that recognises the 5'-ACGGA-3' site and cleaves 11/9 nt downstream. We cloned, expressed, and mutagenised the tspgwi gene and investigated the properties of its product, the bifunctional TspGWI restriction/modification enzyme. Since TspGWI does not cleave DNA completely, a cloning method was devised, based on amino acid sequencing of internal proteolytic fragments. The deduced amino acid sequence of the enzyme shares significant sequence similarity with another representative of the Thermus sp. family – TaqII. Interestingly, these enzymes recognise similar, yet different sequences in the DNA. Both enzymes cleave DNA at the same distance, but differ in their ability to cleave single sites and in the requirement of S-adenosylmethionine as an allosteric activator for cleavage. Both the restriction endonuclease (REase and methyltransferase (MTase activities of wild type (wt TspGWI (either recombinant or isolated from Thermus sp. are dependent on the presence of divalent cations. Conclusion TspGWI is a bifunctional protein comprising a tandem arrangement of Type I-like domains; particularly noticeable is the central HsdM-like module comprising a helical domain and a highly conserved S-adenosylmethionine-binding/catalytic MTase domain, containing DPAVGTG and NPPY motifs. TspGWI also possesses an N-terminal PD-(D/EXK nuclease domain related to the corresponding domains in HsdR subunits, but lacks the ATP-dependent translocase module

  9. The Helicobacter pylori HpyAXII restriction–modification system limits exogenous DNA uptake by targeting GTAC sites but shows asymmetric conservation of the DNA methyltransferase and restriction endonuclease components

    Science.gov (United States)

    Humbert, Olivier; Salama, Nina R.

    2008-01-01

    The naturally competent organism Helicobacter pylori encodes a large number of restriction–modification (R–M) systems that consist of a restriction endonuclease and a DNA methyltransferase. R–M systems are not only believed to limit DNA exchange among bacteria but may also have other cellular functions. We report a previously uncharacterized H. pylori type II R–M system, M.HpyAXII/R.HpyAXII. We show that this system targets GTAC sites, which are rare in the H. pylori chromosome but numerous in ribosomal RNA genes. As predicted, this type II R–M system showed attributes of a selfish element. Deletion of the methyltransferase M.HpyAXII is lethal when associated with an active endonuclease R.HpyAXII unless compensated by adaptive mutation or gene amplification. R.HpyAXII effectively restricted both unmethylated plasmid and chromosomal DNA during natural transformation and was predicted to belong to the novel ‘half pipe’ structural family of endonucleases. Analysis of a panel of clinical isolates revealed that R.HpyAXII was functional in a small number of H. pylori strains (18.9%, n = 37), whereas the activity of M.HpyAXII was highly conserved (92%, n = 50), suggesting that GTAC methylation confers a selective advantage to H. pylori. However, M.HpyAXII activity did not enhance H. pylori fitness during stomach colonization of a mouse infection model. PMID:18978016

  10. Comparative studies of the endonucleases from two related Xenopus laevis retrotransposons, Tx1L and Tx2L: target site specificity and evolutionary implications.

    Science.gov (United States)

    Christensen, S; Pont-Kingdon, G; Carroll, D

    2000-01-01

    In the genome of the South African frog, Xenopus laevis, there are two complex families of transposable elements, Tx1 and Tx2, that have identical overall structures, but distinct sequences. In each family there are approximately 1500 copies of an apparent DNA-based element (Tx1D and Tx2D). Roughly 10% of these elements in each family are interrupted by a non-LTR retrotransposon (Tx1L and Tx2L). Each retrotransposon is flanked by a 23-bp target duplication of a specific D element sequence. In earlier work, we showed that the endonuclease domain (Tx1L EN) located in the second open reading frame (ORF2) of Tx1L encodes a protein that makes a single-strand cut precisely at the expected site within its target sequence, supporting the idea that Tx1L is a site-specific retrotransposon. In this study, we express the endonuclease domain of Tx2L (Tx2L EN) and compare the target preferences of the two enzymes. Each endonuclease shows some preference for its cognate target, on the order of 5-fold over the non-cognate target. The observed discrimination is not sufficient, however, to explain the observation that no cross-occupancy is observed - that is, L elements of one family have never been found within D elements of the other family. Possible sources of additional specificity are discussed. We also compare two hypotheses regarding the genome duplication event that led to the contemporary pseudotetraploid character of Xenopus laevis in light of the Tx1L and Tx2L data.

  11. Flap Endonuclease 1 Limits Telomere Fragility on the Leading Strand*

    Science.gov (United States)

    Teasley, Daniel C.; Parajuli, Shankar; Nguyen, Mai; Moore, Hayley R.; Alspach, Elise; Lock, Ying Jie; Honaker, Yuchi; Saharia, Abhishek; Piwnica-Worms, Helen; Stewart, Sheila A.

    2015-01-01

    The existence of redundant replication and repair systems that ensure genome stability underscores the importance of faithful DNA replication. Nowhere is this complexity more evident than in challenging DNA templates, including highly repetitive or transcribed sequences. Here, we demonstrate that flap endonuclease 1 (FEN1), a canonical lagging strand DNA replication protein, is required for normal, complete leading strand replication at telomeres. We find that the loss of FEN1 nuclease activity, but not DNA repair activities, results in leading strand-specific telomere fragility. Furthermore, we show that FEN1 depletion-induced telomere fragility is increased by RNA polymerase II inhibition and is rescued by ectopic RNase H1 expression. These data suggest that FEN1 limits leading strand-specific telomere fragility by processing RNA:DNA hybrid/flap intermediates that arise from co-directional collisions occurring between the replisome and RNA polymerase. Our data reveal the first molecular mechanism for leading strand-specific telomere fragility and the first known role for FEN1 in leading strand DNA replication. Because FEN1 mutations have been identified in human cancers, our findings raise the possibility that unresolved RNA:DNA hybrid structures contribute to the genomic instability associated with cancer. PMID:25922071

  12. Growth of yttrium iron garnet single crystals in Na2O-B2O3 flux system in air

    International Nuclear Information System (INIS)

    Bandyopadhyay, Tarun; Saha, Prasenjit

    1977-01-01

    Detailed studies of growth of yttrium iron garnet (YIG) in the flux system Na 2 O-B 2 O 3 were made to delineate the phase stability regions of YIG and the neighbouring crystalline phases, and a tentative working diagram of Na 2 O-B 2 O 3 -YIG in air was drawn from the results of small batch crystal growth runs and solubility studies. The occurrence of a rather narrow field of YIG crystallization in this system was explained on the basis of relative solubilities of different oxides constituting YIG. Several long-time growth runs with some typical compositions were carried out in this system to evaluate its suitability for bulk growth of YIG crystals. The system, although possessing some inherent advantages, was found to suffer from limitations because the maximum size of the self-nucleated crystals grown hardly exceeded 2.00 mm. (author)

  13. The DNA repair endonuclease XPG interacts directly and functionally with the WRN helicase defective in Werner syndrome

    Energy Technology Data Exchange (ETDEWEB)

    Trego, Kelly S.; Chernikova, Sophia B.; Davalos, Albert R.; Perry, J. Jefferson P.; Finger, L. David; Ng, Cliff; Tsai, Miaw-Sheue; Yannone, Steven M.; Tainer, John A.; Campisi, Judith; Cooper, Priscilla K.

    2011-04-20

    XPG is a structure-specific endonuclease required for nucleotide excision repair (NER). XPG incision defects result in the cancer-prone syndrome xeroderma pigmentosum, whereas truncating mutations of XPG cause the severe postnatal progeroid developmental disorder Cockayne syndrome. We show that XPG interacts directly with WRN protein, which is defective in the premature aging disorder Werner syndrome, and that the two proteins undergo similar sub-nuclear redistribution in S-phase and co-localize in nuclear foci. The co-localization was observed in mid- to late-S-phase, when WRN moves from nucleoli to nuclear foci that have been shown to contain protein markers of both stalled replication forks and telomeric proteins. We mapped the interaction between XPG and WRN to the C-terminal domains of each and show that interaction with the C-terminal domain of XPG strongly stimulates WRN helicase activity. WRN also possesses a competing DNA single-strand annealing activity that, combined with unwinding, has been shown to coordinate regression of model replication forks to form Holliday junction/chicken foot intermediate structures. We tested whether XPG stimulated WRN annealing activity and found that XPG itself has intrinsic strand annealing activity that requires the unstructured R- and C-terminal domains, but not the conserved catalytic core or endonuclease activity. Annealing by XPG is cooperative, rather than additive, with WRN annealing. Taken together, our results suggest a novel function for XPG in S-phase that is at least in part carried out coordinately with WRN, and which may contribute to the severity of the phenotypes that occur upon loss of XPG.

  14. Structural insights of the ssDNA binding site in the multifunctional endonuclease AtBFN2 from Arabidopsis thaliana.

    Directory of Open Access Journals (Sweden)

    Tsung-Fu Yu

    Full Text Available The multi S1/P1 nuclease AtBFN2 (EC 3.1.30.1 encoded by the Arabidopsis thaliana At1g68290 gene is a glycoprotein that digests RNA, ssDNA, and dsDNA. AtBFN2 depends on three zinc ions for cleaving DNA and RNA at 3'-OH to yield 5'-nucleotides. In addition, AtBFN2's enzymatic activity is strongly glycan dependent. Plant Zn(2+-dependent endonucleases present a unique fold, and belong to the Phospholipase C (PLC/P1 nuclease superfamily. In this work, we present the first complete, ligand-free, AtBFN2 crystal structure, along with sulfate, phosphate and ssDNA co-crystal structures. With these, we were able to provide better insight into the glycan structure and possible enzymatic mechanism. In comparison with other nucleases, the AtBFN2/ligand-free and AtBFN2/PO4 models suggest a similar, previously proposed, catalytic mechanism. Our data also confirm that the phosphate and vanadate can inhibit the enzyme activity by occupying the active site. More importantly, the AtBFN2/A5T structure reveals a novel and conserved secondary binding site, which seems to be important for plant Zn(2+-dependent endonucleases. Based on these findings, we propose a rational ssDNA binding model, in which the ssDNA wraps itself around the protein and the attached surface glycan, in turn, reinforces the binding complex.

  15. Next-generation sequencing of multiple individuals per barcoded library by deconvolution of sequenced amplicons using endonuclease fragment analysis

    DEFF Research Database (Denmark)

    Andersen, Jeppe D; Pereira, Vania; Pietroni, Carlotta

    2014-01-01

    The simultaneous sequencing of samples from multiple individuals increases the efficiency of next-generation sequencing (NGS) while also reducing costs. Here we describe a novel and simple approach for sequencing DNA from multiple individuals per barcode. Our strategy relies on the endonuclease...... digestion of PCR amplicons prior to library preparation, creating a specific fragment pattern for each individual that can be resolved after sequencing. By using both barcodes and restriction fragment patterns, we demonstrate the ability to sequence the human melanocortin 1 receptor (MC1R) genes from 72...... individuals using only 24 barcoded libraries....

  16. Differential distribution of a SINE element in the Entamoeba histolytica and Entamoeba dispar genomes: Role of the LINE-encoded endonuclease

    Directory of Open Access Journals (Sweden)

    Gupta Abhishek K

    2011-05-01

    Full Text Available Abstract Background Entamoeba histolytica and Entamoeba dispar are closely related protistan parasites but while E. histolytica can be invasive, E. dispar is completely non pathogenic. Transposable elements constitute a significant portion of the genome in these species; there being three families of LINEs and SINEs. These elements can profoundly influence the expression of neighboring genes. Thus their genomic location can have important phenotypic consequences. A genome-wide comparison of the location of these elements in the E. histolytica and E. dispar genomes has not been carried out. It is also not known whether the retrotransposition machinery works similarly in both species. The present study was undertaken to address these issues. Results Here we extracted all genomic occurrences of full-length copies of EhSINE1 in the E. histolytica genome and matched them with the homologous regions in E. dispar, and vice versa, wherever it was possible to establish synteny. We found that only about 20% of syntenic sites were occupied by SINE1 in both species. We checked whether the different genomic location in the two species was due to differences in the activity of the LINE-encoded endonuclease which is required for nicking the target site. We found that the endonucleases of both species were essentially very similar, both in their kinetic properties and in their substrate sequence specificity. Hence the differential distribution of SINEs in these species is not likely to be influenced by the endonuclease. Further we found that the physical properties of the DNA sequences adjoining the insertion sites were similar in both species. Conclusions Our data shows that the basic retrotransposition machinery is conserved in these sibling species. SINEs may indeed have occupied all of the insertion sites in the genome of the common ancestor of E. histolytica and E. dispar but these may have been subsequently lost from some locations. Alternatively, SINE

  17. Type II restriction endonucleases--a historical perspective and more.

    Science.gov (United States)

    Pingoud, Alfred; Wilson, Geoffrey G; Wende, Wolfgang

    2014-07-01

    This article continues the series of Surveys and Summaries on restriction endonucleases (REases) begun this year in Nucleic Acids Research. Here we discuss 'Type II' REases, the kind used for DNA analysis and cloning. We focus on their biochemistry: what they are, what they do, and how they do it. Type II REases are produced by prokaryotes to combat bacteriophages. With extreme accuracy, each recognizes a particular sequence in double-stranded DNA and cleaves at a fixed position within or nearby. The discoveries of these enzymes in the 1970s, and of the uses to which they could be put, have since impacted every corner of the life sciences. They became the enabling tools of molecular biology, genetics and biotechnology, and made analysis at the most fundamental levels routine. Hundreds of different REases have been discovered and are available commercially. Their genes have been cloned, sequenced and overexpressed. Most have been characterized to some extent, but few have been studied in depth. Here, we describe the original discoveries in this field, and the properties of the first Type II REases investigated. We discuss the mechanisms of sequence recognition and catalysis, and the varied oligomeric modes in which Type II REases act. We describe the surprising heterogeneity revealed by comparisons of their sequences and structures. © The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.

  18. Coupling of the nucleotide incision and 3' {yields} 5' exonuclease activities in Escherichia coli endonuclease IV: Structural and genetic evidences

    Energy Technology Data Exchange (ETDEWEB)

    Golan, Gali [Hebrew University of Jerusalem, Jerusalem 91904 (Israel); Ishchenko, Alexander A. [Groupe Reparation de l' ADN, CNRS UMR 8126, Univ. Paris-Sud, Institut de Cancerologie Gustave Roussy, 39, rue Camille Desmoulins, F-94805 Villejuif Cedex (France); Khassenov, Bekbolat [National Center for Biotechnology, Astana (Kazakhstan); Shoham, Gil, E-mail: gil2@vms.huji.ac.il [Hebrew University of Jerusalem, Jerusalem 91904 (Israel); Saparbaev, Murat K., E-mail: smurat@igr.fr [Groupe Reparation de l' ADN, CNRS UMR 8126, Univ. Paris-Sud, Institut de Cancerologie Gustave Roussy, 39, rue Camille Desmoulins, F-94805 Villejuif Cedex (France)

    2010-03-01

    Aerobic respiration generates reactive oxygen species (ROS) as a by-product of cellular metabolism which can damage DNA. The complex nature of oxidative DNA damage requires actions of several repair pathways. Oxidized DNA bases are substrates for two overlapping pathways: base excision repair (BER) and nucleotide incision repair (NIR). In the BER pathway a DNA glycosylase cleaves the N-glycosylic bond between the abnormal base and deoxyribose, leaving either an abasic site or single-stranded DNA break. Alternatively, in the NIR pathway, an apurinic/apyrimidinic (AP) endonuclease incises duplex DNA 5' next to oxidatively damaged nucleotide. The multifunctional Escherichia coli endonuclease IV (Nfo) is involved in both BER and NIR pathways. Nfo incises duplex DNA 5' of a damaged residue but also possesses an intrinsic 3' {yields} 5' exonuclease activity. Herein, we demonstrate that Nfo-catalyzed NIR and exonuclease activities can generate a single-strand gap at the 5' side of 5,6-dihydrouracil residue. Furthermore, we show that Nfo mutants carrying amino acid substitutions H69A and G149D are deficient in both NIR and exonuclease activities, suggesting that these two functions are genetically linked and governed by the same amino acid residues. The crystal structure of Nfo-H69A mutant reveals the loss of one of the active site zinc atoms (Zn1) and rearrangements of the catalytic site, but no gross changes in the overall enzyme conformation. We hypothesize that these minor changes strongly affect the DNA binding of Nfo. Decreased affinity may lead to a different kinking angle of the DNA helix and this in turn thwart nucleotide incision and exonuclease activities of Nfo mutants but to lesser extent of their AP endonuclease function. Based on the biochemical and genetic data we propose a model where nucleotide incision coupled to 3' {yields} 5' exonuclease activity prevents formation of lethal double-strand breaks when repairing bi

  19. Site- and strand-specific nicking of DNA by fusion proteins derived from MutH and I-SceI or TALE repeats.

    Science.gov (United States)

    Gabsalilow, Lilia; Schierling, Benno; Friedhoff, Peter; Pingoud, Alfred; Wende, Wolfgang

    2013-04-01

    Targeted genome engineering requires nucleases that introduce a highly specific double-strand break in the genome that is either processed by homology-directed repair in the presence of a homologous repair template or by non-homologous end-joining (NHEJ) that usually results in insertions or deletions. The error-prone NHEJ can be efficiently suppressed by 'nickases' that produce a single-strand break rather than a double-strand break. Highly specific nickases have been produced by engineering of homing endonucleases and more recently by modifying zinc finger nucleases (ZFNs) composed of a zinc finger array and the catalytic domain of the restriction endonuclease FokI. These ZF-nickases work as heterodimers in which one subunit has a catalytically inactive FokI domain. We present two different approaches to engineer highly specific nickases; both rely on the sequence-specific nicking activity of the DNA mismatch repair endonuclease MutH which we fused to a DNA-binding module, either a catalytically inactive variant of the homing endonuclease I-SceI or the DNA-binding domain of the TALE protein AvrBs4. The fusion proteins nick strand specifically a bipartite recognition sequence consisting of the MutH and the I-SceI or TALE recognition sequences, respectively, with a more than 1000-fold preference over a stand-alone MutH site. TALE-MutH is a programmable nickase.

  20. Micrococcus luteus correndonucleases. II. Mechanism of action of two endonucleases specific for DNA containing pyrimidine dimers

    International Nuclear Information System (INIS)

    Riazuddin, S.; Grossman, L.

    1977-01-01

    Py--Py correndonucleases I and II from Micrococcus luteus act exclusively on thymine-thymine, cytosine-cytosine, and thymine-cytosine cyclobutyl dimers in DNA, catalyzing incision 5' to the damage and generating 3'-hydroxyl and 5'-phosphoryl termini. Both enzymes initiate excision of pyrimidine dimers in vitro by correxonucleases and DNA polymerase I. The respective incised DNAs, however, differ in their ability to act as substrate for phage T4 polynucleotide ligase or bacterial alkaline phosphatase, suggesting that each endonuclease is specific for a conformationally unique site. The possibility that their respective action generates termini which represent different degrees of single strandedness is suggested by the unequal protection by Escherichia coli binding protein from the hydrolytic action of exonuclease VII

  1. Pre-steady-state fluorescence analysis of damaged DNA transfer from human DNA glycosylases to AP endonuclease APE1.

    Science.gov (United States)

    Kuznetsova, Alexandra A; Kuznetsov, Nikita A; Ishchenko, Alexander A; Saparbaev, Murat K; Fedorova, Olga S

    2014-10-01

    DNA glycosylases remove the modified, damaged or mismatched bases from the DNA by hydrolyzing the N-glycosidic bonds. Some enzymes can further catalyze the incision of a resulting abasic (apurinic/apyrimidinic, AP) site through β- or β,δ-elimination mechanisms. In most cases, the incision reaction of the AP-site is catalyzed by special enzymes called AP-endonucleases. Here, we report the kinetic analysis of the mechanisms of modified DNA transfer from some DNA glycosylases to the AP endonuclease, APE1. The modified DNA contained the tetrahydrofurane residue (F), the analogue of the AP-site. DNA glycosylases AAG, OGG1, NEIL1, MBD4(cat) and UNG from different structural superfamilies were used. We found that all DNA glycosylases may utilise direct protein-protein interactions in the transient ternary complex for the transfer of the AP-containing DNA strand to APE1. We hypothesize a fast "flip-flop" exchange mechanism of damaged and undamaged DNA strands within this complex for monofunctional DNA glycosylases like MBD4(cat), AAG and UNG. Bifunctional DNA glycosylase NEIL1 creates tightly specific complex with DNA containing F-site thereby efficiently competing with APE1. Whereas APE1 fast displaces other bifunctional DNA glycosylase OGG1 on F-site thereby induces its shifts to undamaged DNA regions. Kinetic analysis of the transfer of DNA between human DNA glycosylases and APE1 allows us to elucidate the critical step in the base excision repair pathway. Copyright © 2014 Elsevier B.V. All rights reserved.

  2. Flap Endonuclease 1 Limits Telomere Fragility on the Leading Strand.

    Science.gov (United States)

    Teasley, Daniel C; Parajuli, Shankar; Nguyen, Mai; Moore, Hayley R; Alspach, Elise; Lock, Ying Jie; Honaker, Yuchi; Saharia, Abhishek; Piwnica-Worms, Helen; Stewart, Sheila A

    2015-06-12

    The existence of redundant replication and repair systems that ensure genome stability underscores the importance of faithful DNA replication. Nowhere is this complexity more evident than in challenging DNA templates, including highly repetitive or transcribed sequences. Here, we demonstrate that flap endonuclease 1 (FEN1), a canonical lagging strand DNA replication protein, is required for normal, complete leading strand replication at telomeres. We find that the loss of FEN1 nuclease activity, but not DNA repair activities, results in leading strand-specific telomere fragility. Furthermore, we show that FEN1 depletion-induced telomere fragility is increased by RNA polymerase II inhibition and is rescued by ectopic RNase H1 expression. These data suggest that FEN1 limits leading strand-specific telomere fragility by processing RNA:DNA hybrid/flap intermediates that arise from co-directional collisions occurring between the replisome and RNA polymerase. Our data reveal the first molecular mechanism for leading strand-specific telomere fragility and the first known role for FEN1 in leading strand DNA replication. Because FEN1 mutations have been identified in human cancers, our findings raise the possibility that unresolved RNA:DNA hybrid structures contribute to the genomic instability associated with cancer. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  3. Spin Hall magnetoresistance at high temperatures

    International Nuclear Information System (INIS)

    Uchida, Ken-ichi; Qiu, Zhiyong; Kikkawa, Takashi; Iguchi, Ryo; Saitoh, Eiji

    2015-01-01

    The temperature dependence of spin Hall magnetoresistance (SMR) in Pt/Y 3 Fe 5 O 12 (YIG) bilayer films has been investigated in a high temperature range from room temperature to near the Curie temperature of YIG. The experimental results show that the magnitude of the magnetoresistance ratio induced by the SMR monotonically decreases with increasing the temperature and almost disappears near the Curie temperature. We found that, near the Curie temperature, the temperature dependence of the SMR in the Pt/YIG film is steeper than that of a magnetization curve of the YIG; the critical exponent of the magnetoresistance ratio is estimated to be 0.9. This critical behavior of the SMR is attributed mainly to the temperature dependence of the spin-mixing conductance at the Pt/YIG interface

  4. RECQ5 Helicase Cooperates with MUS81 Endonuclease in Processing Stalled Replication Forks at Common Fragile Sites during Mitosis

    DEFF Research Database (Denmark)

    Di Marco, Stefano; Hasanova, Zdenka; Kanagaraj, Radhakrishnan

    2017-01-01

    The MUS81-EME1 endonuclease cleaves late replication intermediates at common fragile sites (CFSs) during early mitosis to trigger DNA-repair synthesis that ensures faithful chromosome segregation. Here, we show that these DNA transactions are promoted by RECQ5 DNA helicase in a manner dependent...... on its Ser727 phosphorylation by CDK1. Upon replication stress, RECQ5 associates with CFSs in early mitosis through its physical interaction with MUS81 and promotes MUS81-dependent mitotic DNA synthesis. RECQ5 depletion or mutational inactivation of its ATP-binding site, RAD51-interacting domain...

  5. Primary processing of CRISPR RNA by the endonuclease Cas6 in Staphylococcus epidermidis.

    Science.gov (United States)

    Wakefield, Noelle; Rajan, Rakhi; Sontheimer, Erik J

    2015-10-07

    In many bacteria and archaea, an adaptive immune system (CRISPR-Cas) provides immunity against foreign genetic elements. This system uses CRISPR RNAs (crRNAs) derived from the CRISPR array, along with CRISPR-associated (Cas) proteins, to target foreign nucleic acids. In most CRISPR systems, endonucleolytic processing of crRNA precursors (pre-crRNAs) is essential for the pathway. Here we study the Cas6 endonuclease responsible for crRNA processing in the Type III-A CRISPR-Cas system from Staphylococcus epidermidis RP62a, a model for Type III-A CRISPR-Cas systems, and define substrate requirements for SeCas6 activity. We find that SeCas6 is necessary and sufficient for full-length crRNA biogenesis in vitro, and that it relies on both sequence and stem-loop structure in the 3' half of the CRISPR repeat for recognition and processing. Copyright © 2015 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

  6. Computational Characterization of Small Molecules Binding to the Human XPF Active Site and Virtual Screening to Identify Potential New DNA Repair Inhibitors Targeting the ERCC1-XPF Endonuclease

    Directory of Open Access Journals (Sweden)

    Francesco Gentile

    2018-04-01

    Full Text Available The DNA excision repair protein ERCC-1-DNA repair endonuclease XPF (ERCC1-XPF is a heterodimeric endonuclease essential for the nucleotide excision repair (NER DNA repair pathway. Although its activity is required to maintain genome integrity in healthy cells, ERCC1-XPF can counteract the effect of DNA-damaging therapies such as platinum-based chemotherapy in cancer cells. Therefore, a promising approach to enhance the effect of these therapies is to combine their use with small molecules, which can inhibit the repair mechanisms in cancer cells. Currently, there are no structures available for the catalytic site of the human ERCC1-XPF, which performs the metal-mediated cleavage of a DNA damaged strand at 5′. We adopted a homology modeling strategy to build a structural model of the human XPF nuclease domain which contained the active site and to extract dominant conformations of the domain using molecular dynamics simulations followed by clustering of the trajectory. We investigated the binding modes of known small molecule inhibitors targeting the active site to build a pharmacophore model. We then performed a virtual screening of the ZINC Is Not Commercial 15 (ZINC15 database to identify new ERCC1-XPF endonuclease inhibitors. Our work provides structural insights regarding the binding mode of small molecules targeting the ERCC1-XPF active site that can be used to rationally optimize such compounds. We also propose a set of new potential DNA repair inhibitors to be considered for combination cancer therapy strategies.

  7. Reflection and Transmission Coefficient of Yttrium Iron Garnet Filled Polyvinylidene Fluoride Composite Using Rectangular Waveguide at Microwave Frequencies

    Science.gov (United States)

    Soleimani, Hassan; Abbas, Zulkifly; Yahya, Noorhana; Shameli, Kamyar; Soleimani, Hojjatollah; Shabanzadeh, Parvaneh

    2012-01-01

    The sol-gel method was carried out to synthesize nanosized Yttrium Iron Garnet (YIG). The nanomaterials with ferrite structure were heat-treated at different temperatures from 500 to 1000 °C. The phase identification, morphology and functional groups of the prepared samples were characterized by powder X-ray diffraction (PXRD), scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FT-IR), respectively. The YIG ferrite nanopowder was composited with polyvinylidene fluoride (PVDF) by a solution casting method. The magnitudes of reflection and transmission coefficients of PVDF/YIG containing 6, 10 and 13% YIG, respectively, were measured using rectangular waveguide in conjunction with a microwave vector network analyzer (VNA) in X-band frequencies. The results indicate that the presence of YIG in polymer composites causes an increase in reflection coefficient and decrease in transmission coefficient of the polymer. PMID:22942718

  8. High-resolution structural characterization and magnetic properties of epitaxial Ce-doped yttrium iron garnet thin films

    Science.gov (United States)

    Li, Zhong; Vikram Singh, Amit; Rastogi, Ankur; Gazquez, Jaume; Borisevich, Albina Y.; Mishra, Rohan; Gupta, Arunava

    2017-07-01

    Thin films of magnetic garnet materials, e.g. yttrium iron garnet (Y3Fe5O12, YIG), are useful for a variety of applications including microwave integrated circuits and spintronics. Substitution of rare earth ions, such as cerium, is known to enhance the magneto-optic Kerr effect (MOKE) as compared to pure YIG. Thin films of Ce0.75Y2.25Fe5O12 (Ce:YIG) have been grown using the pulsed laser deposition (PLD) technique and their crystal structure examined using high resolution scanning transmission electron microscopy. Homogeneous substitution of Ce in YIG, without oxidation to form a separate CeO2 phase, can be realized in a narrow process window with resulting enhancement of the MOKE signal. The thermally generated signal due to spin Seebeck effect for the optimally doped Ce:YIG films has also been investigated.

  9. Activation of apurinic/apyrimidinic endonuclease in human cells by reactive oxygen species and its correlation with their adaptive response to genotoxicity of free radicals

    Science.gov (United States)

    Ramana, Chilakamarti V.; Boldogh, Istvan; Izumi, Tadahide; Mitra, Sankar

    1998-01-01

    Apurinic/apyrimidinic (AP) endonuclease (APE; EC 4.2.99.18) plays a central role in repair of DNA damage due to reactive oxygen species (ROS) because its DNA 3′-phosphoesterase activity removes 3′ blocking groups in DNA that are generated by DNA glycosylase/AP-lyases during removal of oxidized bases and by direct ROS reaction with DNA. The major human APE (APE-1) gene is activated selectively by sublethal levels of a variety of ROS and ROS generators, including ionizing radiation, but not by other genotoxicants—e.g., UV light and alkylating agents. Increased expression of APE mRNA and protein was observed both in the HeLa S3 tumor line and in WI 38 primary fibroblasts, and it was accompanied by translocation of the endonuclease to the nucleus. ROS-treated cells showed a significant increase in resistance to the cytotoxicity of such ROS generators as H2O2 and bleomycin, but not to UV light. This “adaptive response” appears to result from enhanced repair of cytotoxic DNA lesions due to an increased activity of APE-1, which may be limiting in the base excision repair process for ROS-induced toxic lesions. PMID:9560228

  10. [Endonuclease modified comet assay for oxidative DNA damage induced by detection of genetic toxicants].

    Science.gov (United States)

    Zhao, Jian; Li, Hongli; Zhai, Qingfeng; Qiu, Yugang; Niu, Yong; Dai, Yufei; Zheng, Yuxin; Duan, Huawei

    2014-03-01

    The aim of this study was to investigate the use of the lesion-specific endonucleases-modified comet assay for analysis of DNA oxidation in cell lines. DNA breaks and oxidative damage were evaluated by normal alkaline and formamidopyrimidine-DNA-glycosylase (FPG) modified comet assays. Cytotoxicity were assessed by MTT method. The human bronchial epithelial cell (16HBE) were treated with benzo (a) pyrene (B(a)P), methyl methanesulfonate (MMS), colchicine (COL) and vincristine (VCR) respectively, and the dose is 20 µmol/L, 25 mg/ml, 5 mg/L and 0.5 mg/L for 24 h, respectively. Oxidative damage was also detected by levels of reactive oxygen species in treated cells. Four genotoxicants give higher cytotoxicity and no significant changes on parameters of comet assay treated by enzyme buffer. Cell survival rate were (59.69 ± 2.60) %, (54.33 ± 2.81) %, (53.11 ± 4.00) %, (51.43 ± 3.92) % in four groups, respectively. There was the direct DNA damage induced by test genotoxicants presented by tail length, Olive tail moment (TM) and tail DNA (%) in the comet assay. The presence of FPG in the assays increased DNA migration in treated groups when compared to those without it, and the difference was statistically significant which indicated that the clastogen and aneugen could induce oxidative damage in DNA strand. In the three parameters, the Olive TM was changed most obviously after genotoxicants treatment. In the contrast group, the Olive TM of B(a) P,MMS, COL,VCR in the contrast groups were 22.99 ± 17.33, 31.65 ± 18.86, 19.86 ± 9.56 and 17.02 ± 9.39, respectively, after dealing with the FPG, the Olive TM were 34.50 ± 17.29, 43.80 ± 10.06, 33.10 ± 12.38, 28.60 ± 10.53, increased by 58.94%, 38.48%, 66.86% and 68.21%, respectively (t value was 3.91, 3.89, 6.66 and 3.87, respectively, and all P comet assay appears more specific for detecting oxidative DNA damage induced by genotoxicants exposure, and the application of comet assay will be expanded. The endonuclease

  11. Hold your horSSEs: controlling structure-selective endonucleases MUS81 and Yen1/GEN1.

    Science.gov (United States)

    Blanco, Miguel G; Matos, Joao

    2015-01-01

    Repair of DNA lesions through homologous recombination promotes the establishment of stable chromosomal interactions. Multiple helicases, topoisomerases and structure-selective endonucleases (SSEs) act upon recombining joint molecules (JMs) to disengage chromosomal connections and safeguard chromosome segregation. Recent studies on two conserved SSEs - MUS81 and Yen1/GEN1- uncovered multiple layers of regulation that operate to carefully tailor JM-processing according to specific cellular needs. Temporal restriction of SSE function imposes a hierarchy in pathway usage that ensures efficient JM-processing while minimizing reciprocal exchanges between the recombining DNAs. Whereas a conserved strategy of fine-tuning SSE functions exists in different model systems, the precise molecular mechanisms to implement it appear to be significantly different. Here, we summarize the current knowledge on the cellular switches that are in place to control MUS81 and Yen1/GEN1 functions.

  12. Single-molecule FRET unveils induced-fit mechanism for substrate selectivity in flap endonuclease 1

    KAUST Repository

    Rashid, Fahad

    2017-02-23

    Human flap endonuclease 1 (FEN1) and related structure-specific 5\\'nucleases precisely identify and incise aberrant DNA structures during replication, repair and recombination to avoid genomic instability. Yet, it is unclear how the 5\\'nuclease mechanisms of DNA distortion and protein ordering robustly mediate efficient and accurate substrate recognition and catalytic selectivity. Here, single-molecule sub-millisecond and millisecond analyses of FEN1 reveal a protein-DNA induced-fit mechanism that efficiently verifies substrate and suppresses off-target cleavage. FEN1 sculpts DNA with diffusion-limited kinetics to test DNA substrate. This DNA distortion mutually \\'locks\\' protein and DNA conformation and enables substrate verification with extreme precision. Strikingly, FEN1 never misses cleavage of its cognate substrate while blocking probable formation of catalytically competent interactions with noncognate substrates and fostering their pre-incision dissociation. These findings establish FEN1 has practically perfect precision and that separate control of induced-fit substrate recognition sets up the catalytic selectivity of the nuclease active site for genome stability.

  13. Small molecule inhibitors uncover synthetic genetic interactions of human flap endonuclease 1 (FEN1 with DNA damage response genes.

    Directory of Open Access Journals (Sweden)

    Thomas A Ward

    Full Text Available Flap endonuclease 1 (FEN1 is a structure selective endonuclease required for proficient DNA replication and the repair of DNA damage. Cellularly active inhibitors of this enzyme have previously been shown to induce a DNA damage response and, ultimately, cell death. High-throughput screens of human cancer cell-lines identify colorectal and gastric cell-lines with microsatellite instability (MSI as enriched for cellular sensitivity to N-hydroxyurea series inhibitors of FEN1, but not the PARP inhibitor olaparib or other inhibitors of the DNA damage response. This sensitivity is due to a synthetic lethal interaction between FEN1 and MRE11A, which is often mutated in MSI cancers through instabilities at a poly(T microsatellite repeat. Disruption of ATM is similarly synthetic lethal with FEN1 inhibition, suggesting that disruption of FEN1 function leads to the accumulation of DNA double-strand breaks. These are likely a result of the accumulation of aberrant replication forks, that accumulate as a consequence of a failure in Okazaki fragment maturation, as inhibition of FEN1 is toxic in cells disrupted for the Fanconi anemia pathway and post-replication repair. Furthermore, RAD51 foci accumulate as a consequence of FEN1 inhibition and the toxicity of FEN1 inhibitors increases in cells disrupted for the homologous recombination pathway, suggesting a role for homologous recombination in the resolution of damage induced by FEN1 inhibition. Finally, FEN1 appears to be required for the repair of damage induced by olaparib and cisplatin within the Fanconi anemia pathway, and may play a role in the repair of damage associated with its own disruption.

  14. Observation of the spin Peltier effect for magnetic insulators.

    Science.gov (United States)

    Flipse, J; Dejene, F K; Wagenaar, D; Bauer, G E W; Ben Youssef, J; van Wees, B J

    2014-07-11

    We report the observation of the spin Peltier effect (SPE) in the ferrimagnetic insulator yttrium iron garnet (YIG), i.e., a heat current generated by a spin current flowing through a platinum (Pt)|YIG interface. The effect can be explained by the spin transfer torque that transforms the spin current in the Pt into a magnon current in the YIG. Via magnon-phonon interactions the magnetic fluctuations modulate the phonon temperature that is detected by a thermopile close to the interface. By finite-element modeling we verify the reciprocity between the spin Peltier and spin Seebeck effect. The observed strong coupling between thermal magnons and phonons in YIG is attractive for nanoscale cooling techniques.

  15. Modulation of pure spin currents with a ferromagnetic insulator

    Science.gov (United States)

    Villamor, Estitxu; Isasa, Miren; Vélez, Saül; Bedoya-Pinto, Amilcar; Vavassori, Paolo; Hueso, Luis E.; Bergeret, F. Sebastián; Casanova, Fèlix

    2015-01-01

    We propose and demonstrate spin manipulation by magnetically controlled modulation of pure spin currents in cobalt/copper lateral spin valves, fabricated on top of the magnetic insulator Y3F e5O12 (YIG). The direction of the YIG magnetization can be controlled by a small magnetic field. We observe a clear modulation of the nonlocal resistance as a function of the orientation of the YIG magnetization with respect to the polarization of the spin current. Such a modulation can only be explained by assuming a finite spin-mixing conductance at the Cu/YIG interface, as it follows from the solution of the spin-diffusion equation. These results open a path towards the development of spin logics.

  16. Nonlocal magnon spin transport in yttrium iron garnet with tantalum and platinum spin injection/detection electrodes

    Science.gov (United States)

    Liu, J.; Cornelissen, L. J.; Shan, J.; van Wees, B. J.; Kuschel, T.

    2018-06-01

    We study the magnon spin transport in the magnetic insulator yttrium iron garnet (YIG) in a nonlocal experiment and compare the magnon spin excitation and detection for the heavy metal paramagnetic electrodes platinum (Pt|YIG|Pt) and tantalum (Ta|YIG|Ta). The electrical injection and detection processes rely on the (inverse) spin Hall effect in the heavy metals and the conversion between the electron spin and magnon spin at the heavy metal|YIG interface. Pt and Ta possess opposite signs of the spin Hall angle. Furthermore, their heterostructures with YIG have different interface properties, i.e. spin mixing conductances. By varying the distance between injector and detector, the magnon spin transport is studied. Using a circuit model based on the diffusion-relaxation transport theory, a similar magnon relaxation length of  ∼10 μm was extracted from both Pt and Ta devices. By changing the injector and detector material from Pt to Ta, the influence of interface properties on the magnon spin transport has been observed. For Ta devices on YIG the spin mixing conductance is reduced compared with Pt devices, which is quantitatively consistent when comparing the dependence of the nonlocal signal on the injector-detector distance with the prediction from the circuit model.

  17. Evaluation of an Internally Controlled Multiplex Tth Endonuclease Cleavage Loop-Mediated Isothermal Amplification (TEC-LAMP Assay for the Detection of Bacterial Meningitis Pathogens

    Directory of Open Access Journals (Sweden)

    Owen Higgins

    2018-02-01

    Full Text Available Bacterial meningitis infection is a leading global health concern for which rapid and accurate diagnosis is essential to reduce associated morbidity and mortality. Loop-mediated isothermal amplification (LAMP offers an effective low-cost diagnostic approach; however, multiplex LAMP is difficult to achieve, limiting its application. We have developed novel real-time multiplex LAMP technology, TEC-LAMP, using Tth endonuclease IV and a unique LAMP primer/probe. This study evaluates the analytical specificity, limit of detection (LOD and clinical application of an internally controlled multiplex TEC-LAMP assay for detection of leading bacterial meningitis pathogens: Streptococcus pneumoniae, Neisseria meningitidis and Haemophilus influenzae. Analytical specificities were established by testing 168 bacterial strains, and LODs were determined using Probit analysis. The TEC-LAMP assay was 100% specific, with LODs for S. pneumoniae, N. meningitidis and H. influenzae of 39.5, 17.3 and 25.9 genome copies per reaction, respectively. Clinical performance was evaluated by testing 65 archived PCR-positive samples. Compared to singleplex real-time PCR, the multiplex TEC-LAMP assay demonstrated diagnostic sensitivity and specificity of 92.3% and 100%, respectively. This is the first report of a single-tube internally controlled multiplex LAMP assay for bacterial meningitis pathogen detection, and the first report of Tth endonuclease IV incorporation into nucleic acid amplification diagnostic technology.

  18. Evaluation of an Internally Controlled Multiplex Tth Endonuclease Cleavage Loop-Mediated Isothermal Amplification (TEC-LAMP) Assay for the Detection of Bacterial Meningitis Pathogens

    Science.gov (United States)

    Clancy, Eoin; Cormican, Martin; Boo, Teck Wee; Cunney, Robert

    2018-01-01

    Bacterial meningitis infection is a leading global health concern for which rapid and accurate diagnosis is essential to reduce associated morbidity and mortality. Loop-mediated isothermal amplification (LAMP) offers an effective low-cost diagnostic approach; however, multiplex LAMP is difficult to achieve, limiting its application. We have developed novel real-time multiplex LAMP technology, TEC-LAMP, using Tth endonuclease IV and a unique LAMP primer/probe. This study evaluates the analytical specificity, limit of detection (LOD) and clinical application of an internally controlled multiplex TEC-LAMP assay for detection of leading bacterial meningitis pathogens: Streptococcus pneumoniae, Neisseria meningitidis and Haemophilus influenzae. Analytical specificities were established by testing 168 bacterial strains, and LODs were determined using Probit analysis. The TEC-LAMP assay was 100% specific, with LODs for S. pneumoniae, N. meningitidis and H. influenzae of 39.5, 17.3 and 25.9 genome copies per reaction, respectively. Clinical performance was evaluated by testing 65 archived PCR-positive samples. Compared to singleplex real-time PCR, the multiplex TEC-LAMP assay demonstrated diagnostic sensitivity and specificity of 92.3% and 100%, respectively. This is the first report of a single-tube internally controlled multiplex LAMP assay for bacterial meningitis pathogen detection, and the first report of Tth endonuclease IV incorporation into nucleic acid amplification diagnostic technology. PMID:29425124

  19. Low-relaxation spin waves in laser-molecular-beam epitaxy grown nanosized yttrium iron garnet films

    Energy Technology Data Exchange (ETDEWEB)

    Lutsev, L. V., E-mail: l-lutsev@mail.ru; Korovin, A. M.; Bursian, V. E.; Gastev, S. V.; Fedorov, V. V.; Suturin, S. M.; Sokolov, N. S. [Ioffe Physical-Technical Institute, Russian Academy of Sciences, 194021 St. Petersburg (Russian Federation)

    2016-05-02

    Synthesis of nanosized yttrium iron garnet (Y{sub 3}Fe{sub 5}O{sub 12}, YIG) films followed by the study of ferromagnetic resonance (FMR) and spin wave propagation in these films is reported. The YIG films were grown on gadolinium gallium garnet substrates by laser molecular beam epitaxy. It has been shown that spin waves propagating in YIG deposited at 700 °C have low damping. At the frequency of 3.29 GHz, the spin-wave damping parameter is less than 3.6 × 10{sup −5}. Magnetic inhomogeneities of the YIG films give the main contribution to the FMR linewidth. The contribution of the relaxation processes to the FMR linewidth is as low as 1.2%.

  20. Early endonuclease-mediated evasion of RNA sensing ensures efficient coronavirus replication.

    Directory of Open Access Journals (Sweden)

    Eveline Kindler

    2017-02-01

    Full Text Available Coronaviruses are of veterinary and medical importance and include highly pathogenic zoonotic viruses, such as SARS-CoV and MERS-CoV. They are known to efficiently evade early innate immune responses, manifesting in almost negligible expression of type-I interferons (IFN-I. This evasion strategy suggests an evolutionary conserved viral function that has evolved to prevent RNA-based sensing of infection in vertebrate hosts. Here we show that the coronavirus endonuclease (EndoU activity is key to prevent early induction of double-stranded RNA (dsRNA host cell responses. Replication of EndoU-deficient coronaviruses is greatly attenuated in vivo and severely restricted in primary cells even during the early phase of the infection. In macrophages we found immediate induction of IFN-I expression and RNase L-mediated breakdown of ribosomal RNA. Accordingly, EndoU-deficient viruses can retain replication only in cells that are deficient in IFN-I expression or sensing, and in cells lacking both RNase L and PKR. Collectively our results demonstrate that the coronavirus EndoU efficiently prevents simultaneous activation of host cell dsRNA sensors, such as Mda5, OAS and PKR. The localization of the EndoU activity at the site of viral RNA synthesis-within the replicase complex-suggests that coronaviruses have evolved a viral RNA decay pathway to evade early innate and intrinsic antiviral host cell responses.

  1. Chemical solution synthesis and ferromagnetic resonance of epitaxial thin films of yttrium iron garnet

    Science.gov (United States)

    Lucas, Irene; Jiménez-Cavero, Pilar; Vila-Fungueiriño, J. M.; Magén, Cesar; Sangiao, Soraya; de Teresa, José Maria; Morellón, Luis; Rivadulla, Francisco

    2017-12-01

    We report the fabrication of epitaxial Y3F e5O12 (YIG) thin films on G d3G a5O12 (111) using a chemical solution method. Cubic YIG is a ferrimagnetic material at room temperature, with excellent magneto-optical properties, high electrical resistivity, and a very narrow ferromagnetic resonance, which makes it particularly suitable for applications in filters and resonators at microwave frequencies. But these properties depend on the precise stoichiometry and distribution of F e3 + ions among the octahedral/tetrahedral sites of a complex structure, which hampered the production of high-quality YIG thin films by affordable chemical methods. Here we report the chemical solution synthesis of YIG thin films, with excellent chemical, crystalline, and magnetic homogeneity. The films show a very narrow ferromagnetic resonance (long spin relaxation time), comparable to that obtained from high-vacuum physical deposition methods. These results demonstrate that chemical methods can compete to develop nanometer-thick YIG films with the quality required for spintronic devices and other high-frequency applications.

  2. Positioning the 5'-flap junction in the active site controls the rate of flap endonuclease-1-catalyzed DNA cleavage

    KAUST Repository

    Song, Bo

    2018-02-09

    Flap endonucleases catalyze cleavage of single-stranded DNA flaps formed during replication, repair and recombination, and are therefore essential for genome processing and stability. Recent crystal structures of DNA-bound human flap endonuclease (hFEN1) offer new insights into how conformational changes in the DNA and hFEN1 may facilitate the reaction mechanism. For example, previous biochemical studies of DNA conformation performed under non-catalytic conditions with Ca2+ have suggested that base unpairing at the 5\\'-flap:template junction is an important step in the reaction, but the new structural data suggest otherwise. To clarify the role of DNA changes in the kinetic mechanism, we measured a series of transient steps - from substrate binding to product release - during the hFEN1-catalyzed reaction in the presence of Mg2+. We found that while hFEN1 binds and bends DNA at a fast, diffusion-limited rate, much slower Mg2+-dependent conformational changes in DNA around the active site are subsequently necessary and rate-limiting for 5\\'-flap cleavage. These changes are reported overall by fluorescence of 2-aminopurine at the 5\\'-flap:template junction, indicating that local DNA distortion (e.g., disruption of base stacking observed in structures), associated with positioning the 5\\'-flap scissile phosphodiester bond in the hFEN1 active site, controls catalysis. hFEN1 residues with distinct roles in the catalytic mechanism, including those binding metal ions (Asp-34, Asp-181), steering the 5\\'-flap through the active site and binding the scissile phosphate (Lys-93, Arg-100), and stacking against the base 5\\' to the scissile phosphate (Tyr-40), all contribute to these rate-limiting conformational changes, ensuring efficient and specific cleavage of 5\\'-flaps.

  3. Haplotype-based case-control study on human apurinic/apyrimidinic endonuclease 1/redox effector factor-1 gene and essential hypertension.

    Science.gov (United States)

    Naganuma, Takahiro; Nakayama, Tomohiro; Sato, Naoyuki; Fu, Zhenyan; Soma, Masayoshi; Yamaguchi, Mai; Shimodaira, Masanori; Aoi, Noriko; Usami, Ron

    2010-02-01

    Oxidative DNA damage is involved in the pathophysiology of essential hypertension (EH), which is a multifactorial disorder. Apurinic/apyrimidinic endonuclease 1/redox effector factor-1 (APE1/REF-1) is an essential endonuclease in the base excision repair pathway of oxidatively damaged DNA, in addition to having reducing properties that promote the binding of redox-sensitive transcription factors. Blood pressure in APE1/REF-1-knockout mice is reported to be significantly higher than in wild-type mice. The aim of this study was to investigate the relationship between EH and the human APE1/REF-1 gene through a haplotype-based case-control study using single-nucleotide polymorphisms (SNPs). We selected five SNPs in the human APE1/REF-1 gene (rs1760944, rs3136814, rs17111967, rs3136817, and rs1130409), and performed case-control studies in 265 EH patients and 266 age-matched normotensive (NT) subjects. rs17111967 was found to show nonheterogeneity among Japanese subjects. There were no significant differences in the overall distribution of genotypes or alleles for each SNP between EH and NT groups. In the overall distribution of the haplotype-based case-control study constructed based on rs1760944, rs3136817, and rs1130409, the frequency of the G-T-T haplotype was significantly higher in the EH group than in the NT group (2.1% vs. 0.0%, P = 0.001). Multiple logistic regression analysis also revealed significant differences for the G-T-T haplotype, even after adjustment for confounding factors (OR = 8.600, 95% CI: 1.073-68.951, P = 0.043). Based on the present results, the G-T-T haplotype appears to be a genetic marker of EH, and the APE1/REF-1 gene appears to be a susceptibility gene for EH.

  4. Positioning the 5'-flap junction in the active site controls the rate of flap endonuclease-1-catalyzed DNA cleavage

    KAUST Repository

    Song, Bo; Hamdan, Samir; Hingorani, Manju M

    2018-01-01

    Flap endonucleases catalyze cleavage of single-stranded DNA flaps formed during replication, repair and recombination, and are therefore essential for genome processing and stability. Recent crystal structures of DNA-bound human flap endonuclease (hFEN1) offer new insights into how conformational changes in the DNA and hFEN1 may facilitate the reaction mechanism. For example, previous biochemical studies of DNA conformation performed under non-catalytic conditions with Ca2+ have suggested that base unpairing at the 5'-flap:template junction is an important step in the reaction, but the new structural data suggest otherwise. To clarify the role of DNA changes in the kinetic mechanism, we measured a series of transient steps - from substrate binding to product release - during the hFEN1-catalyzed reaction in the presence of Mg2+. We found that while hFEN1 binds and bends DNA at a fast, diffusion-limited rate, much slower Mg2+-dependent conformational changes in DNA around the active site are subsequently necessary and rate-limiting for 5'-flap cleavage. These changes are reported overall by fluorescence of 2-aminopurine at the 5'-flap:template junction, indicating that local DNA distortion (e.g., disruption of base stacking observed in structures), associated with positioning the 5'-flap scissile phosphodiester bond in the hFEN1 active site, controls catalysis. hFEN1 residues with distinct roles in the catalytic mechanism, including those binding metal ions (Asp-34, Asp-181), steering the 5'-flap through the active site and binding the scissile phosphate (Lys-93, Arg-100), and stacking against the base 5' to the scissile phosphate (Tyr-40), all contribute to these rate-limiting conformational changes, ensuring efficient and specific cleavage of 5'-flaps.

  5. Sequential and Multistep Substrate Interrogation Provides the Scaffold for Specificity in Human Flap Endonuclease 1

    KAUST Repository

    Sobhy, M.; Joudeh, L.; Huang, X.; Takahashi, Masateru; Hamdan, S.

    2013-01-01

    Human flap endonuclease 1 (FEN1), one of the structure-specific 5' nucleases, is integral in replication, repair, and recombination of cellular DNA. The 5' nucleases share significant unifying features yet cleave diverse substrates at similar positions relative to 5' end junctions. Using single-molecule Förster resonance energy transfer, we find a multistep mechanism that verifies all substrate features before inducing the intermediary-DNA bending step that is believed to unify 5' nuclease mechanisms. This is achieved by coordinating threading of the 5' flap of a nick junction into the conserved capped-helical gateway, overseeing the active site, and bending by binding at the base of the junction. We propose that this sequential and multistep substrate recognition process allows different 5' nucleases to recognize different substrates and restrict the induction of DNA bending to the last common step. Such mechanisms would also ensure the protection ofDNA junctions from nonspecific bending and cleavage. 2013 The Authors.

  6. Sequential and Multistep Substrate Interrogation Provides the Scaffold for Specificity in Human Flap Endonuclease 1

    KAUST Repository

    Sobhy, M.

    2013-06-06

    Human flap endonuclease 1 (FEN1), one of the structure-specific 5\\' nucleases, is integral in replication, repair, and recombination of cellular DNA. The 5\\' nucleases share significant unifying features yet cleave diverse substrates at similar positions relative to 5\\' end junctions. Using single-molecule Förster resonance energy transfer, we find a multistep mechanism that verifies all substrate features before inducing the intermediary-DNA bending step that is believed to unify 5\\' nuclease mechanisms. This is achieved by coordinating threading of the 5\\' flap of a nick junction into the conserved capped-helical gateway, overseeing the active site, and bending by binding at the base of the junction. We propose that this sequential and multistep substrate recognition process allows different 5\\' nucleases to recognize different substrates and restrict the induction of DNA bending to the last common step. Such mechanisms would also ensure the protection ofDNA junctions from nonspecific bending and cleavage. 2013 The Authors.

  7. Home safe home: Evaluation of a childhood home safety program.

    Science.gov (United States)

    Stewart, Tanya Charyk; Clark, Andrew; Gilliland, Jason; Miller, Michael R; Edwards, Jane; Haidar, Tania; Batey, Brandon; Vogt, Kelly N; Parry, Neil G; Fraser, Douglas D; Merritt, Neil

    2016-09-01

    The London Health Sciences Centre Home Safety Program (HSP) provides safety devices, education, a safety video, and home safety checklist to all first-time parents for the reduction of childhood home injuries. The objective of this study was to evaluate the HSP for the prevention of home injuries in children up to 2 years of age. A program evaluation was performed with follow-up survey, along with an interrupted time series analysis of emergency department (ED) visits for home injuries 5 years before (2007-2013) and 2 years after (2013-2015) implementation. Spatial analysis of ED visits was undertaken to assess differences in home injury rates by dissemination areas controlling differences in socioeconomic status (i.e., income, education, and lone-parent status) at the neighborhood level. A total of 3,458 first-time parents participated in the HSP (a 74% compliance rate). Of these, 20% (n = 696) of parents responded to our questionnaire, with 94% reporting the program to be useful (median, 6; interquartile range, 2 on a 7-point Likert scale) and 81% learning new strategies for preventing home injuries. The median age of the respondent's babies were 12 months (interquartile range, 1). The home safety check list was used by 87% of respondents to identify hazards in their home, with 95% taking action to minimize the risk. The time series analysis demonstrated a significant decline in ED visits for home injuries in toddlers younger than2 years of age after HSP implementation. The declines in ED visits for home injuries remained significant over and above each socioeconomic status covariate. Removing hazards, supervision, and installing safety devices are key facilitators in the reduction of home injuries. Parents found the HSP useful to identify hazards, learn new strategies, build confidence, and provide safety products. Initial finding suggests that the program is effective in reducing home injuries in children up to 2 years of age. Therapeutic/care management study

  8. Host Factors Influencing the Retrohoming Pathway of Group II Intron RmInt1, Which Has an Intron-Encoded Protein Naturally Devoid of Endonuclease Activity.

    Directory of Open Access Journals (Sweden)

    Rafael Nisa-Martínez

    Full Text Available Bacterial group II introns are self-splicing catalytic RNAs and mobile retroelements that have an open reading frame encoding an intron-encoded protein (IEP with reverse transcriptase (RT and RNA splicing or maturase activity. Some IEPs carry a DNA endonuclease (En domain, which is required to cleave the bottom strand downstream from the intron-insertion site for target DNA-primed reverse transcription (TPRT of the inserted intron RNA. Host factors complete the insertion of the intron. By contrast, the major retrohoming pathway of introns with IEPs naturally lacking endonuclease activity, like the Sinorhizobium meliloti intron RmInt1, is thought to involve insertion of the intron RNA into the template for lagging strand DNA synthesis ahead of the replication fork, with possible use of the nascent strand to prime reverse transcription of the intron RNA. The host factors influencing the retrohoming pathway of such introns have not yet been described. Here, we identify key candidates likely to be involved in early and late steps of RmInt1 retrohoming. Some of these host factors are common to En+ group II intron retrohoming, but some have different functions. Our results also suggest that the retrohoming process of RmInt1 may be less dependent on the intracellular free Mg2+ concentration than those of other group II introns.

  9. Structural basis of PAM-dependent target DNA recognition by the Cas9 endonuclease.

    Science.gov (United States)

    Anders, Carolin; Niewoehner, Ole; Duerst, Alessia; Jinek, Martin

    2014-09-25

    The CRISPR-associated protein Cas9 is an RNA-guided endonuclease that cleaves double-stranded DNA bearing sequences complementary to a 20-nucleotide segment in the guide RNA. Cas9 has emerged as a versatile molecular tool for genome editing and gene expression control. RNA-guided DNA recognition and cleavage strictly require the presence of a protospacer adjacent motif (PAM) in the target DNA. Here we report a crystal structure of Streptococcus pyogenes Cas9 in complex with a single-molecule guide RNA and a target DNA containing a canonical 5'-NGG-3' PAM. The structure reveals that the PAM motif resides in a base-paired DNA duplex. The non-complementary strand GG dinucleotide is read out via major-groove interactions with conserved arginine residues from the carboxy-terminal domain of Cas9. Interactions with the minor groove of the PAM duplex and the phosphodiester group at the +1 position in the target DNA strand contribute to local strand separation immediately upstream of the PAM. These observations suggest a mechanism for PAM-dependent target DNA melting and RNA-DNA hybrid formation. Furthermore, this study establishes a framework for the rational engineering of Cas9 enzymes with novel PAM specificities.

  10. Recurrent invasion and extinction of a selfish gene.

    Science.gov (United States)

    Goddard, M R; Burt, A

    1999-11-23

    Homing endonuclease genes show super-Mendelian inheritance, which allows them to spread in populations even when they are of no benefit to the host organism. To test the idea that regular horizontal transmission is necessary for the long-term persistence of these genes, we surveyed 20 species of yeasts for the omega-homing endonuclease gene and associated group I intron. The status of omega could be categorized into three states (functional, nonfunctional, or absent), and status was not clustered on the host phylogeny. Moreover, the phylogeny of omega differed significantly from that of the host, strong evidence of horizontal transmission. Further analyses indicate that horizontal transmission is more common than transposition, and that it occurs preferentially between closely related species. Parsimony analysis and coalescent theory suggest that there have been 15 horizontal transmission events in the ancestry of our yeast species, through simulations indicate that this value is probably an underestimate. Overall, the data support a cyclical model of invasion, degeneration, and loss, followed by reinvasion, and each of these transitions is estimated to occur about once every 2 million years. The data are thus consistent with the idea that frequent horizontal transmission is necessary for the long-term persistence of homing endonuclease genes, and further, that this requirement limits these genes to organisms with easily accessible germ lines. The data also show that mitochondrial DNA sequences are transferred intact between yeast species; if other genes do not show such high levels of horizontal transmission, it would be due to lack of selection, rather than lack of opportunity.

  11. Interaction of the E. coli DNA G:T-mismatch endonuclease (vsr protein) with oligonucleotides containing its target sequence.

    Science.gov (United States)

    Turner, D P; Connolly, B A

    2000-12-15

    The Escherichia coli vsr endonuclease recognises G:T base-pair mismatches in double-stranded DNA and initiates a repair pathway by hydrolysing the phosphate group 5' to the incorrectly paired T. The enzyme shows a preference for G:T mismatches within a particular sequence context, derived from the recognition site of the E. coli dcm DNA-methyltransferase (CC[A/T]GG). Thus, the preferred substrate for the vsr protein is (CT[A/T]GG), where the underlined T is opposed by a dG base. This paper provides quantitative data for the interaction of the vsr protein with a number of oligonucleotides containing G:T mismatches. Evaluation of specificity constant (k(st)/K(D); k(st)=rate constant for single turnover, K(D)=equilibrium dissociation constant) confirms vsr's preference for a G:T mismatch within a hemi-methylated dcm sequence, i.e. the best substrate is a duplex (both strands written in the 5'-3' orientation) composed of CT[A/T]GG and C(5Me)C[T/A]GG. Conversion of the mispaired T (underlined) to dU or the d(5Me)C to dC gave poorer substrates. No interaction was observed with oligonucleotides that lacked a G:T mismatch or did not possess a dcm sequence. An analysis of the fraction of active protein, by "reverse-titration" (i.e. adding increasing amounts of DNA to a fixed amount of protein followed by gel-mobility shift analysis) showed that less than 1% of the vsr endonuclease was able to bind to the substrate. This was confirmed using "competitive titrations" (where competitor oligonucleotides are used to displace a (32)P-labelled nucleic acid from the vsr protein) and burst kinetic analysis. This result is discussed in the light of previous in vitro and in vivo data which indicate that the MutL protein may be needed for full vsr activity. Copyright 2000 Academic Press.

  12. Activation of apurinic/apyrimidinic endonuclease in human cells by reactive oxygen species and its correlation with their adaptive response to genotoxicity of free radicals

    OpenAIRE

    Ramana, Chilakamarti V.; Boldogh, Istvan; Izumi, Tadahide; Mitra, Sankar

    1998-01-01

    Apurinic/apyrimidinic (AP) endonuclease (APE; EC 4.2.99.18) plays a central role in repair of DNA damage due to reactive oxygen species (ROS) because its DNA 3′-phosphoesterase activity removes 3′ blocking groups in DNA that are generated by DNA glycosylase/AP-lyases during removal of oxidized bases and by direct ROS reaction with DNA. The major human APE (APE-1) gene is activated selectively by sublethal levels of a variety of ROS and ROS generators, including ionizing radiation, but not by ...

  13. Spin Seebeck effect and ballistic transport of quasi-acoustic magnons in room-temperature yttrium iron garnet films

    Science.gov (United States)

    Noack, Timo B.; Musiienko-Shmarova, Halyna Yu; Langner, Thomas; Heussner, Frank; Lauer, Viktor; Heinz, Björn; Bozhko, Dmytro A.; Vasyuchka, Vitaliy I.; Pomyalov, Anna; L’vov, Victor S.; Hillebrands, Burkard; Serga, Alexander A.

    2018-06-01

    We studied the transient behavior of the spin current generated by the longitudinal spin Seebeck effect (LSSE) in a set of platinum-coated yttrium iron garnet (YIG) films of different thicknesses. The LSSE was induced by means of pulsed microwave heating of the Pt layer and the spin currents were measured electrically using the inverse spin Hall effect in the same layer. We demonstrate that the time evolution of the LSSE is determined by the evolution of the thermal gradient triggering the flux of thermal magnons in the vicinity of the YIG/Pt interface. These magnons move ballistically within the YIG film with a constant group velocity, while their number decays exponentially within an effective propagation length. The ballistic flight of the magnons with energies above 20 K is a result of their almost linear dispersion law, similar to that of acoustic phonons. By fitting the time-dependent LSSE signal for different film thicknesses varying by almost an order of magnitude, we found that the effective propagation length is practically independent of the YIG film thickness. We consider this fact as strong support of a ballistic transport scenario—the ballistic propagation of quasi-acoustic magnons in room temperature YIG.

  14. Localized excitation of magnetostatic surface spin waves in yttrium iron garnet by shorted coaxial probe detected via spin pumping and rectification effect

    International Nuclear Information System (INIS)

    Soh, Wee Tee; Ong, C. K.; Peng, Bin

    2015-01-01

    We demonstrate the localized excitation and dc electrical detection of magnetostatic surface spin waves (MSSWs) in yttrium iron garnet (YIG) by a shorted coaxial probe. Thin films of NiFe and Pt are patterned at different regions onto a common bulk YIG substrate. A shorted coaxial probe is used to excite spin precession locally near various patterned regions. The dc voltages across the corresponding regions are recorded. For excitation of the Pt regions, the dc voltage spectra are dominated by the spin pumping of MSSWs from YIG, where various modes can be clearly distinguished. For the NiFe region, it is also found that spin pumping from MSSWs generated in YIG dominated the spectra, indicating that the spin pumped currents are dissipated into charge currents via the inverse Spin Hall effect (ISHE) in NiFe. For all regions, dc signals from YIG MSSWs are observed to be much stronger than the ferromagnetic resonance (FMR) uniform mode, likely due to the nature of the microwave excitation. The results indicate the potential of this probe for microwave imaging via dc detection of spin dynamics in continuous and patterned films

  15. Direct observation of magnon-phonon coupling in yttrium iron garnet

    Science.gov (United States)

    Man, Haoran; Shi, Zhong; Xu, Guangyong; Xu, Yadong; Chen, Xi; Sullivan, Sean; Zhou, Jianshi; Xia, Ke; Shi, Jing; Dai, Pengcheng

    2017-09-01

    The magnetic insulator yttrium iron garnet (YIG) with a ferrimagnetic transition temperature of ˜560 K has been widely used in microwave and spintronic devices. Anomalous features in spin Seeback effect (SSE) voltages have been observed in Pt/YIG and attributed to magnon-phonon coupling. Here, we use inelastic neutron scattering to map out low-energy spin waves and acoustic phonons of YIG at 100 K as a function of increasing magnetic field. By comparing the zero and 9.1 T data, we find that instead of splitting and opening up gaps at the spin wave and acoustic phonon dispersion intersecting points, magnon-phonon coupling in YIG enhances the hybridized scattering intensity. These results are different from expectations of conventional spin-lattice coupling, calling for different paradigms to understand the scattering process of magnon-phonon interactions and the resulting magnon polarons.

  16. Spin wave differential circuit for realization of thermally stable magnonic sensors

    Energy Technology Data Exchange (ETDEWEB)

    Goto, Taichi, E-mail: goto@ee.tut.ac.jp; Kanazawa, Naoki; Buyandalai, Altansargai; Takagi, Hiroyuki; Nakamura, Yuichi; Inoue, Mitsuteru [Department of Electrical and Electronic Information Engineering, Toyohashi University of Technology, 1-1 Hibari-Ga-Oka, Tempaku, Toyohashi, Aichi 441-8580 (Japan); Okajima, Shingo; Hasegawa, Takashi [Murata Manufacturing Co., Ltd., Kyoto 617-8555 (Japan); Granovsky, Alexander B. [Faculty of Physics, Moscow State University, Leninskie Gory, Moscow 119992 (Russian Federation); Sekiguchi, Koji [Department of Physics, Keio University, Yokohama 223-8522 (Japan); JST-PRESTO, Kawaguchi, Saitama 332-0012 (Japan); Ross, Caroline A. [Department of Materials Science and Engineering, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, Massachusetts 02139 (United States)

    2015-03-30

    A magnetic-field sensor with a high sensitivity of 38 pT/Hz was demonstrated. By utilizing a spin-wave differential circuit (SWDC) using two yttrium iron garnet (YIG) films, the temperature sensitivity was suppressed, and the thermal stability of the phase of the spin waves was −0.0095° K{sup −1}, which is three orders of magnitude better than a simple YIG-based sensor, ∼20° K{sup −1}. The SWDC architecture opens the way to design YIG-based magnonic devices.

  17. Spin wave differential circuit for realization of thermally stable magnonic sensors

    International Nuclear Information System (INIS)

    Goto, Taichi; Kanazawa, Naoki; Buyandalai, Altansargai; Takagi, Hiroyuki; Nakamura, Yuichi; Inoue, Mitsuteru; Okajima, Shingo; Hasegawa, Takashi; Granovsky, Alexander B.; Sekiguchi, Koji; Ross, Caroline A.

    2015-01-01

    A magnetic-field sensor with a high sensitivity of 38 pT/Hz was demonstrated. By utilizing a spin-wave differential circuit (SWDC) using two yttrium iron garnet (YIG) films, the temperature sensitivity was suppressed, and the thermal stability of the phase of the spin waves was −0.0095° K −1 , which is three orders of magnitude better than a simple YIG-based sensor, ∼20° K −1 . The SWDC architecture opens the way to design YIG-based magnonic devices

  18. One-tube loop-mediated isothermal amplification combined with restriction endonuclease digestion and ELISA for colorimetric detection of resistance to isoniazid, ethambutol and streptomycin in Mycobacterium tuberculosis isolates.

    Science.gov (United States)

    Lee, Mei-Feng; Chen, Yen-Hsu; Hsu, Hui-Jine; Peng, Chien-Fang

    2010-10-01

    In this study, we designed a simple and rapid colorimetric detection method, a one-tube loop-mediated isothermal amplification (LAMP)-PCR-hybridization-restriction endonuclease-ELISA [one-tube LAMP-PCR-HY-RE-ELISA] system, to detect resistance to isoniazid, ethambutol and streptomycin in strains of Mycobacterium tuberculosis isolated from clinical specimens. The clinical performance of this method for detecting isoniazid-resistant, ethambutol-resistant and streptomycin-resistant isolates of M. tuberculosis showed 98.9%, 94.3% and 93.8%, respectively. This assay is rapid and convenient that can be performed within one working day. One-tube LAMP-PCR-HY-RE-ELISA system was designed based on hot spot point mutations in target drug-resistant genes, using LAMP-PCR, hybridization, digestion with restriction endonuclease and colorimetric method of ELISA. In this study, LAMP assay was used to amplify DNA from drug-resistant M. tuberculosis, and ELISA was used for colorimetrical determination. This assay will be a useful tool for rapid diagnosis of mutant codons in strains of M. tuberculosis for isoniazid at katG 315 and katG 463, ethambutol at embB 306 and embB 497, and streptomycin at rpsL 43. Crown Copyright © 2010. Published by Elsevier B.V. All rights reserved.

  19. Structural Plasticity of PAM Recognition by Engineered Variants of the RNA-Guided Endonuclease Cas9.

    Science.gov (United States)

    Anders, Carolin; Bargsten, Katja; Jinek, Martin

    2016-03-17

    The RNA-guided endonuclease Cas9 from Streptococcus pyogenes (SpCas9) forms the core of a powerful genome editing technology. DNA cleavage by SpCas9 is dependent on the presence of a 5'-NGG-3' protospacer adjacent motif (PAM) in the target DNA, restricting the choice of targetable sequences. To address this limitation, artificial SpCas9 variants with altered PAM specificities have recently been developed. Here we report crystal structures of the VQR, EQR, and VRER SpCas9 variants bound to target DNAs containing their preferred PAM sequences. The structures reveal that the non-canonical PAMs are recognized by an induced fit mechanism. Besides mediating sequence-specific base recognition, the amino acid substitutions introduced in the SpCas9 variants facilitate conformational remodeling of the PAM region of the bound DNA. Guided by the structural data, we engineered a SpCas9 variant that specifically recognizes NAAG PAMs. Taken together, these studies inform further development of Cas9-based genome editing tools. Copyright © 2016 Elsevier Inc. All rights reserved.

  20. How quantum entanglement in DNA synchronizes double-strand breakage by type II restriction endonucleases.

    Science.gov (United States)

    Kurian, P; Dunston, G; Lindesay, J

    2016-02-21

    Macroscopic quantum effects in living systems have been studied widely in pursuit of fundamental explanations for biological energy transport and sensing. While it is known that type II endonucleases, the largest class of restriction enzymes, induce DNA double-strand breaks by attacking phosphodiester bonds, the mechanism by which simultaneous cutting is coordinated between the catalytic centers remains unclear. We propose a quantum mechanical model for collective electronic behavior in the DNA helix, where dipole-dipole oscillations are quantized through boundary conditions imposed by the enzyme. Zero-point modes of coherent oscillations would provide the energy required for double-strand breakage. Such quanta may be preserved in the presence of thermal noise by the enzyme's displacement of water surrounding the DNA recognition sequence. The enzyme thus serves as a decoherence shield. Palindromic mirror symmetry of the enzyme-DNA complex should conserve parity, because symmetric bond-breaking ceases when the symmetry of the complex is violated or when physiological parameters are perturbed from optima. Persistent correlations in DNA across longer spatial separations-a possible signature of quantum entanglement-may be explained by such a mechanism. Copyright © 2015 Elsevier Ltd. All rights reserved.

  1. Molecular characterization of a new member of the lariat capping twin-ribozyme introns

    DEFF Research Database (Denmark)

    Tang, Yunjia; Nielsen, Henrik; Masquida, Benoît

    2014-01-01

    BACKGROUND: Twin-ribozyme introns represent a complex class of mobile group I introns that harbour a lariat capping (LC) ribozyme and a homing endonuclease gene embedded in a conventional self-splicing group I ribozyme (GIR2). Twin-ribozyme introns have so far been confined to nucleolar DNA in Na...

  2. Modulation of the Pyrococcus abyssi NucS endonuclease activity by replication clamp at functional and structural levels.

    Science.gov (United States)

    Creze, Christophe; Ligabue, Alessio; Laurent, Sébastien; Lestini, Roxane; Laptenok, Sergey P; Khun, Joelle; Vos, Marten H; Czjzek, Mirjam; Myllykallio, Hannu; Flament, Didier

    2012-05-04

    Pyrococcus abyssi NucS is the founding member of a new family of structure-specific DNA endonucleases that interact with the replication clamp proliferating cell nuclear antigen (PCNA). Using a combination of small angle x-ray scattering and surface plasmon resonance analyses, we demonstrate the formation of a stable complex in solution, in which one molecule of the PabNucS homodimer binds to the outside surface of the PabPCNA homotrimer. Using fluorescent labels, PCNA is shown to increase the binding affinity of NucS toward single-strand/double-strand junctions on 5' and 3' flaps, as well as to modulate the cleavage specificity on the branched DNA structures. Our results indicate that the presence of a single major contact between the PabNucS and PabPCNA proteins, together with the complex-induced DNA bending, facilitate conformational flexibility required for specific cleavage at the single-strand/double-strand DNA junction.

  3. Modulation of the Pyrococcus abyssi NucS Endonuclease Activity by Replication Clamp at Functional and Structural Levels*

    Science.gov (United States)

    Creze, Christophe; Ligabue, Alessio; Laurent, Sébastien; Lestini, Roxane; Laptenok, Sergey P.; Khun, Joelle; Vos, Marten H.; Czjzek, Mirjam; Myllykallio, Hannu; Flament, Didier

    2012-01-01

    Pyrococcus abyssi NucS is the founding member of a new family of structure-specific DNA endonucleases that interact with the replication clamp proliferating cell nuclear antigen (PCNA). Using a combination of small angle x-ray scattering and surface plasmon resonance analyses, we demonstrate the formation of a stable complex in solution, in which one molecule of the PabNucS homodimer binds to the outside surface of the PabPCNA homotrimer. Using fluorescent labels, PCNA is shown to increase the binding affinity of NucS toward single-strand/double-strand junctions on 5′ and 3′ flaps, as well as to modulate the cleavage specificity on the branched DNA structures. Our results indicate that the presence of a single major contact between the PabNucS and PabPCNA proteins, together with the complex-induced DNA bending, facilitate conformational flexibility required for specific cleavage at the single-strand/double-strand DNA junction. PMID:22431731

  4. [Effect of endonuclease G depletion on plasmid DNA uptake and levels of homologous recombination in hela cells].

    Science.gov (United States)

    Misic, V; El-Mogy, M; Geng, S; Haj-Ahmad, Y

    2016-01-01

    Endonuclease G (EndoG) is a mitochondrial apoptosis regulator that also has roles outside of programmed cell death. It has been implicated as a defence DNase involved in the degradation of exogenous DNA after transfection of mammalian cells and in homologous recombination of viral and endogenous DNA. In this study, we looked at the effect of EndoG depletion on plasmid DNA uptake and the levels of homologous recombination in HeLa cells. We show that the proposed defence role of EndoG against uptake of non-viral DNA vectors does not extend to the cervical carcinoma HeLa cells, as targeting of EndoG expression by RNA interference failed to increase intracellular plasmid DNA levels. However, reducing EndoG levels in HeLa cells resulted in a statistically significant reduction of homologous recombination between two plasmid DNA substrates. These findings suggest that non-viral DNA vectors are also substrates for EndoG in its role in homologous recombination.

  5. Multipronged regulatory functions of a novel endonuclease (TieA) from Helicobacter pylori.

    Science.gov (United States)

    Devi, Savita; Ansari, Suhail A; Tenguria, Shivendra; Kumar, Naveen; Ahmed, Niyaz

    2016-11-02

    Helicobacter pylori portrays a classical paradigm of persistent bacterial infections. A well balanced homeostasis of bacterial effector functions and host responses is purported to be the key in achieving long term colonization in specific hosts. H. pylori nucleases have been shown to assist in natural transformation, but their role in virulence and colonization remains elusive. Therefore, it is imperative to understand the involvement of these nucleases in the pathogenesis of H. pylori Here, we report the multifaceted role of a TNFR-1 interacting endonuclease A (TieA) from H. pylori. tieA expression is differentially regulated in response to environmental stress and post adherence to gastric epithelial cells. Studies with isogenic knockouts of tieA revealed it to be a secretory protein which translocates into the host gastric epithelial cells independent of a type IV secretion system, gets phosphorylated by DNA-PK kinase and auto-phosphorylates as serine kinase. Furthermore, TieA binds to and cleaves DNA in a non-specific manner and promotes Fas mediated apoptosis in AGS cells. Additionally, TieA induced pro-inflammatory cytokine secretion via activation of transcription factor AP-1 and signaled through MAP kinase pathway. Collectively, TieA with its multipronged and moonlighting functions could facilitate H. pylori in maintaining a balance of bacterial adaptation, and elimination by the host responses. © The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.

  6. Observation of the Spin Peltier Effect for Magnetic Insulators

    NARCIS (Netherlands)

    Flipse, J.; Dejene, F.K.; Wagenaar, D.; Bauer, G.E.W.; Ben Youssef, J.; Van Wees, B.J.

    2014-01-01

    We report the observation of the spin Peltier effect (SPE) in the ferrimagnetic insulator yttrium iron garnet (YIG), i.e., a heat current generated by a spin current flowing through a platinum (Pt)|YIG interface. The effect can be explained by the spin transfer torque that transforms the spin

  7. Sequencing by ligation variation with endonuclease V digestion and deoxyinosine-containing query oligonucleotides

    Directory of Open Access Journals (Sweden)

    Ho Antoine

    2011-12-01

    Full Text Available Abstract Background Sequencing-by-ligation (SBL is one of several next-generation sequencing methods that has been developed for massive sequencing of DNA immobilized on arrayed beads (or other clonal amplicons. SBL has the advantage of being easy to implement and accessible to all because it can be performed with off-the-shelf reagents. However, SBL has the limitation of very short read lengths. Results To overcome the read length limitation, research groups have developed complex library preparation processes, which can be time-consuming, difficult, and result in low complexity libraries. Herein we describe a variation on traditional SBL protocols that extends the number of sequential bases that can be sequenced by using Endonuclease V to nick a query primer, thus leaving a ligatable end extended into the unknown sequence for further SBL cycles. To demonstrate the protocol, we constructed a known DNA sequence and utilized our SBL variation, cyclic SBL (cSBL, to resequence this region. Using our method, we were able to read thirteen contiguous bases in the 3' - 5' direction. Conclusions Combining this read length with sequencing in the 5' - 3' direction would allow a read length of over twenty bases on a single tage. Implementing mate-paired tags and this SBL variation could enable > 95% coverage of the genome.

  8. Efficient genome editing in hematopoietic stem cells with helper-dependent Ad5/35 vectors expressing site-specific endonucleases under microRNA regulation

    Directory of Open Access Journals (Sweden)

    Kamola Saydaminova

    Full Text Available Genome editing with site-specific endonucleases has implications for basic biomedical research as well as for gene therapy. We generated helper-dependent, capsid-modified adenovirus (HD-Ad5/35 vectors for zinc-finger nuclease (ZFN– or transcription activator-like effector nuclease (TALEN–mediated genome editing in human CD34+ hematopoietic stem cells (HSCs from mobilized adult donors. The production of these vectors required that ZFN and TALEN expression in HD-Ad5/35 producer 293-Cre cells was suppressed. To do this, we developed a microRNA (miRNA-based system for regulation of gene expression based on miRNA expression profiling of 293-Cre and CD34+ cells. Using miR-183-5p and miR-218-5p based regulation of transgene gene expression, we first produced an HD-Ad5/35 vector expressing a ZFN specific to the HIV coreceptor gene ccr5. We demonstrated that HD-Ad5/35.ZFNmiR vector conferred ccr5 knock out in primitive HSC (i.e., long-term culture initiating cells and NOD/SCID repopulating cells. The ccr5 gene disruption frequency achieved in engrafted HSCs found in the bone marrow of transplanted mice is clinically relevant for HIV therapy considering that these cells can give rise to multiple lineages, including all the lineages that represent targets and reservoirs for HIV. We produced a second HD-Ad5/35 vector expressing a TALEN targeting the DNase hypersensitivity region 2 (HS2 within the globin locus control region. This vector has potential for targeted gene correction in hemoglobinopathies. The miRNA regulated HD-Ad5/35 vector platform for expression of site-specific endonucleases has numerous advantages over currently used vectors as a tool for genome engineering of HSCs for therapeutic purposes.

  9. PAM-Dependent Target DNA Recognition and Cleavage by C2c1 CRISPR-Cas Endonuclease

    Energy Technology Data Exchange (ETDEWEB)

    Yang, Hui; Gao, Pu; Rajashankar, Kanagalaghatta R.; Patel, Dinshaw J. (MSKCC); (Cornell); (Chinese Aca. Sci.)

    2016-12-01

    C2c1 is a newly identified guide RNA-mediated type V-B CRISPR-Cas endonuclease that site-specifically targets and cleaves both strands of target DNA. We have determined crystal structures of Alicyclobacillus acidoterrestris C2c1 (AacC2c1) bound to sgRNA as a binary complex and to target DNAs as ternary complexes, thereby capturing catalytically competent conformations of AacC2c1 with both target and non-target DNA strands independently positioned within a single RuvC catalytic pocket. Moreover, C2c1-mediated cleavage results in a staggered seven-nucleotide break of target DNA. crRNA adopts a pre-ordered five-nucleotide A-form seed sequence in the binary complex, with release of an inserted tryptophan, facilitating zippering up of 20-bp guide RNA:target DNA heteroduplex on ternary complex formation. Notably, the PAM-interacting cleft adopts a “locked” conformation on ternary complex formation. Structural comparison of C2c1 ternary complexes with their Cas9 and Cpf1 counterparts highlights the diverse mechanisms adopted by these distinct CRISPR-Cas systems, thereby broadening and enhancing their applicability as genome editing tools.

  10. Magnetophotonic crystals based on yttrium-iron-garnet infiltrated opals: Magnetization-induced second-harmonic generation

    Science.gov (United States)

    Murzina, T. V.; Kim, E. M.; Kapra, R. V.; Moshnina, I. V.; Aktsipetrov, O. A.; Kurdyukov, D. A.; Kaplan, S. F.; Golubev, V. G.; Bader, M. A.; Marowsky, G.

    2006-01-01

    Three-dimensional magnetophotonic crystals (MPCs) based on artificial opals infiltrated by yttrium iron garnet (YIG) are fabricated and their structural, optical, and nonlinear optical properties are studied. The formation of the crystalline YIG inside the opal matrix is checked by x-ray analysis. Two templates are used for the infiltration by YIG: bare opals and those covered by a thin platinum film. Optical second-harmonic generation (SHG) technique is used to study the magnetization-induced nonlinear-optical properties of the composed MPCs. A high nonlinear magneto-optical Kerr effect in the SHG intensity is observed at the edge of the photonic band gap of the MPCs.

  11. Total sequence decomposition distinguishes functional modules, "molegos" in apurinic/apyrimidinic endonucleases

    Directory of Open Access Journals (Sweden)

    Braun Werner

    2002-11-01

    Full Text Available Abstract Background Total sequence decomposition, using the web-based MASIA tool, identifies areas of conservation in aligned protein sequences. By structurally annotating these motifs, the sequence can be parsed into individual building blocks, molecular legos ("molegos", that can eventually be related to function. Here, the approach is applied to the apurinic/apyrimidinic endonuclease (APE DNA repair proteins, essential enzymes that have been highly conserved throughout evolution. The APEs, DNase-1 and inositol 5'-polyphosphate phosphatases (IPP form a superfamily that catalyze metal ion based phosphorolysis, but recognize different substrates. Results MASIA decomposition of APE yielded 12 sequence motifs, 10 of which are also structurally conserved within the family and are designated as molegos. The 12 motifs include all the residues known to be essential for DNA cleavage by APE. Five of these molegos are sequentially and structurally conserved in DNase-1 and the IPP family. Correcting the sequence alignment to match the residues at the ends of two of the molegos that are absolutely conserved in each of the three families greatly improved the local structural alignment of APEs, DNase-1 and synaptojanin. Comparing substrate/product binding of molegos common to DNase-1 showed that those distinctive for APEs are not directly involved in cleavage, but establish protein-DNA interactions 3' to the abasic site. These additional bonds enhance both specific binding to damaged DNA and the processivity of APE1. Conclusion A modular approach can improve structurally predictive alignments of homologous proteins with low sequence identity and reveal residues peripheral to the traditional "active site" that control the specificity of enzymatic activity.

  12. Electric field tuning of magnetism in heterostructure of yttrium iron garnet film/lead magnesium niobate-lead zirconate titanate ceramic

    Science.gov (United States)

    Lian, Jianyun; Ponchel, Freddy; Tiercelin, Nicolas; Chen, Ying; Rémiens, Denis; Lasri, Tuami; Wang, Genshui; Pernod, Philippe; Zhang, Wenbin; Dong, Xianlin

    2018-04-01

    In this paper, the converse magnetoelectric (CME) effect by electric field tuning of magnetization in an original heterostructure composed of a polycrystalline yttrium iron garnet (YIG) film and a lead magnesium niobate-lead zirconate titanate (PMN-PZT) ceramic is presented. The magnetic performances of the YIG films with different thicknesses under a DC electric field applied to the PMN-PZT ceramics and a bias magnetic field are investigated. All the magnetization-electric field curves are found to be in good agreement with the butterfly like strain curve of the PMN-PZT ceramic. Both the sharp deformation of about 2.5‰ of PMN-PZT and the easy magnetization switching of YIG are proposed to be the reasons for the strongest CME interaction in the composite at the small electric coercive field of PMN-PZT (4.1 kV/cm) and the small magnetic coercive field of YIG (20 Oe) where the magnetic susceptibility reaches its maximum value. A remarkable CME coefficient of 3.1 × 10-7 s/m is obtained in the system with a 600 nm-thick YIG film. This heterostructure combining multiferroics and partially magnetized ferrite concepts is able to operate under a small or even in the absence of an external bias magnetic field and is more compact and power efficient than the traditional magnetoelectric devices.

  13. Preparing magnetic yttrium iron garnet nanodot arrays by ultrathin anodic alumina template on silicon substrate

    Energy Technology Data Exchange (ETDEWEB)

    Zheng, Hui; Han, Mangui, E-mail: han-mangui@yahoo.com; Deng, Longjiang [National Engineering Research Center of Electromagnetic Radiation Control Materials, University of Electronic Science and Technology of China, Chengdu 610054 (China); Zheng, Liang; Zheng, Peng; Qin, Huibin [Institute of Electron Device and Application, Hangzhou Dianzi University, Hangzhou 310008 (China); Wu, Qiong [Magnetism Key Laboratory of Zhejiang Province, China Jiliang University, Hangzhou 310018 (China)

    2015-08-10

    Ultrahigh density periodically ordered magnetic yttrium iron garnet (Y{sub 3}Fe{sub 5}O{sub 12}, YIG) nanodot arrays have been prepared by pulsed laser deposition through an ultrathin alumina mask (UTAM). UTAM having periodically ordered circularly shaped holes with 350 nm in diameter, 450 nm in inter-pore distance, and 700 nm in height has been prepared on silicon substrate. Furthermore, the microstructure and magnetic properties of YIG nanodot arrays have been characterized. Nanodot arrays with a sharp distribution in diameter centered at 340 nm with standard deviation of 10 nm have been fabricated. Moreover, typical hysteresis loops and ferromagnetic resonance spectra in in-plane and out-of-plane revealed that this unique structure greatly influences the magnetics properties of YIG. First, coercivity of YIG nanodot arrays in in-plane was increased about from 15 Oe of YIG films to 500 Oe. Then, the degree of uniformity about nanodot height decided that two or more resonance peaks in out-of-plane were detected in the spectra. The peak-to-peak linewidth values were about 94 Oe and 40 Oe in the parallel and perpendicular directions, respectively, which indicated that the values were larger by the two-magnon scattering. Consequently, this pattering method creates opportunities for studying physics in oxide nanomagnets and may be applied in spin-wave devices.

  14. Location, Location, Location: Characteristics and Services of Long-Stay Home Care Recipients in Retirement Homes Compared to Others in Private Homes and Long-Term Care Homes.

    Science.gov (United States)

    Poss, Jeffrey W; Sinn, Chi-Ling Joanna; Grinchenko, Galina; Blums, Jane; Peirce, Tom; Hirdes, John

    2017-02-01

    We examine recipients of publicly funded ongoing care in a single Ontario jurisdiction who reside in three different settings: long-stay home care patients in private homes and apartments, other patients in retirement homes and residents of long-term care homes, using interRAI assessment instruments. Among home care patients, those in retirement homes have higher proportions of dementia and moderate cognitive impairment, less supportive informal care systems as well as more personal care and nursing services above those provided by the public home care system, more frequent but shorter home support visits and lower than expected public home care expenditures. These lower expenditures may be because of efficiency of care delivery or by retirement homes providing some services otherwise provided by the public home care system. Although persons in each setting are mostly older adults with high degrees of frailty and medical complexity, long-term care home residents show distinctly higher needs. We estimate that 40% of retirement home residents are long-stay home care patients, and they comprise about one in six of this Community Care Access Centre's long-stay patients. Copyright © 2017 Longwoods Publishing.

  15. Construction of an easy-to-use CRISPR-Cas9 system by patching a newly designed EXIT circuit.

    Science.gov (United States)

    Tang, Qiang; Lou, Chunbo; Liu, Shuang-Jiang

    2017-01-01

    Plasmid-borne genetic editing tools, including the widely used CRISPR-Cas9 system, have greatly facilitated bacterial programming to obtain novel functionalities. However, the lack of effective post-editing plasmid elimination methods impedes follow-up genetic manipulation or application. Conventional strategies including exposure to physical and chemical treatments, or exploiting temperature-sensitive replication origins have several drawbacks (e.g., they are limited for efficiency and are time-consuming). Therefore, the demand is apparent for easy and rapid elimination of the tool plasmids from their bacterial hosts after genetic manipulation. To bridge this gap, we designed a novel EXIT circuit with the homing endonuclease, which can be exploited for rapid and efficient elimination of various plasmids with diverse replication origins. As a proof of concept, we validated the EXIT circuit in Escherichia coli by harnessing homing endonuclease I- Sce I and its cleavage site. When integrated into multiple plasmids with different origins, the EXIT circuit allowed them to be eliminated from the host cells, simultaneously. By combining the widely used plasmid-borne CRISPR-Cas9 system and the EXIT circuit, we constructed an easy-to-use CRISPR-Cas9 system that eliminated the Cas9- and the single-guide RNA (sgRNA)-encoding plasmids in one-step. Within 3 days, we successfully constructed an atrazine-degrading E. coli strain, thus further demonstrating the advantage of this new CRISPR-Cas9 system for bacterial genome editing. Our novel EXIT circuit, which exploits the homing endonuclease I- Sce I, enables plasmid(s) with different replication origins to be eliminated from their host cells rapidly and efficiently. We also developed an easy-to-use CRISPR-Cas9 system with the EXIT circuit, and this new system can be widely applied to bacterial genome editing.

  16. Homing oneself

    DEFF Research Database (Denmark)

    Winther, Ida Wentzel

    2009-01-01

    What is home? A building, a physical and mental phenomenon, or a concept?  There are many homes and ways `to home oneself´. Many of us quite often dwell in other places than at home (as professional commuters between two places, as travellers staying in hotels, as children of divorced parents...

  17. Aromatic residues located close to the active center are essential for the catalytic reaction of flap endonuclease-1 from hyperthermophilic archaeon Pyrococcus horikoshii.

    Science.gov (United States)

    Matsui, Eriko; Abe, Junko; Yokoyama, Hideshi; Matsui, Ikuo

    2004-04-16

    Flap endonuclease-1 (FEN-1) possessing 5'-flap endonuclease and 5'-->3' exonuclease activity plays important roles in DNA replication and repair. In this study, the kinetic parameters of mutants at highly conserved aromatic residues, Tyr33, Phe35, Phe79, and Phe278-Phe279, in the vicinity of the catalytic centers of FEN-1 were examined. The substitution of these aromatic residues with alanine led to a large reduction in kcat values, although these mutants retained Km values similar to that of the wild-type enzyme. Notably, the kcat of Y33A and F79A decreased 333-fold and 71-fold, respectively, compared with that of the wild-type enzyme. The aromatic residues Tyr33 and Phe79, and the aromatic cluster Phe278-Phe279 mainly contributed to the recognition of the substrates without the 3' projection of the upstream strand (the nick, 5'-recess-end, single-flap, and pseudo-Y substrates) for the both exo- and endo-activities, but played minor roles in recognizing the substrates with the 3' projection (the double flap substrate and the nick substrate with the 3' projection). The replacement of Tyr33, Phe79, and Phe278-Phe279, with non-charged aromatic residues, but not with aliphatic hydrophobic residues, recovered the kcat values almost fully for the substrates without the 3' projection of the upstream strand, suggesting that the aromatic groups of Tyr33, Phe79, and Phe278-Phe279 might be involved in the catalytic reaction, probably via multiple stacking interactions with nucleotide bases. The stacking interactions of Tyr33 and Phe79 might play important roles in fixing the template strand and the downstream strand, respectively, in close proximity to the active center to achieve the productive transient state leading to the hydrolysis.

  18. Family and home characteristics correlate with mold in homes.

    Science.gov (United States)

    Reponen, Tiina; Levin, Linda; Zheng, Shu; Vesper, Stephen; Ryan, Patrick; Grinshpun, Sergey A; LeMasters, Grace

    2013-07-01

    Previously, we demonstrated that infants residing in homes with higher Environmental Relative Moldiness Index were at greater risk for developing asthma by age seven. The purpose of this analysis was to identify the family and home characteristics associated with higher moldiness index values in infants' homes at age one. Univariate linear regression of each characteristic determined that family factors associated with moldiness index were race and income. Home characteristics associated with the moldiness index values were: air conditioning, carpet, age of the home, season of home assessment, and house dust mite allergen. Parental history of asthma, use of dehumidifier, visible mold, dog and cat allergen levels were not associated with moldiness index. Results of multiple linear regression showed that older homes had 2.9 units higher moldiness index (95% confidence interval [CI]=0.4, 5.4), whereas homes with central air conditioning had 2.5 units lower moldiness index (95% CI=-4.7, -0.4). In addition, higher dust mite allergen levels and carpeting were positively and negatively associated with higher moldiness index, respectively. Because older homes and lack of air conditioning were also correlated with race and lower income, whereas carpeting was associated with newer homes, the multivariate analyses suggests that lower overall socioeconomic position is associated with higher moldiness index values. This may lead to increased asthma risk in homes inhabited by susceptible, vulnerable population subgroups. Further, age of the home was a surrogate of income, race and carpeting in our population; thus the use of these factors should carefully be evaluated in future studies. Copyright © 2013 Elsevier Inc. All rights reserved.

  19. Single-nitrogen-vacancy-center quantum memory for a superconducting flux qubit mediated by a ferromagnet

    Science.gov (United States)

    Lai, Yen-Yu; Lin, Guin-Dar; Twamley, Jason; Goan, Hsi-Sheng

    2018-05-01

    We propose a quantum memory scheme to transfer and store the quantum state of a superconducting flux qubit (FQ) into the electron spin of a single nitrogen-vacancy (NV) center in diamond via yttrium iron garnet (YIG), a ferromagnet. Unlike an ensemble of NV centers, the YIG moderator can enhance the effective FQ-NV-center coupling strength without introducing additional appreciable decoherence. We derive the effective interaction between the FQ and the NV center by tracing out the degrees of freedom of the collective mode of the YIG spins. We demonstrate the transfer, storage, and retrieval procedures, taking into account the effects of spontaneous decay and pure dephasing. Using realistic experimental parameters for the FQ, NV center and YIG, we find that a combined transfer, storage, and retrieval fidelity higher than 0.9, with a long storage time of 10 ms, can be achieved. This hybrid system not only acts as a promising quantum memory, but also provides an example of enhanced coupling between various systems through collective degrees of freedom.

  20. New ferrimagnetic biocomposite film based in collagen and yttrium iron garnet

    Directory of Open Access Journals (Sweden)

    2010-12-01

    Full Text Available In recent years a great interest in the study of the association of magnetic with biological material for bioapplications has been observed in the literature. This work analyses the development of new magnetic biocomposite films from a magnetic ferrite and a biopolymer. Magnetic and dielectric properties of Y3Fe5O12 (YIG/collagen composite films were studied as a function of the YIG concentration. This biocomposite was also characterized by Infrared Spectroscopy (IR, Thermal Analysis (DSC and TG and scanning electron microspcopic (SEM methods. The magnetization and dielectric measurements were performed at room temperature. The results demonstrated that ferrimagnetic garnet (YIG and collagen (Col can be used to obtain a homogeneous composite. All the composite films showed a ferromagnetic behavior and they were characterized as a soft magnet material. These results show that Col-YIG biocomposites are biological films with magnetic properties that can be employed as a versatile performance materials, due to their flexible dielectric and magnetic features. They could be used as electronic devices in biological applications.

  1. Identification of four families of yCCR4- and Mg2+-dependent endonuclease-related proteins in higher eukaryotes, and characterization of orthologs of yCCR4 with a conserved leucine-rich repeat essential for hCAF1/hPOP2 binding

    Directory of Open Access Journals (Sweden)

    Corbo Laura

    2001-11-01

    Full Text Available Abstract Background The yeast yCCR4 factor belongs to the CCR4-NOT transcriptional regulatory complex, in which it interacts, through its leucine-rich repeat (LRR motif with yPOP2. Recently, yCCR4 was shown to be a component of the major cytoplasmic mRNA deadenylase complex, and to contain a fold related to the Mg2+-dependent endonuclease core. Results Here, we report the identification of nineteen yCCR4-related proteins in eukaryotes (including yeast, plants and animals, which all contain the yCCR4 endonuclease-like fold, with highly conserved CCR4-specific residues. Phylogenetic and genomic analyses show that they form four distinct families, one of which contains the yCCR4 orthologs. The orthologs in animals possess a leucine-rich repeat domain. We show, using two-hybrid and far-Western assays, that the human member binds to the human yPOP2 homologs, i.e. hCAF1 and hPOP2, in a LRR-dependent manner. Conclusions We have identified the mammalian orthologs of yCCR4 and have shown that the human member binds to the human yPOP2 homologs, thus strongly suggesting conservation of the CCR4-NOT complex from yeast to human. All members of the four identified yCCR4-related protein families show stricking conservation of the endonuclease-like catalytic motifs of the yCCR4 C-terminal domain and therefore constitute a new family of potential deadenylases in mammals.

  2. The elastic network model reveals a consistent picture on intrinsic functional dynamics of type II restriction endonucleases

    International Nuclear Information System (INIS)

    Uyar, A; Kurkcuoglu, O; Doruker, P; Nilsson, L

    2011-01-01

    The vibrational dynamics of various type II restriction endonucleases, in complex with cognate/non-cognate DNA and in the apo form, are investigated with the elastic network model in order to reveal common functional mechanisms in this enzyme family. Scissor-like and tong-like motions observed in the slowest modes of all enzymes and their complexes point to common DNA recognition and cleavage mechanisms. Normal mode analysis further points out that the scissor-like motion has an important role in differentiating between cognate and non-cognate sequences at the recognition site, thus implying its catalytic relevance. Flexible regions observed around the DNA-binding site of the enzyme usually concentrate on the highly conserved β-strands, especially after DNA binding. These β-strands may have a structurally stabilizing role in functional dynamics for target site recognition and cleavage. In addition, hot spot residues based on high-frequency modes reveal possible communication pathways between the two distant cleavage sites in the enzyme family. Some of these hot spots also exist on the shortest path between the catalytic sites and are highly conserved

  3. Phosphate steering by Flap Endonuclease 1 promotes 5′-flap specificity and incision to prevent genome instability

    KAUST Repository

    Tsutakawa, Susan E.

    2017-06-27

    DNA replication and repair enzyme Flap Endonuclease 1 (FEN1) is vital for genome integrity, and FEN1 mutations arise in multiple cancers. FEN1 precisely cleaves single-stranded (ss) 5\\'-flaps one nucleotide into duplex (ds) DNA. Yet, how FEN1 selects for but does not incise the ss 5\\'-flap was enigmatic. Here we combine crystallographic, biochemical and genetic analyses to show that two dsDNA binding sites set the 5\\'polarity and to reveal unexpected control of the DNA phosphodiester backbone by electrostatic interactions. Via phosphate steering\\', basic residues energetically steer an inverted ss 5\\'-flap through a gateway over FEN1\\'s active site and shift dsDNA for catalysis. Mutations of these residues cause an 18,000-fold reduction in catalytic rate in vitro and large-scale trinucleotide (GAA) repeat expansions in vivo, implying failed phosphate-steering promotes an unanticipated lagging-strand template-switch mechanism during replication. Thus, phosphate steering is an unappreciated FEN1 function that enforces 5\\'-flap specificity and catalysis, preventing genomic instability.

  4. 24 CFR 982.612 - Group home: State approval of group home.

    Science.gov (United States)

    2010-04-01

    ... 24 Housing and Urban Development 4 2010-04-01 2010-04-01 false Group home: State approval of group... Types Group Home § 982.612 Group home: State approval of group home. A group home must be licensed..., Retardation, or Social Services) as a group home for elderly persons or persons with disabilities. ...

  5. ThinkHome Energy Efficiency in Future Smart Homes

    OpenAIRE

    Christian Reinisch; Mario J. Kofler; Félix Iglesias; Wolfgang Kastner

    2011-01-01

    Abstract Smart homes have been viewed with increasing interest by both home owners and the research community in the past few years. One reason for this development is that the use of modern automation technology in the home or building promises considerable savings of energy, therefore, simultaneously reducing the operational costs of the building over its whole lifecycle. However, the full potential of smart homes still lies fallow, due to the complexity and diversity of the systems, badly ...

  6. Human AP Endonuclease 1: A Potential Marker for the Prediction of Environmental Carcinogenesis Risk

    Directory of Open Access Journals (Sweden)

    Jae Sung Park

    2014-01-01

    Full Text Available Human apurinic/apyrimidinic endonuclease 1 (APE1 functions mainly in DNA repair as an enzyme removing AP sites and in redox signaling as a coactivator of various transcription factors. Based on these multifunctions of APE1 within cells, numerous studies have reported that the alteration of APE1 could be a crucial factor in development of human diseases such as cancer and neurodegeneration. In fact, the study on the combination of an individual’s genetic make-up with environmental factors (gene-environment interaction is of great importance to understand the development of diseases, especially lethal diseases including cancer. Recent reports have suggested that the human carcinogenic risk following exposure to environmental toxicants is affected by APE1 alterations in terms of gene-environment interactions. In this review, we initially outline the critical APE1 functions in the various intracellular mechanisms including DNA repair and redox regulation and its roles in human diseases. Several findings demonstrate that the change in expression and activity as well as genetic variability of APE1 caused by environmental chemical (e.g., heavy metals and cigarette smoke and physical carcinogens (ultraviolet and ionizing radiation is likely associated with various cancers. These enable us to ultimately suggest APE1 as a vital marker for the prediction of environmental carcinogenesis risk.

  7. Nursing Homes

    Science.gov (United States)

    ... Home › Aging & Health A to Z › Nursing Homes Font size A A A Print Share Glossary Basic ... Reason For Living in A Nursing Home Some type of disability with activities of daily living (ADLs) ...

  8. Home Care

    Science.gov (United States)

    ... are part of home healthcare agencies. You may benefit from home care if you are dealing with ... it will trigger an emergency response or checkup phone call. Newer technologies ... or mobile testing technology (home diagnostics), including x-rays and ...

  9. Relations among work-home culture, the utilization of work-home arrangements, and work-home interference

    NARCIS (Netherlands)

    Dikkers, J.S.E.; Geurts, S.A.E.; Dulk, L. den; Peper, A.B.; Kompier, M.A.J.

    2004-01-01

    In the present study, we examined the associations among work-home culture (WHC), the utilization of work-home arrangements (WHAs), and work-home interference (WHI) among 638 workers from a Dutch financial consultancy firm. We (a) developed a typology of WHC, (b) examined whether the utilization of

  10. Fluorescence quenching of graphene oxide combined with the site-specific cleavage of restriction endonuclease for deoxyribonucleic acid demethylase activity assay

    Energy Technology Data Exchange (ETDEWEB)

    Ji, Lijuan; Qian, Yingdan; Wu, Ping; Zhang, Hui; Cai, Chenxin, E-mail: cxcai@njnu.edu.cn

    2015-04-15

    Highlights: • An approach for sensitive and selective DNA demethylase activity assay is reported. • This assay is based on the fluorescence quenching of GO and site-specific cleavage of endonuclease. • It can determine as low as 0.05 ng mL{sup −1} of MBD2 with a linear range of 0.2–300 ng mL{sup −1}. • It has an ability to recognize MBD2 from other possibly coexisting proteins and cancer cell extracts. • It can avoid false signals, requiring no bisulfite conversion, PCR amplification, radioisotope-labeling. - Abstract: We report on the development of a sensitive and selective deoxyribonucleic acid (DNA) demethylase (using MBD2 as an example) activity assay by coupling the fluorescence quenching of graphene oxide (GO) with the site-specific cleavage of HpaII endonuclease to improve the selectivity. This approach was developed by designing a single-stranded probe (P1) that carries a binding region to facilitate the interaction with GO, which induces fluorescence quenching of the labeled fluorophore (FAM, 6-carboxyfluorescein), and a sensing region, which contains a hemi-methylated site of 5′-CmCGG-3′, to specifically recognize the target (T1, a 32-mer DNA from the promoter region of p53 gene) and hybridize with it to form a P1/T1 duplex. After demethylation with MBD2, the duplex can be specifically cleaved using HpaII, which releases the labeled FAM from the GO surface and results in the recovery of fluorescence. However, this cleavage is blocked by the hemi-methylation of this site. Thus, the magnitude of the recovered fluorescence signal is related to the MBD2 activity, which establishes the basis of the DNA demethylase activity assay. This assay can determine as low as ∼(0.05 ± 0.01) ng mL{sup −1} (at a signal/noise of 3) of MBD2 with a linear range of 0.2–300 ng mL{sup −1} and recognize MBD2 from other possibly coexisting proteins and cancer cell extracts. The advantage of this assay is its ability to avoid false signals and no

  11. Associations Between Home Death and the Use and Type of Care at Home.

    Science.gov (United States)

    McEwen, Rebecca; Asada, Yukiko; Burge, Frederick; Lawson, Beverley

    2018-01-01

    Despite wishes for and benefits of home deaths, a discrepancy between preferred and actual location of death persists. Provision of home care may be an effective policy response to support home deaths. Using the population-based mortality follow-back study conducted in Nova Scotia, we investigated the associations between home death and formal care at home and between home death and the type of formal care at home. We found (1) the use of formal care at home at the end of life was associated with home death and (2) the use of formal home support services at home was associated with home death among those whose symptoms were well managed.

  12. Home

    Science.gov (United States)

    Military Commission Seal VWAP Login Home Go ABOUT US Organization Overview Organizational Chart Families VWAP Login CCTV Sites Travel Media MC News CCTV Sites Travel Today at OMC Home Today at OMC Daily

  13. Home hemodialysis

    DEFF Research Database (Denmark)

    Agar, John W; Perkins, Anthony; Heaf, James G

    2015-01-01

    We describe the infrastructure that is necessary for hemodialysis in the home focusing on physical requirements, the organization of plumbing and water, and the key features that should guide the selection of machines that are suitable for home use.......We describe the infrastructure that is necessary for hemodialysis in the home focusing on physical requirements, the organization of plumbing and water, and the key features that should guide the selection of machines that are suitable for home use....

  14. HomePort

    DEFF Research Database (Denmark)

    Madsen, Per Printz

    2009-01-01

    In the last couple of year's computer based home control systems are getting more and more common in modern homes. For instance these systems take care of light control, heat control and security systems.  The latest trend is to use wireless communication like Z-Wave and ZigBee to interconnect...... different components in these systems. One of the characteristics is that each system, like for instance heat and light, has their own specific way of using the communication system.   This paper describes a way to connect different home control systems through an intelligent gateway, called a Home......Port. The HomePort consists of a number of Subsystem communication drivers, a virtual communication layer, an interpreter and a PC- based compiler for a high level control language, called GIL (Gateway intelligence language). The focus in this paper will be on the upper two layers in the Home...

  15. DOE Zero Energy Ready Home Case Study: Thrive Home Builders, Lowry Plan

    Energy Technology Data Exchange (ETDEWEB)

    Pacific Northwest National Laboratory

    2017-09-01

    Thrive Home Builders built this 4,119-ft2 home at the Lowry development in Denver, Colorado, to the high-performance criteria of the U.S. Department of Energy’s Zero Energy Ready Home Program. Despite the dense positioning of the homes, mono-plane roof designs afforded plenty of space for the 8.68 kW of photovoltaic panels. With the PV, the home achieves a Home Energy Rating System (HERS) score of 4 and the home owners should enjoy energy bills of about $-11 a year. Without the PV, the home would score a HERS 38 (far lower than the HERS 80 to 100 of typical new homes).

  16. Multidisciplinary nutritional support for undernutrition in nursing home and home-care

    DEFF Research Database (Denmark)

    Beck, Anne Marie; Gøgsig Christensen, Annette; Stenbæk Hansen, Birthe

    2016-01-01

    Objective To assess the effect of multidisciplinary nutritional support for undernutrition in older adults in nursing home and home-care identified with the validated Eating Validation Scheme (EVS). Methods An 11 wk cluster randomized trial with a home-care (3 clusters) or nursing home (3 clusters.......3] versus 1.3 [0.5], P = 0.021) was observed. There was a almost significant difference in mortality (2% versus 13%, P = 0.079). Conclusions Multidisciplinary nutritional support in older adults in nursing home and home-care could have a positive effect on quality of life, muscle strength, and oral care....... means of EuroQol-5D-3L), physical performance (30-seconds chair stand), nutritional status (weight and hand-grip strength), oral care, fall incidents, hospital admissions, rehabilitation stay, moving to nursing homes (participants from home-care), and mortality. Results Respectively, 55 (46 from 2 home...

  17. The Medical Home

    Science.gov (United States)

    ... Videos for Educators Search English Español The Medical Home KidsHealth / For Parents / The Medical Home What's in ... for your child. What Does the Term "Medical Home" Mean? A medical home isn't a place ...

  18. Home Rx: The Health Benefits of Home Performance

    Energy Technology Data Exchange (ETDEWEB)

    Wilson, Jonathan [National Center for Healthy Housing (NCHH), Columbia, MD (United States); Jacobs, David [National Center for Healthy Housing (NCHH), Columbia, MD (United States); Reddy, Amanda [National Center for Healthy Housing (NCHH), Columbia, MD (United States); Tohn, Ellen [Tohn Environmental Strategies, Wayland, MA (United States); Cohen, Jonathan [Dept. of Energy (DOE), Washington DC (United States); Jacobsohn, Ely [Dept. of Energy (DOE), Washington DC (United States)

    2016-12-01

    Evidence in a new, groundbreaking U.S. Department of Energy report, Home Rx: The Health Benefits of Home Performance, shows that home performance upgrades can improve the quality of a home’s indoor environment by reducing the prevalence of harmful indoor air pollutants and contaminants. Until recently, no systematic review of this evidence had been conducted, limiting full understanding of the link between home performance and health. This new report summarizes current knowledge and identifies research gaps. The design characteristics and results of each of the 40 studies considered in the report are summarized in a searchable matrix.

  19. ThinkHome Energy Efficiency in Future Smart Homes

    Directory of Open Access Journals (Sweden)

    Reinisch Christian

    2011-01-01

    Full Text Available Abstract Smart homes have been viewed with increasing interest by both home owners and the research community in the past few years. One reason for this development is that the use of modern automation technology in the home or building promises considerable savings of energy, therefore, simultaneously reducing the operational costs of the building over its whole lifecycle. However, the full potential of smart homes still lies fallow, due to the complexity and diversity of the systems, badly engineered and configured installations, as well as the frequent problem of suboptimal control strategies. Summarized, these problems converge to two undesirable conditions in the "not-so-smart" home: energy consumption is still higher than actually necessary and users are unable to yield full comfort in their automated homes. This work puts its focus on alleviating the current problems by proposing a comprehensive system concept, that shall ensure that smart homes can keep their promise in the future. The system operates on an extensive knowledge base that stores all information needed to fulfill the goals of energy efficiency and user comfort. Its intelligence is implemented as and within a multiagent system that also caters for the system's openness to the outside world. As a first evaluation, a profile-based control strategy for thermal comfort is developed and verified by means of simulation.

  20. Unimaginable homes

    DEFF Research Database (Denmark)

    Møller, Kristian; Klausen, Maja

    2018-01-01

    The chapter draw from critical mediatization theory, critical intimacy theory, and cultural gerontology and asks: How do elderly people practice their mediatized homes? Which roles do media play in constituting and disturbing the flows of bodies into the home? Moreover: how do dominant...... in the making of the mediatized home space. We conclude by returning to the research questions and making explicit how researching flows of bodies that in many ways inhabit homes of the in-between contributes to both gerontological and geomediatization research agendas....

  1. Stroke and Nursing Home care: a national survey of nursing homes.

    LENUS (Irish Health Repository)

    Cowman, Seamus

    2010-01-01

    BACKGROUND: Although stroke is recognised as a major factor in admission to nursing home care, data is lacking on the extent and nature of the disabilities and dependency in nursing homes arising from stroke. A national study conducted in nursing homes can quantify the number of residents with stroke in nursing homes, their disability and levels of dependency. METHODS: A cross-sectional survey research design was used. A total of 572 public and private nursing homes were identified nationally and a stratified random selection of 60 nursing homes with 3,239 residents was made. In half of the nursing homes (n = 30) efforts were made to interview all residents with stroke Survey instruments were used to collect data from residents with stroke and nursing home managers on demography, patient disability, and treatment. RESULTS: Across all nursing homes (n = 60), 18% (n = 570) of the residents had previously had a stroke. In homes (n = 30), where interviews with residents with stroke (n = 257), only 7% (n = 18) residents were capable of answering for themselves and were interviewed. Data on the remaining 93% (n = 239) residents were provided by the nursing home manager. Nurse Managers reported that 73% of residents with stroke had a high level of dependency. One in two residents with stroke was prescribed antidepressants or sedative medication. Only 21% of stroke residents were prescribed anticoagulants, 42% antiplatelets, and 36% cholesterol lowering medications. Stroke rehabilitation guidelines were lacking and 68% reported that there was no formal review process in place. CONCLUSIONS: This study provides seminal findings on stroke and nursing home services in Ireland. We now know that one in six nursing home residents in a national survey are residents with a stroke, and have a wide range of disabilities. There is currently little or no structured care (beyond generic care) for stroke survivors who reside in nursing homes in Ireland.

  2. Home Automation : Smart home technology and template house design

    OpenAIRE

    Zheng, Zeya

    2013-01-01

    In this thesis, home automation’s general knowledge, technology information and each component will be introduced to the reader in the first half of the whole thesis. In the second half, thesis includes the Home Automation template design and market competitiveness analysis. The author assumes that the reader is going to spend lots of money to have a smart home. In this situation, the author introduces the home automation to the reader at each component. So the reader in this thesis actu...

  3. Predictors of Home Care Expenditures and Death at Home for Cancer Patients in an Integrated Comprehensive Palliative Home Care Pilot Program

    Science.gov (United States)

    Howell, Doris M.; Abernathy, Tom; Cockerill, Rhonda; Brazil, Kevin; Wagner, Frank; Librach, Larry

    2011-01-01

    Purpose: Empirical understanding of predictors for home care service use and death at home is important for healthcare planning. Few studies have examined these predictors in the context of the publicly funded Canadian home care system. This study examined predictors for home care use and home death in the context of a “gold standard” comprehensive palliative home care program pilot in Ontario where patients had equal access to home care services. Methods: Secondary clinical and administrative data sources were linked using a unique identifier to examine multivariate factors (predisposing, enabling, need) on total home care expenditures and home death for a cohort of cancer patients enrolled in the HPCNet pilot. Results: Subjects with gastrointestinal symptoms (OR: 1.64; p=0.03) and those with higher income had increased odds of dying at home (OR: 1.14; phome care expenditures. Conclusions: Predictors of home death found in earlier studies appeared less important in this comprehensive palliative home care pilot. An income effect for home death observed in this study requires examination in future controlled studies. Relevance: Access to palliative home care that is adequately resourced and organized to address the multiple domains of issues that patients/families experience at the end of life has the potential to enable home death and shift care appropriately from limited acute care resources. PMID:22294993

  4. The GAN Exonuclease or the Flap Endonuclease Fen1 and RNase HII Are Necessary for Viability of Thermococcus kodakarensis.

    Science.gov (United States)

    Burkhart, Brett W; Cubonova, Lubomira; Heider, Margaret R; Kelman, Zvi; Reeve, John N; Santangelo, Thomas J

    2017-07-01

    Many aspects of and factors required for DNA replication are conserved across all three domains of life, but there are some significant differences surrounding lagging-strand synthesis. In Archaea , a 5'-to-3' exonuclease, related to both bacterial RecJ and eukaryotic Cdc45, that associates with the replisome specifically through interactions with GINS was identified and designated GAN (for G INS- a ssociated n uclease). Despite the presence of a well-characterized flap endonuclease (Fen1), it was hypothesized that GAN might participate in primer removal during Okazaki fragment maturation, and as a Cdc45 homologue, GAN might also be a structural component of an archaeal CMG (Cdc45, MCM, and GINS) replication complex. We demonstrate here that, individually, either Fen1 or GAN can be deleted, with no discernible effects on viability and growth. However, deletion of both Fen1 and GAN was not possible, consistent with both enzymes catalyzing the same step in primer removal from Okazaki fragments in vivo RNase HII has also been proposed to participate in primer processing during Okazaki fragment maturation. Strains with both Fen1 and RNase HII deleted grew well. GAN activity is therefore sufficient for viability in the absence of both RNase HII and Fen1, but it was not possible to construct a strain with both RNase HII and GAN deleted. Fen1 alone is therefore insufficient for viability in the absence of both RNase HII and GAN. The ability to delete GAN demonstrates that GAN is not required for the activation or stability of the archaeal MCM replicative helicase. IMPORTANCE The mechanisms used to remove primer sequences from Okazaki fragments during lagging-strand DNA replication differ in the biological domains. Bacteria use the exonuclease activity of DNA polymerase I, whereas eukaryotes and archaea encode a flap endonuclease (Fen1) that cleaves displaced primer sequences. RNase HII and the GINS-associated exonuclease GAN have also been hypothesized to assist in primer

  5. Eldercare at Home: Choosing a Nursing Home

    Science.gov (United States)

    ... required, these services can be provided by a separate home health agency as directed by a doctor or ... complaints made by or on behalf of nursing home residents and work to resolve the problems. If they are unable ...

  6. Smart Home Hacking

    OpenAIRE

    Kodra, Suela

    2016-01-01

    Smart Home is an intelligent home equipped with devices and communications systems that enables the residents to connect and control their home appliances and systems. This technology has changed the way a consumer interacts with his home, enabling more control and convenience. Another advantage of this technology is the positive impact it has on savings on energy and other resources. However, despite the consumer's excitement about smart home, security and privacy have shown to be the strong...

  7. Study on detection of mutation DNA fragment in gastric cancer by restriction endonuclease fingerprinting with capillary electrophoresis.

    Science.gov (United States)

    Wang, Rong; Xie, Hua; Xu, Yue-Bing; Jia, Zheng-Ping; Meng, Xian-Dong; Zhang, Juan-Hong; Ma, Jun; Wang, Juan; Wang, Xian-Hua

    2012-03-01

    The DNA fragment detection focusing technique has further enhanced the sensitivity and information of DNA targets. The DNA fragment detection method was established by capillary electrophoresis with laser-induced fluorescence detection and restriction endonuclease chromatographic fingerprinting (CE-LIF-REF) in our experiment. The silica capillary column was coated with short linear polyarclarylamide (SLPA) using nongel sieving technology. The excision product of various restricted enzymes of DNA fragments was obtained by REF with the molecular biology software Primer Premier 5. The PBR322/BsuRI DNA marker was used to establish the optimization method. The markers were focused electrophoretically and detected by CE-LIF. The results demonstrate that the CE-LIF-REF with SLPA can improve separation, sensitivity and speed of analysis. This technique may be applied to analysis of the excision product of various restricted enzymes of prokaryotic plasmid (pIRES2), eukaryote plasmid (pcDNA3.1) and the PCR product of codon 248 region of gastric cancer tissue. The results suggest that this method could very sensitively separate the excision products of various restricted enzymes at a much better resolution than the traditional agarose electrophoresis. Copyright © 2011 John Wiley & Sons, Ltd.

  8. Stroke and Nursing Home care: a national survey of nursing homes

    Directory of Open Access Journals (Sweden)

    McGee Hannah

    2010-01-01

    Full Text Available Abstract Background Although stroke is recognised as a major factor in admission to nursing home care, data is lacking on the extent and nature of the disabilities and dependency in nursing homes arising from stroke. A national study conducted in nursing homes can quantify the number of residents with stroke in nursing homes, their disability and levels of dependency. Methods A cross-sectional survey research design was used. A total of 572 public and private nursing homes were identified nationally and a stratified random selection of 60 nursing homes with 3,239 residents was made. In half of the nursing homes (n = 30 efforts were made to interview all residents with stroke Survey instruments were used to collect data from residents with stroke and nursing home managers on demography, patient disability, and treatment. Results Across all nursing homes (n = 60, 18% (n = 570 of the residents had previously had a stroke. In homes (n = 30, where interviews with residents with stroke (n = 257, only 7% (n = 18 residents were capable of answering for themselves and were interviewed. Data on the remaining 93% (n = 239 residents were provided by the nursing home manager. Nurse Managers reported that 73% of residents with stroke had a high level of dependency. One in two residents with stroke was prescribed antidepressants or sedative medication. Only 21% of stroke residents were prescribed anticoagulants, 42% antiplatelets, and 36% cholesterol lowering medications. Stroke rehabilitation guidelines were lacking and 68% reported that there was no formal review process in place. Conclusions This study provides seminal findings on stroke and nursing home services in Ireland. We now know that one in six nursing home residents in a national survey are residents with a stroke, and have a wide range of disabilities. There is currently little or no structured care (beyond generic care for stroke survivors who reside in nursing homes in Ireland.

  9. Species attribution and strain typing of Oenococcus oeni (formerly Leuconostoc oenos) with restriction endonuclease fingerprints.

    Science.gov (United States)

    Viti, C; Giovannetti, L; Granchi, L; Ventura, S

    1996-10-01

    In several wines, malolactic fermentation is required to improve the organoleptic characters and to stabilize the final product. In order to establish a controlled malolactic fermentation in wine, easy identification and sensitive typing of strains of Oenococcus oeni (new name of the malolactic bacterium Leuconostoc oenos) used as starter cultures are necessary. To accomplish these tasks, several strains of Oenococcus oeni isolated from wines of the Chianti region (Italy), along with reference strains and strains of L. mesenteroides subsp. mesenteroides, L. carnosum, L. fallax, L. pseudomesenteroides, L. lactis and Weisella paramesenteroides, were studied with RFLP of ribosomal genes and ultrasensitive total DNA restriction pattern analysis performed on polyacrylamide gel. With each of four restriction endonucleases used, identical restriction profiles of ribosomal genes were obtained for all strains of O. oeni. These ribopatterns, being strongly dissimilar to profiles of the other lactic acid bacteria tested, appear to be well suited for the attribution of wine lactic acid bacteria to the species O. oeni. Cluster analysis performed on two total DNA restriction profile data sets showed that the species O. oeni possesses a good degree of genomic homogeneity. Very sensitive typing of strains of O. oeni was obtained with total DNA restriction profiles. The potential of an integrated approach using restriction profiles for species assignment and typing of selected malolactic bacteria is demonstrated.

  10. Mitochondrial Targeted Endonuclease III DNA Repair Enzyme Protects against Ventilator Induced Lung Injury in Mice

    Directory of Open Access Journals (Sweden)

    Masahiro Hashizume

    2014-08-01

    Full Text Available The mitochondrial targeted DNA repair enzyme, 8-oxoguanine DNA glycosylase 1, was previously reported to protect against mitochondrial DNA (mtDNA damage and ventilator induced lung injury (VILI. In the present study we determined whether mitochondrial targeted endonuclease III (EndoIII which cleaves oxidized pyrimidines rather than purines from damaged DNA would also protect the lung. Minimal injury from 1 h ventilation at 40 cmH2O peak inflation pressure (PIP was reversed by EndoIII pretreatment. Moderate lung injury due to ventilation for 2 h at 40 cmH2O PIP produced a 25-fold increase in total extravascular albumin space, a 60% increase in W/D weight ratio, and marked increases in MIP-2 and IL-6. Oxidative mtDNA damage and decreases in the total tissue glutathione (GSH and the GSH/GSSH ratio also occurred. All of these indices of injury were attenuated by mitochondrial targeted EndoIII. Massive lung injury caused by 2 h ventilation at 50 cmH2O PIP was not attenuated by EndoIII pretreatment, but all untreated mice died prior to completing the two hour ventilation protocol, whereas all EndoIII-treated mice lived for the duration of ventilation. Thus, mitochondrial targeted DNA repair enzymes were protective against mild and moderate lung damage and they enhanced survival in the most severely injured group.

  11. Medicare home health utilization as a function of nursing home market factors.

    OpenAIRE

    Swan, J H; Benjamin, A E

    1990-01-01

    Rapid increases in the size and costs of the home health market, unknown impacts of Medicare's DRG hospital reimbursement on the posthospital market, and general lack of knowledge about factors that explain interstate variation in home health utilization all suggest the importance of developing and testing models of Medicare home health use. This article proposes and tests a model of state home health utilization as a function of the nursing home market. This model proposes that home health u...

  12. The Sweet-Home speech and multimodal corpus for home automation interaction

    OpenAIRE

    Vacher , Michel; Lecouteux , Benjamin; Chahuara , Pedro; Portet , François; Meillon , Brigitte; Bonnefond , Nicolas

    2014-01-01

    International audience; Ambient Assisted Living aims at enhancing the quality of life of older and disabled people at home thanks to Smart Homes and Home Automation. However, many studies do not include tests in real settings, because data collection in this domain is very expensive and challenging and because of the few available data sets. The SWEET-H OME multimodal corpus is a dataset recorded in realistic conditions in D OMUS, a fully equipped Smart Home with microphones and home automati...

  13. Smoke-Free Homes and Home Exposure to Secondhand Smoke in Shanghai, China

    Directory of Open Access Journals (Sweden)

    Pinpin Zheng

    2014-11-01

    Full Text Available Few studies have examined home exposure to secondhand smoke (SHS in China. This study aimed to document: (1 the prevalence and correlates of exposure to SHS in homes (in adult non-smokers in Shanghai, and (2 enforcement of rules, harm reduction behaviors, and self-efficacy for maintaining smoke-free homes in Shanghai. A total of 500 participants were recruited using a multistage proportional random sampling design in an urban and suburban district to complete a survey. Among the total 355 nonsmokers, 127 (35.8% participants reported being exposed to SHS in the past 7 days. Participants living with smokers in the home, with no smoking restriction at home, and having children younger than 18 were more likely to be exposed to SHS at home. Higher self-efficacy in maintaining a smoke-free home was negatively associated with home SHS exposure. Having visitors who smoke was the greatest policy enforcement challenge. Ineffective measures such as opening windows were more commonly used in homes with partial bans. Educational initiatives to protect against SHS exposure in the home should promote smoke-free homes, address challenges to implementing such policies, and address misconceptions regarding the effectiveness of supposed harm reduction behaviors.

  14. DOE Zero Energy Ready Home Case Study: Sterling Brook Custom Homes — Village Park Eco Home, Double Park, TX

    Energy Technology Data Exchange (ETDEWEB)

    none,

    2014-09-01

    This builder won a Custom Builder honor in the 2014 Housing Innovation Awards for this showcase home that serves as an energy-efficient model home for the custom home builder: 1,300 visitors toured the home, thousands more learned about the home’s advanced construction via the webpage, YouTube, Twitter, Facebook, Instagram, and Pinterest.

  15. HOME Grantee Areas

    Data.gov (United States)

    Department of Housing and Urban Development — The HOME Investment Partnership Program (HOME) is authorized under Title II of the Cranston-Gonzalez National Affordable Housing Act. HOME provides formula grants to...

  16. Bonding mechanism of a yttrium iron garnet film on Si without the use of an intermediate layer

    International Nuclear Information System (INIS)

    Pantzas, Konstantinos; Patriarche, Gilles; Talneau, Anne; Youssef, Jamal Ben

    2014-01-01

    Direct bonding of yttrium iron garnet (YIG) on silicon without the use of an intermediate bonding layer is demonstrated and characterized using scanning transmission electron microscopy and energy-dispersive x-ray spectroscopy. During the bonding experiment, the garnet is reduced in the presence of oxide-free silicon. As a result, a 5 nm thick SiO 2 /amorphous-YIG bilayer is formed and welds the garnet to silicon.

  17. Smart Cities - Smart Homes and Smart Home Technology

    OpenAIRE

    Faanes, Erlend Kydland

    2014-01-01

    This master’s thesis consists of two articles where the first article is theoretical and the second is the empirical study. Article I The purpose with this paper is to explore and illuminate how smart home and smart home technology can contribute to enhance health and Quality of Life in elderly citizens and allow them to live longer in their home. The paper provides a brief introduction to health promotion and highlights the thesis theoretical framework and foundation of Aaron Antonov...

  18. Nursing perception of patient transitions from hospitals to home with home health.

    Science.gov (United States)

    Smith, Shannon Bright; Alexander, Judith W

    2012-01-01

    The study's purpose was to determine nurses' opinions of sending patients from the hospital to home with home health services. The study occurred in the Charleston, South Carolina, Tricounty area (Berkeley, Charleston, and Dorchester counties). Home health agencies and hospitals were invited to participate. The study used a survey design to gather information on nursing perceptions of current practices and needed changes to improve transition of patients. The population was nurses (licensed practical nurses (LPNs) and registered nurses (RNs)) employed at inpatient hospitals or home health agencies in the area. Thirty-four RNs responded with no LPNs respondents. Agency administrators/chief nursing officers agreed for their agencies to participate and distributed the survey using a Research Electronic Data Capture (REDCap) Internet-based survey tool. Using the survey results and information from a literature review, the study developed a list of propositions, which participating administrators reviewed, for improving transitions to home. Both home health and hospital nurses reported a need to improve the process of sending patients from hospital to home with home health services. This study provides hospitals and home health agencies with propositions to facilitate the establishment of a process to communicate effectively patients care needs and streamline the discharging patients from the hospital to home health care; thus, improving patient transition. Case managers and discharge planners will need interagency collaboration along with evidence-based interventions to transition patients from the hospital to home with home health services with various populations. Direct patient care nurses in both hospital and home health settings should share the same accountability as case managers to ensure successful transitions.

  19. Representing in-home and out-of-home energy consumption behavior in Beijing

    International Nuclear Information System (INIS)

    Yu Biying; Zhang Junyi; Fujiwara, Akimasa

    2011-01-01

    It is expected that in-home and out-of-home energy consumption behavior in a household might be correlated with each other, probably due to the existence of household budget constraints. Ownership and usage of energy-saving technologies for in-home appliances (or vehicles) might lead to the increase in out-of-home (or in-home) energy consumption. It is therefore necessary to jointly represent in-home and out-of-home energy consumption in the same modeling framework. With this consideration, we first build a new type of energy consumption model based on the Multiple Discrete-Continuous Extreme Value (MDCEV) modeling framework. Next, we conducted a questionnaire survey in Beijing in 2009 and successfully collected the information about households' energy consumption, ownership/usage of in-home appliances and vehicles, and households' and their members' attributes from 1014 households. Throughout an empirical analysis, it is confirmed that the MDCEV model is effective to simultaneously describe the in-home and out-of-home energy consumption behavior. In addition, it is revealed that a set of household and personal attributes affect the ownership and usage of in-home appliances and vehicles. Furthermore, it is shown that the unobserved factors play a much more important role in explaining energy consumption behavior than the observed attributes of households and their members. - Highlights: → Representing in-home and out-of-home energy consumption behavior jointly. → MDCEV model is built to describe household energy consumption behavior. → Log-linear competitive relationships are found among expenditures of end-uses. → Model results provide some insights about the influence of varied observed factors. → Unobserved factors are more important in explaining energy consumption behavior.

  20. Home Health Compare: Find a Home Health Agency

    Science.gov (United States)

    ... page could not be loaded. The Medicare.gov Home page currently does not fully support browsers with " ... widget - Select to show Back to top Footer Home A federal government website managed and paid for ...

  1. Exercise at Home

    Science.gov (United States)

    ... Home Health Insights Exercise & Weight Exercise at Home Exercise at Home Make an Appointment Ask a Question ... with the movement and contact your provider. Posture Exercises Better posture means better breathing and movement. Axial ...

  2. Home, Smart Home

    DEFF Research Database (Denmark)

    Hansen, Ellen Kathrine; Olesen, Gitte Gylling Hammershøj; Mullins, Michael

    2013-01-01

    The article places focus on how smart technologies integrated in a one family- home and particular the window offer unique challenges and opportunities for designing buildings with the best possible environments for people and nature. Toward an interdisciplinary approach, we address the interaction...... between daylight defined in technical terms and daylight defined in aesthetic, architectural terms. Through field-tests of a Danish carbon-neutral home and an analysis of five key design parameters, we explore the contradictions and potentials in smart buildings, using the smart window as example of how...... to the energy design is central. The study illuminates an approach of the design of smart houses as living organisms by connecting technology with the needs of the occupants with the power and beauty of daylight....

  3. Bringing Work Home: Take-Home Pesticide Exposure Among Farm Families

    NARCIS (Netherlands)

    Curwin, B.D.

    2006-01-01

    In this thesis take-home pesticide exposure among farm families, with an emphasis on herbicides, was investigated. Take-home exposure occurs when a worker unwittingly brings home a substance on his or her clothing or shoes, thereby potentially exposing his or her family. The pesticides investigated

  4. In-home contextual reality: a qualitative analysis using the Multiple Errands Test Home Version (MET-Home).

    Science.gov (United States)

    Burns, Suzanne Perea; Pickens, Noralyn Davel; Dawson, Deirdre R; Perea, Jaimee D; Vas, Asha K; Marquez de la Plata, Carlos; Neville, Marsha

    2018-02-06

    Adults with stroke frequently experience executive dysfunction. Despite the range of assessments that examine the effects of executive dysfunction on daily tasks, there remains a paucity of literature that examines the influence of the environment on performance in the community. The MET-Home is an ecologically valid assessment for examining post-stroke executive dysfunction in the home environment. This qualitative study explores the relationship between the environment and MET-Home performance among home-dwelling adults with stroke and matched controls. Using a descriptive qualitative approach, we analysed video, interview, and observation notes from a MET-Home validation study. An overarching theme of interplay between everyday task performance and the home environment produced further themes: naturalistically emerging supports and barriers and environment as strategy. Within naturalistically emerging supports and barriers, five contextual sub-themes were discovered: physical environment, social environment, temporal context, virtual context, and personal context. Within environment as strategy, we identified four sub-themes: reducing distractions, using everyday technologies, planning in context, and seeking social support. These findings extend the conceptualisation of how we evaluate executive dysfunction in the context of the community to also consider the inherent influence of the environment.

  5. Haplotype-based case-control study between human apurinic/apyrimidinic endonuclease 1/redox effector factor-1 gene and cerebral infarction.

    Science.gov (United States)

    Naganuma, Takahiro; Nakayama, Tomohiro; Sato, Naoyuki; Fu, Zhenyan; Yamaguchi, Mai; Soma, Masayoshi; Aoi, Noriko; Usami, Ron; Doba, Nobutaka; Hinohara, Shigeaki

    2009-10-01

    The aim of this study was to investigate the relationship between cerebral infarction (CI) and the human apurinic/apyrimidinic endonuclease 1/redox effector factor-1 (APE1/REF-1) gene using single-nucleotide polymorphisms (SNPs) and a haplotype-based case-control study. We selected 5 SNPs in the human APE1/REF1 gene (rs1760944, rs3136814, rs17111967, rs3136817 and rs1130409), and performed case-control studies in 177 CI patients and 309 control subjects. rs17111967 was found to have no heterogeneity in Japanese. The overall distribution of the haplotype-based case-control study constructed by rs1760944, rs3136814 and rs1130409 showed a significant difference. The frequency of the G-C-T haplotype was significantly higher in the CI group than in the control group (2.5% vs. 0.0%, p>0.001). Based on the results of the haplotype-based case-control-study, the G-C-T haplotype may be a genetic marker of CI, and the APE1/REF-1 gene may be a CI susceptibility gene.

  6. Bulk magnon spin current theory for the longitudinal spin Seebeck effect

    Energy Technology Data Exchange (ETDEWEB)

    Rezende, S.M., E-mail: rezende@df.ufpe.br [Departamento de Física, Universidade Federal de Pernambuco, 50670-901 Recife, Pernambuco (Brazil); Rodríguez-Suárez, R.L. [Departamento de Física, Universidade Federal de Pernambuco, 50670-901 Recife, Pernambuco (Brazil); Facultad de Física, Pontificia Universidad Católica de Chile, Casilla, 306 Santiago (Chile); Cunha, R.O.; López Ortiz, J.C.; Azevedo, A. [Departamento de Física, Universidade Federal de Pernambuco, 50670-901 Recife, Pernambuco (Brazil)

    2016-02-15

    The longitudinal spin Seebeck effect (LSSE) consists in the generation of a spin current parallel to a temperature gradient applied across the thickness of a bilayer made of a ferromagnetic insulator (FMI), such as yttrium iron garnet (YIG), and a metallic layer (ML) with strong spin orbit coupling, such as platinum. The LSSE is usually detected by a DC voltage generated along the ML due to the conversion of the spin current into a charge current perpendicular to the static magnetic field by means of the inverse spin Hall effect. Here we present a model for the LSSE that relies on the bulk magnon spin current created by the temperature gradient across the thickness of the FMI. We show that the spin current pumped into the metallic layer by the magnon accumulation in the FMI provides continuity of the spin current at the FMI/ML interface and is essential for the existence of the LSSE. The results of the theory are in good agreement with experimental LSSE data in YIG/Pt bilayers on the variation of the DC voltage with the sample temperature, with the FMI layer thickness and with the intensity of high magnetic fields. - Highlights: • We present a theory for the longitudinal spin Seebeck effect based on bulk magnons. • The model explains quantitatively the measured voltage in YIG/Pt created by the LSSE. • The model explains quantitatively the temperature dependence of LSSE measured in YIG/Pt. • The model agrees qualitatively with the measured dependence of LSSE with YIG thickness. • The model agrees qualitatively with the measured dependence of LSSE on magnetic field.

  7. Smart homes and home health monitoring technologies for older adults: A systematic review.

    Science.gov (United States)

    Liu, Lili; Stroulia, Eleni; Nikolaidis, Ioanis; Miguel-Cruz, Antonio; Rios Rincon, Adriana

    2016-07-01

    Around the world, populations are aging and there is a growing concern about ways that older adults can maintain their health and well-being while living in their homes. The aim of this paper was to conduct a systematic literature review to determine: (1) the levels of technology readiness among older adults and, (2) evidence for smart homes and home-based health-monitoring technologies that support aging in place for older adults who have complex needs. We identified and analyzed 48 of 1863 relevant papers. Our analyses found that: (1) technology-readiness level for smart homes and home health monitoring technologies is low; (2) the highest level of evidence is 1b (i.e., one randomized controlled trial with a PEDro score ≥6); smart homes and home health monitoring technologies are used to monitor activities of daily living, cognitive decline and mental health, and heart conditions in older adults with complex needs; (3) there is no evidence that smart homes and home health monitoring technologies help address disability prediction and health-related quality of life, or fall prevention; and (4) there is conflicting evidence that smart homes and home health monitoring technologies help address chronic obstructive pulmonary disease. The level of technology readiness for smart homes and home health monitoring technologies is still low. The highest level of evidence found was in a study that supported home health technologies for use in monitoring activities of daily living, cognitive decline, mental health, and heart conditions in older adults with complex needs. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  8. Novel magnetic–dielectric composite ceramic obtained from Y{sub 3}Fe{sub 5}O{sub 12} and CaTiO{sub 3}

    Energy Technology Data Exchange (ETDEWEB)

    Paiva, D.V.M. [Grupo de Química de Materiais Avançados (GQMAT), Departamento de Química Analítica e Físico-Química, Universidade Federal do Ceará – UFC, Campus do Pici, CP 12100, CEP 60451-970 Fortaleza, CE (Brazil); Silva, M.A.S. [Laboratório de Telecomunicações e Ciência e Engenharia de Materiais (LOCEM), Departamento de Física, Universidade Federal do Ceará (Brazil); Ribeiro, T.S.; Vasconcelos, I.F. [Laboratório de Magnetismo e Materiais Magnéticos, Departamento de Engenharia Metalúrgica e de Materiais, UFC (Brazil); Sombra, A.S.B.; Góes, J.C. [Laboratório de Telecomunicações e Ciência e Engenharia de Materiais (LOCEM), Departamento de Física, Universidade Federal do Ceará (Brazil); Fechine, P.B.A., E-mail: fechine@ufc.br [Grupo de Química de Materiais Avançados (GQMAT), Departamento de Química Analítica e Físico-Química, Universidade Federal do Ceará – UFC, Campus do Pici, CP 12100, CEP 60451-970 Fortaleza, CE (Brazil)

    2015-09-25

    Highlights: • The density of composites was improved with addition of CTO. • Dielectric permittivity of the composites presented values above the expected. • The composites presented magnetic permeability higher than YIG after 500 MHz. - Abstract: This work investigates the microstructure and magnetic–dielectric properties of the CaTiO{sub 3} (CTO) and Y{sub 3}Fe{sub 5}O{sub 12} (YIG) composite ceramic. CTO is a paramagnetic ceramic, presents a positive value of Resonant Frequency Temperature Coefficients (τ{sub f}) and has high dielectric permittivity (ε{sub r}) while YIG is ferromagnetic, has low ε{sub r} and negative τ{sub f} values. Therefore, it is expected that the composite from these individual phase shows new properties, or a mix of them derived from each component. X-ray powder diffraction (XRPD), Scanning Electron Microscope (SEM), Raman and {sup 57}Fe Mössbauer spectroscopy were performed to confirm the phases of the composites. The dielectric and magnetic properties in Radio-Microwave frequencies were studied by impedance spectroscopy using different geometries. It was observed a densification improvement resulting from the insertion of the CTO in composites and a chemical reaction between YIG and CTO phases. This new composite has potential applications in bulk and thick/thin films devices.

  9. Meals in nursing homes

    DEFF Research Database (Denmark)

    Kofod, Jens Erik; Birkemose, A.

    2004-01-01

    Undernutrition is present among 33% of nursing home residents in Denmark. Hence, it is relevant to examine the meal situation at nursing homes to single out factors that may increase or reduce the residents' food intake. in the ongoing Danish nursing home debate it is claimed that a new type...... of nursing home improves the residents' meal situation with a positive effect on nutrition. The aim of this work is to test the general hypothesis that (i) residents appreciate the meal situation in these nursing homes and (ii) nutritional status of the residents is improved in this type of nursing home....... This study was carried out in four Danish nursing homes at various locations in Denmark. The methods used are qualitative interviews and observations at four nursing homes in combination with measurement of body mass index (BMI) at two of the four nursing homes. Undernutrition is defined as a BMI below 20...

  10. Home Care Services

    Science.gov (United States)

    Home care is care that allows a person with special needs stay in their home. It might be for people who are getting ... are chronically ill, recovering from surgery, or disabled. Home care services include Personal care, such as help ...

  11. Spin wave propagation in perpendicularly magnetized nm-thick yttrium iron garnet films

    Science.gov (United States)

    Chen, Jilei; Heimbach, Florian; Liu, Tao; Yu, Haiming; Liu, Chuanpu; Chang, Houchen; Stückler, Tobias; Hu, Junfeng; Zeng, Lang; Zhang, Youguang; Liao, Zhimin; Yu, Dapeng; Zhao, Weisheng; Wu, Mingzhong

    2018-03-01

    Magnonics offers a new way for information transport that uses spin waves (SWs) and is free of charge currents. Unlike Damon-Eshbach SWs, the magneto-static forward volume SWs offer the reciprocity configuration suitable for SW logic devices with low power consumption. Here, we study forward volume SW propagation in yttrium iron garnet (YIG) thin films with an ultra-low damping constant α = 8 ×10-5 . We design different integrated microwave antenna with different k-vector excitation distributions on YIG thin films. Using a vector network analyzer, we measured SW transmission with the films magnetized in perpendicular orientation. Based on the experimental results, we extract the group velocity as well as the dispersion relation of SWs and directly compare the power efficiency of SW propagation in YIG using coplanar waveguide and micro stripline for SW excitation and detection.

  12. Attitudes of palliative home care physicians towards palliative sedation at home in Italy.

    Science.gov (United States)

    Mercadante, Sebastiano; Masedu, Francesco; Mercadante, Alessandro; Marinangeli, Franco; Aielli, Federica

    2017-05-01

    Information about the attitudes towards palliative sedation (PS) at home is limited. The aim of this survey was to assess the attitudes of palliative care physicians in Italy regarding PS at home. A questionnaire was submitted to a sample of palliative care physicians, asking information about their activity and attitudes towards PS at home. This is a survey of home care physicians in Italy who were involved in end-of-life care decisions at home. One hundred and fifty participants responded. A large heterogeneity of home care organizations that generate some problems was found. Indications, intention and monitoring of PS seem to be appropriate, although some cultural and logistic conditions were limiting the use of PS. Specialized home care physicians are almost involved to start PS at home. Midazolam was seldom available at home and opioids were more frequently used. These data should prompt health care agencies to make a minimal set of drugs easily available for home care. Further research is necessary to compare attitudes in countries with different sociocultural profiles.

  13. Bringing Your Baby Home

    Science.gov (United States)

    ... for Educators Search English Español Bringing Your Baby Home KidsHealth / For Parents / Bringing Your Baby Home What's ... recall your baby's seemingly endless crying episodes. The Home Front Introducing your baby to others at home ...

  14. 24 CFR 982.610 - Group home: Who may reside in a group home.

    Science.gov (United States)

    2010-04-01

    ... 24 Housing and Urban Development 4 2010-04-01 2010-04-01 false Group home: Who may reside in a group home. 982.610 Section 982.610 Housing and Urban Development Regulations Relating to Housing and... Housing Types Group Home § 982.610 Group home: Who may reside in a group home. (a) An elderly person or a...

  15. Home area networks

    NARCIS (Netherlands)

    Koonen, A.M.J.

    2013-01-01

    This article consists of a collection of slides from the author's conference presentation. Some of the specific areas/topics discussed include: Convergence in home networks, home service scenarios; Home wired network architectures, CapEx and OpEx; Residential Gateway; Optical fiber types;

  16. Isolation and properties of the acid site-specific endonuclease from mature eggs of the sea urchin Strongylocentrotus intermedius

    International Nuclear Information System (INIS)

    Sibirtsev, Yu.T.; Konechnyi, A.A.; Rasskazov, V.A.

    1986-01-01

    An acid site-specific endonuclease has been detected in mature sea urchin eggs and cells of embryos at early stages of differentiation. Fractionation with ammonium sulfate, followed by chromatography on columns with DEAE, phosphocellulose, and hydroxyapatite resulted in an 18,000-fold purification. The molecular weight of the enzyme was determined at ∼ 29,000, the optimum pH 5.5. The activity of the enzyme does not depend on divalent metal ions, EDTA, ATP, and tRNA, but it is modulated to a substantial degree by NaCl. The maximum rate of cleavage of the DNA supercoil (form I) is observed at 100 mM NaCl. Increasing the NaCl concentration to 350 mM only slightly lowers the rate of cleavage of form I, yielding form II, but entirely suppresses the accumulation of form III. Restriction analysis of the products of enzymatic hydrolysis of Co1E1 and pBR322 DNA showed that at the early stages of hydrolysis the enzyme exhibits pronounced specificity for definite sites, the number of which is 12 for Co1 E1 DNA and 8 sites for pBR322 DNA

  17. Double-stranded endonuclease activity in Bacillus halodurans clustered regularly interspaced short palindromic repeats (CRISPR)-associated Cas2 protein.

    Science.gov (United States)

    Nam, Ki Hyun; Ding, Fran; Haitjema, Charles; Huang, Qingqiu; DeLisa, Matthew P; Ke, Ailong

    2012-10-19

    The CRISPR (clustered regularly interspaced short palindromic repeats) system is a prokaryotic RNA-based adaptive immune system against extrachromosomal genetic elements. Cas2 is a universally conserved core CRISPR-associated protein required for the acquisition of new spacers for CRISPR adaptation. It was previously characterized as an endoribonuclease with preference for single-stranded (ss)RNA. Here, we show using crystallography, mutagenesis, and isothermal titration calorimetry that the Bacillus halodurans Cas2 (Bha_Cas2) from the subtype I-C/Dvulg CRISPR instead possesses metal-dependent endonuclease activity against double-stranded (ds)DNA. This activity is consistent with its putative function in producing new spacers for insertion into the 5'-end of the CRISPR locus. Mutagenesis and isothermal titration calorimetry studies revealed that a single divalent metal ion (Mg(2+) or Mn(2+)), coordinated by a symmetric Asp pair in the Bha_Cas2 dimer, is involved in the catalysis. We envision that a pH-dependent conformational change switches Cas2 into a metal-binding competent conformation for catalysis. We further propose that the distinct substrate preferences among Cas2 proteins may be determined by the sequence and structure in the β1-α1 loop.

  18. Facilitating home birth.

    Science.gov (United States)

    Finigan, Valerie; Chadderton, Diane

    2015-06-01

    The birth of a baby is a family experience. However, in the United Kingdom birth often occurs outside the family environment, in hospital. Both home and hospital births have risks and benefits, but research shows that, for most women, it is as safe to give birth at home as it is in hospital. Women report home-birth to be satisfying with lowered risks of intervention and less likelihood of being separated from their family. It is also more cost effective for the National Health Service. Yet, whilst midwives are working hard to promote home birth as an option, it remains controversial. The aim of this paper is to raise awareness of the safety of home birth and the needs of women and midwives when a home birth is chosen. It provides an overview of care required and the role of the midwife in the ensuring care is woman-centred and personalised.

  19. Ultrasensitive electrochemical biosensor for detection of DNA from Bacillus subtilis by coupling target-induced strand displacement and nicking endonuclease signal amplification.

    Science.gov (United States)

    Hu, Yuhua; Xu, Xueqin; Liu, Qionghua; Wang, Ling; Lin, Zhenyu; Chen, Guonan

    2014-09-02

    A simple, ultrasensitive, and specific electrochemical biosensor was designed to determine the given DNA sequence of Bacillus subtilis by coupling target-induced strand displacement and nicking endonuclease signal amplification. The target DNA (TD, the DNA sequence from the hypervarient region of 16S rDNA of Bacillus subtilis) could be detected by the differential pulse voltammetry (DPV) in a range from 0.1 fM to 20 fM with the detection limit down to 0.08 fM at the 3s(blank) level. This electrochemical biosensor exhibits high distinction ability to single-base mismatch, double-bases mismatch, and noncomplementary DNA sequence, which may be expected to detect single-base mismatch and single nucleotide polymorphisms (SNPs). Moreover, the applicability of the designed biosensor for detecting the given DNA sequence from Bacillus subtilis was investigated. The result obtained by electrochemical method is approximately consistent with that by a real-time quantitative polymerase chain reaction detecting system (QPCR) with SYBR Green.

  20. Role of alkaline endonucleases in the release of soluble chromatin from thymus, spleen and liver nuclei of normal and irradiated mice

    International Nuclear Information System (INIS)

    Suciu, D.

    1979-01-01

    Thymus, spleen and liver nuclei released a large fraction of soluble chromatin in vitro when incubation was carried out in sucrose media containing low concentrations of CaCl 2 and/or MgCl 2 . A significant fraction of deoxyribopolynucleotides (DPN) was also extracted from nuclei. After 30 min of incubation at 37 0 C, the maximum release of soluble chromatin was observed near a pH of 8, which corresponds to the optimum pH of the alkaline endonuclease activity from thymus, spleen and liver. The soluble chromatin and DPN were precipitated by increasing the bivalent ion concentration of the medium. The protein/DNA ratio and the molecular weight of DNA suggest that the soluble chromatin and DPN represent nucleosome-like particles. The release of soluble chromatin in the first 4 hours of incubation was significantly increased if the nuclear fraction was isolated from the thymus and spleen of whole-body irradiated mice (1000 rad). This effect was absent in the liver nuclei. (author)

  1. Home range and travels

    Science.gov (United States)

    Stickel, L.F.; King, John A.

    1968-01-01

    The concept of home range was expressed by Seton (1909) in the term 'home region,' which Burr (1940, 1943) clarified with a definition of home range and exemplified in a definitive study of Peromyscus in the field. Burt pointed out the ever-changing characteristics of home-range area and the consequent absence of boundaries in the usual sense--a finding verified by investigators thereafter. In the studies summarized in this paper, sizes of home ranges of Peromyscus varied within two magnitudes, approximately from 0.1 acre to ten acres, in 34 studies conducted in a variety of habitats from the seaside dunes of Florida to the Alaskan forests. Variation in sizes of home ranges was correlated with both environmental and physiological factors; with habitat it was conspicuous, both in the same and different regions. Food supply also was related to size of home range, both seasonally and in relation to habitat. Home ranges generally were smallest in winter and largest in spring, at the onset of the breeding season. Activity and size also were affected by changes in weather. Activity was least when temperatures were low and nights were bright. Effects of rainfall were variable. Sizes varied according to sex and age; young mice remained in the parents' range until they approached maturity, when they began to travel more widely. Adult males commonly had larger home ranges than females, although there were a number of exceptions. An inverse relationship between population density and size of home range was shown in several studies and probably is the usual relationship. A basic need for activity and exploration also appeared to influence size of home range. Behavior within the home range was discussed in terms of travel patterns, travels in relation to home sites and refuges, territory, and stability of size of home range. Travels within the home range consisted of repeated use of well-worn trails to sites of food, shelter, and refuge, plus more random exploratory travels

  2. DOE Challenge Home Case Study: e2 Homes – Winter Park, Florida

    Energy Technology Data Exchange (ETDEWEB)

    none,

    2013-01-01

    This Challenge Home case study describes the first certified DOE Challenge Home as constructed by e2 Homes. Completed in May 2012, the “Wilson Residence” in Winter Park, Florida, is a 4,305-ft2 custom home that scores a HERS 57 without solar and a better than zero net-energy HERS -7 with solar.

  3. Recruitment and positioning determine the specific role of the XPF-ERCC1 endonuclease in interstrand crosslink repair.

    Science.gov (United States)

    Klein Douwel, Daisy; Hoogenboom, Wouter S; Boonen, Rick Acm; Knipscheer, Puck

    2017-07-14

    XPF-ERCC1 is a structure-specific endonuclease pivotal for several DNA repair pathways and, when mutated, can cause multiple diseases. Although the disease-specific mutations are thought to affect different DNA repair pathways, the molecular basis for this is unknown. Here we examine the function of XPF-ERCC1 in DNA interstrand crosslink (ICL) repair. We used Xenopus egg extracts to measure both ICL and nucleotide excision repair, and we identified mutations that are specifically defective in ICL repair. One of these separation-of-function mutations resides in the helicase-like domain of XPF and disrupts binding to SLX4 and recruitment to the ICL A small deletion in the same domain supports recruitment of XPF to the ICL, but inhibited the unhooking incisions most likely by disrupting a second, transient interaction with SLX4. Finally, mutation of residues in the nuclease domain did not affect localization of XPF-ERCC1 to the ICL but did prevent incisions on the ICL substrate. Our data support a model in which the ICL repair-specific function of XPF-ERCC1 is dependent on recruitment, positioning and substrate recognition. © 2017 The Authors. Published under the terms of the CC BY 4.0 license.

  4. A comparative study of cold- and warm-adapted Endonucleases A using sequence analyses and molecular dynamics simulations.

    Directory of Open Access Journals (Sweden)

    Davide Michetti

    Full Text Available The psychrophilic and mesophilic endonucleases A (EndA from Aliivibrio salmonicida (VsEndA and Vibrio cholera (VcEndA have been studied experimentally in terms of the biophysical properties related to thermal adaptation. The analyses of their static X-ray structures was no sufficient to rationalize the determinants of their adaptive traits at the molecular level. Thus, we used Molecular Dynamics (MD simulations to compare the two proteins and unveil their structural and dynamical differences. Our simulations did not show a substantial increase in flexibility in the cold-adapted variant on the nanosecond time scale. The only exception is a more rigid C-terminal region in VcEndA, which is ascribable to a cluster of electrostatic interactions and hydrogen bonds, as also supported by MD simulations of the VsEndA mutant variant where the cluster of interactions was introduced. Moreover, we identified three additional amino acidic substitutions through multiple sequence alignment and the analyses of MD-based protein structure networks. In particular, T120V occurs in the proximity of the catalytic residue H80 and alters the interaction with the residue Y43, which belongs to the second coordination sphere of the Mg2+ ion. This makes T120V an amenable candidate for future experimental mutagenesis.

  5. ConductHome: Gesture Interface Control of Home Automation Boxes

    OpenAIRE

    J. Branstett; V. Gagneux; A. Leleu; B. Levadoux; J. Pascale

    2015-01-01

    This paper presents the interface ConductHome which controls home automation systems with a Leap Motion using "invariant gesture protocols". This interface is meant to simplify the interaction of the user with its environment. A hardware part allows the Leap Motion to be carried around the house. A software part interacts with the home automation box and displays the useful information for the user. An objective of this work is the development of a natural/invariant/simpl...

  6. The contribution of DNA apurinic/apyrimidinic endonuclease genotype and smoking habit to Taiwan lung cancer risk.

    Science.gov (United States)

    Chen, Wei-Chun; Tsai, Chia-Wen; Hsia, Te-Chun; Chang, Wen-Shin; Lin, Liang-Yi; Liang, Shinn-Jye; Tu, Chih-Yen; Cheng, Wei-Erh; Chen, Hung-Jen; Wang, Shu-Ming; Bau, da-Tian

    2013-06-01

    To evaluate the association and interaction of genotypic polymorphism the gene for DNA-apurinic/apyrimidinic endonuclease (APEX1) with personal smoking habit and lung cancer risk in Taiwan, the polymorphic variants of APEX1, Asp(148)Glu (rs1130409), were analyzed in association with lung cancer risk, and their joint effect with personal smoking habits on lung cancer susceptibility was discussed. In this hospital-based case-control study, 358 patients with lung cancer and 716 cancer-free controls, frequency-matched by age and sex, were recruited and genotyped by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP). The results showed that the percentages of TT, TG and GG APEX1 Asp(148)Glu genotypes were not significantly different at 43.0%, 41.1% and 15.9% in the lung cancer patient group and 39.9%, 46.1% and 14.0% in non-cancer control group, respectively. We further analyzed the genetic-lifestyle effects on lung cancer risk and found the contribution of APEX1 Asp(148)Glu genotypes to lung cancer susceptibility was neither enhanced in the cigarette smokers nor in the non-smokers (p=0.3550 and 0.8019, respectively). Our results provide evidence that the non-synonymous polymorphism of APEX1 Asp(148)Glu may not be directly associated with lung cancer risk, nor enhance the effects of smoking habit on lung cancer development.

  7. Cost-effectiveness implications based on a comparison of nursing home and home health case mix.

    OpenAIRE

    Kramer, A M; Shaughnessy, P W; Pettigrew, M L

    1985-01-01

    Case-mix differences between 653 home health care patients and 650 nursing home patients, and between 455 Medicare home health patients and 447 Medicare nursing home patients were assessed using random samples selected from 20 home health agencies and 46 nursing homes in 12 states in 1982 and 1983. Home health patients were younger, had shorter lengths of stay, and were less functionally disabled than nursing home patients. Traditional long-term care problems requiring personal care were more...

  8. TRAVEL AND HOME LEAVE

    CERN Multimedia

    Human Resources Division

    2002-01-01

    Administrative procedures for : Travel to the home station and home leave (hl) Additional travel to the home station (at) Travel to the home station and home leave for family reasons (hlf) As part of the process of simplifying administrative procedures, HR and AS Divisions have devised a new, virtually automatic procedure for payment of travel expenses to the home station. The changes are aimed at rationalising administrative procedures and not at reducing benefits. The conditions of eligibility are unchanged. The new procedure, which will be operational with effect from 1st June 2002, will greatly simplify the administrative processing of claims for travel expenses and the recording of home leaves. Currently, requests for payment are introduced manually into the Advances and Claims system (AVCL) by divisional secretariats. All travel to the home station starting prior to 1st June 2002 will be processed according to the existing system whereas that starting on 1st June and after will be processed accordi...

  9. Dimensions of work-home culture and their relations with the use of work-home arrangements and work-home interaction

    NARCIS (Netherlands)

    Dikkers, J.S.E.; Geurts, S.A.E.; Dulk, L. den; Peper, A.B.; Taris, T.W.; Kompier, M.A.J.

    2007-01-01

    This study examined the associations of work–home culture with (a) demographic and organizational characteristics, (b) the use of work–home arrangements, and (c) negative and positive work–home interaction, among 1,179 employees from one public and two private organizations. Substantial support was

  10. Dimensions of work-home culture and their relations with the use of work-home arrangements and work-home interaction

    NARCIS (Netherlands)

    Dikkers, J.S.E.; Geurts, S.A.E.; den Dulk, L.; Peper, B.; Taris, T.W.; Kompier, M.A.J.

    2007-01-01

    This study examined the associations of work-home culture with (a) demographic and organizational characteristics, (b) the use of work-home arrangements, and (c) negative and positive work-home interaction, among 1,179 employees from one public and two private organizations. Substantial support was

  11. 6. Home deliveries

    African Journals Online (AJOL)

    Sitwala

    determine factors associated with home deliveries. Main outcome ... deliver at home than a health facility compared to those who .... regression analysis, women who had four years of schooling or .... by report bias, the burden of home deliveries is a real challenge .... Journal of Econometrics 1987; 36: 185-204. 14. Michelo ...

  12. Home Energy Saver

    Science.gov (United States)

    release announcing Home Energy Saver and a Q-and-A. The "About" page should tell you everything you need to know about using Home Energy Saver. If you have any questions, please email the project leader, Dr. Evan Mills. News Releases Microsoft Licenses Berkeley Lab's Home Energy Saver Code for Its

  13. DOE Zero Energy Ready Home Case Study: Garbett Homes, Herriman, Utah

    Energy Technology Data Exchange (ETDEWEB)

    none,

    2013-09-01

    As the first net zero-energy production home certified in Utah, this house incorporates two 94% efficient tankless water heaters and two roof-mounted solar panels that preheat the home's water supply. This home won a 2013 Housing Innovation Award in the production builder category.

  14. Work-home interference in relation to work, organizational, and home characteristics

    NARCIS (Netherlands)

    Dikkers, J.S.E.

    2008-01-01

    In this thesis we examine work-home interference in relation to work, organizational, and home characteristics. The study is guided by the following three research questions: i) how are workload and negative WHI temporally related? (research question 1); ii) is work-home culture related to the use

  15. Does hospital at home for palliative care facilitate death at home? Randomised controlled trial

    Science.gov (United States)

    Grande, Gunn E; Todd, Chris J; Barclay, Stephen I G; Farquhar, Morag C

    1999-01-01

    Objective To evaluate the impact on place of death of a hospital at home service for palliative care. Design Pragmatic randomised controlled trial. Setting Former Cambridge health district. Participants 229 patients referred to the hospital at home service; 43 randomised to control group (standard care), 186 randomised to hospital at home. Intervention Hospital at home versus standard care. Main outcome measures Place of death. Results Twenty five (58%) control patients died at home compared with 124 (67%) patients allocated to hospital at home. This difference was not significant; intention to treat analysis did not show that hospital at home increased the number of deaths at home. Seventy three patients randomised to hospital at home were not admitted to the service. Patients admitted to hospital at home were significantly more likely to die at home (88/113; 78%) than control patients. It is not possible to determine whether this was due to hospital at home itself or other characteristics of the patients admitted to the service. The study attained less statistical power than initially planned. Conclusion In a locality with good provision of standard community care we could not show that hospital at home allowed more patients to die at home, although neither does the study refute this. Problems relating to recruitment, attrition, and the vulnerability of the patient group make randomised controlled trials in palliative care difficult. While these difficulties have to be recognised they are not insurmountable with the appropriate resourcing and setting. Key messagesTerminally ill patients allocated to hospital at home were no more likely to die at home than patients receiving standard careAlthough the subsample of patients actually admitted to hospital at home did show a significant increase in likelihood of dying at home, whether this was due to the service itself or the characteristics of patients admitted to hospital at home could not be determinedThe need to

  16. Cell-autonomous progeroid changes in conditional mouse models for repair endonuclease XPG deficiency.

    Directory of Open Access Journals (Sweden)

    Sander Barnhoorn

    2014-10-01

    Full Text Available As part of the Nucleotide Excision Repair (NER process, the endonuclease XPG is involved in repair of helix-distorting DNA lesions, but the protein has also been implicated in several other DNA repair systems, complicating genotype-phenotype relationship in XPG patients. Defects in XPG can cause either the cancer-prone condition xeroderma pigmentosum (XP alone, or XP combined with the severe neurodevelopmental disorder Cockayne Syndrome (CS, or the infantile lethal cerebro-oculo-facio-skeletal (COFS syndrome, characterized by dramatic growth failure, progressive neurodevelopmental abnormalities and greatly reduced life expectancy. Here, we present a novel (conditional Xpg-/- mouse model which -in a C57BL6/FVB F1 hybrid genetic background- displays many progeroid features, including cessation of growth, loss of subcutaneous fat, kyphosis, osteoporosis, retinal photoreceptor loss, liver aging, extensive neurodegeneration, and a short lifespan of 4-5 months. We show that deletion of XPG specifically in the liver reproduces the progeroid features in the liver, yet abolishes the effect on growth or lifespan. In addition, specific XPG deletion in neurons and glia of the forebrain creates a progressive neurodegenerative phenotype that shows many characteristics of human XPG deficiency. Our findings therefore exclude that both the liver as well as the neurological phenotype are a secondary consequence of derailment in other cell types, organs or tissues (e.g. vascular abnormalities and support a cell-autonomous origin caused by the DNA repair defect itself. In addition they allow the dissection of the complex aging process in tissue- and cell-type-specific components. Moreover, our data highlight the critical importance of genetic background in mouse aging studies, establish the Xpg-/- mouse as a valid model for the severe form of human XPG patients and segmental accelerated aging, and strengthen the link between DNA damage and aging.

  17. Second-home electricity consumption

    International Nuclear Information System (INIS)

    Andersen, Frits M.; Christensen, Morten S.; Jensen, Ole Michael; Kofoed, Niels-Ulrik; Morthorst, Poul Erik

    2008-01-01

    In Denmark, electricity consumption in first and second homes has developed quite differently. Since 1990, electricity consumption in ordinary residences has grown moderately, while consumption in weekend and second homes has increased considerably. In turn, this development has been blamed on a growing number of luxury cottages, new legislation permitting senior citizens to have their permanent address in their second home and a growing number of electric appliances. In order to examine the growing electricity consumption in second homes and to estimate future demand, a multidisciplinary study combining top-down and bottom-up analyses was conducted, i.e., combining models using aggregated economic parameters and feasibility studies using technical parameters, respectively. The top-down estimation showed that changes in electricity consumption in second homes correlate to changes in income. The bottom-up estimation showed that consumption was mainly affected by the frequency with which second homes were used in the winter time. This indicates that additional second homes, increased full-time use and intensified use of electric appliances are the main reasons for the observed increases in electricity consumption. Luxury tourism use and senior citizens' that use a few per cent of the second homes as their home contribute to a minor degree to the overall increase of electricity consumption. Scenarios show that this development may accelerate with increased leisure time, increased use and more permanent settlement in second homes

  18. Leaving home in Denmark

    DEFF Research Database (Denmark)

    Nielsen, Rikke Skovgaard

    2015-01-01

    The paper focuses on ethnic differences in the timing and patterns of leaving the parental home. Leaving home is a key transition in the life course of the individual, and extensive research has been conducted on the timing and patterns of leaving it. However, ethnic differences in these patterns...... of leaving home. Results showed that while some differences disappeared when controlling for covariates, others persisted, thus indicating ethnic differences in home-leaving patterns. A strong link between leaving home and marriage was substantiated for Turks, but not for Somalis. The home-leaving patterns...... of Somalis were much more similar to those of Danes. Overall, Turkish descendants were similar to Turkish immigrants but with some differentiation. The analyses identified the existence of ethnic differences in home-leaving patterns but also found evidence of a shift towards less traditional patterns, i...

  19. Magnonic Crystal as a Delay Line for Low-Noise Auto-Oscillator

    Science.gov (United States)

    2015-05-12

    Magnonic crystal as a delay line for low-noise auto-oscillator Elena Bankowski and Thomas Meitzler U.S. Army TARDEC, Warren, Michigan 48397, USA...authors propose to use the magnonic crystal patterned on the YIG magnetic film as an efficient delay line in the feedback loop of tunable auto-oscillator...increasing the thickness of such delay line as compare to the YIG film with no pattern. In turn, use of this magnonic crystal opens a way to improve

  20. Giant Faraday Rotation in Metal-Fluoride Nanogranular Films.

    Science.gov (United States)

    Kobayashi, N; Ikeda, K; Gu, Bo; Takahashi, S; Masumoto, H; Maekawa, S

    2018-03-21

    Magneto-optical Faraday effect is widely applied in optical devices and is indispensable for optical communications and advanced information technology. However, the bismuth garnet Bi-YIG is only the Faraday material since 1972. Here we introduce (Fe, FeCo)-(Al-,Y-fluoride) nanogranular films exhibiting giant Faraday effect, 40 times larger than Bi-YIG. These films have a nanocomposite structure, in which nanometer-sized Fe, FeCo ferromagnetic granules are dispersed in a Al,Y-fluoride matrix.

  1. Beyond Home Automation: Designing More Effective Smart Home Systems

    OpenAIRE

    Carner, Paolo

    2009-01-01

    This paper outlines a Smart Home Proof-of-Concept system that uses a Bayesian Network to predict the likelihood of a monitored event to occur. Firstly, this paper will provide an introduction to the concept of a smart home system; then it will outline how Artificial Intelligence concepts can be used to make such systems more effective. Finally, it will detail the implementation of a smart home system, which uses an inference engine to determine the likelihood of a fire. The system prototype h...

  2. Smart homes, private homes? An empirical study of technology researchers? perceptions of ethical issues in developing smart-home health technologies

    OpenAIRE

    Birchley, Giles; Huxtable, Richard; Murtagh, Madeleine; ter Meulen, Ruud; Flach, Peter; Gooberman-Hill, Rachael

    2017-01-01

    Background Smart-home technologies, comprising environmental sensors, wearables and video are attracting interest in home healthcare delivery. Development of such technology is usually justified on the basis of the technology?s potential to increase the autonomy of people living with long-term conditions. Studies of the ethics of smart-homes raise concerns about privacy, consent, social isolation and equity of access. Few studies have investigated the ethical perspectives of smart-home engine...

  3. At Home in the Office. A Guide for the Home Worker.

    Science.gov (United States)

    Thomas, Neil; And Others

    This guide provides information to persons interested in establishing a work-at-home program, specifically those in clerical or support staff positions who use modern automated office equipment. The text is divided into two sections. The Home Worker section provides a summary of the At Home in the Office Project, personality characteristics…

  4. Evolution of Home Automation Technology

    OpenAIRE

    Mohd. Rihan; M. Salim Beg

    2009-01-01

    In modern society home and office automation has becomeincreasingly important, providing ways to interconnectvarious home appliances. This interconnection results infaster transfer of information within home/offices leading tobetter home management and improved user experience.Home Automation, in essence, is a technology thatintegrates various electrical systems of a home to provideenhanced comfort and security. Users are grantedconvenient and complete control over all the electrical homeappl...

  5. More Than Four Walls: The Meaning of Home in Home Birth Experiences

    Directory of Open Access Journals (Sweden)

    Emily Burns

    2015-04-01

    Full Text Available The “home versus hospital” as places of birth debate has had a long and at times vicious history. From academic literature to media coverage, the two have often been pitted against each other not only as opposing physical spaces, but also as opposing ideologies of birth. The hospital has been heavily critiqued as a site of childbirth since the 1960s, with particular focus on childbirth and medicalisation. The focus of much of the hospital and home birthing research exists on a continuum of medicalisation, safety, risk, agency, and maternal and neonatal health and wellbeing. While the hospital birthing space has been interrogated, a critique of home birthing space has remained largely absent from the social sciences. The research presented in this article unpacks the complex relationship between home birthing women and the spaces in which they birth. Using qualitative data collected with 59 home birthing women in Australia in 2010, between childbearing and the home should not be considered as merely an alternative to hospital births, but rather as an experience that completely renegotiates the home space. Home, for the participants in this study, is a dynamic, changing, and even spiritual element in the childbirth experience, and not simply the building in which it occurs.

  6. Second-home electricity consumption

    Energy Technology Data Exchange (ETDEWEB)

    Andersen, Frits M.; Morthorst, Poul Erik [Risoe, Systems Analysis Department, Technical University of Denmark, P.O. Box 49, DK-4000 Roskilde (Denmark); Christensen, Morten S.; Kofoed, Niels-Ulrik [Esbensen Consulting Engineers, Carl Jacobsens Vej 25D, DK-2500 Valby (Denmark); Jensen, Ole Michael [Danish Building Research Institute, Department of Energy and Environment, University of Aalborg, P.O. Box 119, DK-2970 Horsholm (Denmark)

    2008-01-15

    In Denmark, electricity consumption in first and second homes has developed quite differently. Since 1990, electricity consumption in ordinary residences has grown moderately, while consumption in weekend and second homes has increased considerably. In turn, this development has been blamed on a growing number of luxury cottages, new legislation permitting senior citizens to have their permanent address in their second home and a growing number of electric appliances. In order to examine the growing electricity consumption in second homes and to estimate future demand, a multidisciplinary study combining top-down and bottom-up analyses was conducted, i.e., combining models using aggregated economic parameters and feasibility studies using technical parameters, respectively. The top-down estimation showed that changes in electricity consumption in second homes correlate to changes in income. The bottom-up estimation showed that consumption was mainly affected by the frequency with which second homes were used in the winter time. This indicates that additional second homes, increased full-time use and intensified use of electric appliances are the main reasons for the observed increases in electricity consumption. Luxury tourism use and senior citizens' that use a few per cent of the second homes as their home contribute to a minor degree to the overall increase of electricity consumption. Scenarios show that this development may accelerate with increased leisure time, increased use and more permanent settlement in second homes. (author)

  7. DOE Zero Energy Ready Home Case Study: Amaris Homes, Afton Model

    Energy Technology Data Exchange (ETDEWEB)

    Pacific Northwest National Laboratory

    2017-09-01

    Amaris Homes built this 3,734-ft2 home in Afton, Minnesota, to the performance criteria of the DOE Zero Energy Ready Home (ZERH) program. A high-efficiency gas boiler provides hot water for the zoned radiant floor system as well as for faucets and showers. A high-efficiency heat pump provides zoned cooling.

  8. Strategy Guideline: Demonstration Home

    Energy Technology Data Exchange (ETDEWEB)

    Savage, C.; Hunt, A.

    2012-12-01

    This guideline will provide a general overview of the different kinds of demonstration home projects, a basic understanding of the different roles and responsibilities involved in the successful completion of a demonstration home, and an introduction into some of the lessons learned from actual demonstration home projects. Also, this guideline will specifically look at the communication methods employed during demonstration home projects. And lastly, we will focus on how to best create a communication plan for including an energy efficient message in a demonstration home project and carry that message to successful completion.

  9. Strategy Guideline. Demonstration Home

    Energy Technology Data Exchange (ETDEWEB)

    Hunt, A.; Savage, C.

    2012-12-01

    This guideline will provide a general overview of the different kinds of demonstration home projects, a basic understanding of the different roles and responsibilities involved in the successful completion of a demonstration home, and an introduction into some of the lessons learned from actual demonstration home projects. Also, this guideline will specifically look at the communication methods employed during demonstration home projects. And lastly, we will focus on how to best create a communication plan for including an energy efficient message in a demonstration home project and carry that message to successful completion.

  10. A comparison of home care clients and nursing home residents: can community based care keep the elderly and disabled at home?

    Science.gov (United States)

    Shugarman, L R; Fries, B E; James, M

    1999-01-01

    Admission cohorts from the Michigan Medicaid Home and Community-Based Waiver program and Ohio nursing homes were compared on measures of resource utilization including a modified Resource Utilization Groups (RUG-III) system, Activities of Daily Living (ADLs), and overall case mix. We found that, contrary to previous research, the two samples were remarkably similar across RUG-III categories. However, the nursing home sample was more functionally impaired on measures of ADL functioning and overall case mix. Results of this study may inform policymakers and providers of the potential for maintaining the appropriate population in the home with government-funded home care.

  11. At-home and away-from-home dietary patterns and BMI z-scores in Brazilian adolescents.

    Science.gov (United States)

    Cunha, Diana Barbosa; Bezerra, Ilana Nogueira; Pereira, Rosangela Alves; Sichieri, Rosely

    2018-01-01

    Away-from-home food intake has been associated with high rates of overweight among children and adolescents. However, there are no studies comparing at-home and away-from-home eating patterns among adolescents. The objective of this paper was to identify at-home and away-from-home dietary patterns among adolescents in Brazil, and to evaluate the relationship between these patterns and body mass index (BMI) z-scores. Data from the Brazilian National Dietary Survey 2008-2009 were analyzed in this cross-sectional study. Dietary intake was assessed by completion of written food records on two non-consecutive days. Five thousand two hundred sixty-six adolescents 10-19 years of age living in urban areas of Brazil were included in the analysis. Thirty-two food groups were examined by factor analysis, stratified by at-home and away-from-home eating. The associations between the food patterns and BMI z-scores were ascertained using linear regression analysis. In general, mean at-home food intake was greater than away-from-home food intake, but the ratio of away-from-home/at-home was greater than 30% for baked and deep-fried snacks, soft drinks, sandwiches, pizza, and desserts, and was lower than 10% for rice and beans. Three main similar dietary patterns were identified both at-home and away-from-home: the "Traditional pattern", the "Bread and Butter pattern" and the "Western pattern"; however, away-from-home patterns encompassed more overall food items. Only the at-home "Western pattern" was positively associated with BMI z-scores (β = 0.0006; p away-from-home food consumption is not associated. Copyright © 2017 Elsevier Ltd. All rights reserved.

  12. Alert management for home healthcare based on home automation analysis.

    Science.gov (United States)

    Truong, T T; de Lamotte, F; Diguet, J-Ph; Said-Hocine, F

    2010-01-01

    Rising healthcare for elder and disabled people can be controlled by offering people autonomy at home by means of information technology. In this paper, we present an original and sensorless alert management solution which performs multimedia and home automation service discrimination and extracts highly regular home activities as sensors for alert management. The results of simulation data, based on real context, allow us to evaluate our approach before application to real data.

  13. Altered endoribonuclease activity of apurinic/apyrimidinic endonuclease 1 variants identified in the human population.

    Directory of Open Access Journals (Sweden)

    Wan Cheol Kim

    Full Text Available Apurinic/apyrimidinic endonuclease 1 (APE1 is the major mammalian enzyme in the DNA base excision repair pathway and cleaves the DNA phosphodiester backbone immediately 5' to abasic sites. APE1 also has 3'-5' DNA exonuclease and 3' DNA phosphodiesterase activities, and regulates transcription factor DNA binding through its redox regulatory function. The human APE1 has recently been shown to endonucleolytically cleave single-stranded regions of RNA. Towards understanding the biological significance of the endoribonuclease activity of APE1, we examined eight different amino acid substitution variants of APE1 previously identified in the human population. Our study shows that six APE1 variants, D148E, Q51H, I64V, G241R, R237A, and G306A, exhibit a 76-85% reduction in endoribonuclease activity against a specific coding region of the c-myc RNA, yet fully retain the ability to cleave apurinic/apyrimidinic DNA. We found that two APE1 variants, L104R and E126D, exhibit a unique RNase inhibitor-resistant endoribonuclease activity, where the proteins cleave c-myc RNA 3' of specific single-stranded guanosine residues. Expression of L104R and E126D APE1 variants in bacterial Origami cells leads to a 60-80% reduction in colony formation and a 1.5-fold increase in cell doubling time, whereas the other variants, which exhibit diminished endoribonuclease activity, had no effect. These data indicate that two human APE1 variants exhibit a unique endoribonuclease activity, which correlates with their ability to induce cytotoxicity or slow down growth in bacterial cells and supports the notion of their biological functionality.

  14. 16 CFR 460.16 - What new home sellers must tell new home buyers.

    Science.gov (United States)

    2010-01-01

    ... LABELING AND ADVERTISING OF HOME INSULATION § 460.16 What new home sellers must tell new home buyers. If you are a new home seller, you must put the following information in every sales contract: The type... exception to this rule. If the buyer signs a sales contract before you know what type of insulation will be...

  15. [Home births].

    Science.gov (United States)

    Welffens, K; Kirkpatrick, C; Daelemans, C; Derisbourg, S

    In Belgium, very few women give birth outside the delivery room. In the United Kingdom and in the Netherlands, they are more numerous. Several studies evaluated obstetric and neonatal outcomes of home births compared with hospital births. We selected seven recent and large studies (with cohorts of more than 5.000 women) using PubMed, Science Direct and Cochrane Database of Systematic Reviews. Several questions were examined. Is there any difference in maternal and neonatal outcomes depending on the intended place of birth? Does parity affect outcomes ? What are the characteristics of women who choose to deliver at home ? We conclude that giving birth at home improves obstetric outcomes but is riskier for the baby, especially for the first one. The women delivering at home are mainly white Europeans, between 25 and 35 years old, in a relationship, multiparous and wealthier. In order to avoid this increased risk for the baby while preserving the obstetric advantages, alongside birth centers offer an intermediate solution. They combine the reassuring home-like atmosphere with the safety of the hospital. In Belgium, the first alongside birth center " Le Cocon " (a low technicity unit distinct from the delivery room) offers now this type of alternative place of birth for women in Hôpital Erasme in Brussels.

  16. SETI@home

    Science.gov (United States)

    Project Help Donate Porting Graphics Add-ons Science About SETI@home About Astropulse Science Community Message boards Questions and Answers Teams Profiles User search Web sites Pictures and music User University of California SETI@home and Astropulse are funded by grants from the National Science Foundation

  17. Paramedic-Initiated Home Care Referrals and Use of Home Care and Emergency Medical Services.

    Science.gov (United States)

    Verma, Amol A; Klich, John; Thurston, Adam; Scantlebury, Jordan; Kiss, Alex; Seddon, Gayle; Sinha, Samir K

    2018-01-01

    We examined the association between paramedic-initiated home care referrals and utilization of home care, 9-1-1, and Emergency Department (ED) services. This was a retrospective cohort study of individuals who received a paramedic-initiated home care referral after a 9-1-1 call between January 1, 2011 and December 31, 2012 in Toronto, Ontario, Canada. Home care, 9-1-1, and ED utilization were compared in the 6 months before and after home care referral. Nonparametric longitudinal regression was performed to assess changes in hours of home care service use and zero-inflated Poisson regression was performed to assess changes in the number of 9-1-1 calls and ambulance transports to ED. During the 24-month study period, 2,382 individuals received a paramedic-initiated home care referral. After excluding individuals who died, were hospitalized, or were admitted to a nursing home, the final study cohort was 1,851. The proportion of the study population receiving home care services increased from 18.2% to 42.5% after referral, representing 450 additional people receiving services. In longitudinal regression analysis, there was an increase of 17.4 hours in total services per person in the six months after referral (95% CI: 1.7-33.1, p = 0.03). The mean number of 9-1-1 calls per person was 1.44 (SD 9.58) before home care referral and 1.20 (SD 7.04) after home care referral in the overall study cohort. This represented a 10% reduction in 9-1-1 calls (95% CI: 7-13%, p home care referral and 0.79 (SD 6.27) after home care referral, representing a 7% reduction (95% CI: 3-11%, p home care records were included in the analysis, the reductions in 9-1-1 calls and ambulance transports to ED were attenuated but remained statistically significant. Paramedic-initiated home care referrals in Toronto were associated with improved access to and use of home care services and may have been associated with reduced 9-1-1 calls and ambulance transports to ED.

  18. New American Home(regsign): Atlanta, Georgia - 2001; Building America--The New American Home

    International Nuclear Information System (INIS)

    IBACOS; Anderson, R.

    2001-01-01

    The New American Home(reg s ign) is an annual showcase project designed by committee and co-sponsored by the National Association of Home Builders' National Council of the Housing Industry, BUILDERS Magazine, and Ladies Home Journal. Hedgewood Properties teamed with Building America's IBACOS Consortium and Southface Energy Institute to build a house with a Home Energy Rating Systems (HERS) level of 90

  19. Digital Living at Home

    DEFF Research Database (Denmark)

    Andersen, Pernille Viktoria Kathja; Christiansen, Ellen Tove

    2013-01-01

    of these user voices has directed us towards a ‘home-keeping’ design discourse, which opens new horizons for design of digital home control systems by allowing users to perform as self-determined controllers and groomers of their habitat. The paper concludes by outlining the implications of a ‘home......Does living with digital technology inevitably lead to digital living? Users talking about a digital home control system, they have had in their homes for eight years, indicate that there is more to living with digital technology than a functional-operational grip on regulation. Our analysis......-keeping’ design discourse....

  20. Respiratory Home Health Care

    Science.gov (United States)

    ... Us Home > Healthy Living > Living With Lung Disease > Respiratory Home Health Care Font: Aerosol Delivery Oxygen Resources ... Teenagers Living With Lung Disease Articles written by Respiratory Experts Respiratory Home Health Care Respiratory care at ...

  1. Electrical properties of single crystal Yttrium Iron Garnet ultra-thin films at high temperatures

    OpenAIRE

    Thiery, Nicolas; Naletov, Vladimir V.; Vila, Laurent; Marty, Alain; Brenac, Ariel; Jacquot, Jean-François; de Loubens, Grégoire; Viret, Michel; Anane, Abdelmadjid; Cros, Vincent; Youssef, Jamal Ben; Demidov, Vladislav E.; Demokritov, Sergej O.; Klein, Olivier

    2017-01-01

    We report a study on the electrical properties of 19 nm thick Yttrium Iron Garnet (YIG) films grown by liquid phase epitaxy. The electrical conductivity and Hall coefficient are measured in the high temperature range [300,400]~K using a Van der Pauw four-point probe technique. We find that the electrical resistivity decreases exponentially with increasing temperature following an activated behavior corresponding to a band-gap of $E_g\\approx 2$ eV, indicating that epitaxial YIG ultra-thin film...

  2. Asthma Home Environment Checklist

    Science.gov (United States)

    This checklist guides home care visitors in identifying environmental asthma triggers most commonly found in homes. It includes sections on the building, home interior and room interior and provides low-cost action steps for remediation.

  3. DOE Zero Energy Ready Home Case Study: One Sky Homes — Cottle Zero Net Energy Home, San Jose, CA

    Energy Technology Data Exchange (ETDEWEB)

    none,

    2014-09-01

    This builder took home the Grand Winner prize in the Custom Builder category in the 2014 Housing Innovation Awards for its high performance building science approach. The builder used insulated concrete form blocks to create the insulated crawlspace foundation for its first DOE Zero Energy Ready Home, the first net zero energy new home certified in the state of California.

  4. "Home is always home" : (former) street youth in Blantyre, Malawi, and the fluidity of constructing home

    NARCIS (Netherlands)

    Hendriks, T.D.

    2016-01-01

    For many Malawians the concept of home is strongly associated with the rural areas and one's (supposedly rural) place of birth. This 'grand narrative about home', though often reiterated, doesn't necessarily depict lived reality. Malawi's history of movement and labor migration coupled with

  5. "Home is always home" : (former) street youth in Blantyre, Malawi, and the fluidity of constructing home

    NARCIS (Netherlands)

    Hendriks, T.D.

    2017-01-01

    For many Malawians the concept of home is strongly associated with the rural areas and one's (supposedly rural) place of birth. This 'grand narrative about home', though often reiterated, doesn't necessarily depict lived reality. Malawi's history of movement and labor migration coupled with

  6. Smart homes, private homes? An empirical study of technology researchers' perceptions of ethical issues in developing smart-home health technologies.

    Science.gov (United States)

    Birchley, Giles; Huxtable, Richard; Murtagh, Madeleine; Ter Meulen, Ruud; Flach, Peter; Gooberman-Hill, Rachael

    2017-04-04

    Smart-home technologies, comprising environmental sensors, wearables and video are attracting interest in home healthcare delivery. Development of such technology is usually justified on the basis of the technology's potential to increase the autonomy of people living with long-term conditions. Studies of the ethics of smart-homes raise concerns about privacy, consent, social isolation and equity of access. Few studies have investigated the ethical perspectives of smart-home engineers themselves. By exploring the views of engineering researchers in a large smart-home project, we sought to contribute to dialogue between ethics and the engineering community. Either face-to-face or using Skype, we conducted in-depth qualitative interviews with 20 early- and mid-career smart-home researchers from a multi-centre smart-home project, who were asked to describe their own experience and to reflect more broadly about ethical considerations that relate to smart-home design. With participants' consent, interviews were audio-recorded, transcribed and analysed using a thematic approach. Two overarching themes emerged: in 'Privacy', researchers indicated that they paid close attention to negative consequences of potential unauthorised information sharing in their current work. However, when discussing broader issues in smart-home design beyond the confines of their immediate project, researchers considered physical privacy to a lesser extent, even though physical privacy may manifest in emotive concerns about being watched or monitored. In 'Choice', researchers indicated they often saw provision of choice to end-users as a solution to ethical dilemmas. While researchers indicated that choices of end-users may need to be restricted for technological reasons, ethical standpoints that restrict choice were usually assumed and embedded in design. The tractability of informational privacy may explain the greater attention that is paid to it. However, concerns about physical privacy may

  7. Telemedicine in Neonatal Home Care

    DEFF Research Database (Denmark)

    Holm, Kristina Garne; Brødsgaard, Anne; Zachariassen, Gitte

    2016-01-01

    participatory design and qualitative methods. Data were collected from observational studies, individual interviews, and focus group interviews. Two neonatal units participated. One unit was experienced in providing neonatal home care with home visits, and the other planned to offer neonatal home care......BACKGROUND: For the majority of preterm infants, the last weeks of hospital admission mainly concerns tube feeding and establishment of breastfeeding. Neonatal home care (NH) was developed to allow infants to remain at home for tube feeding and establishment of breastfeeding with regular home...... visits from neonatal nurses. For hospitals covering large regions, home visits may be challenging, time consuming, and expensive and alternative approaches must be explored. OBJECTIVE: To identify parental needs when wanting to provide neonatal home care supported by telemedicine. METHODS: The study used...

  8. Making the Transition from Traditional to Home Schooling: Home School Family Motivations

    Science.gov (United States)

    Anthony, Kenneth Vance; Burroughs, Susie

    2010-01-01

    This study examined the motivations of families that operate home schools. Four intact, religiously conservative families were interviewed and observed over one year. Findings showed that families were motivated by multiple factors to leave traditional schooling and begin home schooling. Additionally, the motivations to home school influenced the…

  9. Magnetic-field-induced decrease of the spin Peltier effect in Pt/Y3Fe5O12 system at room temperature

    Science.gov (United States)

    Itoh, Ryuichi; Iguchi, Ryo; Daimon, Shunsuke; Oyanagi, Koichi; Uchida, Ken-ichi; Saitoh, Eiji

    2017-11-01

    We report the observation of magnetic-field-induced decrease of the spin Peltier effect (SPE) in a junction of a paramagnetic metal Pt and a ferrimagnetic insulator Y3Fe5O12 (YIG) at room temperature. For driving the SPE, spin currents are generated via the spin Hall effect from applied charge currents in the Pt layer, and injected into the adjacent thick YIG film. The resultant temperature modulation is detected by a commonly used thermocouple attached to the Pt/YIG junction. The output of the thermocouple shows sign reversal when the magnetization is reversed and linearly increases with the applied current, demonstrating the detection of the SPE signal. We found that the SPE signal decreases with the magnetic field. The observed decreasing rate was found to be comparable to that of the spin Seebeck effect (SSE), suggesting the dominant and similar contribution of the low-energy magnons in the SPE as in the SSE.

  10. Acoustic spin pumping in magnetoelectric bulk acoustic wave resonator

    Directory of Open Access Journals (Sweden)

    N. I. Polzikova

    2016-05-01

    Full Text Available We present the generation and detection of spin currents by using magnetoelastic resonance excitation in a magnetoelectric composite high overtone bulk acoustic wave (BAW resonator (HBAR formed by a Al-ZnO-Al-GGG-YIG-Pt structure. Transversal BAW drives magnetization oscillations in YIG film at a given resonant magnetic field, and the resonant magneto-elastic coupling establishes the spin-current generation at the Pt/YIG interface. Due to the inverse spin Hall effect (ISHE this BAW-driven spin current is converted to a dc voltage in the Pt layer. The dependence of the measured voltage both on magnetic field and frequency has a resonant character. The voltage is determined by the acoustic power in HBAR and changes its sign upon magnetic field reversal. We compare the experimentally observed amplitudes of the ISHE electrical field achieved by our method and other approaches to spin current generation that use surface acoustic waves and microwave resonators for ferromagnetic resonance excitation, with the theoretically expected values.

  11. Frequency and magnetic field mapping of magnetoelastic spin pumping in high overtone bulk acoustic wave resonator

    Science.gov (United States)

    Polzikova, N. I.; Alekseev, S. G.; Pyataikin, I. I.; Luzanov, V. A.; Raevskiy, A. O.; Kotov, V. A.

    2018-05-01

    We report on the first observation of microvolt-scale inverse spin Hall effect (ISHE) dc voltage driven by an acoustic spin pumping (ASP) in a bulk acoustic wave (BAW) resonator formed by a Al-ZnO-Al-YIG(1)-GGG-YIG(2)-Pt structure. When 2 mW power is applied to an Al-ZnO-Al transducer, the voltage VISHE ˜ 4 μV in the Pt film is observed as a result of resonant ASP from YIG(2) to Pt in the area ˜ 170 μm. The results of frequency and magnetic field mapping of VISHE(f,H) together with reflectivity of the resonator show an obvious agreement between the positions of the voltage maxima and BAW resonance frequencies fn(H) on the (f, H) plane. At the same time a significant asymmetry of the VISHE(fn(H)) value in reference to the magnetoelastic resonance (MER) line fMER(H) position is revealed, which is explained by asymmetry of the magnetoelastic waves dispersion law.

  12. Efficient spin transport through polyaniline

    Science.gov (United States)

    Mendes, J. B. S.; Alves Santos, O.; Gomes, J. P.; Assis, H. S.; Felix, J. F.; Rodríguez-Suárez, R. L.; Rezende, S. M.; Azevedo, A.

    2017-01-01

    By using the spin pumping process, we show that it is possible to transport a pure spin current across layers of conducting polyaniline (PANI) with several hundred nanometers sandwiched between a film of the ferrimagnetic insulator yttrium iron garnet (YIG) and a thin layer of platinum. The spin current generated by microwave-driven ferromagnetic resonance of the YIG film, injected through the YIG/PANI interface, crosses the whole PANI layer and then is injected into the Pt layer. By means of the inverse spin Hall effect in the Pt, the spin current is converted into charge current and electrically detected as a dc voltage. We measured a spin diffusion length in PANI of 590 ± 40 nm, which is very large compared with normal metals, demonstrating that PANI can be used as an efficient spin current conductor and poor charge current conductor, opening the path towards spintronics applications based in this very attractive material.

  13. Optical Manipulation of a Magnon-Photon Hybrid System.

    Science.gov (United States)

    Braggio, C; Carugno, G; Guarise, M; Ortolan, A; Ruoso, G

    2017-03-10

    We demonstrate an all-optical method for manipulating the magnetization in a 1-mm yttrium-iron-garnet (YIG) sphere placed in a ∼0.17  T uniform magnetic field. A harmonic of the frequency comb delivered by a multi-GHz infrared laser source is tuned to the Larmor frequency of the YIG sphere to drive magnetization oscillations, which in turn give rise to a radiation field used to thoroughly investigate the phenomenon. The radiation damping issue that occurs at high frequency and in the presence of highly magnetizated materials has been overcome by exploiting the magnon-photon strong coupling regime in microwave cavities. Our findings demonstrate an effective technique for ultrafast control of the magnetization vector in optomagnetic materials via polarization rotation and intensity modulation of an incident laser beam. We eventually get a second-order susceptibility value of ∼10^{-7}  cm^{2}/MW for single crystal YIG.

  14. Acoustic spin pumping in magnetoelectric bulk acoustic wave resonator

    Energy Technology Data Exchange (ETDEWEB)

    Polzikova, N. I., E-mail: polz@cplire.ru; Alekseev, S. G.; Pyataikin, I. I.; Kotelyanskii, I. M.; Luzanov, V. A.; Orlov, A. P. [Kotel’nikov Institute of Radio Engineering and Electronics of Russian Academy of Sciences, Mokhovaya 11, building 7, Moscow, 125009 (Russian Federation)

    2016-05-15

    We present the generation and detection of spin currents by using magnetoelastic resonance excitation in a magnetoelectric composite high overtone bulk acoustic wave (BAW) resonator (HBAR) formed by a Al-ZnO-Al-GGG-YIG-Pt structure. Transversal BAW drives magnetization oscillations in YIG film at a given resonant magnetic field, and the resonant magneto-elastic coupling establishes the spin-current generation at the Pt/YIG interface. Due to the inverse spin Hall effect (ISHE) this BAW-driven spin current is converted to a dc voltage in the Pt layer. The dependence of the measured voltage both on magnetic field and frequency has a resonant character. The voltage is determined by the acoustic power in HBAR and changes its sign upon magnetic field reversal. We compare the experimentally observed amplitudes of the ISHE electrical field achieved by our method and other approaches to spin current generation that use surface acoustic waves and microwave resonators for ferromagnetic resonance excitation, with the theoretically expected values.

  15. A parasitic selfish gene that affects host promiscuity

    OpenAIRE

    Giraldo-Perez, Paulina; Goddard, Matthew R.

    2013-01-01

    Selfish genes demonstrate transmission bias and invade sexual populations despite conferring no benefit to their hosts. While the molecular genetics and evolutionary dynamics of selfish genes are reasonably well characterized, their effects on hosts are not. Homing endonuclease genes (HEGs) are one well-studied family of selfish genes that are assumed to be benign. However, we show that carrying HEGs is costly for Saccharomyces cerevisiae, demonstrating that these genetic elements are not nec...

  16. Ostomy Home Skills Program

    Medline Plus

    Full Text Available ... Ostomy Home Skills Program Ostomy Home Skills Program Adult Ostomy Pediatric Ostomy Programa de Destrezas para manejo Doméstico de Ostomía Ostomy Home Skills Program Adult Ostomy Pediatric Ostomy Programa de Destrezas para manejo ...

  17. Type III restriction endonucleases are heterotrimeric: comprising one helicase–nuclease subunit and a dimeric methyltransferase that binds only one specific DNA

    Science.gov (United States)

    Butterer, Annika; Pernstich, Christian; Smith, Rachel M.; Sobott, Frank; Szczelkun, Mark D.; Tóth, Júlia

    2014-01-01

    Fundamental aspects of the biochemistry of Type III restriction endonucleases remain unresolved despite being characterized by numerous research groups in the past decades. One such feature is the subunit stoichiometry of these hetero-oligomeric enzyme complexes, which has important implications for the reaction mechanism. In this study, we present a series of results obtained by native mass spectrometry and size exclusion chromatography with multi-angle light scattering consistent with a 1:2 ratio of Res to Mod subunits in the EcoP15I, EcoPI and PstII complexes as the main holoenzyme species and a 1:1 stoichiometry of specific DNA (sDNA) binding by EcoP15I and EcoPI. Our data are also consistent with a model where ATP hydrolysis activated by recognition site binding leads to release of the enzyme from the site, dissociation from the substrate via a free DNA end and cleavage of the DNA. These results are discussed critically in the light of the published literature, aiming to resolve controversies and discuss consequences in terms of the reaction mechanism. PMID:24510100

  18. Home automation becomes home entertainment. Emotion replaces function; Hausautomation sucht Naehe zum Home-Entertainment. Emotion verdraengt Funktion

    Energy Technology Data Exchange (ETDEWEB)

    Schmid, W.

    2006-09-15

    Home automation commonly applies to control of lighting, shading, heating, safety etc., i.e. a purely functional concept. Recently, many systems have come to integrate home entertainment as well: Bang and Olufsen and Revox meets heating, light, and ventilation. (orig.)

  19. There is no place like @home!: The value of home consultations in paediatric rehabilitation.

    Science.gov (United States)

    van Maren-Suir, I; Ketelaar, M; Brouns, B; van der Sanden, K; Verhoef, M

    2018-07-01

    Family-centred services (FCS) is widely regarded as the best practice approach in early interventions. Creating a therapeutic environment, which also stimulates collaboration between parents and service professionals, is a way to conform to the principles of FCS. The present paper describes the project entitled @home, involving the implementation of home consultations by a specialized team working with children aged 0-5 years at our rehabilitation centre in the Netherlands. The objectives of this article are to (a) describe the development and implementation of home consultations as part of regular care and (b) share the experiences of parents and service providers with home consultations. The implementation process was divided into 3 steps: (1) interviewing experts, (2) adjusting current rehabilitation trajectories, and (3) service providers offering consultations to children at home. The experiences with the home consultations were immediately incorporated in the system, making the implementation an iterative process. In 82% of the 133 home conducted consultations, the service professionals reported that it was more valuable to offer home consultations than seeing the child at the rehabilitation centre. The semistructured interviews revealed that parents and service providers found that they received and provided more tailored advice, perceived a more equal partnership between service professionals and parents, and reported that the home consultations provided a good natural therapeutic environment where a child can be itself and where the child performs best. By using the @home system based on the 3 service models, home consultations are now part of the regular paediatric rehabilitation system at our rehabilitation centre. © 2018 John Wiley & Sons Ltd.

  20. Pervasive Home Care - Technological support for treatment of diabetic foot ulcers at home

    DEFF Research Database (Denmark)

    Larsen, Simon Bo

    2006-01-01

    of the patient in collaboration with patient and home care clinicians. My main research method has been qualitative analysis of the empirical results generated during an experimental project using Participatory Design (PD) to investigate potential futures in the treatment of patients with diabetic foot ulcers...... the need arises for moving treatment and care involving specialised knowledge from the hospital to the home. In this dissertation I use the term Home Care" for the multidisciplinary investigation of how this movement can be supported with technology enabling the expert to carry on a treatment in the home...... approach that I outline in this dissertation. Furthermore I describe the results of the project contributing to three related scientific fields: home care technologies, telemedicine and computer supported cooperative work (CSCW). The main conclusion towards home care technologies is that the many visionary...

  1. Exploring new operational research opportunities within the Home Care context: the chemotherapy at home.

    Science.gov (United States)

    Chahed, Salma; Marcon, Eric; Sahin, Evren; Feillet, Dominique; Dallery, Yves

    2009-06-01

    Home Care (HC) services provide complex and coordinated medical and paramedical care to patients at their homes. As health care services move into the home setting, the need for developing innovative approaches that improve the efficiency of home care organizations increases. We first conduct a literature review of investigations dealing with operation planning within the area of home care management. We then address a particular issue dealing with the planning of operations related to chemotherapy at home as it is an emergent problem in the French context. Our interest is focused on issues specific to the anti-cancer drug supply chain. We identify various models that can be developed and analyze one of them.

  2. The Science of Home Automation

    Science.gov (United States)

    Thomas, Brian Louis

    Smart home technologies and the concept of home automation have become more popular in recent years. This popularity has been accompanied by social acceptance of passive sensors installed throughout the home. The subsequent increase in smart homes facilitates the creation of home automation strategies. We believe that home automation strategies can be generated intelligently by utilizing smart home sensors and activity learning. In this dissertation, we hypothesize that home automation can benefit from activity awareness. To test this, we develop our activity-aware smart automation system, CARL (CASAS Activity-aware Resource Learning). CARL learns the associations between activities and device usage from historical data and utilizes the activity-aware capabilities to control the devices. To help validate CARL we deploy and test three different versions of the automation system in a real-world smart environment. To provide a foundation of activity learning, we integrate existing activity recognition and activity forecasting into CARL home automation. We also explore two alternatives to using human-labeled data to train the activity learning models. The first unsupervised method is Activity Detection, and the second is a modified DBSCAN algorithm that utilizes Dynamic Time Warping (DTW) as a distance metric. We compare the performance of activity learning with human-defined labels and with automatically-discovered activity categories. To provide evidence in support of our hypothesis, we evaluate CARL automation in a smart home testbed. Our results indicate that home automation can be boosted through activity awareness. We also find that the resulting automation has a high degree of usability and comfort for the smart home resident.

  3. Prescribing quality for older people in Norwegian nursing homes and home nursing services using multidose dispensed drugs.

    Science.gov (United States)

    Halvorsen, Kjell H; Granas, Anne Gerd; Engeland, Anders; Ruths, Sabine

    2012-09-01

    To examine and compare the quality of drug prescribing for older patients in nursing homes and home nursing services. Cross-sectional study comprising 11,254 patients aged ≥ 65 years in nursing homes (n = 2986) and home nursing services (n = 8268). Potentially inappropriate medications were identified by using the Norwegian General Practice criteria and drug-drug interactions through a Norwegian Web-based tool. The impact of care setting on exposure to selected drug groups, potentially inappropriate medications, and drug interactions was calculated, adjusting for patients' age, gender, and number of drugs used. Patients in nursing homes and home nursing services used on average 5.7 (SD = 2.6) multidose dispensed regular drugs. Twenty-six percent used at least one potentially inappropriate medication, 31% in nursing homes and 25% in home nursing services, p nursing homes (18%) and home nursing services (9%), p nursing homes, more patients in home nursing services used cardiovascular drugs and fewer patients used psychotropic drugs. Altogether, 8615 drug-drug interactions were identified in 55% of patients, 48% in nursing homes and 57% in home nursing services, p quality of drug prescribing in nursing homes compared with home nursing services. Explanations as to why these differences exist need to be further explored. Copyright © 2011 John Wiley & Sons, Ltd.

  4. Tactics for homing in mobile life

    DEFF Research Database (Denmark)

    Petersen, Marianne Graves; Lynggard, Aviaja Borup; Krogh, Peter Gall

    2010-01-01

    For many people home making is an activity, which extends beyond a single house. We introduce the terminology of Homing as the act of home making, when in a primary home, secondary home or more temporary spaces. By point of departure in existing literature on home making and through ethnographic...

  5. Windows Home Server users guide

    CERN Document Server

    Edney, Andrew

    2008-01-01

    Windows Home Server brings the idea of centralized storage, backup and computer management out of the enterprise and into the home. Windows Home Server is built for people with multiple computers at home and helps to synchronize them, keep them updated, stream media between them, and back them up centrally. Built on a similar foundation as the Microsoft server operating products, it's essentially Small Business Server for the home.This book details how to install, configure, and use Windows Home Server and explains how to connect to and manage different clients such as Windows XP, Windows Vist

  6. 14 GHz longitudinally detected electron spin resonance using microHall sensors

    Science.gov (United States)

    Bouterfas, M.; Mouaziz, S.; Popovic, R. S.

    2017-09-01

    In this work we developed a home-made LOngitudinally Detected Electron Spin Resonance (LODESR) spectrometer based on a microsize Hall sensor. A coplanar waveguide (CPW)-resonator is used to induce microwave-excitation on the sample at 14 GHz. We used InSb cross-shaped Hall devices with active areas of (10 μm × 10 μm) and (5 μm × 5 μm) . Signal intensities of the longitudinal magnetization component of DPPH and YIG samples of volumes about (10 μm) 3 and (5 μm) 3 , are measured under amplitude and frequency modulated microwave magnetic field generated by the CPW-resonator. At room temperature, 109spins /G √Hz sensitivity is achieved for 0.2mT linewidth, a result which is still better than most of inductive detected LODESR sensitivities.

  7. Comparison of Long-term Care in Nursing Homes Versus Home Health: Costs and Outcomes in Alabama.

    Science.gov (United States)

    Blackburn, Justin; Locher, Julie L; Kilgore, Meredith L

    2016-04-01

    To compare acute care outcomes and costs among nursing home residents with community-dwelling home health recipients. A matched retrospective cohort study of Alabamians aged more than or equal to 65 years admitted to a nursing home or home health between March 31, 2007 and December 31, 2008 (N = 1,291 pairs). Medicare claims were compared up to one year after admission into either setting. Death, emergency department and inpatient visits, inpatient length of stay, and acute care costs were compared using t tests. Medicaid long-term care costs were compared for a subset of matched beneficiaries. After one year, 77.7% of home health beneficiaries were alive compared with 76.2% of nursing home beneficiaries (p Home health beneficiaries averaged 0.2 hospital visits and 0.1 emergency department visits more than nursing home beneficiaries, differences that were statistically significant. Overall acute care costs were not statistically different; home health beneficiaries' costs averaged $31,423, nursing home beneficiaries' $32,239 (p = .5032). Among 426 dual-eligible pairs, Medicaid long-term care costs averaged $4,582 greater for nursing home residents (p nursing home or home health care. Additional research controlling for exogenous factors relating to long-term care decisions is needed. © The Author 2014. Published by Oxford University Press on behalf of The Gerontological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  8. Smart Home Test Bed: Examining How Smart Homes Interact with the Power Grid

    Energy Technology Data Exchange (ETDEWEB)

    2016-11-01

    This fact sheet highlights the Smart Home Test Bed capability at the Energy Systems Integration Facility. The National Renewable Energy Laboratory (NREL) is working on one of the new frontiers of smart home research: finding ways for smart home technologies and systems to enhance grid operations in the presence of distributed, clean energy technologies such as photovoltaics (PV). To help advance this research, NREL has developed a controllable, flexible, and fully integrated Smart Home Test Bed.

  9. Home in the Making

    DEFF Research Database (Denmark)

    Kreuzer, Maria; von Wallpach, Sylvia; Muehlbacher, Hans

    2016-01-01

    In a context of unprecedented migration home reaches high relevance. This study aims at understanding the (re-)construction of home by first generation consumer migrants. The findings provide insights into consumers’ (re-)construction of various dimensions of home and identify “inner home” as a n...

  10. Smart assistants for smart homes

    OpenAIRE

    Rasch, Katharina

    2013-01-01

    The smarter homes of tomorrow promise to increase comfort, aid elderly and disabled people, and help inhabitants save energy. Unfortunately, smart homes today are far from this vision – people who already live in such a home struggle with complicated user interfaces, inflexible home configurations, and difficult installation procedures. Under these circumstances, smart homes are not ready for mass adoption. This dissertation addresses these issues by proposing two smart assistants for smart h...

  11. DOE Zero Energy Ready Home Case Study: TC Legend Homes, Seattle, Washington

    Energy Technology Data Exchange (ETDEWEB)

    none,

    2013-09-01

    This house incorporates slab-on-grade, EPS roof, and radiant heating with an air-to-water heat pump that also preheats domestic hot water. Without counting in the solar panels, the home earns a home energy rating system (HERS) score of 37, with projected utility bills of about $740 a year. With the 6.4-kW photovoltaic power system installed on the roof, the home’s HERS scores drops to -1 and utility bills for the all-electric home drop to zero. This home was awarded a 2013 Housing Innovation Award in the affordable builder category.

  12. Research on the model of home networking

    Science.gov (United States)

    Yun, Xiang; Feng, Xiancheng

    2007-11-01

    It is the research hotspot of current broadband network to combine voice service, data service and broadband audio-video service by IP protocol to transport various real time and mutual services to terminal users (home). Home Networking is a new kind of network and application technology which can provide various services. Home networking is called as Digital Home Network. It means that PC, home entertainment equipment, home appliances, Home wirings, security, illumination system were communicated with each other by some composing network technology, constitute a networking internal home, and connect with WAN by home gateway. It is a new network technology and application technology, and can provide many kinds of services inside home or between homes. Currently, home networking can be divided into three kinds: Information equipment, Home appliances, Communication equipment. Equipment inside home networking can exchange information with outer networking by home gateway, this information communication is bidirectional, user can get information and service which provided by public networking by using home networking internal equipment through home gateway connecting public network, meantime, also can get information and resource to control the internal equipment which provided by home networking internal equipment. Based on the general network model of home networking, there are four functional entities inside home networking: HA, HB, HC, and HD. (1) HA (Home Access) - home networking connects function entity; (2) HB (Home Bridge) Home networking bridge connects function entity; (3) HC (Home Client) - Home networking client function entity; (4) HD (Home Device) - decoder function entity. There are many physical ways to implement four function entities. Based on theses four functional entities, there are reference model of physical layer, reference model of link layer, reference model of IP layer and application reference model of high layer. In the future home network

  13. Home Health Compare

    Data.gov (United States)

    U.S. Department of Health & Human Services — Home Health Compare has information about the quality of care provided by Medicare-certified home health agencies throughout the nation. Medicare-certified means the...

  14. PLANNED HOME BIRTH: A REVIEW

    Directory of Open Access Journals (Sweden)

    Tamara Serdinšek

    2016-05-01

    Full Text Available Background: Home birth is as old as humanity, but still most middle- and high-income countries consider hospitals as the safest birth settings, as complications regarding birth are highly unpredictable. Despite this there are a few countries in which home birth in integrated into official healthcare system (the Netherlands, United Kingdom, Canada etc.. Home births can be divided into unplanned and planned, and the latter can be further categorized by the presence of the birth attendants. This review focuses on planned home births, which are differently represented throughout the world. In the United States 0.6-1.0% of all children are born at home, in the United Kingdom 2-3%, in Canada 1.6% and in the Netherlands 20-30%. For Slovenia, the number of planned home births is unknown; however, in 2010 0.1% of children were born outside medical facilities.Conclusions: The safety of home birth in still under the debate. While research confirms smaller number of obstetric interventions and some complications in mothers who give birth at home, the data regarding the neonatal and perinatal mortality and morbidity is still conflicting. This confirms the need for large multicentric trials in this field. Current home birth guidelines emphasize that women should be well informed regarding the possible advantages and disadvantages of home births. In addition, the emphasis is on definition of selection criteria for home birth, indications for intrapartal transfer to the hospital and appropriate education of birth attendants. 

  15. DOE Zero Energy Ready Home Case Study: Amaris Custom Homes, St. Paul, Minnesota

    Energy Technology Data Exchange (ETDEWEB)

    None

    2015-06-01

    For this project, Amaris worked with U.S. Department of Energy (DOE) team, NorthernSTAR Building America Partnership, to develop the first Zero Energy Ready Home (ZERH) in Minnesota's cold climate using reasonable, cost-effective, and replicable construction materials and practices. The result is a passive solar, super-efficient 3542-ft2 walkout ranch-style home with all the creature comforts. Along with meeting ZERH standards, Amaris also achieved certifications for Leadership in Energy & Environmental Design for Homes v4, MN Green Path Emerald, and a Builders Association of the Twin Cities Reggie Award of Excellence. The home achieves a HERS score of 41 without photovoltaics; with PV, the home achieves a HERS score of 5.

  16. Home financing for new and existing energy efficient homes

    International Nuclear Information System (INIS)

    1994-01-01

    The video describes how the home federal savings and loan association in Rockford, IL, has developed lending standards for new conventional and solar homes. The nine standards are illustrated. The video follows the customer through the loan process, including an appraisal of energy efficient items, loan closing, and continuing education. The primary audience is savings and loan management personnel

  17. On the Design of Smart Homes: A Framework for Activity Recognition in Home Environment.

    Science.gov (United States)

    Cicirelli, Franco; Fortino, Giancarlo; Giordano, Andrea; Guerrieri, Antonio; Spezzano, Giandomenico; Vinci, Andrea

    2016-09-01

    A smart home is a home environment enriched with sensing, actuation, communication and computation capabilities which permits to adapt it to inhabitants preferences and requirements. Establishing a proper strategy of actuation on the home environment can require complex computational tasks on the sensed data. This is the case of activity recognition, which consists in retrieving high-level knowledge about what occurs in the home environment and about the behaviour of the inhabitants. The inherent complexity of this application domain asks for tools able to properly support the design and implementation phases. This paper proposes a framework for the design and implementation of smart home applications focused on activity recognition in home environments. The framework mainly relies on the Cloud-assisted Agent-based Smart home Environment (CASE) architecture offering basic abstraction entities which easily allow to design and implement Smart Home applications. CASE is a three layered architecture which exploits the distributed multi-agent paradigm and the cloud technology for offering analytics services. Details about how to implement activity recognition onto the CASE architecture are supplied focusing on the low-level technological issues as well as the algorithms and the methodologies useful for the activity recognition. The effectiveness of the framework is shown through a case study consisting of a daily activity recognition of a person in a home environment.

  18. Prediction of Advisability of Returning Home Using the Home Care Score

    Directory of Open Access Journals (Sweden)

    Akiyoshi Matsugi

    2015-01-01

    Full Text Available Purpose. The aim of this study was to assess whether the home care score (HCS, which was developed by the Ministry of Health and Welfare in Japan in 1992, is useful for the prediction of advisability of home care. Methods. Subjects living at home and in assisted-living facilities were analyzed. Binominal logistic regression analyses, using age, sex, the functional independence measure score, and the HCS, along with receiver operating characteristic curve analyses, were conducted. Findings/Conclusions. Only HCS was selected for the regression equation. Receiver operating characteristic curve analysis revealed that the area under the curve (0.9, sensitivity (0.82, specificity (0.83, and positive predictive value (0.84 for HCS were higher than those for the functional independence measure, indicating that the HCS is a powerful predictor for advisability of home care. Clinical Relevance. Comprehensive measurements of the condition of provided care and the activities of daily living of the subjects, which are included in the HCS, are required for the prediction of advisability of home care.

  19. Prediction of Advisability of Returning Home Using the Home Care Score

    Science.gov (United States)

    Matsugi, Akiyoshi; Tani, Keisuke; Tamaru, Yoshiki; Yoshioka, Nami; Yamashita, Akira; Mori, Nobuhiko; Oku, Kosuke; Ikeda, Masashi; Nagano, Kiyoshi

    2015-01-01

    Purpose. The aim of this study was to assess whether the home care score (HCS), which was developed by the Ministry of Health and Welfare in Japan in 1992, is useful for the prediction of advisability of home care. Methods. Subjects living at home and in assisted-living facilities were analyzed. Binominal logistic regression analyses, using age, sex, the functional independence measure score, and the HCS, along with receiver operating characteristic curve analyses, were conducted. Findings/Conclusions. Only HCS was selected for the regression equation. Receiver operating characteristic curve analysis revealed that the area under the curve (0.9), sensitivity (0.82), specificity (0.83), and positive predictive value (0.84) for HCS were higher than those for the functional independence measure, indicating that the HCS is a powerful predictor for advisability of home care. Clinical Relevance. Comprehensive measurements of the condition of provided care and the activities of daily living of the subjects, which are included in the HCS, are required for the prediction of advisability of home care. PMID:26491568

  20. The Impact of Certificate-of-Need Laws on Nursing Home and Home Health Care Expenditures.

    Science.gov (United States)

    Rahman, Momotazur; Galarraga, Omar; Zinn, Jacqueline S; Grabowski, David C; Mor, Vincent

    2016-02-01

    Over the past two decades, nursing homes and home health care agencies have been influenced by several Medicare and Medicaid policy changes including the adoption of prospective payment for Medicare-paid postacute care and Medicaid-paid long-term home and community-based care reforms. This article examines how spending growth in these sectors was affected by state certificate-of-need (CON) laws, which were designed to limit the growth of providers and have remained unchanged for several decades. Compared with states without CON laws, Medicare and Medicaid spending in states with CON laws grew faster for nursing home care and more slowly for home health care. In particular, we observed the slowest growth in community-based care in states with CON for both the nursing home and home health industries. Thus, controlling for other factors, public postacute and long-term care expenditures in CON states have become dominated by nursing homes. © The Author(s) 2015.

  1. Home-based Healthcare Technology

    DEFF Research Database (Denmark)

    Verdezoto, Nervo

    of these systems target a specific treatment or condition and might not be sufficient to support the care management work at home. Based on a case study approach, my research investigates home-based healthcare practices and how they can inform future design of home-based healthcare technology that better account......Sustaining daily, unsupervised healthcare activities in non-clinical settings such as the private home can challenge, among others, older adults. To support such unsupervised care activities, an increasingly number of reminders and monitoring systems are being designed. However, most...

  2. Population Health and Tailored Medical Care in the Home: the Roles of Home-Based Primary Care and Home-Based Palliative Care.

    Science.gov (United States)

    Ritchie, Christine S; Leff, Bruce

    2018-03-01

    With the growth of value-based care, payers and health systems have begun to appreciate the need to provide enhanced services to homebound adults. Recent studies have shown that home-based medical services for this high-cost, high-need population reduce costs and improve outcomes. Home-based medical care services have two flavors that are related to historical context and specialty background-home-based primary care (HBPC) and home-based palliative care (HBPalC). Although the type of services provided by HBPC and HBPalC (together termed "home-based medical care") overlap, HBPC tends to encompass longitudinal and preventive care, while HBPalC often provides services for shorter durations focused more on distress management and goals of care clarification. Given workforce constraints and growing demand, both HBPC and HBPalC will benefit from working together within a population health framework-where HBPC provides care to all patients who have trouble accessing traditional office practices and where HBPalC offers adjunctive care to patients with high symptom burden and those who need assistance with goals clarification. Policy changes that support provision of medical care in the home, population health strategies that tailor home-based medical care to the specific needs of the patients and their caregivers, and educational initiatives to assure basic palliative care competence for all home-based medical providers will improve access and reduce illness burden to this important and underrecognized population. Copyright © 2017 American Academy of Hospice and Palliative Medicine. Published by Elsevier Inc. All rights reserved.

  3. Rigorous numerical study of strong microwave photon-magnon coupling in all-dielectric magnetic multilayers

    Energy Technology Data Exchange (ETDEWEB)

    Maksymov, Ivan S., E-mail: ivan.maksymov@uwa.edu.au [School of Physics M013, The University of Western Australia, 35 Stirling Highway, Crawley, WA 6009 (Australia); ARC Centre of Excellence for Nanoscale BioPhotonics, School of Applied Sciences, RMIT University, Melbourne, VIC 3001 (Australia); Hutomo, Jessica; Nam, Donghee; Kostylev, Mikhail [School of Physics M013, The University of Western Australia, 35 Stirling Highway, Crawley, WA 6009 (Australia)

    2015-05-21

    We demonstrate theoretically a ∼350-fold local enhancement of the intensity of the in-plane microwave magnetic field in multilayered structures made from a magneto-insulating yttrium iron garnet (YIG) layer sandwiched between two non-magnetic layers with a high dielectric constant matching that of YIG. The enhancement is predicted for the excitation regime when the microwave magnetic field is induced inside the multilayer by the transducer of a stripline Broadband Ferromagnetic Resonance (BFMR) setup. By means of a rigorous numerical solution of the Landau-Lifshitz-Gilbert equation consistently with the Maxwell's equations, we investigate the magnetisation dynamics in the multilayer. We reveal a strong photon-magnon coupling, which manifests itself as anti-crossing of the ferromagnetic resonance magnon mode supported by the YIG layer and the electromagnetic resonance mode supported by the whole multilayered structure. The frequency of the magnon mode depends on the external static magnetic field, which in our case is applied tangentially to the multilayer in the direction perpendicular to the microwave magnetic field induced by the stripline of the BFMR setup. The frequency of the electromagnetic mode is independent of the static magnetic field. Consequently, the predicted photon-magnon coupling is sensitive to the applied magnetic field and thus can be used in magnetically tuneable metamaterials based on simultaneously negative permittivity and permeability achievable thanks to the YIG layer. We also suggest that the predicted photon-magnon coupling may find applications in microwave quantum information systems.

  4. Improving Elderly's Dental Hygiene Through Nursing Home Staff's Dental Health Education at the Nursing Home

    OpenAIRE

    Santoso, Bedjo; Eko Ningtyas, Endah Aryati; Fatmasari, Diyah

    2017-01-01

    Stomatitis often occurs in elderly at nursing home. They need nursing home staff assistance to maintain their dental and oral health. Therefore, nursing home staff need dental health education. Lecture or discussion methods, which are more effective to improve knowledge, attitude and skill of nursing home staff was the purpose of this research. The research design was quasi-experiment research and pretest-posttest with control group. The sample was 42 nursing home staffs and 74 elderlies, div...

  5. Home Within Me

    DEFF Research Database (Denmark)

    Kreuzer, Maria; Mühlbacher, Hans; von Wallpach, Sylvia

    2017-01-01

    In an increasingly globalized, digitalized and perceived unmanageable world, consumers strive for belongingness, identification and security and re-discover the importance of home. Home is central to peoples’ individual as well as collective identities and their self-development (McCracken, 1989...... in Austria and the sample consisted of 15 locals (study 1) and 17 first generation immigrants (study 2) to identify possible commonalities and differences. This research adds to existing literature by 1) empirically confirming the existence of dimensions of home (e.g., physical, social, temporary...

  6. HOME Rent Limits

    Data.gov (United States)

    Department of Housing and Urban Development — In accordance with 24 CFR Part 92.252, HUD provides maximum HOME rent limits. The maximum HOME rents are the lesser of: The fair market rent for existing housing for...

  7. Home and Office Blood Pressure Control among Treated Hypertensive Patients in Japan: Findings from the Japan Home versus Office Blood Pressure Measurement Evaluation (J-HOME Study

    Directory of Open Access Journals (Sweden)

    Nariyasu Mano

    2010-02-01

    Full Text Available Appropriate control of blood pressure (BP is essential for prevention of future cardiovascular events. However, BP control among treated hypertensive patients has been insufficient. Recently, the usefulness of self-measured BP at home (home BP measurement for the management of hypertension has been reported in many studies. We evaluated BP control both at home and in the office among treated hypertensive patients in primary care settings in Japan (the J-HOME study. We found poor control of home and office BPs and clarified some factors affecting control. We also examined factors associated with the magnitude of the white-coat effect, the morning–evening BP difference, and home heart rate in this J-HOME study.

  8. Pervasive Home Environments

    Science.gov (United States)

    Bull, P.; Limb, R.; Payne, R.

    An increasing number of computers and other equipment, such as games consoles and multimedia appliances for the home, have networking capability. The rapid growth of broadband in the home is also fuelling the demand for people to network their homes. In the near future we will see a number of market sectors trying to 'own' the home by providing gateways either from the traditional ISP or from games and other service providers. The consumer is bombarded with attractive advertising to acquire the latest technological advances, but is left with a plethora of different appliances, which have a bewildering range of requirements and features in terms of networking, user interface, and higher-level communications protocols. In many cases, these are proprietary, preventing interworking. Such technical and usability anarchy confuses the consumer and could ultimately suppress market adoption.

  9. Vascular access for home haemodialysis.

    Science.gov (United States)

    Al Shakarchi, Julien; Day, C; Inston, N

    2018-03-01

    Home haemodialysis has been advocated due to improved quality of life. However, there are very little data on the optimum vascular access for it. A retrospective cohort study was carried on all patients who initiated home haemodialysis between 2011 and 2016 at a large university hospital. Access-related hospital admissions and interventions were used as primary outcome measures. Our cohort consisted of 74 patients. On initiation of home haemodialysis, 62 individuals were using an arteriovenous fistula as vascular access, while the remaining were on a tunnelled dialysis catheter. Of the 12 patients who started on a tunnelled dialysis catheter, 5 were subsequently converted to either an arteriovenous fistula ( n = 4) or an arteriovenous graft ( n = 1). During the period of home haemodialysis use, four arteriovenous fistula failed or thrombosed with patients continuing on home haemodialysis using an arteriovenous graft ( n = 3) or a tunnelled dialysis catheter ( n = 1). To maintain uninterrupted home haemodialysis, interventional rates were 0.32 per arteriovenous fistula/arteriovenous graft access-year and 0.4 per tunnelled dialysis catheter access-year. Hospital admission rates for patients on home haemodialysis were 0.33 per patient-year. Our study has shown that home haemodialysis can be safely and independently performed at home within a closely managed home haemodialysis programme. The authors also advocate the use of arteriovenous fistulas for this cohort of patients due to both low complication and intervention rates.

  10. DOE Zero Energy Ready Home Case Study: Mandalay Homes, Phoenix, Arizona

    Energy Technology Data Exchange (ETDEWEB)

    none,

    2013-09-01

    This builder built fourteen homes in the Gordon Estates subdivision that achieved Challenge Home certification with HERS 38–58 on an affordable budget for homeowners. Every Mandalay home in the development also met the National Green Building Standard gold level. The Gordon Estates subdivision is also serving as a showcase of energy efficiency, and Mandalay is hosting education workshops for realtors, state and local officials, other builders, students, potential homeowners, and the public. The builder won a 2013 Housing Innovation Award in the affordable builder category.

  11. Smart homes with smartphones : Creating a Smart home application for smartphones

    OpenAIRE

    Lärka, Martin

    2015-01-01

    The goal of this master thesis was to evaluate the field of Smart home applications run on mobile platforms such as smart phones. This was at the time a relatively new field that has attracted the attention of Smart phone giants like Apple and Google. This raised the question whether or not the users willingness to embrace the technology. To evaluate the field, a literature study was conducted covering the Smart home technology, and Apple’s and Google’s Smart home solutions. The rendered in a...

  12. DOE Zero Energy Ready Home Case Study, Weiss Building & Development, LLC., System Home, River Forest, Illinois

    Energy Technology Data Exchange (ETDEWEB)

    none,

    2013-09-01

    The Passive House Challenge Home located in River Forest, Illinois, is a 5-bedroom, 4.5-bath, 3,600 ft2 two-story home (plus basement) that costs about $237 less per month to operate than a similar sized home built to the 2009 IECC. For a home with no solar photovoltaic panels installed, it scored an amazingly low 27 on the Home Energy Rating System (HERS) score.An ENERGY STAR-rated dishwasher, clothes washer, and refrigerator; an induction cooktop, condensing clothes dryer, and LED lighting are among the energy-saving devices inside the home. All plumbing fixtures comply with EPA WaterSense criteria. The home was awarded a 2013 Housing Innovation Award in the "systems builder" category.

  13. Genetics Home Reference

    Science.gov (United States)

    ... Page Search Home Health Conditions Genes Chromosomes & mtDNA Resources Help Me Understand Genetics Share: Email Facebook Twitter Genetics Home Reference provides consumer-friendly information about the effects of genetic variation on human health. Health Conditions More than 1,200 health ...

  14. LHC@home gets new home

    CERN Multimedia

    Oates, John

    2007-01-01

    "The distributed computing project LHC@home is moving to London from Cern in Switzerland. Researchers at Qeen Mary University have been trialling the system since June, but are now ready for the offical launch" (1 page)

  15. Home Appliances as Home Controllers: Concepts and Set-Top Box Implementation

    Directory of Open Access Journals (Sweden)

    Z. Jovanovic

    2010-11-01

    Full Text Available In this paper we present a novel softwarebased home control platform suitable as an extension to digital home appliances that are equipped with a CPU (settop boxes, home theatre systems, TV sets, gaming consoles, etc. By using an appliance they are already accustomed to, users become able to control lights, appliances and media playback in their homes. Intelligence and awareness are achieved with a support for execution of recipes – preprepared scripts that define timely actions and respond to triggers obtained from sensors. Software abstraction layer facilitates integration of any desired communication protocol. In our prototype, we supported Zigbee and DMX for light control, X10 for light/appliances control over power line, as well as Ethernet-based optical cameras as motion/presence sensors and UPnP/DLNA based equipment for distributed media playback.

  16. Environmental impacts of informal settlements with second homes (vacation homes. A case study: Apuseni Nature Park.

    Directory of Open Access Journals (Sweden)

    OANA ALEXANDRA CIUPE

    2017-10-01

    Full Text Available The past decades has seen the rapid development of second homes in many country of the world. Therefore, secondary dwellings used for tourism-related purposes (vacation homes or second homes are a reality that becomes more pronounced and visible at both nationally and internationally level. However, the rapidity and the novelty of the phenomenon creates difficulties in terms of efficient management and suitable integration in spatial planning and urbanism plans, favoring - in a negative way - expanding of build-up areas (with second homes in a chaotic way. Since there has been no detailed investigation of second homes tourism from the perspective of informal settlements, this article follows a case-study design, with in-depth analysis of informal settlements with vacation homes found in Apuseni Nature Park. Based on long-term field research, will be exemplified 4 types of informal settlements with second homes (vacation homes identified in the case study. The aim of this paper is to critically analyse the effects of informal settlements with vacation homes on the natural environment.

  17. Comparison of home and away-from-home physical activity using accelerometers and cellular network-based tracking devices.

    Science.gov (United States)

    Ramulu, Pradeep Y; Chan, Emilie S; Loyd, Tara L; Ferrucci, Luigi; Friedman, David S

    2012-08-01

    Measuring physical at home and away from home is essential for assessing health and well-being, and could help design interventions to increase physical activity. Here, we describe how physical activity at home and away from home can be quantified by combining information from cellular network-based tracking devices and accelerometers. Thirty-five working adults wore a cellular network-based tracking device and an accelerometer for 6 consecutive days and logged their travel away from home. Performance of the tracking device was determined using the travel log for reference. Tracking device and accelerometer data were merged to compare physical activity at home and away from home. The tracking device detected 98.6% of all away-from-home excursions, accurately measured time away from home and demonstrated few prolonged signal drop-out periods. Most physical activity took place away from home on weekdays, but not on weekends. Subjects were more physically active per unit of time while away from home, particularly on weekends. Cellular network-based tracking devices represent an alternative to global positioning systems for tracking location, and provide information easily integrated with accelerometers to determine where physical activity takes place. Promoting greater time spent away from home may increase physical activity.

  18. Identification of human flap endonuclease 1 (FEN1) inhibitors using a machine learning based consensus virtual screening.

    Science.gov (United States)

    Deshmukh, Amit Laxmikant; Chandra, Sharat; Singh, Deependra Kumar; Siddiqi, Mohammad Imran; Banerjee, Dibyendu

    2017-07-25

    Human Flap endonuclease1 (FEN1) is an enzyme that is indispensable for DNA replication and repair processes and inhibition of its Flap cleavage activity results in increased cellular sensitivity to DNA damaging agents (cisplatin, temozolomide, MMS, etc.), with the potential to improve cancer prognosis. Reports of the high expression levels of FEN1 in several cancer cells support the idea that FEN1 inhibitors may target cancer cells with minimum side effects to normal cells. In this study, we used large publicly available, high-throughput screening data of small molecule compounds targeted against FEN1. Two machine learning algorithms, Support Vector Machine (SVM) and Random Forest (RF), were utilized to generate four classification models from huge PubChem bioassay data containing probable FEN1 inhibitors and non-inhibitors. We also investigated the influence of randomly selected Zinc-database compounds as negative data on the outcome of classification modelling. The results show that the SVM model with inactive compounds was superior to RF with Matthews's correlation coefficient (MCC) of 0.67 for the test set. A Maybridge database containing approximately 53 000 compounds was screened and top ranking 5 compounds were selected for enzyme and cell-based in vitro screening. The compound JFD00950 was identified as a novel FEN1 inhibitor with in vitro inhibition of flap cleavage activity as well as cytotoxic activity against a colon cancer cell line, DLD-1.

  19. Formal home help services and institutionalization

    DEFF Research Database (Denmark)

    Yamada, Yukari; Siersma, Volkert; Avlund, Kirsten

    2012-01-01

    The effect of home help services has been inconsistent. Raising the hypothesis that receiving small amounts of home help may postpone or prevent institutionalization, the aim of the present study is to analyze how light and heavy use of home help services was related to the risk...... for institutionalization. The study was a secondary analysis of a Danish intervention study on preventive home visits in 34 municipalities from 1999 to 2003, including 2642 home-dwelling older people who were nondisabled and did not receive public home help services at baseline in 1999 and who lived at home 18 months...... after baseline. Cox regression analysis showed that those who received home help services during the first 18 months after baseline were at higher risk of being institutionalized during the subsequent three years than those who did not receive such services. However, receiving home help for less than 1h...

  20. European Home Energy

    DEFF Research Database (Denmark)

    Tommerup, Henrik M.

    2009-01-01

    An important aim of the european energy performance of buildings directive is to improve the overall energy efficiency of new homes......An important aim of the european energy performance of buildings directive is to improve the overall energy efficiency of new homes...

  1. Mobile home automation-merging mobile value added services and home automation technologies

    OpenAIRE

    Rosendahl, Andreas; Hampe, Felix J.; Botterweck, Goetz

    2007-01-01

    non-peer-reviewed In this paper we study mobile home automation, a field that emerges from an integration of mobile application platforms and home automation technologies. In a conceptual introduction we first illustrate the need for such applications by introducing a two-dimensional conceptual model of mobility. Subsequently we suggest an architecture and discuss different options of how a user might access a mobile home automation service and the controlled devices. As another contrib...

  2. Two models of distribution of sites sensitive to the endonuclease from Micrococcus luteus in the DNA of UV irradiated Escherichia coli B/r Hcr-

    International Nuclear Information System (INIS)

    Kleibl, K.; Sedliakova, M.

    1984-01-01

    Cells prelabelled with 14 C-thymine and irradiated with 5 J/m 2 were at various intervals after UV labelled with 3 H-thymidine then treated with the extract from M. luteus and DNA was analyzed in alkaline sucrose gradients. Loss of endonuclease sensitive sites (Es sites) from the parental DNA and their occurrence in the daughter DNA were followed for at least three replication cycles. Data obtained indicate that about 50% of Es sites were lost during the first replication cycle but no additional loss was observed during subsequent cycles. Thus our data do not support a hypothesis that a half of the dimers are transferred from the parental into the daughter strands at each replication cycle. They rather indicate that dimers remain in situ and distortions accompanying dimers are distinguished either on the side of the parental or on the side of the daughter strands with an equal probability. (author)

  3. The work-home interface : The role of home-based predictors of burnout among mothers

    NARCIS (Netherlands)

    Missler, M.A.; Stroebe, M.S.; van der Laan, G.

    2014-01-01

    Research into the work–home interface has mainly focused on work-related variables, leaving aspects associated with the home domain relatively understudied. This investigation examined both facilitation and conflict between home and work in a sample of 260 working mothers with children up to four

  4. Exploring workplace violence among home care workers in a consumer-driven home health care program.

    Science.gov (United States)

    Nakaishi, Lindsay; Moss, Helen; Weinstein, Marc; Perrin, Nancy; Rose, Linda; Anger, W Kent; Hanson, Ginger C; Christian, Mervyn; Glass, Nancy

    2013-10-01

    Nominal research has examined sexual harassment and workplace violence against home care workers within consumer-driven home care models such as those offered in Oregon. This study examined home care workers' experiences of violence while providing care to consumer employers, the patients who hire and manage home care workers. Focus groups and interviews were conducted in Oregon with 83 home care workers, 99 Oregon Department of Human Services (DHS) employees, and 11 consumer employers. Home care workers reported incidents of workplace physical violence (44%), psychological abuse (65%), sexual harassment (41%), and sexual violence (14%). Further, three themes were identified that may increase the risk of workplace violence: (1) real and perceived barriers to reporting violence; (2) tolerance of violence; and (3) limited training to prevent violence. To ensure worker safety while maintaining quality care, safety policies and training for consumer employers, state DHS employees, and home care workers must be developed. Copyright 2013, SLACK Incorporated.

  5. Involvement in decisions about intravenous treatment for nursing home patients: nursing homes versus hospital wards.

    Science.gov (United States)

    Klomstad, Kristin; Pedersen, Reidar; Førde, Reidun; Romøren, Maria

    2018-05-08

    Many of the elderly in nursing homes are very ill and have a reduced quality of life. Life expectancy is often hard to predict. Decisions about life-prolonging treatment should be based on a professional assessment of the patient's best interest, assessment of capacity to consent, and on the patient's own wishes. The purpose of this study was to investigate and compare how these types of decisions were made in nursing homes and in hospital wards. Using a questionnaire, we studied the decision-making process for 299 nursing home patients who were treated for dehydration using intravenous fluids, or for bacterial infections using intravenous antibiotics. We compared the 215 (72%) patients treated in nursing homes to the 84 (28%) nursing home patients treated in the hospital. The patients' capacity to consent was considered prior to treatment in 197 (92%) of the patients treated in nursing homes and 56 (67%) of the patients treated in hospitals (p nursing homes than in hospital (90% vs. 52%). Next of kin and other health personnel were also more rarely involved when the nursing home patient was treated in hospital. Whether advance care planning had been carried out, was more often unknown in the hospital (69% vs. 17% in nursing homes). Hospital doctors expressed more doubt about the decision to admit the patient to the hospital than about the treatment itself. This study indicates a potential for improvement in decision-making processes in general, and in particular when nursing home patients are treated in a hospital ward. The findings corroborate that nursing home patients should be treated locally if adequate health care and treatment is available. The communication between the different levels of health care when hospitalization is necessary, must be better. ClinicalTrials.gov NCT01023763 (12/1/09) [The registration was delayed one month after study onset due to practical reasons].

  6. FastStats: Home Health Care

    Science.gov (United States)

    ... this? Submit What's this? Submit Button NCHS Home Home Health Care Recommend on Facebook Tweet Share Compartir Data are ... National Study of Long-Term Care Providers Nursing Home Care Residential Care Communities Centers for Medicare and Medicaid ...

  7. Comparing Sleep Quality and General Health Among the Elderly Living at Home and at Nursing Home

    Directory of Open Access Journals (Sweden)

    Mansour Beyrami

    2014-01-01

    Full Text Available Objectives: Survey about the issues and problems related to elderly in order to improve their quality of life of this increasing population has become a universal concern.This study was performed by the purpose of comparing the sleep quality and general health among the Elderly Residing at Home and Old People's Homes. Methods & Materials: This study is descriptive-analytic type. Population of this investigation consisted of elderly men and women (upper than 60 years old living at personal home and at nursing home in Tabriz. Sample group composed of 100 elderly (50 men and 50 women 50 living at home and 50 living at nursing home who were selected through available sampling method. For collecting data, Goldberg General Health Questionnaire and Pittsburgh Sleep Quality Index were used. Data were analyzed by Multivariate analysis of variance (MANOVA. Results: Findings showed that In terms of general health and its components (Physical symptoms, anxiety, social dysfunction and depression and Sleep quality and its components (Subjective quality of sleep, time for sleep, total sleep time, sleep efficiency, routine, sleep disorders, sleep medications and daily dysfunction there were significant differences between nursing home residents and elderly residents in nursing homes (P=0.001. Conclusion: Findings indicated that elderly residents in nursing home are experiencing more symptoms of anxiety, depression, physical symptoms and social dysfunction Compared with the elderly whom resident at home. Also the results showed that the elderly residents of nursing homes have poor sleep quality than ones whom residents at home. On the other hand Future development of elderly care institution is inevitable. Therefore, more attention to the living conditions of elderly residents of institutions seems necessary.

  8. Foraging optimally for home ranges

    Science.gov (United States)

    Mitchell, Michael S.; Powell, Roger A.

    2012-01-01

    Economic models predict behavior of animals based on the presumption that natural selection has shaped behaviors important to an animal's fitness to maximize benefits over costs. Economic analyses have shown that territories of animals are structured by trade-offs between benefits gained from resources and costs of defending them. Intuitively, home ranges should be similarly structured, but trade-offs are difficult to assess because there are no costs of defense, thus economic models of home-range behavior are rare. We present economic models that predict how home ranges can be efficient with respect to spatially distributed resources, discounted for travel costs, under 2 strategies of optimization, resource maximization and area minimization. We show how constraints such as competitors can influence structure of homes ranges through resource depression, ultimately structuring density of animals within a population and their distribution on a landscape. We present simulations based on these models to show how they can be generally predictive of home-range behavior and the mechanisms that structure the spatial distribution of animals. We also show how contiguous home ranges estimated statistically from location data can be misleading for animals that optimize home ranges on landscapes with patchily distributed resources. We conclude with a summary of how we applied our models to nonterritorial black bears (Ursus americanus) living in the mountains of North Carolina, where we found their home ranges were best predicted by an area-minimization strategy constrained by intraspecific competition within a social hierarchy. Economic models can provide strong inference about home-range behavior and the resources that structure home ranges by offering falsifiable, a priori hypotheses that can be tested with field observations.

  9. FastStats: Nursing Home Care

    Science.gov (United States)

    ... Submit What's this? Submit Button NCHS Home Nursing Home Care Recommend on Facebook Tweet Share Compartir Data are ... Person’s Health Related Links Adult Day Services Centers Home Health Care Hospice Care National Study of Long-Term Care ...

  10. HOME CARE NURSES’ ROLES IN ENHANCING QUALITY OF NURSING CARE FOR PATIENTS AT HOME: A PHENOMENOLOGICAL STUDY

    Directory of Open Access Journals (Sweden)

    Titan Ligita

    2017-08-01

    Full Text Available Background: Provision of health care service at home is one of the advanced forms of care for patients being discharged from hospitalization. Little is known about the experience of nurses providing home care services through a nursing home-care model especially in Indonesian context. Objective: This study aims to explore the experience in order to increase understanding on the form of home care provision, and consequently the nurses may understand the form of home care globally. Methods: This study employed a phenomenological design and performed interview in the process of data collection. Data were analysed by using content analysis. Results: The main contexts of home care nurse experiences were generated. There were definition and role of home care nurses, the involvement of family members in the provision of care, the facilitating and hindering factors contributed to home care provision as well as manual on providing home care nursing. Conclusion: The implication from this study is that nursing care should be given to the patients continuously and consequently the need for family involvement is important. Additionally, in providing the home care, a proper manual is needed by home care nurses as the guidance to give best quality of care to patients.

  11. A Study of the Association Between Multidisciplinary Home Care and Home Death Among Thai Palliative Care Patients.

    Science.gov (United States)

    Nagaviroj, Kittiphon; Anothaisintawee, Thunyarat

    2017-06-01

    Many terminally ill patients would prefer to stay and die in their own homes, but unfortunately, some may not be able to do so. Although there are many factors associated with successful home deaths, receiving palliative home visits from the multidisciplinary care teams is one of the key factors that enable patients to die at home. Our study was aimed to find whether there was any association between our palliative home care program and home death. A retrospective study was conducted in the Department of Family Medicine at Ramathibodi Hospital between January 2012 and May 2014. All of the patients who were referred to multidisciplinary palliative care teams were included. The data set comprised of patient's profile, disease status, functional status, patient's symptoms, preferred place of death, frequency of home visits, types of team interventions, and patient's actual place of death. Multiple logistic regression was applied in order to determine the association between the variables and the probability of dying at home. A total of 142 patients were included into the study. At the end of the study, 50 (35.2%) patients died at home and 92 (64.8%) patients died in the hospital. The multivariate logistic regression analysis demonstrated a strong association between multidisciplinary home care and home death (odds ratio 6.57, 95% confidence interval [CI] 2.48-17.38). Palliative home care was a significant factor enabling patients who want to die at home. We encourage health policy makers to promote the development of community-based palliative care programs in Thailand.

  12. Frequency and magnetic field mapping of magnetoelastic spin pumping in high overtone bulk acoustic wave resonator

    Directory of Open Access Journals (Sweden)

    N. I. Polzikova

    2018-05-01

    Full Text Available We report on the first observation of microvolt-scale inverse spin Hall effect (ISHE dc voltage driven by an acoustic spin pumping (ASP in a bulk acoustic wave (BAW resonator formed by a Al-ZnO-Al-YIG(1-GGG-YIG(2-Pt structure. When 2 mW power is applied to an Al-ZnO-Al transducer, the voltage VISHE ∼ 4 μV in the Pt film is observed as a result of resonant ASP from YIG(2 to Pt in the area ∼ 170 μm. The results of frequency and magnetic field mapping of VISHE(f,H together with reflectivity of the resonator show an obvious agreement between the positions of the voltage maxima and BAW resonance frequencies fn(H on the (f, H plane. At the same time a significant asymmetry of the VISHE(fn(H value in reference to the magnetoelastic resonance (MER line fMER(H position is revealed, which is explained by asymmetry of the magnetoelastic waves dispersion law.

  13. Magnon Valve Effect between Two Magnetic Insulators

    Science.gov (United States)

    Wu, H.; Huang, L.; Fang, C.; Yang, B. S.; Wan, C. H.; Yu, G. Q.; Feng, J. F.; Wei, H. X.; Han, X. F.

    2018-03-01

    The key physics of the spin valve involves spin-polarized conduction electrons propagating between two magnetic layers such that the device conductance is controlled by the relative magnetization orientation of two magnetic layers. Here, we report the effect of a magnon valve which is made of two ferromagnetic insulators (YIG) separated by a nonmagnetic spacer layer (Au). When a thermal gradient is applied perpendicular to the layers, the inverse spin Hall voltage output detected by a Pt bar placed on top of the magnon valve depends on the relative orientation of the magnetization of two YIG layers, indicating the magnon current induced by the spin Seebeck effect at one layer affects the magnon current in the other layer separated by Au. We interpret the magnon valve effect by the angular momentum conversion and propagation between magnons in two YIG layers and conduction electrons in the Au layer. The temperature dependence of the magnon valve ratio shows approximately a power law, supporting the above magnon-electron spin conversion mechanism. This work opens a new class of valve structures beyond the conventional spin valves.

  14. Biometrics for home networks security

    KAUST Repository

    Ansari, Imran Shafique; Ahmad, Qutbuddin S.

    2009-01-01

    Hacking crimes committed to the home networks are increasing. Advanced network protection is not always possible for the home networks. In this paper we will study the ability of using biometric systems for authentication in home networks. ©2009

  15. Biometrics for home networks security

    KAUST Repository

    Ansari, Imran Shafique

    2009-01-01

    Hacking crimes committed to the home networks are increasing. Advanced network protection is not always possible for the home networks. In this paper we will study the ability of using biometric systems for authentication in home networks. ©2009 IEEE.

  16. The Future of Home Health project: developing the framework for health care at home.

    Science.gov (United States)

    Lee, Teresa; Schiller, Jennifer

    2015-02-01

    In addition to providing high-quality care to vulnerable patient populations, home healthcare offers the least costly option for patients and the healthcare system, particularly in postacute care. As the baby boom generation ages, policymakers are expressing concerns about rising costs, variation in home healthcare service use, and program integrity. The Alliance for Home Health Quality and Innovation seeks to develop a research-based strategic framework for the future of home healthcare for older Americans and those with disabilities. This article describes the initiative and invites readers to provide comments and suggestions.

  17. A Home Away from Home

    DEFF Research Database (Denmark)

    McIlvenny, Paul

    2008-01-01

    The House of Tiny Tearaways (HTT) first appeared on British television in May 2005. Over a six-day period, three families are invited to reside in a specially designed house together with a resident clinical psychologist. The house is to be “a home away from home” for the resident families...... in order to analyze excerpts from the program and to explore how the affordances and constraints of the specially designed house—its architecture and spatial configuration, as well as the surveillance technology embedded within its walls—are assembled within particular familial activities, and how...... the relationships between family members are reshaped as a result. The analysis focuses on several key phenomena: 1) practices of video observation in relation to the domestic sphere; 2) use of inscription devices, such as video displays, to capture and visualize behavior and action in the “home;” 3) practicing...

  18. The Future of Home Health Care

    Science.gov (United States)

    Landers, Steven; Madigan, Elizabeth; Leff, Bruce; Rosati, Robert J.; McCann, Barbara A.; Hornbake, Rodney; MacMillan, Richard; Jones, Kate; Bowles, Kathryn; Dowding, Dawn; Lee, Teresa; Moorhead, Tracey; Rodriguez, Sally; Breese, Erica

    2016-01-01

    The Future of Home Health project sought to support transformation of home health and home-based care to meet the needs of patients in the evolving U.S. health care system. Interviews with key thought leaders and stakeholders resulted in key themes about the future of home health care. By synthesizing this qualitative research, a literature review, case studies, and the themes from a 2014 Institute of Medicine and National Research Council workshop on “The Future of Home Health Care,” the authors articulate a vision for home-based care and recommend a bold framework for the Medicare-certified home health agency of the future. The authors also identify challenges and recommendations for achievement of this framework. PMID:27746670

  19. Home care services for sick children

    DEFF Research Database (Denmark)

    Castor, Charlotte; Hallström, Inger; Hansson, Eva Helena

    2017-01-01

    as challenging for healthcare professionals in home care services used to providing care predominately for adults. DESIGN: An inductive qualitative design. METHOD: Seven focus group interviews were performed with 36 healthcare professionals from multidisciplinary home care services. Data were analysed stepwise......AIMS AND OBJECTIVES: To explore healthcare professionals' conceptions of caring for sick children in home care services. BACKGROUND: Families often prefer home care to hospital care, and the number of home care services for children is increasing. Caring for children at home has been recognised...... using a phenomenographic analysis. RESULTS: Three description categories emerged: "A challenging opportunity", "A child perspective", and "Re-organise in accordance with new prerequisites." Providing home care services for children was conceived to evoke both professional and personal challenges...

  20. Acquiring taste in home economics?

    DEFF Research Database (Denmark)

    Stenbak Larsen, Christian

    Objective: To explore how home economics was taught in Denmark before the recent Danish school reform, which also revised the objectives and content of home economics, naming it Food Knowledge (Madkundskab) Methods: Participant observation was done in home economic lessons in two case schools...... appreciated by the group of boys, and others again learned to stick with their idiosyncrasies when pressured by the teacher. Conclusions: Children were acquiring taste in the home economic lessons, but not only the kind of tastes that the teacher had planned for. This leads to reflections on the very complex...... process of taste acquiring and to a call for further research into taste acquiring in complex real life contexts as home economics lessons....

  1. Design and simulation of self-biased circulators in the ultra high frequency band

    International Nuclear Information System (INIS)

    Wang Jianwei; Geiler, Anton; Mistry, Perhaad; Kaeli, David R.; Harris, Vincent G.; Vittoria, Carmine

    2012-01-01

    Theoretical models were developed to design self-biased Y-junction circulators operating at ultra high frequency (UHF). The proposed circulator designs consist of insulating nanowires of yttrium iron garnet (YIG) embedded in high permittivity barium–strontium titanate (BSTO) substrates. A design with as many as 10 5 or greater wires may be considered in its entirety to determine the electromagnetic scattering S-parameters of a circulator design, thus helping to mitigate the computational limitations of the available finite element method (FEM) tools. The approach seeks to represent the nanowires and the BSTO substrate by an equivalent medium with effective properties inclusive of the average saturation magnetization, dynamic demagnetizing fields, and permittivity. The effective medium approach was validated in comparison with the FEM models. Using the proposed approach, a self-biased junction circulator consisting of YIG nanowires embedded in a BSTO substrate was designed and simulated in which the center frequency insertion loss was calculated to be as low as 0.16 dB with isolation of −42.3 dB at 1 GHz. The 20 dB bandwidth was calculated to be 50 MHz. These results suggest that practical self-biased circulators at the UHF band are feasible. - Highlights: ► Presented a self-biased Y-junction circulator topology on composite substrate with YIG nanowires and high permittivity BSTO. ► Developed an equivalent model to characterize the composite substrate. ► Designed a self-biased junction circulator consisting of YIG nanowires embedded in a BSTO substrate at 1 GHz.

  2. Strongly extended diffusion length for the nonequilibrium magnons in Y3F e5O12 by photoexcitation

    Science.gov (United States)

    Wang, S. H.; Li, G.; Guo, E. J.; Zhao, Y.; Wang, J. Y.; Zou, L. K.; Yan, H.; Cai, J. W.; Zhang, Z. T.; Wang, M.; Tian, Y. Y.; Zheng, X. L.; Sun, J. R.; Jin, K. X.

    2018-05-01

    Y3F e5O12 (YIG) is known for its long magnon diffusion length. Although it has the known lowest damping rate, an even longer diffusion distance is still highly desired since it may lead to a much more efficient information transmission and processing. While most of previous works focused on the generation and detection of magnons in YIG, here we demonstrate how to depress the damping rate during the diffusion of magnon. By selectively exciting the spin state transition of the Fe ions in YIG, we successfully increase magnon diffusion length by one order of magnitude, i.e., from the previous reported ˜10 μm up to ˜156 μm (for the sample prepared by liquid phase epitaxy) and ˜180 μm (for the sample prepared by pulsed laser deposition) at room temperature. The diffusion length, determined by nonlocal geometry, is ˜30 μm for the magnons induced by visible light and above 150 μm for the laser of 980 nm. In addition to thermal gradient, light excitation affects the electron configuration of the F e3 + ion in YIG. Long-wavelength laser is more effective since it causes a transition of the F e3 + ions in Fe O6 octahedron from a high spin to a low spin state and thus causes a magnon softening which favors a long-distance diffusion. The present work paves the way toward an efficient tuning of magnon transport which is crucially important for magnon spintronics.

  3. Home Learning, Technology, and Tomorrow's Workplace.

    Science.gov (United States)

    Rieseberg, Rhonda L.

    1995-01-01

    Discusses characteristics and trends of home schools and workplaces. Use of computers and computer applications (CD-ROMS, interactive software, and networking) in home schooling provides a compatible environment for future home-based businesses and telecommuting trends. Sidebars include information on home schools on line; standardized test…

  4. Community Nursing Home (CNH)

    Data.gov (United States)

    Department of Veterans Affairs — The Community Nursing Home (CNH) database contains a list of all Community Nursing Home facilities under local contract to Veterans Health Administration (VHA). CNH...

  5. Optimal taxation with home production

    OpenAIRE

    Olovsson, Conny

    2014-01-01

    Optimal taxes for Europe and the U.S. are derived in a realistically calibrated model in which agents buy consumption goods and services and use home capital and labor to produce household services. The optimal tax rate on services is substantially lower than the tax rate on goods. Specifically, the planner cannot tax home production directly and instead lowers the tax rate on market services to increase the relative price of home production. The optimal tax rate on the return to home capital...

  6. Home Automation and Security System

    OpenAIRE

    Surinder Kaur,; Rashmi Singh; Neha Khairwal; Pratyk Jain

    2016-01-01

    Easy Home or Home automation plays a very important role in modern era because of its flexibility in using it at different places with high precision which will save money and time by decreasing human hard work. Prime focus of this technology is to control the household equipment’s like light, fan, door, AC etc. automatically. This research paper has detailed information on Home Automation and Security System using Arduino, GSM and how we can control home appliances using Android application....

  7. Nursing home-acquired pneumonia, dysphagia and associated diseases in nursing home residents: A retrospective, cross-sectional study

    NARCIS (Netherlands)

    Hollaar, V.R.Y.; Putten, G.J. van der; Maarel-Wierink, C.D. van der; Bronkhorst, E.M.; Swart, B.J.M. de; Baat, C. de; Creugers, N.H.J.

    2017-01-01

    BACKGROUND: Nursing home-acquired pneumonia (NHAP) is a common infection among nursing home residents. There is also a high prevalence of dysphagia in nursing home residents and they suffer more often from comorbidity and multimorbidity. This puts nursing home residents at higher risk of (mortality

  8. 78 FR 46421 - Proposed Information Collection (Per Diem for Nursing Home Care of Veterans in State Homes; Per...

    Science.gov (United States)

    2013-07-31

    ... State homes providing nursing home and adult day health services care to Veterans. VA requires... Diem for Nursing Home Care of Veterans in State Homes; Per Diem for Adult Day Care of Veterans in State... information needed to ensure that nursing home and adult day health care facilities are providing high quality...

  9. High-resolution crystal structure reveals a HEPN domain at the C-terminal region of S. cerevisiae RNA endonuclease Swt1

    International Nuclear Information System (INIS)

    Peng, Shuxia; Zhou, Ke; Wang, Wenjia; Gao, Zengqiang; Dong, Yuhui; Liu, Quansheng

    2014-01-01

    Highlights: • Crystal structure of the C-terminal (CT) domain of Swt1 was determined at 2.3 Å. • Structure of the CT domain was identified as HEPN domain superfamily member. • Low-resolution envelope of Swt1 full-length in solution was analyzed by SAXS. • The middle and CT domains gave good fit to SAXS structural model. - Abstract: Swt1 is an RNA endonuclease that plays an important role in quality control of nuclear messenger ribonucleoprotein particles (mRNPs) in eukaryotes; however, its structural details remain to be elucidated. Here, we report the crystal structure of the C-terminal (CT) domain of Swt1 from Saccharomyces cerevisiae, which shares common characteristics of higher eukaryotes and prokaryotes nucleotide binding (HEPN) domain superfamily. To study in detail the full-length protein structure, we analyzed the low-resolution architecture of Swt1 in solution using small angle X-ray scattering (SAXS) method. Both the CT domain and middle domain exhibited a good fit upon superimposing onto the molecular envelope of Swt1. Our study provides the necessary structural information for detailed analysis of the functional role of Swt1, and its importance in the process of nuclear mRNP surveillance

  10. Rate-determining Step of Flap Endonuclease 1 (FEN1) Reflects a Kinetic Bias against Long Flaps and Trinucleotide Repeat Sequences.

    Science.gov (United States)

    Tarantino, Mary E; Bilotti, Katharina; Huang, Ji; Delaney, Sarah

    2015-08-21

    Flap endonuclease 1 (FEN1) is a structure-specific nuclease responsible for removing 5'-flaps formed during Okazaki fragment maturation and long patch base excision repair. In this work, we use rapid quench flow techniques to examine the rates of 5'-flap removal on DNA substrates of varying length and sequence. Of particular interest are flaps containing trinucleotide repeats (TNR), which have been proposed to affect FEN1 activity and cause genetic instability. We report that FEN1 processes substrates containing flaps of 30 nucleotides or fewer at comparable single-turnover rates. However, for flaps longer than 30 nucleotides, FEN1 kinetically discriminates substrates based on flap length and flap sequence. In particular, FEN1 removes flaps containing TNR sequences at a rate slower than mixed sequence flaps of the same length. Furthermore, multiple-turnover kinetic analysis reveals that the rate-determining step of FEN1 switches as a function of flap length from product release to chemistry (or a step prior to chemistry). These results provide a kinetic perspective on the role of FEN1 in DNA replication and repair and contribute to our understanding of FEN1 in mediating genetic instability of TNR sequences. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  11. Ostomy Home Skills Program

    Medline Plus

    Full Text Available ... ACS Careers at ACS About ACS Career Types Working at ACS ... Education Patients and Family Skills Programs Ostomy Home Skills Program Ostomy Home Skills Program Adult Ostomy ...

  12. Home advantage in professional tennis

    NARCIS (Netherlands)

    Koning, Ruud H.

    2011-01-01

    Home advantage is a pervasive phenomenon in sport. It has been established in team sports such as basketball, baseball, American football, and European soccer. Attention to home advantage in individual sports has so far been limited. The aim of this study was to examine home advantage in

  13. Modular Zero Energy. BrightBuilt Home

    Energy Technology Data Exchange (ETDEWEB)

    Aldrich, Robb [Consortium for Advanced Residential Buildings (CARB), Norwalk, CT (United States). Steven Winters Associates, Inc.; Butterfield, Karla [Consortium for Advanced Residential Buildings (CARB), Norwalk, CT (United States). Steven Winters Associates, Inc.

    2016-03-01

    Kaplan Thompson Architects (KTA) has specialized in sustainable, energy-efficient buildings, and they have designed several custom, zero-energy homes in New England. These zero-energy projects have generally been high-end, custom homes with budgets that could accommodate advanced energy systems. In an attempt to make zero energy homes more affordable and accessible to a larger demographic, KTA explored modular construction as way to provide high-quality homes at lower costs. In the mid-2013, KTA formalized this concept when they launched BrightBuilt Home (BBH). The BBH mission is to offer a line of architect-designed, high-performance homes that are priced to offer substantial savings off the lifetime cost of a typical home and can be delivered in less time. For the past two years, CARB has worked with BBH and Keiser Homes (the primary modular manufacturer for BBH) to discuss challenges related to wall systems, HVAC, and quality control. In Spring of 2014, CARB and BBH began looking in detail on a home to be built in Lincolnville, ME by Black Bros. Builders. This report details the solution package specified for this modular plan and the challenges that arose during the project.

  14. Predictors of home death of home palliative cancer care patients: a cross-sectional nationwide survey.

    Science.gov (United States)

    Fukui, Sakiko; Fujita, Junko; Tsujimura, Mayuko; Sumikawa, Yuka; Hayashi, Yayoi

    2011-11-01

    To identify factors influencing the place of death among home palliative cancer care patients, focusing on the role of nurses in terms of pre- and post-discharge from hospital to home care settings. A cross-sectional nationwide questionnaire survey was conducted at 1000 randomly selected homecare agencies in Japan. The questionnaires were completed by primary community nurses of home palliative patients just after their discharge. A total of 568 responses were analyzed (effective response rate, 69%). Multivariate logistic regression analysis revealed the following independent factors of place of death among those patients: desire for home death at referral by both patient and family caregiver; caregiver relationship to patient as daughter or daughter-in-law; totally bedridden functional status of patient; patient not suffering from depression and/or anxiety at referral; patients and caregivers duly informed about the dying process/death in detail, as well as instructed by community nurses about pain management and how to treat/prevent bedsores in home care settings. This study demonstrated the importance of both the hospital and community nurses' role in increasing the patients' chance of dying at home. Hospital nurses should support early transfer to home palliative care according to their assessment of the desire of patient/family caregiver for home death, the patients' clinical status, and caregivers' ability to provide patient care at home. Community nurses should inform patients/family caregiver in detail about the dying process/death just after discharge, relieve patient pain, treat/prevent bedsores, and instruct family caregivers on their symptom control. Crown Copyright © 2011. Published by Elsevier Ltd. All rights reserved.

  15. Vocational Home Economics Curriculum Guide for Occupational Home and Institutional Services.

    Science.gov (United States)

    Dewald, Margaret R.

    The training program outlined in this guide focuses upon the development of students for gainful employment through a two-year course of study in home and institutional services. Instructional topics are provided in nine areas: orientation to home and institutional services, cleaning supplies and equipment, cleaning surfaces, cleaning clothes and…

  16. 77 FR 45719 - Proposed Information Collection (Per Diem for Nursing Home Care of Veterans in State Homes; Per...

    Science.gov (United States)

    2012-08-01

    ... per diem to State homes providing nursing home and adult day health services care to Veterans. VA... Diem for Nursing Home Care of Veterans in State Homes; Per Diem for Adult Day Care of Veterans in State... information needed to ensure that nursing home and adult day health care facilities are providing high quality...

  17. Nursing Home Quality Initiative

    Data.gov (United States)

    U.S. Department of Health & Human Services — This Nursing Home Quality Initiative (NHQI) website provides consumer and provider information regarding the quality of care in nursing homes. NHQI discusses quality...

  18. 7 CFR 1755.509 - Mobile homes.

    Science.gov (United States)

    2010-01-01

    ... homes, motor homes, truck campers, travel trailers, and all forms of recreational vehicles. Customer... ® for a mobile home depends on how the electric power is installed at the mobile home and it can involve considerable judgment on the part of the telecommunications installer. The National Electrical Code ® and NEC...

  19. The HOME Inventory and Family Demographics.

    Science.gov (United States)

    Bradley, Robert H.; Caldwell, Bettye M.

    1984-01-01

    Examines the relation between the Home Observation for Measurement of Environment (HOME) Inventory and sex, race, socioeconomic status, the amount of crowding in the home, and birth order. Performs multivariate analysis of covariance on an intact family sample using HOME subscales as criterion measures and status and structural variables as…

  20. Custodial Homes, Therapeutic Homes, and Parental Acceptance: Parental Experiences of Autism in Kerala, India and Atlanta, GA USA.

    Science.gov (United States)

    Sarrett, Jennifer C

    2015-06-01

    The home is a critical place to learn about cultural values of childhood disability, including autism and intellectual disabilities. The current article describes how the introduction of autism into a home and the availability of intervention options change the structure and meaning of a home and reflect parental acceptance of a child's autistic traits. Using ethnographic data from Kerala, India and Atlanta, GA USA, a description of two types of homes are developed: the custodial home, which is primarily focused on caring for basic needs, and the therapeutic home, which is focused on changing a child's autistic traits. The type of home environment is respondent to cultural practices of child rearing in the home and influences daily activities, management, and care in the home. Further, these homes differ in parental acceptance of their autistic children's disabilities, which is critical to understand when engaging in international work related to autism and intellectual disability. It is proposed that parental acceptance can be fostered through the use of neurodiverse notions that encourage autism acceptance.

  1. Home e-health system integration in the Smart Home through a common media server.

    Science.gov (United States)

    Pau, I; Seoane, F; Lindecrantz, K; Valero, M A; Carracedo, J

    2009-01-01

    Home e-health systems and services are revealed as one of the most important challenges to promote Quality of Life related to Health in the Information Society. Leading companies have worked on e-health systems although the majority of them are addressed to hospital or primary care settings. The solution detailed in this paper offers a personal health system to be integrated with Smart Home services platform to support home based e-care. Thus, the home e-health system and architecture detailed in this research work is ready to supply a seamless personal care solution both from the biomedical data analysis, service provision, security guarantee and information management s point of view. The solution is ready to be integrated within the Accessible Digital Home, a living lab managed by Universidad Politécnica de Madrid for R&D activities.

  2. Ensuring Quality Nursing Home Care

    Science.gov (United States)

    ... cases, you can also call the Department of Health. Nursing homes are required to post information on how you ... nursing homes in your area, go to Medicare’s Nursing Home Compare website at ... information is not intended to diagnose health problems or to take the place of medical ...

  3. Strong Interlayer Magnon-Magnon Coupling in Magnetic Metal-Insulator Hybrid Nanostructures

    Science.gov (United States)

    Chen, Jilei; Liu, Chuanpu; Liu, Tao; Xiao, Yang; Xia, Ke; Bauer, Gerrit E. W.; Wu, Mingzhong; Yu, Haiming

    2018-05-01

    We observe strong interlayer magnon-magnon coupling in an on-chip nanomagnonic device at room temperature. Ferromagnetic nanowire arrays are integrated on a 20-nm-thick yttrium iron garnet (YIG) thin film strip. Large anticrossing gaps up to 1.58 GHz are observed between the ferromagnetic resonance of the nanowires and the in-plane standing spin waves of the YIG film. Control experiments and simulations reveal that both the interlayer exchange coupling and the dynamical dipolar coupling contribute to the observed anticrossings. The coupling strength is tunable by the magnetic configuration, allowing the coherent control of magnonic devices.

  4. Superconductivity induced by interfacial coupling to magnons

    Science.gov (United States)

    Rohling, Niklas; Fjærbu, Eirik Løhaugen; Brataas, Arne

    2018-03-01

    We consider a thin normal metal sandwiched between two ferromagnetic insulators. At the interfaces, the exchange coupling causes electrons within the metal to interact with magnons in the insulators. This electron-magnon interaction induces electron-electron interactions, which in turn can result in p -wave superconductivity. We solve the gap equation numerically and estimate the critical temperature. In yttrium iron garnet (YIG)-Au-YIG trilayers, superconductivity sets in at temperatures somewhere in the interval between 1 and 10 K. EuO-Au-EuO trilayers require a lower temperature, in the range from 0.01 to 1 K.

  5. The 'Cancer Home-Life Intervention'

    DEFF Research Database (Denmark)

    Pilegaard, Marc Sampedro; la Cour, Karen; Gregersen Oestergaard, Lisa

    2018-01-01

    , occupational therapy-based and adaptive programme for people with advanced cancer targeting the performance of their prioritised everyday activities. SETTING/PARTICIPANTS: Home-living adults diagnosed with advanced cancer experiencing functional limitations were recruited from two Danish hospitals. They were......BACKGROUND: People with advanced cancer face difficulties with their everyday activities at home that may reduce their health-related quality of life. To address these difficulties, we developed the 'Cancer Home-Life Intervention'. AIM: To evaluate the efficacy of the 'Cancer Home Life......-Intervention' compared with usual care with regard to patients' performance of, and participation in, everyday activities, and their health-related quality of life. DESIGN AND INTERVENTION: A randomised controlled trial ( ClinicalTrials.gov NCT02356627). The 'Cancer Home-Life Intervention' is a brief, tailored...

  6. DNA and Protein Requirements for Substrate Conformational Changes Necessary for Human Flap Endonuclease-1-catalyzed Reaction*

    Science.gov (United States)

    Algasaier, Sana I.; Exell, Jack C.; Bennet, Ian A.; Thompson, Mark J.; Gotham, Victoria J. B.; Shaw, Steven J.; Craggs, Timothy D.; Finger, L. David; Grasby, Jane A.

    2016-01-01

    Human flap endonuclease-1 (hFEN1) catalyzes the essential removal of single-stranded flaps arising at DNA junctions during replication and repair processes. hFEN1 biological function must be precisely controlled, and consequently, the protein relies on a combination of protein and substrate conformational changes as a prerequisite for reaction. These include substrate bending at the duplex-duplex junction and transfer of unpaired reacting duplex end into the active site. When present, 5′-flaps are thought to thread under the helical cap, limiting reaction to flaps with free 5′-termini in vivo. Here we monitored DNA bending by FRET and DNA unpairing using 2-aminopurine exciton pair CD to determine the DNA and protein requirements for these substrate conformational changes. Binding of DNA to hFEN1 in a bent conformation occurred independently of 5′-flap accommodation and did not require active site metal ions or the presence of conserved active site residues. More stringent requirements exist for transfer of the substrate to the active site. Placement of the scissile phosphate diester in the active site required the presence of divalent metal ions, a free 5′-flap (if present), a Watson-Crick base pair at the terminus of the reacting duplex, and the intact secondary structure of the enzyme helical cap. Optimal positioning of the scissile phosphate additionally required active site conserved residues Tyr40, Asp181, and Arg100 and a reacting duplex 5′-phosphate. These studies suggest a FEN1 reaction mechanism where junctions are bound and 5′-flaps are threaded (when present), and finally the substrate is transferred onto active site metals initiating cleavage. PMID:26884332

  7. The first multicenter, randomized, controlled trial of home telemonitoring for Japanese patients with heart failure: home telemonitoring study for patients with heart failure (HOMES-HF).

    Science.gov (United States)

    Kotooka, Norihiko; Kitakaze, Masafumi; Nagashima, Kengo; Asaka, Machiko; Kinugasa, Yoshiharu; Nochioka, Kotaro; Mizuno, Atsushi; Nagatomo, Daisuke; Mine, Daigo; Yamada, Yoko; Kuratomi, Akiko; Okada, Norihiro; Fujimatsu, Daisuke; Kuwahata, So; Toyoda, Shigeru; Hirotani, Shin-Ichi; Komori, Takahiro; Eguchi, Kazuo; Kario, Kazuomi; Inomata, Takayuki; Sugi, Kaoru; Yamamoto, Kazuhiro; Tsutsui, Hiroyuki; Masuyama, Tohru; Shimokawa, Hiroaki; Momomura, Shin-Ichi; Seino, Yoshihiko; Sato, Yasunori; Inoue, Teruo; Node, Koichi

    2018-02-15

    Home telemonitoring is becoming more important to home medical care for patients with heart failure. Since there are no data on home telemonitoring for Japanese patients with heart failure, we investigated its effect on cardiovascular outcomes. The HOMES-HF study was the first multicenter, open-label, randomized, controlled trial (RCT) to elucidate the effectiveness of home telemonitoring of physiological data, such as body weight, blood pressure, and pulse rate, for Japanese patients with heart failure (UMIN Clinical Trials Registry 000006839). The primary end-point was a composite of all-cause death or rehospitalization due to worsening heart failure. We analyzed 181 recently hospitalized patients with heart failure who were randomly assigned to a telemonitoring group (n = 90) or a usual care group (n = 91). The mean follow-up period was 15 (range 0-31) months. There was no statistically significant difference in the primary end-point between groups [hazard ratio (HR), 0.95; 95% confidence interval (CI), 0.548-1.648; p = 0.572]. Home telemonitoring for Japanese patients with heart failure was feasible; however, beneficial effects in addition to those of usual care were not demonstrated. Further investigation of more patients with severe heart failure, participation of home medical care providers, and use of a more integrated home telemonitoring system emphasizing communication as well as monitoring of symptoms and physiological data are required.

  8. Genetic Mediation of the Home Environment during Infancy: A Sibling Adoption Study of the HOME.

    Science.gov (United States)

    Braungart, Julia M.; And Others

    1992-01-01

    The home environment of nonadoptive and adoptive sibling pairs was assessed using the Home Observation for Measurement of the Environment when each sibling was one and two years of age. Correlations between home environment scores for nonadoptive siblings were greater than those for adoptive siblings. (BC)

  9. Randomized trial of the ForeseeHome monitoring device for early detection of neovascular age-related macular degeneration. The HOme Monitoring of the Eye (HOME) study design - HOME Study report number 1.

    Science.gov (United States)

    Chew, Emily Y; Clemons, Traci E; Bressler, Susan B; Elman, Michael J; Danis, Ronald P; Domalpally, Amitha; Heier, Jeffrey S; Kim, Judy E; Garfinkel, Richard A

    2014-03-01

    To evaluate the effects of a home-monitoring device with tele-monitoring compared with standard care in detection of progression to choroidal neovascularization (CNV) associated with age-related macular degeneration (AMD), the leading cause of blindness in the US. Participants, aged 55 to 90 years, at high risk of developing CNV associated with AMD were recruited to the HOme Monitoring of Eye (HOME) Study, an unmasked, multi-center, randomized trial of the ForeseeHome (FH) device plus standard care vs. standard care alone. The FH device utilizes preferential hyperacuity perimetry and tele-monitoring to detect changes in vision function associated with development of CNV, potentially prior to symptom and visual acuity loss. After establishing baseline measurements, subsequent changes on follow-up are detected by the device, causing the monitoring center to alert the clinical center to recall participants for an exam. Standard care consists of instructions for self-monitoring visual changes with subsequent self-report to the clinical center. The primary objective of this study is to determine whether home monitoring plus standard care in comparison with standard care alone, results in earlier detection of incident CNV with better present visual acuity. The primary outcome is the decline in visual acuity at CNV diagnosis from baseline. Detection of CNV prior to substantial vision loss is critical as vision outcome following anti-angiogenic therapy is dependent on the visual acuity at initiation of treatment. HOME Study is the first large scale study to test the use of home tele-monitoring system in the management of AMD patients. Published by Elsevier Inc.

  10. Factors related to the provision of home-based end-of-life care among home-care nursing, home help, and care management agencies in Japan.

    Science.gov (United States)

    Igarashi, Ayumi; Kurinobu, Takeshi; Ko, Ayako; Okamoto, Yuko; Matsuura, Shino; Feng, Mei; Yamamoto-Mitani, Noriko

    2015-09-12

    To promote home death, it is necessary to clarify the institutional barriers to conducting end-of-life (EOL) care and consider strategies to deal with this process. This study aims to clarify institution-related factors associated with the provision of home-based EOL care cases, and to compare them among three different types of home-care agencies. We administered a cross-sectional survey throughout Japan to investigate the number and characteristics of EOL cases of home-care nursing (HN), home-help (HH) and care management (CM) agencies. Bivariate and multivariate analyses were performed for each type of agency to examine factors related to the provision of EOL care. 378 HN agencies, 274 HH agencies, and 452 CM agencies responded to the distributed questionnaire. HN agencies had on average 2.1 (SD = 4.0; range 0-60) home-based EOL cases in the last 3 months, while HH agencies had 0.9 (SD = 1.3; range 0-7) and CM agencies had 1.5 (SD = 2.2; range 0-18) in the last 6 months. In a multivariable analysis of HN agencies, a large number of staff (OR: 1.52; p EOL care; in HH agencies, accepting EOL clients in the agency (OR: 3.29; p EOL care; in CM agencies, the number of staff (OR: 1.21; p = 0.037), the number of collaborating HH agencies (OR: 1.07; p = 0.032), and whether home-care nurses and home helpers visit clients together (OR: 1.89; p = 0.007) were positively associated with the provision of EOL care. The agency's size and the inter-agency collaborative system seemed most important among HN agencies and CM agencies, while institutional preparedness for EOL was most important for HH agencies. These findings represent important new information for targeting different effective strategies in the promotion of home-based EOL care, depending on the agency type.

  11. HOME CARE NURSES’ ROLES IN ENHANCING QUALITY OF NURSING CARE FOR PATIENTS AT HOME: A PHENOMENOLOGICAL STUDY

    OpenAIRE

    Titan Ligita

    2017-01-01

    Background: Provision of health care service at home is one of the advanced forms of care for patients being discharged from hospitalization. Little is known about the experience of nurses providing home care services through a nursing home-care model especially in Indonesian context. Objective: This study aims to explore the experience in order to increase understanding on the form of home care provision, and consequently the nurses may understand the form of home care globally. Metho...

  12. The incidence of depression and its risk factors in Dutch nursing homes and residential care homes.

    Science.gov (United States)

    Boorsma, Marijke; Joling, Karlijn; Dussel, Martine; Ribbe, Miel; Frijters, Dinnus; van Marwijk, Harm W J; Nijpels, Giel; van Hout, Hein

    2012-11-01

    Although it is known that depression is highly prevalent in institutionalized older adults, little is known about its incidence and risk factors in nursing homes and residential care homes. The aim of this study was to investigate and compare the incidence and associated risk factors for depression in Dutch nursing homes and residential care homes. Data on depression were extracted from the Vrije Universiteit naturalistic cohort on routine care monitoring with the Minimum Data Set of the Resident Assessment Instrument. A total of 1,324 residents in six nursing homes and 1,723 residents in 23 residential care homes with an average follow-up of 1.2 years. Depression was defined as a clinical diagnosis according to Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, criteria and the use of antidepressants. Residents with prevalent depression at baseline were excluded. The incidence rate was 13.6 per 100 person years in the nursing homes and 10.2 per 100 person years in the residential care homes. The independent risk factors for in-home depression for residents in nursing homes included dementia (OR: 1.7; 95% CI: 1.02-2.95) and a score of 3 or more on the Depression Rating Scale (odds ratio [OR]: 2.1; 95% confidence interval [CI]: 1.23-3.70). A protective effect was seen on the use of a hearing aid (OR: 0.3; 95% CI: 0.12-0.80). In the residential care homes, being male (OR: 2.1; 95% CI: 1.27-3.30), having cancer (OR: 2.9; 95% CI: 1.64-4.95), and a score of 2 or higher on the Cognitive Performance Scale (OR: 1.5; 95% CI: 1.05-2.22) increased the risk to develop depression. Age greater than 85 years (OR: 0.5; 95% CI: 0.31-0.67) and hearing impairment (OR: 0.8; 95% CI: 0.60-1.00) appeared to be protective. The incidence rate for depression in residents of Dutch nursing homes and residential care homes was high and the associated risk factors found may have important implications for staff. 2012 American Association for Geriatric Psychiatry

  13. Ostomy Home Skills Program

    Medline Plus

    Full Text Available ... Careers at ACS Careers at ACS About ACS Career Types Working at ACS ... American College of Surgeons Education Patients and Family Skills Programs Ostomy Home Skills Program Ostomy Home Skills ...

  14. Cardiomyocyte hypertrophy induced by Endonuclease G deficiency requires reactive oxygen radicals accumulation and is inhibitable by the micropeptide humanin.

    Science.gov (United States)

    Blasco, Natividad; Cámara, Yolanda; Núñez, Estefanía; Beà, Aida; Barés, Gisel; Forné, Carles; Ruíz-Meana, Marisol; Girón, Cristina; Barba, Ignasi; García-Arumí, Elena; García-Dorado, David; Vázquez, Jesús; Martí, Ramon; Llovera, Marta; Sanchis, Daniel

    2018-06-01

    The endonuclease G gene (Endog), which codes for a mitochondrial nuclease, was identified as a determinant of cardiac hypertrophy. How ENDOG controls cardiomyocyte growth is still unknown. Thus, we aimed at finding the link between ENDOG activity and cardiomyocyte growth. Endog deficiency induced reactive oxygen species (ROS) accumulation and abnormal growth in neonatal rodent cardiomyocytes, altering the AKT-GSK3β and Class-II histone deacethylases (HDAC) signal transduction pathways. These effects were blocked by ROS scavengers. Lack of ENDOG reduced mitochondrial DNA (mtDNA) replication independently of ROS accumulation. Because mtDNA encodes several subunits of the mitochondrial electron transport chain, whose activity is an important source of cellular ROS, we investigated whether Endog deficiency compromised the expression and activity of the respiratory chain complexes but found no changes in these parameters nor in ATP content. MtDNA also codes for humanin, a micropeptide with possible metabolic functions. Nanomolar concentrations of synthetic humanin restored normal ROS levels and cell size in Endog-deficient cardiomyocytes. These results support the involvement of redox signaling in the control of cardiomyocyte growth by ENDOG and suggest a pathway relating mtDNA content to the regulation of cell growth probably involving humanin, which prevents reactive oxygen radicals accumulation and hypertrophy induced by Endog deficiency. Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.

  15. Inhibitors of nuclease and redox activity of apurinic/apyrimidinic endonuclease 1/redox effector factor 1 (APE1/Ref-1).

    Science.gov (United States)

    Laev, Sergey S; Salakhutdinov, Nariman F; Lavrik, Olga I

    2017-05-01

    Human apurinic/apyrimidinic endonuclease 1/redox effector factor 1 (APE1/Ref-1) is a multifunctional protein which is essential in the base excision repair (BER) pathway of DNA lesions caused by oxidation and alkylation. This protein hydrolyzes DNA adjacent to the 5'-end of an apurinic/apyrimidinic (AP) site to produce a nick with a 3'-hydroxyl group and a 5'-deoxyribose phosphate moiety or activates the DNA-binding activity of certain transcription factors through its redox function. Studies have indicated a role for APE1/Ref-1 in the pathogenesis of cancer and in resistance to DNA-interactive drugs. Thus, this protein has potential as a target in cancer treatment. As a result, major efforts have been directed to identify small molecule inhibitors against APE1/Ref-1 activities. These agents have the potential to become anticancer drugs. The aim of this review is to present recent progress in studies of all published small molecule APE1/Ref-1 inhibitors. The structures and activities of APE1/Ref-1 inhibitors, that target both DNA repair and redox activities, are presented and discussed. To date, there is an urgent need for further development of the design and synthesis of APE1/Ref-1 inhibitors due to high importance of this protein target. Copyright © 2017 Elsevier Ltd. All rights reserved.

  16. Home is Private…Do Not Enter! Introversion and Sensitivity to Work-Home Conflict.

    Science.gov (United States)

    Baer, Stacy M; Jenkins, Jade S; Barber, Larissa K

    2016-10-01

    This study examined extraversion as a moderator of the relationship between negative work-home conflict and stress-related outcomes among US employees using conservation of resources theory and privacy regulation theory. Introverts only experienced stronger negative effects of negative work-home conflict on work-related resource depletion (job burnout, low engagement, low satisfaction with balance) rather than general resource depletion (personal burnout) and strain (physical and psychological). This finding suggests that introverts selectively withdraw from the work domain to conserve resources when privacy at home is threatened. Employers may want to consider ways to help introverts increase work-home segmentation, such as reducing workplace norms that encourage employees to be continuously accessible. Copyright © 2014 John Wiley & Sons, Ltd. Copyright © 2014 John Wiley & Sons, Ltd.

  17. Prognostic value of human apurinic/apyrimidinic endonuclease 1 (APE1 expression in breast cancer.

    Directory of Open Access Journals (Sweden)

    Joohyun Woo

    Full Text Available Human apurinic/apyrimidinic endonuclease 1 (APE1 is an essential protein for DNA base excision repair (BER and redox regulation. The ability of cancer cells to recognize DNA damage and initiate DNA repair is an important mechanism for therapeutic resistance. Several recent studies have suggested that APE1 expression levels and/or subcellular dysregulation may be used to indicate the sensitivity of tumors to radiotherapy or chemotherapy. In this study, we assessed the prognostic significance of APE1 and differences in APE1 expression levels according to breast cancer molecular subtypes. We analyzed formalin-fixed, paraffin-embedded tumor tissue sections from 243 cases diagnosed as invasive breast cancer at Ewha Womans University Medical Center between January 2003 and December 2008. Immunohistochemistry was performed and the nuclear level of APE1 was scored by taking into account the percentage of positive cells. Medical records were reviewed to investigate clinicopathologic characteristics. We found that nuclear APE1 high-level expression (proportion ≥50% in breast cancer showed a tendency towards unfavorable prognosis regarding disease-free survival (p = 0.093. However, there was no significant difference in overall survival between low and high-level expression groups (p = 0.294. Interestingly, within the Ki-67 low-level expression group, APE1 low-level expression was significantly associated with poor overall survival (p = 0.007. A significant positive correlation was observed between APE1 nuclear expression and estrogen receptor status (75.7% vs. 59.7%, p = 0.022. Also, the luminal A subtype was the most commonly observed breast cancer subtype in the APE1 high-level expression group (61.6% vs. 45.2%, p = 0.000. This study suggests that APE1 expression may be associated with breast cancer prognosis. In particular, its role as a prognostic factor would be significant for breast cancers with a low Ki-67 proliferation index

  18. Apurinic/apyrimidinic endonuclease 1 regulates angiogenesis in a transforming growth factor β-dependent manner in human osteosarcoma.

    Science.gov (United States)

    Jiang, Xuan; Shan, Jinlu; Dai, Nan; Zhong, Zhaoyang; Qing, Yi; Yang, Yuxing; Zhang, Shiheng; Li, Chongyi; Sui, Jiangdong; Ren, Tao; Li, Mengxia; Wang, Dong

    2015-10-01

    Angiogenesis plays an important role in tumor growth and metastasis and has been reported to be inversely correlated with overall survival of osteosarcoma patients. It has been shown that apurinic/apyrimidinic endonuclease 1 (APE1), a dually functional protein possessing both base excision repair and redox activities, is involved in tumor angiogenesis, although these mechanisms are not fully understood. Our previous study showed that the expression of transforming growth factor β (TGFβ) was significantly reduced in APE1-deficient osteosarcoma cells. Transforming growth factor β promotes cancer metastasis through various mechanisms including immunosuppression, angiogenesis, and invasion. In the current study, we initially revealed that APE1, TGFβ, and microvessel density (MVD) have pairwise correlation in osteosarcoma tissue samples, whereas TGFβ, tumor size, and MVD were inversely related to the prognosis of the cohort. We found that knocking down APE1 in osteosarcoma cells resulted in TGFβ downregulation. In addition, APE1-siRNA led to suppression of angiogenesis in vitro based on HUVECs in Transwell and Matrigel tube formation assays. Reduced secretory protein level of TGFβ of culture medium also resulted in decreased phosphorylation of Smad3 of HUVECs. In a mouse xenograft model, siRNA-mediated silencing of APE1 downregulated TGFβ expression, tumor size, and MVD. Collectively, the current evidence indicates that APE1 regulates angiogenesis in osteosarcoma by controlling the TGFβ pathway, suggesting a novel target for anti-angiogenesis therapy in human osteosarcoma. © 2015 The Authors. Cancer Science published by Wiley Publishing Asia Pty Ltd on behalf of Japanese Cancer Association.

  19. Ostomy Home Skills Program

    Medline Plus

    Full Text Available ... kit contains: A booklet with information on the operation, home skills such as emptying and changing a pouch, problem solving, and home management. A DVD with demonstration of each skill Stoma ...

  20. Home-Use Tests - Cholesterol

    Science.gov (United States)

    ... Medical Procedures In Vitro Diagnostics Home Use Tests Cholesterol Share Tweet Linkedin Pin it More sharing options ... a home-use test kit to measure total cholesterol. What cholesterol is: Cholesterol is a fat (lipid) ...