Flexibility of centromere and kinetochore structures

PubMed Central

Burrack, Laura S.; Berman, Judith

2012-01-01

Centromeres, and the kinetochores that assemble on them, are essential for accurate chromosome segregation. Diverse centromere organization patterns and kinetochore structures have evolved in eukaryotes ranging from yeast to humans. In addition, centromere DNA and kinetochore position can vary even within individual cells. This flexibility manifests in several ways: centromere DNA sequences evolve rapidly, kinetochore positions shift in response to altered chromosome structure, and kinetochore complex numbers change in response to fluctuations in kinetochore protein levels. Despite their differences, all of these diverse structures promote efficient chromosome segregation. This robustness is inherent to chromosome segregation mechanisms and balances genome stability with adaptability. In this review, we explore the mechanisms and consequences of centromere and kinetochore flexibility as well as the benefits and limitations of different experimental model systems for studying them. PMID:22445183

Phosphoregulation of Spc105 by Mps1 and PP1 regulates Bub1 localization to kinetochores.

PubMed

London, Nitobe; Ceto, Steven; Ranish, Jeffrey A; Biggins, Sue

2012-05-22

Kinetochores are the macromolecular complexes that interact with microtubules to mediate chromosome segregation. Accurate segregation requires that kinetochores make bioriented attachments to microtubules from opposite poles. Attachments between kinetochores and microtubules are monitored by the spindle checkpoint, a surveillance system that prevents anaphase until every pair of chromosomes makes proper bioriented attachments. Checkpoint activity is correlated with the recruitment of checkpoint proteins to the kinetochore. Mps1 is a conserved protein kinase that regulates segregation and the spindle checkpoint, but few of the targets that mediate its functions have been identified. Here, we show that Mps1 is the major kinase activity that copurifies with budding yeast kinetochore particles and identify the conserved Spc105/KNL-1/blinkin kinetochore protein as a substrate. Phosphorylation of conserved MELT motifs within Spc105 recruits the Bub1 protein to kinetochores, and this is reversed by protein phosphatase I (PP1). Spc105 mutants lacking Mps1 phosphorylation sites are defective in the spindle checkpoint and exhibit growth defects. Together, these data identify Spc105 as a key target of the Mps1 kinase and show that the opposing activities of Mps1 and PP1 regulate the kinetochore localization of the Bub1 protein. Copyright © 2012 Elsevier Ltd. All rights reserved.

Conservation of the centromere/kinetochore protein ZW10.

PubMed

Starr, D A; Williams, B C; Li, Z; Etemad-Moghadam, B; Dawe, R K; Goldberg, M L

1997-09-22

Mutations in the essential Drosophila melanogaster gene zw10 disrupt chromosome segregation, producing chromosomes that lag at the metaphase plate during anaphase of mitosis and both meiotic divisions. Recent evidence suggests that the product of this gene, DmZW10, acts at the kinetochore as part of a tension-sensing checkpoint at anaphase onset. DmZW10 displays an intriguing cell cycle-dependent intracellular distribution, apparently moving from the centromere/kinetochore at prometaphase to kinetochore microtubules at metaphase, and back to the centromere/kinetochore at anaphase (Williams, B.C., M. Gatti, and M.L. Goldberg. 1996. J. Cell Biol. 134:1127-1140). We have identified ZW10-related proteins from widely diverse species with divergent centromere structures, including several Drosophilids, Caenorhabditis elegans, Arabidopsis thaliana, Mus musculus, and humans. Antibodies against the human ZW10 protein display a cell cycle-dependent staining pattern in HeLa cells strikingly similar to that previously observed for DmZW10 in dividing Drosophila cells. Injections of C. elegans ZW10 antisense RNA phenocopies important aspects of the mutant phenotype in Drosophila: these include a strong decrease in brood size, suggesting defects in meiosis or germline mitosis, a high percentage of lethality among the embryos that are produced, and the appearance of chromatin bridges at anaphase. These results indicate that at least some aspects of the functional role of the ZW10 protein in ensuring proper chromosome segregation are conserved across large evolutionary distances.

Zwint-1 is a novel Aurora B substrate required for the assembly of a dynein-binding platform on kinetochores

PubMed Central

Kasuboski, James M.; Bader, Jason R.; Vaughan, Patricia S.; Tauhata, Sinji B. F.; Winding, Michael; Morrissey, Meghan A.; Joyce, Michelle V.; Boggess, William; Vos, Larissa; Chan, Gordon K.; Hinchcliffe, Edward H.; Vaughan, Kevin T.

2011-01-01

Aurora B (AurB) is a mitotic kinase responsible for multiple aspects of mitotic progression, including assembly of the outer kinetochore. Cytoplasmic dynein is an abundant kinetochore protein whose recruitment to kinetochores requires phosphorylation. To assess whether AurB regulates recruitment of dynein to kinetochores, we inhibited AurB using ZM447439 or a kinase-dead AurB construct. Inhibition of AurB reduced accumulation of dynein at kinetochores substantially; however, this reflected a loss of dynein-associated proteins rather than a defect in dynein phosphorylation. We determined that AurB inhibition affected recruitment of the ROD, ZW10, zwilch (RZZ) complex to kinetochores but not zwint-1 or more-proximal kinetochore proteins. AurB phosphorylated zwint-1 but not ZW10 in vitro, and three novel phosphorylation sites were identified by tandem mass spectrometry analysis. Expression of a triple-Ala zwint-1 mutant blocked kinetochore assembly of RZZ-dependent proteins and induced defects in chromosome movement during prometaphase. Expression of a triple-Glu zwint-1 mutant rendered cells resistant to AurB inhibition during prometaphase. However, cells expressing the triple-Glu mutant failed to satisfy the spindle assembly checkpoint (SAC) at metaphase because poleward streaming of dynein/dynactin/RZZ was inhibited. These studies identify zwint-1 as a novel AurB substrate required for kinetochore assembly and for proper SAC silencing at metaphase. PMID:21775627

Evolutionary turnover of kinetochore proteins: a ship of Theseus?

PubMed Central

Drinnenberg, Ines A.; Henikoff, Steven; Malik, Harmit S.

2016-01-01

Summary The kinetochore is a multi-protein complex that mediates the attachment of a eukaryotic chromosome to the mitotic spindle. The protein composition of kinetochores is similar across species as divergent as yeast and human. However, recent findings have revealed an unexpected degree of compositional diversity in kinetochores. For example, kinetochore proteins that are essential in some species have been lost in others, whereas new kinetochore proteins have emerged in other lineages. Even in lineages with similar kinetochore composition, individual kinetochore proteins have functionally diverged to acquire either essential or redundant roles. Thus, despite functional conservation, the repertoire of kinetochore proteins has undergone recurrent evolutionary turnover. PMID:26877204

Chromosome Segregation Is Biased by Kinetochore Size.

PubMed

Drpic, Danica; Almeida, Ana C; Aguiar, Paulo; Renda, Fioranna; Damas, Joana; Lewin, Harris A; Larkin, Denis M; Khodjakov, Alexey; Maiato, Helder

2018-05-07

Chromosome missegregation during mitosis or meiosis is a hallmark of cancer and the main cause of prenatal death in humans. The gain or loss of specific chromosomes is thought to be random, with cell viability being essentially determined by selection. Several established pathways including centrosome amplification, sister-chromatid cohesion defects, or a compromised spindle assembly checkpoint can lead to chromosome missegregation. However, how specific intrinsic features of the kinetochore-the critical chromosomal interface with spindle microtubules-impact chromosome segregation remains poorly understood. Here we used the unique cytological attributes of female Indian muntjac, the mammal with the lowest known chromosome number (2n = 6), to characterize and track individual chromosomes with distinct kinetochore size throughout mitosis. We show that centromere and kinetochore functional layers scale proportionally with centromere size. Measurement of intra-kinetochore distances, serial-section electron microscopy, and RNAi against key kinetochore proteins confirmed a standard structural and functional organization of the Indian muntjac kinetochores and revealed that microtubule binding capacity scales with kinetochore size. Surprisingly, we found that chromosome segregation in this species is not random. Chromosomes with larger kinetochores bi-oriented more efficiently and showed a 2-fold bias to congress to the equator in a motor-independent manner. Despite robust correction mechanisms during unperturbed mitosis, chromosomes with larger kinetochores were also strongly biased to establish erroneous merotelic attachments and missegregate during anaphase. This bias was impervious to the experimental attenuation of polar ejection forces on chromosome arms by RNAi against the chromokinesin Kif4a. Thus, kinetochore size is an important determinant of chromosome segregation fidelity. Copyright © 2018 The Author(s). Published by Elsevier Ltd.. All rights reserved.

Evolutionary Turnover of Kinetochore Proteins: A Ship of Theseus?

PubMed

Drinnenberg, Ines A; Henikoff, Steven; Malik, Harmit S

2016-07-01

The kinetochore is a multiprotein complex that mediates the attachment of a eukaryotic chromosome to the mitotic spindle. The protein composition of kinetochores is similar across species as divergent as yeast and human. However, recent findings have revealed an unexpected degree of compositional diversity in kinetochores. For example, kinetochore proteins that are essential in some species have been lost in others, whereas new kinetochore proteins have emerged in other lineages. Even in lineages with similar kinetochore composition, individual kinetochore proteins have functionally diverged to acquire either essential or redundant roles. Thus, despite functional conservation, the repertoire of kinetochore proteins has undergone recurrent evolutionary turnover. Copyright © 2016 Elsevier Ltd. All rights reserved.

Stability of kinetochore-microtubule attachment and the role of different KMN network components in Drosophila.

PubMed

Feijão, Tália; Afonso, Olga; Maia, André F; Sunkel, Claudio E

2013-10-01

Kinetochores bind spindle microtubules and also act as signaling centers that monitor this interaction. Defects in kinetochore assembly lead to chromosome missegregation and aneuploidy. The interaction between microtubules and chromosomes involves a conserved super-complex of proteins, known as the KNL1Mis12Ndc80 (KMN) network, composed by the KNL1 (Spc105), Mis12, and Ndc80 complexes. Previous studies indicate that all components of the network are required for kinetochore-microtubule attachment and all play relevant functions in chromosome congression, biorientation, and segregation. Here, we report a comparative study addressing the role of the different KMN components using dsRNA and in vivo fluorescence microscopy in Drosophila S2 cells allowing us to suggest that different KMN network components might perform different roles in chromosome segregation and the mitotic checkpoint signaling. Depletion of different components results in mostly lateral kinetochore-microtubule attachments that are relatively stable on depletion of Mis12 or Ndc80 but very unstable after Spc105 depletion. In vivo analysis on depletion of Mis12, Ndc80, and to some extent Spc105, shows that lateral kinetochore-microtubule interactions are still functional allowing poleward kinetochore movement. We also find that different KMN network components affect differently the localization of spindle assembly checkpoint (SAC) proteins at kinetochores. Depletion of Ndc80 and Spc105 abolishes the mitotic checkpoint, whereas depletion of Mis12 causes a delay in mitotic progression. Taken together, our results suggest that Mis12 and Ndc80 complexes help to properly orient microtubule attachment, whereas Spc105 plays a predominant role in the kinetochore-microtubule attachment as well as in the poleward movement of chromosomes, SAC response, and cell viability. Copyright © 2013 Wiley Periodicals, Inc.

The life and miracles of kinetochores

PubMed Central

Santaguida, Stefano; Musacchio, Andrea

2009-01-01

Kinetochores are large protein assemblies built on chromosomal loci named centromeres. The main functions of kinetochores can be grouped under four modules. The first module, in the inner kinetochore, contributes a sturdy interface with centromeric chromatin. The second module, the outer kinetochore, contributes a microtubule-binding interface. The third module, the spindle assembly checkpoint, is a feedback control mechanism that monitors the state of kinetochore–microtubule attachment to control the progression of the cell cycle. The fourth module discerns correct from improper attachments, preventing the stabilization of the latter and allowing the selective stabilization of the former. In this review, we discuss how the molecular organization of the four modules allows a dynamic integration of kinetochore–microtubule attachment with the prevention of chromosome segregation errors and cell-cycle progression. PMID:19629042

Molecular mechanisms of microtubule-dependent kinetochore transport toward spindle poles

PubMed Central

Tanaka, Kozo; Kitamura, Etsushi; Kitamura, Yoko; Tanaka, Tomoyuki U.

2007-01-01

In mitosis, kinetochores are initially captured by the lateral sides of single microtubules and are subsequently transported toward spindle poles. Mechanisms for kinetochore transport are not yet known. We present two mechanisms involved in microtubule-dependent poleward kinetochore transport in Saccharomyces cerevisiae. First, kinetochores slide along the microtubule lateral surface, which is mainly and probably exclusively driven by Kar3, a kinesin-14 family member that localizes at kinetochores. Second, kinetochores are tethered at the microtubule distal ends and pulled poleward as microtubules shrink (end-on pulling). Kinetochore sliding is often converted to end-on pulling, enabling more processive transport, but the opposite conversion is rare. The establishment of end-on pulling is partly hindered by Kar3, and its progression requires the Dam1 complex. We suggest that the Dam1 complexes, which probably encircle a single microtubule, can convert microtubule depolymerization into the poleward kinetochore-pulling force. Thus, microtubule-dependent poleward kinetochore transport is ensured by at least two distinct mechanisms. PMID:17620411

Immunofluorescence Analysis of Endogenous and Exogenous Centromere-kinetochore Proteins

PubMed Central

Niikura, Yohei; Kitagawa, Katsumi

2016-01-01

"Centromeres" and "kinetochores" refer to the site where chromosomes associate with the spindle during cell division. Direct visualization of centromere-kinetochore proteins during the cell cycle remains a fundamental tool in investigating the mechanism(s) of these proteins. Advanced imaging methods in fluorescence microscopy provide remarkable resolution of centromere-kinetochore components and allow direct observation of specific molecular components of the centromeres and kinetochores. In addition, methods of indirect immunofluorescent (IIF) staining using specific antibodies are crucial to these observations. However, despite numerous reports about IIF protocols, few discussed in detail problems of specific centromere-kinetochore proteins.1-4 Here we report optimized protocols to stain endogenous centromere-kinetochore proteins in human cells by using paraformaldehyde fixation and IIF staining. Furthermore, we report protocols to detect Flag-tagged exogenous CENP-A proteins in human cells subjected to acetone or methanol fixation. These methods are useful in detecting and quantifying endogenous centromere-kinetochore proteins and Flag-tagged CENP-A proteins, including those in human cells. PMID:26967065

Basic mechanism for biorientation of mitotic chromosomes is provided by the kinetochore geometry and indiscriminate turnover of kinetochore microtubules

PubMed Central

Zaytsev, Anatoly V.; Grishchuk, Ekaterina L.

2015-01-01

Accuracy of chromosome segregation relies on the ill-understood ability of mitotic kinetochores to biorient, whereupon each sister kinetochore forms microtubule (MT) attachments to only one spindle pole. Because initial MT attachments result from chance encounters with the kinetochores, biorientation must rely on specific mechanisms to avoid and resolve improper attachments. Here we use mathematical modeling to critically analyze the error-correction potential of a simplified biorientation mechanism, which involves the back-to-back arrangement of sister kinetochores and the marked instability of kinetochore–MT attachments. We show that a typical mammalian kinetochore operates in a near-optimal regime, in which the back-to-back kinetochore geometry and the indiscriminate kinetochore–MT turnover provide strong error-correction activity. In human cells, this mechanism alone can potentially enable normal segregation of 45 out of 46 chromosomes during one mitotic division, corresponding to a mis-segregation rate in the range of 10−1–10−2 per chromosome. This theoretical upper limit for chromosome segregation accuracy predicted with the basic mechanism is close to the mis-segregation rate in some cancer cells; however, it cannot explain the relatively low chromosome loss in diploid human cells, consistent with their reliance on additional mechanisms. PMID:26424798

A Molecular View of Kinetochore Assembly and Function

PubMed Central

Musacchio, Andrea; Desai, Arshad

2017-01-01

Kinetochores are large protein assemblies that connect chromosomes to microtubules of the mitotic and meiotic spindles in order to distribute the replicated genome from a mother cell to its daughters. Kinetochores also control feedback mechanisms responsible for the correction of incorrect microtubule attachments, and for the coordination of chromosome attachment with cell cycle progression. Finally, kinetochores contribute to their own preservation, across generations, at the specific chromosomal loci devoted to host them, the centromeres. They achieve this in most species by exploiting an epigenetic, DNA-sequence-independent mechanism; notable exceptions are budding yeasts where a specific sequence is associated with centromere function. In the last 15 years, extensive progress in the elucidation of the composition of the kinetochore and the identification of various physical and functional modules within its substructure has led to a much deeper molecular understanding of kinetochore organization and the origins of its functional output. Here, we provide a broad summary of this progress, focusing primarily on kinetochores of humans and budding yeast, while highlighting work from other models, and present important unresolved questions for future studies. PMID:28125021

Pac-man motility of kinetochores unleashed by laser microsurgery

PubMed Central

LaFountain, James R.; Cohan, Christopher S.; Oldenbourg, Rudolf

2012-01-01

We report on experiments directly in living cells that reveal the regulation of kinetochore function by tension. X and Y sex chromosomes in crane fly (Nephrotoma suturalis) spermatocytes exhibit an atypical segregation mechanism in which each univalent maintains K-fibers to both poles. During anaphase, each maintains a leading fiber (which shortens) to one pole and a trailing fiber (which elongates) to the other. We used this intriguing behavior to study the motile states that X-Y kinetochores are able to support during anaphase. We used a laser microbeam to either sever a univalent along the plane of sister chromatid cohesion or knock out one of a univalent's two kinetochores to release one or both from the resistive influence of its sister's K-fiber. Released kinetochores with attached chromosome arms moved poleward at rates at least two times faster than normal. Furthermore, fluorescent speckle microscopy revealed that detached kinetochores converted their functional state from reverse pac-man to pac-man motility as a consequence of their release from mechanical tension. We conclude that kinetochores can exhibit pac-man motility, even though their normal behavior is dominated by traction fiber mechanics. Unleashing of kinetochore motility through loss of resistive force is further evidence for the emerging model that kinetochores are subject to tension-sensitive regulation. PMID:22740625

Evolving Centromeres and Kinetochores.

PubMed

Friedman, Steven; Freitag, Michael

2017-01-01

The genetic material, contained on chromosomes, is often described as the "blueprint for life." During nuclear division, the chromosomes are pulled into each of the two daughter nuclei by the coordination of spindle microtubules, kinetochores, centromeres, and chromatin. These four functional units must link the chromosomes to the microtubules, signal to the cell when the attachment is made so that division can proceed, and withstand the force generated by pulling the chromosomes to either daughter cell. To perform each of these functions, kinetochores are large protein complexes, approximately 5MDa in size, and they contain at least 45 unique proteins. Many of the central components in the kinetochore are well conserved, yielding a common core of proteins forming consistent structures. However, many of the peripheral subcomplexes vary between different taxonomic groups, including changes in primary sequence and gain or loss of whole proteins. It is still unclear how significant these changes are, and answers to this question may provide insights into adaptation to specific lifestyles or progression of disease that involve chromosome instability. Copyright © 2017 Elsevier Inc. All rights reserved.

Modular elements of the TPR domain in the Mps1 N terminus differentially target Mps1 to the centrosome and kinetochore

PubMed Central

Marquardt, Joseph R.; Perkins, Jennifer L.; Beuoy, Kyle J.; Fisk, Harold A.

2016-01-01

Faithful segregation of chromosomes to two daughter cells is regulated by the formation of a bipolar mitotic spindle and the spindle assembly checkpoint, ensuring proper spindle function. Here we show that the proper localization of the kinase Mps1 (monopolar spindle 1) is critical to both these processes. Separate elements in the Mps1 N-terminal extension (NTE) and tetratricopeptide repeat (TPR) domains govern localization to either the kinetochore or the centrosome. The third TPR (TPR3) and the TPR-capping helix (C-helix) are each sufficient to target Mps1 to the centrosome. TPR3 binds to voltage-dependent anion channel 3, but although this is sufficient for centrosome targeting of Mps1, it is not necessary because of the presence of the C-helix. A version of Mps1 lacking both elements cannot localize to or function at the centrosome, but maintains kinetochore localization and spindle assembly checkpoint function, indicating that TPR3 and the C-helix define a bipartite localization determinant that is both necessary and sufficient to target Mps1 to the centrosome but dispensable for kinetochore targeting. In contrast, elements required for kinetochore targeting (the NTE and first two TPRs) are dispensable for centrosomal localization and function. These data are consistent with a separation of Mps1 function based on localization determinants within the N terminus. PMID:27339139

Modular elements of the TPR domain in the Mps1 N terminus differentially target Mps1 to the centrosome and kinetochore.

PubMed

Marquardt, Joseph R; Perkins, Jennifer L; Beuoy, Kyle J; Fisk, Harold A

2016-07-12

Faithful segregation of chromosomes to two daughter cells is regulated by the formation of a bipolar mitotic spindle and the spindle assembly checkpoint, ensuring proper spindle function. Here we show that the proper localization of the kinase Mps1 (monopolar spindle 1) is critical to both these processes. Separate elements in the Mps1 N-terminal extension (NTE) and tetratricopeptide repeat (TPR) domains govern localization to either the kinetochore or the centrosome. The third TPR (TPR3) and the TPR-capping helix (C-helix) are each sufficient to target Mps1 to the centrosome. TPR3 binds to voltage-dependent anion channel 3, but although this is sufficient for centrosome targeting of Mps1, it is not necessary because of the presence of the C-helix. A version of Mps1 lacking both elements cannot localize to or function at the centrosome, but maintains kinetochore localization and spindle assembly checkpoint function, indicating that TPR3 and the C-helix define a bipartite localization determinant that is both necessary and sufficient to target Mps1 to the centrosome but dispensable for kinetochore targeting. In contrast, elements required for kinetochore targeting (the NTE and first two TPRs) are dispensable for centrosomal localization and function. These data are consistent with a separation of Mps1 function based on localization determinants within the N terminus.

Complex regulation of sister kinetochore orientation in meiosis-I.

PubMed

Bardhan, Amit

2010-09-01

Kinetochores mediate chromosome movement during cell division by interacting with the spindle microtubules. Sexual reproduction necessitates the daunting task of reducing ploidy (number of chromosome sets) in the gametes, which depends upon the specialized properties of meiosis. Kinetochores have a central role in the reduction process. In this review, we discuss the complexity of this role of kinetochores in meiosis-I.

Sister kinetochores are mechanically fused during meiosis I in yeast.

PubMed

Sarangapani, Krishna K; Duro, Eris; Deng, Yi; Alves, Flavia de Lima; Ye, Qiaozhen; Opoku, Kwaku N; Ceto, Steven; Rappsilber, Juri; Corbett, Kevin D; Biggins, Sue; Marston, Adèle L; Asbury, Charles L

2014-10-10

Production of healthy gametes requires a reductional meiosis I division in which replicated sister chromatids comigrate, rather than separate as in mitosis or meiosis II. Fusion of sister kinetochores during meiosis I may underlie sister chromatid comigration in diverse organisms, but direct evidence for such fusion has been lacking. We used laser trapping and quantitative fluorescence microscopy to study native kinetochore particles isolated from yeast. Meiosis I kinetochores formed stronger attachments and carried more microtubule-binding elements than kinetochores isolated from cells in mitosis or meiosis II. The meiosis I-specific monopolin complex was both necessary and sufficient to drive these modifications. Thus, kinetochore fusion directs sister chromatid comigration, a conserved feature of meiosis that is fundamental to Mendelian inheritance. Copyright © 2014, American Association for the Advancement of Science.

Centromeres and kinetochores of Brassicaceae.

PubMed

Lermontova, Inna; Sandmann, Michael; Demidov, Dmitri

2014-06-01

The centromere-the primary constriction of monocentric chromosomes-is essential for correct segregation of chromosomes during mitosis and meiosis. Centromeric DNA varies between different organisms in sequence composition and extension. The main components of centromeric and pericentromeric DNA of Brassicaceae species are centromeric satellite repeats. Centromeric DNA initiates assembly of the kinetochore, the large protein complex where the spindle fibers attach during nuclear division to pull sister chromatids apart. Kinetochore assembly is initiated by incorporation of the centromeric histone H3 cenH3 into centromeric nucleosomes. The spindle assembly checkpoint acts during mitosis and meiosis at centromeres and maintains genome stability by preventing chromosome segregation before all kinetochores are correctly attached to microtubules. The function of the spindle assembly checkpoint in plants is still poorly understood. Here, we review recent advances of studies on structure and functional importance of centromeric DNA of Brassicaceae, assembly and function of cenH3 in Arabidopsis thaliana and characterization of core SAC proteins of A. thaliana in comparison with non-plant homologues.

Regulation of outer kinetochore Ndc80 complex-based microtubule attachments by the central kinetochore Mis12/MIND complex

PubMed Central

Kudalkar, Emily M.; Scarborough, Emily A.; Umbreit, Neil T.; Zelter, Alex; Gestaut, Daniel R.; Riffle, Michael; Johnson, Richard S.; MacCoss, Michael J.; Asbury, Charles L.; Davis, Trisha N.

2015-01-01

Multiple protein subcomplexes of the kinetochore cooperate as a cohesive molecular unit that forms load-bearing microtubule attachments that drive mitotic chromosome movements. There is intriguing evidence suggesting that central kinetochore components influence kinetochore–microtubule attachment, but the mechanism remains unclear. Here, we find that the conserved Mis12/MIND (Mtw1, Nsl1, Nnf1, Dsn1) and Ndc80 (Ndc80, Nuf2, Spc24, Spc25) complexes are connected by an extensive network of contacts, each essential for viability in cells, and collectively able to withstand substantial tensile load. Using a single-molecule approach, we demonstrate that an individual MIND complex enhances the microtubule-binding affinity of a single Ndc80 complex by fourfold. MIND itself does not bind microtubules. Instead, MIND binds Ndc80 complex far from the microtubule-binding domain and confers increased microtubule interaction of the complex. In addition, MIND activation is redundant with the effects of a mutation in Ndc80 that might alter its ability to adopt a folded conformation. Together, our results suggest a previously unidentified mechanism for regulating microtubule binding of an outer kinetochore component by a central kinetochore complex. PMID:26430240

Kinetochores: if you build it, they will come.

PubMed

DeLuca, Jennifer G; Salmon, E D

2004-11-09

Successful mitosis depends critically on the segregation of chromosomes by kinetochore microtubules. A recent paper describes a conserved protein network from Caenorhabditis elegans that is composed of three classes of molecules, each of which contributes uniquely to the building of the kinetochore-microtubule attachment site.

KiT: a MATLAB package for kinetochore tracking.

PubMed

Armond, Jonathan W; Vladimirou, Elina; McAinsh, Andrew D; Burroughs, Nigel J

2016-06-15

During mitosis, chromosomes are attached to the mitotic spindle via large protein complexes called kinetochores. The motion of kinetochores throughout mitosis is intricate and automated quantitative tracking of their motion has already revealed many surprising facets of their behaviour. Here, we present 'KiT' (Kinetochore Tracking)-an easy-to-use, open-source software package for tracking kinetochores from live-cell fluorescent movies. KiT supports 2D, 3D and multi-colour movies, quantification of fluorescence, integrated deconvolution, parallel execution and multiple algorithms for particle localization. KiT is free, open-source software implemented in MATLAB and runs on all MATLAB supported platforms. KiT can be downloaded as a package from http://www.mechanochemistry.org/mcainsh/software.php The source repository is available at https://bitbucket.org/jarmond/kit and under continuing development. Supplementary data are available at Bioinformatics online. jonathan.armond@warwick.ac.uk. © The Author 2016. Published by Oxford University Press.

Congressing kinetochores progressively load Ska complexes to prevent force-dependent detachment

PubMed Central

Auckland, Philip; Clarke, Nicholas I.

2017-01-01

Kinetochores mediate chromosome congression by either sliding along the lattice of spindle microtubules or forming end-on attachments to their depolymerizing plus-ends. By following the fates of individual kinetochores as they congress in live cells, we reveal that the Ska complex is required for a distinct substep of the depolymerization-coupled pulling mechanism. Ska depletion increases the frequency of naturally occurring, force-dependent P kinetochore detachment events, while being dispensable for the initial biorientation and movement of chromosomes. In unperturbed cells, these release events are followed by reattachment and successful congression, whereas in Ska-depleted cells, detached kinetochores remain in a futile reattachment/detachment cycle that prevents congression. We further find that Ska is progressively loaded onto bioriented kinetochore pairs as they congress. We thus propose a model in which kinetochores mature through Ska complex recruitment and that this is required for improved load-bearing capacity and silencing of the spindle assembly checkpoint. PMID:28495837

Congressing kinetochores progressively load Ska complexes to prevent force-dependent detachment.

PubMed

Auckland, Philip; Clarke, Nicholas I; Royle, Stephen J; McAinsh, Andrew D

2017-06-05

Kinetochores mediate chromosome congression by either sliding along the lattice of spindle microtubules or forming end-on attachments to their depolymerizing plus-ends. By following the fates of individual kinetochores as they congress in live cells, we reveal that the Ska complex is required for a distinct substep of the depolymerization-coupled pulling mechanism. Ska depletion increases the frequency of naturally occurring, force-dependent P kinetochore detachment events, while being dispensable for the initial biorientation and movement of chromosomes. In unperturbed cells, these release events are followed by reattachment and successful congression, whereas in Ska-depleted cells, detached kinetochores remain in a futile reattachment/detachment cycle that prevents congression. We further find that Ska is progressively loaded onto bioriented kinetochore pairs as they congress. We thus propose a model in which kinetochores mature through Ska complex recruitment and that this is required for improved load-bearing capacity and silencing of the spindle assembly checkpoint. © 2017 Auckland et al.

Joined at the hip: kinetochores, microtubules, and spindle assembly checkpoint signaling.

PubMed

Sacristan, Carlos; Kops, Geert J P L

2015-01-01

Error-free chromosome segregation relies on stable connections between kinetochores and spindle microtubules. The spindle assembly checkpoint (SAC) monitors such connections and relays their absence to the cell cycle machinery to delay cell division. The molecular network at kinetochores that is responsible for microtubule binding is integrated with the core components of the SAC signaling system. Molecular-mechanistic understanding of how the SAC is coupled to the kinetochore-microtubule interface has advanced significantly in recent years. The latest insights not only provide a striking view of the dynamics and regulation of SAC signaling events at the outer kinetochore but also create a framework for understanding how that signaling may be terminated when kinetochores and microtubules connect. Copyright © 2014 Elsevier Ltd. All rights reserved.

The Ndc80 complex targets Bod1 to human mitotic kinetochores

PubMed Central

2017-01-01

Regulation of protein phosphatase activity by endogenous protein inhibitors is an important mechanism to control protein phosphorylation in cells. We recently identified Biorientation defective 1 (Bod1) as a small protein inhibitor of protein phosphatase 2A containing the B56 regulatory subunit (PP2A-B56). This phosphatase controls the amount of phosphorylation of several kinetochore proteins and thus the establishment of load-bearing chromosome-spindle attachments in time for accurate separation of sister chromatids in mitosis. Like PP2A-B56, Bod1 directly localizes to mitotic kinetochores and is required for correct segregation of mitotic chromosomes. In this report, we have probed the spatio-temporal regulation of Bod1 during mitotic progression. Kinetochore localization of Bod1 increases from nuclear envelope breakdown until metaphase. Phosphorylation of Bod1 at threonine 95 (T95), which increases Bod1's binding to and inhibition of PP2A-B56, peaks in prometaphase when PP2A-B56 localization to kinetochores is highest. We demonstrate here that kinetochore targeting of Bod1 depends on the outer kinetochore protein Ndc80 and not PP2A-B56. Crucially, Bod1 depletion functionally affects Ndc80 phosphorylation at the N-terminal serine 55 (S55), as well as a number of other phosphorylation sites within the outer kinetochore, including Knl1 at serine 24 and 60 (S24, S60), and threonine T943 and T1155 (T943, T1155). Therefore, Ndc80 recruits a phosphatase inhibitor to kinetochores which directly feeds forward to regulate Ndc80, and Knl1 phosphorylation, including sites that mediate the attachment of microtubules to kinetochores. PMID:29142109

The Centromere: Chromatin Foundation for the Kinetochore Machinery

PubMed Central

Fukagawa, Tatsuo; Earnshaw, William C.

2014-01-01

Since discovery of the centromere-specific histone H3 variant CENP-A, centromeres have come to be defined as chromatin structures that establish the assembly site for the complex kinetochore machinery. In most organisms, centromere activity is defined epigenetically, rather than by specific DNA sequences. In this review, we describe selected classic work and recent progress in studies of centromeric chromatin with a focus on vertebrates. We consider possible roles for repetitive DNA sequences found at most centromeres, chromatin factors and modifications that assemble and activate CENP-A chromatin for kinetochore assembly, plus the use of artificial chromosomes and kinetochores to study centromere function. PMID:25203206

Architecture and Flexibility of the Yeast Ndc80 Kinetochore Complex

PubMed Central

Wang, Hong-Wei; Long, Sydney; Ciferri, Claudio; Westermann, Stefan; Drubin, David; Barnes, Georjana; Nogales, Eva

2008-01-01

Kinetochores mediate microtubule–chromosome attachment and ensure accurate segregation of sister chromatids. The highly conserved Ndc80 kinetochore complex makes direct contacts with the microtubule and is essential for spindle checkpoint signaling. It contains a long coiled-coil region with globular domains at each end involved in kinetochore localization and microtubule binding, respectively. We have directly visualized the architecture of the yeast Ndc80 complex and found a dramatic kink within the 560-Å coiled-coil rod located about 160 Å from the larger globular head. Comparison of our electron microscopy images to the structure of the human Ndc80 complex allowed us to position the kink proximal to the microtubule-binding end and to define the conformational range of the complex. The position of the kink coincides with a coiled-coil breaking region conserved across eukaryotes. We hypothesize that the kink in Ndc80 is essential for correct kinetochore geometry and could be part of a tension-sensing mechanism at the kinetochore. PMID:18793650

Overlap microtubules link sister k-fibres and balance the forces on bi-oriented kinetochores

PubMed Central

Kajtez, Janko; Solomatina, Anastasia; Novak, Maja; Polak, Bruno; Vukušić, Kruno; Rüdiger, Jonas; Cojoc, Gheorghe; Milas, Ana; Šumanovac Šestak, Ivana; Risteski, Patrik; Tavano, Federica; Klemm, Anna H.; Roscioli, Emanuele; Welburn, Julie; Cimini, Daniela; Glunčić, Matko; Pavin, Nenad; Tolić, Iva M.

2016-01-01

During metaphase, forces on kinetochores are exerted by k-fibres, bundles of microtubules that end at the kinetochore. Interestingly, non-kinetochore microtubules have been observed between sister kinetochores, but their function is unknown. Here we show by laser-cutting of a k-fibre in HeLa and PtK1 cells that a bundle of non-kinetochore microtubules, which we term ‘bridging fibre', bridges sister k-fibres and balances the interkinetochore tension. We found PRC1 and EB3 in the bridging fibre, suggesting that it consists of antiparallel dynamic microtubules. By using a theoretical model that includes a bridging fibre, we show that the forces at the pole and at the kinetochore depend on the bridging fibre thickness. Moreover, our theory and experiments show larger relaxation of the interkinetochore distance for cuts closer to kinetochores. We conclude that the bridging fibre, by linking sister k-fibres, withstands the tension between sister kinetochores and enables the spindle to obtain a curved shape. PMID:26728792

Meiosis I chromosome segregation is established through regulation of microtubule–kinetochore interactions

PubMed Central

Miller, Matthew P; Ünal, Elçin; Brar, Gloria A; Amon, Angelika

2012-01-01

During meiosis, a single round of DNA replication is followed by two consecutive rounds of nuclear divisions called meiosis I and meiosis II. In meiosis I, homologous chromosomes segregate, while sister chromatids remain together. Determining how this unusual chromosome segregation behavior is established is central to understanding germ cell development. Here we show that preventing microtubule–kinetochore interactions during premeiotic S phase and prophase I is essential for establishing the meiosis I chromosome segregation pattern. Premature interactions of kinetochores with microtubules transform meiosis I into a mitosis-like division by disrupting two key meiosis I events: coorientation of sister kinetochores and protection of centromeric cohesin removal from chromosomes. Furthermore we find that restricting outer kinetochore assembly contributes to preventing premature engagement of microtubules with kinetochores. We propose that inhibition of microtubule–kinetochore interactions during premeiotic S phase and prophase I is central to establishing the unique meiosis I chromosome segregation pattern. DOI: http://dx.doi.org/10.7554/eLife.00117.001 PMID:23275833

Dynamics and control of sister kinetochore behavior during the meiotic divisions in Drosophila spermatocytes

PubMed Central

2018-01-01

Sister kinetochores are connected to the same spindle pole during meiosis I and to opposite poles during meiosis II. The molecular mechanisms controlling the distinct behavior of sister kinetochores during the two meiotic divisions are poorly understood. To study kinetochore behavior during meiosis, we have optimized time lapse imaging with Drosophila spermatocytes, enabling kinetochore tracking with high temporal and spatial resolution through both meiotic divisions. The correct bipolar orientation of chromosomes within the spindle proceeds rapidly during both divisions. Stable bi-orientation of the last chromosome is achieved within ten minutes after the onset of kinetochore-microtubule interactions. Our analyses of mnm and tef mutants, where univalents instead of bivalents are present during meiosis I, indicate that the high efficiency of normal bi-orientation depends on pronounced stabilization of kinetochore attachments to spindle microtubules by the mechanical tension generated by spindle forces upon bi-orientation. Except for occasional brief separation episodes, sister kinetochores are so closely associated that they cannot be resolved individually by light microscopy during meiosis I, interkinesis and at the start of meiosis II. Permanent evident separation of sister kinetochores during M II depends on spindle forces resulting from bi-orientation. In mnm and tef mutants, sister kinetochore separation can be observed already during meiosis I in bi-oriented univalents. Interestingly, however, this sister kinetochore separation is delayed until the metaphase to anaphase transition and depends on the Fzy/Cdc20 activator of the anaphase-promoting complex/cyclosome. We propose that univalent bi-orientation in mnm and tef mutants exposes a release of sister kinetochore conjunction that occurs also during normal meiosis I in preparation for bi-orientation of dyads during meiosis II. PMID:29734336

Kinetochore Dynein Is Required for Chromosome Motion and Congression Independent of the Spindle Checkpoint

PubMed Central

Yang, Zhenye; Tulu, U. Serdar; Wadsworth, Patricia; Rieder, Conly L.

2008-01-01

Summary During mitosis, the motor molecule cytoplasmic dynein plays key direct and indirect roles in organizing microtubules (MTs) into a functional spindle. At this time, dynein is also recruited to kinetochores, but its role or roles at these organelles remain vague, partly because inhibiting dynein globally disrupts spindle assembly [1-4]. However, dynein can be selectively depleted from kinetochores by disruption of ZW10 [5], and recent studies with this approach conclude that kinetochore-associated dynein (KD) functions to silence the spindle-assembly checkpoint (SAC) [6]. Here we use dynein-antibody microinjection and the RNAi of ZW10 to explore the role of KD in chromosome behavior during mitosis in mammals. We find that depleting or inhibiting KD prevents the rapid poleward motion of attaching kinetochores but not kinetochore fiber (K fiber) formation. However, after kinetochores attach to the spindle, KD is required for stabilizing kinetochore MTs, which it probably does by generating tension on the kinetochore, and in its absence, chromosome congression is defective. Finally, depleting KD reduces the velocity of anaphase chromosome motion by ∼40%, without affecting the rate of poleward MT flux. Thus, in addition to its role in silencing the SAC, KD is important for forming and stabilizing K fibers and in powering chromosome motion. PMID:17509882

BuGZ is required for Bub3 stability, Bub1 kinetochore function, and chromosome alignment

PubMed Central

Toledo, Chad M.; Herman, Jacob A.; Olsen, Jonathan B.; Ding, Yu; Corrin, Philip; Girard, Emily J.; Olson, James M.; Emili, Andrew; DeLuca, Jennifer G.; Paddison, Patrick J.

2014-01-01

Summary During mitosis, the spindle assembly checkpoint (SAC) monitors the attachment of kinetochores (KTs) to the plus ends of spindle microtubules (MTs) and prevents anaphase onset until chromosomes are aligned and KTs are under proper tension. Here, we identify a SAC component, BuGZ/ZNF207, from an RNAi viability screen in human Glioblastoma multiforme (GBM) brain tumor stem cells. BuGZ binds to and stabilizes Bub3 during interphase and mitosis through a highly conserved GLE2p-binding sequence (GLEBS) domain. Inhibition of BuGZ results in loss of both Bub3 and its binding partner Bub1 from KTs, reduction of Bub1-dependent phosphorylation of centromeric histone H2A, attenuation of KT-based Aurora kinase B activity, and lethal chromosome congression defects in cancer cells. Phylogenetic analysis indicates that BuGZ orthologs are highly conserved among eukaryotes, but are conspicuously absent from budding and fission yeasts. These findings suggest BuGZ has evolved to facilitate Bub3 activity and chromosome congression in higher eukaryotes. PMID:24462187

RED, a Spindle Pole-associated Protein, Is Required for Kinetochore Localization of MAD1, Mitotic Progression, and Activation of the Spindle Assembly Checkpoint*

PubMed Central

Yeh, Pei-Chi; Yeh, Chang-Ching; Chen, Yi-Cheng; Juang, Yue-Li

2012-01-01

The spindle assembly checkpoint (SAC) is essential for ensuring the proper attachment of kinetochores to the spindle and, thus, the precise separation of paired sister chromatids during mitosis. The SAC proteins are recruited to the unattached kinetochores for activation of the SAC in prometaphase. However, it has been less studied whether activation of the SAC also requires the proteins that do not localize to the kinetochores. Here, we show that the nuclear protein RED, also called IK, a down-regulator of human leukocyte antigen (HLA) II, interacts with the human SAC protein MAD1. Two RED-interacting regions identified in MAD1 are from amino acid residues 301–340 and 439–480, designated as MAD1(301–340) and MAD1(439–480), respectively. Our observations reveal that RED is a spindle pole-associated protein that colocalizes with MAD1 at the spindle poles in metaphase and anaphase. Depletion of RED can cause a shorter mitotic timing, a failure in the kinetochore localization of MAD1 in prometaphase, and a defect in the SAC. Furthermore, the RED-interacting peptides MAD1(301–340) and MAD1(439–480), fused to enhanced green fluorescence protein, can colocalize with RED at the spindle poles in prometaphase, and their expression can abrogate the SAC. Taken together, we conclude that RED is required for kinetochore localization of MAD1, mitotic progression, and activation of the SAC. PMID:22351768

Synthetic physical interactions map kinetochore regulators and regions sensitive to constitutive Cdc14 localization.

PubMed

Ólafsson, Guðjón; Thorpe, Peter H

2015-08-18

The location of proteins within eukaryotic cells is often critical for their function and relocation of proteins forms the mainstay of regulatory pathways. To assess the importance of protein location to cellular homeostasis, we have developed a methodology to systematically create binary physical interactions between a query protein and most other members of the proteome. This method allows us to rapidly assess which of the thousands of possible protein interactions modify a phenotype. As proof of principle we studied the kinetochore, a multiprotein assembly that links centromeres to the microtubules of the spindle during cell division. In budding yeast, the kinetochores from the 16 chromosomes cluster together to a single location within the nucleus. The many proteins that make up the kinetochore are regulated through ubiquitylation and phosphorylation. By systematically associating members of the proteome to the kinetochore, we determine which fusions affect its normal function. We identify a number of candidate kinetochore regulators, including the phosphatase Cdc14. We examine where within the kinetochore Cdc14 can act and show that the effect is limited to regions that correlate with known phosphorylation sites, demonstrating the importance of serine phospho-regulation for normal kinetochore homeostasis.

Cell division in two large pennate diatoms Hantzschia and Nitzschia III. A new proposal for kinetochore function during prometaphase

PubMed Central

1980-01-01

Prometaphase in two large species of diatoms is examined, using the following techniques: (a) time-lapse cinematography of chromosome movements in vivo; (b) electron microscopy of corresponding stages: (c) reconstruction of the microtubules (MTs) in the kinetochore fiber of chromosomes attached to the spindle. In vivo, the chromosomes independently commence oscillations back and forth to one pole. The kinetochore is usually at the leading edge of such chromosome movements; a variable time later both kinetochores undergo such oscillations but toward opposite poles and soon stretch poleward to establish stable bipolar attachment. Electron microscopy of early prometaphase shows that the kinetochores usually laterally associate with MTs that have one end attached to the spindle pole. At late prometaphase, most chromosomes are fully attached to the spindle, but the kinetochores on unattached chromosomes are bare of MTs. Reconstruction of the kinetochore fiber demonstrates that most of its MTs (96%) extend past the kinetochore and are thus apparently not nucleated there. At least one MT terminates at each kinetochore analyzed. Our interpretation is that the conventional view of kinetochore function cannot apply to diatoms. The kinetochore fiber in diatoms appears to be primarily composed of MTs from the poles, in contrast to the conventional view that many MTs of the kinetochore fiber are nucleated by the kinetochore. Similarly, chromosomes appear to initially orient their kinetochores to opposite poles by moving along MTs attached to the poles, instead of orientation effected by kinetochore MTs laterally associating with other MTs in the spindle. The function of the kinetochore in diatoms and other cell types is discussed. PMID:7400213

Mad1 kinetochore recruitment by Mps1-mediated phosphorylation of Bub1 signals the spindle checkpoint.

PubMed

London, Nitobe; Biggins, Sue

2014-01-15

The spindle checkpoint is a conserved signaling pathway that ensures genomic integrity by preventing cell division when chromosomes are not correctly attached to the spindle. Checkpoint activation depends on the hierarchical recruitment of checkpoint proteins to generate a catalytic platform at the kinetochore. Although Mad1 kinetochore localization is the key regulatory downstream event in this cascade, its receptor and mechanism of recruitment have not been conclusively identified. Here, we demonstrate that Mad1 kinetochore association in budding yeast is mediated by phosphorylation of a region within the Bub1 checkpoint protein by the conserved protein kinase Mps1. Tethering this region of Bub1 to kinetochores bypasses the checkpoint requirement for Mps1-mediated kinetochore recruitment of upstream checkpoint proteins. The Mad1 interaction with Bub1 and kinetochores can be reconstituted in the presence of Mps1 and Mad2. Together, this work reveals a critical mechanism that determines kinetochore activation of the spindle checkpoint.

Chk1 and Mps1 jointly regulate correction of merotelic kinetochore attachments.

PubMed

Petsalaki, Eleni; Zachos, George

2013-03-01

If uncorrected, merotelic kinetochore attachments can induce mis-segregated chromosomes in anaphase. We show that checkpoint kinase 1 (Chk1) protects vertebrate cells against merotelic attachments and lagging chromosomes and is required for correction of merotelic attachments during a prolonged metaphase. Decreased Chk1 activity leads to hyper-stable kinetochore microtubules, unstable binding of MCAK, Kif2b and Mps1 to centromeres or kinetochores and reduced phosphorylation of Hec1 by Aurora-B. Phosphorylation of Aurora-B at serine 331 (Ser331) by Chk1 is high in prometaphase and decreases significantly in metaphase cells. We propose that Ser331 phosphorylation is required for optimal localization of MCAK, Kif2b and Mps1 to centromeres or kinetochores and for Hec1 phosphorylation. Furthermore, inhibition of Mps1 activity diminishes initial recruitment of MCAK and Kif2b to centromeres or kinetochores, impairs Hec1 phosphorylation and exacerbates merotelic attachments in Chk1-deficient cells. We propose that Chk1 and Mps1 jointly regulate Aurora-B, MCAK, Kif2b and Hec1 to correct merotelic attachments. These results suggest a role for Chk1 and Mps1 in error correction.

Stable kinetochore-microtubule attachment is sufficient to silence the spindle assembly checkpoint in human cells.

PubMed

Tauchman, Eric C; Boehm, Frederick J; DeLuca, Jennifer G

2015-12-01

During mitosis, duplicated sister chromatids attach to microtubules emanating from opposing sides of the bipolar spindle through large protein complexes called kinetochores. In the absence of stable kinetochore-microtubule attachments, a cell surveillance mechanism known as the spindle assembly checkpoint (SAC) produces an inhibitory signal that prevents anaphase onset. Precisely how the inhibitory SAC signal is extinguished in response to microtubule attachment remains unresolved. To address this, we induced formation of hyper-stable kinetochore-microtubule attachments in human cells using a non-phosphorylatable version of the protein Hec1, a core component of the attachment machinery. We find that stable attachments are sufficient to silence the SAC in the absence of sister kinetochore bi-orientation and strikingly in the absence of detectable microtubule pulling forces or tension. Furthermore, we find that SAC satisfaction occurs despite the absence of large changes in intra-kinetochore distance, suggesting that substantial kinetochore stretching is not required for quenching the SAC signal.

A Coordinated Interdependent Protein Circuitry Stabilizes the Kinetochore Ensemble to Protect CENP-A in the Human Pathogenic Yeast Candida albicans

PubMed Central

Thakur, Jitendra; Sanyal, Kaustuv

2012-01-01

Unlike most eukaryotes, a kinetochore is fully assembled early in the cell cycle in budding yeasts Saccharomyces cerevisiae and Candida albicans. These kinetochores are clustered together throughout the cell cycle. Kinetochore assembly on point centromeres of S. cerevisiae is considered to be a step-wise process that initiates with binding of inner kinetochore proteins on specific centromere DNA sequence motifs. In contrast, kinetochore formation in C. albicans, that carries regional centromeres of 3–5 kb long, has been shown to be a sequence independent but an epigenetically regulated event. In this study, we investigated the process of kinetochore assembly/disassembly in C. albicans. Localization dependence of various kinetochore proteins studied by confocal microscopy and chromatin immunoprecipitation (ChIP) assays revealed that assembly of a kinetochore is a highly coordinated and interdependent event. Partial depletion of an essential kinetochore protein affects integrity of the kinetochore cluster. Further protein depletion results in complete collapse of the kinetochore architecture. In addition, GFP-tagged kinetochore proteins confirmed similar time-dependent disintegration upon gradual depletion of an outer kinetochore protein (Dam1). The loss of integrity of a kinetochore formed on centromeric chromatin was demonstrated by reduced binding of CENP-A and CENP-C at the centromeres. Most strikingly, Western blot analysis revealed that gradual depletion of any of these essential kinetochore proteins results in concomitant reduction in cellular protein levels of CENP-A. We further demonstrated that centromere bound CENP-A is protected from the proteosomal mediated degradation. Based on these results, we propose that a coordinated interdependent circuitry of several evolutionarily conserved essential kinetochore proteins ensures integrity of a kinetochore formed on the foundation of CENP-A containing centromeric chromatin. PMID:22536162

Contributions of Microtubule Dynamic Instability and Rotational Diffusion to Kinetochore Capture.

PubMed

Blackwell, Robert; Sweezy-Schindler, Oliver; Edelmaier, Christopher; Gergely, Zachary R; Flynn, Patrick J; Montes, Salvador; Crapo, Ammon; Doostan, Alireza; McIntosh, J Richard; Glaser, Matthew A; Betterton, Meredith D

2017-02-07

Microtubule dynamic instability allows search and capture of kinetochores during spindle formation, an important process for accurate chromosome segregation during cell division. Recent work has found that microtubule rotational diffusion about minus-end attachment points contributes to kinetochore capture in fission yeast, but the relative contributions of dynamic instability and rotational diffusion are not well understood. We have developed a biophysical model of kinetochore capture in small fission-yeast nuclei using hybrid Brownian dynamics/kinetic Monte Carlo simulation techniques. With this model, we have studied the importance of dynamic instability and microtubule rotational diffusion for kinetochore capture, both to the lateral surface of a microtubule and at or near its end. Over a range of biologically relevant parameters, microtubule rotational diffusion decreased capture time, but made a relatively small contribution compared to dynamic instability. At most, rotational diffusion reduced capture time by 25%. Our results suggest that while microtubule rotational diffusion can speed up kinetochore capture, it is unlikely to be the dominant physical mechanism for typical conditions in fission yeast. In addition, we found that when microtubules undergo dynamic instability, lateral captures predominate even in the absence of rotational diffusion. Counterintuitively, adding rotational diffusion to a dynamic microtubule increases the probability of end-on capture. Copyright © 2017 Biophysical Society. Published by Elsevier Inc. All rights reserved.

Epigenetic engineering: histone H3K9 acetylation is compatible with kinetochore structure and function.

PubMed

Bergmann, Jan H; Jakubsche, Julia N; Martins, Nuno M; Kagansky, Alexander; Nakano, Megumi; Kimura, Hiroshi; Kelly, David A; Turner, Bryan M; Masumoto, Hiroshi; Larionov, Vladimir; Earnshaw, William C

2012-01-15

Human kinetochores are transcriptionally active, producing very low levels of transcripts of the underlying alpha-satellite DNA. However, it is not known whether kinetochores can tolerate acetylated chromatin and the levels of transcription that are characteristic of housekeeping genes, or whether kinetochore-associated 'centrochromatin', despite being transcribed at a low level, is essentially a form of repressive chromatin. Here, we have engineered two types of acetylated chromatin within the centromere of a synthetic human artificial chromosome. Tethering a minimal NF-κB p65 activation domain within kinetochore-associated chromatin produced chromatin with high levels of histone H3 acetylated on lysine 9 (H3K9ac) and an ~10-fold elevation in transcript levels, but had no substantial effect on kinetochore assembly or function. By contrast, tethering the herpes virus VP16 activation domain produced similar modifications in the chromatin but resulted in an ~150-fold elevation in transcripts, approaching the level of transcription of an endogenous housekeeping gene. This rapidly inactivated kinetochores, causing a loss of assembled CENP-A and blocking further CENP-A assembly. Our data reveal that functional centromeres in vivo show a remarkable plasticity--kinetochores tolerate profound changes to their chromatin environment, but appear to be critically sensitive to the level of centromeric transcription.

The kinetochore prevents centromere-proximal crossover recombination during meiosis

PubMed Central

Vincenten, Nadine; Kuhl, Lisa-Marie; Lam, Isabel; Oke, Ashwini; Kerr, Alastair RW; Hochwagen, Andreas; Fung, Jennifer; Keeney, Scott; Vader, Gerben; Marston, Adèle L

2015-01-01

During meiosis, crossover recombination is essential to link homologous chromosomes and drive faithful chromosome segregation. Crossover recombination is non-random across the genome, and centromere-proximal crossovers are associated with an increased risk of aneuploidy, including Trisomy 21 in humans. Here, we identify the conserved Ctf19/CCAN kinetochore sub-complex as a major factor that minimizes potentially deleterious centromere-proximal crossovers in budding yeast. We uncover multi-layered suppression of pericentromeric recombination by the Ctf19 complex, operating across distinct chromosomal distances. The Ctf19 complex prevents meiotic DNA break formation, the initiating event of recombination, proximal to the centromere. The Ctf19 complex independently drives the enrichment of cohesin throughout the broader pericentromere to suppress crossovers, but not DNA breaks. This non-canonical role of the kinetochore in defining a chromosome domain that is refractory to crossovers adds a new layer of functionality by which the kinetochore prevents the incidence of chromosome segregation errors that generate aneuploid gametes. DOI: http://dx.doi.org/10.7554/eLife.10850.001 PMID:26653857

A Taz1- and Microtubule-Dependent Regulatory Relationship between Telomere and Centromere Positions in Bouquet Formation Secures Proper Meiotic Divisions

PubMed Central

Katsumata, Kazuhiro; Hirayasu, Ami; Miyoshi, Junpei; Nishi, Eriko; Ichikawa, Kento; Tateho, Kazuki; Wakuda, Airi; Matsuhara, Hirotada; Yamamoto, Ayumu

2016-01-01

During meiotic prophase, telomeres cluster, forming the bouquet chromosome arrangement, and facilitate homologous chromosome pairing. In fission yeast, bouquet formation requires switching of telomere and centromere positions. Centromeres are located at the spindle pole body (SPB) during mitotic interphase, and upon entering meiosis, telomeres cluster at the SPB, followed by centromere detachment from the SPB. Telomere clustering depends on the formation of the microtubule-organizing center at telomeres by the linker of nucleoskeleton and cytoskeleton complex (LINC), while centromere detachment depends on disassembly of kinetochores, which induces meiotic centromere formation. However, how the switching of telomere and centromere positions occurs during bouquet formation is not fully understood. Here, we show that, when impaired telomere interaction with the LINC or microtubule disruption inhibited telomere clustering, kinetochore disassembly-dependent centromere detachment and accompanying meiotic centromere formation were also inhibited. Efficient centromere detachment required telomere clustering-dependent SPB recruitment of a conserved telomere component, Taz1, and microtubules. Furthermore, when artificial SPB recruitment of Taz1 induced centromere detachment in telomere clustering-defective cells, spindle formation was impaired. Thus, detachment of centromeres from the SPB without telomere clustering causes spindle impairment. These findings establish novel regulatory mechanisms, which prevent concurrent detachment of telomeres and centromeres from the SPB during bouquet formation and secure proper meiotic divisions. PMID:27611693

Dynamic localization of Mps1 kinase to kinetochores is essential for accurate spindle microtubule attachment

PubMed Central

Dou, Zhen; Liu, Xing; Wang, Wenwen; Zhu, Tongge; Wang, Xinghui; Xu, Leilei; Abrieu, Ariane; Fu, Chuanhai; Hill, Donald L.; Yao, Xuebiao

2015-01-01

The spindle assembly checkpoint (SAC) is a conserved signaling pathway that monitors faithful chromosome segregation during mitosis. As a core component of SAC, the evolutionarily conserved kinase monopolar spindle 1 (Mps1) has been implicated in regulating chromosome alignment, but the underlying molecular mechanism remains unclear. Our molecular delineation of Mps1 activity in SAC led to discovery of a previously unidentified structural determinant underlying Mps1 function at the kinetochores. Here, we show that Mps1 contains an internal region for kinetochore localization (IRK) adjacent to the tetratricopeptide repeat domain. Importantly, the IRK region determines the kinetochore localization of inactive Mps1, and an accumulation of inactive Mps1 perturbs accurate chromosome alignment and mitotic progression. Mechanistically, the IRK region binds to the nuclear division cycle 80 complex (Ndc80C), and accumulation of inactive Mps1 at the kinetochores prevents a dynamic interaction between Ndc80C and spindle microtubules (MTs), resulting in an aberrant kinetochore attachment. Thus, our results present a previously undefined mechanism by which Mps1 functions in chromosome alignment by orchestrating Ndc80C–MT interactions and highlight the importance of the precise spatiotemporal regulation of Mps1 kinase activity and kinetochore localization in accurate mitotic progression. PMID:26240331

Dynamic localization of Mps1 kinase to kinetochores is essential for accurate spindle microtubule attachment.

PubMed

Dou, Zhen; Liu, Xing; Wang, Wenwen; Zhu, Tongge; Wang, Xinghui; Xu, Leilei; Abrieu, Ariane; Fu, Chuanhai; Hill, Donald L; Yao, Xuebiao

2015-08-18

The spindle assembly checkpoint (SAC) is a conserved signaling pathway that monitors faithful chromosome segregation during mitosis. As a core component of SAC, the evolutionarily conserved kinase monopolar spindle 1 (Mps1) has been implicated in regulating chromosome alignment, but the underlying molecular mechanism remains unclear. Our molecular delineation of Mps1 activity in SAC led to discovery of a previously unidentified structural determinant underlying Mps1 function at the kinetochores. Here, we show that Mps1 contains an internal region for kinetochore localization (IRK) adjacent to the tetratricopeptide repeat domain. Importantly, the IRK region determines the kinetochore localization of inactive Mps1, and an accumulation of inactive Mps1 perturbs accurate chromosome alignment and mitotic progression. Mechanistically, the IRK region binds to the nuclear division cycle 80 complex (Ndc80C), and accumulation of inactive Mps1 at the kinetochores prevents a dynamic interaction between Ndc80C and spindle microtubules (MTs), resulting in an aberrant kinetochore attachment. Thus, our results present a previously undefined mechanism by which Mps1 functions in chromosome alignment by orchestrating Ndc80C-MT interactions and highlight the importance of the precise spatiotemporal regulation of Mps1 kinase activity and kinetochore localization in accurate mitotic progression.

Kinetochore fiber formation in animal somatic cells: dueling mechanisms come to a draw

PubMed Central

2008-01-01

The attachment to and movement of a chromosome on the mitotic spindle are mediated by the formation of a bundle of microtubules (MTs) that tethers the kinetochore on the chromosome to a spindle pole. The origin of these “kinetochore fibers” (K fibers) has been investigated for over 125 years. As noted in 1944 by Schrader [Mitosis, Columbia University Press, New York, 110 pp.], there are three possible ways to form a K fiber: (a) it grows from the pole until it contacts the kinetochore, (b) it grows directly from the kinetochore, or (c) it forms as a result of an interaction between the pole and the chromosome. Since Schrader's time, it has been firmly established that K fibers in centrosome-containing animal somatic cells form as kinetochores capture MTs growing from the spindle pole (route a). It is now similarly clear that in cells lacking centrosomes, including higher plants and many animal oocytes, K fibers “self-assemble” from MTs generated by the chromosomes (route b). Can animal somatic cells form K fibers in the absence of centrosomes by the “self-assembly” pathway? In 2000, the answer to this question was shown to be a resounding “yes.” With this result, the next question became whether the presence of a centrosome normally suppresses K fiber self-assembly or if this route works concurrently with centrosome-mediated K-fiber formation. This question, too, has recently been answered: observations on untreated live animal cells expressing green fluorescent protein-tagged tubulin clearly show that kinetochores can nucleate the formation of their associated MTs in a unique manner in the presence of functional centrosomes. The concurrent operation of these two “dueling” routes for forming K fibers in animal cells helps explain why the attachment of kinetochores and the maturation of K fibers occur as quickly as they do on all chromosomes within a cell. PMID:16270218

Kinetochore identification in micronuclei in mouse bone-marrow erythrocytes: an assay for the detection of aneuploidy-inducing agents.

PubMed

Gudi, R; Sandhu, S S; Athwal, R S

1990-10-01

An in vivo micronucleus assay using mouse bone marrow for identifying the ability of chemicals to induce aneuploidy and/or chromosome breaks is described. Micronucleus formation in bone-marrow erythrocytes of mice is commonly used as an index for evaluating the clastogenicity of environmental agents. However, micronuclei may also originate from intact lagging chromosomes resulting from the effect of aneuploidy-inducing agents. We have used immunofluorescent staining using anti-kinetochore antibodies to classify micronuclei for the presence or absence of kinetochores. Micronuclei positive for kinetochores are assumed to contain intact chromosomes and result from induced aneuploidy; while those negative for kinetochores contain acentric chromosomal fragments and originate from clastogenic events. The assay was evaluated using X-irradiation (a known clastogen) and vincristine sulfate (an aneuploidy-inducing agent). A dose-related response for the induction of micronuclei was observed for both agents. Micronuclei induced by X-irradiation were negative for kinetochores while the majority of the micronuclei resulting from vincristine treatment contained kinetochores. Thus, the micronucleus assay in combination with immunofluorescent staining for kinetochores may provide a useful method to simultaneously assess the ability of chemicals to induce aneuploidy and/or chromosome breaks.

A Bir1p–Sli15p Kinetochore Passenger Complex Regulates Septin Organization during Anaphase

PubMed Central

Thomas, Scott

2007-01-01

Kinetochore–passenger complexes in metazoans have been proposed to coordinate the segregation of chromosomes in anaphase with the induction of cytokinesis. Passenger protein homologues in the budding yeast Saccharomyces cerevisiae play a critical role early in mitosis, ensuring proper biorientation of kinetochore–microtubule attachments. Our recent work has implicated the passenger protein Bir1p (Survivin) and the inner kinetochore complex centromere binding factor 3 (CBF3) in the regulation of septin dynamics during anaphase. Here, we present data that is consistent with there being multiple passenger protein complexes. Our data show that Bir1p links together a large passenger complex containing Ndc10p, Sli15p (INCENP), and Ipl1p (Aurora B) and that the interaction between Bir1p and Sli15p is specifically involved in regulating septin dynamics during anaphase. Neither conditional alleles nor mutants of BIR1 that disrupt the interaction between Bir1p and Sli15p resulted in mono-attached kinetochores, suggesting that the Bir1p–Sli15p complex functions in anaphase and independently from Sli15p–Ipl1p complexes. We present a model for how discrete passenger complexes coordinate distinct aspects of mitosis. PMID:17652458

UV-C irradiation delays mitotic progression by recruiting Mps1 to kinetochores.

PubMed

Zhang, Xiaojuan; Ling, Youguo; Wang, Wenjun; Zhang, Yanhong; Ma, Qingjun; Tan, Pingping; Song, Ting; Wei, Congwen; Li, Ping; Liu, Xuedong; Ma, Runlin Z; Zhong, Hui; Cao, Cheng; Xu, Quanbin

2013-04-15

The effect of UV irradiation on replicating cells during interphase has been studied extensively. However, how the mitotic cell responds to UV irradiation is less well defined. Herein, we found that UV-C irradiation (254 nm) increases recruitment of the spindle checkpoint proteins Mps1 and Mad2 to the kinetochore during metaphase, suggesting that the spindle assembly checkpoint (SAC) is reactivated. In accordance with this, cells exposed to UV-C showed delayed mitotic progression, characterized by a prolonged chromosomal alignment during metaphase. UV-C irradiation also induced the DNA damage response and caused a significant accumulation of γ-H2AX on mitotic chromosomes. Unexpectedly, the mitotic delay upon UV-C irradiation is not due to the DNA damage response but to the relocation of Mps1 to the kinetochore. Further, we found that UV-C irradiation activates Aurora B kinase. Importantly, the kinase activity of Aurora B is indispensable for full recruitment of Mps1 to the kinetochore during both prometaphase and metaphase. Taking these findings together, we propose that UV irradiation delays mitotic progression by evoking the Aurora B-Mps1 signaling cascade, which exerts its role through promoting the association of Mps1 with the kinetochore in metaphase.

UV-C irradiation delays mitotic progression by recruiting Mps1 to kinetochores

PubMed Central

Zhang, Xiaojuan; Ling, Youguo; Wang, Wenjun; Zhang, Yanhong; Ma, Qingjun; Tan, Pingping; Song, Ting; Wei, Congwen; Li, Ping; Liu, Xuedong; Ma, Runlin Z.; Zhong, Hui; Cao, Cheng; Xu, Quanbin

2013-01-01

The effect of UV irradiation on replicating cells during interphase has been studied extensively. However, how the mitotic cell responds to UV irradiation is less well defined. Herein, we found that UV-C irradiation (254 nm) increases recruitment of the spindle checkpoint proteins Mps1 and Mad2 to the kinetochore during metaphase, suggesting that the spindle assembly checkpoint (SAC) is reactivated. In accordance with this, cells exposed to UV-C showed delayed mitotic progression, characterized by a prolonged chromosomal alignment during metaphase. UV-C irradiation also induced the DNA damage response and caused a significant accumulation of γ-H2AX on mitotic chromosomes. Unexpectedly, the mitotic delay upon UV-C irradiation is not due to the DNA damage response but to the relocation of Mps1 to the kinetochore. Further, we found that UV-C irradiation activates Aurora B kinase. Importantly, the kinase activity of Aurora B is indispensable for full recruitment of Mps1 to the kinetochore during both prometaphase and metaphase. Taking these findings together, we propose that UV irradiation delays mitotic progression by evoking the Aurora B-Mps1 signaling cascade, which exerts its role through promoting the association of Mps1 with the kinetochore in metaphase. PMID:23531678

TRAMM/TrappC12 plays a role in chromosome congression, kinetochore stability, and CENP-E recruitment

PubMed Central

Milev, Miroslav P.; Hasaj, Benedeta; Saint-Dic, Djenann; Snounou, Sary; Zhao, Qingchuan

2015-01-01

Chromosome congression requires the stable attachment of microtubules to chromosomes mediated by the kinetochore, a large proteinaceous structure whose mechanism of assembly is unknown. In this paper, we present the finding that a protein called TRAMM (formerly known as TrappC12) plays a role in mitosis. Depletion of TRAMM resulted in noncongressed chromosomes and arrested cells in mitosis. Small amounts of TRAMM associated with chromosomes, and its depletion affected the localization of some kinetochore proteins, the strongest effect being seen for CENP-E. TRAMM interacts with CENP-E, and depletion of TRAMM prevented the recruitment of CENP-E to the kinetochore. TRAMM is phosphorylated early in mitosis and dephosphorylated at the onset of anaphase. Interestingly, this phosphorylation/dephosphorylation cycle correlates with its association/disassociation with CENP-E. Finally, we demonstrate that a phosphomimetic form of TRAMM recruited CENP-E to kinetochores more efficiently than did the nonphosphorylatable mutant. Our study identifies a moonlighting function for TRAMM during mitosis and adds a new component that regulates kinetochore stability and CENP-E recruitment. PMID:25918224

Distinct Sequence Elements of Cyclin B1 Promote Localization to Chromatin, Centrosomes, and Kinetochores during Mitosis

PubMed Central

Bentley, Anna M.; Normand, Guillaume; Hoyt, Jonathan

2007-01-01

The mitotic cyclins promote cell division by binding and activating cyclin-dependent kinases (CDKs). Each cyclin has a unique pattern of subcellular localization that plays a vital role in regulating cell division. During mitosis, cyclin B1 is known to localize to centrosomes, microtubules, and chromatin. To determine the mechanisms of cyclin B1 localization in M phase, we imaged full-length and mutant versions of human cyclin B1-enhanced green fluorescent protein in live cells by using spinning disk confocal microscopy. In addition to centrosome, microtubule, and chromatin localization, we found that cyclin B1 also localizes to unattached kinetochores after nuclear envelope breakdown. Kinetochore recruitment of cyclin B1 required the kinetochore proteins Hec1 and Mad2, and it was stimulated by microtubule destabilization. Mutagenesis studies revealed that cyclin B1 is recruited to kinetochores through both CDK1-dependent and -independent mechanisms. In contrast, localization of cyclin B1 to chromatin and centrosomes is independent of CDK1 binding. The N-terminal domain of cyclin B1 is necessary and sufficient for chromatin association, whereas centrosome recruitment relies on sequences within the cyclin box. Our data support a role for cyclin B1 function at unattached kinetochores, and they demonstrate that separable and distinct sequence elements target cyclin B1 to kinetochores, chromatin, and centrosomes during mitosis. PMID:17881737

Kinetochore Fiber Maturation in PtK1 Cells and Its Implications for the Mechanisms of Chromosome Congression and Anaphase Onset

PubMed Central

McEwen, Bruce F.; Heagle, Amy B.; Cassels, Grisel O.; Buttle, Karolyn F.; Rieder, Conly L.

1997-01-01

Kinetochore microtubules (kMts) are a subset of spindle microtubules that bind directly to the kinetochore to form the kinetochore fiber (K-fiber). The K-fiber in turn interacts with the kinetochore to produce chromosome motion toward the attached spindle pole. We have examined K-fiber maturation in PtK1 cells using same-cell video light microscopy/serial section EM. During congression, the kinetochore moving away from its spindle pole (i.e., the trailing kinetochore) and its leading, poleward moving sister both have variable numbers of kMts, but the trailing kinetochore always has at least twice as many kMts as the leading kinetochore. A comparison of Mt numbers on sister kinetochores of congressing chromosomes with their direction of motion, as well as distance from their associated spindle poles, reveals that the direction of motion is not determined by kMt number or total kMt length. The same result was observed for oscillating metaphase chromosomes. These data demonstrate that the tendency of a kinetochore to move poleward is not positively correlated with the kMt number. At late prometaphase, the average number of Mts on fully congressed kinetochores is 19.7 ± 6.7 (n = 94), at late metaphase 24.3 ± 4.9 (n = 62), and at early anaphase 27.8 ± 6.3 (n = 65). Differences between these distributions are statistically significant. The increased kMt number during early anaphase, relative to late metaphase, reflects the increased kMt stability at anaphase onset. Treatment of late metaphase cells with 1 μM taxol inhibits anaphase onset, but produces the same kMt distribution as in early anaphase: 28.7 ± 7.4 (n = 54). Thus, a full complement of kMts is not sufficient to induce anaphase onset. We also measured the time course for kMt acquisition and determined an initial rate of 1.9 kMts/min. This rate accelerates up to 10-fold during the course of K-fiber maturation, suggesting an increased concentration of Mt plus ends in the vicinity of the kinetochore at late

Bub1 autophosphorylation feeds back to regulate kinetochore docking and promote localized substrate phosphorylation.

PubMed

Asghar, Adeel; Lajeunesse, Audrey; Dulla, Kalyan; Combes, Guillaume; Thebault, Philippe; Nigg, Erich A; Elowe, Sabine

2015-09-24

During mitosis, Bub1 kinase phosphorylates histone H2A-T120 to promote centromere sister chromatid cohesion through recruitment of shugoshin (Sgo) proteins. The regulation and dynamics of H2A-T120 phosphorylation are poorly understood. Using quantitative phosphoproteomics we show that Bub1 is autophosphorylated at numerous sites. We confirm mitosis-specific autophosphorylation of a several residues and show that Bub1 activation is primed in interphase but fully achieved only in mitosis. Mutation of a single autophosphorylation site T589 alters kinetochore turnover of Bub1 and results in uniform H2A-T120 phosphorylation and Sgo recruitment along chromosome arms. Consequently, improper sister chromatid resolution and chromosome segregation errors are observed. Kinetochore tethering of Bub1-T589A refocuses H2A-T120 phosphorylation and Sgo1 to centromeres. Recruitment of the Bub1-Bub3-BubR1 axis to kinetochores has recently been extensively studied. Our data provide novel insight into the regulation and kinetochore residency of Bub1 and indicate that its localization is dynamic and tightly controlled through feedback autophosphorylation.

Spatiotemporal dynamics of Aurora B-PLK1-MCAK signaling axis orchestrates kinetochore bi-orientation and faithful chromosome segregation

PubMed Central

Shao, Hengyi; Huang, Yuejia; Zhang, Liangyu; Yuan, Kai; Chu, Youjun; Dou, Zhen; Jin, Changjiang; Garcia-Barrio, Minerva; Liu, Xing; Yao, Xuebiao

2015-01-01

Chromosome segregation in mitosis is orchestrated by the dynamic interactions between the kinetochore and spindle microtubules. The microtubule depolymerase mitotic centromere-associated kinesin (MCAK) is a key regulator for an accurate kinetochore-microtubule attachment. However, the regulatory mechanism underlying precise MCAK depolymerase activity control during mitosis remains elusive. Here, we describe a novel pathway involving an Aurora B-PLK1 axis for regulation of MCAK activity in mitosis. Aurora B phosphorylates PLK1 on Thr210 to activate its kinase activity at the kinetochores during mitosis. Aurora B-orchestrated PLK1 kinase activity was examined in real-time mitosis using a fluorescence resonance energy transfer-based reporter and quantitative analysis of native PLK1 substrate phosphorylation. Active PLK1, in turn, phosphorylates MCAK at Ser715 which promotes its microtubule depolymerase activity essential for faithful chromosome segregation. Importantly, inhibition of PLK1 kinase activity or expression of a non-phosphorylatable MCAK mutant prevents correct kinetochore-microtubule attachment, resulting in abnormal anaphase with chromosome bridges. We reason that the Aurora B-PLK1 signaling at the kinetochore orchestrates MCAK activity, which is essential for timely correction of aberrant kinetochore attachment to ensure accurate chromosome segregation during mitosis. PMID:26206521

Zwint-1 is required for spindle assembly checkpoint function and kinetochore-microtubule attachment during oocyte meiosis.

PubMed

Woo Seo, Dong; Yeop You, Seung; Chung, Woo-Jae; Cho, Dong-Hyung; Kim, Jae-Sung; Su Oh, Jeong

2015-10-21

The key step for faithful chromosome segregation during meiosis is kinetochore assembly. Defects in this process result in aneuploidy, leading to miscarriages, infertility and various birth defects. However, the roles of kinetochores in homologous chromosome segregation during meiosis are ill-defined. Here we found that Zwint-1 is required for homologous chromosome segregation during meiosis. Knockdown of Zwint-1 accelerated the first meiosis by abrogating the kinetochore recruitment of Mad2, leading to chromosome misalignment and a high incidence of aneuploidy. Although Zwint-1 knockdown did not affect Aurora C kinase activity, the meiotic defects following Zwint-1 knockdown were similar to those observed with ZM447439 treatment. Importantly, the chromosome misalignment following Aurora C kinase inhibition was not restored after removing the inhibitor in Zwint-1-knockdown oocytes, whereas the defect was rescued after the inhibitor washout in the control oocytes. These results suggest that Aurora C kinase-mediated correction of erroneous kinetochore-microtubule attachment is primarily regulated by Zwint-1. Our results provide the first evidence that Zwint-1 is required to correct erroneous kinetochore-microtubule attachment and regulate spindle checkpoint function during meiosis.

SKAP, an outer kinetochore protein, is required for mouse germ cell development

PubMed Central

Grey, Corinne; Espeut, Julien; Ametsitsi, Rachel; Kumar, Rajeev; Luksza, Malgorzata; Brun, Christine; Verlhac, Marie-Hélene; Suja, José Angél; de Massy, Bernard

2016-01-01

In sexually reproducing organisms, accurate gametogenesis is crucial for the transmission of genetic material from one generation to the next. This requires the faithful segregation of chromosomes during mitotic and meiotic divisions. One of the main players in this process is the kinetochore, a large multi-protein complex that forms at the interface of centromeres and microtubules. Here, we analyzed the expression profile and function of small kinetochore-associated protein (SKAP) in the mouse. We found that two distinct SKAP isoforms are specifically expressed in the germline: a smaller isoform, which is detected in spermatogonia and spermatocytes and localized in the outer mitotic and meiotic kinetochores from metaphase to telophase, and a larger isoform, which is expressed in the cytoplasm of elongating spermatids. We generated SKAP-deficient mice and found that testis size and sperm production were severely reduced in mutant males. This phenotype was partially caused by defects during spermatogonia proliferation before entry into meiosis. We conclude that mouse SKAP, while being dispensable for somatic cell divisions, has an important role in the successful outcome of male gametogenesis. In germ cells, analogous to what has been suggested in studies using immortalized cells, SKAP most likely stabilizes the interaction between kinetochores and microtubules, where it might be needed as an extra safeguard to ensure the correct segregation of mitotic and meiotic chromosomes. PMID:26667018

Drosophila Nnf1 paralogs are partially redundant for somatic and germ line kinetochore function.

PubMed

Blattner, Ariane C; Aguilar-Rodríguez, José; Kränzlin, Marcella; Wagner, Andreas; Lehner, Christian F

2017-02-01

Kinetochores allow attachment of chromosomes to spindle microtubules. Moreover, they host proteins that permit correction of erroneous attachments and prevent premature anaphase onset before bi-orientation of all chromosomes in metaphase has been achieved. Kinetochores are assembled from subcomplexes. Kinetochore proteins as well as the underlying centromere proteins and the centromeric DNA sequences evolve rapidly despite their fundamental importance for faithful chromosome segregation during mitotic and meiotic divisions. During evolution of Drosophila melanogaster, several centromere proteins were lost and a recent gene duplication has resulted in two Nnf1 paralogs, Nnf1a and Nnf1b, which code for alternative forms of a Mis12 kinetochore complex component. The rapid evolutionary divergence of centromere/kinetochore constituents in animals and plants has been proposed to be driven by an intragenome conflict resulting from centromere drive during female meiosis. Thus, a female meiosis-specific paralog might be expected to evolve rapidly under positive selection. While our characterization of the D. melanogaster Nnf1 paralogs hints at some partial functional specialization of Nnf1b for meiosis, we have failed to detect evidence for positive selection in our analysis of Nnf1 sequence evolution in the Drosophilid lineage. Neither paralog is essential, even though we find some clear differences in subcellular localization and expression during development. Loss of both paralogs results in developmental lethality. We therefore conclude that the two paralogs are still in early stages of differentiation.

Rad52 phosphorylation by Ipl1 and Mps1 contributes to Mps1 kinetochore localization and spindle assembly checkpoint regulation

PubMed Central

Lim, Gyubum

2017-01-01

Rad52 is well known as a key factor in homologous recombination. Here, we report that Rad52 has functions unrelated to homologous recombination in Saccharomyces cerevisiae; it plays a role in the recruitment of Mps1 to the kinetochores and the maintenance of spindle assembly checkpoint (SAC) activity. Deletion of RAD52 causes various phenotypes related to the dysregulation of chromosome biorientation. Rad52 directly affects efficient operation of the SAC and accurate chromosome segregation. Remarkably, by using an in vitro kinase assay, we found that Rad52 is a substrate of Ipl1/Aurora and Mps1 in yeast and humans. Ipl1-dependent phosphorylation of Rad52 facilitates the kinetochore accumulation of Mps1, and Mps1-dependent phosphorylation of Rad52 is important for the accurate regulation of the SAC under spindle damage conditions. Taken together, our data provide detailed insights into the regulatory mechanism of chromosome biorientation by mitotic kinases. PMID:29078282

Rad52 phosphorylation by Ipl1 and Mps1 contributes to Mps1 kinetochore localization and spindle assembly checkpoint regulation.

PubMed

Lim, Gyubum; Huh, Won-Ki

2017-10-31

Rad52 is well known as a key factor in homologous recombination. Here, we report that Rad52 has functions unrelated to homologous recombination in Saccharomyces cerevisiae ; it plays a role in the recruitment of Mps1 to the kinetochores and the maintenance of spindle assembly checkpoint (SAC) activity. Deletion of RAD52 causes various phenotypes related to the dysregulation of chromosome biorientation. Rad52 directly affects efficient operation of the SAC and accurate chromosome segregation. Remarkably, by using an in vitro kinase assay, we found that Rad52 is a substrate of Ipl1/Aurora and Mps1 in yeast and humans. Ipl1-dependent phosphorylation of Rad52 facilitates the kinetochore accumulation of Mps1, and Mps1-dependent phosphorylation of Rad52 is important for the accurate regulation of the SAC under spindle damage conditions. Taken together, our data provide detailed insights into the regulatory mechanism of chromosome biorientation by mitotic kinases. Published under the PNAS license.

Bub1 autophosphorylation feeds back to regulate kinetochore docking and promote localized substrate phosphorylation

PubMed Central

Asghar, Adeel; Lajeunesse, Audrey; Dulla, Kalyan; Combes, Guillaume; Thebault, Philippe; Nigg, Erich A.; Elowe, Sabine

2015-01-01

During mitosis, Bub1 kinase phosphorylates histone H2A-T120 to promote centromere sister chromatid cohesion through recruitment of shugoshin (Sgo) proteins. The regulation and dynamics of H2A-T120 phosphorylation are poorly understood. Using quantitative phosphoproteomics we show that Bub1 is autophosphorylated at numerous sites. We confirm mitosis-specific autophosphorylation of a several residues and show that Bub1 activation is primed in interphase but fully achieved only in mitosis. Mutation of a single autophosphorylation site T589 alters kinetochore turnover of Bub1 and results in uniform H2A-T120 phosphorylation and Sgo recruitment along chromosome arms. Consequently, improper sister chromatid resolution and chromosome segregation errors are observed. Kinetochore tethering of Bub1-T589A refocuses H2A-T120 phosphorylation and Sgo1 to centromeres. Recruitment of the Bub1-Bub3-BubR1 axis to kinetochores has recently been extensively studied. Our data provide novel insight into the regulation and kinetochore residency of Bub1 and indicate that its localization is dynamic and tightly controlled through feedback autophosphorylation. PMID:26399325

Budding Yeast Kinetochore Proteins, Chl4 and Ctf19, Are Required to Maintain SPB-Centromere Proximity during G1 and Late Anaphase

PubMed Central

Sau, Soumitra; Sutradhar, Sabyasachi; Paul, Raja; Sinha, Pratima

2014-01-01

In the budding yeast, centromeres stay clustered near the spindle pole bodies (SPBs) through most of the cell cycle. This SPB-centromere proximity requires microtubules and functional kinetochores, which are protein complexes formed on the centromeres and capable of binding microtubules. The clustering is suggested by earlier studies to depend also on protein-protein interactions between SPB and kinetochore components. Previously it has been shown that the absence of non-essential kinetochore proteins of the Ctf19 complex weakens kinetochore-microtubule interaction, but whether this compromised interaction affects centromere/kinetochore positioning inside the nucleus is unknown. We found that in G1 and in late anaphase, SPB-centromere proximity was disturbed in mutant cells lacking Ctf19 complex members,Chl4p and/or Ctf19p, whose centromeres lay further away from their SPBs than those of the wild-type cells. We unequivocally show that the SPB-centromere proximity and distances are not dependent on physical interactions between SPB and kinetochore components, but involve microtubule-dependent forces only. Further insight on the positional difference between wild-type and mutant kinetochores was gained by generating computational models governed by (1) independently regulated, but constant kinetochore microtubule (kMT) dynamics, (2) poleward tension on kinetochore and the antagonistic polar ejection force and (3) length and force dependent kMT dynamics. Numerical data obtained from the third model concurs with experimental results and suggests that the absence of Chl4p and/or Ctf19p increases the penetration depth of a growing kMT inside the kinetochore and increases the rescue frequency of a depolymerizing kMT. Both the processes result in increased distance between SPB and centromere. PMID:25003500

Ectopic expression of plasma membrane targeted subunits of the Ndc80-complex as a tool to study kinetochore biochemistry.

PubMed

Holmström, Tim H; Rehnberg, Jonathan; Ahonen, Leena J; Kallio, Marko J

2009-06-01

Genomic stability depends on the normal function of the kinetochore, a multi-protein assemblage, which consists of over 80 molecules including both constitutive and transiently binding components. Information regarding the spatial-temporal assembly of kinetochore subcomplexes is often limited by technical difficulties in their isolation. To study kinetochore subcomplex formation, we targeted separately Hec1 and Spc24, two subunits of the Ndc80 kinetochore compilation, to the plasma membrane by fusing them with the amino-terminal palmitoylation and myristoylation (pm) sequence of the receptor tyrosine kinase Fyn. We found that in early mitotic cells, pm-GFP-Hec1 and pm-GFP-Spc24 fusion proteins localised to the plasma membrane and were able to recruit all subunits of the Ndc80 complex (Ndc80/Hec1, Nuf2, Spc24 and Spc25) to these foci. In interphase cells, only Hec1-Nuf2 and Spc24-Spc25 heterodimers accumulated to the plasma membrane foci. The results propose that the assembly of Ndc80 tetramer can take place outside of the kinetochore but requires co-factors that are only present in mitotic cells. These findings provide the first experimental evidence on the successful employment of the plasma membrane targeting technique in the study of kinetochore biochemistry.

Arabidopsis kinetochore null2 is an upstream component for centromeric histone H3 variant cenH3 deposition at centromeres.

PubMed

Lermontova, Inna; Kuhlmann, Markus; Friedel, Swetlana; Rutten, Twan; Heckmann, Stefan; Sandmann, Michael; Demidov, Dmitri; Schubert, Veit; Schubert, Ingo

2013-09-01

The centromeric histone H3 variant cenH3 is an essential centromeric protein required for assembly, maintenance, and proper function of kinetochores during mitosis and meiosis. We identified a kinetochore null2 (KNL2) homolog in Arabidopsis thaliana and uncovered features of its role in cenH3 loading at centromeres. We show that Arabidopsis KNL2 colocalizes with cenH3 and is associated with centromeres during all stages of the mitotic cell cycle, except from metaphase to mid-anaphase. KNL2 is regulated by the proteasome degradation pathway. The KNL2 promoter is mainly active in meristematic tissues, similar to the cenH3 promoter. A knockout mutant for KNL2 shows a reduced level of cenH3 expression and reduced amount of cenH3 protein at chromocenters of meristematic nuclei, anaphase bridges during mitosis, micronuclei in pollen tetrads, and 30% seed abortion. Moreover, knl2 mutant plants display reduced expression of suppressor of variegation 3-9 homologs2, 4, and 9 and reduced DNA methylation, suggesting an impact of KNL2 on the epigenetic environment for centromere maintenance.

The Mps1 kinase modulates the recruitment and activity of Cnn1(CENP-T) at Saccharomyces cerevisiae kinetochores.

PubMed

Thapa, Kriti Shrestha; Oldani, Amanda; Pagliuca, Cinzia; De Wulf, Peter; Hazbun, Tony R

2015-05-01

Kinetochores are conserved protein complexes that bind the replicated chromosomes to the mitotic spindle and then direct their segregation. To better comprehend Saccharomyces cerevisiae kinetochore function, we dissected the phospho-regulated dynamic interaction between conserved kinetochore protein Cnn1(CENP-T), the centromere region, and the Ndc80 complex through the cell cycle. Cnn1 localizes to kinetochores at basal levels from G1 through metaphase but accumulates abruptly at anaphase onset. How Cnn1 is recruited and which activities regulate its dynamic localization are unclear. We show that Cnn1 harbors two kinetochore-localization activities: a C-terminal histone-fold domain (HFD) that associates with the centromere region and a N-terminal Spc24/Spc25 interaction sequence that mediates linkage to the microtubule-binding Ndc80 complex. We demonstrate that the established Ndc80 binding site in the N terminus of Cnn1, Cnn1(60-84), should be extended with flanking residues, Cnn1(25-91), to allow near maximal binding affinity to Ndc80. Cnn1 localization was proposed to depend on Mps1 kinase activity at Cnn1-S74, based on in vitro experiments demonstrating the Cnn1-Ndc80 complex interaction. We demonstrate that from G1 through metaphase, Cnn1 localizes via both its HFD and N-terminal Spc24/Spc25 interaction sequence, and deletion or mutation of either region results in anomalous Cnn1 kinetochore levels. At anaphase onset (when Mps1 activity decreases) Cnn1 becomes enriched mainly via the N-terminal Spc24/Spc25 interaction sequence. In sum, we provide the first in vivo evidence of Cnn1 preanaphase linkages with the kinetochore and enrichment of the linkages during anaphase. Copyright © 2015 by the Genetics Society of America.

Defining the molecular basis of BubR1 kinetochore interactions and APC/C-CDC20 inhibition.

PubMed

D'Arcy, Sheena; Davies, Owen R; Blundell, Tom L; Bolanos-Garcia, Victor M

2010-05-07

BubR1 is essential for the mitotic checkpoint that prevents aneuploidy in cellular progeny by triggering anaphase delay in response to kinetochores incorrectly/not attached to the mitotic spindle. Here, we define the molecular architecture of the functionally significant N-terminal region of human BubR1 and present the 1.8 A crystal structure of its tetratricopeptide repeat (TPR) domain. The structure reveals divergence from the classical TPR fold and is highly similar to the TPR domain of budding yeast Bub1. Shared distinctive features include a disordered loop insertion, a 3(10)-helix, a tight turn involving glycine positive Phi angles, and noncanonical packing of and between the TPR motifs. We also define the molecular determinants of the interaction between BubR1 and kinetochore protein Blinkin. We identify a shallow groove on the concave surface of the BubR1 TPR domain that forms multiple discrete and potentially cooperative interactions with Blinkin. Finally, we present evidence for a direct interaction between BubR1 and Bub1 mediated by regions C-terminal to their TPR domains. This interaction provides a mechanism for Bub1-dependent kinetochore recruitment of BubR1. We thus present novel molecular insights into the structure of BubR1 and its interactions at the kinetochore-microtubule interface. Our studies pave the way for future structure-directed engineering aimed at dissecting the roles of kinetochore-bound and other pools of BubR1 in vivo.

Phosphorylation of microtubule-binding protein Hec1 by mitotic kinase Aurora B specifies spindle checkpoint kinase Mps1 signaling at the kinetochore.

PubMed

Zhu, Tongge; Dou, Zhen; Qin, Bo; Jin, Changjiang; Wang, Xinghui; Xu, Leilei; Wang, Zhaoyang; Zhu, Lijuan; Liu, Fusheng; Gao, Xinjiao; Ke, Yuwen; Wang, Zhiyong; Aikhionbare, Felix; Fu, Chuanhai; Ding, Xia; Yao, Xuebiao

2013-12-13

The spindle assembly checkpoint (SAC) is a quality control device to ensure accurate chromosome attachment to spindle microtubule for equal segregation of sister chromatid. Aurora B is essential for SAC function by sensing chromosome bi-orientation via spatial regulation of kinetochore substrates. However, it has remained elusive as to how Aurora B couples kinetochore-microtubule attachment to SAC signaling. Here, we show that Hec1 interacts with Mps1 and specifies its kinetochore localization via its calponin homology (CH) domain and N-terminal 80 amino acids. Interestingly, phosphorylation of the Hec1 by Aurora B weakens its interaction with microtubules but promotes Hec1 binding to Mps1. Significantly, the temporal regulation of Hec1 phosphorylation orchestrates kinetochore-microtubule attachment and Mps1 loading to the kinetochore. Persistent expression of phosphomimetic Hec1 mutant induces a hyperactivation of SAC, suggesting that phosphorylation-elicited Hec1 conformational change is used as a switch to orchestrate SAC activation to concurrent destabilization of aberrant kinetochore attachment. Taken together, these results define a novel role for Aurora B-Hec1-Mps1 signaling axis in governing accurate chromosome segregation in mitosis.

Phosphorylation of Microtubule-binding Protein Hec1 by Mitotic Kinase Aurora B Specifies Spindle Checkpoint Kinase Mps1 Signaling at the Kinetochore*

PubMed Central

Zhu, Tongge; Dou, Zhen; Qin, Bo; Jin, Changjiang; Wang, Xinghui; Xu, Leilei; Wang, Zhaoyang; Zhu, Lijuan; Liu, Fusheng; Gao, Xinjiao; Ke, Yuwen; Wang, Zhiyong; Aikhionbare, Felix; Fu, Chuanhai; Ding, Xia; Yao, Xuebiao

2013-01-01

The spindle assembly checkpoint (SAC) is a quality control device to ensure accurate chromosome attachment to spindle microtubule for equal segregation of sister chromatid. Aurora B is essential for SAC function by sensing chromosome bi-orientation via spatial regulation of kinetochore substrates. However, it has remained elusive as to how Aurora B couples kinetochore-microtubule attachment to SAC signaling. Here, we show that Hec1 interacts with Mps1 and specifies its kinetochore localization via its calponin homology (CH) domain and N-terminal 80 amino acids. Interestingly, phosphorylation of the Hec1 by Aurora B weakens its interaction with microtubules but promotes Hec1 binding to Mps1. Significantly, the temporal regulation of Hec1 phosphorylation orchestrates kinetochore-microtubule attachment and Mps1 loading to the kinetochore. Persistent expression of phosphomimetic Hec1 mutant induces a hyperactivation of SAC, suggesting that phosphorylation-elicited Hec1 conformational change is used as a switch to orchestrate SAC activation to concurrent destabilization of aberrant kinetochore attachment. Taken together, these results define a novel role for Aurora B-Hec1-Mps1 signaling axis in governing accurate chromosome segregation in mitosis. PMID:24187132

The Ndc80 internal loop is required for recruitment of the Ska complex to establish end-on microtubule attachment to kinetochores.

PubMed

Zhang, Gang; Kelstrup, Christian D; Hu, Xiao-Wen; Kaas Hansen, Mathilde J; Singleton, Martin R; Olsen, Jesper V; Nilsson, Jakob

2012-07-01

The Ndc80 complex establishes end-on attachment of kinetochores to microtubules, which is essential for chromosome segregation. The Ndc80 subunit is characterized by an N-terminal region that binds directly to microtubules, and a long coiled-coil region that interacts with Nuf2. A loop region in Ndc80 that generates a kink in the structure disrupts the long coiled-coil region but the exact function of this loop, has until now, not been clear. Here we show that this loop region is essential for end-on attachment of kinetochores to microtubules in human cells. Cells expressing loop mutants of Ndc80 are unable to align the chromosomes, and stable kinetochore fibers are absent. Through quantitative mass spectrometry and immunofluorescence we found that the binding of the spindle and kinetochore associated (Ska) complex depends on the loop region, explaining why end-on attachment is defective. This underscores the importance of the Ndc80 loop region in coordinating chromosome segregation through the recruitment of specific proteins to the kinetochore.

Chromosome Bridges Maintain Kinetochore-Microtubule Attachment throughout Mitosis and Rarely Break during Anaphase.

PubMed

Pampalona, Judit; Roscioli, Emanuele; Silkworth, William T; Bowden, Brent; Genescà, Anna; Tusell, Laura; Cimini, Daniela

2016-01-01

Accurate chromosome segregation during cell division is essential to maintain genome stability, and chromosome segregation errors are causally linked to genetic disorders and cancer. An anaphase chromosome bridge is a particular chromosome segregation error observed in cells that enter mitosis with fused chromosomes/sister chromatids. The widely accepted Breakage/Fusion/Bridge cycle model proposes that anaphase chromosome bridges break during mitosis to generate chromosome ends that will fuse during the following cell cycle, thus forming new bridges that will break, and so on. However, various studies have also shown a link between chromosome bridges and aneuploidy and/or polyploidy. In this study, we investigated the behavior and properties of chromosome bridges during mitosis, with the idea to gain insight into the potential mechanism underlying chromosome bridge-induced aneuploidy. We find that only a small number of chromosome bridges break during anaphase, whereas the rest persist through mitosis into the subsequent cell cycle. We also find that the microtubule bundles (k-fibers) bound to bridge kinetochores are not prone to breakage/detachment, thus supporting the conclusion that k-fiber detachment is not the cause of chromosome bridge-induced aneuploidy. Instead, our data suggest that while the microtubules bound to the kinetochores of normally segregating chromosomes shorten substantially during anaphase, the k-fibers bound to bridge kinetochores shorten only slightly, and may even lengthen, during anaphase. This causes some of the bridge kinetochores/chromosomes to lag behind in a position that is proximal to the cell/spindle equator and may cause the bridged chromosomes to be segregated into the same daughter nucleus or to form a micronucleus.

Mutations in CENPE define a novel kinetochore-centromeric mechanism for Microcephalic Primordial Dwarfism

PubMed Central

Mirzaa, Ghayda M.; Vitre, Benjamin; Carpenter, Gillian; Abramowicz, Iga; Gleeson, Joseph G.; Paciorkowski, Alex R.; Cleveland, Don W.; Dobyns, William B.; O’Driscoll, Mark

2015-01-01

Defects in centrosome, centrosomal-associated and spindle-associated proteins are the most frequent cause of Primary Microcephaly (PM) and Microcephalic Primordial Dwarfism (MPD) syndromes in humans. Mitotic progression and segregation defects, microtubule spindle abnormalities and impaired DNA damage-induced G2-M cell cycle checkpoint proficiency have been documented in cell lines from these patients. This suggests that impaired mitotic entry, progression and exit strongly contribute to PM and MPD. Considering the vast protein networks involved in coordinating this cell cycle stage, the list of potential target genes that could underlie novel developmental disorders is large. One such complex network, with a direct microtubule-mediated physical connection to the centrosome, is the kinetochore. This centromeric-associated structure nucleates microtubule attachments onto mitotic chromosomes. Here, we described novel compound heterozygous variants in CENPE in two siblings who exhibit a profound MPD associated with developmental delay, simplified gyri and other isolated abnormalities. CENPE encodes centromere-associated protein E (CENP-E), a core kinetochore component functioning to mediate chromosome congression initially of misaligned chromosomes and in subsequent spindle microtubule capture during mitosis. Firstly, we present a comprehensive clinical description of these patients. Then, using patient cells we document abnormalities in spindle microtubule organisation, mitotic progression and segregation, before modeling the cellular pathogenicity of these variants in an independent cell system. Our cellular analysis shows that a pathogenic defect in CENP-E, a kinetochore-core protein, largely phenocopies PCNT-mutated Microcephalic Osteodysplastic Primordial Dwarfism type II patient cells. PCNT encodes a centrosome-associated protein. These results highlight a common underlying pathomechanism. Our findings provide the first evidence for a kinetochore-based route to

Mutations in CENPE define a novel kinetochore-centromeric mechanism for microcephalic primordial dwarfism.

PubMed

Mirzaa, Ghayda M; Vitre, Benjamin; Carpenter, Gillian; Abramowicz, Iga; Gleeson, Joseph G; Paciorkowski, Alex R; Cleveland, Don W; Dobyns, William B; O'Driscoll, Mark

2014-08-01

Defects in centrosome, centrosomal-associated and spindle-associated proteins are the most frequent cause of primary microcephaly (PM) and microcephalic primordial dwarfism (MPD) syndromes in humans. Mitotic progression and segregation defects, microtubule spindle abnormalities and impaired DNA damage-induced G2-M cell cycle checkpoint proficiency have been documented in cell lines from these patients. This suggests that impaired mitotic entry, progression and exit strongly contribute to PM and MPD. Considering the vast protein networks involved in coordinating this cell cycle stage, the list of potential target genes that could underlie novel developmental disorders is large. One such complex network, with a direct microtubule-mediated physical connection to the centrosome, is the kinetochore. This centromeric-associated structure nucleates microtubule attachments onto mitotic chromosomes. Here, we described novel compound heterozygous variants in CENPE in two siblings who exhibit a profound MPD associated with developmental delay, simplified gyri and other isolated abnormalities. CENPE encodes centromere-associated protein E (CENP-E), a core kinetochore component functioning to mediate chromosome congression initially of misaligned chromosomes and in subsequent spindle microtubule capture during mitosis. Firstly, we present a comprehensive clinical description of these patients. Then, using patient cells we document abnormalities in spindle microtubule organization, mitotic progression and segregation, before modeling the cellular pathogenicity of these variants in an independent cell system. Our cellular analysis shows that a pathogenic defect in CENP-E, a kinetochore-core protein, largely phenocopies PCNT-mutated microcephalic osteodysplastic primordial dwarfism-type II patient cells. PCNT encodes a centrosome-associated protein. These results highlight a common underlying pathomechanism. Our findings provide the first evidence for a kinetochore-based route to

Mps1 Regulates Kinetochore-Microtubule Attachment Stability via the Ska Complex to Ensure Error-Free Chromosome Segregation.

PubMed

Maciejowski, John; Drechsler, Hauke; Grundner-Culemann, Kathrin; Ballister, Edward R; Rodriguez-Rodriguez, Jose-Antonio; Rodriguez-Bravo, Veronica; Jones, Mathew J K; Foley, Emily; Lampson, Michael A; Daub, Henrik; McAinsh, Andrew D; Jallepalli, Prasad V

2017-04-24

The spindle assembly checkpoint kinase Mps1 not only inhibits anaphase but also corrects erroneous attachments that could lead to missegregation and aneuploidy. However, Mps1's error correction-relevant substrates are unknown. Using a chemically tuned kinetochore-targeting assay, we show that Mps1 destabilizes microtubule attachments (K fibers) epistatically to Aurora B, the other major error-correcting kinase. Through quantitative proteomics, we identify multiple sites of Mps1-regulated phosphorylation at the outer kinetochore. Substrate modification was microtubule sensitive and opposed by PP2A-B56 phosphatases that stabilize chromosome-spindle attachment. Consistently, Mps1 inhibition rescued K-fiber stability after depleting PP2A-B56. We also identify the Ska complex as a key effector of Mps1 at the kinetochore-microtubule interface, as mutations that mimic constitutive phosphorylation destabilized K fibers in vivo and reduced the efficiency of the Ska complex's conversion from lattice diffusion to end-coupled microtubule binding in vitro. Our results reveal how Mps1 dynamically modifies kinetochores to correct improper attachments and ensure faithful chromosome segregation. Copyright © 2017 Elsevier Inc. All rights reserved.

Protein Arms in the Kinetochore-Microtubule Interface of the Yeast DASH Complex

PubMed Central

Miranda, JJ L.; King, David S.

2007-01-01

The yeast DASH complex is a heterodecameric component of the kinetochore necessary for accurate chromosome segregation. DASH forms closed rings around microtubules with a large gap between the DASH ring and the microtubule cylinder. We characterized the microtubule-binding properties of limited proteolysis products and subcomplexes of DASH, thus identifying candidate polypeptide extensions involved in establishing the DASH-microtubule interface. The acidic C-terminal extensions of tubulin subunits are not essential for DASH binding. We also measured the molecular mass of DASH rings on microtubules with scanning transmission electron microscopy and found that approximately 25 DASH heterodecamers assemble to form each ring. Dynamic association and relocation of multiple flexible appendages of DASH may allow the kinetochore to translate along the microtubule surface. PMID:17460120

Finding the middle ground: how kinetochores power chromosome congression

PubMed Central

Saurin, Adrian T.

2010-01-01

Genomic stability requires error-free chromosome segregation during mitosis. Chromosome congression to the spindle equator precedes chromosome segregation in anaphase and is a hallmark of metazoan mitosis. Here we review the current knowledge and concepts on the processes that underlie chromosome congression, including initial attachment to spindle microtubules, biorientation, and movements, from the perspective of the kinetochore. PMID:20232224

A TPR domain–containing N-terminal module of MPS1 is required for its kinetochore localization by Aurora B

PubMed Central

Nijenhuis, Wilco; von Castelmur, Eleonore; Littler, Dene; De Marco, Valeria; Tromer, Eelco; Vleugel, Mathijs; van Osch, Maria H.J.; Snel, Berend

2013-01-01

The mitotic checkpoint ensures correct chromosome segregation by delaying cell cycle progression until all kinetochores have attached to the mitotic spindle. In this paper, we show that the mitotic checkpoint kinase MPS1 contains an N-terminal localization module, organized in an N-terminal extension (NTE) and a tetratricopeptide repeat (TPR) domain, for which we have determined the crystal structure. Although the module was necessary for kinetochore localization of MPS1 and essential for the mitotic checkpoint, the predominant kinetochore binding activity resided within the NTE. MPS1 localization further required HEC1 and Aurora B activity. We show that MPS1 localization to kinetochores depended on the calponin homology domain of HEC1 but not on Aurora B–dependent phosphorylation of the HEC1 tail. Rather, the TPR domain was the critical mediator of Aurora B control over MPS1 localization, as its deletion rendered MPS1 localization insensitive to Aurora B inhibition. These data are consistent with a model in which Aurora B activity relieves a TPR-dependent inhibitory constraint on MPS1 localization. PMID:23569217

A TPR domain-containing N-terminal module of MPS1 is required for its kinetochore localization by Aurora B.

PubMed

Nijenhuis, Wilco; von Castelmur, Eleonore; Littler, Dene; De Marco, Valeria; Tromer, Eelco; Vleugel, Mathijs; van Osch, Maria H J; Snel, Berend; Perrakis, Anastassis; Kops, Geert J P L

2013-04-15

The mitotic checkpoint ensures correct chromosome segregation by delaying cell cycle progression until all kinetochores have attached to the mitotic spindle. In this paper, we show that the mitotic checkpoint kinase MPS1 contains an N-terminal localization module, organized in an N-terminal extension (NTE) and a tetratricopeptide repeat (TPR) domain, for which we have determined the crystal structure. Although the module was necessary for kinetochore localization of MPS1 and essential for the mitotic checkpoint, the predominant kinetochore binding activity resided within the NTE. MPS1 localization further required HEC1 and Aurora B activity. We show that MPS1 localization to kinetochores depended on the calponin homology domain of HEC1 but not on Aurora B-dependent phosphorylation of the HEC1 tail. Rather, the TPR domain was the critical mediator of Aurora B control over MPS1 localization, as its deletion rendered MPS1 localization insensitive to Aurora B inhibition. These data are consistent with a model in which Aurora B activity relieves a TPR-dependent inhibitory constraint on MPS1 localization.

Establishing biorientation occurs with precocious separation of the sister kinetochores, but not the arms, in the early spindle of budding yeast.

PubMed

Goshima, G; Yanagida, M

2000-03-17

Sister kinetochores are bioriented toward the spindle poles in higher eukaryotic prometaphase before chromosome segregation. We show that, in budding yeast, the sister kinetochores are separated in the very early spindle, while the sister arms remain associated. Biorientation of the separated kinetochores is achieved already after replication. Mtw1p, a homolog of fission yeast Mis12 required for biorientation, locates at the centromeres in an Ndc10p-dependent manner. Mtw1p and the sequences 1.8 and 3.8 kb from CEN3 and CEN15, respectively, behave like the precociously separated kinetochores, whereas the sequences 23 and 35 kb distant from CEN3 and CEN5 previously used as the centromere markers behave like a part of the arm. Mtw1p and Ndc10p are identically located except for additional spindle localization of Ndc10p. A model explaining small centromeres and early spindle formation in budding yeast is proposed.

Cutting edge science: Laser surgery illuminates viscoelasticity of merotelic kinetochores

PubMed Central

Cabello, Simon

2016-01-01

Increasing evidence in eukaryotic cells suggests that mechanical forces are essential for building a robust mitotic apparatus and correcting inappropriate chromosome attachments. In this issue, Cojoc et al. (2016. J. Cell Biol., http://dx.doi.org/10.1083/jcb.201506011) use laser microsurgery in vivo to measure and study the viscoelastic properties of kinetochores. PMID:27002164

Cutting edge science: Laser surgery illuminates viscoelasticity of merotelic kinetochores.

PubMed

Cabello, Simon; Gachet, Yannick; Tournier, Sylvie

2016-03-28

Increasing evidence in eukaryotic cells suggests that mechanical forces are essential for building a robust mitotic apparatus and correcting inappropriate chromosome attachments. In this issue, Cojoc et al. (2016. J. Cell Biol., http://dx.doi.org/10.1083/jcb.201506011) use laser microsurgery in vivo to measure and study the viscoelastic properties of kinetochores. © 2016 Cabello.

GTPase Ran strongly accumulates at the kinetochores of somatic chromosomes in the spermatogonial mitoses of Acricotopus lucidus (Diptera, Chironomidae).

PubMed

Staiber, Wolfgang

2014-07-01

Unequal chromosome segregation and spindle formation occurs in the last gonial mitosis in the germ line of the chironomid Acricotopus lucidus. During this differential mitosis, all germ line-limited chromosomes (=Ks) migrate undivided to only one pole of the cell, while the somatic chromosomes (=Ss) first remain in the metaphase plane, and with the arrival of the Ks at the pole, they then separate equally. The evolutionarily conserved GTPase Ran plays a crucial role in many cellular processes. This includes the regulation of microtubule nucleation and stabilisation at kinetochores and of spindle assembly during mitosis, which is promoted by a RanGTP concentration gradient that forms around the mitotic chromosomes (Kalab et al. in Science 295:2452-2456, 2002, Nature 440:697-701, 2006). In the present study, a strong accumulation of Ran was detected by immunofluorescence at the kinetochores of the Ss in normal gonial and differential gonial mitoses of males of A. lucidus. In contrast, no Ran accumulation was observed at the kinetochores of the Ss in the metaphases of brain ganglia mitoses or of aberrant spermatocytes or in metaphases I and II of spermatocyte meiotic divisions. Likewise, there was no accumulation at the kinetochores of Drosophila melanogaster mitotic chromosomes from larval brains. The specific accumulation of Ran at the kinetochores of the Ss in differential gonial mitoses of A. lucidus strongly suggests that Ran is involved in a mechanism acting in this exceptional mitosis, which retains the Ss at the metaphase plane and prevents a premature separation and unequal segregation of the Ss during monopolar migration of the Ks.

ARHGEF17 is an essential spindle assembly checkpoint factor that targets Mps1 to kinetochores

PubMed Central

Isokane, Mayumi; Walter, Thomas; Mahen, Robert; Nijmeijer, Bianca; Hériché, Jean-Karim; Miura, Kota; Maffini, Stefano; Ivanov, Miroslav Penchev; Kitajima, Tomoya S.; Peters, Jan-Michael

2016-01-01

To prevent genome instability, mitotic exit is delayed until all chromosomes are properly attached to the mitotic spindle by the spindle assembly checkpoint (SAC). In this study, we characterized the function of ARHGEF17, identified in a genome-wide RNA interference screen for human mitosis genes. Through a series of quantitative imaging, biochemical, and biophysical experiments, we showed that ARHGEF17 is essential for SAC activity, because it is the major targeting factor that controls localization of the checkpoint kinase Mps1 to the kinetochore. This mitotic function is mediated by direct interaction of the central domain of ARHGEF17 with Mps1, which is autoregulated by the activity of Mps1 kinase, for which ARHGEF17 is a substrate. This mitosis-specific role is independent of ARHGEF17’s RhoGEF activity in interphase. Our study thus assigns a new mitotic function to ARHGEF17 and reveals the molecular mechanism for a key step in SAC establishment. PMID:26953350

ARHGEF17 is an essential spindle assembly checkpoint factor that targets Mps1 to kinetochores.

PubMed

Isokane, Mayumi; Walter, Thomas; Mahen, Robert; Nijmeijer, Bianca; Hériché, Jean-Karim; Miura, Kota; Maffini, Stefano; Ivanov, Miroslav Penchev; Kitajima, Tomoya S; Peters, Jan-Michael; Ellenberg, Jan

2016-03-14

To prevent genome instability, mitotic exit is delayed until all chromosomes are properly attached to the mitotic spindle by the spindle assembly checkpoint (SAC). In this study, we characterized the function of ARHGEF17, identified in a genome-wide RNA interference screen for human mitosis genes. Through a series of quantitative imaging, biochemical, and biophysical experiments, we showed that ARHGEF17 is essential for SAC activity, because it is the major targeting factor that controls localization of the checkpoint kinase Mps1 to the kinetochore. This mitotic function is mediated by direct interaction of the central domain of ARHGEF17 with Mps1, which is autoregulated by the activity of Mps1 kinase, for which ARHGEF17 is a substrate. This mitosis-specific role is independent of ARHGEF17's RhoGEF activity in interphase. Our study thus assigns a new mitotic function to ARHGEF17 and reveals the molecular mechanism for a key step in SAC establishment. © 2016 Isokane et al.

Chromokinesin Kid and kinetochore kinesin CENP-E differentially support chromosome congression without end-on attachment to microtubules.

PubMed

Iemura, Kenji; Tanaka, Kozo

2015-03-06

Chromosome congression is the alignment of chromosomes at the spindle equator, and is a prerequisite for faithful chromosome segregation. Recent data suggest that before kinetochores attach to the end of microtubules (end-on attachment), chromosomes can move along microtubules towards the spindle equator through attachment of kinetochores to the lateral surface of microtubules (lateral attachment). Here we address this mechanism, focusing on the contribution of two mitotic motors, Kid and CENP-E. In cells depleted of Hec1, which is essential for end-on attachment, chromosomes show partial and transient congression. This transient congression is further perturbed by co-depletion of Kid, suggesting its role in chromosome congression. In comparison, CENP-E suppresses chromosome congression, probably by tethering kinetochores to short, unstable microtubules, and works in congression only when microtubules are stabilized. Our results may reflect the differential contributions of Kid and CENP-E in chromosome congression in physiological conditions where stabilized microtubules are becoming increased.

Multivalency of NDC80 in the outer kinetochore is essential to track shortening microtubules and generate forces

PubMed Central

2018-01-01

Presence of multiple copies of the microtubule-binding NDC80 complex is an evolutionary conserved feature of kinetochores, points of attachment of chromosomes to spindle microtubules. This may enable multivalent attachments to microtubules, with implications that remain unexplored. Using recombinant human kinetochore components, we show that while single NDC80 complexes do not track depolymerizing microtubules, reconstituted particles containing the NDC80 receptor CENP-T bound to three or more NDC80 complexes do so effectively, as expected for a kinetochore force coupler. To study multivalency systematically, we engineered modules allowing incremental addition of NDC80 complexes. The modules’ residence time on microtubules increased exponentially with the number of NDC80 complexes. Modules with two or more complexes tracked depolymerizing microtubules with increasing efficiencies, and stalled and rescued microtubule depolymerization in a force-dependent manner when conjugated to cargo. Our observations indicate that NDC80, rather than through biased diffusion, tracks depolymerizing microtubules by harnessing force generated during microtubule disassembly. PMID:29629870

Merotelic kinetochore attachment in oocyte meiosis II causes sister chromatids segregation errors in aged mice.

PubMed

Cheng, Jin-Mei; Li, Jian; Tang, Ji-Xin; Hao, Xiao-Xia; Wang, Zhi-Peng; Sun, Tie-Cheng; Wang, Xiu-Xia; Zhang, Yan; Chen, Su-Ren; Liu, Yi-Xun

2017-08-03

Mammalian oocyte chromosomes undergo 2 meiotic divisions to generate haploid gametes. The frequency of chromosome segregation errors during meiosis I increase with age. However, little attention has been paid to the question of how aging affects sister chromatid segregation during oocyte meiosis II. More importantly, how aneuploid metaphase II (MII) oocytes from aged mice evade the spindle assembly checkpoint (SAC) mechanism to complete later meiosis II to form aneuploid embryos remains unknown. Here, we report that MII oocytes from naturally aged mice exhibited substantial errors in chromosome arrangement and configuration compared with young MII oocytes. Interestingly, these errors in aged oocytes had no impact on anaphase II onset and completion as well as 2-cell formation after parthenogenetic activation. Further study found that merotelic kinetochore attachment occurred more frequently and could stabilize the kinetochore-microtubule interaction to ensure SAC inactivation and anaphase II onset in aged MII oocytes. This orientation could persist largely during anaphase II in aged oocytes, leading to severe chromosome lagging and trailing as well as delay of anaphase II completion. Therefore, merotelic kinetochore attachment in oocyte meiosis II exacerbates age-related genetic instability and is a key source of age-dependent embryo aneuploidy and dysplasia.

Centromere Protein (CENP)-W Interacts with Heterogeneous Nuclear Ribonucleoprotein (hnRNP) U and May Contribute to Kinetochore-Microtubule Attachment in Mitotic Cells

PubMed Central

Chun, Younghwa; Kim, Raehyung; Lee, Soojin

2016-01-01

Background Recent studies have shown that heterogeneous nuclear ribonucleoprotein U (hnRNP U), a component of the hnRNP complex, contributes to stabilize the kinetochore-microtubule interaction during mitosis. CENP-W was identified as an inner centromere component that plays crucial roles in the formation of a functional kinetochore complex. Results We report that hnRNP U interacts with CENP-W, and the interaction between hnRNP U and CENP-W mutually increased each other’s protein stability by inhibiting the proteasome-mediated degradation. Further, their co-localization was observed chiefly in the nuclear matrix region and at the microtubule-kinetochore interface during interphase and mitosis, respectively. Both microtubule-stabilizing and microtubule-destabilizing agents significantly decreased the protein stability of CENP-W. Furthermore, loss of microtubules and defects in microtubule organization were observed in CENP-W-depleted cells. Conclusion Our data imply that CENP-W plays an important role in the attachment and interaction between microtubules and kinetochore during mitosis. PMID:26881882

Kinetochore localized Mad2 and Cdc20 is itself insufficient for triggering the mitotic checkpoint when Mps1 is low in Drosophila melanogaster neuroblasts.

PubMed

Herriott, Ashleigh; Sweeney, Michele; Whitaker, Michael; Taggart, Michael; Huang, Jun-Yong

2012-12-15

The relationships between the kinetochore and checkpoint control remain unresolved. Here, we report the characterization of the in vivo behavior of Cdc20 and Mad2 and the relevant spindle assembly checkpoint (SAC) functions in the neuroblasts of a Drosophila Mps1 weak allele (ald (B4-2) ). ald (B4-2) third instar larvae brain samples contain only around 16% endogenous Mps1 protein, and the SAC function is abolished. However, this does not lead to rapid anaphase onset and mitotic exit, in contrast to the loss of Mad2 alone in a mad2 (EY) mutant. The level of GFP-Cdc20 recruitment to the kinetochore is unaffected in ald (B4-2) neuroblasts, while the level of GFP-Mad2 is reduced to just about 20%. Cdc20 and Mad2 display only monophasic exponential kinetics at the kinetochores. The ald (B4-2) heterozygotes expressed approximately 65% of normal Mps1 protein levels, and this is enough to restore the SAC function. The kinetochore recruitment of GFP-Mad2 in response to SAC activation increases by around 80% in heterozygotes, compared with just about 20% in ald (B4-2) mutant. This suggests a correlation between Mps1 levels and Mad2 kinetochore localization and perhaps the existence of a threshold level at which Mps1 is fully functional. The failure to arrest the mitotic progression in ald (B4-2) neuroblasts in response to colchicine treatment suggests that when Mps1 levels are low, approximately 20% of normal GFP-Mad2, alongside normal levels of GFP-Cdc20 kinetochore recruitments, is insufficient for triggering SAC signal propagation.

PP2A regulates kinetochore-microtubule attachment during meiosis I in oocyte.

PubMed

Tang, An; Shi, Peiliang; Song, Anying; Zou, Dayuan; Zhou, Yue; Gu, Pengyu; Huang, Zan; Wang, Qinghua; Lin, Zhaoyu; Gao, Xiang

2016-06-02

Studies using in vitro cultured oocytes have indicated that the protein phosphatase 2A (PP2A), a major serine/threonine protein phosphatase, participates in multiple steps of meiosis. Details of oocyte maturation regulation by PP2A remain unclear and an in vivo model can provide more convincing information. Here, we inactivated PP2A by mutating genes encoding for its catalytic subunits (PP2Acs) in mouse oocytes. We found that eliminating both PP2Acs caused female infertility. Oocytes lacking PP2Acs failed to complete 1(st) meiotic division due to chromosome misalignment and abnormal spindle assembly. In mitosis, PP2A counteracts Aurora kinase B/C (AurkB/C) to facilitate correct kinetochore-microtubule (KT-MT) attachment. In meiosis I in oocyte, we found that PP2Ac deficiency destabilized KT-MT attachments. Chemical inhibition of AurkB/C in PP2Ac-null oocytes partly restored the formation of lateral/merotelic KT-MT attachments but not correct KT-MT attachments. Taken together, our findings demonstrate that PP2Acs are essential for chromosome alignments and regulate the formation of correct KT-MT attachments in meiosis I in oocytes.

Mitosis in Barbulanympha. I. Spindle structure, formation, and kinetochore engagement

PubMed Central

1978-01-01

Successful culture of the obligatorily anaerobic symbionts residing in the hindgut of the wood-eating cockroach Cryptocercus punctulatus now permits continuous observation of mitosis in individual Barbulanympha cells. In Part I of this two-part paper, we report methods for culture of the protozoa, preparation of microscope slide cultures in which Barbulanympha survived and divided for up to 3 days, and an optical arrangement which permits observation and through-focus photographic recording of dividing cells, sequentially in differential interference contrast and rectified polarized light microscopy. We describe the following prophase events and structures: development of the astral rays and large extranuclear central spindle from the tips of the elongate-centrioles; the fine structure of spindle fibers and astral rays which were deduced in vivo from polarized light microscopy and seen as a particular array of microtubules in thin-section electron micrographs; formation of chromosomal spindle fibers by dynamic engagement of astral rays to the kinetochores embedded in the persistent nuclear envelope; and repetitive shortening of chromosomal spindle fibers which appear to hoist the nucleus to the spindle surface, cyclically jostle the kinetochores within the nuclear envelope, and churn the prophase chromosomes. The observations described here and in Part II have implications both for the evolution of mitosis and for understanding the mitotic process generally. PMID:681451

MiCroKit 3.0: an integrated database of midbody, centrosome and kinetochore.

PubMed

Ren, Jian; Liu, Zexian; Gao, Xinjiao; Jin, Changjiang; Ye, Mingliang; Zou, Hanfa; Wen, Longping; Zhang, Zhaolei; Xue, Yu; Yao, Xuebiao

2010-01-01

During cell division/mitosis, a specific subset of proteins is spatially and temporally assembled into protein super complexes in three distinct regions, i.e. centrosome/spindle pole, kinetochore/centromere and midbody/cleavage furrow/phragmoplast/bud neck, and modulates cell division process faithfully. Although many experimental efforts have been carried out to investigate the characteristics of these proteins, no integrated database was available. Here, we present the MiCroKit database (http://microkit.biocuckoo.org) of proteins that localize in midbody, centrosome and/or kinetochore. We collected into the MiCroKit database experimentally verified microkit proteins from the scientific literature that have unambiguous supportive evidence for subcellular localization under fluorescent microscope. The current version of MiCroKit 3.0 provides detailed information for 1489 microkit proteins from seven model organisms, including Saccharomyces cerevisiae, Schizasaccharomyces pombe, Caenorhabditis elegans, Drosophila melanogaster, Xenopus laevis, Mus musculus and Homo sapiens. Moreover, the orthologous information was provided for these microkit proteins, and could be a useful resource for further experimental identification. The online service of MiCroKit database was implemented in PHP + MySQL + JavaScript, while the local packages were developed in JAVA 1.5 (J2SE 5.0).

The kinetochore proteins CENP-E and CENP-F directly and specifically interact with distinct BUB mitotic checkpoint Ser/Thr kinases.

PubMed

Ciossani, Giuseppe; Overlack, Katharina; Petrovic, Arsen; Huis In 't Veld, Pim J; Koerner, Carolin; Wohlgemuth, Sabine; Maffini, Stefano; Musacchio, Andrea

2018-05-10

The segregation of chromosomes during cell division relies on the function of the kinetochores, protein complexes that physically connect chromosomes with microtubules of the spindle. The metazoan proteins, centromere protein E (CENP-E) and CENP-F, are components of a fibrous layer of mitotic kinetochores named the corona. Several of their features suggest that CENP-E and CENP-F are paralogs: they are very large (comprising approximately 2700 and 3200 residues, respectively), contain abundant predicted coiled-coil structures, are C-terminally prenylated, and are endowed with microtubule-binding sites at their termini. Moreover, CENP-E contains an ATP-hydrolyzing motor domain that promotes microtubule plus end-directed motion. Here, we show that both CENP-E and CENP-F are recruited to mitotic kinetochores independently of the main corona constituent, the Rod-Zwilch-ZW10 (RZZ) complex. We identified specific interactions of CENP-F and CENP-E with budding uninhibited by benzimidazole 1 (BUB1) and BUB1-related (BUBR1) mitotic checkpoint Ser/Thr kinases, respectively, paralogous proteins involved in mitotic checkpoint control and chromosome alignment. Whereas BUBR1 was dispensable for kinetochore localization of CENP-E, BUB1 was stringently required for CENP-F localization. Through biochemical reconstitution, we demonstrated that the CENP-E-BUBR1 and CENP-F-BUB1 interactions are direct and require similar determinants, a dimeric coiled-coil in CENP-E or CENP-F and a kinase domain in BUBR1 or BUB1. Our findings are consistent with the existence of structurally similar BUB1-CENP-F and BUBR1-CENP-E complexes, supporting the notion that CENP-E and CENP-F are evolutionarily related. Published under license by The American Society for Biochemistry and Molecular Biology, Inc.

MiCroKit 3.0: an integrated database of midbody, centrosome and kinetochore

PubMed Central

Liu, Zexian; Gao, Xinjiao; Jin, Changjiang; Ye, Mingliang; Zou, Hanfa; Wen, Longping; Zhang, Zhaolei; Xue, Yu; Yao, Xuebiao

2010-01-01

During cell division/mitosis, a specific subset of proteins is spatially and temporally assembled into protein super complexes in three distinct regions, i.e. centrosome/spindle pole, kinetochore/centromere and midbody/cleavage furrow/phragmoplast/bud neck, and modulates cell division process faithfully. Although many experimental efforts have been carried out to investigate the characteristics of these proteins, no integrated database was available. Here, we present the MiCroKit database (http://microkit.biocuckoo.org) of proteins that localize in midbody, centrosome and/or kinetochore. We collected into the MiCroKit database experimentally verified microkit proteins from the scientific literature that have unambiguous supportive evidence for subcellular localization under fluorescent microscope. The current version of MiCroKit 3.0 provides detailed information for 1489 microkit proteins from seven model organisms, including Saccharomyces cerevisiae, Schizasaccharomyces pombe, Caenorhabditis elegans, Drosophila melanogaster, Xenopus laevis, Mus musculus and Homo sapiens. Moreover, the orthologous information was provided for these microkit proteins, and could be a useful resource for further experimental identification. The online service of MiCroKit database was implemented in PHP + MySQL + JavaScript, while the local packages were developed in JAVA 1.5 (J2SE 5.0). PMID:19783819

Coupling unbiased mutagenesis to high-throughput DNA sequencing uncovers functional domains in the Ndc80 kinetochore protein of Saccharomyces cerevisiae.

PubMed

Tien, Jerry F; Fong, Kimberly K; Umbreit, Neil T; Payen, Celia; Zelter, Alex; Asbury, Charles L; Dunham, Maitreya J; Davis, Trisha N

2013-09-01

During mitosis, kinetochores physically link chromosomes to the dynamic ends of spindle microtubules. This linkage depends on the Ndc80 complex, a conserved and essential microtubule-binding component of the kinetochore. As a member of the complex, the Ndc80 protein forms microtubule attachments through a calponin homology domain. Ndc80 is also required for recruiting other components to the kinetochore and responding to mitotic regulatory signals. While the calponin homology domain has been the focus of biochemical and structural characterization, the function of the remainder of Ndc80 is poorly understood. Here, we utilized a new approach that couples high-throughput sequencing to a saturating linker-scanning mutagenesis screen in Saccharomyces cerevisiae. We identified domains in previously uncharacterized regions of Ndc80 that are essential for its function in vivo. We show that a helical hairpin adjacent to the calponin homology domain influences microtubule binding by the complex. Furthermore, a mutation in this hairpin abolishes the ability of the Dam1 complex to strengthen microtubule attachments made by the Ndc80 complex. Finally, we defined a C-terminal segment of Ndc80 required for tetramerization of the Ndc80 complex in vivo. This unbiased mutagenesis approach can be generally applied to genes in S. cerevisiae to identify functional properties and domains.

Kif18A and chromokinesins confine centromere movements via microtubule growth suppression and spatial control of kinetochore tension

PubMed Central

Stumpff, Jason; Wagenbach, Michael; Franck, Andrew; Asbury, Charles L.; Wordeman, Linda

2012-01-01

Summary Alignment of chromosomes at the metaphase plate is a signature of cell division in metazoan cells, yet the mechanisms controlling this process remain ambiguous. Here we use a combination of quantitative live cell imaging and reconstituted dynamic microtubule assays to investigate the molecular control of mitotic centromere movements. We establish that Kif18A (kinesin-8) attenuates centromere movement by directly promoting microtubule pausing in a concentration-dependent manner. This activity provides the dominant mechanism for restricting centromere movement to the spindle midzone. Furthermore, polar ejection forces spatially confine chromosomes via position-dependent regulation of kinetochore tension and centromere switch rates. We demonstrate that polar ejection forces are antagonistically modulated by chromokinesins. These pushing forces depend on Kid (kinesin-10) activity and are antagonized by Kif4A (kinesin-4), which functions to directly suppress microtubule growth. These data support a model in which Kif18A and polar ejection forces synergistically promote centromere alignment via spatial control of kinetochore-microtubule dynamics. PMID:22595673

A minimal model for kinetochore-microtubule dynamics

NASA Astrophysics Data System (ADS)

Liu, Andrea

2014-03-01

During mitosis, chromosome pairs align at the center of a bipolar microtubule (MT) spindle and oscillate as MTs attaching them to the cell poles polymerize and depolymerize. The cell fixes misaligned pairs by a tension-sensing mechanism. Pairs later separate as shrinking MTs pull each chromosome toward its respective cell pole. We present a minimal model for these processes based on properties of MT kinetics. We apply the measured tension-dependence of single MT kinetics to a stochastic many MT model, which we solve numerically and with master equations. We find that the force-velocity curve for the single chromosome system is bistable and hysteretic. Above some threshold load, tension fluctuations induce MTs to spontaneously switch from a pulling state into a growing, pushing state. To recover pulling from the pushing state, the load must be reduced far below the threshold. This leads to oscillations in the two-chromosome system. Our minimal model quantitatively captures several aspects of kinetochore dynamics observed experimentally. This work was supported by NSF-DMR-1104637.

Expression of Spindle and Kinetochore-Associated Protein 1 Is Associated with Poor Prognosis in Papillary Thyroid Carcinoma

PubMed Central

Dong, Chao; Wang, Xiao-li; Ma, Bin-lin

2015-01-01

Aim. Spindle and kinetochore-associated protein 1 (SKA1) is one subtype of SKA, whose protein can make spindle microtubules attach steadily to the kinetochore in the middle of mitosis. At present, there are fewer researches on the relationship between SKA1 expression and tumor development. Methods. In this study, immunohistochemical analysis was used to determine the expression of SKA1 in papillary thyroid carcinoma (PTC) and adjacent tissues. We used quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot analysis to further verify the results. Results. We found that SKA1 expression was significantly higher in PTC tissues than normal adjacent tissues (P < 0.05). There existed a significant correlation among a higher SKA1 expression, including lymphoid node (P = 0.005), clinical stage (P = 0.015), and extrathyroid invasion (P = 0.004). Survival analysis showed high SKA1 expression in PTC patients more likely to relapse after surgery. Conclusion. High SKA1 expression is predictive of poor prognosis of PTC, implying that SKA1 may be a promising new target for targeted therapies for PTC. PMID:26063960

CENP-C and CENP-I are key connecting factors for kinetochore and CENP-A assembly

PubMed Central

Shono, Nobuaki; Ohzeki, Jun-ichirou; Otake, Koichiro; Martins, Nuno M. C.; Nagase, Takahiro; Kimura, Hiroshi; Larionov, Vladimir; Earnshaw, William C.; Masumoto, Hiroshi

2015-01-01

ABSTRACT Although it is generally accepted that chromatin containing the histone H3 variant CENP-A is an epigenetic mark maintaining centromere identity, the pathways leading to the formation and maintenance of centromere chromatin remain unclear. We previously generated human artificial chromosomes (HACs) whose centromeres contain a synthetic alpha-satellite (alphoid) DNA array containing the tetracycline operator (alphoidtetO). We also obtained cell lines bearing the alphoidtetO array at ectopic integration sites on chromosomal arms. Here, we have examined the regulation of CENP-A assembly at centromeres as well as de novo assembly on the ectopic arrays by tethering tetracycline repressor (tetR) fusions of substantial centromeric factors and chromatin modifiers. This analysis revealed four classes of factors that influence CENP-A assembly. Interestingly, many kinetochore structural components induced de novo CENP-A assembly at the ectopic site. We showed that these components work by recruiting CENP-C and subsequently recruiting M18BP1. Furthermore, we found that CENP-I can also recruit M18BP1 and, as a consequence, enhances M18BP1 assembly on centromeres in the downstream of CENP-C. Thus, we suggest that CENP-C and CENP-I are key factors connecting kinetochore to CENP-A assembly. PMID:26527398

Conserved tetramer junction in the kinetochore Ndc80 complex

PubMed Central

Valverde, Roberto; Ingram, Jessica; Harrison, Stephen C.

2016-01-01

Summary The heterotetrameric Ndc80 complex establishes connectivity along the principal longitudinal axis of a kinetochore. Its two heterodimeric subcomplexes, each with a globular end and a coiled-coil shaft, connect end-to-end to create a ∼600 Å long rod spanning the gap from centromere-proximal structures to spindle microtubules. Neither subcomplex has a known function on its own, but the heterotetrameric organization and the characteristics of the junction are conserved from yeast to man. We have determined crystal structures of two shortened (“dwarf”) Ndc80 complexes that contain the full tetramer junction and both globular ends. The junction connects two α-helical coiled coils through regions of four-chain and three-chain overlap. The complexity of its structure depends on interactions among conserved amino-acid residues, suggesting a binding site for additional cellular factor(s) not yet identified. PMID:27851957

The Nuclear Transport Factor Kap121 Is Required for Stability of the Dam1 Complex and Mitotic Kinetochore Bi-orientation.

PubMed

Cairo, Lucas V; Wozniak, Richard W

2016-03-15

The karyopherin (Kap) family of nuclear transport factors facilitates macromolecular transport through nuclear pore complexes (NPCs). The binding of Kaps to their cargos can also regulate, both temporally and spatially, the interactions of the cargo protein with interacting partners. Here, we show that the essential yeast Kap, Kap121, binds Dam1 and Duo1, components of the microtubule (MT)-associated Dam1 complex required for linking dynamic MT ends with kinetochores (KTs). Like mutations in the Dam1 complex, loss of Kap121 function compromises the formation of normal KT-MT attachments during mitosis. We show that the stability of the Dam1 complex in vivo is dependent on its association with Kap121. Furthermore, we show that the Kap121/Duo1 complex is maintained in the presence of RanGTP but Kap121 is released by the cooperative actions of RanGTP and tubulin. We propose that Kap121 stabilizes the Dam1 complex and participates in escorting it to spindle MTs. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

Contributions of microtubule rotation and dynamic instability to kinetochore capture

NASA Astrophysics Data System (ADS)

Sweezy-Schindler, Oliver; Edelmaier, Christopher; Blackwell, Robert; Glaser, Matt; Betterton, Meredith

2014-03-01

The capture of lost kinetochores (KCs) by microtubules (MTs) is a crucial part of prometaphase during mitosis. Microtubule dynamic instability has been considered the primary mechanism of KC capture, but recent work discovered that lateral KC attachment to pivoting MTs enabled rapid capture even with significantly reduced MT dynamics. We aim to understand the relative contributions of MT rotational diffusion and dynamic instability to KC capture, as well as KC capture through end-on and/or lateral attachment. Our model consists of rigid MTs and a spherical KC, which are allowed to diffuse inside a spherical nuclear envelope consistent with the geometry of fission yeast. For simplicity, we include a single spindle pole body, which is anchored to the nuclear membrane, and its associated polar MTs. Brownian dynamics treats the diffusion of the MTs and KC and kinetic Monte Carlo models stochastic processes such as dynamic instability. NSF 1546021.

The CENP-T/-W complex is a binding partner of the histone chaperone FACT

PubMed Central

Prendergast, Lisa; Müller, Sebastian; Liu, Yiwei; Huang, Hongda; Dingli, Florent; Loew, Damarys; Vassias, Isabelle; Patel, Dinshaw J.; Sullivan, Kevin F.; Almouzni, Geneviève

2016-01-01

The CENP-T/-W histone fold complex, as an integral part of the inner kinetochore, is essential for building a proper kinetochore at the centromere in order to direct chromosome segregation during mitosis. Notably, CENP-T/-W is not inherited at centromeres, and new deposition is absolutely required at each cell cycle for kinetochore function. However, the mechanisms underlying this new deposition of CENP-T/-W at centromeres are unclear. Here, we found that CENP-T deposition at centromeres is uncoupled from DNA synthesis. We identified Spt16 and SSRP1, subunits of the H2A–H2B histone chaperone facilitates chromatin transcription (FACT), as CENP-W binding partners through a proteomic screen. We found that the C-terminal region of Spt16 binds specifically to the histone fold region of CENP-T/-W. Furthermore, depletion of Spt16 impairs CENP-T and CENP-W deposition at endogenous centromeres, and site-directed targeting of Spt16 alone is sufficient to ensure local de novo CENP-T accumulation. We propose a model in which the FACT chaperone stabilizes the soluble CENP-T/-W complex in the cell and promotes dynamics of exchange, enabling CENP-T/-W deposition at centromeres. PMID:27284163

The CENP-T/-W complex is a binding partner of the histone chaperone FACT.

PubMed

Prendergast, Lisa; Müller, Sebastian; Liu, Yiwei; Huang, Hongda; Dingli, Florent; Loew, Damarys; Vassias, Isabelle; Patel, Dinshaw J; Sullivan, Kevin F; Almouzni, Geneviève

2016-06-01

The CENP-T/-W histone fold complex, as an integral part of the inner kinetochore, is essential for building a proper kinetochore at the centromere in order to direct chromosome segregation during mitosis. Notably, CENP-T/-W is not inherited at centromeres, and new deposition is absolutely required at each cell cycle for kinetochore function. However, the mechanisms underlying this new deposition of CENP-T/-W at centromeres are unclear. Here, we found that CENP-T deposition at centromeres is uncoupled from DNA synthesis. We identified Spt16 and SSRP1, subunits of the H2A-H2B histone chaperone facilitates chromatin transcription (FACT), as CENP-W binding partners through a proteomic screen. We found that the C-terminal region of Spt16 binds specifically to the histone fold region of CENP-T/-W. Furthermore, depletion of Spt16 impairs CENP-T and CENP-W deposition at endogenous centromeres, and site-directed targeting of Spt16 alone is sufficient to ensure local de novo CENP-T accumulation. We propose a model in which the FACT chaperone stabilizes the soluble CENP-T/-W complex in the cell and promotes dynamics of exchange, enabling CENP-T/-W deposition at centromeres. © 2016 Prendergast et al.; Published by Cold Spring Harbor Laboratory Press.

On the robustness of SAC silencing in closed mitosis

NASA Astrophysics Data System (ADS)

Ruth, Donovan; Liu, Jian

Mitosis equally partitions sister chromatids to two daughter cells. This is achieved by properly attaching these chromatids via their kinetochores to microtubules that emanate from the spindle poles. Once the last kinetochore is properly attached, the spindle microtubules pull the sister chromatids apart. Due to the dynamic nature of microtubules, however, kinetochore-microtubule attachment often goes wrong. When this erroneous attachment occurs, it locally activates an ensemble of proteins, called the spindle assembly checkpoint proteins (SAC), which halts the mitotic progression until all the kinetochores are properly attached by spindle microtubules. The timing of SAC silencing thus determines the fidelity of chromosome segregation. We previously established a spatiotemporal model that addresses the robustness of SAC silencing in open mitosis for the first time. Here, we focus on closed mitosis by examining yeast mitosis as a model system. Though much experimental work has been done to study the SAC in cells undergoing closed mitosis, the processes responsible are not well understood. We leverage and extend our previous model to study SAC silencing mechanism in closed mitosis. We show that a robust signal of the SAC protein accumulation at the spindle pole body can be achieved. This signal is a nonlinear increasing function of number of kinetochore-microtubule attachments, and can thus serve as a robust trigger to time the SAC silencing. Together, our mechanism provides a unified framework across species that ensures robust SAC silencing and fidelity of chromosome segregation in mitosis. Intramural research program in NHLBI at NIH.

Mechanism for G2 phase-specific nuclear export of the kinetochore protein CENP-F.

PubMed

Loftus, Kyle M; Cui, Heying; Coutavas, Elias; King, David S; Ceravolo, Amanda; Pereiras, Dylan; Solmaz, Sozanne R

2017-08-03

Centromere protein F (CENP-F) is a component of the kinetochore and a regulator of cell cycle progression. CENP-F recruits the dynein transport machinery and orchestrates several cell cycle-specific transport events, including transport of the nucleus, mitochondria and chromosomes. A key regulatory step for several of these functions is likely the G2 phase-specific export of CENP-F from the nucleus to the cytosol, where the cytoplasmic dynein transport machinery resides; however, the molecular mechanism of this process is elusive. Here, we have identified 3 phosphorylation sites within the bipartite classical nuclear localization signal (cNLS) of CENP-F. These sites are specific for cyclin-dependent kinase 1 (Cdk1), which is active in G2 phase. Phosphomimetic mutations of these residues strongly diminish the interaction of the CENP-F cNLS with its nuclear transport receptor karyopherin α. These mutations also diminish nuclear localization of the CENP-F cNLS in cells. Notably, the cNLS is phosphorylated in the -1 position, which is important to orient the adjacent major motif for binding into its pocket on karyopherin α. We propose that localization of CENP-F is regulated by a cNLS, and a nuclear export pathway, resulting in nuclear localization during most of interphase. In G2 phase, the cNLS is weakened by phosphorylation through Cdk1, likely resulting in nuclear export of CENP-F via the still active nuclear export pathway. Once CENP-F resides in the cytosol, it can engage in pathways that are important for cell cycle progression, kinetochore assembly and the faithful segregation of chromosomes into daughter cells.

The Microtubule-Stabilizing Protein CLASP1 Associates with the Theileria annulata Schizont Surface via Its Kinetochore-Binding Domain

PubMed Central

Huber, Sandra; Theiler, Romina; de Quervain, Daniel; Wiens, Olga; Karangenc, Tulin; Heussler, Volker; Dobbelaere, Dirk

2017-01-01

ABSTRACT Theileria is an apicomplexan parasite whose presence within the cytoplasm of a leukocyte induces cellular transformation and causes uncontrolled proliferation and clonal expansion of the infected cell. The intracellular schizont utilizes the host cell’s own mitotic machinery to ensure its distribution to both daughter cells by associating closely with microtubules (MTs) and incorporating itself within the central spindle. We show that CLASP1, an MT-stabilizing protein that plays important roles in regulating kinetochore-MT attachment and central spindle positioning, is sequestered at the Theileria annulata schizont surface. We used live-cell imaging and immunofluorescence in combination with MT depolymerization assays to demonstrate that CLASP1 binds to the schizont surface in an MT-independent manner throughout the cell cycle and that the recruitment of the related CLASP2 protein to the schizont is MT dependent. By transfecting Theileria-infected cells with a panel of truncation mutants, we found that the kinetochore-binding domain of CLASP1 is necessary and sufficient for parasite localization, revealing that CLASP1 interaction with the parasite occurs independently of EB1. We overexpressed the MT-binding domain of CLASP1 in parasitized cells. This exhibited a dominant negative effect on host MT stability and led to altered parasite size and morphology, emphasizing the importance of proper MT dynamics for Theileria partitioning during host cell division. Using coimmunoprecipitation, we demonstrate that CLASP1 interacts, directly or indirectly, with the schizont membrane protein p104, and we describe for the first time TA03615, a Theileria protein which localizes to the parasite surface, where it has the potential to participate in parasite-host interactions. IMPORTANCE T. annulata, the only eukaryote known to be capable of transforming another eukaryote, is a widespread parasite of veterinary importance that puts 250 million cattle at risk worldwide

CENP-A regulates chromosome segregation during the first meiosis of mouse oocytes.

PubMed

Li, Li; Qi, Shu-Tao; Sun, Qing-Yuan; Chen, Shi-Ling

2017-06-01

Proper chromosome separation in both mitosis and meiosis depends on the correct connection between kinetochores of chromosomes and spindle microtubules. Kinetochore dysfunction can lead to unequal distribution of chromosomes during cell division and result in aneuploidy, thus kinetochores are critical for faithful segregation of chromosomes. Centromere protein A (CENP-A) is an important component of the inner kinetochore plate. Multiple studies in mitosis have found that deficiencies in CENP-A could result in structural and functional changes of kinetochores, leading to abnormal chromosome segregation, aneuploidy and apoptosis in cells. Here we report the expression and function of CENP-A during mouse oocyte meiosis. Our study found that microinjection of CENP-A blocking antibody resulted in errors of homologous chromosome segregation and caused aneuploidy in eggs. Thus, our findings provide evidence that CENP-A is critical for the faithful chromosome segregation during mammalian oocyte meiosis.

Direct kinetochore–spindle pole connections are not required for chromosome segregation

PubMed Central

Sikirzhytski, Vitali; Magidson, Valentin; Steinman, Jonathan B.; He, Jie; Le Berre, Maël; Tikhonenko, Irina; Ault, Jeffrey G.; McEwen, Bruce F.; Chen, James K.; Sui, Haixin; Piel, Matthieu; Kapoor, Tarun M.

2014-01-01

Segregation of genetic material occurs when chromosomes move to opposite spindle poles during mitosis. This movement depends on K-fibers, specialized microtubule (MT) bundles attached to the chromosomes′ kinetochores. A long-standing assumption is that continuous K-fibers connect every kinetochore to a spindle pole and the force for chromosome movement is produced at the kinetochore and coupled with MT depolymerization. However, we found that chromosomes still maintained their position at the spindle equator during metaphase and segregated properly during anaphase when one of their K-fibers was severed near the kinetochore with a laser microbeam. We also found that, in normal fully assembled spindles, K-fibers of some chromosomes did not extend to the spindle pole. These K-fibers connected to adjacent K-fibers and/or nonkinetochore MTs. Poleward movement of chromosomes with short K-fibers was uncoupled from MT depolymerization at the kinetochore. Instead, these chromosomes moved by dynein-mediated transport of the entire K-fiber/kinetochore assembly. Thus, at least two distinct parallel mechanisms drive chromosome segregation in mammalian cells. PMID:25023516

Drosophila Polo regulates the spindle assembly checkpoint through Mps1-dependent BubR1 phosphorylation.

PubMed

Conde, Carlos; Osswald, Mariana; Barbosa, João; Moutinho-Santos, Tatiana; Pinheiro, Diana; Guimarães, Sofia; Matos, Irina; Maiato, Helder; Sunkel, Claudio E

2013-06-12

Maintenance of genomic stability during eukaryotic cell division relies on the spindle assembly checkpoint (SAC) that prevents mitotic exit until all chromosomes are properly attached to the spindle. Polo is a mitotic kinase proposed to be involved in SAC function, but its role has remained elusive. We demonstrate that Polo and Aurora B functional interdependency comprises a positive feedback loop that promotes Mps1 kinetochore localization and activity. Expression of constitutively active Polo restores normal Mps1 kinetochore levels even after Aurora B inhibition, highlighting a role for Polo in Mps1 recruitment to unattached kinetochores downstream of Aurora B. We also show that Mps1 kinetochore localization is required for BubR1 hyperphosphorylation and formation of the 3F3/2 phosphoepitope. This is essential to allow recruitment of Cdc20 to unattached kinetochores and the assembly of anaphase-promoting complex/cyclosome-inhibitory complexes to levels that ensure long-term SAC activity. We propose a model in which Polo controls Mps1-dependent BubR1 phosphorylation to promote Cdc20 kinetochore recruitment and sustained SAC function.

Drosophila Polo regulates the spindle assembly checkpoint through Mps1-dependent BubR1 phosphorylation

PubMed Central

Conde, Carlos; Osswald, Mariana; Barbosa, João; Moutinho-Santos, Tatiana; Pinheiro, Diana; Guimarães, Sofia; Matos, Irina; Maiato, Helder; Sunkel, Claudio E

2013-01-01

Maintenance of genomic stability during eukaryotic cell division relies on the spindle assembly checkpoint (SAC) that prevents mitotic exit until all chromosomes are properly attached to the spindle. Polo is a mitotic kinase proposed to be involved in SAC function, but its role has remained elusive. We demonstrate that Polo and Aurora B functional interdependency comprises a positive feedback loop that promotes Mps1 kinetochore localization and activity. Expression of constitutively active Polo restores normal Mps1 kinetochore levels even after Aurora B inhibition, highlighting a role for Polo in Mps1 recruitment to unattached kinetochores downstream of Aurora B. We also show that Mps1 kinetochore localization is required for BubR1 hyperphosphorylation and formation of the 3F3/2 phosphoepitope. This is essential to allow recruitment of Cdc20 to unattached kinetochores and the assembly of anaphase-promoting complex/cyclosome-inhibitory complexes to levels that ensure long-term SAC activity. We propose a model in which Polo controls Mps1-dependent BubR1 phosphorylation to promote Cdc20 kinetochore recruitment and sustained SAC function. PMID:23685359

Development of recombinant adeno-associated virus vectors carrying small interfering RNA (shHec1)-mediated depletion of kinetochore Hec1 protein in tumor cells.

PubMed

Li, L; Yang, L; Scudiero, D A; Miller, S A; Yu, Z-X; Stukenberg, P T; Shoemaker, R H; Kotin, R M

2007-05-01

Transcript depletion using small interfering RNA (siRNA) technology represents a potentially valuable technique for the treatment of cancer. However, delivering therapeutic quantities of siRNA into solid tumors by chemical transfection is not feasible, whereas viral vectors efficiently transduce many human tumor cell lines. Yet producing sufficient quantities of viral vectors that elicit acute and selective cytotoxicity remains a major obstacle for preclinical and clinical trials. Using the invertebrate Spodoptera frugiperda (Sf9) cell line, we were able to produce high titer stocks of cytotoxic recombinant adeno-associated virus (rAAV) that express short hairpin RNA (shRNA) and that efficiently deplete Hec1 (highly expressed in cancer 1), or Kntc2 (kinetochore-associated protein 2), a kinetochore protein directly involved in kinetochore microtubule interactions, chromosome congression and spindle checkpoint signaling. Depletion of Hec1 protein results in persistent spindle checkpoint activation followed by cell death. Because Hec1 expression and activity are only present in mitotic cells, non-dividing cells were not affected by rAAV treatment. On the basis of the results of screening 56 human tumor cell lines with three different serotype vectors, we used a tumor xenograft model to test the effects in vivo. The effects of the shHec1 vector were evident in sectioned and stained tumors. The experiments with rAAV-shRNA vectors demonstrate the utility of producing vectors in invertebrate cells to obtain sufficient concentrations and quantities for solid tumor therapy. This addresses an important requirement for cancer gene therapy, to produce cytotoxic vectors in sufficient quantities and concentrations to enable quantitative transduction and selective killing of solid tumor cells.

Centromeric chromatin and its dynamics in plants.

PubMed

Lermontova, Inna; Sandmann, Michael; Mascher, Martin; Schmit, Anne-Catherine; Chabouté, Marie-Edith

2015-07-01

Centromeres are chromatin structures that are required for proper separation of chromosomes during mitosis and meiosis. The centromere is composed of centromeric DNA, often enriched in satellite repeats, and kinetochore complex proteins. To date, over 100 kinetochore components have been identified in various eukaryotes. Kinetochore assembly begins with incorporation of centromeric histone H3 variant CENH3 into centromeric nucleosomes. Protein components of the kinetochore are either present at centromeres throughout the cell cycle or localize to centromeres transiently, prior to attachment of microtubules to each kinetochore in prometaphase of mitotic cells. This is the case for the spindle assembly checkpoint (SAC) proteins in animal cells. The SAC complex ensures equal separation of chromosomes between daughter nuclei by preventing anaphase onset before metaphase is complete, i.e. the sister kinetochores of all chromosomes are attached to spindle fibers from opposite poles. In this review, we focus on the organization of centromeric DNA and the kinetochore assembly in plants. We summarize recent advances regarding loading of CENH3 into the centromere, and the subcellular localization and protein-protein interactions of Arabidopsis thaliana proteins involved in kinetochore assembly and function. We describe the transcriptional activity of corresponding genes based on in silico analysis of their promoters and cell cycle-dependent expression. Additionally, barley homologs of all selected A. thaliana proteins have been identified in silico, and their sequences and domain structures are presented. © 2015 The Authors The Plant Journal © 2015 John Wiley & Sons Ltd.

Mechanics of kinetochore microtubules and their interactions with chromosomes during cell division

NASA Astrophysics Data System (ADS)

Nazockdast, Ehssan; Fürthauer, Sebastian; Redemann, Stephanie; Baumgart, Johannes; Lindow, Norbert; Kratz, Andrea; Prohaska, Steffen; Müller-Reichert, Thomas; Shelley, Michael

2016-11-01

The accurate segregation of chromosomes, and subsequent cell division, in Eukaryotic cells is achieved by the interactions of an assembly of microtubules (MTs) and motor-proteins, known as the mitotic spindle. We use a combination of our computational platform for simulating cytoskeletal assemblies and our structural data from high-resolution electron tomography of the mitotic spindle, to study the kinetics and mechanics of MTs in the spindle, and their interactions with chromosomes during chromosome segregation in the first cell division in C.elegans embryo. We focus on kinetochore MTs, or KMTs, which have one end attached to a chromosome. KMTs are thought to be a key mechanical component in chromosome segregation. Using exploratory simulations of MT growth, bending, hydrodynamic interactions, and attachment to chromosomes, we propose a mechanical model for KMT-chromosome interactions that reproduces observed KMT length and shape distributions from electron tomography. We find that including detailed hydrodynamic interactions between KMTs is essential for agreement with the experimental observations.

The Ndc80 complex bridges two Dam1 complex rings

PubMed Central

Kim, Jae ook; Zelter, Alex; Umbreit, Neil T; Bollozos, Athena; Riffle, Michael; Johnson, Richard; MacCoss, Michael J; Asbury, Charles L; Davis, Trisha N

2017-01-01

Strong kinetochore-microtubule attachments are essential for faithful segregation of sister chromatids during mitosis. The Dam1 and Ndc80 complexes are the main microtubule binding components of the Saccharomyces cerevisiae kinetochore. Cooperation between these two complexes enhances kinetochore-microtubule coupling and is regulated by Aurora B kinase. We show that the Ndc80 complex can simultaneously bind and bridge across two Dam1 complex rings through a tripartite interaction, each component of which is regulated by Aurora B kinase. Mutations in any one of the Ndc80p interaction regions abrogates the Ndc80 complex’s ability to bind two Dam1 rings in vitro, and results in kinetochore biorientation and microtubule attachment defects in vivo. We also show that an extra-long Ndc80 complex, engineered to space the two Dam1 rings further apart, does not support growth. Taken together, our work suggests that each kinetochore in vivo contains two Dam1 rings and that proper spacing between the rings is vital. DOI: http://dx.doi.org/10.7554/eLife.21069.001 PMID:28191870

Multipolar Spindle Pole Coalescence Is a Major Source of Kinetochore Mis-Attachment and Chromosome Mis-Segregation in Cancer Cells

PubMed Central

Silkworth, William T.; Nardi, Isaac K.; Scholl, Lindsey M.; Cimini, Daniela

2009-01-01

Many cancer cells display a CIN (Chromosome Instability) phenotype, by which they exhibit high rates of chromosome loss or gain at each cell cycle. Over the years, a number of different mechanisms, including mitotic spindle multipolarity, cytokinesis failure, and merotelic kinetochore orientation, have been proposed as causes of CIN. However, a comprehensive theory of how CIN is perpetuated is still lacking. We used CIN colorectal cancer cells as a model system to investigate the possible cellular mechanism(s) underlying CIN. We found that CIN cells frequently assembled multipolar spindles in early mitosis. However, multipolar anaphase cells were very rare, and live-cell experiments showed that almost all CIN cells divided in a bipolar fashion. Moreover, fixed-cell analysis showed high frequencies of merotelically attached lagging chromosomes in bipolar anaphase CIN cells, and higher frequencies of merotelic attachments in multipolar vs. bipolar prometaphases. Finally, we found that multipolar CIN prometaphases typically possessed γ-tubulin at all spindle poles, and that a significant fraction of bipolar metaphase/early anaphase CIN cells possessed more than one centrosome at a single spindle pole. Taken together, our data suggest a model by which merotelic kinetochore attachments can easily be established in multipolar prometaphases. Most of these multipolar prometaphase cells would then bi-polarize before anaphase onset, and the residual merotelic attachments would produce chromosome mis-segregation due to anaphase lagging chromosomes. We propose this spindle pole coalescence mechanism as a major contributor to chromosome instability in cancer cells. PMID:19668340

Analysis and Modeling of Chromosome Congression During Mitosis in the Chemotherapy Drug Cisplatin.

PubMed

Chacón, Jeremy M; Gardner, Melissa K

2013-12-01

The chemotherapy drug Cisplatin (cis-diamminedichloroplatinum(II)) induces crosslinks within and between DNA strands, and between DNA and nearby proteins. Therefore, Cisplatin-treated cells which progress into cell division may do so with altered chromosome mechanical properties. This could have important consequences for the successful completion of mitosis. Using Total Internal Reflection Fluorescence (TIRF) microscopy of live Cisplatin-treated Saccharomyces cerevisiae cells, we found that metaphase mitotic spindles have disorganized kinetochores relative to untreated cells, and also that there is increased variability in the chromosome stretching distance between sister centromeres. This suggests that chromosome stiffness may become more variable after Cisplatin treatment. We explored the effect of variable chromosome stiffness during mitosis using a stochastic model in which kinetochore microtubule dynamics were regulated by tension imparted by stretched sister chromosomes. Consistent with experimental results, increased variability of chromosome stiffness in the model led to disorganization of kinetochores in simulated metaphase mitotic spindles. Furthermore, the variability in simulated chromosome stretching tension was increased as chromosome stiffness became more variable. Because proper chromosome stretching tension may serve as a signal that is required for proper progression through mitosis, tension variability could act to impair this signal and thus prevent proper mitotic progression. Our results suggest a possible mitotic mode of action for the anti-cancer drug Cisplatin.

Chromosome congression by kinesin-5 motor-mediated disassembly of longer kinetochore microtubules

PubMed Central

Gardner, Melissa K; Bouck, David C.; Paliulis, Leocadia V.; Meehl, Janet B.; O’Toole, Eileen T.; Haase, Julian; Soubry, Adelheid; Joglekar, Ajit P.; Winey, Mark; Salmon, Edward D.; Bloom, Kerry; Odde, David J.

2008-01-01

Summary During mitosis, sister chromatids congress to the spindle equator and are subsequently segregated via attachment to dynamic kinetochore microtubule (kMT) plus-ends. A major question is how kMT plus-end assembly is spatially regulated to achieve chromosome congression. Here we find in budding yeast that the widely-conserved kinesin-5 sliding motor proteins, Cin8p and Kip1p, mediate chromosome congression by suppressing kMT plus-end assembly of longer kMTs. Of the two, Cin8p is the major effector and its activity requires a functional motor domain. In contrast, the depolymerizing kinesin-8 motor Kip3p plays a minor role in spatial regulation of yeast kMT assembly. Our analysis identified a model where kinesin-5 motors bind to kMTs, move to kMT plus ends, and upon arrival at a growing plus-end promote net kMT plus-end disassembly. In conclusion, we find that length-dependent control of net kMT assembly by kinesin-5 motors yields a simple and stable self-organizing mechanism for chromosome congression. PMID:19041752

Heterogeneous nuclear ribonucleoprotein K upregulates the kinetochore complex component NUF2 and promotes the tumorigenicity of colon cancer cells

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sugimasa, Hironobu; Taniue, Kenzui; Kurimoto, Akiko

2015-03-27

Heterogeneous nuclear ribonucleoprotein K (hnRNP K) is a multi-functional protein involved in transcription, mRNA splicing, mRNA stabilization and translation. Although hnRNP K has been suggested to play a role in the development of many cancers, its molecular function in colorectal cancer has remained elusive. Here we show that hnRNP K plays an important role in the mitotic process in HCT116 colon cancer cells. Furthermore, we demonstrate that hnRNP K directly transactivates the NUF2 gene, the product of which is a component of the NDC80 kinetochore complex and which is known to be critical for a stable spindle microtubule-kinetochore attachment. Inmore » addition, knockdown of both hnRNP K and NUF2 caused failure in metaphase chromosome alignment and drastic decrease in the growth of colon cancer cells. These results suggest that the hnRNP K-NUF2 axis is important for the mitotic process and proliferation of colon cancer cells and that this axis could be a target for the therapy of colon cancer. - Highlights: • hnRNP K is required for the tumorigenicity of colon cancer cells. • hnRNP K binds to the promoter region of NUF2 and activates its transcription. • NUF2 expression is correlated with hnRNP K expression in colorectal cancer tissue. • hnRNP K and NUF2 are required for metaphase chromosome alignment. • The hnRNP K-NUF2 axis is important for the proliferation of colon cancer cells.« less

Transcription and ncRNAs: at the cent(rome)re of kinetochore assembly and maintenance.

PubMed

Scott, Kristin C

2013-12-01

Centromeres are sites of chromosomal spindle attachment during mitosis and meiosis. Centromeres are defined, in part, by a distinct chromatin landscape in which histone H3 is replaced by the conserved histone H3 variant, CENP-A. Sequences competent for centromere formation and function vary among organisms and are typically composed of repetitive DNA. It is unclear how such diverse genomic signals are integrated with the epigenetic mechanisms that govern CENP-A incorporation at a single locus on each chromosome. Recent work highlights the intriguing possibility that the transcriptional properties of centromeric core DNA contribute to centromere identity and maintenance through cell division. Moreover, core-derived noncoding RNAs (ncRNAs) have emerged as active participants in the regulation and control of centromere activity in plants and mammals. This paper reviews the transcriptional properties of eukaryotic centromeres and discusses the known roles of core-derived ncRNAs in chromatin integrity, kinetochore assembly, and centromere activity.

NuSAP governs chromosome oscillation by facilitating the Kid-generated polar ejection force.

PubMed

Li, Chenyu; Xue, Chenyi; Yang, Qiaoyun; Low, Boon Chuan; Liou, Yih-Cherng

2016-02-03

In vertebrate cells, chromosomes oscillate to align precisely during metaphase. NuSAP, a microtubule-associated protein, plays a critical role in stabilizing spindle microtubules. In this study, we utilize 3D time-lapse live-cell imaging to monitor the role of NuSAP in chromosome oscillation and identify NuSAP as a novel regulator of the chromokinesin, Kid. Depletion of NuSAP significantly suppresses the amplitude and velocity of chromosome oscillation. We analyse the effects of NuSAP and Kid depletion in monopolar and bipolar cells with or without kinetochore microtubule depletion. Twelve postulated conditions are deciphered to reveal the contribution of NuSAP to the polar force generated at kinetochore microtubules and to the regulation of the polar ejection force generated by Kid, thus revealing a pivotal role of NuSAP in chromosome oscillation.

Sustained Mps1 activity is required in mitosis to recruit O-Mad2 to the Mad1-C-Mad2 core complex.

PubMed

Hewitt, Laura; Tighe, Anthony; Santaguida, Stefano; White, Anne M; Jones, Clifford D; Musacchio, Andrea; Green, Stephen; Taylor, Stephen S

2010-07-12

Mps1 is an essential component of the spindle assembly checkpoint. In this study, we describe a novel Mps1 inhibitor, AZ3146, and use it to probe the role of Mps1's catalytic activity during mitosis. When Mps1 is inhibited before mitotic entry, subsequent recruitment of Mad1 and Mad2 to kinetochores is abolished. However, if Mps1 is inhibited after mitotic entry, the Mad1-C-Mad2 core complex remains kinetochore bound, but O-Mad2 is not recruited to the core. Although inhibiting Mps1 also interferes with chromosome alignment, we see no obvious effect on aurora B activity. In contrast, kinetochore recruitment of centromere protein E (CENP-E), a kinesin-related motor protein, is severely impaired. Strikingly, inhibition of Mps1 significantly increases its own abundance at kinetochores. Furthermore, we show that Mps1 can dimerize and transphosphorylate in cells. We propose a model whereby Mps1 transphosphorylation results in its release from kinetochores, thus facilitating recruitment of O-Mad2 and CENP-E and thereby simultaneously promoting checkpoint signaling and chromosome congression.

Dynamic autophosphorylation of mps1 kinase is required for faithful mitotic progression.

PubMed

Wang, Xinghui; Yu, Huijuan; Xu, Leilei; Zhu, Tongge; Zheng, Fan; Fu, Chuanhai; Wang, Zhiyong; Dou, Zhen

2014-01-01

The spindle assembly checkpoint (SAC) is a surveillance mechanism monitoring cell cycle progression, thus ensuring accurate chromosome segregation. The conserved mitotic kinase Mps1 is a key component of the SAC. The human Mps1 exhibits comprehensive phosphorylation during mitosis. However, the related biological relevance is largely unknown. Here, we demonstrate that 8 autophosphorylation sites within the N-terminus of Mps1, outside of the catalytic domain, are involved in regulating Mps1 kinetochore localization. The phospho-mimicking mutant of the 8 autophosphorylation sites impairs Mps1 localization to kinetochore and also affects the kinetochore recruitment of BubR1 and Mad2, two key SAC effectors, subsequently leading to chromosome segregation errors. Interestingly, the non-phosphorylatable mutant of the 8 autophosphorylation sites enhances Mps1 kinetochore localization and delays anaphase onset. We further show that the Mps1 phospho-mimicking and non-phosphorylatable mutants do not affect metaphase chromosome congression. Thus, our results highlight the importance of dynamic autophosphorylation of Mps1 in regulating accurate chromosome segregation and ensuring proper mitotic progression.

Dynamic Autophosphorylation of Mps1 Kinase Is Required for Faithful Mitotic Progression

PubMed Central

Wang, Xinghui; Yu, Huijuan; Xu, Leilei; Zhu, Tongge; Zheng, Fan; Fu, Chuanhai; Wang, Zhiyong; Dou, Zhen

2014-01-01

The spindle assembly checkpoint (SAC) is a surveillance mechanism monitoring cell cycle progression, thus ensuring accurate chromosome segregation. The conserved mitotic kinase Mps1 is a key component of the SAC. The human Mps1 exhibits comprehensive phosphorylation during mitosis. However, the related biological relevance is largely unknown. Here, we demonstrate that 8 autophosphorylation sites within the N-terminus of Mps1, outside of the catalytic domain, are involved in regulating Mps1 kinetochore localization. The phospho-mimicking mutant of the 8 autophosphorylation sites impairs Mps1 localization to kinetochore and also affects the kinetochore recruitment of BubR1 and Mad2, two key SAC effectors, subsequently leading to chromosome segregation errors. Interestingly, the non-phosphorylatable mutant of the 8 autophosphorylation sites enhances Mps1 kinetochore localization and delays anaphase onset. We further show that the Mps1 phospho-mimicking and non-phosphorylatable mutants do not affect metaphase chromosome congression. Thus, our results highlight the importance of dynamic autophosphorylation of Mps1 in regulating accurate chromosome segregation and ensuring proper mitotic progression. PMID:25265012

NuSAP governs chromosome oscillation by facilitating the Kid-generated polar ejection force

PubMed Central

Li, Chenyu; Xue, Chenyi; Yang, Qiaoyun; Low, Boon Chuan; Liou, Yih-Cherng

2016-01-01

In vertebrate cells, chromosomes oscillate to align precisely during metaphase. NuSAP, a microtubule-associated protein, plays a critical role in stabilizing spindle microtubules. In this study, we utilize 3D time-lapse live-cell imaging to monitor the role of NuSAP in chromosome oscillation and identify NuSAP as a novel regulator of the chromokinesin, Kid. Depletion of NuSAP significantly suppresses the amplitude and velocity of chromosome oscillation. We analyse the effects of NuSAP and Kid depletion in monopolar and bipolar cells with or without kinetochore microtubule depletion. Twelve postulated conditions are deciphered to reveal the contribution of NuSAP to the polar force generated at kinetochore microtubules and to the regulation of the polar ejection force generated by Kid, thus revealing a pivotal role of NuSAP in chromosome oscillation. PMID:26839278

An overview of plant centromeres.

PubMed

Wang, Guixiang; Zhang, Xueyong; Jin, Weiwei

2009-09-01

The centromere is a defining region that mediates chromosome attachment to kinetochore microtubules and proper segregation of the sister chromatids. Intriguingly, satellite DNA and centromeric retrotransposon as major DNA constituents of centromere showed baffling diversification and species-specific. However, the key kinetochore proteins are conserved in both plants and animals, particularly the centromere-specific histone H3-like protein (CENH3) in all functional centromeres. Recent studies have highlighted the importance of epigenetic mechanisms in the establishment and maintenance of centromere identity. Here, we review the progress and compendium of research on plant centromere in the light of recent data.

Arabidopsis MZT1 homologs GIP1 and GIP2 are essential for centromere architecture.

PubMed

Batzenschlager, Morgane; Lermontova, Inna; Schubert, Veit; Fuchs, Jörg; Berr, Alexandre; Koini, Maria A; Houlné, Guy; Herzog, Etienne; Rutten, Twan; Alioua, Abdelmalek; Fransz, Paul; Schmit, Anne-Catherine; Chabouté, Marie-Edith

2015-07-14

Centromeres play a pivotal role in maintaining genome integrity by facilitating the recruitment of kinetochore and sister-chromatid cohesion proteins, both required for correct chromosome segregation. Centromeres are epigenetically specified by the presence of the histone H3 variant (CENH3). In this study, we investigate the role of the highly conserved γ-tubulin complex protein 3-interacting proteins (GIPs) in Arabidopsis centromere regulation. We show that GIPs form a complex with CENH3 in cycling cells. GIP depletion in the gip1gip2 knockdown mutant leads to a decreased CENH3 level at centromeres, despite a higher level of Mis18BP1/KNL2 present at both centromeric and ectopic sites. We thus postulate that GIPs are required to ensure CENH3 deposition and/or maintenance at centromeres. In addition, the recruitment at the centromere of other proteins such as the CENP-C kinetochore component and the cohesin subunit SMC3 is impaired in gip1gip2. These defects in centromere architecture result in aneuploidy due to severely altered centromeric cohesion. Altogether, we ascribe a central function to GIPs for the proper recruitment and/or stabilization of centromeric proteins essential in the specification of the centromere identity, as well as for centromeric cohesion in somatic cells.

Tension-Induced Error Correction and Not Kinetochore Attachment Status Activates the SAC in an Aurora-B/C-Dependent Manner in Oocytes.

PubMed

Vallot, Antoine; Leontiou, Ioanna; Cladière, Damien; El Yakoubi, Warif; Bolte, Susanne; Buffin, Eulalie; Wassmann, Katja

2018-01-08

Cell division with partitioning of the genetic material should take place only when paired chromosomes named bivalents (meiosis I) or sister chromatids (mitosis and meiosis II) are correctly attached to the bipolar spindle in a tension-generating manner. For this to happen, the spindle assembly checkpoint (SAC) checks whether unattached kinetochores are present, in which case anaphase onset is delayed to permit further establishment of attachments. Additionally, microtubules are stabilized when they are attached and under tension. In mitosis, attachments not under tension activate the so-named error correction pathway depending on Aurora B kinase substrate phosphorylation. This leads to microtubule detachments, which in turn activates the SAC [1-3]. Meiotic divisions in mammalian oocytes are highly error prone, with severe consequences for fertility and health of the offspring [4, 5]. Correct attachment of chromosomes in meiosis I leads to the generation of stretched bivalents, but-unlike mitosis-not to tension between sister kinetochores, which co-orient. Here, we set out to address whether reduction of tension applied by the spindle on bioriented bivalents activates error correction and, as a consequence, the SAC. Treatment of oocytes in late prometaphase I with Eg5 kinesin inhibitor affects spindle tension, but not attachments, as we show here using an optimized protocol for confocal imaging. After Eg5 inhibition, bivalents are correctly aligned but less stretched, and as a result, Aurora-B/C-dependent error correction with microtubule detachment takes place. This loss of attachments leads to SAC activation. Crucially, SAC activation itself does not require Aurora B/C kinase activity in oocytes. Copyright © 2017 Elsevier Ltd. All rights reserved.

STAG2 promotes error correction in mitosis by regulating kinetochore-microtubule attachments.

PubMed

Kleyman, Marianna; Kabeche, Lilian; Compton, Duane A

2014-10-01

Mutations in the STAG2 gene are present in ∼20% of tumors from different tissues of origin. STAG2 encodes a subunit of the cohesin complex, and tumors with loss-of-function mutations are usually aneuploid and display elevated frequencies of lagging chromosomes during anaphase. Lagging chromosomes are a hallmark of chromosomal instability (CIN) arising from persistent errors in kinetochore-microtubule (kMT) attachment. To determine whether the loss of STAG2 increases the rate of formation of kMT attachment errors or decreases the rate of their correction, we examined mitosis in STAG2-deficient cells. STAG2 depletion does not impair bipolar spindle formation or delay mitotic progression. Instead, loss of STAG2 permits excessive centromere stretch along with hyperstabilization of kMT attachments. STAG2-deficient cells display mislocalization of Bub1 kinase, Bub3 and the chromosome passenger complex. Importantly, strategically destabilizing kMT attachments in tumor cells harboring STAG2 mutations by overexpression of the microtubule-destabilizing enzymes MCAK (also known as KIF2C) and Kif2B decreased the rate of lagging chromosomes and reduced the rate of chromosome missegregation. These data demonstrate that STAG2 promotes the correction of kMT attachment errors to ensure faithful chromosome segregation during mitosis. © 2014. Published by The Company of Biologists Ltd.

The Argonaute CSR-1 and its 22G-RNA cofactors are required for holocentric chromosome segregation.

PubMed

Claycomb, Julie M; Batista, Pedro J; Pang, Ka Ming; Gu, Weifeng; Vasale, Jessica J; van Wolfswinkel, Josien C; Chaves, Daniel A; Shirayama, Masaki; Mitani, Shohei; Ketting, René F; Conte, Darryl; Mello, Craig C

2009-10-02

RNAi-related pathways regulate diverse processes, from developmental timing to transposon silencing. Here, we show that in C. elegans the Argonaute CSR-1, the RNA-dependent RNA polymerase EGO-1, the Dicer-related helicase DRH-3, and the Tudor-domain protein EKL-1 localize to chromosomes and are required for proper chromosome segregation. In the absence of these factors chromosomes fail to align at the metaphase plate and kinetochores do not orient to opposing spindle poles. Surprisingly, the CSR-1-interacting small RNAs (22G-RNAs) are antisense to thousands of germline-expressed protein-coding genes. Nematodes assemble holocentric chromosomes in which continuous kinetochores must span the expressed domains of the genome. We show that CSR-1 interacts with chromatin at target loci but does not downregulate target mRNA or protein levels. Instead, our findings support a model in which CSR-1 complexes target protein-coding domains to promote their proper organization within the holocentric chromosomes of C. elegans.

Spatial Rule-Based Modeling: A Method and Its Application to the Human Mitotic Kinetochore

PubMed Central

Ibrahim, Bashar; Henze, Richard; Gruenert, Gerd; Egbert, Matthew; Huwald, Jan; Dittrich, Peter

2013-01-01

A common problem in the analysis of biological systems is the combinatorial explosion that emerges from the complexity of multi-protein assemblies. Conventional formalisms, like differential equations, Boolean networks and Bayesian networks, are unsuitable for dealing with the combinatorial explosion, because they are designed for a restricted state space with fixed dimensionality. To overcome this problem, the rule-based modeling language, BioNetGen, and the spatial extension, SRSim, have been developed. Here, we describe how to apply rule-based modeling to integrate experimental data from different sources into a single spatial simulation model and how to analyze the output of that model. The starting point for this approach can be a combination of molecular interaction data, reaction network data, proximities, binding and diffusion kinetics and molecular geometries at different levels of detail. We describe the technique and then use it to construct a model of the human mitotic inner and outer kinetochore, including the spindle assembly checkpoint signaling pathway. This allows us to demonstrate the utility of the procedure, show how a novel perspective for understanding such complex systems becomes accessible and elaborate on challenges that arise in the formulation, simulation and analysis of spatial rule-based models. PMID:24709796

Feeling-of-knowing for proper names.

PubMed

Izaute, Marie; Chambres, Patrick; Larochelle, Serge

2002-12-01

The main objective of the presented study was to study feeling-of-knowing (FOK) in proper name retrieval. Many studies show that FOK can predict performance on a subsequent criterion test. Although feeling-of-knowing studies involve questions about proper names, none make this distinction between proper names and common names. Nevertheless, the specific character of proper names as a unique label referring to a person should allow participants to target precisely the desired verbal label. Our idea here was that the unique character of proper name information should result in more accurate FOK evaluations. In the experiment, participants evaluated feeling-of-knowing for proper and common name descriptions. The study demonstrates that FOK judgments are more accurate for proper names than for common names. The implications of the findings for proper names are briefly discussed in terms of feeling-of-knowing hypotheses.

Chromatin signaling to kinetochores: Trans-regulation of Dam1 methylation by histone H2B ubiquitination

PubMed Central

Latham, John A.; Chosed, Renée J.; Wang, Shanzhi; Dent, Sharon Y.R.

2011-01-01

Summary Histone H3K4 trimethylation by the Set1/MLL family of proteins provides a hallmark for transcriptional activity from yeast to humans. In S. cerevisiae, H3K4 methylation is mediated by the Set1-containing COMPASS complex and is regulated in trans by prior ubiquitination of histone H2BK123. All of the events that regulate H2BK123ub and H3K4me are thought to occur at gene promoters. Here we report that this pathway is indispensable for methylation of the only other known substrate of Set1, K233 in Dam1, at kinetochores. Deletion of RAD6, BRE1, or Paf1 complex members abolishes Dam1 methylation, as does mutation of H2BK123. Our results demonstrate that Set1-mediated methylation is regulated by a general pathway regardless of substrate that is composed of transcriptional regulatory factors functioning independently of transcription. Moreover, our data identify a node of regulatory cross-talk in trans between a histone modification and modification on a non-histone protein, demonstrating that changing chromatin states can signal functional changes in other essential cellular proteins and machineries. PMID:21884933

Diversity in Requirement of Genetic and Epigenetic Factors for Centromere Function in Fungi ▿

PubMed Central

Roy, Babhrubahan; Sanyal, Kaustuv

2011-01-01

A centromere is a chromosomal region on which several proteins assemble to form the kinetochore. The centromere-kinetochore complex helps in the attachment of chromosomes to spindle microtubules to mediate segregation of chromosomes to daughter cells during mitosis and meiosis. In several budding yeast species, the centromere forms in a DNA sequence-dependent manner, whereas in most other fungi, factors other than the DNA sequence also determine the centromere location, as centromeres were able to form on nonnative sequences (neocentromeres) when native centromeres were deleted in engineered strains. Thus, in the absence of a common DNA sequence, the cues that have facilitated centromere formation on a specific DNA sequence for millions of years remain a mystery. Kinetochore formation is facilitated by binding of a centromere-specific histone protein member of the centromeric protein A (CENP-A) family that replaces a canonical histone H3 to form a specialized centromeric chromatin structure. However, the process of kinetochore formation on the rapidly evolving and seemingly diverse centromere DNAs in different fungal species is largely unknown. More interestingly, studies in various yeasts suggest that the factors required for de novo centromere formation (establishment) may be different from those required for maintenance (propagation) of an already established centromere. Apart from the DNA sequence and CENP-A, many other factors, such as posttranslational modification (PTM) of histones at centric and pericentric chromatin, RNA interference, and DNA methylation, are also involved in centromere formation, albeit in a species-specific manner. In this review, we discuss how several genetic and epigenetic factors influence the evolution of structure and function of centromeres in fungal species. PMID:21908596

Mold Allergy: Proper Humidifier Care

MedlinePlus

... Training Home Conditions Allergy Allergy Overview Allergy Allergens Mold Allergy Proper Humidifier Care Proper Humidifier Care Make ... neglected humidifier can be a major source of mold and mold spores. Learn how to keep a ...

PinX1 is recruited to the mitotic chromosome periphery by Nucleolin and facilitates chromosome congression.

PubMed

Li, Na; Yuan, Kai; Yan, Feng; Huo, Yuda; Zhu, Tongge; Liu, Xing; Guo, Zhen; Yao, Xuebiao

2009-06-19

Mitotic chromosome movements are orchestrated by interactions between spindle microtubules and chromosomes. It is well known that kinetochore is the major site where microtubule-chromosome attachment occurs. However, the functions of other domains of chromosome such as chromosome periphery have remained elusive. Our previous studies show that PinX1 distributes to chromosome periphery and kinetochore during mitosis, and harbors the microtubule binding activity. Here we report that PinX1 interacts with Nucleolin, a chromosome periphery protein, through its C-termini. Deconvolution microscopic analyses show PinX1 mainly co-localizes with Nucleolin at chromosome periphery in prometaphase. Moreover, depletion of Nucleolin abolishes chromosome periphery localizations of PinX1, suggesting a functional interrelationship between PinX1 and Nucleolin. Importantly, repression of PinX1 and Nucleolin abrogates chromosome segregation in real-time mitosis, validating the functional importance of PinX1-Nucleolin interaction. We propose PinX1 is recruited to chromosome periphery by Nucleolin and a complex of PinX1 and Nucleolin is essential for faithful chromosome congression.

7 CFR 29.112 - Proper light.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 7 Agriculture 2 2010-01-01 2010-01-01 false Proper light. 29.112 Section 29.112 Agriculture... INSPECTION Regulations Inspectors, Samplers, and Weighers § 29.112 Proper light. Tobacco shall not be inspected or sampled for the purposes of the Act except when displayed in proper light for correct...

7 CFR 29.112 - Proper light.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 7 Agriculture 2 2014-01-01 2014-01-01 false Proper light. 29.112 Section 29.112 Agriculture... INSPECTION Regulations Inspectors, Samplers, and Weighers § 29.112 Proper light. Tobacco shall not be inspected or sampled for the purposes of the Act except when displayed in proper light for correct...

7 CFR 29.112 - Proper light.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 7 Agriculture 2 2013-01-01 2013-01-01 false Proper light. 29.112 Section 29.112 Agriculture... INSPECTION Regulations Inspectors, Samplers, and Weighers § 29.112 Proper light. Tobacco shall not be inspected or sampled for the purposes of the Act except when displayed in proper light for correct...

7 CFR 29.112 - Proper light.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 7 Agriculture 2 2011-01-01 2011-01-01 false Proper light. 29.112 Section 29.112 Agriculture... INSPECTION Regulations Inspectors, Samplers, and Weighers § 29.112 Proper light. Tobacco shall not be inspected or sampled for the purposes of the Act except when displayed in proper light for correct...

7 CFR 29.112 - Proper light.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 7 Agriculture 2 2012-01-01 2012-01-01 false Proper light. 29.112 Section 29.112 Agriculture... INSPECTION Regulations Inspectors, Samplers, and Weighers § 29.112 Proper light. Tobacco shall not be inspected or sampled for the purposes of the Act except when displayed in proper light for correct...

Proper Acknowledgment?

ERIC Educational Resources Information Center

East, Julianne

2005-01-01

The concern in Australian universities about the prevalence of plagiarism has led to the development of policies about academic integrity and in turn focused attention on the need to inform students about how to avoid plagiarism and how to properly acknowledge. Teaching students how to avoid plagiarism can appear to be straightforward if based on…

Promoting Proper Education for Sustainability: An Exploratory Study of ICT Enhanced Problem Based Learning in a Developing Country

ERIC Educational Resources Information Center

Roy, Amit; Kihoza, Patrick; Suhonen, Jarkko; Vesisenaho, Mikko; Tukiaianen, Markku

2014-01-01

One of the goals of education is to create responsible citizens who can adequately understand the problems faced by their societies and who can then act to help solve them. Such behaviour can be fostered through proper education that facilitates expert knowledge about social issues, nurtures the ability to think critically and grows the skills…

Proper Names: Reference and Attribution

ERIC Educational Resources Information Center

Maumus, Michael Fletcher

2012-01-01

In the wake of Saul Kripke's landmark "Naming and Necessity," the claim that proper names are directly referential expressions devoid of descriptive content has come to verge on philosophical commonplace. Nevertheless, the return to a purely referential semantics for proper names has coincided with the resurgence of the very puzzles…

The insecticide buprofezin induces morphological transformation and kinetochore-positive micronuclei in cultured Syrian hamster embryo cells in the absence of detectable DNA damage.

PubMed

Herrera, L A; Ostrosky-Wegman, P; Schiffmann, D; Chen, Q Y; Ziegler-Skylakakis, K; Andrae, U

1993-11-01

The insecticide buprofezin was examined for its genotoxicity in cultured Syrian hamster embryo cells in order to better understand the mechanisms underlying the genotoxicity of the compound in mammalian cells. Exposure to buprofezin concentrations of 12.5-100 microM did not significantly affect the colony-forming ability of the cells, but did result in increased frequencies of morphologically transformed colonies. Treatment with buprofezin did not cause a detectable induction of DNA repair synthesis, an indicator of DNA damage, but significantly increased the frequency of micronuclei. Immunostaining of the cells with antikinetochore antibody (CREST antibody) showed that essentially all of the buprofezin-induced micronuclei were kinetochore-positive. The results suggest that morphological transformation of Syrian hamster embryo cells by buprofezin results from an interaction of the compound or a metabolite of it with the mitotic apparatus rather than from DNA damage.

7 CFR 993.21a - Proper storage.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 7 Agriculture 8 2013-01-01 2013-01-01 false Proper storage. 993.21a Section 993.21a Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (MARKETING AGREEMENTS... Order Regulating Handling Definitions § 993.21a Proper storage. Proper storage means storage of such...

7 CFR 993.21a - Proper storage.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 7 Agriculture 8 2011-01-01 2011-01-01 false Proper storage. 993.21a Section 993.21a Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (Marketing Agreements... Order Regulating Handling Definitions § 993.21a Proper storage. Proper storage means storage of such...

7 CFR 993.21a - Proper storage.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 7 Agriculture 8 2012-01-01 2012-01-01 false Proper storage. 993.21a Section 993.21a Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (Marketing Agreements... Order Regulating Handling Definitions § 993.21a Proper storage. Proper storage means storage of such...

7 CFR 993.21a - Proper storage.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 7 Agriculture 8 2014-01-01 2014-01-01 false Proper storage. 993.21a Section 993.21a Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (MARKETING AGREEMENTS... Order Regulating Handling Definitions § 993.21a Proper storage. Proper storage means storage of such...

7 CFR 993.21a - Proper storage.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 7 Agriculture 8 2010-01-01 2010-01-01 false Proper storage. 993.21a Section 993.21a Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (Marketing Agreements... Order Regulating Handling Definitions § 993.21a Proper storage. Proper storage means storage of such...

Push or Pull? -- Cryo-Electron Microscopy of Microtubule's Dynamic Instability and Its Roles in the Kinetochore

NASA Astrophysics Data System (ADS)

Wang, Hong-Wei

2009-03-01

Microtubule is a biopolymer made up of alpha-beta-tubulin heterodimers. The tubulin dimers assemble head-to-tail as protofilaments and about 13 protofilaments interact laterally to form a hollow cylindrical structure which is the microtubule. As the major cytoskeleton in all eukaryotic cells, microtubules have the intrinsic property to switch stochastically between growth and shrinkage phases, a phenomenon termed as their dynamic instability. Microtubule's dynamic instability is closely related to the types of nucleotide (GTP or GDP) that binds to the beta-tubulin. We have biochemically trapped two types of assembly states of tubulin with GTP or GDP bound representing the polymerizing and depolymerizing ends of microtubules respectively. Using cryo-electron microscopy, we have elucidated the structures of these intermediate assemblies, showing that tubulin protofilaments demonstrate various curvatures and form different types of lateral interactions depending on the nucleotide states of tubulin and the temperature. Our work indicates that during the microtubule's dynamic cycle, tubulin undergoes various assembly states. These states, different from the straight microtubule, lend the highly dynamic and complicated behavior of microtubules. Our study of microtubule's interaction with certain kinetochore complexes suggests that the intermediate assemblies are responsible for specific mechanical forces that are required during the mitosis or meiosis. Our discoveries strongly suggest that a microtubule is a molecular machine rather than a simple cellular scaffold.

Deposition, turnover, and release of CENH3 at Arabidopsis centromeres.

PubMed

Lermontova, Inna; Rutten, Twan; Schubert, Ingo

2011-12-01

The kinetochore is a complex multiprotein structure located at centromeres and required for the proper segregation of chromosomes during mitosis and meiosis. An important role in kinetochore assembly and function plays the centromeric histone H3 variant (CENH3). Cell cycle stage of CENH3 deposition to centromeres varies between different organisms. We confirmed by in vivo studies that deposition of Arabidopsis CENH3 takes place at centromeres during G2 and demonstrated that additionally a low turnover of CENH3 occurs along the cell cycle, apparently for replacement of damaged protein. Furthermore, enhanced yellow fluorescent protein (EYFP)-CENH3 of photobleached chromocenters is not replaced by EYFP-CENH3 molecules from unbleached centromeres of the same nucleus, indicating a stable incorporation of CENH3 into centromeric nucleosomes. In differentiated endopolyploid nuclei however, the amount of CENH3 at centromeres declines with age.

Small-molecule kinase inhibitors provide insight into Mps1 cell cycle function.

PubMed

Kwiatkowski, Nicholas; Jelluma, Nannette; Filippakopoulos, Panagis; Soundararajan, Meera; Manak, Michael S; Kwon, Mijung; Choi, Hwan Geun; Sim, Taebo; Deveraux, Quinn L; Rottmann, Sabine; Pellman, David; Shah, Jagesh V; Kops, Geert J P L; Knapp, Stefan; Gray, Nathanael S

2010-05-01

Mps1, a dual-specificity kinase, is required for the proper functioning of the spindle assembly checkpoint and for the maintenance of chromosomal stability. As Mps1 function has been implicated in numerous phases of the cell cycle, the development of a potent, selective small-molecule inhibitor of Mps1 should facilitate dissection of Mps1-related biology. We describe the cellular effects and Mps1 cocrystal structures of new, selective small-molecule inhibitors of Mps1. Consistent with RNAi studies, chemical inhibition of Mps1 leads to defects in Mad1 and Mad2 establishment at unattached kinetochores, decreased Aurora B kinase activity, premature mitotic exit and gross aneuploidy, without any evidence of centrosome duplication defects. However, in U2OS cells having extra centrosomes (an abnormality found in some cancers), Mps1 inhibition increases the frequency of multipolar mitoses. Lastly, Mps1 inhibitor treatment resulted in a decrease in cancer cell viability.

Proper Accounting for Surface Area to Solution Volume Ratios in Exaggerated Extractions.

PubMed

Jenke, Dennis R; Rabinow, Barrett E

2017-01-01

When drug products contact plastic manufacturing components, packaging systems, and/or delivery devices, leachables from the plastics can accumulate in the drug product, potentially affecting its key quality attributes. Given practical issues associated with screening drug products for leachables, potential leachables are frequently surfaced as extractables revealed in extraction studies. To facilitate extractables discovery and identification and to shorten extraction times, extraction studies can be exaggerated and/or accelerated. One means of exaggerating an extraction is to increase the test article's extracted surface area to extraction solution volume ratio (SA/V), as it is generally accepted that an extractable's concentration in an extract is proportional to SA/V in a 1 to 1 manner. However, as the relationship between an extractable's concentration and SA/V depends on the extractable's plastic/solvent partition coefficient (k p/l ), the effect of SA/V on the extractable's concentrations can be either under- or over-estimated if a 1 to 1 proportion is used. This article presents the theoretical relationship between SA/V, concentration, and k p/l ; illustrates theory with a case study; and suggests proper exaggeration strategies. LAY ABSTRACT: When drug products are manufactured, stored, or delivered in systems that contain plastics, substances can be leached from the plastics and remain in the drug product, where they might affect the product's key quality attributes. To discover and identify these leached substances, the plastics are extracted under laboratory conditions and the extracts are appropriately tested. To facilitate this process, extracts may be generated under laboratory conditions that exaggerate or accelerate the drug product's clinical conditions of manufacturing or use. The proper use of the ratio of the extracted item's surface area to the volume of the extracting solution as an exaggeration parameter is discussed in this paper. © PDA

Proper Names a Cognitive-Philosophical Study

ERIC Educational Resources Information Center

Garcia-Ramirez, Eduardo

2010-01-01

Proper Names appear at the heart of several debates in philosophy and the cognitive sciences. These include "reference", "intentionality", and the nature of "belief" as well as "language acquisition", "cognitive development", and "memory". This dissertation follows a cognitive approach to the philosophical problems posed by proper names. It puts…

L1 Use in L2 Vocabulary Learning: Facilitator or Barrier

ERIC Educational Resources Information Center

Liu, Jing

2008-01-01

Based on empirical research and qualitative analysis, this paper aims to explore the effects of L1 use on L2 vocabulary teaching. The results show that, during L2 vocabulary teaching process, the proper application of L1 can effectively facilitate the memorization of new words, and the bilingual method (both English explanation and Chinese…

OH maser proper motions in Cepheus A

NASA Astrophysics Data System (ADS)

Migenes, V.; Cohen, R. J.; Brebner, G. C.

1992-02-01

MERLIN measurements made between 1982 and 1989 reveal proper motions of OH masers in the source Cepheus A. The proper motions are typically a few milliarcsec per year, and are mainly directed away from the central H II regions. Statistical analysis of the data suggests an expansion time-scale of some 300 yr. The distance of the source implied by the proper motions is 320+140/-80 pc, assuming that the expansion is isotropic. The proper motions can be reconciled with the larger distance of 730 pc which is generally accepted, provided that the masers are moving at large angles to the line of sight. The expansion time-scale agrees with that of the magnetic field decay recently reported by Cohen, et al. (1990).

Determination of proper motions in the Pleiades cluster

NASA Astrophysics Data System (ADS)

Schilbach, E.

1991-04-01

For 458 stars in the Pleiades field from the catalog of Eichhorn et al. (1970) proper motions were derived on Tautenburg and CERGA Schmidt telescope plates measured with the automated measuring machine MAMA in Paris. The catalog positions were considered as first epoch coordinates with an epoch difference of ca. 33 years to the observations. The results show good coincidence of proper motions derived with both Schmidt telescopes within the error bars. Comparison with proper motions determined by Vasilevskis et al. (1979) displays some significant differences but no systematic effects depending on plate coordinates or magnitudes could be found. An accuracy of 0.3 arcsec/100a for one proper motion component was estimated. According to the criterion of common proper motion 34 new cluster members were identified.

Using Technology to Facilitate Collaboration in Community-Based Participatory Research (CBPR)

PubMed Central

Jessell, Lauren; Smith, Vivian; Jemal, Alexis; Windsor, Liliane

2017-01-01

This study explores the use of Computer-Supported Collaborative Work (CSCW) technologies, by way of a computer-based system called iCohere. This system was used to facilitate collaboration conducting Community-Based Participatory Research (CBPR). Data was gathered from 13 members of a Community Collaborative Board (CCB). Analysis revealed that iCohere served the following functions: facilitating communication, providing a depository for information and resource sharing, and allowing for remote meeting attendance. Results indicated that while iCohere was useful in performing these functions, less expensive technologies had the potential to achieve similar goals if properly implemented. Implications for future research on CSCW systems and CBPR are discussed. PMID:29056871

Computation of Asteroid Proper Elements: Recent Advances

NASA Astrophysics Data System (ADS)

Knežević, Z.

2017-12-01

The recent advances in computation of asteroid proper elements are briefly reviewed. Although not representing real breakthroughs in computation and stability assessment of proper elements, these advances can still be considered as important improvements offering solutions to some practical problems encountered in the past. The problem of getting unrealistic values of perihelion frequency for very low eccentricity orbits is solved by computing frequencies using the frequency-modified Fourier transform. The synthetic resonant proper elements adjusted to a given secular resonance helped to prove the existence of Astraea asteroid family. The preliminary assessment of stability with time of proper elements computed by means of the analytical theory provides a good indication of their poorer performance with respect to their synthetic counterparts, and advocates in favor of ceasing their regular maintenance; the final decision should, however, be taken on the basis of more comprehensive and reliable direct estimate of their individual and sample average deviations from constancy.

Histone H2A is required for normal centromere function in Saccharomyces cerevisiae

PubMed Central

Pinto, Inés; Winston, Fred

2000-01-01

Histones are structural and functional components of the eukaryotic chromosome, and their function is essential for normal cell cycle progression. In this work, we describe the characterization of two Saccharomyces cerevisiae cold-sensitive histone H2A mutants. Both mutants contain single amino acid replacements of residues predicted to be on the surface of the nucleosome and in close contact with DNA. We show that these H2A mutations cause an increase-in-ploidy phenotype, an increased rate of chromosome loss, and a defect in traversing the G2–M phase of the cell cycle. Moreover, these H2A mutations show genetic interactions with mutations in genes encoding kinetochore components. Finally, chromatin analysis of these H2A mutants has revealed an altered centromeric chromatin structure. Taken together, these results strongly suggest that histone H2A is required for proper centromere–kinetochore function during chromosome segregation. PMID:10747028

PROPER: global protein interaction network alignment through percolation matching.

PubMed

Kazemi, Ehsan; Hassani, Hamed; Grossglauser, Matthias; Pezeshgi Modarres, Hassan

2016-12-12

The alignment of protein-protein interaction (PPI) networks enables us to uncover the relationships between different species, which leads to a deeper understanding of biological systems. Network alignment can be used to transfer biological knowledge between species. Although different PPI-network alignment algorithms were introduced during the last decade, developing an accurate and scalable algorithm that can find alignments with high biological and structural similarities among PPI networks is still challenging. In this paper, we introduce a new global network alignment algorithm for PPI networks called PROPER. Compared to other global network alignment methods, our algorithm shows higher accuracy and speed over real PPI datasets and synthetic networks. We show that the PROPER algorithm can detect large portions of conserved biological pathways between species. Also, using a simple parsimonious evolutionary model, we explain why PROPER performs well based on several different comparison criteria. We highlight that PROPER has high potential in further applications such as detecting biological pathways, finding protein complexes and PPI prediction. The PROPER algorithm is available at http://proper.epfl.ch .

Spinfoam cosmology with the proper vertex amplitude

NASA Astrophysics Data System (ADS)

Vilensky, Ilya

2017-11-01

The proper vertex amplitude is derived from the Engle-Pereira-Rovelli-Livine vertex by restricting to a single gravitational sector in order to achieve the correct semi-classical behaviour. We apply the proper vertex to calculate a cosmological transition amplitude that can be viewed as the Hartle-Hawking wavefunction. To perform this calculation we deduce the integral form of the proper vertex and use extended stationary phase methods to estimate the large-volume limit. We show that the resulting amplitude satisfies an operator constraint whose classical analogue is the Hamiltonian constraint of the Friedmann-Robertson-Walker cosmology. We find that the constraint dynamically selects the relevant family of coherent states and demonstrate a similar dynamic selection in standard quantum mechanics. We investigate the effects of dynamical selection on long-range correlations.

CENP-E Kinesin Interacts with SKAP Protein to Orchestrate Accurate Chromosome Segregation in Mitosis*

PubMed Central

Huang, Yuejia; Wang, Wenwen; Yao, Phil; Wang, Xiwei; Liu, Xing; Zhuang, Xiaoxuan; Yan, Feng; Zhou, Jinhua; Du, Jian; Ward, Tarsha; Zou, Hanfa; Zhang, Jiancun; Fang, Guowei; Ding, Xia; Dou, Zhen; Yao, Xuebiao

2012-01-01

Mitotic chromosome segregation is orchestrated by the dynamic interaction of spindle microtubules with the kinetochore. Although previous studies show that the mitotic kinesin CENP-E forms a link between attachment of the spindle microtubule to the kinetochore and the mitotic checkpoint signaling cascade, the molecular mechanism underlying dynamic kinetochore-microtubule interactions in mammalian cells remains elusive. Here, we identify a novel interaction between CENP-E and SKAP that functions synergistically in governing dynamic kinetochore-microtubule interactions. SKAP binds to the C-terminal tail of CENP-E in vitro and is essential for an accurate kinetochore-microtubule attachment in vivo. Immunoelectron microscopic analysis indicates that SKAP is a constituent of the kinetochore corona fibers of mammalian centromeres. Depletion of SKAP or CENP-E by RNA interference results in a dramatic reduction of inter-kinetochore tension, which causes chromosome mis-segregation with a prolonged delay in achieving metaphase alignment. Importantly, SKAP binds to microtubules in vitro, and this interaction is synergized by CENP-E. Based on these findings, we propose that SKAP cooperates with CENP-E to orchestrate dynamic kinetochore-microtubule interaction for faithful chromosome segregation. PMID:22110139

Proper projective symmetry in LRS Bianchi type V spacetimes

NASA Astrophysics Data System (ADS)

Shabbir, Ghulam; Mahomed, K. S.; Mahomed, F. M.; Moitsheki, R. J.

2018-04-01

In this paper, we investigate proper projective vector fields of locally rotationally symmetric (LRS) Bianchi type V spacetimes using direct integration and algebraic techniques. Despite the non-degeneracy in the Riemann tensor eigenvalues, we classify proper Bianchi type V spacetimes and show that the above spacetimes do not admit proper projective vector fields. Here, in all the cases projective vector fields are Killing vector fields.

Self-Optimized Biological Channels in Facilitating the Transmembrane Movement of Charged Molecules

PubMed Central

Huyen, V. T. N.; Lap, Vu Cong; Nguyen, V. Lien

2016-01-01

We consider an anisotropically two-dimensional diffusion of a charged molecule (particle) through a large biological channel under an external voltage. The channel is modeled as a cylinder of three structure parameters: radius, length, and surface density of negative charges located at the channel interior-lining. These charges induce inside the channel a potential that plays a key role in controlling the particle current through the channel. It was shown that to facilitate the transmembrane particle movement the channel should be reasonably self-optimized so that its potential coincides with the resonant one, resulting in a large particle current across the channel. Observed facilitation appears to be an intrinsic property of biological channels, regardless of the external voltage or the particle concentration gradient. This facilitation is very selective in the sense that a channel of definite structure parameters can facilitate the transmembrane movement of only particles of proper valence at corresponding temperatures. Calculations also show that the modeled channel is nonohmic with the ion conductance which exhibits a resonance at the same channel potential as that identified in the current. PMID:27022394

INTERNAL PROPER MOTIONS IN THE ESKIMO NEBULA

DOE Office of Scientific and Technical Information (OSTI.GOV)

García-Díaz, Ma. T.; Gutiérrez, L.; Steffen, W.

We present measurements of internal proper motions at more than 500 positions of NGC 2392, the Eskimo Nebula, based on images acquired with WFPC2 on board the Hubble Space Telescope at two epochs separated by 7.695 yr. Comparisons of the two observations clearly show the expansion of the nebula. We measured the amplitude and direction of the motion of local structures in the nebula by determining their relative shift during that interval. In order to assess the potential uncertainties in the determination of proper motions in this object, in general, the measurements were performed using two different methods, used previously in themore » literature. We compare the results from the two methods, and to perform the scientific analysis of the results we choose one, the cross-correlation method, because it is more reliable. We go on to perform a ''criss-cross'' mapping analysis on the proper motion vectors, which helps in the interpretation of the velocity pattern. By combining our results of the proper motions with radial velocity measurements obtained from high resolution spectroscopic observations, and employing an existing 3D model, we estimate the distance to the nebula to be 1.3 kpc.« less

32 CFR 842.28 - Who are proper claimants.

Code of Federal Regulations, 2010 CFR

2010-07-01

... paid from appropriated funds. (c) DOD school teachers and school administrative personnel who are provided logistic and administrative support by an Air Force installation commander. (d) Air Force Reserve... ADMINISTRATIVE CLAIMS Personnel Claims (31 U.S.C. 3701, 3721) § 842.28 Who are proper claimants. Proper claimants...

Asteroid proper elements and secular resonances

NASA Technical Reports Server (NTRS)

Knezevic, Zoran; Milani, Andrea

1992-01-01

In a series of papers (e.g., Knezevic, 1991; Milani and Knezevic, 1990; 1991) we reported on the progress we were making in computing asteroid proper elements, both as regards their accuracy and long-term stability. Additionally, we reported on the efficiency and 'intelligence' of our software. At the same time, we studied the associated problems of resonance effects, and we introduced the new class of 'nonlinear' secular resonances; we determined the locations of these secular resonances in proper-element phase space and analyzed their impact on the asteroid family classification. Here we would like to summarize the current status of our work and possible further developments.

Proper laboratory notebook practices: protecting your intellectual property.

PubMed

Nickla, Jason T; Boehm, Matthew B

2011-03-01

A laboratory notebook contains a wealth of knowledge that can be critical for establishing evidence in support of intellectual property rights and for refuting claims of research misconduct. The proper type, organization, use, maintenance, and storage of laboratory notebooks should be a priority for everyone at research institutions. Failure to properly document research activities can lead to serious problems, including the loss of valuable patent rights. Consequences of improper laboratory notebook practices can be harsh; numerous examples are described in court cases and journal articles, indicating a need for research institutions to develop strict policies on the proper use and storage of research documentation.

Developing proper mealtime behaviors of the institutionalized retarded.

PubMed

O'brien, F; Azrin, N H

1972-01-01

The institutionalized mentally retarded display a variety of unsanitary, disruptive, and improper table manners. A program was developed that included (1) acquisition-training of a high standard of proper table manners and (2) maintenance procedures to provide continued motivation to maintain proper mealtime behaviors and decrease improper skills. Twelve retardates received acquisition training, individually, by a combination of verbal instruction, imitation, and manual guidance. The students then ate in their group dining arrangement where the staff supervisor provided continuing approval for proper manners and verbal correction and timeout for improper manners. The results were: (1) the trained retardates showed significant improvement, whereas those untrained did not; (2) the trained retardates ate as well in the institution as non-retarded customers did in a public restaurant; (3) proper eating was maintained in the group dining setting; (4) timeout was rarely needed; (5) the program was easily administered by regular staff in a regular dining setting. The rapidity, feasibility, and effectiveness of the program suggests the program as a solution to improper mealtime behaviors by the institutionalized mentally retarded.

Defining the Centromere.

ERIC Educational Resources Information Center

Erickson, John

1983-01-01

Focusing on the centromere (kinetochore), discusses what term should be used to represent this cellular component. Also discusses centromere/kinetochore replication, structure of the kinetochore, and the nature of the binding material that holds until anaphase of mitosis and meiosis. (JN)

Considerations for proper selection of dental cements.

PubMed

Simon, James F; Darnell, Laura A

2012-01-01

Selecting the proper cement for sufficient bond strength has become progressively complicated as the number of different materials for indirect restorations has increased. The success of any restoration is highly dependent on the proper cement being chosen and used. The function of the cement is not only to seal the restoration on the tooth but also, in some cases, to support the retention of the restoration. This ability to strengthen retention varies by the cement chosen by the clinician; therefore, careful consideration must precede cement selection.

Lick Northern Proper Motion Program: NPM2

NASA Astrophysics Data System (ADS)

Jones, B. F.; Hanson, R. B.; Klemola, A. R.

2000-05-01

The Lick Northern Proper Motion (NPM) program is nearing completion after a half-century of work. Two-epoch photography began in 1947 and was completed in 1988. Measurements and reductions for proper motions, positions, and two-color photometry in the sky outside the Milky Way (``NPM1'') began in 1975 and were completed in 1992. The Lick NPM1 Catalog, containing 149,000 stars, was distributed in 1993. Work on the Milky Way sky (``NPM2'') comprising some 300,000 stars, began in 1996, and plate measurements were finished in 1999. The NPM program will be completed with the publication of the Lick NPM2 Catalog in 2003. The NPM program will provide absolute proper motions, measured on an inertial system defined by some 50,000 faint galaxies, for over 400,000 stars from 9 < B < 18, covering the northern two-thirds of the sky. Included in the NPM catalogs are many stars of astrophysical interest, anonymous stars for galactic studies, and stars from positional catalogues and proper motion surveys. Current work at Lick encompasses data reductions and star identifications for NPM2. Procedures are based on NPM1, with appropriate modifications. Reference galaxies are not available in the Milky Way sky, so the Hipparcos Catalogue is used to link the NPM2 proper motions to the inertial system defined by NPM1. The large number of stars in NPM2 reflects the higher density of stars near the Galactic plane and toward the Galactic center. The NPM catalogs will have lasting value as a unique database for future studies in galactic structure, stellar kinematics, and astrometry. As we produce NPM2, we are also applying the NPM data to several outstanding problems in these research fields. We would like to thank Dave Monet and the USNO for measuring the NPM2 plates. We thank the National Science Foundation for its continued support of the NPM program. The work reported here was supported by NSF grant AST 9530632.

Proper-motion Study of the Magellanic Clouds Using SPM Material

NASA Astrophysics Data System (ADS)

Vieira, Katherine; Girard, Terrence M.; van Altena, William F.; Zacharias, Norbert; Casetti-Dinescu, Dana I.; Korchagin, Vladimir I.; Platais, Imants; Monet, David G.; López, Carlos E.; Herrera, David; Castillo, Danilo J.

2010-12-01

Absolute proper motions are determined for stars and galaxies to V = 17.5 over a 450 deg2 area that encloses both Magellanic Clouds. The proper motions are based on photographic and CCD observations of the Yale/San Juan Southern Proper Motion program, which span a baseline of 40 years. Multiple, local relative proper-motion measures are combined in an overlap solution using photometrically selected Galactic disk stars to define a global relative system that is then transformed to absolute using external galaxies and Hipparcos stars to tie into the ICRS. The resulting catalog of 1.4 million objects is used to derive the mean absolute proper motions of the Large Magellanic Cloud (LMC) and the Small Magellanic Cloud (SMC); (μαcos δ, μδ)LMC = (1.89, + 0.39) ± (0.27, 0.27) masyr-1 and (μαcos δ, μδ)SMC = (0.98, - 1.01) ± (0.30, 0.29) masyr-1. These mean motions are based on best-measured samples of 3822 LMC stars and 964 SMC stars. A dominant portion (0.25 mas yr-1) of the formal errors is due to the estimated uncertainty in the inertial system of the Hipparcos Catalog stars used to anchor the bright end of our proper motion measures. A more precise determination can be made for the proper motion of the SMC relative to the LMC; (μαcos δ, μδ)SMC-LMC = (-0.91, - 1.49) ± (0.16, 0.15) masyr-1. This differential value is combined with measurements of the proper motion of the LMC taken from the literature to produce new absolute proper-motion determinations for the SMC, as well as an estimate of the total velocity difference of the two clouds to within ±54 km s-1. The absolute proper-motion results are consistent with the Clouds' orbits being marginally bound to the Milky Way, albeit on an elongated orbit. The inferred relative velocity between the Clouds places them near their binding energy limit and, thus, no definitive conclusion can be made as to whether or not the Clouds are bound to one another.

The PMA Catalogue: 420 million positions and absolute proper motions

NASA Astrophysics Data System (ADS)

Akhmetov, V. S.; Fedorov, P. N.; Velichko, A. B.; Shulga, V. M.

2017-07-01

We present a catalogue that contains about 420 million absolute proper motions of stars. It was derived from the combination of positions from Gaia DR1 and 2MASS, with a mean difference of epochs of about 15 yr. Most of the systematic zonal errors inherent in the 2MASS Catalogue were eliminated before deriving the absolute proper motions. The absolute calibration procedure (zero-pointing of the proper motions) was carried out using about 1.6 million positions of extragalactic sources. The mean formal error of the absolute calibration is less than 0.35 mas yr-1. The derived proper motions cover the whole celestial sphere without gaps for a range of stellar magnitudes from 8 to 21 mag. In the sky areas where the extragalactic sources are invisible (the avoidance zone), a dedicated procedure was used that transforms the relative proper motions into absolute ones. The rms error of proper motions depends on stellar magnitude and ranges from 2-5 mas yr-1 for stars with 10 mag < G < 17 mag to 5-10 mas yr-1 for faint ones. The present catalogue contains the Gaia DR1 positions of stars for the J2015 epoch. The system of the PMA proper motions does not depend on the systematic errors of the 2MASS positions, and in the range from 14 to 21 mag represents an independent realization of a quasi-inertial reference frame in the optical and near-infrared wavelength range. The Catalogue also contains stellar magnitudes taken from the Gaia DR1 and 2MASS catalogues. A comparison of the PMA proper motions of stars with similar data from certain recent catalogues has been undertaken.

Sound Assessment through Proper Policy

ERIC Educational Resources Information Center

Chappuis, Stephen J.

2007-01-01

Aligning a school board policy manual with the faculty handbook would be an excellent application of systems thinking in support of school district mission and goals. This article talks about changing sound assessment practice in accordance with the school's proper policy. One obstacle to changing assessment practice is the prevailing belief that…

Proper Installation Improves Carpet Life.

ERIC Educational Resources Information Center

Grogan, Ralph

1998-01-01

Explains how proper carpet installation can add to carpet life; includes tips to consider before signing a carpet-installation purchasing agreement that can make the new carpet a better investment. Topics cover how color selection lengthens appearance life, the need for moisture testing, the importance of carpet seams in the purchasing process,…

PropeR revisited.

PubMed

van der Linden, Helma; Talmon, Jan; Tange, Huibert; Grimson, Jane; Hasman, Arie

2005-03-01

The PropeR EHR system (PropeRWeb) is a multidisciplinary electronic health record (EHR) system for multidisciplinary use in extramural patient care for stroke patients. The system is built using existing open source components and is based on open standards. It is implemented as a web application using servlets and Java Server Pages (JSP's) with a CORBA connection to the database servers, which are based on the OMG HDTF specifications. PropeRWeb is a generic system which can be readily customized for use in a variety of clinical domains. The system proved to be stable and flexible, although some aspects (a.o. user friendliness) could be improved. These improvements are currently under development in a second version.

16 CFR 682.3 - Proper disposal of consumer information.

Code of Federal Regulations, 2010 CFR

2010-01-01

... OF CONSUMER REPORT INFORMATION AND RECORDS § 682.3 Proper disposal of consumer information. (a) Standard. Any person who maintains or otherwise possesses consumer information for a business purpose must... 16 Commercial Practices 1 2010-01-01 2010-01-01 false Proper disposal of consumer information. 682...

16 CFR 682.3 - Proper disposal of consumer information.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 16 Commercial Practices 1 2011-01-01 2011-01-01 false Proper disposal of consumer information. 682... OF CONSUMER REPORT INFORMATION AND RECORDS § 682.3 Proper disposal of consumer information. (a) Standard. Any person who maintains or otherwise possesses consumer information for a business purpose must...

Genetic risks and healthy choices: creating citizen-consumers of genetic services through empowerment and facilitation.

PubMed

Harvey, Alison

2010-03-01

Genetic testing to identify susceptibility to a variety of common complex diseases is increasingly becoming available. In this article, focusing on the development of genetic susceptibility testing for diet-related disease, I examine the emergence of direct-to-the-consumer genetic testing services and the (re)configuration of healthcare provision, both within and outside the specialist genetics service, in the UK. I identify two key techniques within these practices: empowerment and facilitation. Using Foucauldian social theory, I show that empowerment and facilitation are being positioned as tools for the creation of citizen-consumers who will make appropriate dietary choices, based on the results of their genetic analysis. Through these techniques, individuals are transformed into properly entrepreneurial citizens who will, through judicious choices, act to maximise their 'vital capital' (their health) and the capital of the social body. I argue that the user of these services is not purely an economic figure, making rational choices as a consumer, but that her configuration as a citizen-consumer who avails herself of genetic information and services in a proper manner ensures that she is fit to contribute to the economic life of our present.

Widespread Gene Conversion in Centromere Cores

PubMed Central

Shi, Jinghua; Wolf, Sarah E.; Burke, John M.; Presting, Gernot G.; Ross-Ibarra, Jeffrey; Dawe, R. Kelly

2010-01-01

Centromeres are the most dynamic regions of the genome, yet they are typified by little or no crossing over, making it difficult to explain the origin of this diversity. To address this question, we developed a novel CENH3 ChIP display method that maps kinetochore footprints over transposon-rich areas of centromere cores. A high level of polymorphism made it possible to map a total of 238 within-centromere markers using maize recombinant inbred lines. Over half of the markers were shown to interact directly with kinetochores (CENH3) by chromatin immunoprecipitation. Although classical crossing over is fully suppressed across CENH3 domains, two gene conversion events (i.e., non-crossover marker exchanges) were identified in a mapping population. A population genetic analysis of 53 diverse inbreds suggests that historical gene conversion is widespread in maize centromeres, occurring at a rate >1×10−5/marker/generation. We conclude that gene conversion accelerates centromere evolution by facilitating sequence exchange among chromosomes. PMID:20231874

49 CFR 236.526 - Roadway element not functioning properly.

Code of Federal Regulations, 2012 CFR

2012-10-01

... 49 Transportation 4 2012-10-01 2012-10-01 false Roadway element not functioning properly. 236.526... element not functioning properly. When a roadway element except track circuit of automatic train stop... roadway element shall be caused manually to display its most restrictive aspect until such element has...

49 CFR 236.526 - Roadway element not functioning properly.

Code of Federal Regulations, 2013 CFR

2013-10-01

... 49 Transportation 4 2013-10-01 2013-10-01 false Roadway element not functioning properly. 236.526... element not functioning properly. When a roadway element except track circuit of automatic train stop... roadway element shall be caused manually to display its most restrictive aspect until such element has...

49 CFR 236.526 - Roadway element not functioning properly.

Code of Federal Regulations, 2014 CFR

2014-10-01

... 49 Transportation 4 2014-10-01 2014-10-01 false Roadway element not functioning properly. 236.526... element not functioning properly. When a roadway element except track circuit of automatic train stop... roadway element shall be caused manually to display its most restrictive aspect until such element has...

33 CFR 25.503 - Proper claimants.

Code of Federal Regulations, 2010 CFR

2010-07-01

... been an inhabitant of a foreign country at the time of the incident giving rise to the claim and must... country on a permanent basis may qualify as a proper claimant although organized under United States law...

33 CFR 25.503 - Proper claimants.

Code of Federal Regulations, 2011 CFR

2011-07-01

... been an inhabitant of a foreign country at the time of the incident giving rise to the claim and must... country on a permanent basis may qualify as a proper claimant although organized under United States law...

Perspectives of Latina and Non-Latina White Women on Barriers and Facilitators to Exercise in Pregnancy

PubMed Central

Marquez, David X.; Bustamante, Eduardo E.; Bock, Beth C.; Markenson, Glenn; Tovar, Alison; Chasan-Taber, Lisa

2009-01-01

Exercise during pregnancy has been associated with reduced risk of gestational diabetes mellitus (GDM);, however, twice as many women are sedentary during pregnancy as compared to when they are not pregnant. We conducted 3 focus groups among 20 pregnant Latina and non-Latina white women to identify barriers and facilitators to exercise in pregnancy to inform a GDM intervention study. Quantitative analyses of demographic data, and qualitative analyses of focus groups were conducted. Women identified physical limitations and restrictions, lack of resources, energy, and time as powerful exercise barriers. Social support, access to resources, information, proper diet, scheduling and the weather were identified as powerful facilitators. Intervention programs designed for pregnant women should facilitate social support, provide information and resources, as well as promote short-term and long-term benefits. PMID:20013518

Educating Children to Proper Eating Habits in the Classroom.

ERIC Educational Resources Information Center

King, Marian

A brief discussion of proper nutrition in general precedes an examination of proper nutrition for school children and the specification of nutrition education objectives for kindergarten or first grade students. The remainder of the paper delineates food projects by which objectives can be realized (for example, snack necklace, jack-o-lantern…

5 CFR 2635.205 - Proper disposition of prohibited gifts.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 5 Administrative Personnel 3 2010-01-01 2010-01-01 false Proper disposition of prohibited gifts... STANDARDS OF ETHICAL CONDUCT FOR EMPLOYEES OF THE EXECUTIVE BRANCH Gifts From Outside Sources § 2635.205 Proper disposition of prohibited gifts. (a) An employee who has received a gift that cannot be accepted...

CENP-W Plays a Role in Maintaining Bipolar Spindle Structure

PubMed Central

Kaczmarczyk, Agnieszka; Sullivan, Kevin F.

2014-01-01

The CENP-W/T complex was previously reported to be required for mitosis. HeLa cells depleted of CENP-W displayed profound mitotic defects, with mitotic timing delay, disorganized prometaphases and multipolar spindles as major phenotypic consequences. In this study, we examined the process of multipolar spindle formation induced by CENP-W depletion. Depletion of CENP-W in HeLa cells labeled with histone H2B and tubulin fluorescent proteins induced rapid fragmentation of originally bipolar spindles in a high proportion of cells. CENP-W depletion was associated with depletion of Hec1 at kinetochores. The possibility of promiscuous centrosomal duplication was ruled out by immunofluorescent examination of centrioles. However, centrioles were frequently observed to be abnormally split. In addition, a large proportion of the supernumerary poles lacked centrioles, but were positively stained with different centrosomal markers. These observations suggested that perturbation in spindle force distribution caused by defective kinetochores could contribute to a mechanical mechanism for spindle pole disruption. ‘Spindle free’ nocodazole arrested cells did not exhibit pole fragmentation after CENP-W depletion, showing that pole fragmentation is microtubule dependent. Inhibition of centrosome separation by monastrol reduced the incidence of spindle pole fragmentation, indicating that Eg5 plays a role in spindle pole disruption. Surprisingly, CENP-W depletion rescued the monopolar spindle phenotype of monastrol treatment, with an increased frequency of bipolar spindles observed after CENP-W RNAi. We overexpressed the microtubule cross-linking protein TPX2 to create spindle poles stabilized by the microtubule cross-linking activity of TPX2. Spindle pole fragmentation was suppressed in a TPX2-dependent fashion. We propose that CENP-W, by influencing proper kinetochore assembly, particularly microtubule docking sites, can confer spindle pole resistance to traction forces exerted

Second Language Listening and Unfamiliar Proper Names: Comprehension Barrier?

ERIC Educational Resources Information Center

Kobeleva, Polina P.

2012-01-01

This study examines whether unfamiliar proper names affect English as a second language (ESL) learners' listening comprehension. A total of 110 intermediate to advanced ESL learners participated; comprehension of a short news text was tested under two conditions, Names Known (all proper names pre-taught in advance) and Names Unknown (all proper…

Research on orbit prediction for solar-based calibration proper satellite

NASA Astrophysics Data System (ADS)

Chen, Xuan; Qi, Wenwen; Xu, Peng

2018-03-01

Utilizing the mathematical model of the orbit mechanics, the orbit prediction is to forecast the space target's orbit information of a certain time based on the orbit of the initial moment. The proper satellite radiometric calibration and calibration orbit prediction process are introduced briefly. On the basis of the research of the calibration space position design method and the radiative transfer model, an orbit prediction method for proper satellite radiometric calibration is proposed to select the appropriate calibration arc for the remote sensor and to predict the orbit information of the proper satellite and the remote sensor. By analyzing the orbit constraint of the proper satellite calibration, the GF-1solar synchronous orbit is chose as the proper satellite orbit in order to simulate the calibration visible durance for different satellites to be calibrated. The results of simulation and analysis provide the basis for the improvement of the radiometric calibration accuracy of the satellite remote sensor, which lays the foundation for the high precision and high frequency radiometric calibration.

Globular Clusters: Absolute Proper Motions and Galactic Orbits

NASA Astrophysics Data System (ADS)

Chemel, A. A.; Glushkova, E. V.; Dambis, A. K.; Rastorguev, A. S.; Yalyalieva, L. N.; Klinichev, A. D.

2018-04-01

We cross-match objects from several different astronomical catalogs to determine the absolute proper motions of stars within the 30-arcmin radius fields of 115 Milky-Way globular clusters with the accuracy of 1-2 mas yr-1. The proper motions are based on positional data recovered from the USNO-B1, 2MASS, URAT1, ALLWISE, UCAC5, and Gaia DR1 surveys with up to ten positions spanning an epoch difference of up to about 65 years, and reduced to Gaia DR1 TGAS frame using UCAC5 as the reference catalog. Cluster members are photometrically identified by selecting horizontal- and red-giant branch stars on color-magnitude diagrams, and the mean absolute proper motions of the clusters with a typical formal error of about 0.4 mas yr-1 are computed by averaging the proper motions of selected members. The inferred absolute proper motions of clusters are combined with available radial-velocity data and heliocentric distance estimates to compute the cluster orbits in terms of the Galactic potential models based on Miyamoto and Nagai disk, Hernquist spheroid, and modified isothermal dark-matter halo (axisymmetric model without a bar) and the same model + rotating Ferre's bar (non-axisymmetric). Five distant clusters have higher-than-escape velocities, most likely due to large errors of computed transversal velocities, whereas the computed orbits of all other clusters remain bound to the Galaxy. Unlike previously published results, we find the bar to affect substantially the orbits of most of the clusters, even those at large Galactocentric distances, bringing appreciable chaotization, especially in the portions of the orbits close to the Galactic center, and stretching out the orbits of some of the thick-disk clusters.

A comparison between families obtained from different proper elements

NASA Technical Reports Server (NTRS)

Zappala, Vincenzo; Cellino, Alberto; Farinella, Paolo

1992-01-01

Using the hierarchical method of family identification developed by Zappala et al., the results coming from the data set of proper elements computed by Williams (about 2100 numbered + about 1200 PLS 2 asteroids) and by Milani and Knezevic (5.7 version, about 4200 asteroids) are compared. Apart from some expected discrepancies due to the different data sets and/or low accuracy of proper elements computed in peculiar dynamical zones, a good agreement was found in several cases. It follows that these high reliability families represent a sample which can be considered independent on the methods used for their proper elements computation. Therefore, they should be considered as the best candidates for detailed physical studies.

[Microorganism test systems and antibiograms useful for the proper use of antibacterial agents].

PubMed

Takahashi, Shunji

2010-07-01

Antimicrobial agents are used for the accurate diagnosis of infectious diseases and effective implementation of antibacterial chemotherapy. The role of microbiological technologists is to provide data from microorganism tests useful for rapid infection treatment. Gram strain can be used to observe microorganisms and neutrophils from specimens of a patient. It is also possible to estimate the kinds of microorganism. If bacterial infectious disease is negative, there is no need for antibacterial chemotherapy. The applied dose of antibacterial agents is different in every hospital. Also, there is a difference in the percentage antibacterial agent susceptibility of isolates. Antibiograms must be created to investigate local factors. For empiric therapy, antibiograms are useful when choosing antibacterial agents showing marked efficacy against the clinical isolate. Microorganism test systems which are useful for the proper use of antibacterial agents are necessary to facilitate safe antibacterial chemotherapy and prevent the development of resistant bacteria. We report a microorganism test system employed at the Sapporo City General Hospital.

Corticotomy facilitated orthodontics: Review of a technique

PubMed Central

AlGhamdi, Ali Saad Thafeed

2009-01-01

Corticotomy found to be effective in accelerating orthodontic treatment. The most important factors in the success of this technique is proper case selection and careful surgical and orthodontic treatment. Corticotomy facilitated orthodontics advocated for comprehensive fixed orthodontic appliances in conjunction with full thickness flaps and labial and lingual corticotomies around teeth to be moved. Bone graft should be applied directly over the bone cuts and the flap sutured in place. Tooth movement should be initiated two weeks after the surgery, and every two weeks thereafter by activation of the orthodontic appliance. Orthodontic treatment time with this technique will be reduced to one-third the time of conventional orthodontics. Alveolar augmentation of labial and lingual cortical plates were used in an effort to enhance and strengthen the periodontium, reasoning that the addition of bone to alveolar housing of the teeth, using modern bone grafting techniques, ensures root coverage as the dental arch expanded. Corticotomy facilitated orthodontics is promising procedure but only few cases were reported in the literature. Controlled clinical and histological studies are needed to understand the biology of tooth movement with this procedure, the effect on teeth and bone, post-retention stability, measuring the volume of mature bone formation, and determining the status of the periodontium and roots after treatment. PMID:23960473

Facilitating Facilitators: Enhancing PBL through a Structured Facilitator Development Program

ERIC Educational Resources Information Center

Salinitri, Francine D.; Wilhelm, Sheila M.; Crabtree, Brian L.

2015-01-01

With increasing adoption of the problem-based learning (PBL) model, creative approaches to enhancing facilitator training and optimizing resources to maintain effective learning in small groups is essential. We describe a theoretical framework for the development of a PBL facilitator training program that uses the constructivist approach as the…

A question of style: method, integrity and the meaning of proper science.

PubMed

Penders, Bart; Vos, Rein; Horstman, Klasien

2009-09-01

Controversies in science often centre on methodology and integrity; these are the gatekeepers of proper science. But what exactly defines proper science as proper is not universal, especially in collaborative fields of enquiry where different perspectives meet. These encounters often result in friction, yet may equally give rise to new perspectives on proper science. A tour of collaborative endeavours like structural biology and nutrigenomics highlights some of those frictions and the controversies that can result from the encounter of 'wet' and 'dry' science. Attuning scientists to the plurality of proper science may safeguard many a scientist's integrity from being questioned unjustly.

Strategy Guideline. Proper Water Heater Selection

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hoeschele, M.; Springer, D.; German, A.

2015-04-09

This Strategy Guideline on proper water heater selection was developed by the Building America team Alliance for Residential Building Innovation to provide step-by-step procedures for evaluating preferred cost-effective options for energy efficient water heater alternatives based on local utility rates, climate, and anticipated loads.

Strategy Guideline: Proper Water Heater Selection

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hoeschele, M.; Springer, D.; German, A.

2015-04-01

This Strategy Guideline on proper water heater selection was developed by the Building America team Alliance for Residential Building Innovation to provide step-by-step procedures for evaluating preferred cost-effective options for energy efficient water heater alternatives based on local utility rates, climate, and anticipated loads.

Selecting the proper seed source

Treesearch

Robert Z. Callaham

1959-01-01

A forester faces many problems in selecting the proper seed source of ponderosa pine. He wants a seed source well adapted to all of the conditions of his planting site–one that can tolerate all of the extremes of heat, cold, and drought; that can resist the ravages of insects, diseases, and animals; that can produce the most of the desired product in the shortest time...

Proper Conformal Killing Vectors in Kantowski-Sachs Metric

NASA Astrophysics Data System (ADS)

Hussain, Tahir; Farhan, Muhammad

2018-04-01

This paper deals with the existence of proper conformal Killing vectors (CKVs) in Kantowski-Sachs metric. Subject to some integrability conditions, the general form of vector filed generating CKVs and the conformal factor is presented. The integrability conditions are solved generally as well as in some particular cases to show that the non-conformally flat Kantowski-Sachs metric admits two proper CKVs, while it admits a 15-dimensional Lie algebra of CKVs in the case when it becomes conformally flat. The inheriting conformal Killing vectors (ICKVs), which map fluid lines conformally, are also investigated.

Service Learning in Undergraduate Nursing Education: Strategies to Facilitate Meaningful Reflection.

PubMed

Schmidt, Nola A; Brown, Janet M

2016-01-01

Service learning is recognized as a valuable pedagogy involving experiential learning, reflection, and reciprocal learning. Students develop critical thinking and social awareness by using the crucial activity of reflecting upon their experiential learning with community partners. The purpose of this paper is to demystify the process of reflection by identifying best practices to enhance reflection and offering suggestions for grading. By understanding "the what" and "the how" of reflection, educators can implement service learning experiences designed to include the essential component of reflection. Strategies for facilitating meaningful reflection are described including descriptions of what students should reflect upon and how to initiate reflection through writing, reading, doing, and telling. Grading rubrics are suggested to facilitate evaluation of student reflection. When properly implemented, service learning encourages students to be good citizens of the world. By using best practices associated with reflection, students can be challenged to think critically about the world and how their service can achieve community goals. Copyright © 2016. Published by Elsevier Inc.

Fundamental Principles of Proper Space Kinematics

NASA Astrophysics Data System (ADS)

Wade, Sean

It is desirable to understand the movement of both matter and energy in the universe based upon fundamental principles of space and time. Time dilation and length contraction are features of Special Relativity derived from the observed constancy of the speed of light. Quantum Mechanics asserts that motion in the universe is probabilistic and not deterministic. While the practicality of these dissimilar theories is well established through widespread application inconsistencies in their marriage persist, marring their utility, and preventing their full expression. After identifying an error in perspective the current theories are tested by modifying logical assumptions to eliminate paradoxical contradictions. Analysis of simultaneous frames of reference leads to a new formulation of space and time that predicts the motion of both kinds of particles. Proper Space is a real, three-dimensional space clocked by proper time that is undergoing a densification at the rate of c. Coordinate transformations to a familiar object space and a mathematical stationary space clarify the counterintuitive aspects of Special Relativity. These symmetries demonstrate that within the local universe stationary observers are a forbidden frame of reference; all is in motion. In lieu of Quantum Mechanics and Uncertainty the use of the imaginary number i is restricted for application to the labeling of mass as either material or immaterial. This material phase difference accounts for both the perceived constant velocity of light and its apparent statistical nature. The application of Proper Space Kinematics will advance more accurate representations of microscopic, oscopic, and cosmological processes and serve as a foundation for further study and reflection thereafter leading to greater insight.

Fixing the reference frame for PPMXL proper motions using extragalactic sources

DOE PAGES

Grabowski, Kathleen; Carlin, Jeffrey L.; Newberg, Heidi Jo; ...

2015-05-27

In this study, we quantify and correct systematic errors in PPMXL proper motions using extragalactic sources from the first two LAMOST data releases and the Vèron-Cetty & Vèron Catalog of Quasars. Although the majority of the sources are from the Vèron catalog, LAMOST makes important contributions in regions that are not well-sampled by previous catalogs, particularly at low Galactic latitudes and in the south Galactic cap. We show that quasars in PPMXL have measurable and significant proper motions, which reflect the systematic zero-point offsets present in the catalog. We confirm the global proper motion shifts seen by Wu et al.,more » and additionally find smaller-scale fluctuations of the QSO-derived corrections to an absolute frame. Finally, we average the proper motions of 158 106 extragalactic objects in bins of 3° × 3° and present a table of proper motion corrections.« less

Computation of Asteroid Proper Elements on the Grid

NASA Astrophysics Data System (ADS)

Novakovic, B.; Balaz, A.; Knezevic, Z.; Potocnik, M.

2009-12-01

A procedure of gridification of the computation of asteroid proper orbital elements is described. The need to speed up the time consuming computations and make them more efficient is justified by the large increase of observational data expected from the next generation all sky surveys. We give the basic notion of proper elements and of the contemporary theories and methods used to compute them for different populations of objects. Proper elements for nearly 70,000 asteroids are derived since the beginning of use of the Grid infrastructure for the purpose. The average time for the catalogs update is significantly shortened with respect to the time needed with stand-alone workstations. We also present basics of the Grid computing, the concepts of Grid middleware and its Workload management system. The practical steps we undertook to efficiently gridify our application are described in full detail. We present the results of a comprehensive testing of the performance of different Grid sites, and offer some practical conclusions based on the benchmark results and on our experience. Finally, we propose some possibilities for the future work.

Improve Carpet Life with Proper Installation.

ERIC Educational Resources Information Center

Grogan, Ralph

1998-01-01

Provides guidance on ways to extend carpet life in educational facilities by paying attention to the details of carpeting installation. Issues to consider include choosing carpeting to meet specific usage requirements, are fiber protectorants used, is the installation properly done by qualified installers, and is moisture testing done prior to…

Is work keeping us from acting healthy? How workplace barriers and facilitators impact nutrition and exercise behaviors.

PubMed

Mazzola, Joseph J; Moore, J Taylor; Alexander, Katherine

2017-12-01

The purpose of this study was to identify common barriers and facilitators to healthy nutrition and exercise behaviors in the workplace and examine their relationships to those actual daily health behaviors. We utilized a concurrent embedded mixed methods approach to collect data from 93 participants over the span of four days. Participants reported 2.80 nutrition and 3.28 exercise barriers on average over the 4 days, while reporting 2.93 nutrition and 1.98 exercise facilitators in the same timeframe. Results indicated that workload and temptations around the office prevented nutritious eating; exercise behaviors were frequently hindered by workload. The most commonly mentioned eating facilitator was proper planning, while having time to exercise facilitated physical activity. Furthermore, the number of barriers reported negatively related to their respective health behaviors (i.e., more nutrition barriers translated to poorer nutrition habits) and facilitators were positively related to them, both overall and more so on the specific day they were reported. The implications of these finding show the importance of barriers/facilitators in the workplace and aid in the creation of more targeted health promotion that could increase positive employee health behaviors by eliminating common barriers and enhancing facilitators. Copyright © 2016 John Wiley & Sons, Ltd.

An All-Sky Search for Wide Binaries in the SUPERBLINK Proper Motion Catalog

NASA Astrophysics Data System (ADS)

Hartman, Zachary; Lepine, Sebastien

2017-01-01

We present initial results from an all-sky search for Common Proper Motion (CPM) binaries in the SUPERBLINK all-sky proper motion catalog of 2.8 million stars with proper motions greater than 40 mas/yr, which has been recently enhanced with data from the GAIA mission. We initially search the SUPERBLINK catalog for pairs of stars with angular separations up to 1 degree and proper motion difference less than 40 mas/yr. In order to determine which of these pairs are real binaries, we develop a Bayesian analysis to calculate probabilities of true companionship based on a combination of proper motion magnitude, angular separation, and proper motion differences. The analysis reveals that the SUPERBLINK catalog most likely contains ~40,000 genuine common proper motion binaries. We provide initial estimates of the distances and projected physical separations of these wide binaries.

21 CFR 610.62 - Proper name; package label; legible type.

Code of Federal Regulations, 2010 CFR

2010-04-01

... (CONTINUED) BIOLOGICS GENERAL BIOLOGICAL PRODUCTS STANDARDS Labeling Standards § 610.62 Proper name; package... contrast in color value between the proper name and the background shall be at least as great as the color value between the trademark and trade name and the background. Typography, layout, contrast, and other...

Participation in "Handwashing University" Promotes Proper Handwashing Techniques for Youth

ERIC Educational Resources Information Center

Fenton, Ginger; Radhakrishna, Rama; Cutter, Catherine Nettles

2010-01-01

A study was conducted to assess the effectiveness of the Handwashing University on teaching youth the benefits of proper handwashing. The Handwashing University is an interactive display with several successive stations through which participants move to learn necessary skills for proper handwashing. Upon completion of the Handwashing University,…

Measuring mitotic forces.

PubMed

Ye, Anna A; Maresca, Thomas J

2018-01-01

Productive chromosome movements require that a large multiprotein complex called the kinetochore assemble on sister centromeres. The kinetochore fulfills two critical functions as (1) the physical linkage between chromosomes and spindle microtubules and (2) a mechanomolecular sensor that relays a spindle assembly checkpoint signal delaying anaphase onset until chromosomes are attached to spindle microtubules and bioriented. Given its central roles in such a vital process, the kinetochore is one of the most important force-transducing structures in cells; yet it has been technically challenging to measure kinetochore forces. Barriers to measuring cellular forces have begun to be broken by the development of fluorescence-based tension sensors. In this chapter, two methods will be described for measuring kinetochore forces in living cells and strategies for applying these sensors to other force-transducing processes and molecules will be discussed. © 2018 Elsevier Inc. All rights reserved.

20 CFR 658.415 - Transferring complaints to proper JS office.

Code of Federal Regulations, 2010 CFR

2010-04-01

... 20 Employees' Benefits 3 2010-04-01 2010-04-01 false Transferring complaints to proper JS office... Agency Js Complaint System § 658.415 Transferring complaints to proper JS office. (a) Where a JS-related... local office serving the area in which the employer is located. Where a JS-related complaint deals with...

Activity of a C-terminal plant homeodomain (PHD) of Msc1 is essential for function.

PubMed

Qiu, Xinxing; Dul, Barbara E; Walworth, Nancy C

2010-11-19

Msc1, a member of the Jarid1 family of putative histone demethylases, is required for chromosome stability in fission yeast. Msc1 associates with the Swr1 complex that facilitates deposition of histone H2A.Z into chromatin. To assess the function of Msc1 in the Swr1 complex, domains of Msc1 necessary for interaction with Swr1 were identified. The C-terminal plant homeodomain (PHD) 2 and PHD3 of Msc1 are sufficient to confer association with Swr1 and allow Msc1 to function in the context of kinetochore mutants. On the other hand, a mutant with a single amino acid substitution in PHD2 within the full-length Msc1 protein retains the ability to bind to Swr1 but eliminates the function of Msc1 in combination with kinetochore mutants. Thus, Swr1 association is critical but not sufficient for Msc1 function. An activity of Msc1 that depends on the cysteine residue within PHD2 of Msc1 is likewise critical for function. On the basis of our observation that the PHDs of Msc1 act as E3 ubiquitin ligases and that mutations of cysteine residues within those domains abolish ligase activity, we speculate that the ability of Msc1 to facilitate ubiquitin transfer is critical for the function it mediates through its association with Swr1.

Activity of a C-terminal Plant Homeodomain (PHD) of Msc1 Is Essential for Function*

PubMed Central

Qiu, Xinxing; Dul, Barbara E.; Walworth, Nancy C.

2010-01-01

Msc1, a member of the Jarid1 family of putative histone demethylases, is required for chromosome stability in fission yeast. Msc1 associates with the Swr1 complex that facilitates deposition of histone H2A.Z into chromatin. To assess the function of Msc1 in the Swr1 complex, domains of Msc1 necessary for interaction with Swr1 were identified. The C-terminal plant homeodomain (PHD) 2 and PHD3 of Msc1 are sufficient to confer association with Swr1 and allow Msc1 to function in the context of kinetochore mutants. On the other hand, a mutant with a single amino acid substitution in PHD2 within the full-length Msc1 protein retains the ability to bind to Swr1 but eliminates the function of Msc1 in combination with kinetochore mutants. Thus, Swr1 association is critical but not sufficient for Msc1 function. An activity of Msc1 that depends on the cysteine residue within PHD2 of Msc1 is likewise critical for function. On the basis of our observation that the PHDs of Msc1 act as E3 ubiquitin ligases and that mutations of cysteine residues within those domains abolish ligase activity, we speculate that the ability of Msc1 to facilitate ubiquitin transfer is critical for the function it mediates through its association with Swr1. PMID:20858896

16 CFR 23.15 - Misuse of the term “properly cut,” etc.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 16 Commercial Practices 1 2011-01-01 2011-01-01 false Misuse of the term âproperly cut,â etc. 23... FOR THE JEWELRY, PRECIOUS METALS, AND PEWTER INDUSTRIES § 23.15 Misuse of the term “properly cut,” etc. It is unfair or deceptive to use the terms “properly cut,” “proper cut,” “modern cut,” or any...

16 CFR 23.15 - Misuse of the term “properly cut,” etc.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 16 Commercial Practices 1 2010-01-01 2010-01-01 false Misuse of the term âproperly cut,â etc. 23... FOR THE JEWELRY, PRECIOUS METALS, AND PEWTER INDUSTRIES § 23.15 Misuse of the term “properly cut,” etc. It is unfair or deceptive to use the terms “properly cut,” “proper cut,” “modern cut,” or any...

Secular Extragalactic Parallax and Geometric Distances with Gaia Proper Motions

NASA Astrophysics Data System (ADS)

Paine, Jennie; Darling, Jeremiah K.

2018-06-01

The motion of the Solar System with respect to the cosmic microwave background (CMB) rest frame creates a well measured dipole in the CMB, which corresponds to a linear solar velocity of about 78 AU/yr. This motion causes relatively nearby extragalactic objects to appear to move compared to more distant objects, an effect that can be measured in the proper motions of nearby galaxies. An object at 1 Mpc and perpendicular to the CMB apex will exhibit a secular parallax, observed as a proper motion, of 78 µas/yr. The relatively large peculiar motions of galaxies make the detection of secular parallax challenging for individual objects. Instead, a statistical parallax measurement can be made for a sample of objects with proper motions, where the global parallax signal is modeled as an E-mode dipole that diminishes linearly with distance. We present preliminary results of applying this model to a sample of nearby galaxies with Gaia proper motions to detect the statistical secular parallax signal. The statistical measurement can be used to calibrate the canonical cosmological “distance ladder.”

Boltzmann babies in the proper time measure

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bousso, Raphael; Freivogel, Ben; Yang, I-S.

2008-05-15

After commenting briefly on the role of the typicality assumption in science, we advocate a phenomenological approach to the cosmological measure problem. Like any other theory, a measure should be simple, general, well defined, and consistent with observation. This allows us to proceed by elimination. As an example, we consider the proper time cutoff on a geodesic congruence. It predicts that typical observers are quantum fluctuations in the early universe, or Boltzmann babies. We sharpen this well-known youngness problem by taking into account the expansion and open spatial geometry of pocket universes. Moreover, we relate the youngness problem directly tomore » the probability distribution for observables, such as the temperature of the cosmic background radiation. We consider a number of modifications of the proper time measure, but find none that would make it compatible with observation.« less

Boltzmann babies in the proper time measure

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bousso, Raphael; Bousso, Raphael; Freivogel, Ben

After commenting briefly on the role of the typicality assumption in science, we advocate a phenomenological approach to the cosmological measure problem. Like any other theory, a measure should be simple, general, well defined, and consistent with observation. This allows us to proceed by elimination. As an example, we consider the proper time cutoff on a geodesic congruence. It predicts that typical observers are quantum fluctuations in the early universe, or Boltzmann babies. We sharpen this well-known youngness problem by taking into account the expansion and open spatial geometry of pocket universes. Moreover, we relate the youngness problem directly tomore » the probability distribution for observables, such as the temperature of the cosmic background radiation. We consider a number of modifications of the proper time measure, but find none that would make it compatible with observation.« less

21 CFR 610.62 - Proper name; package label; legible type.

Code of Federal Regulations, 2011 CFR

2011-04-01

... 21 Food and Drugs 7 2011-04-01 2010-04-01 true Proper name; package label; legible type. 610.62 Section 610.62 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES... contrast in color value between the proper name and the background shall be at least as great as the color...

De novo pathogenic variants in CHAMP1 are associated with global developmental delay, intellectual disability, and dysmorphic facial features.

PubMed

Tanaka, Akemi J; Cho, Megan T; Retterer, Kyle; Jones, Julie R; Nowak, Catherine; Douglas, Jessica; Jiang, Yong-Hui; McConkie-Rosell, Allyn; Schaefer, G Bradley; Kaylor, Julie; Rahman, Omar A; Telegrafi, Aida; Friedman, Bethany; Douglas, Ganka; Monaghan, Kristin G; Chung, Wendy K

2016-01-01

We identified five unrelated individuals with significant global developmental delay and intellectual disability (ID), dysmorphic facial features and frequent microcephaly, and de novo predicted loss-of-function variants in chromosome alignment maintaining phosphoprotein 1 (CHAMP1). Our findings are consistent with recently reported de novo mutations in CHAMP1 in five other individuals with similar features. CHAMP1 is a zinc finger protein involved in kinetochore-microtubule attachment and is required for regulating the proper alignment of chromosomes during metaphase in mitosis. Mutations in CHAMP1 may affect cell division and hence brain development and function, resulting in developmental delay and ID.

GIP Contributions to the Regulation of Centromere at the Interface Between the Nuclear Envelope and the Nucleoplasm.

PubMed

Chabouté, Marie-Edith; Berr, Alexandre

2016-01-01

Centromeres are known as specific chromatin domains without which eukaryotic cells cannot divide properly during mitosis. Despite the considerable efforts to understand the centromere/kinetochore assembly during mitosis, until recently, comparatively few studies have dealt with the regulation of centromere during interphase. Here, we briefly review and discuss past and recent advances about the architecture of centromeres and their regulation during the cell cycle. Furthermore, we highlight and discuss new findings and hypotheses regarding the specific regulation of centromeres in both plant and animal nuclei, especially with GIP proteins at the interface between the nuclear envelope and the nucleoplasm.

The Essentials of Proper Wine Service.

ERIC Educational Resources Information Center

Manago, Gary H.

This instructional unit was designed to assist the food services instructor and/or the restaurant manager in training students and/or staff in the proper procedure for serving wines to guests. The lesson plans included in this unit focus on: (1) the different types of wine glasses and their uses; (2) the parts of a wine glass; (3) the proper…

N-terminal regions of Mps1 kinase determine functional bifurcation.

PubMed

Araki, Yasuhiro; Gombos, Linda; Migueleti, Suellen P S; Sivashanmugam, Lavanya; Antony, Claude; Schiebel, Elmar

2010-04-05

Mps1 is a conserved kinase that in budding yeast functions in duplication of the spindle pole body (SPB), spindle checkpoint activation, and kinetochore biorientation. The identity of Mps1 targets and the subdomains that convey specificity remain largely unexplored. Using a novel combination of systematic deletion analysis and chemical biology, we identified two regions within the N terminus of Mps1 that are essential for either SPB duplication or kinetochore biorientation. Suppression analysis of the MPS1 mutants defective in SPB duplication and biochemical enrichment of Mps1 identified the essential SPB components Spc29 and the yeast centrin Cdc31 as Mps1 targets in SPB duplication. Our data suggest that phosphorylation of Spc29 by Mps1 in G1/S recruits the Mps2-Bbp1 complex to the newly formed SPB to facilitate its insertion into the nuclear envelope. Mps1 phosphorylation of Cdc31 at the conserved T110 residue controls substrate binding to Kar1 protein. These findings explain the multiple SPB duplication defects of mps1 mutants on a molecular level.

Proper shoe sizes for Thai elderly.

PubMed

Chaiwanichsiri, Dootchai; Tantisiriwat, Natthiya; Janchai, Siriporn

2008-12-01

Problems from improper shoe fitting are common, but there are limited foot data for the older Thai population. To study foot dimensions and determine proper shoe sizes for Thai elderly. Healthy older people: 108 men, 105 women, aged 60-80 years, who were independent in walking, were recruited. Thirteen foot dimensions and current shoes used were measured. Side-to-side, gender difference, and correlations of main foot measurements were analyzed. About 50% women and 34% men wore too narrow shoes, and this was found to be associated with foot pain. At the same foot length (FL), men had larger foot width (FW) and toe depth. Foot width=2.39+(0.29 x FL), r=0.50, p=0.001 for women and=2.48+(0.31 x FL), r=0.56, p=0.002 for men. Arch length=1.0+(0.7 x FL), r=0.93, p=0.001 for both genders. Toe depth had constant values in all shoe sizes of each gender. Correlations of other foot parameters were reported. These anthropometric data is essential for proper shoe fitting in order to provide foot ergonomics and prevent foot problems for older Thai people.

DNA Binding of Centromere Protein C (CENPC) Is Stabilized by Single-Stranded RNA

PubMed Central

Du, Yaqing; Topp, Christopher N.; Dawe, R. Kelly

2010-01-01

Centromeres are the attachment points between the genome and the cytoskeleton: centromeres bind to kinetochores, which in turn bind to spindles and move chromosomes. Paradoxically, the DNA sequence of centromeres has little or no role in perpetuating kinetochores. As such they are striking examples of genetic information being transmitted in a manner that is independent of DNA sequence (epigenetically). It has been found that RNA transcribed from centromeres remains bound within the kinetochore region, and this local population of RNA is thought to be part of the epigenetic marking system. Here we carried out a genetic and biochemical study of maize CENPC, a key inner kinetochore protein. We show that DNA binding is conferred by a localized region 122 amino acids long, and that the DNA-binding reaction is exquisitely sensitive to single-stranded RNA. Long, single-stranded nucleic acids strongly promote the binding of CENPC to DNA, and the types of RNAs that stabilize DNA binding match in size and character the RNAs present on kinetochores in vivo. Removal or replacement of the binding module with HIV integrase binding domain causes a partial delocalization of CENPC in vivo. The data suggest that centromeric RNA helps to recruit CENPC to the inner kinetochore by altering its DNA binding characteristics. PMID:20140237

20 CFR 602.41 - Proper expenditure of Quality Control granted funds.

Code of Federal Regulations, 2011 CFR

2011-04-01

... 20 Employees' Benefits 3 2011-04-01 2011-04-01 false Proper expenditure of Quality Control granted... LABOR QUALITY CONTROL IN THE FEDERAL-STATE UNEMPLOYMENT INSURANCE SYSTEM Quality Control Grants to States § 602.41 Proper expenditure of Quality Control granted funds. The Secretary may, after reasonable...

20 CFR 602.41 - Proper expenditure of Quality Control granted funds.

Code of Federal Regulations, 2010 CFR

2010-04-01

... 20 Employees' Benefits 3 2010-04-01 2010-04-01 false Proper expenditure of Quality Control granted... LABOR QUALITY CONTROL IN THE FEDERAL-STATE UNEMPLOYMENT INSURANCE SYSTEM Quality Control Grants to States § 602.41 Proper expenditure of Quality Control granted funds. The Secretary may, after reasonable...

Exploring Facilitators of Post-traumatic Growth in Patients with Spinal Cord Injury: A Qualitative Study

PubMed Central

Khanjani, Mohammad Saeed; Younesi, Seyed Jalal; Khankeh, Hamid Reza; Azkhosh, Manouchehr

2017-01-01

Introduction There is increasing evidence regarding people’s reactions to life stressors in which people also may show positive experiences following a traumatic event. The aim of the present study was to explain the facilitators of post-traumatic growth based on the experiences of patients with a spinal cord injury. Methods This was a qualitative study conducted on 16 Iranian patients with a spinal cord injury using semistructured, in-depth interviews, and content analysis in 2015. These participants, despite their spinal cord injury, were successful in their lives and were considered successful members of society. A purposive sampling method was used until reaching data saturation, and then the collected data were analyzed using a content analysis method. Results The study revealed several factors as facilitators of post-traumatic growth in the patients. The extracted facilitators were put into seven categories of main concepts, including existence of support resources, contact with spinal cord injury associations, spiritual beliefs, positive attitude toward injury, access to proper facilities, enhancement of knowledge and awareness, and active presence in society. Conclusion Different factors may facilitate post-traumatic growth in patients with a spinal cord injury. Understanding these facilitators may help us in designing educational, support, and consulting programs for patients and their families as well as to the correct the support programs. PMID:28243405

1 CFR 51.9 - What is the proper language of incorporation?

Code of Federal Regulations, 2010 CFR

2010-01-01

... 1 General Provisions 1 2010-01-01 2010-01-01 false What is the proper language of incorporation... What is the proper language of incorporation? (a) The language incorporating a publication by reference... is intended and completed by the final rule document in which it appears. (b) The language...

1 CFR 51.9 - What is the proper language of incorporation?

Code of Federal Regulations, 2011 CFR

2011-01-01

... 1 General Provisions 1 2011-01-01 2011-01-01 false What is the proper language of incorporation... What is the proper language of incorporation? (a) The language incorporating a publication by reference... is intended and completed by the final rule document in which it appears. (b) The language...

The UKIDSS-2MASS proper motion survey - I. Ultracool dwarfs from UKIDSS DR4

NASA Astrophysics Data System (ADS)

Deacon, N. R.; Hambly, N. C.; King, R. R.; McCaughrean, M. J.

2009-04-01

The UK Infrared Telescope Infrared Deep Sky Survey (UKIDSS) is the first of a new generation of infrared surveys. Here, we combine the data from two UKIDSS components, the Large Area Survey (LAS) and the Galactic Cluster Survey (GCS), with Two-Micron All-Sky Survey (2MASS) data to produce an infrared proper motion survey for low-mass stars and brown dwarfs. In total, we detect 267 low-mass stars and brown dwarfs with significant proper motions. We recover all 10 known single L dwarfs and the one known T dwarf above the 2MASS detection limit in our LAS survey area and identify eight additional new candidate L dwarfs. We also find one new candidate L dwarf in our GCS sample. Our sample also contains objects from 11 potential common proper motion binaries. Finally, we test our proper motions and find that while the LAS objects have proper motions consistent with absolute proper motions, the GCS stars may have proper motions which are significantly underestimated. This is possibly due to the bulk motion of some of the local astrometric reference stars used in the proper motion determination.

45 CFR 170.423 - Principles of proper conduct for ONC-ATCBs.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 45 Public Welfare 1 2010-10-01 2010-10-01 false Principles of proper conduct for ONC-ATCBs. 170... § 170.423 Principles of proper conduct for ONC-ATCBs. An ONC-ATCB shall: (a) Operate its certification... EHR Module developer conspicuously include the following text on its Web site and in all marketing...

Crutches and children - proper fit and safety tips

MedlinePlus

... 000640.htm Crutches and children - proper fit and safety tips To use the sharing features on this ... the crutch, then extended when taking a step. Safety Tips Teach your child to: Always keep crutches ...

CDK-dependent potentiation of MPS1 kinase activity is essential to the mitotic checkpoint.

PubMed

Morin, Violeta; Prieto, Susana; Melines, Sabrina; Hem, Sonia; Rossignol, Michel; Lorca, Thierry; Espeut, Julien; Morin, Nathalie; Abrieu, Ariane

2012-02-21

Accurate chromosome segregation relies upon a mitotic checkpoint that monitors kinetochore attachment toward opposite spindle poles before enabling chromosome disjunction [1]. The MPS1/TTK protein kinase is a core component of the mitotic checkpoint that lies upstream of MAD2 and BubR1 both at the kinetochore and in the cytoplasm [2, 3]. To gain insight into the mechanisms underlying the regulation of MPS1 kinase, we undertook the identification of Xenopus MPS1 phosphorylation sites by mass spectrometry. We mapped several phosphorylation sites onto MPS1 and we show that phosphorylation of S283 in the noncatalytic region of MPS1 is required for full kinase activity. This phosphorylation potentiates MPS1 catalytic efficiency without impairing its affinity for the substrates. By using Xenopus egg extracts depleted of endogenous MPS1 and reconstituted with single point mutants, we show that phosphorylation of S283 is essential to activate the mitotic checkpoint. This phosphorylation does not regulate the localization of MPS1 to the kinetochore but is required for the recruitment of MAD1/MAD2, demonstrating its role at the kinetochore. Constitutive phosphorylation of S283 lowers the number of kinetochores required to hold the checkpoint, which suggests that CDK-dependent phosphorylation of MPS1 is essential to sustain the mitotic checkpoint when few kinetochores remain unattached. Copyright © 2012 Elsevier Ltd. All rights reserved.

Practice Facilitators' and Leaders' Perspectives on a Facilitated Quality Improvement Program.

PubMed

McHugh, Megan; Brown, Tiffany; Liss, David T; Walunas, Theresa L; Persell, Stephen D

2018-04-01

Practice facilitation is a promising approach to helping practices implement quality improvements. Our purpose was to describe practice facilitators' and practice leaders' perspectives on implementation of a practice facilitator-supported quality improvement program and describe where their perspectives aligned and diverged. We conducted interviews with practice leaders and practice facilitators who participated in a program that included 35 improvement strategies aimed at the ABCS of heart health (aspirin use in high-risk individuals, blood pressure control, cholesterol management, and smoking cessation). Rapid qualitative analysis was used to collect, organize, and analyze the data. We interviewed 17 of the 33 eligible practice leaders, and the 10 practice facilitators assigned to those practices. Practice leaders and practice facilitators both reported value in the program's ability to bring needed, high-quality resources to practices. Practice leaders appreciated being able to set the schedule for facilitation and select among the 35 interventions. According to practice facilitators, however, relying on practice leaders to set the pace of the intervention resulted in a lower level of program intensity than intended. Practice leaders preferred targeted assistance, particularly electronic health record documentation guidance and linkages to state smoking cessation programs. Practice facilitators reported that the easiest interventions were those that did not alter care practices. The dual perspectives of practice leaders and practice facilitators provide a more holistic picture of enablers and barriers to program implementation. There may be greater opportunities to assist small practices through simple, targeted practice facilitator-supported efforts rather than larger, comprehensive quality improvement projects. © 2018 Annals of Family Medicine, Inc.

Assigned value improves memory of proper names.

PubMed

Festini, Sara B; Hartley, Alan A; Tauber, Sarah K; Rhodes, Matthew G

2013-01-01

Names are more difficult to remember than other personal information such as occupations. The current research examined the influence of assigned point value on memory and metamemory judgements for names and occupations to determine whether incentive can improve recall of proper names. In Experiment 1 participants studied face-name and face-occupation pairs assigned 1 or 10 points, made judgements of learning, and were given a cued recall test. High-value names were recalled more often than low-value names. However, recall of occupations was not influenced by value. In Experiment 2 meaningless nonwords were used for both names and occupations. The name difficulty disappeared, and value influenced recall of both names and occupations. Thus value similarly influenced names and occupations when meaningfulness was held constant. In Experiment 3 participants were required to use overt rote rehearsal for all items. Value did not boost recall of high-value names, suggesting that differential processing could not be implemented to improve memory. Thus incentives may improve memory for proper names by motivating people to engage in selective rehearsal and effortful elaborative processing.

Inhibition of CDK7 bypasses spindle assembly checkpoint via premature cyclin B degradation during oocyte meiosis.

PubMed

Wang, HaiYang; Jo, Yu-Jin; Sun, Tian-Yi; Namgoong, Suk; Cui, Xiang-Shun; Oh, Jeong Su; Kim, Nam-Hyung

2016-12-01

To ensure accurate chromosome segregation, the spindle assembly checkpoint (SAC) delays anaphase onset by preventing the premature activation of anaphase-promoting complex/cyclosome (APC/C) until all kinetochores are attached to the spindle. Although an escape from mitosis in the presence of unsatisfied SAC has been shown in several cancer cells, it has not been reported in oocyte meiosis. Here, we show that CDK7 activity is required to prevent a bypass of SAC during meiosis I in mouse oocytes. Inhibition of CDK7 using THZ1 accelerated the first meiosis, leading to chromosome misalignment, lag of chromosomes during chromosome segregation, and a high incidence of aneuploidy. Notably, this acceleration occurred in the presence of SAC proteins including Mad2 and Bub3 at the kinetochores. However, inhibition of APC/C-mediated cyclin B degradation blocked the THZ1-induced premature polar body extrusion. Moreover, chromosomal defects mediated by THZ1 were rescued when anaphase onset was delayed. Collectively, our results show that CDK7 activity is required to prevent premature anaphase onset by suppressing the bypass of SAC, thus ensuring chromosome alignment and proper segregation. These findings reveal new roles of CDK7 in the regulation of meiosis in mammalian oocytes. Copyright © 2016 Elsevier B.V. All rights reserved.

Using a voice to put a name to a face: the psycholinguistics of proper name comprehension.

PubMed

Barr, Dale J; Jackson, Laura; Phillips, Isobel

2014-02-01

We propose that hearing a proper name (e.g., Kevin) in a particular voice serves as a compound memory cue that directly activates representations of a mutually known target person, often permitting reference resolution without any complex computation of shared knowledge. In a referential communication study, pairs of friends played a communication game, in which we monitored the eyes of one friend (the addressee) while he or she sought to identify the target person, in a set of four photos, on the basis of a name spoken aloud. When the name was spoken by a friend, addressees rapidly identified the target person, and this facilitation was independent of whether the friend was articulating a message he or she had designed versus one from a third party with whom the target person was not shared. Our findings suggest that the comprehension system takes advantage of regularities in the environment to minimize effortful computation about who knows what.

Facilitators and Barriers to Adopting Robotic-Assisted Surgery: Contextualizing the Unified Theory of Acceptance and Use of Technology

PubMed Central

BenMessaoud, Christine; Kharrazi, Hadi; MacDorman, Karl F.

2011-01-01

Robotic-assisted surgical techniques are not yet well established among surgeon practice groups beyond a few surgical subspecialties. To help identify the facilitators and barriers to their adoption, this belief-elicitation study contextualized and supplemented constructs of the unified theory of acceptance and use of technology (UTAUT) in robotic-assisted surgery. Semi-structured individual interviews were conducted with 21 surgeons comprising two groups: users and nonusers. The main facilitators to adoption were Perceived Usefulness and Facilitating Conditions among both users and nonusers, followed by Attitude Toward Using Technology among users and Extrinsic Motivation among nonusers. The three main barriers to adoption for both users and nonusers were Perceived Ease of Use and Complexity, Perceived Usefulness, and Perceived Behavioral Control. This study's findings can assist surgeons, hospital and medical school administrators, and other policy makers on the proper adoption of robotic-assisted surgery and can guide future research on the development of theories and framing of hypotheses. PMID:21283719

Proper motion of the Draco dwarf galaxy from Subaru Suprime-Cam data

NASA Astrophysics Data System (ADS)

Casetti-Dinescu, Dana I.; Girard, Terrence M.

2016-09-01

We have measured the absolute proper motion of the Draco dwarf spheroidal galaxy using Subaru Suprime-Cam images taken at three epochs, with time baselines of 4.4 and 7 yr. The magnitude limit of the proper-motion study is I = 25, thus allowing for thousands of background galaxies and Draco stars to be used to perform extensive astrometric tests and to derive the correction to an inertial reference frame. The derived proper motion is (μα, μδ) = (-0.284 ± 0.047, -0.289 ± 0.041) mas yr-1. This motion implies an orbit that takes Draco to a pericentre of ˜20 kpc; a somewhat disruptive orbit suggesting that tides might account for the rising velocity-dispersion profile of Draco seen in line-of-sight velocity studies. The orbit is only marginally consistent with Draco's membership to the vast polar structure of Galactic satellites, in contrast to a recent Hubble Space Telescope proper-motion measurement that finds alignment very likely. Our study is a test case to demonstrate that deep imaging with mosaic cameras of appropriate resolution can be used for high-accuracy, ground-based proper-motion measurement. As a useful by-product of the study, we also identify two faint brown-dwarf candidates in the foreground field.

A new UKIDSS proper motion survey and key early results, including new benchmark systems

NASA Astrophysics Data System (ADS)

Smith, L.; Lucas, P.; Burningham, B.; Jones, H.; Pinfield, D.; Smart, R.; Andrei, A.

We present a proper motion catalogue for the 1500 deg2 of 2 epoch J-band UKIDSS Large Area Survey (LAS) data, which includes 120,000 stellar sources with motions detected above the 5sigma level. Our upper limit on proper motion detection is 3\\farcs3 yr-1 and typical uncertainties are of order 10 mas yr-1 for bright sources from data with a modest 1.8-7.0 year epoch baseline. We developed a bespoke proper motion pipeline which applies a source-unique second order polynomial transformation to UKIDSS array coordinates to counter potential local non-uniformity in the focal plane. Our catalogue agrees well with the proper motion data supplied in the current WFCAM Science Archive (WSA) tenth data release (DR10) catalogue where there is overlap, and in various optical catalogues, but it benefits from some improvements, such as a larger matching radius and relative to absolute proper motion correction. We present proper motion results for 128 T dwarfs in the UKIDSS LAS and key early results of projects utilising our catalogue, in particular searches for brown dwarf benchmark systems through cross matches with existing proper motion catalogues. We report the discovery of two new T dwarf benchmark systems.

The Influence of Facilitator and Facilitation Characteristics on Participants' Ratings of Stepfamily Education

ERIC Educational Resources Information Center

Higginbotham, Brian J.; Myler, Cory

2010-01-01

We examine the relative importance of facilitator and facilitation characteristics on participant ratings of a stepfamily education program. Data from 48 facilitators and 598 participants suggest that quality facilitation is more meaningful to participants than whether facilitators have comparable demographic characteristics or life experiences.…

Limited-memory adaptive snapshot selection for proper orthogonal decomposition

DOE Office of Scientific and Technical Information (OSTI.GOV)

Oxberry, Geoffrey M.; Kostova-Vassilevska, Tanya; Arrighi, Bill

2015-04-02

Reduced order models are useful for accelerating simulations in many-query contexts, such as optimization, uncertainty quantification, and sensitivity analysis. However, offline training of reduced order models can have prohibitively expensive memory and floating-point operation costs in high-performance computing applications, where memory per core is limited. To overcome this limitation for proper orthogonal decomposition, we propose a novel adaptive selection method for snapshots in time that limits offline training costs by selecting snapshots according an error control mechanism similar to that found in adaptive time-stepping ordinary differential equation solvers. The error estimator used in this work is related to theory boundingmore » the approximation error in time of proper orthogonal decomposition-based reduced order models, and memory usage is minimized by computing the singular value decomposition using a single-pass incremental algorithm. Results for a viscous Burgers’ test problem demonstrate convergence in the limit as the algorithm error tolerances go to zero; in this limit, the full order model is recovered to within discretization error. The resulting method can be used on supercomputers to generate proper orthogonal decomposition-based reduced order models, or as a subroutine within hyperreduction algorithms that require taking snapshots in time, or within greedy algorithms for sampling parameter space.« less

49 CFR 109.11 - Assistance of properly qualified personnel.

Code of Federal Regulations, 2013 CFR

2013-10-01

... MATERIALS SAFETY ADMINISTRATION, DEPARTMENT OF TRANSPORTATION HAZARDOUS MATERIALS AND OIL TRANSPORTATION... properly qualified to perform a function that is essential to the agent's exercise of authority under this...

Tunable modulation of refracted lamb wave front facilitated by adaptive elastic metasurfaces

NASA Astrophysics Data System (ADS)

Li, Shilong; Xu, Jiawen; Tang, J.

2018-01-01

This letter reports designs of adaptive metasurfaces capable of modulating incoming wave fronts of elastic waves through electromechanical-tuning of their cells. The proposed elastic metasurfaces are composed of arrayed piezoelectric units with individually connected negative capacitance elements that are online tunable. By adjusting the negative capacitances properly, accurately formed, discontinuous phase profiles along the elastic metasurfaces can be achieved. Subsequently, anomalous refraction with various angles can be realized on the transmitted lowest asymmetric mode Lamb wave. Moreover, designs to facilitate planar focal lenses and source illusion devices can also be accomplished. The proposed flexible and versatile strategy to manipulate elastic waves has potential applications ranging from structural fault detection to vibration/noise control.

27 CFR 4.45 - Certificates of origin, identity and proper cellar treatment.

Code of Federal Regulations, 2010 CFR

2010-04-01

..., identity and proper cellar treatment. 4.45 Section 4.45 Alcohol, Tobacco Products and Firearms ALCOHOL AND... Requirements for Withdrawal of Wine From Customs Custody § 4.45 Certificates of origin, identity and proper cellar treatment. (a) Origin and identity. Imported wine shall not be released from customs custody for...

27 CFR 4.45 - Certificates of origin, identity and proper cellar treatment.

Code of Federal Regulations, 2011 CFR

2011-04-01

..., identity and proper cellar treatment. 4.45 Section 4.45 Alcohol, Tobacco Products and Firearms ALCOHOL AND... Requirements for Withdrawal of Wine From Customs Custody § 4.45 Certificates of origin, identity and proper cellar treatment. (a) Origin and identity. Imported wine shall not be released from customs custody for...

27 CFR 4.45 - Certificates of origin, identity and proper cellar treatment.

Code of Federal Regulations, 2012 CFR

2012-04-01

..., identity and proper cellar treatment. 4.45 Section 4.45 Alcohol, Tobacco Products and Firearms ALCOHOL AND... Requirements for Withdrawal of Wine From Customs Custody § 4.45 Certificates of origin, identity and proper cellar treatment. (a) Origin and identity. Imported wine shall not be released from customs custody for...

27 CFR 4.45 - Certificates of origin, identity and proper cellar treatment.

Code of Federal Regulations, 2013 CFR

2013-04-01

..., identity and proper cellar treatment. 4.45 Section 4.45 Alcohol, Tobacco Products and Firearms ALCOHOL AND... Requirements for Withdrawal of Wine From Customs Custody § 4.45 Certificates of origin, identity and proper cellar treatment. (a) Origin and identity. Imported wine shall not be released from customs custody for...

27 CFR 4.45 - Certificates of origin, identity and proper cellar treatment.

Code of Federal Regulations, 2014 CFR

2014-04-01

..., identity and proper cellar treatment. 4.45 Section 4.45 Alcohol, Tobacco Products and Firearms ALCOHOL AND... Requirements for Withdrawal of Wine From Customs Custody § 4.45 Certificates of origin, identity and proper cellar treatment. (a) Origin and identity. Imported wine shall not be released from customs custody for...

Cresting the wave: proper motions of the Eastern Banded Structure

NASA Astrophysics Data System (ADS)

Deason, Alis J.; Belokurov, Vasily; Koposov, Sergey E.

2018-01-01

We study the kinematic properties of the Eastern Banded Structure (EBS) and Hydra I overdensity using exquisite proper motions derived from the Sloan Digital Sky Survey (SDSS) and Gaia source catalogue. Main sequence turn-off stars in the vicinity of the EBS are identified from SDSS photometry; we use the proper motions and, where applicable, spectroscopic measurements of these stars to probe the kinematics of this apparent stream. We find that the EBS and Hydra I share common kinematic and chemical properties with the nearby Monoceros Ring. In particular, the proper motions of the EBS, like Monoceros, are indicative of prograde rotation (Vϕ ∼ 180-220 km s-1), which is similar to the Galactic thick disc. The kinematic structure of stars in the vicinity of the EBS suggests that it is not a distinct stellar stream, but rather marks the 'edge' of the Monoceros Ring. The EBS and Hydra I are the latest substructures to be linked with Monoceros, leaving the Galactic anti-centre a mess of interlinked overdensities which likely share a unified, Galactic disc origin.

The Proper Motion of Palomar 5

NASA Astrophysics Data System (ADS)

Fritz, T. K.; Kallivayalil, N.

2015-10-01

Palomar 5 (Pal 5) is a faint halo globular cluster associated with narrow tidal tails. It is a useful system to understand the process of tidal dissolution, as well as to constrain the potential of the Milky Way. A well-determined orbit for Pal 5 would enable detailed study of these open questions. We present here the first CCD-based proper motion measurement of Pal 5 obtained using SDSS as a first epoch and new Large Binocular Telescope/Large Binocular Camera (LBC) images as a second, giving a baseline of 15 years. We perform relative astrometry, using SDSS as a distortion-free reference, and images of the cluster and also of the Pal 5 stream for the derivation of the distortion correction for LBC. The reference frame is made up of background galaxies. We correct for differential chromatic refraction using relations obtained from SDSS colors as well as from flux-calibrated spectra, finding that the correction relations for stars and for galaxies are different. We obtain μα = -2.296 ± 0.186 mas yr-1 and μδ = -2.257 ± 0.181 mas yr-1 for the proper motion of Pal 5. We use this motion, and the publicly available code galpy, to model the disruption of Pal 5 in different Milky Way models consisting of a bulge, a disk, and a spherical dark matter halo. Our fits to the observed stream properties (streak and radial velocity gradient) result in a preference for a relatively large Pal 5 distance of around 24 kpc. A slightly larger absolute proper motion than what we measure also results in better matches but the best solutions need a change in distance. We find that a spherical Milky Way model, with V0 = 220 km s-1 and V20 kpc, i.e., approximately at the apocenter of Pal 5, of 218 km s-1, can match the data well, at least for our choice of disk and bulge parametrization. Based on LBT data. The LBT is an international collaboration among institutions in the United States, Italy and Germany. LBT Corporation partners are: The Ohio State University, and The Research

Improvement of Baltic proper water quality using large-scale ecological engineering.

PubMed

Stigebrandt, Anders; Gustafsson, Bo G

2007-04-01

Eutrophication of the Baltic proper has led to impaired water quality, demonstrated by, e.g., extensive blooming of cyanobacteria during the premium summer holiday season and severe oxygen deficit in the deepwater. Sustainable improvements in water quality by the reduction of phosphorus (P) supplies will take several decades before giving full effects because of large P storages both in soils in the watershed and in the water column and bottom sediments of the Baltic proper. In this article it is shown that drastically improved water quality may be obtained within a few years using large-scale ecological engineering methods. Natural variations in the Baltic proper during the last decades have demonstrated how rapid improvements may be achieved. The present article describes the basic dynamics of P, organic matter, and oxygen in the Baltic proper. It also briefly discusses the advantages and disadvantages of different classes of methods of ecological engineering aimed at restoring the Baltic proper from eutrophication effects. Preliminary computations show that the P content might be halved within a few years if about 100 kg O2 s(-1) are supplied to the upper deepwater. This would require 100 pump stations, each transporting about 100 m3 s(-1) of oxygen-rich so-called winter water from about 50 to 125 m depth where the water is released as a buoyant jet. Each pump station needs a power supply of 0.6 MW. Offshore wind power technology seems mature enough to provide the power needed by the pump stations. The cost to install 100 wind-powered pump stations, each with 0.6 MW power, at about 125-m depth is about 200 million Euros.

Using learning theory, interprofessional facilitation competencies, and behavioral indicators to evaluate facilitator training.

PubMed

LeGros, Theresa A; Amerongen, Helen M; Cooley, Janet H; Schloss, Ernest P

2015-01-01

Despite the increasing need for faculty and preceptors skilled in interprofessional facilitation (IPF), the relative novelty of the field poses a challenge to the development and evaluation of IPF programs. We use learning theory and IPF competencies with associated behavioral indicators to develop and evaluate six key messages in IPF training and experience. Our mixed methods approach included two phases: quantitative data collection with embedded qualitative data, followed by qualitative data collection in explanatory sequential fashion. This enabled triangulated analyses of both data types and of facilitation behaviors from facilitator and student perspectives. Results indicate the competency-based training was effective. Facilitators felt comfortable performing behaviors associated with IPF competencies; student observations of those behaviors supported facilitator self-reported performance. Overall, students perceived more facilitation opportunities than facilitators. Findings corroborate the importance of recruiting seasoned facilitators and establishing IPF guidelines that acknowledge variable team dynamics and help facilitators recognize teachable moments.

49 CFR 109.11 - Assistance of properly qualified personnel.

Code of Federal Regulations, 2014 CFR

2014-10-01

... MATERIALS SAFETY ADMINISTRATION, DEPARTMENT OF TRANSPORTATION HAZARDOUS MATERIALS AND OIL TRANSPORTATION... conducted under this part if the agent is not properly qualified to perform a function that is essential to...

49 CFR 109.11 - Assistance of properly qualified personnel.

Code of Federal Regulations, 2011 CFR

2011-10-01

... MATERIALS SAFETY ADMINISTRATION, DEPARTMENT OF TRANSPORTATION HAZARDOUS MATERIALS AND OIL TRANSPORTATION... this part if the agent is not properly qualified to perform a function that is essential to the agent's...

49 CFR 109.11 - Assistance of properly qualified personnel.

Code of Federal Regulations, 2012 CFR

2012-10-01

... MATERIALS SAFETY ADMINISTRATION, DEPARTMENT OF TRANSPORTATION HAZARDOUS MATERIALS AND OIL TRANSPORTATION... this part if the agent is not properly qualified to perform a function that is essential to the agent's...

Molecular mechanisms of homologous chromosome pairing and segregation in plants.

PubMed

Zhang, Jing; Zhang, Bing; Su, Handong; Birchler, James A; Han, Fangpu

2014-03-20

In most eukaryotic species, three basic steps of pairing, recombination and synapsis occur during prophase of meiosis I. Homologous chromosomal pairing and recombination are essential for accurate segregation of chromosomes. In contrast to the well-studied processes such as recombination and synapsis, many aspects of chromosome pairing are still obscure. Recent progress in several species indicates that the telomere bouquet formation can facilitate homologous chromosome pairing by bringing chromosome ends into close proximity, but the sole presence of telomere clustering is not sufficient for recognizing homologous pairs. On the other hand, accurate segregation of the genetic material from parent to offspring during meiosis is dependent on the segregation of homologs in the reductional meiotic division (MI) with sister kinetochores exhibiting mono-orientation from the same pole, and the segregation of sister chromatids during the equational meiotic division (MII) with kinetochores showing bi-orientation from the two poles. The underlying mechanism of orientation and segregation is still unclear. Here we focus on recent studies in plants and other species that provide insight into how chromosomes find their partners and mechanisms mediating chromosomal segregation. Copyright © 2013. Published by Elsevier Ltd.

32 CFR 842.62 - Who are proper claimants.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 32 National Defense 6 2014-07-01 2014-07-01 false Who are proper claimants. 842.62 Section 842.62 National Defense Department of Defense (Continued) DEPARTMENT OF THE AIR FORCE CLAIMS AND LITIGATION..., including a municipal and prefectural government. (e) Foreign companies and business entities. ...

32 CFR 842.62 - Who are proper claimants.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 32 National Defense 6 2013-07-01 2013-07-01 false Who are proper claimants. 842.62 Section 842.62 National Defense Department of Defense (Continued) DEPARTMENT OF THE AIR FORCE CLAIMS AND LITIGATION..., including a municipal and prefectural government. (e) Foreign companies and business entities. ...

32 CFR 842.62 - Who are proper claimants.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 32 National Defense 6 2012-07-01 2012-07-01 false Who are proper claimants. 842.62 Section 842.62 National Defense Department of Defense (Continued) DEPARTMENT OF THE AIR FORCE CLAIMS AND LITIGATION..., including a municipal and prefectural government. (e) Foreign companies and business entities. ...

32 CFR 842.62 - Who are proper claimants.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 32 National Defense 6 2011-07-01 2011-07-01 false Who are proper claimants. 842.62 Section 842.62 National Defense Department of Defense (Continued) DEPARTMENT OF THE AIR FORCE CLAIMS AND LITIGATION..., including a municipal and prefectural government. (e) Foreign companies and business entities. ...

Positions and proper motions of dwarf carbon stars

NASA Technical Reports Server (NTRS)

Deutsch, Eric W.

1994-01-01

Recent-epochs positions and proper motions of nine dwarf carbon star candidates are presented along with finding charts for each object. Measurements are obtained from digitized Palomar Observatory Sky Survey (POSS) and Quik V plate archives at the Space Telescope Science Institute, and from recent CCD images.

SPERMATID MICRONUCLEUS ANALYSES OF TRICHLOROETHYLENE AND CHLORAL HYDRATE EFFECTS IN MICE

EPA Science Inventory

Mice were exposed by inhalation to trichloroethylene (TCE), or by i.p. injection to the TCE metabolite, chloral hydrate (CH). arly spermatids were analyzed for micronucleus (MN) frequency and kinetochore status (presence or absence) using fluorochrome-labeled anti-kinetochore ant...

Iranian Nurses' Views on Barriers and Facilitators in Patient Education: A Cross-Sectional Study.

PubMed

Ramezanli, Somayeh; Badiyepeymaie Jahromi, Zohreh

2015-03-18

As a major factor in patient-centered care, patient education has a great impact on the quality of care provided by nurses; however, clinical nurses' performance with regard to patient education is not satisfactory. This study is an attempt to investigate barriers and facilitators in patient education from nurses' point of view. 122 nurses at Jahrom University of Medical Sciences participated in this descriptive-cross sectional study. Sampling was based on the census method. The questionnaire used to collect data included questions about nurses' demography, barriers (10 questions), and facilitators (10 questions) in patient education. The questionnaire was designed to be completed independently. To analyze the data, the researchers used descriptive statistics, including frequency, mean and standard deviation. The highest scores related to barriers to patient education were: nurses' insufficient knowledge, patients' physical and emotional unpreparedness, and lack of a proper environment for education. The most important facilitators, on the other hand, were: enhancement of instructing nurses' knowledge and skills, motivating nurses, and a step-by-step approach to patient education. It is important that nurses be prepared and motivated to train their patients. By satisfactory patient education on the part of nurses, patients will be more willing to cooperate in the treatment process.

Iranian Nurses’ Views on Barriers and Facilitators in Patient Education: A Cross-Sectional Study

PubMed Central

Ramezanli, Somayeh; Jahromi, Zohreh Badiyepeymaie

2015-01-01

Background: As a major factor in patient-centered care, patient education has a great impact on the quality of care provided by nurses; however, clinical nurses’ performance with regard to patient education is not satisfactory. This study is an attempt to investigate barriers and facilitators in patient education from nurses’ point of view. Methods: 122 nurses at Jahrom University of Medical Sciences participated in this descriptive-cross sectional study. Sampling was based on the census method. The questionnaire used to collect data included questions about nurses’ demography, barriers (10 questions), and facilitators (10 questions) in patient education. The questionnaire was designed to be completed independently. To analyze the data, the researchers used descriptive statistics, including frequency, mean and standard deviation. Results: The highest scores related to barriers to patient education were: nurses’ insufficient knowledge, patients’ physical and emotional unpreparedness, and lack of a proper environment for education. The most important facilitators, on the other hand, were: enhancement of instructing nurses’ knowledge and skills, motivating nurses, and a step-by-step approach to patient education. Conclusion: It is important that nurses be prepared and motivated to train their patients. By satisfactory patient education on the part of nurses, patients will be more willing to cooperate in the treatment process. PMID:26156926

Proper Use of Audio-Visual Aids: Essential for Educators.

ERIC Educational Resources Information Center

Dejardin, Conrad

1989-01-01

Criticizes educators as the worst users of audio-visual aids and among the worst public speakers. Offers guidelines for the proper use of an overhead projector and the development of transparencies. (DMM)

In use performance of catalytic converters on properly maintained high mileage vehicles

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sabourin, M.A.; Larson, R.E.; Donahue, K.S.

1986-01-01

A test program to evaluate the performance of catalytic converters from fifty-six 1981 and 1982 model year high mileage properly maintained in-use vehicles (from 21 engine families) was performed by the Certification Division of the Office of Mobile Sources (EPA). The program is called the Catalyst Change Program. All program vehicles were screened for proper maintenance and for mileages that ranged from 35,000 to 60,000 miles. Among vehicles belonging to 21 high sales volume and high technology engine and emission control system designs tested, poor catalyst performance was determined to be a significant contributor to emissions failure of properly-maintained vehiclesmore » at or near their warranted useful life mileage.« less

RELATIVE PROPER MOTIONS IN THE RHO OPHIUCHI CLUSTER

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wilking, Bruce A.; Sullivan, Timothy; Vrba, Frederick J., E-mail: bwilking@umsl.edu, E-mail: tsullivan@umsl.edu, E-mail: fjv@nofs.navy.mil

2015-12-10

Near-infrared images optimized for astrometry have been obtained for four fields in the high-density L 1688 cloud core over a 12 year period. The targeted regions include deeply embedded young stellar objects (YSOs) and very low luminosity objects too faint and/or heavily veiled for spectroscopy. Relative proper motions in R.A. and decl. were computed for 111 sources and again for a subset of 65 YSOs, resulting in a mean proper motion of (0,0) for each field. Assuming each field has the same mean proper motion, YSOs in the four fields were combined to yield estimates of the velocity dispersions inmore » R.A. and decl. that are consistent with 1.0 km s{sup −1}. These values appear to be independent of the evolutionary state of the YSOs. The observed velocity dispersions are consistent with the dispersion in radial velocity derived for optically visible YSOs at the periphery of the cloud core and are consistent with virial equilibrium. The higher velocity dispersion of the YSOs in the plane of the sky relative to that of dense cores may be a consequence of stellar encounters due to dense cores and filaments fragmenting to form small groups of stars or the global collapse of the L 1688 cloud core. An analysis of the differential magnitudes of objects over the 12 year baseline has not only confirmed the near-infrared variability for 29 YSOs established by prior studies, but has also identified 18 new variability candidates. Four of these have not been previously identified as YSOs and may be newly identified cluster members.« less

A Documentation System to Save Time and Ensure Proper Application of the Fiberoptic Endoscopic Evaluation of Swallowing (FEES®)

PubMed Central

Hey, Christiane; Pluschinski, Petra; Stanschus, Soenke; Euler, Harald A.; Sader, Robert A.; Langmore, Susan; Neumann, Katrin

2011-01-01

A properly performed fiberoptic endoscopic evaluation of swallowing (FEES®) is comprehensive and time-consuming. Editing times of FEES protocols and attempts for efficiency maximization are unknown. Here, the protocol editing times of completed FEES examinations were determined. The present study reports the time savings and quality gains of a newly developed documentation system tailored to the FEES standard of Langmore. Four independent examiners analyzed twelve videos of FEES procedures, six without and six with the documentation system. Effectiveness of the documentation system was evaluated according to the times for total evaluation, interpretation, documentation, report writing, and for report completeness. The documentation system reduced editing times and increased report completeness with large effect sizes. Averaged total evaluation time decreased from 42 to 27 min, report completeness increased from 55 to 80%. The use of the documentation system facilitates and improves the assessment of the swallowing process. PMID:20938202

32 CFR 842.62 - Who are proper claimants.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 32 National Defense 6 2010-07-01 2010-07-01 false Who are proper claimants. 842.62 Section 842.62 National Defense Department of Defense (Continued) DEPARTMENT OF THE AIR FORCE CLAIMS AND LITIGATION... inhabitants of a foreign country who are: (a) Foreign nationals. (b) US nationals, unless they reside there...

Proper Installation of New Windows | Efficient Windows Collaborative

Science.gov Websites

airtight fit and avoid water leakage. Always follow manufacturers installation guidelines and use trained professionals for window installation. Proper installation will: Protect from water damage. Windows should form a continuous water barrier where they meet the wall. With improper installation, water may penetrate

RCC1 regulates inner centromeric composition in a Ran-independent fashion.

PubMed

Zhang, Michael Shaofei; Furuta, Maiko; Arnaoutov, Alexei; Dasso, Mary

2018-01-01

RCC1 associates to chromatin dynamically within mitosis and catalyzes Ran-GTP production. Exogenous RCC1 disrupts kinetochore structure in Xenopus egg extracts (XEEs), but the molecular basis of this disruption remains unknown. We have investigated this question, utilizing replicated chromosomes that possess paired sister kinetochores. We find that exogenous RCC1 evicts a specific subset of inner KT proteins including Shugoshin-1 (Sgo1) and the chromosome passenger complex (CPC). We generated RCC1 mutants that separate its enzymatic activity and chromatin binding. Strikingly, Sgo1 and CPC eviction depended only on RCC1's chromatin affinity but not its capacity to produce Ran-GTP. RCC1 similarly released Sgo1 and CPC from synthetic kinetochores assembled on CENP-A nucleosome arrays. Together, our findings indicate RCC1 regulates kinetochores at the metaphase-anaphase transition through Ran-GTP-independent displacement of Sgo1 and CPC.

Observations on the Proper Orthogonal Decomposition

NASA Technical Reports Server (NTRS)

Berkooz, Gal

1992-01-01

The Proper Orthogonal Decomposition (P.O.D.), also known as the Karhunen-Loeve expansion, is a procedure for decomposing a stochastic field in an L(2) optimal sense. It is used in diverse disciplines from image processing to turbulence. Recently the P.O.D. is receiving much attention as a tool for studying dynamics of systems in infinite dimensional space. This paper reviews the mathematical fundamentals of this theory. Also included are results on the span of the eigenfunction basis, a geometric corollary due to Chebyshev's inequality and a relation between the P.O.D. symmetry and ergodicity.

Chromosome congression is promoted by CENP-Q- and CENP-E-dependent pathways

PubMed Central

Bancroft, James; Auckland, Philip; Samora, Catarina P.; McAinsh, Andrew D.

2015-01-01

ABSTRACT A key step of mitosis is the congression of chromosomes to the spindle equator. Congression is driven by at least two distinct mechanisms: (1) kinetochores slide along the microtubule lattice using the plus-end directed CENP-E motor, and (2) kinetochores biorientating near the pole move to the equator through microtubule depolymerisation-coupled pulling. Here, we show that CENP-Q – a subunit of the CENP-O complex (comprising CENP-O, CENP-P, CENP-Q and CENP-U) that targets polo-like kinase (Plk1) to kinetochores – is also required for the recruitment of CENP-E to kinetochores. We further reveal a CENP-E recruitment-independent role for CENP-Q in depolymerisation-coupled pulling. Both of these functions are abolished by a single point mutation in CENP-Q (S50A) – a residue that is phosphorylated in vivo. Importantly, the S50A mutant does not affect the loading of Plk1 onto kinetochores and leaves the CENP-O complex intact. Thus, the functions of CENP-Q in CENP-E loading and depolymerisation-coupled pulling are independent from its role in Plk1 recruitment and CENP-O complex stabilisation. Taken together, our data provide evidence that phosphoregulation of CENP-Q plays a central function in coordinating chromosome congression mechanisms. PMID:25395579

Mechanisms of Chromosome Congression during Mitosis

PubMed Central

Maiato, Helder; Gomes, Ana Margarida; Sousa, Filipe; Barisic, Marin

2017-01-01

Chromosome congression during prometaphase culminates with the establishment of a metaphase plate, a hallmark of mitosis in metazoans. Classical views resulting from more than 100 years of research on this topic have attempted to explain chromosome congression based on the balance between opposing pulling and/or pushing forces that reach an equilibrium near the spindle equator. However, in mammalian cells, chromosome bi-orientation and force balance at kinetochores are not required for chromosome congression, whereas the mechanisms of chromosome congression are not necessarily involved in the maintenance of chromosome alignment after congression. Thus, chromosome congression and maintenance of alignment are determined by different principles. Moreover, it is now clear that not all chromosomes use the same mechanism for congressing to the spindle equator. Those chromosomes that are favorably positioned between both poles when the nuclear envelope breaks down use the so-called “direct congression” pathway in which chromosomes align after bi-orientation and the establishment of end-on kinetochore-microtubule attachments. This favors the balanced action of kinetochore pulling forces and polar ejection forces along chromosome arms that drive chromosome oscillatory movements during and after congression. The other pathway, which we call “peripheral congression”, is independent of end-on kinetochore microtubule-attachments and relies on the dominant and coordinated action of the kinetochore motors Dynein and Centromere Protein E (CENP-E) that mediate the lateral transport of peripheral chromosomes along microtubules, first towards the poles and subsequently towards the equator. How the opposite polarities of kinetochore motors are regulated in space and time to drive congression of peripheral chromosomes only now starts to be understood. This appears to be regulated by position-dependent phosphorylation of both Dynein and CENP-E and by spindle microtubule

MiR-520d-3p antitumor activity in human breast cancer via post-transcriptional regulation of spindle and kinetochore associated 2 expression.

PubMed

Ren, Zhouhui; Yang, Tong; Ding, Jie; Liu, Weihong; Meng, Xiangyu; Zhang, Pingping; Liu, Kaitai; Wang, Ping

2018-01-01

MicroRNAs (miRNAs) play an important role in human tumorigenesis as oncogenes or tumor suppressors by directly binding to the 3'-untranslated region of their target mRNAs. MiR-520d-3p has been reported as a tumor suppressor gene in ovarian cancer and gastric cancer, while the function of miR-520d-3p in human breast cancers is still uninvolved. In this study, we initially identified that the expression of miR-520d-3p was significantly reduced in breast cancer specimens and cell lines. The restoration of miR-520d-3p expression not only reduced breast cancer cell viability by causing the accumulation of G2 phase and cell apoptosis, but also inhibited tumorigenicity in vivo . In addition, as a critical target of miR-520d-3p, the activity of spindle and kinetochore associated 2 (SKA2) was greatly inhibited by miR-520d-3p, and overexpression of miR-520d-3p decreased the expression of SKA2. SKA2 downregulation suppressed cell viability, whereas restoration of SKA2 expression significantly reversed the inhibitory effects of miR-520d-3p antitumor activity. Furthermore, SKA2 was frequently overexpressed in clinical specimens and cell lines, and the expression levels were statistically inversely correlated with miR-520d-3p expression. In conclusion, our data demonstrated that miR-520d-3p antitumor activity is achieved by targeting the SKA2 in human breast cancer cells, suggesting that miR-520d-3p may be a potential target molecule for the therapy.

The Absolute Proper Motion of NGC 6397 Revisited

NASA Astrophysics Data System (ADS)

Rees, Richard; Cudworth, Kyle

2018-01-01

We compare several determinations of the absolute proper motion of the Galactic globular cluster NGC 6397: (1) our own determination relative to field stars derived from scans of 38 photographic plates spanning 97 years in epoch; (2) using our proper motion membership to identify cluster stars in various catalogs in the literature (UCAC4, UCAC5, PPMXL, HSOY, Tycho-2, Hipparcos, TGAS); (3) published results from the Yale SPM Program (both tied to Hipparcos and relative to galaxies) and two from HST observations relative to galaxies. The various determinations are not in good agreement. Curiously, the Yale SPM relative to galaxies does not agree with the HST determinations, and the individual HST error ellipses are close to each other but do not overlap. The Yale SPM relative to galaxies does agree with our determination, Tycho-2, and the Yale SPM tied to Hipparcos. It is not clear which of the current determinations is most reliable; we have found evidence of systematic errors in some of them (including one of the HST determinations). This research has been partially supported by the NSF.

Organism and artifact: Proper functions in Paley organisms.

PubMed

Holm, Sune

2013-12-01

In this paper I assess the explanatory powers of theories of function in the context of products that may result from synthetic biology. The aim is not to develop a new theory of functions, but to assess existing theories of function in relation to a new kind of biological and artifactual entity that might be produced in the not-too-distant future by means of synthetic biology. The paper thus investigates how to conceive of the functional nature of living systems that are not the result of evolution by natural selection, or instantly generated by cosmic coincidence, but which are products of intelligent design. The paper argues that the aetiological theory of proper functions in organisms and artifacts is inadequate as an account of proper functions in such 'Paley organisms' and defends an alternative organisational approach. The paper ends by considering the implications of the discussion of biological function for questions about the interests and moral status of non-sentient organisms. Copyright © 2013 Elsevier Ltd. All rights reserved.

Partner-facilitating transmembrane penetration of nanoparticles: a biological test in silico.

PubMed

Wang, W; Yang, R; Zhang, F; Yuan, B; Yang, K; Ma, Y

2018-06-21

Transmembrane penetration of nanoparticles (NPs) promises an effective pathway for cargo delivery into cells, and offers the possibility of organelle-specific targeting for biomedical applications. However, a full understanding of the underlying NP-membrane interaction mechanism is still lacking. In this work, the membrane penetration behavior of NPs is statistically analyzed based on the simulations of over 2.2 ms, which are performed with dissipative particle dynamics (DPD). Influences from multiple factors including the NP concentration, shape and surface chemistry are taken into account. It is interesting to find that, the introduction of a partner NP would greatly facilitate the transmembrane penetration of a host spherical NP. This is probably due to the membrane-mediated cooperation between the NPs. Moreover, the proper selection of a partner NP with specific surface chemistry is of great significance. For example, the best partner for a hydrophilic NP to achieve transmembrane penetration is a Janus-like one, in comparison with the hydrophilic, hydrophobic or randomly surface-decorated NPs. Furthermore, such a partner-facilitating effect in NP translocation also works for a shaped NP although less pronounced. Our results are helpful for a better understanding of the complicated nano-bio interactions, and offer a practical guide to the NP-based drug delivery strategy with high efficiency.

Treatment of Proper Name Retrieval Deficits in an Individual with Temporal Lobe Epilepsy

ERIC Educational Resources Information Center

Minkina, Irene; Ojemann, Jeffrey G.; Grabowski, Thomas J.; Silkes, JoAnn P.; Phatak, Vaishali; Kendall, Diane L.

2013-01-01

Purpose: Studies investigating language deficits in individuals with left temporal-lobe epilepsy have consistently demonstrated impairments in proper name retrieval. The aim of this Phase I rehabilitation study was to investigate the effects of a linguistically distributed word retrieval treatment on proper name retrieval in an individual with…

Stable kinetochore–microtubule attachment is sufficient to silence the spindle assembly checkpoint in human cells

PubMed Central

Tauchman, Eric C.; Boehm, Frederick J.; DeLuca, Jennifer G.

2015-01-01

During mitosis, duplicated sister chromatids attach to microtubules emanating from opposing sides of the bipolar spindle through large protein complexes called kinetochores. In the absence of stable kinetochore–microtubule attachments, a cell surveillance mechanism known as the spindle assembly checkpoint (SAC) produces an inhibitory signal that prevents anaphase onset. Precisely how the inhibitory SAC signal is extinguished in response to microtubule attachment remains unresolved. To address this, we induced formation of hyper-stable kinetochore–microtubule attachments in human cells using a non-phosphorylatable version of the protein Hec1, a core component of the attachment machinery. We find that stable attachments are sufficient to silence the SAC in the absence of sister kinetochore bi-orientation and strikingly in the absence of detectable microtubule pulling forces or tension. Furthermore, we find that SAC satisfaction occurs despite the absence of large changes in intra-kinetochore distance, suggesting that substantial kinetochore stretching is not required for quenching the SAC signal. PMID:26620470

Update of membership and mean proper motion of open clusters from UCAC5 catalog

NASA Astrophysics Data System (ADS)

Dias, W. S.; Monteiro, H.; Assafin, M.

2018-06-01

We present mean proper motions and membership probabilities of individual stars for optically visible open clusters, which have been determined using data from the UCAC5 catalog. This follows our previous studies with the UCAC2 and UCAC4 catalogs, but now using improved proper motions in the GAIA reference frame. In the present study results were obtained for a sample of 1108 open clusters. For five clusters, this is the first determination of mean proper motion, and for the whole sample, we present results with a much larger number of identified astrometric member stars than on previous studies. It is the last update of our Open cluster Catalog based on proper motion data only. Future updates will count on astrometric, photometric and spectroscopic GAIA data as input for analyses.

A 1500 deg2 near infrared proper motion catalogue from the UKIDSS Large Area Survey

NASA Astrophysics Data System (ADS)

Smith, Leigh; Lucas, P. W.; Burningham, B.; Jones, H. R. A.; Smart, R. L.; Andrei, A. H.; Catalán, S.; Pinfield, D. J.

2014-02-01

The United Kingdom Infrared Deep Sky Survey (UKIDSS) Large Area Survey (LAS) began in 2005, with the start of the UKIDSS programme as a 7 year effort to survey roughly 4000 deg2 at high Galactic latitudes in Y, J, H and K bands. The survey also included a significant quantity of two epoch J band observations, with an epoch baseline greater than 2 years to calculate proper motions. We present a near-infrared proper motion catalogue for the 1500 deg2 of the two epoch LAS data, which includes 135 625 stellar sources and a further 88 324 with ambiguous morphological classifications, all with motions detected above the 5σ level. We developed a custom proper motion pipeline which we describe here. Our catalogue agrees well with the proper motion data supplied for a 300 deg2 subset in the current Wide Field Camera Science Archive (WSA) 10th data release (DR10) catalogue, and in various optical catalogues, but it benefits from a larger matching radius and hence a larger upper proper motion detection limit. We provide absolute proper motions, using LAS galaxies for the relative to absolute correction. By using local second-order polynomial transformations, as opposed to linear transformations in the WSA, we correct better for any local distortions in the focal plane, not including the radial distortion that is removed by the UKIDSS pipeline. We present the results of proper motion searches for new brown dwarfs and white dwarfs. We discuss 41 sources in the WSA DR10 overlap with our catalogue with proper motions >300 mas yr-1, several of which are new detections. We present 15 new candidate ultracool dwarf binary systems.

Understanding Facilitation: Theory and Principles.

ERIC Educational Resources Information Center

Hogan, Christine

This book introduces newcomers to the concept of facilitation, and it presents a critical analysis of established and current theory on facilitation for existing practitioners. The following are among the topics discussed: (1) emergence of the field of facilitation; (2) development of facilitation in management; (3) development of facilitation in…

Mitotic Kinesin CENP-E Promotes Microtubule Plus-End Elongation

PubMed Central

Sardar, Harjinder S.; Luczak, Vincent G.; Lopez, Maria M.; Lister, Bradford C.; Gilbert, Susan P.

2010-01-01

Summary Centromere protein CENP-E is a dimeric kinesin (Kinesin-7 family) with critical roles in mitosis including establishment of microtubule (MT)-chromosome linkage and movement of monooriented chromosomes on kinetochore microtubules (kMTs) for proper alignment at metaphase [1-9]. We performed studies to test the hypothesis that CENP-E promotes MT elongation at the MT plus-ends. A human CENP-E construct was engineered, expressed, and purified which yielded the CENP-E-6His dimeric motor protein. The results show that CENP-E promotes MT plus-end directed MT gliding at 11 nm/s. The results from real-time microscopy assays indicate that 60.3% of polarity marked MTs exhibited CENP-E promoted MT plus-end elongation. The MT extension required ATP turnover, and MT plus-end elongation occurred at 1.48 μm/30 min. Immunolocalization studies revealed that 80.8% of plus-end elongated MTs showed CENP-E at the MT plus-end. The time dependence of CENP-E promoted MT elongation in solution best fit a single exponential function (kobs = 5.1 s−1), which is indicative of a mechanism in which α,β-tubulin subunit addition is tightly coupled to ATP turnover. Based on these results, we propose that CENP-E as part of its function in chromosome kinetochore-MT linkage plays a direct role in MT elongation. PMID:20797864

P38 Mitogen-activated Protein Kinase Activity Is Required during Mitosis for Timely Satisfaction of the Mitotic Checkpoint But Not for the Fidelity of Chromosome Segregation

PubMed Central

Lee, Kyunghee; Kenny, Alison E.

2010-01-01

Although p38 activity is reported to be required as cells enter mitosis for proper spindle assembly and checkpoint function, its role during the division process remains controversial in lieu of direct data. We therefore conducted live cell studies to determine the effect on mitosis of inhibiting or depleting p38. We found that in the absence of p38 activity the duration of mitosis is prolonged by ∼40% in nontransformed human RPE-1, ∼80% in PtK2 (rat kangaroo), and ∼25% in mouse cells, and this prolongation leads to an elevated mitotic index. However, under this condition chromatid segregation and cytokinesis are normal. Using Mad2/YFP-expressing cells, we show the prolongation of mitosis in the absence of p38 activity is directly due to a delay in satisfying the mitotic checkpoint. Inhibiting p38 did not affect the rate of chromosome motion; however, it did lead to the formation of significantly (10%) longer metaphase spindles. From these data we conclude that normal p38 activity is required for the timely stable attachment of all kinetochores to spindle microtubules, but not for the fidelity of the mitotic process. We speculate that p38 activity promotes timely checkpoint satisfaction by indirectly influencing those motor proteins (e.g., Klp10, Klp67A) involved in regulating the dynamics of kinetochore microtubule ends. PMID:20462950

Optical identification of two nearby Isolated Neutron Stars through proper motion measuremnt.

NASA Astrophysics Data System (ADS)

Zane, Silvia

2004-07-01

Aim of this proposal is to perform high-resolution imaging of the proposed optical counterparts of the two, radio silent, isolated neutron stars RXJ1308.6+2127 and RX J1605.3+3249 with the STIS/50CCD. Imaging both fields with the same instrumental configuration used in mid 2001 by Kaplan et al {2002; 2003}, will allow us to measure the objects' position and to determine their proper motions over a time base of nearly four years. The measurement of proper motions at the level of at least few tens mas/yr, expected for relatively nearby neutron stars, would unambigouosly secure the proposed optical identifications, not achievable otherwise. In addition, the knowledge of the proper motion will provide useful indications on the space velocity and distance of these neutrons stars, as well as on the radius. Constraining these parameters is of paramount importance to discriminate between the variety of emission mechanisms invoked to explain their observed thermal X-ray spectra and to probe the neutron star equation of state {EOS}. The determination of the proper motion is a decisive step toward a dedicated follow-up program aimed at measuring the objects' optical parallax, thus providing much firmer constrains on the star properties, again to be performed with the STIS/50CCD.

Proper motion and secular variations of Keplerian orbital elements

NASA Astrophysics Data System (ADS)

Butkevich, Alexey G.

2018-05-01

High-precision observations require accurate modelling of secular changes in the orbital elements in order to extrapolate measurements over long time intervals, and to detect deviation from pure Keplerian motion caused, for example, by other bodies or relativistic effects. We consider the evolution of the Keplerian elements resulting from the gradual change of the apparent orbit orientation due to proper motion. We present rigorous formulae for the transformation of the orbit inclination, longitude of the ascending node and argument of the pericenter from one epoch to another, assuming uniform stellar motion and taking radial velocity into account. An approximate treatment, accurate to the second-order terms in time, is also given. The proper motion effects may be significant for long-period transiting planets. These theoretical results are applicable to the modelling of planetary transits and precise Doppler measurements as well as analysis of pulsar and eclipsing binary timing observations.

Proper Motion of Components in 4C 39.25

NASA Technical Reports Server (NTRS)

Guirado, J. C.; Marcaide, J. M.; Alberdi, A.; Elosegui, P.; Ratner, M. I.; Shapiro, I. I.; Kilger, R.; Mantovani, F.; Venturi, T.; Rius, A.;

A Perikinetochoric Ring Defined by MCAK and Aurora-B as a Novel Centromere Domain

PubMed Central

Parra, María Teresa; Gómez, Rocío; Viera, Alberto; Page, Jesús; Calvente, Adela; Wordeman, Linda; Rufas, Julio S; Suja, José A

2006-01-01

Mitotic Centromere-Associated Kinesin (MCAK) is a member of the kinesin-13 subfamily of kinesin-related proteins. In mitosis, this microtubule-depolymerising kinesin seems to be implicated in chromosome segregation and in the correction of improper kinetochore-microtubule interactions, and its activity is regulated by the Aurora-B kinase. However, there are no published data on its behaviour and function during mammalian meiosis. We have analysed by immunofluorescence in squashed mouse spermatocytes, the distribution and possible function of MCAK, together with Aurora-B, during both meiotic divisions. Our results demonstrate that MCAK and Aurora-B colocalise at the inner domain of metaphase I centromeres. Thus, MCAK shows a “cone”-like three-dimensional distribution beneath and surrounding the closely associated sister kinetochores. During the second meiotic division, MCAK and Aurora-B also colocalise at the inner centromere domain as a band that joins sister kinetochores, but only during prometaphase II in unattached chromosomes. During chromosome congression to the metaphase II plate, MCAK relocalises and appears as a ring below each sister kinetochore. Aurora-B also relocalises to appear as a ring surrounding and beneath kinetochores but during late metaphase II. Our results demonstrate that the redistribution of MCAK at prometaphase II/metaphase II centromeres depends on tension across the centromere and/or on the interaction of microtubules with kinetochores. We propose that the perikinetochoric rings of MCAK and Aurora-B define a novel transient centromere domain at least in mouse chromosomes during meiosis. We discuss the possible functions of MCAK at the inner centromere domain and at the perikinetochoric ring during both meiotic divisions. PMID:16741559

CENPT bridges adjacent CENPA nucleosomes on young human α-satellite dimers

PubMed Central

Thakur, Jitendra; Henikoff, Steven

2016-01-01

Nucleosomes containing the CenH3 (CENPA or CENP-A) histone variant replace H3 nucleosomes at centromeres to provide a foundation for kinetochore assembly. CENPA nucleosomes are part of the constitutive centromere associated network (CCAN) that forms the inner kinetochore on which outer kinetochore proteins assemble. Two components of the CCAN, CENPC and the histone-fold protein CENPT, provide independent connections from the ∼171-bp centromeric α-satellite repeat units to the outer kinetochore. However, the spatial relationship between CENPA nucleosomes and these two branches remains unclear. To address this issue, we use a base-pair resolution genomic readout of protein–protein interactions, comparative chromatin immunoprecipitation (ChIP) with sequencing, together with sequential ChIP, to infer the in vivo molecular architecture of the human CCAN. In contrast to the currently accepted model in which CENPT associates with H3 nucleosomes, we find that CENPT is centered over the CENPB box between two well-positioned CENPA nucleosomes on the most abundant centromeric young α-satellite dimers and interacts with the CENPB/CENPC complex. Upon cross-linking, the entire CENPA/CENPB/CENPC/CENPT complex is nuclease-protected over an α-satellite dimer that comprises the fundamental unit of centromeric chromatin. We conclude that CENPA/CENPC and CENPT pathways for kinetochore assembly are physically integrated over young α-satellite dimers. PMID:27384170

A 2 epoch proper motion catalogue from the UKIDSS Large Area Survey

NASA Astrophysics Data System (ADS)

Smith, Leigh; Lucas, Phil; Burningham, Ben; Jones, Hugh; Pinfield, David; Smart, Ricky; Andrei, Alexandre

2013-04-01

The UKIDSS Large Area Survey (LAS) began in 2005, with the start of the UKIDSS program as a 7 year effort to survey roughly 4000 square degrees at high galactic latitudes in Y, J, H and K bands. The survey also included a significant quantity of 2-epoch J band observations, with epoch baselines ranging from 2 to 7 years. We present a proper motion catalogue for the 1500 square degrees of the 2 epoch LAS data, which includes some 800,000 sources with motions detected above the 5σ level. We developed a bespoke proper motion pipeline which applies a source-unique second order polynomial transformation to UKIDSS array coordinates of each source to counter potential local non-uniformity in the focal plane. Our catalogue agrees well with the proper motion data supplied in the current WFCAM Science Archive (WSA) DR9 catalogue where there is overlap, and in various optical catalogues, but it benefits from some improvements. One improvement is that we provide absolute proper motions, using LAS galaxies for the relative to absolute correction. Also, by using unique, local, 2nd order polynomial tranformations, as opposed to the linear transformations in the WSA, we correct better for any local distortions in the focal plane, not including the radial distortion that is removed by their pipeline.

A properly adjusted forage harvester can save time and money

USDA-ARS?s Scientific Manuscript database

A properly adjusted forage harvester can save fuel and increase the realizable milk per ton of your silage. This article details the adjustments necessary to minimize energy while maximizing productivity and forage quality....

Facilitated versus Non-Facilitated Online Case Discussions: Comparing Differences in Problem Space Coverage

ERIC Educational Resources Information Center

Ertmer, Peggy A.; Koehler, Adrie A.

2015-01-01

The facilitator plays a key role in guiding students' efforts during case discussions. However, few studies have compared differences in learning outcomes for students participating in facilitated versus non-facilitated discussions. In this research, we used "problem space coverage" as a learning measure to compare outcomes between…

32 CFR 842.63 - Who are not proper claimants.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 32 National Defense 6 2010-07-01 2010-07-01 false Who are not proper claimants. 842.63 Section 842.63 National Defense Department of Defense (Continued) DEPARTMENT OF THE AIR FORCE CLAIMS AND... accompanying US military and US national civilian employees. (c) Foreign military personnel suffering property...

14 CFR 389.22 - Failure to make proper payment.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 14 Aeronautics and Space 4 2010-01-01 2010-01-01 false Failure to make proper payment. 389.22 Section 389.22 Aeronautics and Space OFFICE OF THE SECRETARY, DEPARTMENT OF TRANSPORTATION (AVIATION PROCEEDINGS) ORGANIZATION FEES AND CHARGES FOR SPECIAL SERVICES Filing and Processing License Fees § 389.22...

14 CFR 389.22 - Failure to make proper payment.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 14 Aeronautics and Space 4 2011-01-01 2011-01-01 false Failure to make proper payment. 389.22 Section 389.22 Aeronautics and Space OFFICE OF THE SECRETARY, DEPARTMENT OF TRANSPORTATION (AVIATION PROCEEDINGS) ORGANIZATION FEES AND CHARGES FOR SPECIAL SERVICES Filing and Processing License Fees § 389.22...

Demographic factors and retrieval of object and proper names after age 70

PubMed Central

Fridkin, Shimon; Ayalon, Liat

2018-01-01

Purpose This research aimed to investigate whether demographic factors are similarly related to retrieval of object and proper names. Methods The sample included 5,907 individuals above age 70 who participated in the Health and Retirement Study between 2004 and 2012. Participants were asked to name two objects as well as the US President and Vice President. Latent growth curve models examined the associations of age, education, and self-rated health with baseline levels and change trajectories in retrieval. Results Age and education were more strongly related to retrieval of proper names than to retrieval of object names, both for baseline scores and for change trajectory. Similar effects of self-rated health emerged for both types of stimuli. Conclusions The results show that examining object names and proper names together as indication of cognitive status in the HRS might overlook important differences between the two types of stimuli, in both baseline performance and longitudinal change. PMID:29370264

42 CFR 405.430 - Failure to properly opt-out.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 42 Public Health 2 2010-10-01 2010-10-01 false Failure to properly opt-out. 405.430 Section 405.430 Public Health CENTERS FOR MEDICARE & MEDICAID SERVICES, DEPARTMENT OF HEALTH AND HUMAN SERVICES MEDICARE PROGRAM FEDERAL HEALTH INSURANCE FOR THE AGED AND DISABLED Private Contracts § 405.430 Failure to...

Searching for High Proper Motion Sources Towards the Galactic Center using Convolution Neural Networks

NASA Astrophysics Data System (ADS)

Giongo Fernandes, Alexandre; Benjamin, Robert A.; Babler, Brian

2018-01-01

Two sets of infrared images of the Galactic Center region (|L|< 1 degree and |B| < 0.75 degrees) taken by the Spitzer Space Telescope in IRAC 3.6 micron and 4.5 micron bands are searched for high proper motion objects (> 100 mas/year). The two image sets come from GALCEN observations in 2005 and GLIMPSE proper observations in 2015 with matched observation modes. We use three different methods to search for these objects in extremely crowded fields: (1) comparing matched point source lists, (2) crowd sourcing by several college introductory astronomy classes in the state of Wisconsin (700 volunteers), and (3) convolutional neural networks trained using objects from the previous two methods. Before our search six high proper objects were known, four of which were found by the VVV near-infrared Galactic plane survey. We compare and describe our methods for this search, and present a preliminary catalog of high proper motions objects.

Using Standardized Patients in Continuing Medical Education Courses on Proper Prescribing of Controlled Substances

ERIC Educational Resources Information Center

Swiggart, William H.; Ghulyan, Marine V.; Dewey, Charlene M.

2012-01-01

Controlled prescription drug (CPD) abuse is an increasing threat to patient safety and health care providers (HCPs) are not adequately prepared nor do they routinely employ proper screening techniques. Using standardized patients (SPs) as an instructional strategy, the trained physicians on proper prescribing practices and SBIRT (Screening, Brief…

The VLBA Extragalactic Proper Motion Catalog and a Measurement of the Secular Aberration Drift

NASA Astrophysics Data System (ADS)

Truebenbach, Alexandra E.; Darling, Jeremy

2017-11-01

We present a catalog of extragalactic proper motions created using archival VLBI data and our own VLBA astrometry. The catalog contains 713 proper motions, with average uncertainties of ˜24 μas yr-1, including 40 new or improved proper motion measurements using relative astrometry with the VLBA. The observations were conducted in the X-band and yielded positions with uncertainties of ˜70 μas. We add 10 new redshifts using spectroscopic observations taken at Apache Point Observatory and Gemini North. With the VLBA Extragalactic Proper Motion Catalog, we detect the secular aberration drift—the apparent motion of extragalactic objects caused by the solar system’s acceleration around the Galactic center—at a 6.3σ significance. We model the aberration drift as a spheroidal dipole, with the square root of the power equal to 4.89 ± 0.77 μas yr-1, an amplitude of 1.69 ± 0.27 μas yr-1, and an apex at (275\\buildrel{\\circ}\\over{.} 2+/- 10\\buildrel{\\circ}\\over{.} 0, -29\\buildrel{\\circ}\\over{.} 4+/- 8\\buildrel{\\circ}\\over{.} 8). Our dipole model detects the aberration drift at a higher significance than some previous studies, but at a lower amplitude than expected or previously measured. The full aberration drift may be partially removed by the no-net-rotation constraint used when measuring archival extragalactic radio source positions. Like the cosmic microwave background dipole, which is induced by the observer’s motion, the aberration drift signal should be subtracted from extragalactic proper motions in order to detect cosmological proper motions, including the Hubble expansion, long-period stochastic gravitational waves, and the collapse of large-scale structure.

Machine safety: proper safeguarding techniques.

PubMed

Martin, K J

1992-06-01

1. OSHA mandates certain safeguarding of machinery to prevent accidents and protect machine operators. OSHA specifies moving parts that must be guarded and sets criteria for the guards. 2. A 1989 OSHA standard for lockout/tagout requires locking the energy source during maintenance, periodically inspecting for power transmission, and training maintenance workers. 3. In an amputation emergency, first aid for cardiopulmonary resuscitation, shock, and bleeding are the first considerations. The amputated part should be wrapped in moist gauze, placed in a sealed plastic bag, and placed in a container of 50% water and 50% ice for transport. 4. The role of the occupational health nurse in machine safety is to conduct worksite analyses to identify proper safeguarding and to communicate deficiencies to appropriate personnel; to train workers in safe work practices and observe compliance in the use of machine guards; to provide care to workers injured by machines; and to reinforce safe work practices among machine operators.

Mps1 directs the assembly of Cdc20 inhibitory complexes during interphase and mitosis to control M phase timing and spindle checkpoint signaling.

PubMed

Maciejowski, John; George, Kelly A; Terret, Marie-Emilie; Zhang, Chao; Shokat, Kevan M; Jallepalli, Prasad V

2010-07-12

The spindle assembly checkpoint (SAC) in mammals uses cytosolic and kinetochore-based signaling pathways to inhibit anaphase. In this study, we use chemical genetics to show that the protein kinase Mps1 regulates both aspects of the SAC. Human MPS1-null cells were generated via gene targeting and reconstituted with either the wild-type kinase (Mps1(wt)) or a mutant version (Mps1(as)) sensitized to bulky purine analogues. Mps1 inhibition sharply accelerated anaphase onset, such that cells completed mitosis in 12 min, and prevented Cdc20's association with either Mad2 or BubR1 during interphase, i.e., before the appearance of functional kinetochores. Furthermore, intramitotic Mps1 inhibition evicted Bub1 and all other known SAC transducers from the outer kinetochore, but contrary to a recent study, did not perturb aurora B-dependent phosphorylation. We conclude that Mps1 has two complementary roles in SAC regulation: (1) initial cytoplasmic activation of Cdc20 inhibitors and (2) recruitment of factors that promote sustained anaphase inhibition and chromosome biorientation to unattached kinetochores.

RNA as a structural and regulatory component of the centromere.

PubMed

Gent, Jonathan I; Dawe, R Kelly

2012-01-01

Despite many challenges, great progress has been made in identifying kinetochore proteins and understanding their overall functions relative to spindles and centromeric DNA. In contrast, less is known about the specialized centromeric chromatin environment and how it may be involved in regulating the assembly of kinetochore proteins. Multiple independent lines of evidence have implicated transcription and the resulting RNA as an important part of this process. Here, we summarize recent literature demonstrating the roles of centromeric RNA in regulating kinetochore assembly and maintenance. We also review literature suggesting that the process of centromeric transcription may be as important as the resulting RNA and that such transcription may be involved in recruiting the centromeric histone variant CENH3.

PCR amplification of 16S rDNA from lyophilized cell cultures facilitates studies in molecular systematics

NASA Technical Reports Server (NTRS)

Wisotzkey, J. D.; Jurtshuk, P. Jr; Fox, G. E.

1990-01-01

The sequence of the major portion of a Bacillus cycloheptanicus strain SCH(T) 16S rRNA gene is reported. This sequence suggests that B. cycloheptanicus is genetically quite distinct from traditional Bacillus strains (e.g., B. subtilis) and may be properly regarded as belonging to a different genus. The sequence was determined from DNA that was produced by direct amplification of ribosomal DNA from a lyophilized cell pellet with straightforward polymerase chain reaction (PCR) procedures. By obviating the need to revive cell cultures from the lyophile pellet, this approach facilitates rapid 16S rDNA sequencing and thereby advances studies in molecular systematics.

Proper care for the dying: a critical public issue.

PubMed Central

Crispell, K R; Gomez, C F

1987-01-01

The ability of the medical profession to sustain life, or more appropriately, to prolong dying, in patients with terminal illness, creates a most complex and controversial situation for all involved: the patient, if mentally alert; the patient's family; and the medical care team including physicians, nurses and attendants. This situation is especially complex in large acute care hospitals where medical and nursing students, residents and house officers receive advanced medical training. A major problem, prolonging the dying of the terminally ill, with its medical, legal, ethical and economic complexities now confronts American society. The problem is particularly acute in teaching hospitals, in which one finds a disproportionate number of terminally ill patients. The ability to work at these questions as a community rather than as adversaries will determine much about the ability of the health care system to respect the dignity and autonomy of those who seek aid and comfort when faced with serious illness and impending death. Better communication between the physicians, health care providers, the lawyers and ethicists must be developed in order to solve these problems. Over the next ten years society and our elected representatives will be making very demanding decisions about the use of the health dollar. One possible way to prevent increasing costs is to reach significant agreement on the proper care of the dying. Proper care for the dying is being considered, discussed, and evaluated by very thoughtful people. It is not governments which should decide who is to live or who is to die. There is the serious problem of the 'slippery slope' to euthanasia by omission if cost containment becomes the major force in formulating policy on the proper care of the dying. PMID:3612698

Taxonomic and thematic organisation of proper name conceptual knowledge.

PubMed

Crutch, Sebastian J; Warrington, Elizabeth K

2011-01-01

We report the investigation of the organisation of proper names in two aphasic patients (NBC and FBI). The performance of both patients on spoken word to written word matching tasks was inconsistent, affected by presentation rate and semantic relatedness of the competing responses, all hallmarks of a refractory semantic access dysphasia. In a series of experiments we explored the semantic relatedness effects within their proper name vocabulary, including brand names and person names. First we demonstrated the interaction between very fine grain organisation and personal experience, with one patient with a special interest in the cinema demonstrating higher error rates when identifying the names of actors working in a similar film genre (e.g., action movies: Arnold Schwarzenegger, Bruce Willis, Sylvester Stallone, Mel Gibson) than those working in different genres (e.g., Arnold Schwarzenegger, Gregory Peck, Robin Williams, Gene Kelly). Second we compared directly two potential principles of semantic organisation - taxonomic and thematic. Furthermore we considered these principles of organisation in the context of the individuals' personal knowledge base. We selected topics matching the interests and experience of each patient, namely cinema and literature (NBC) and naval history (FBI). The stimulus items were arranged in taxonomic arrays (e.g., Jane Austen, Emily Bronte, Agatha Christie), thematic arrays (e.g., Jane Austen, Pride and Prejudice, Mr Darcy), and unrelated arrays (e.g., Jane Austen, Wuthering Heights, Hercule Poirot). We documented that different patterns of taxonomic and thematic organisation were constrained by whether the individual has limited knowledge, moderate knowledge or detailed knowledge of a particular vocabulary. It is suggested that moderate proper name knowledge is primarily organised by taxonomy whereas extensive experience results in a more detailed knowledge base in which theme is a powerful organising principle.

Taxonomic and Thematic Organisation of Proper Name Conceptual Knowledge

PubMed Central

Crutch, Sebastian J.; Warrington, Elizabeth K.

2011-01-01

We report the investigation of the organisation of proper names in two aphasic patients (NBC and FBI). The performance of both patients on spoken word to written word matching tasks was inconsistent, affected by presentation rate and semantic relatedness of the competing responses, all hallmarks of a refractory semantic access dysphasia. In a series of experiments we explored the semantic relatedness effects within their proper name vocabulary, including brand names and person names. First we demonstrated the interaction between very fine grain organisation and personal experience, with one patient with a special interest in the cinema demonstrating higher error rates when identifying the names of actors working in a similar film genre (e.g. action movies: Arnold Schwarzenegger, Bruce Willis, Sylvester Stallone, Mel Gibson) than those working in different genres (e.g. Arnold Schwarzenegger, Gregory Peck, Robin Williams, Gene Kelly). Second we compared directly two potential principles of semantic organisation – taxonomic and thematic. Furthermore we considered these principles of organisation in the context of the individuals' personal knowledge base. We selected topics matching the interests and experience of each patient, namely cinema and literature (NBC) and naval history (FBI). The stimulus items were arranged in taxonomic arrays (e.g. Jane Austen, Emily Bronte, Agatha Christie), thematic arrays (e.g. Jane Austen, Pride and Prejudice, Mr Darcy), and unrelated arrays (e.g. Jane Austen, Wuthering Heights, Hercule Poirot). We documented that different patterns of taxonomic and thematic organisation were constrained by whether the individual has limited knowledge, moderate knowledge or detailed knowledge of a particular vocabulary. It is suggested that moderate proper name knowledge is primarily organised by taxonomy whereas extensive experience results in a more detailed knowledge base in which theme is a powerful organising principle. PMID:22063815

Proper coding of the Abbreviated Injury Scale: can clinical parameters help as surrogates in estimating blood loss?

PubMed

Burkhardt, M; Holstein, J H; Moersdorf, P; Kristen, A; Lefering, R; Pohlemann, T; Pizanis, A

2014-08-01

The Abbreviated Injury Scale (AIS) requires the estimation of the lost blood volume for some severity assignments. This study aimed to develop a rule of thumb for facilitating AIS coding by using objective clinical parameters as surrogate markers of blood loss. Using the example of pelvic ring fractures, a retrospective analysis of TraumaRegister DGU(®) data from 2002 to 2011 was performed. As potential surrogate markers of blood loss, we recorded the hemoglobin (Hb) level, systolic blood pressure (SBP), base excess (BE), Quick's value, units of packed red blood cells (PRBCs) transfused before intensive care unit (ICU) admission, and mortality within 24 h. We identified 11,574 patients with pelvic ring fractures (Tile/OTA classification: 39 % type A, 40 % type B, 21 % type C). Type C fractures were 73.1 % AISpelvis 4 and 26.9 % AISpelvis 5. Type B fractures were 47 % AISpelvis 3, 47 % AISpelvis 4, and 6 % AISpelvis 5. In type C fractures, cut-off values of <7 g/dL Hb, <90 mmHg SBP, <-9 mmol/L BE, <35 % Quick's value, >15 units PRBCs, and death within 24 h had a positive predictive value of 47 % and a sensitivity of 62 % for AISpelvis 5. In type B fractures, these cut-off values had poor sensitivity (48 %) and positive predictive value (11 %) for AISpelvis 5. We failed to develop a rule of thumb for facilitating a proper future AIS coding using the example of pelvic ring fractures. The estimation of blood loss for severity assignment still remains a noteworthy weakness in the AIS coding of traumatic injuries.

Proper poster presentation: a visual and verbal ABC.

PubMed

Wright, V; Moll, J M

1987-08-01

The 58 posters exhibited at the 1985 Annual General Meeting of the British Society for Rheumatology have been analysed for 13 variables considered important in the construction of a good poster. In particular the attributes of information, simplicity and visual attractiveness were studied. The time spent by viewers was also measured for one selected poster each in immunology, biochemistry, therapeutics and clinical medicine. On the basis of this survey, nine recommendations for proper presentation were made.

A Proper-Motion Corrected, Cross-Matched Catalog Of M Dwarfs In SDSS And FIRST

NASA Astrophysics Data System (ADS)

Arai, Erin; West, A. A.; Thyagarajan, N.; Agüeros, M.; Helfand, D.

2011-05-01

We present a preliminary analysis of M dwarfs identified in both the Sloan Digital Sky Survey (SDSS) and the Very Large Array's (VLA) Faint Images of the Radio Sky at Twenty-centimeters survey (FIRST). The presence of magnetic fields is often associated with indirect magnetic activity measurements, such as H-alpha or X-ray emission. Radio emission, in contrast, is directly proportional to the magnetic field strength in addition to being another measure of activity. We search for stellar radio emission by cross-matching the SDSS DR7 M dwarf sample with the FIRST catalog. The SDSS data allow us to examine the spectra of our objects and correlate the magnetic activity (H-alpha) with the magnetic field strength (radio emission). Accurate positions and proper motions are important for obtaining a complete list of overlapping targets. Positions in FIRST and SDSS need to be proper motion corrected in order to ensure unique target matches since nearby M dwarfs can have significant proper motions (up to 1'' per year). Some previous studies have neglected the significance of proper motions in identifying overlapping targets between SDSS and FIRST; we correct for some of these previous oversights. In addition the FIRST data were taken in multiple epochs; individual images need to be proper motion corrected before the images can be co-added. Our cross-match catalog puts important constraints on models of magnetic field generation in low-mass stars in addition to the true habitability of attending planets.

Aquaporin-4 facilitator TGN-073 promotes interstitial fluid circulation within the blood–brain barrier: [17O]H2O JJVCPE MRI study

PubMed Central

Huber, Vincent J.; Igarashi, Hironaka; Ueki, Satoshi; Kwee, Ingrid L.

2018-01-01

The blood–brain barrier (BBB), which imposes significant water permeability restriction, effectively isolates the brain from the systemic circulation. Seemingly paradoxical, the abundance of aquaporin-4 (AQP-4) on the inside of the BBB strongly indicates the presence of unique water dynamics essential for brain function. On the basis of the highly specific localization of AQP-4, namely, astrocyte end feet at the glia limitans externa and pericapillary Virchow–Robin space, we hypothesized that the AQP-4 system serves as an interstitial fluid circulator, moving interstitial fluid from the glia limitans externa to pericapillary Virchow–Robin space to ensure proper glymphatic flow draining into the cerebrospinal fluid. The hypothesis was tested directly using the AQP-4 facilitator TGN-073 developed in our laboratory, and [17O]H2O JJ vicinal coupling proton exchange MRI, a method capable of tracing water molecules delivered into the blood circulation. The results unambiguously showed that facilitation of AQP-4 by TGN-073 increased turnover of interstitial fluid through the system, resulting in a significant reduction in [17O]H2O contents of cortex with normal flux into the cerebrospinal fluid. The study further suggested that in addition to providing the necessary water for proper glymphatic flow, the AQP-4 system produces a water gradient within the interstitial space promoting circulation of interstitial fluid within the BBB. PMID:29481527

Aquaporin-4 facilitator TGN-073 promotes interstitial fluid circulation within the blood-brain barrier: [17O]H2O JJVCPE MRI study.

PubMed

Huber, Vincent J; Igarashi, Hironaka; Ueki, Satoshi; Kwee, Ingrid L; Nakada, Tsutomu

2018-06-13

The blood-brain barrier (BBB), which imposes significant water permeability restriction, effectively isolates the brain from the systemic circulation. Seemingly paradoxical, the abundance of aquaporin-4 (AQP-4) on the inside of the BBB strongly indicates the presence of unique water dynamics essential for brain function. On the basis of the highly specific localization of AQP-4, namely, astrocyte end feet at the glia limitans externa and pericapillary Virchow-Robin space, we hypothesized that the AQP-4 system serves as an interstitial fluid circulator, moving interstitial fluid from the glia limitans externa to pericapillary Virchow-Robin space to ensure proper glymphatic flow draining into the cerebrospinal fluid. The hypothesis was tested directly using the AQP-4 facilitator TGN-073 developed in our laboratory, and [O]H2O JJ vicinal coupling proton exchange MRI, a method capable of tracing water molecules delivered into the blood circulation. The results unambiguously showed that facilitation of AQP-4 by TGN-073 increased turnover of interstitial fluid through the system, resulting in a significant reduction in [O]H2O contents of cortex with normal flux into the cerebrospinal fluid. The study further suggested that in addition to providing the necessary water for proper glymphatic flow, the AQP-4 system produces a water gradient within the interstitial space promoting circulation of interstitial fluid within the BBB.

Radio Measurements of the Stellar Proper Motions in the Core of the Orion Nebula Cluster

NASA Astrophysics Data System (ADS)

Dzib, Sergio A.; Loinard, Laurent; Rodríguez, Luis F.; Gómez, Laura; Forbrich, Jan; Menten, Karl M.; Kounkel, Marina A.; Mioduszewski, Amy J.; Hartmann, Lee; Tobin, John J.; Rivera, Juana L.

2017-01-01

Using multi-epoch Very Large Array observations, covering a time baseline of 29.1 years, we have measured the proper motions of 88 young stars with compact radio emission in the core of the Orion Nebula Cluster (ONC) and the neighboring BN/KL region. Our work increases the number of young stars with measured proper motion at radio frequencies by a factor of 2.5 and enables us to perform a better statistical analysis of the kinematics of the region than was previously possible. Most stars (79 out of 88) have proper motions consistent with a Gaussian distribution centered on \\overline{{μ }α \\cos δ }=1.07+/- 0.09 mas yr-1, and \\overline{{μ }δ }=-0.84+/- 0.16 mas yr-1, with velocity dispersions of {σ }α =1.08+/- 0.07 mas yr-1, {σ }δ =1.27+/- 0.15 mas yr-1. We looked for organized movements of these stars but found no clear indication of radial expansion/contraction or rotation. The remaining nine stars in our sample show peculiar proper motions that differ from the mean proper motions of the ONC by more than 3σ. One of these stars, V 1326 Ori, could have been expelled from the Orion Trapezium 7000 years ago. Two could be related to the multi-stellar disintegration in the BN/KL region, in addition to the previously known sources BN, I and n. The others either have high uncertainties (so their anomalous proper motions are not firmly established) or could be foreground objects.

Proper motion of the Draco dwarf galaxy based on Hubble space telescope imaging

DOE Office of Scientific and Technical Information (OSTI.GOV)

Pryor, Carlton; Piatek, Slawomir; Olszewski, Edward W., E-mail: pryor@physics.rutgers.edu, E-mail: piatek@physics.rutgers.edu, E-mail: eolszewski@as.arizona.edu

2015-02-01

We have measured the proper motion of the Draco dwarf galaxy using images at two epochs with a time baseline of about two years taken with the Hubble Space Telescope Advanced Camera for Surveys. Wide Field Channels 1 and 2 provide two adjacent fields, each containing a known QSO. The zero point for the proper motion is determined using both background galaxies and the QSOs and the two methods produce consistent measurements within each field. Averaging the results from the two fields gives a proper motion in the equatorial coordinate system of (μ{sub α},μ{sub δ})=(17.7±6.3,−22.1±6.3) mas century{sup −1} and inmore » the Galactic coordinate system of (μ{sub ℓ},μ{sub b})=(−23.1±6.3,−16.3±6.3) mas century{sup −1}. Removing the contributions of the motion of the Sun and of the LSR to the measured proper motion yields a Galactic rest-frame proper motion of (μ{sub α}{sup Grf},μ{sub δ}{sup Grf})=(51.4±6.3,−18.7±6.3) mas century{sup −1} and (μ{sub ℓ}{sup Grf},μ{sub b}{sup Grf})=(−21.8±6.3,−50.1±6.3) mas century{sup −1}. The implied space velocity with respect to the Galactic center is (Π,Θ,Z)=(27±14,89±25,−212±20) km s{sup −1}. This velocity implies that the orbital inclination is 70{sup ∘}, with a 95% confidence interval of (59{sup ∘},80{sup ∘}), and that the plane of the orbit is consistent with that of the vast polar structure (VPOS) of Galactic satellite galaxies.« less

UCAC3 Proper Motion Survey. I. Discovery of New Proper Motion Stars in UCAC3 With 0.40/yr mu 0.18/yr Between Declinations -90 deg and -47 deg

DTIC Science & Technology

2010-09-01

overlooked during previous SCR and other searches. The Two-Micron All Sky Survey ( 2MASS ) was used to probe for and reduce systematic errors in UCAC CCD...of 50–200 mas, when compared to 2MASS data. For a detailed description of the derived UCAC3 proper motions see Zacharias et al. (2010). An effort was...meeting the declination and proper motion survey limits, all stars (1) must be in the 2MASS catalog with an e2mpho ( 2MASS photometry error) less than

Testing stellar proper motions of TGAS stars using data from the HSOY, UCAC5 and PMA catalogues

NASA Astrophysics Data System (ADS)

Fedorov, P. N.; Akhmetov, V. S.; Velichko, A. B.

2018-05-01

We analyse the stellar proper motions from the Tycho-Gaia Astrometric Solution (TGAS) and those from the ground-based HSOY, UCAC5 and PMA catalogues derived by combining them with Gaia DR1 space data. Assuming that systematic differences in stellar proper motions of the two catalogues are caused by a mutual rigid-body rotation of the reference catalogue systems, we analyse components of the rotation vector between the systems. We found that the ωy component of the rotation vector is ˜1.5 mas yr-1 and it depends non-linearly on stellar magnitude for the objects of 9.5-11.5 mag used in all three comparisons of the catalogues HSOY, UCAC5 and PMA with respect to TGAS. We found that the Tycho-2 stars in TGAS appeared to have an inexplicable dependence of proper motion on stellar magnitude. We showed that the proper motions of the TGAS stars derived using AGIS differ from those obtained by the conventional (classical) method. Moreover, the application of both methods has not revealed such a difference between the proper motions of the Hipparcos and TGAS stars. An analysis of the systematic differences between the proper motions of the TGAS stars derived by the classical method and the proper motions of the HSOY, UCAC5 and PMA stars shows that the ωy component here does not depend on the magnitude. This indicates unambiguously that there is a magnitude error in the proper motions of the Tycho-2 stars derived with the AGIS.

Facilitating Facilitators to Facilitate, in Problem or Enquiry Based Learning Sessions

ERIC Educational Resources Information Center

Coelho, Catherine

2014-01-01

Problem based learning (PBL) has been used in dental education over the past 20 years and uses a patient case scenario to stimulate learning in a small group setting, where a trained facilitator does not teach but guides the group to bring about deep contextualized learning, to be empathetic to each other and to encourage fair and equitable…

The Optical Gravitational Lensing Experiment. The Catalog of Stellar Proper Motions toward the Magellanic Clouds

NASA Astrophysics Data System (ADS)

Poleski, R.; Soszyński, I.; Udalski, A.; Szymański, M. K.; Kubiak, M.; Pietrzyński, G.; Wyrzykowski, Ł.; Ulaczyk, K.

2012-03-01

We present a catalog of over 6.2 million stars with measured proper motions. All these stars are observed in the direction of the Magellanic Clouds within the brightness range 12proper motions about 440 000 Galactic foreground stars can be selected. Because the proper motions are based on a few hundred epochs collected during eight years, their statistical uncertainties are below 0.5 mas/yr for stars brighter than I=18.5 mag. The parallaxes are derived with uncertainties down to 1.6 mas. For above 13 000 objects parallaxes are derived with significance above 3σ, which allows selecting about 270 white dwarfs (WDs). The search for common proper motion binaries among stars presented was performed resulting in over 500 candidate systems. The most interesting ones are candidate halo main sequence star-WD and WD-WD systems. The application of the catalog to the empirically bound Cepheid instability strip is also discussed.

Golgi Localized Barley MTP8 Proteins Facilitate Mn Transport

PubMed Central

Pedas, Pai; Schiller Stokholm, Michaela; Hegelund, Josefine Nymark; Ladegård, Anne Hald; Schjoerring, Jan Kofod; Husted, Søren

2014-01-01

Many metabolic processes in plants are regulated by manganese (Mn) but limited information is available on the molecular mechanisms controlling cellular Mn homeostasis. In this study, a yeast assay was used to isolate and characterize two genes, MTP8.1 and MTP8.2, which encode membrane-bound proteins belonging to the cation diffusion facilitator (CDF) family in the cereal species barley (Hordeum vulgare). Transient expression in onion epidermal cells showed that MTP8.1 and MTP8.2 proteins fused to the green fluorescent protein (GFP) are localized to Golgi. When heterologously expressed in yeast, MTP8.1 and MTP8.2 were found to be Mn transporters catalysing Mn efflux in a similar manner as the Golgi localized endogenous yeast protein Pmr1p. The level of MTP8.1 transcripts in barley roots increased with external Mn supply ranging from deficiency to toxicity, while MTP8.2 transcripts decreased under the same conditions, indicating non-overlapping functions for the two genes. In barley leaves, the expression of both MTP8 genes declined in response to toxic Mn additions to the roots suggesting a role in ensuring proper delivery of Mn to Golgi. Based on the above we suggest that barley MTP8 proteins are involved in Mn loading to the Golgi apparatus and play a role in Mn homeostasis by delivering Mn to Mn-dependent enzymes and/or by facilitating Mn efflux via secretory vesicles. This study highlights the importance of MTP transporters in Mn homeostasis and is the first report of Golgi localized Mn2+ transport proteins in a monocot plant species. PMID:25486417

Effectiveness of Various Methods of Teaching Proper Inhaler Technique.

PubMed

Axtell, Samantha; Haines, Seena; Fairclough, Jamie

2017-04-01

To compare the effectiveness of 4 different instructional interventions in training proper inhaler technique. Randomized, noncrossover trial. Health fair and indigent clinic. Inhaler-naive adult volunteers who spoke and read English. Subjects were assigned to complete the following: (1) read a metered dose inhaler (MDI) package insert pamphlet, (2) watch a Centers for Disease Control and Prevention (CDC) video demonstrating MDI technique, (3) watch a YouTube video demonstrating MDI technique, or (4) receive direct instruction of MDI technique from a pharmacist. Inhaler use competency (completion of all 7 prespecified critical steps). Of the 72 subjects, 21 (29.2%) demonstrated competent inhaler technique. A statistically significant difference between pharmacist direct instruction and the remaining interventions, both combined ( P < .0001) and individually ( P ≤ .03), was evident. No statistically significant difference was detected among the remaining 3 intervention groups. Critical steps most frequently omitted or improperly performed were exhaling before inhalation and holding of breath after inhalation. A 2-minute pharmacist counseling session is more effective than other interventions in successfully educating patients on proper inhaler technique. Pharmacists can play a pivotal role in reducing the implications of improper inhaler use.

Proper temperature of the Schwarzschild AdS black hole revisited

NASA Astrophysics Data System (ADS)

Eune, Myungseok; Kim, Wontae

2017-10-01

The Unruh temperature calculated by using the global embedding of the Schwarzschild AdS spacetime into the Minkowski spacetime was identified with the local proper temperature; however, it became imaginary in a certain region outside the event horizon. So, the temperature was assumed to be zero of non-thermal radiation for that region. In this work, we revisit this issue in an exactly soluble two-dimensional Schwarzschild AdS black hole and present an alternative resolution to this problem in terms of the Tolman's procedure. However, the process appears to be non-trivial in the sense that the original procedure assuming the traceless energy-momentum tensor should be extended in such a way that it should cover the non-vanishing case of the energy-momentum tensor in the presence of the trace anomaly. Consequently, we show that the proper temperature turns out to be real everywhere outside the event horizon without any imaginary value, in particular, it vanishes at both the horizon and the asymptotic infinity.

Action video game play facilitates the development of better perceptual templates.

PubMed

Bejjanki, Vikranth R; Zhang, Ruyuan; Li, Renjie; Pouget, Alexandre; Green, C Shawn; Lu, Zhong-Lin; Bavelier, Daphne

2014-11-25

The field of perceptual learning has identified changes in perceptual templates as a powerful mechanism mediating the learning of statistical regularities in our environment. By measuring threshold-vs.-contrast curves using an orientation identification task under varying levels of external noise, the perceptual template model (PTM) allows one to disentangle various sources of signal-to-noise changes that can alter performance. We use the PTM approach to elucidate the mechanism that underlies the wide range of improvements noted after action video game play. We show that action video game players make use of improved perceptual templates compared with nonvideo game players, and we confirm a causal role for action video game play in inducing such improvements through a 50-h training study. Then, by adapting a recent neural model to this task, we demonstrate how such improved perceptual templates can arise from reweighting the connectivity between visual areas. Finally, we establish that action gamers do not enter the perceptual task with improved perceptual templates. Instead, although performance in action gamers is initially indistinguishable from that of nongamers, action gamers more rapidly learn the proper template as they experience the task. Taken together, our results establish for the first time to our knowledge the development of enhanced perceptual templates following action game play. Because such an improvement can facilitate the inference of the proper generative model for the task at hand, unlike perceptual learning that is quite specific, it thus elucidates a general learning mechanism that can account for the various behavioral benefits noted after action game play.

Action video game play facilitates the development of better perceptual templates

PubMed Central

Bejjanki, Vikranth R.; Zhang, Ruyuan; Li, Renjie; Pouget, Alexandre; Green, C. Shawn; Lu, Zhong-Lin; Bavelier, Daphne

2014-01-01

The field of perceptual learning has identified changes in perceptual templates as a powerful mechanism mediating the learning of statistical regularities in our environment. By measuring threshold-vs.-contrast curves using an orientation identification task under varying levels of external noise, the perceptual template model (PTM) allows one to disentangle various sources of signal-to-noise changes that can alter performance. We use the PTM approach to elucidate the mechanism that underlies the wide range of improvements noted after action video game play. We show that action video game players make use of improved perceptual templates compared with nonvideo game players, and we confirm a causal role for action video game play in inducing such improvements through a 50-h training study. Then, by adapting a recent neural model to this task, we demonstrate how such improved perceptual templates can arise from reweighting the connectivity between visual areas. Finally, we establish that action gamers do not enter the perceptual task with improved perceptual templates. Instead, although performance in action gamers is initially indistinguishable from that of nongamers, action gamers more rapidly learn the proper template as they experience the task. Taken together, our results establish for the first time to our knowledge the development of enhanced perceptual templates following action game play. Because such an improvement can facilitate the inference of the proper generative model for the task at hand, unlike perceptual learning that is quite specific, it thus elucidates a general learning mechanism that can account for the various behavioral benefits noted after action game play. PMID:25385590

A Comparative Study of the Aneugenic and Polyploidy-inducing Effects of Fisetin and Two Model Aurora Kinase Inhibitors

PubMed Central

Gollapudi, P.; Hasegawa, L.S.; Eastmond, D.A.

2014-01-01

Fisetin, a plant flavonol commonly found in fruits, nuts and vegetables, is frequently added to nutritional supplements due to its reported cardioprotective, anti-carcinogenic and antioxidant properties. Earlier reports from our laboratory and others have indicated that fisetin has both aneugenic and clastogenic properties in cultured cells. More recently, fisetin has also been reported to target Aurora B kinase, a Ser/Thr kinase involved in ensuring proper microtubule attachment at the spindle assembly checkpoint, and an enzyme that is overexpressed in several types of cancer. Here we have further characterized the chromosome damage caused by fisetin and compared it with that induced by two known Aurora kinase inhibitors, VX-680 and ZM-447439, in cultured TK6 cells using the micronucleus assay with CREST staining as well as a flow cytometry-based assay that measures multiple types of numerical chromosomal aberrations. The three compounds were highly effective in inducing aneuploidy and polyploidy as evidenced by increases in kinetochore-positive micronuclei, hyperdiploidy, and polyploidy. With fisetin, however, the latter two effects were most significantly observed only after cells were allowed to overcome a cell cycle delay, and occurred at higher concentrations than those induced by the other Aurora kinase inhibitors. Modest increases in kinetochore-negative micronuclei were also seen with the model Aurora kinase inhibitors. These results indicate that fisetin induces multiple types of chromosome abnormalities in human cells, and indicate a need for a thorough investigation of fisetin-augmented dietary supplements. PMID:24680981

Temporal and SUMO-specific SUMOylation contribute to the dynamics of Polo-like kinase 1 (PLK1) and spindle integrity during mouse oocyte meiosis.

PubMed

Feitosa, Weber Beringui; Hwang, KeumSil; Morris, Patricia L

2018-02-15

During mammalian meiosis, Polo-like kinase 1 (PLK1) is essential during cell cycle progression. In oocyte maturation, PLK1 expression is well characterized but timing of posttranslational modifications regulating its activity and subcellular localization are less clear. Small ubiquitin-related modifier (SUMO) posttranslational modifier proteins have been detected in mammalian gametes but their precise function during gametogenesis is largely unknown. In the present paper we report for mouse oocytes that both PLK1 and phosphorylated PLK1 undergo SUMOylation in meiosis II (MII) oocytes using immunocytochemistry, immunoprecipitation and in vitro SUMOylation assays. At MII, PLK1 is phosphorylated at threonine-210 and serine-137. MII oocyte PLK1 and phosphorylated PLK1 undergo SUMOylation by SUMO-1, -2 and -3 as shown by individual in vitro assays. Using these assays, forms of phosphorylated PLK1 normalized to PLK1 increased significantly and correlated with SUMOylated PLK1 levels. During meiotic progression and maturation, SUMO-1-SUMOylation of PLK1 is involved in spindle formation whereas SUMO-2/3-SUMOylation may regulate PLK1 activity at kinetochore-spindle attachment sites. Microtubule integrity is required for PLK1 localization with SUMO-1 but not with SUMO-2/3. Inhibition of SUMOylation disrupts proper meiotic bipolar spindle organization and spindle-kinetochore attachment. The data show that both temporal and SUMO-specific-SUMOylation play important roles in orchestrating functional dynamics of PLK1 during mouse oocyte meiosis, including subcellular compartmentalization. Copyright © 2018 Elsevier Inc. All rights reserved.

Regulation of Centromere Localization of the Drosophila Shugoshin MEI-S332 and Sister-Chromatid Cohesion in Meiosis

PubMed Central

Nogueira, Cristina; Kashevsky, Helena; Pinto, Belinda; Clarke, Astrid; Orr-Weaver, Terry L.

2014-01-01

The Shugoshin (Sgo) protein family helps to ensure proper chromosome segregation by protecting cohesion at the centromere by preventing cleavage of the cohesin complex. Some Sgo proteins also influence other aspects of kinetochore-microtubule attachments. Although many Sgo members require Aurora B kinase to localize to the centromere, factors controlling delocalization are poorly understood and diverse. Moreover, it is not clear how Sgo function is inactivated and whether this is distinct from delocalization. We investigated these questions in Drosophila melanogaster, an organism with superb chromosome cytology to monitor Sgo localization and quantitative assays to test its function in sister-chromatid segregation in meiosis. Previous research showed that in mitosis in cell culture, phosphorylation of the Drosophila Sgo, MEI-S332, by Aurora B promotes centromere localization, whereas Polo phosphorylation promotes delocalization. These studies also suggested that MEI-S332 can be inactivated independently of delocalization, a conclusion supported here by localization and function studies in meiosis. Phosphoresistant and phosphomimetic mutants for the Aurora B and Polo phosphorylation sites were examined for effects on MEI-S332 localization and chromosome segregation in meiosis. Strikingly, MEI-S332 with a phosphomimetic mutation in the Aurora B phosphorylation site prematurely dissociates from the centromeres in meiosis I. Despite the absence of MEI-S332 on meiosis II centromeres in male meiosis, sister chromatids segregate normally, demonstrating that detectable levels of this Sgo are not essential for chromosome congression, kinetochore biorientation, or spindle assembly. PMID:25081981

Attention to baseline: does orienting visuospatial attention really facilitate target detection?

PubMed

Albares, Marion; Criaud, Marion; Wardak, Claire; Nguyen, Song Chi Trung; Ben Hamed, Suliann; Boulinguez, Philippe

2011-08-01

Standard protocols testing the orientation of visuospatial attention usually present spatial cues before targets and compare valid-cue trials with invalid-cue trials. The valid/invalid contrast results in a relative behavioral or physiological difference that is generally interpreted as a benefit of attention orientation. However, growing evidence suggests that inhibitory control of response is closely involved in this kind of protocol that requires the subjects to withhold automatic responses to cues, probably biasing behavioral and physiological baselines. Here, we used two experiments to disentangle the inhibitory control of automatic responses from orienting of visuospatial attention in a saccadic reaction time task in humans, a variant of the classical cue-target detection task and a sustained visuospatial attentional task. Surprisingly, when referring to a simple target detection task in which there is no need to refrain from reacting to avoid inappropriate responses, we found no consistent evidence of facilitation of target detection at the attended location. Instead, we observed a cost at the unattended location. Departing from the classical view, our results suggest that reaction time measures of visuospatial attention probably relie on the attenuation of elementary processes involved in visual target detection and saccade initiation away from the attended location rather than on facilitation at the attended location. This highlights the need to use proper control conditions in experimental designs to disambiguate relative from absolute cueing benefits on target detection reaction times, both in psychophysical and neurophysiological studies.

5 CFR 550.808 - Prohibition against setting aside proper promotions.

Code of Federal Regulations, 2010 CFR

2010-01-01

... promotions. 550.808 Section 550.808 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE... promotions. Nothing in section 5596 of title 5, United States Code, or this subpart shall be construed as authorizing the setting aside of an otherwise proper promotion by a selecting official from a group of...

Localization of spindle checkpoint proteins in cells undergoing mitosis with unreplicated genomes.

PubMed

Johnson, Mary Kathrine; Cooksey, Amanda M; Wise, Dwayne A

2008-11-01

CHO cells can be arrested with hydoxyurea at the beginning of the DNA synthesis phase of the cell cycle. Subsequent treatment with the xanthine, caffeine, induces cells to bypass the S-phase checkpoint and enter unscheduled mitosis [Schlegel and Pardee,1986, Science 232:1264-1266]. These treated cells build a normal spindle and distribute kinetochores, unattached to chromosomes, to their daughter cells [Brinkley et al.,1988, Nature 336:251-254; Zinkowski et al.,1991, J Cell Biol 113:1091-1110; Wise and Brinkley,1997, Cell Motil Cytoskeleton 36:291-302; Balczon et al.,2003, Chromosoma 112:96-102]. To investigate how these cells distribute kinetochores to daughter cells, we analyzed the spindle checkpoint components, Mad2, CENP-E, and the 3F3 phosphoepitope, using immunofluorescence and digital microscopy. Even though the kinetochores were unpaired and DNA was fragmented, the tension, alignment, and motor components of the checkpoint were found to be present and localized as predicted in prometaphase and metaphase. This unusual mitosis proves that a cell can successfully localize checkpoint proteins and divide even when kinetochores are unpaired and fragmented. (c) 2008 Wiley-Liss, Inc.

A sequential multi-target Mps1 phosphorylation cascade promotes spindle checkpoint signaling.

PubMed

Ji, Zhejian; Gao, Haishan; Jia, Luying; Li, Bing; Yu, Hongtao

2017-01-10

The master spindle checkpoint kinase Mps1 senses kinetochore-microtubule attachment and promotes checkpoint signaling to ensure accurate chromosome segregation. The kinetochore scaffold Knl1, when phosphorylated by Mps1, recruits checkpoint complexes Bub1-Bub3 and BubR1-Bub3 to unattached kinetochores. Active checkpoint signaling ultimately enhances the assembly of the mitotic checkpoint complex (MCC) consisting of BubR1-Bub3, Mad2, and Cdc20, which inhibits the anaphase-promoting complex or cyclosome bound to Cdc20 (APC/C Cdc20 ) to delay anaphase onset. Using in vitro reconstitution, we show that Mps1 promotes APC/C inhibition by MCC components through phosphorylating Bub1 and Mad1. Phosphorylated Bub1 binds to Mad1-Mad2. Phosphorylated Mad1 directly interacts with Cdc20. Mutations of Mps1 phosphorylation sites in Bub1 or Mad1 abrogate the spindle checkpoint in human cells. Therefore, Mps1 promotes checkpoint activation through sequentially phosphorylating Knl1, Bub1, and Mad1. This sequential multi-target phosphorylation cascade makes the checkpoint highly responsive to Mps1 and to kinetochore-microtubule attachment.

Natural Loss of Mps1 Kinase in Nematodes Uncovers a Role for Polo-like Kinase 1 in Spindle Checkpoint Initiation.

PubMed

Espeut, Julien; Lara-Gonzalez, Pablo; Sassine, Mélanie; Shiau, Andrew K; Desai, Arshad; Abrieu, Ariane

2015-07-07

The spindle checkpoint safeguards against chromosome loss during cell division by preventing anaphase onset until all chromosomes are attached to spindle microtubules. Checkpoint signal is generated at kinetochores, the primary attachment site on chromosomes for spindle microtubules. Mps1 kinase initiates checkpoint signaling by phosphorylating the kinetochore-localized scaffold protein Knl1 to create phospho-docking sites for Bub1/Bub3. Mps1 is widely conserved but is surprisingly absent in many nematode species. Here, we show that PLK-1, which targets a substrate motif similar to that of Mps1, functionally substitutes for Mps1 in C. elegans by phosphorylating KNL-1 to direct BUB-1/BUB-3 kinetochore recruitment. This finding led us to re-examine checkpoint initiation in human cells, where we found that Plk1 co-inhibition significantly reduced Knl1 phosphorylation and Bub1 kinetochore recruitment relative to Mps1 inhibition alone. Thus, the finding that PLK-1 functionally substitutes for Mps1 in checkpoint initiation in C. elegans uncovered a role for Plk1 in species that have Mps1. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

A Proper Motion Search for Stars Escaping from Globular Clusters with High Velocities

NASA Astrophysics Data System (ADS)

Meusinger, H.; Scholz, R.-D.; Irwin, M.

The dynamical evolution of globular clusters, in particular during the late phases, may be strongly influenced by the energy transfer from binaries to passing stars. As a by-product of this process, stars with high velocities are expected, perhaps high enough to escape from the cluster. Accurate proper motions are the only suitable tool to identify candidates for such high-velocity cluster stars. In order to perform such a search, we use a catalogue of absolute proper motions and UBV magnitudes for about 104 stars with B < 20 in a field of 10 square degrees centered on the globular cluster M3. The data were derived from more than 80 photographic plates taken between 1965 and 1995 with the Tautenburg Schmidt telescope and measured by means of the APM facility, Cambridge. The stellar sample is complete to B = 18.5 and comprises nearly all post-main-sequence stars in the halo of M3 and its surrounding. The proper motions are of Hipparcos-like accuracy (median error 1 mas/yr) in this magnitude range. We find several dozens of candidates, distributed over the whole field, with proper motions and colours consistent with the assumption of their origin from the cluster. Further conclusions can be drawn only on the basis of radial velocity measurements for the candidates and of estimates for the field-star contamination by means of simulations of the Galactic structure and kinematics in this field.

Proper body mechanics from an engineering perspective.

PubMed

Mohr, Edward G

2010-04-01

The economic viability of the manual therapy practitioner depends on the number of massages/treatments that can be given in a day or week. Fatigue or injuries can have a major impact on the income potential and could ultimately reach the point which causes the practitioner to quit the profession, and seek other, less physically demanding, employment. Manual therapy practitioners in general, and massage therapists in particular, can utilize a large variety of body postures while giving treatment to a client. The hypothesis of this paper is that there is an optimal method for applying force to the client, which maximizes the benefit to the client, and at the same time minimizes the strain and effort required by the practitioner. Two methods were used to quantifiably determine the effect of using "poor" body mechanics (Improper method) and "best" body mechanics (Proper/correct method). The first approach uses computer modeling to compare the two methods. Both postures were modeled, such that the biomechanical effects on the practitioner's elbow, shoulder, hip, knee and ankle joints could be calculated. The force applied to the client, along with the height and angle of application of the force, was held constant for the comparison. The second approach was a field study of massage practitioners (n=18) to determine their maximal force capability, again comparing methods using "Improper and Proper body mechanics". Five application methods were tested at three different application heights, using a digital palm force gauge. Results showed that there was a definite difference between the two methods, and that the use of correct body mechanics can have a large impact on the health and well being of the massage practitioner over both the short and long term. Copyright 2009 Elsevier Ltd. All rights reserved.

TAO1 kinase maintains chromosomal stability by facilitating proper congression of chromosomes

PubMed Central

Shrestha, Roshan L.; Tamura, Naoka; Fries, Anna; Levin, Nicolas; Clark, Joanna; Draviam, Viji M.

2014-01-01

Chromosomal instability can arise from defects in chromosome–microtubule attachment. Using a variety of drug treatments, we show that TAO1 kinase is required for ensuring the normal congression of chromosomes. Depletion of TAO1 reduces the density of growing interphase and mitotic microtubules in human cells, showing TAO1's role in controlling microtubule dynamics. We demonstrate the aneugenic nature of chromosome–microtubule attachment defects in TAO1-depleted cells using an error-correction assay. Our model further strengthens the emerging paradigm that microtubule regulatory pathways are important for resolving erroneous kinetochore–microtubule attachments and maintaining the integrity of the genome, regardless of the spindle checkpoint status. PMID:24898139

Network community-detection enhancement by proper weighting

NASA Astrophysics Data System (ADS)

Khadivi, Alireza; Ajdari Rad, Ali; Hasler, Martin

2011-04-01

In this paper, we show how proper assignment of weights to the edges of a complex network can enhance the detection of communities and how it can circumvent the resolution limit and the extreme degeneracy problems associated with modularity. Our general weighting scheme takes advantage of graph theoretic measures and it introduces two heuristics for tuning its parameters. We use this weighting as a preprocessing step for the greedy modularity optimization algorithm of Newman to improve its performance. The result of the experiments of our approach on computer-generated and real-world data networks confirm that the proposed approach not only mitigates the problems of modularity but also improves the modularity optimization.

Developing facilitation skills--a narrative.

PubMed

Newton, Jennifer M

2003-07-01

Effective facilitation has been identified in the literature as one of three elements, along with context and evidence, that have a dynamic and coexisting relationship to enable the successful uptake of evidence into practice. This paper presents an overview of the concept of facilitation within the context of practice development, ahead of a personal and professional reflective account of a 'developing facilitator'. In the summer of 2001, the author was instrumental in organising the first Practice Development School in Melbourne. Thrown in at the deep end, she found herself co-facilitating with an experienced practice developer from the United Kingdom. Having never facilitated in the arena of an action learning group, nor worked in the field of practice development, there was initially a sense of impending overload and drowning in the new knowledge and skills that needed to be acquired. Drawing upon the work of narrative inquiry the author shares her experiences in the anticipation that in telling her story it will assist others in their journey of becoming a facilitator.

A Case of Ruptured Aneurysm of the Proper Esophageal Artery with Symptomatic Mediastinal Hematoma

DOE Office of Scientific and Technical Information (OSTI.GOV)

Liu, Jiajia; Sato, Yusuke, E-mail: yusuke@doc.med.akita-u.ac.jp; Takahashi, Satoshi

Mediastinal aneurysms are rare but potentially life-threatening. Among these, bronchial artery aneurysms are most frequently reported, whereas up to now aneurysms of the proper esophageal artery had never been reported. A 69-year-old woman was referred to our hospital for treatment of a massive mediastinal hematoma. Enhanced computed tomography and selective proper esophageal arteriography revealed a 5-mm aneurysm in the proper esophageal artery that arises from the thoracic aorta at the Th8 level and has an anastomotic branch with the bronchial artery peripherally. Transcatheter arterial embolization was successfully performed using a mixture of N-butyl cyanoacrylate and lipiodol (1:3 ratio, 0.3 ml). Post-embolizationmore » angiography showed no filling into the aneurysm. The patient recovered with no complications and was discharged on the 25th post-procedure day.« less

Medical students' and facilitators' experiences of an Early Professional Contact course: active and motivated students, strained facilitators.

PubMed

von Below, Bernhard; Hellquist, Gunilla; Rödjer, Stig; Gunnarsson, Ronny; Björkelund, Cecilia; Wahlqvist, Mats

2008-12-02

Today, medical students are introduced to patient contact, communication skills, and clinical examination in the preclinical years of the curriculum with the purpose of gaining clinical experience. These courses are often evaluated from the student perspective. Reports with an additional emphasis on the facilitator perspective are scarce. According to constructive alignment, an influential concept from research in higher education, the learning climate between students and teachers is also of great importance. In this paper, we approach the learning climate by studying both students' and facilitators' course experiences.In 2001, a new "Early Professional Contact" longitudinal strand through term 1-4, was introduced at the Sahlgrenska Academy, University of Gothenburg, Sweden. General practitioners and hospital specialists were facilitators.The aim of this study was to assess and analyse students' and clinical facilitators' experiences of the Early Professional Contact course and to illuminate facilitators' working conditions. Inspired by a Swedish adaptation of the Course Experience Questionnaire, an Early Professional Contact Questionnaire was constructed. In 2003, on the completion of the first longitudinal strand, a student and facilitator version was distributed to 86 students and 21 facilitators. In the analysis, both Chi-square and the Mann-Whitney tests were used. Sixty students (70%) and 15 facilitators (71%) completed the questionnaire. Both students and facilitators were satisfied with the course. Students reported gaining iiration for their future work as doctors along with increased confidence in meeting patients. They also reported increased motivation for biomedical studies. Differences in attitudes between facilitators and students were found. Facilitators experienced a greater workload, less reasonable demands and less support, than students. In this project, a new Early Professional Contact course was analysed from both student and facilitator

DOE Office of Scientific and Technical Information (OSTI.GOV)

Vladimir Larionov, Ph D

A special interest in the organization of human centromeric DNA was stimulated a few years ago when two independent groups succeeded in reconstituting a functional human centromere, using constructs carrying centromere-specific alphoid DNA arrays. This work demonstrated the importance of DNA components in mammalian centromeres and opened a way for studying the structural requirements for de novo kinetochore formation and for construction of human artificial chromosomes (HACs) with therapeutic potential. To elucidate the structural requirements for formation of HACs with a functional kinetochore, we developed a new method for cloning of large DNA fragments for human centromeric regions that canmore » be used as a substrate for HAC formation. This method exploits in vivo recombination in yeast (TAR cloning). In addition, a new strategy for the construction of alphoid DNA arrays was developed in our lab. The strategy involves the construction of uniform or hybrid synthetic alphoid DNA arrays by the RCA-TAR technique. This technique comprises two steps: rolling circle amplification of an alphoid DNA dimer and subsequent assembling of the amplified fragments by in vivo homologous recombination in yeast (Figure 1). Using this system, we constructed a set of different synthetic alphoid DNA arrays with a predetermined sequence varying in size from 30 to 140 kb and demonstrated that some of the arrays are competent in HAC formation. Because any nucleotide can be changed in a dimer before its amplification, this new technique is optimal for identifying the structural requirements for de novo kinetochore formation in HACs. Moreover, the technique makes possible to introduce into alphoid DNA arrays recognition sites for DNA-binding proteins. We have made the following progress on the studying of human centromeric regions using transformation-associated recombination cloning technology: i) minimal size of alphoid DNA array required for de novo kinetochore formation was

CASPASE-9 CARD:CORE DOMAIN INTERACTIONS REQUIRE A PROPERLY-FORMED ACTIVE SITE

PubMed Central

Huber, Kristen L.; Serrano, Banyuhay P.; Hardy, Jeanne A.

2018-01-01

Caspase-9 is a critical factor in the initiation of apoptosis, and as a result is tightly regulated by a number of mechanisms. Caspase-9 contains a Caspase Activation and Recruitment Domain (CARD), which enables caspase-9 to form a tight interaction with the apoptosome, a heptameric activating platform. The caspase-9 CARD has been thought to be principally involved in recruitment to the apoptosome, but its roles outside this interaction have yet to be uncovered. In this work we show that the CARD is involved in physical interactions with the catalytic core of caspase-9 in the absence of the apoptosome; this interaction requires a properly formed caspase-9 active site. The active sites of caspases are composed of four extremely mobile loops. When the active-site loops are not properly ordered, the CARD and core domains of caspase-9 do not interact and behave independently, like loosely tethered beads. When the active-site loop bundle is properly ordered, the CARD domain interacts with the catalytic core, forming a single folding unit. Together these findings provide mechanistic insight into a new level of caspase-9 regulation, prompting speculation that the CARD may also play a role in the recruitment or recognition of substrate. PMID:29500231

VizieR Online Data Catalog: Proper motions in M 11 (Su+ 1998)

NASA Astrophysics Data System (ADS)

Su, C.-G.; Zhao, J.-L.; Tian, K.-P.

1997-07-01

Relative proper motions of 872 stars in the open cluster M 11 region are reduced using 10 plate pairs taken over time baselines of 16~70 years with the double astrograph telescope of Shanghai Observatory. The scale is 30"/mm. The plates were measured with the PDS machines in the Purple Mountain Observatory in Nanjing and the Institute of Technology and Communication in Luoyang, China. The average proper motion accuracy is about 1.1mas/yr with 85% of the data better than 1mas/yr. Membership probabilities of 785 stars within 25' centred on M 11 are determined based on their proper motions. The method used is suggested by Su et al. (1995AcApS..15..217S) with some improvements of Zhao & He (1990A&A...237...54Z), in which the space distribution and magnitude dependencies for cluster stars are taken into account. The results are significantly good. The total integrated membership probabilities for all these stars is 547 and the number of stars with probabilities higher than 0.7 is 541. It can be found after the membership determination that there exists mass segregation in M 11. Some comparisons and discussion are also given. (1 data file).

Computing Role Assignments of Proper Interval Graphs in Polynomial Time

NASA Astrophysics Data System (ADS)

Heggernes, Pinar; van't Hof, Pim; Paulusma, Daniël

A homomorphism from a graph G to a graph R is locally surjective if its restriction to the neighborhood of each vertex of G is surjective. Such a homomorphism is also called an R-role assignment of G. Role assignments have applications in distributed computing, social network theory, and topological graph theory. The Role Assignment problem has as input a pair of graphs (G,R) and asks whether G has an R-role assignment. This problem is NP-complete already on input pairs (G,R) where R is a path on three vertices. So far, the only known non-trivial tractable case consists of input pairs (G,R) where G is a tree. We present a polynomial time algorithm that solves Role Assignment on all input pairs (G,R) where G is a proper interval graph. Thus we identify the first graph class other than trees on which the problem is tractable. As a complementary result, we show that the problem is Graph Isomorphism-hard on chordal graphs, a superclass of proper interval graphs and trees.

A Spectroscopic Catalog of Nearby, High Proper Motion M subdwarfs

NASA Astrophysics Data System (ADS)

Hejazi, Neda; Lepine, Sebastien; Homeier, Derek

2018-01-01

We present a catalog of 350 metal-poor M subdwarfs, most of them likely from the local Galactic halo population, assembled from medium-resolution observations made at the MDM observatory. All objects are high proper motion stars, with 257 of them having proper motions > 0.4"/yr. We have identified the brightest prototypes for each bin of a grid of 14 spectral subtypes (M0, M0.5, M1, … M6.5) and 9 metallicity bins that go from the moderately metal-poor subdwarfs (sdM), to the more metal-poor extreme subdwarfs (esdM), to the most metal-poor ultra subdwarfs (usdM), each of which is subdivided into three finer metallicity subclasses. The spectral classification by subtype and metallicity class has been determined by a template-fit method, and confirmed by synthetic-model fitting using the BT-Settl spectral grid. We provide the list of the brightest prototypes for each subtype/subclass, as a guide for future high-resolution surveys of low-mass, metal-poor stars.

Monopolar spindle 1 (MPS1) kinase promotes production of closed MAD2 (C-MAD2) conformer and assembly of the mitotic checkpoint complex.

PubMed

Tipton, Aaron R; Ji, Wenbin; Sturt-Gillespie, Brianne; Bekier, Michael E; Wang, Kexi; Taylor, William R; Liu, Song-Tao

2013-12-06

MPS1 kinase is an essential component of the spindle assembly checkpoint (SAC), but its functioning mechanisms are not fully understood. We have shown recently that direct interaction between BUBR1 and MAD2 is critical for assembly and function of the human mitotic checkpoint complex (MCC), the SAC effector. Here we report that inhibition of MPS1 kinase activity by reversine disrupts BUBR1-MAD2 as well as CDC20-MAD2 interactions, causing premature activation of the anaphase-promoting complex/cyclosome. The effect of MPS1 inhibition is likely due to reduction of closed MAD2 (C-MAD2), as expressing a MAD2 mutant (MAD2(L13A)) that is locked in the C conformation rescued the checkpoint defects. In the presence of reversine, exogenous C-MAD2 does not localize to unattached kinetochores but is still incorporated into the MCC. Contrary to a previous report, we found that sustained MPS1 activity is required for maintaining both the MAD1·C-MAD2 complex and open MAD2 (O-MAD2) at unattached kinetochores to facilitate C-MAD2 production. Additionally, mitotic phosphorylation of BUBR1 is also affected by MPS1 inhibition but seems dispensable for MCC assembly. Our results support the notion that MPS1 kinase promotes C-MAD2 production and subsequent MCC assembly to activate the SAC.

Breathe easy with proper respiratory protection

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bidwell, J.

1996-05-01

Evaluating the need for respiratory protection in chemical process industries (CPI) plants and selecting the appropriate respirator involves several steps. The Occupational Safety and Health Administration (OSHA) general industry standard for respiratory protection (29 CFR 1910.134(b)) requires the employer to establish a program to help reduce exposures to occupational contaminants. When feasible, employers must eliminate contaminants by using engineering controls (such as general and local ventilation, enclosure or isolation, or substitution of a less-hazardous process or material). Establishing a respiratory protection program consists of four steps: (1) Identify respiratory hazards and concentrations; (2) Understand the contaminants` effects on workers` health;more » (3) Select appropriate respiratory protection; and (4) Train in proper respirator use and maintenance. Consult applicable state and OSHA requirements to ensure that your program satisfies these steps. Industrial respirator manufacturers can assist with on-site training and fit testing. The paper discusses these four steps, program guidelines, determination of the hazard, and styles of respirators.« less

Online interprofessional education facilitation: A scoping review.

PubMed

Evans, Sherryn Maree; Ward, Catherine; Reeves, Scott

2018-04-22

The use of online media to deliver interprofessional education (IPE) is becoming more prevalent across health professions education settings. Facilitation of IPE activities is known to be critical to the effective delivery of IPE, however, specifics about the nature of online IPE facilitation remains unclear. To explore the health professions education literature to understand the extent, range and nature of research on online IPE facilitation. Scoping review methodology was used to guide a search of four electronic databases for relevant papers. Of the 2095 abstracts initially identified, after screening of both abstracts and full-text papers, 10 studies were selected for inclusion in this review. Following abstraction of key information from each study, a thematic analysis was undertaken. Three key themes emerged to describe the nature of the IPE facilitation literature: (1) types of online IPE facilitation contributions, (2) the experience of online IPE facilitation and (3) personal outcomes of online IPE facilitation. These IPE facilitation themes were particularly focused on facilitation of interprofessional student teams on an asynchronous basis. While the included studies provide some insight into the nature of online IPE facilitation, future research is needed to better understand facilitator contributions, and the facilitation experience and associated outcomes, both relating to synchronous and asynchronous online environments.

Functional Characterization of CENP-A Post-Translational Modifications in Chromosome Segregation

DTIC Science & Technology

2016-09-01

our overall findings in discussion part, and finally we will explain major materials and methods we used. Results CENP-A α-amino methylation...centromere and kinetochore and accurate segregation of the genetic materials . Moreover, we established that centromere/kinetochore defects in the absence...developed. Materials and methods: Creation of CENP-A complete replacement RPE cells: RPE CENP-A knockout cell line generated by Don Cleaveland Lab7 used

UCAC3 PROPER MOTION SURVEY. I. DISCOVERY OF NEW PROPER MOTION STARS IN UCAC3 WITH 0.''40 yr{sup -1} > {mu} {>=} 0.''18 yr{sup -1} BETWEEN DECLINATIONS -90{sup 0} AND -47{sup 0}

DOE Office of Scientific and Technical Information (OSTI.GOV)

Finch, Charlie T.; Zacharias, Norbert; Henry, Todd J., E-mail: finch@usno.navy.mi

2010-09-15

This paper presents 442 new proper motion stellar systems in the southern sky between declinations -90{sup 0} and -47{sup 0} with 0.''40 yr{sup -1} > {mu} {>=} 0.''18 yr{sup -1}. These systems constitute a 25.3% increase in new systems for the same region of the sky covered by previous SuperCOSMOS RECONS (SCR) searches that used Schmidt plates as the primary source of discovery. Among the new systems are 25 multiples, plus an additional 7 new common proper motion (CPM) companions to previously known primaries. All stars have been discovered using the third U.S. Naval Observatory (USNO) CCD Astrograph Catalog (UCAC3).more » A comparison of the UCAC3 proper motions to those from the Hipparcos, Tycho-2, Southern Proper Motion (SPM4), and SuperCOSMOS efforts is presented and shows that UCAC3 provides similar values and precision to the first three surveys. The comparison between UCAC3 and SuperCOSMOS indicates that proper motions in R.A. are systematically shifted in the SuperCOSMOS data but are consistent in decl. data, while overall showing a significantly higher scatter. Distance estimates are derived for stars having SuperCOSMOS Sky Survey B{sub J} , R{sub 59F}, and I{sub IVN} plate magnitudes and Two-Micron All Sky Survey infrared photometry. We find 15 systems estimated to be within 25 pc, including UPM 1710-5300 our closest new discovery estimated at 13.5 pc. Such new discoveries suggest that more nearby stars are yet to be found in these slower proper motion regimes, indicating that more work is needed to develop a complete map of the solar neighborhood.« less

Proper survey methods for research of aquatic plant ecology and management

USDA-ARS?s Scientific Manuscript database

Proper survey methods are essential for objective, quantitative assessment of the distribution and abundance of aquatic plants as part of research and demonstration efforts. For research, the use of the appropriate method is an essential part of the scientific method, to ensure that the experimenta...

Barriers and Facilitators to Self-Directed Learning in Continuing Professional Development for Physicians in Canada: A Scoping Review.

PubMed

Jeong, Dahn; Presseau, Justin; ElChamaa, Rima; Naumann, Danielle N; Mascaro, Colin; Luconi, Francesca; Smith, Karen M; Kitto, Simon

2018-04-10

This scoping review explored the barriers and facilitators that influence engagement in and implementation of self-directed learning (SDL) in continuing professional development (CPD) for physicians in Canada. This review followed the six-stage scoping review framework of Arksey and O'Malley and of Daudt et al. In 2015, the authors searched eight online databases for English-language Canadian articles published January 2005-December 2015. To chart and analyze the data from the 17 included studies, they employed two-step analysis process of conventional content analysis followed by directed coding guided by the Theoretical Domains Framework (TDF). Conventional content analysis generated five categories of barriers and facilitators: individual, program, technological, environmental, and workplace/organizational. Directed coding guided by the TDF allowed analysis of barriers and facilitators to behavior change according to two key groups: physicians engaging in SDL and SDL developers designing and implementing SDL programs. Of the 318 total barriers and facilitators coded, 290 (91.2%) were coded for physicians and 28 (8.8%) for SDL developers. The majority (209; 65.7%) were coded in four key TDF domains: environmental context and resources, social influences, beliefs about consequences, and behavioral regulation. This scoping review identified five categories of barriers and facilitators in the literature and four key TDF domains where most factors related to behavior change of physicians and SDL developers regarding SDL programs in CPD were coded. There was a significant gap in the literature about factors that may contribute to SDL developers' capacity to design and implement SDL programs in CPD.This is an open-access article distributed under the terms of the Creative Commons Attribution-Non Commercial-No Derivatives License 4.0 (CCBY-NC-ND), where it is permissible to download and share the work provided it is properly cited. The work cannot be changed in any way

Impact of quasar proper motions on the alignment between the International Celestial Reference Frame and the Gaia reference frame

NASA Astrophysics Data System (ADS)

Liu, J.-C.; Malkin, Z.; Zhu, Z.

2018-03-01

The International Celestial Reference Frame (ICRF) is currently realized by the very long baseline interferometry (VLBI) observations of extragalactic sources with the zero proper motion assumption, while Gaia will observe proper motions of these distant and faint objects to an accuracy of tens of microarcseconds per year. This paper investigates the difference between VLBI and Gaia quasar proper motions and it aims to understand the impact of quasar proper motions on the alignment of the ICRF and Gaia reference frame. We use the latest time series data of source coordinates from the International VLBI Service analysis centres operated at Goddard Space Flight Center (GSF2017) and Paris observatory (OPA2017), as well as the Gaia auxiliary quasar solution containing 2191 high-probability optical counterparts of the ICRF2 sources. The linear proper motions in right ascension and declination of VLBI sources are derived by least-squares fits while the proper motions for Gaia sources are simulated taking into account the acceleration of the Solar system barycentre and realistic uncertainties depending on the source brightness. The individual and global features of source proper motions in GSF2017 and OPA2017 VLBI data are found to be inconsistent, which may result from differences in VLBI observations, data reduction and analysis. A comparison of the VLBI and Gaia proper motions shows that the accuracies of the components of rotation and glide between the two systems are 2-4 μas yr- 1 based on about 600 common sources. For the future alignment of the ICRF and Gaia reference frames at different wavelengths, the proper motions of quasars must necessarily be considered.

The Resourceful Facilitator: Teacher Leaders Constructing Identities as Facilitators of Teacher Peer Groups

ERIC Educational Resources Information Center

Allen, David

2016-01-01

The use of teacher peer groups is a prevalent strategy for school-based professional development and instructional improvement. Facilitation of such groups is an increasingly vital dimension of teacher leadership as a component of school improvement efforts. Drawing on a qualitative study of facilitation of teacher peer groups, the article…

Enthalpy Costs of Making and Breaking Bonds: A Game of Generating Molecules with Proper Lewis Structures

ERIC Educational Resources Information Center

Bell, Peter T.; Adkins, Alyssa D.; Gamble, Rex J.; Schultz, Linda D.

2009-01-01

"Enthalpy Costs" is a simple card game created to assist students in developing proper Lewis structure drawing skills. Score keeping is accomplished by tracking the enthalpy changes associated with bond-making and bond-breaking processes during formation of molecules represented by proper Lewis structures. Playing the game requires the student to…

Report: EPA Did Not Properly Process a Hospital Disinfectant and Sanitizer Registration

EPA Pesticide Factsheets

Report #2007-P-00018, March 29, 2007. EPA’s Office of Pesticides Program-Antimicrobials Division (OPP-AD) did not properly process registration for an antimicrobial pesticide that was the subject of our review.

Characterization of Spindle Checkpoint Kinase Mps1 Reveals Domain with Functional and Structural Similarities to Tetratricopeptide Repeat Motifs of Bub1 and BubR1 Checkpoint Kinases*

PubMed Central

Lee, Semin; Thebault, Philippe; Freschi, Luca; Beaufils, Sylvie; Blundell, Tom L.; Landry, Christian R.; Bolanos-Garcia, Victor M.; Elowe, Sabine

2012-01-01

Kinetochore targeting of the mitotic kinases Bub1, BubR1, and Mps1 has been implicated in efficient execution of their functions in the spindle checkpoint, the self-monitoring system of the eukaryotic cell cycle that ensures chromosome segregation occurs with high fidelity. In all three kinases, kinetochore docking is mediated by the N-terminal region of the protein. Deletions within this region result in checkpoint failure and chromosome segregation defects. Here, we use an interdisciplinary approach that includes biophysical, biochemical, cell biological, and bioinformatics methods to study the N-terminal region of human Mps1. We report the identification of a tandem repeat of the tetratricopeptide repeat (TPR) motif in the N-terminal kinetochore binding region of Mps1, with close homology to the tandem TPR motif of Bub1 and BubR1. Phylogenetic analysis indicates that TPR Mps1 was acquired after the split between deutorostomes and protostomes, as it is distinguishable in chordates and echinoderms. Overexpression of TPR Mps1 resulted in decreased efficiency of both chromosome alignment and mitotic arrest, likely through displacement of endogenous Mps1 from the kinetochore and decreased Mps1 catalytic activity. Taken together, our multidisciplinary strategy provides new insights into the evolution, structural organization, and function of Mps1 N-terminal region. PMID:22187426

A sequential multi-target Mps1 phosphorylation cascade promotes spindle checkpoint signaling

PubMed Central

Ji, Zhejian; Gao, Haishan; Jia, Luying; Li, Bing; Yu, Hongtao

2017-01-01

The master spindle checkpoint kinase Mps1 senses kinetochore-microtubule attachment and promotes checkpoint signaling to ensure accurate chromosome segregation. The kinetochore scaffold Knl1, when phosphorylated by Mps1, recruits checkpoint complexes Bub1–Bub3 and BubR1–Bub3 to unattached kinetochores. Active checkpoint signaling ultimately enhances the assembly of the mitotic checkpoint complex (MCC) consisting of BubR1–Bub3, Mad2, and Cdc20, which inhibits the anaphase-promoting complex or cyclosome bound to Cdc20 (APC/CCdc20) to delay anaphase onset. Using in vitro reconstitution, we show that Mps1 promotes APC/C inhibition by MCC components through phosphorylating Bub1 and Mad1. Phosphorylated Bub1 binds to Mad1–Mad2. Phosphorylated Mad1 directly interacts with Cdc20. Mutations of Mps1 phosphorylation sites in Bub1 or Mad1 abrogate the spindle checkpoint in human cells. Therefore, Mps1 promotes checkpoint activation through sequentially phosphorylating Knl1, Bub1, and Mad1. This sequential multi-target phosphorylation cascade makes the checkpoint highly responsive to Mps1 and to kinetochore-microtubule attachment. DOI: http://dx.doi.org/10.7554/eLife.22513.001 PMID:28072388

Characterization of spindle checkpoint kinase Mps1 reveals domain with functional and structural similarities to tetratricopeptide repeat motifs of Bub1 and BubR1 checkpoint kinases.

PubMed

Lee, Semin; Thebault, Philippe; Freschi, Luca; Beaufils, Sylvie; Blundell, Tom L; Landry, Christian R; Bolanos-Garcia, Victor M; Elowe, Sabine

2012-02-17

Kinetochore targeting of the mitotic kinases Bub1, BubR1, and Mps1 has been implicated in efficient execution of their functions in the spindle checkpoint, the self-monitoring system of the eukaryotic cell cycle that ensures chromosome segregation occurs with high fidelity. In all three kinases, kinetochore docking is mediated by the N-terminal region of the protein. Deletions within this region result in checkpoint failure and chromosome segregation defects. Here, we use an interdisciplinary approach that includes biophysical, biochemical, cell biological, and bioinformatics methods to study the N-terminal region of human Mps1. We report the identification of a tandem repeat of the tetratricopeptide repeat (TPR) motif in the N-terminal kinetochore binding region of Mps1, with close homology to the tandem TPR motif of Bub1 and BubR1. Phylogenetic analysis indicates that TPR Mps1 was acquired after the split between deutorostomes and protostomes, as it is distinguishable in chordates and echinoderms. Overexpression of TPR Mps1 resulted in decreased efficiency of both chromosome alignment and mitotic arrest, likely through displacement of endogenous Mps1 from the kinetochore and decreased Mps1 catalytic activity. Taken together, our multidisciplinary strategy provides new insights into the evolution, structural organization, and function of Mps1 N-terminal region.

Structural and functional insights into the role of the N-terminal Mps1 TPR domain in the SAC (spindle assembly checkpoint).

PubMed

Thebault, Philippe; Chirgadze, Dimitri Y; Dou, Zhen; Blundell, Tom L; Elowe, Sabine; Bolanos-Garcia, Victor M

2012-12-15

The SAC (spindle assembly checkpoint) is a surveillance system that ensures the timely and accurate transmission of the genetic material to offspring. The process implies kinetochore targeting of the mitotic kinases Bub1 (budding uninhibited by benzamidine 1), BubR1 (Bub1 related) and Mps1 (monopolar spindle 1), which is mediated by the N-terminus of each kinase. In the present study we report the 1.8 Å (1 Å=0.1 nm) crystal structure of the TPR (tetratricopeptide repeat) domain in the N-terminal region of human Mps1. The structure reveals an overall high similarity to the TPR motif of the mitotic checkpoint kinases Bub1 and BubR1, and a number of unique features that include the absence of the binding site for the kinetochore structural component KNL1 (kinetochore-null 1; blinkin), and determinants of dimerization. Moreover, we show that a stretch of amino acids at the very N-terminus of Mps1 is required for dimer formation, and that interfering with dimerization results in mislocalization and misregulation of kinase activity. The results of the present study provide an important insight into the molecular details of the mitotic functions of Mps1 including features that dictate substrate selectivity and kinetochore docking.

Meikin-associated polo-like kinase specifies Bub1 distribution in meiosis I.

PubMed

Miyazaki, Seira; Kim, Jihye; Yamagishi, Yuya; Ishiguro, Tadashi; Okada, Yuki; Tanno, Yuji; Sakuno, Takeshi; Watanabe, Yoshinori

2017-06-01

In meiosis I, sister chromatids are captured by microtubules emanating from the same pole (mono-orientation), and centromeric cohesion is protected throughout anaphase. Shugoshin, which is localized to centromeres depending on the phosphorylation of histone H2A by Bub1 kinase, plays a central role in protecting meiotic cohesin Rec8 from separase cleavage. Another key meiotic kinetochore factor, meikin, may regulate cohesion protection, although the underlying molecular mechanisms remain elusive. Here, we show that fission yeast Moa1 (meikin), which associates stably with CENP-C during meiosis I, recruits Plo1 (polo-like kinase) to the kinetochores and phosphorylates Spc7 (KNL1) to accumulate Bub1. Consequently, in contrast to the transient kinetochore localization of mitotic Bub1, meiotic Bub1 persists at kinetochores until anaphase I. The meiotic Bub1 pool ensures robust Sgo1 (shugoshin) localization and cohesion protection at centromeres by cooperating with heterochromatin protein Swi6, which binds and stabilizes Sgo1. Furthermore, molecular genetic analyses show a hierarchical regulation of centromeric cohesion protection by meikin and shugoshin that is important for establishing meiosis-specific chromosome segregation. We provide evidence that the meiosis-specific Bub1 regulation is conserved in mouse. © 2017 Molecular Biology Society of Japan and John Wiley & Sons Australia, Ltd.

Prehepatocholedochal proper hepatic artery. Rare anatomical variant. Surgical considerations. Case report.

PubMed

Ardeleanu, V; Chicoş, S; Tutunaru, D; Georgescu, C

2014-01-01

In classical anatomic variants, the proper hepatic artery (PHA)continues the common hepatic artery (CHA) after the gastroduodenal artery (GDA) detaches itself and divides into the right hepatic artery (RHA) and left hepatic artery (LHA), the proper hepatic artery being located to the left of the hepatocholedochal duct (HCD). This paper presents an abnormal positioning of the PHA placed before the HCD with an increased diameter of about 5-7 mm, which could be confused with the HCD. We present the case of a 57 year-old woman diagnosed with acute lithiasic cholecystitis, associated with hypersplenism and hypertension. The literature mentions manifold anatomical variants of arterial liver vascularization,including PHA. For this reason, this paper presents an overview of similar cases that can be found in medical literature. The aforementioned case is a rare topographic anatomy for the PHA that can easily pass for HCD especially during celioscopy, therefore it is crucial for this to be acknowledged by all surgeons. Celsius.

Access to properly fitting personal protective equipment for female construction workers.

PubMed

Onyebeke, Lynn C; Papazaharias, Demetrios M; Freund, Alice; Dropkin, Jonathan; McCann, Michael; Sanchez, Sadie H; Hashim, Dana; Meyer, John D; Lucchini, Roberto G; Zuckerman, Norman C

2016-11-01

Previous literature suggests that most personal protective equipment (PPE) for construction is designed for males and does not accommodate female anthropometry. We conducted a pilot study to identify whether female construction workers currently have adequate access to properly fitting PPE. Semi-structured focus group interviews were conducted with union female carpenters, laborers, and ironworkers. Researchers coded focus group transcriptions and extracted major themes using thematic framework analysis. Participants (n = 23) had a mean of 15.1 years of construction experience (range 3-34.5 years). A majority reported fit problems for many types of PPE (gloves, harnesses, safety vests, work boots, outerwear), generally noting that the equipment provided by contractors was too large. Other emergent themes included female workers purchasing their own PPE, exposure to various safety hazards from poorly fitted PPE, and perceived indifferent safety culture. Female construction workers continue to have difficulty accessing properly fitting PPE. Am. J. Ind. Med. 59:1032-1040, 2016. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

Guidelines for proper care and use of wildlife in field research

USGS Publications Warehouse

Friend, M.; Toweill, D.E.; Borwnell, R.L.; Nettles, V.F.; Davis, D.S.; Foreyt, W.J.

1999-01-01

Public attitudes towards animals continue to change over time. These changes apply to wildlife along with other species, and in recent years, attitudes have been increasingly oriented toward assuring that all species receive proper care whenever human interactions are involved. Guidance regarding the application of euthanasia is provided in the previous chapter. This chapter provides basic guidelines for the proper use of wildlife in field investigations. We believe this previously published information from The Wildlife Society is sufficiently important to include in this field manual. The Wildlife Society has been kind enough to grant permission for this reproduction. The scope of this chapter extends to all wildlife, and the application of this material extends beyond research to all wildlife investigations. This chapter is reproduced, with the addition of illustrations and minor modifications, as it appeared in Research and Management Techniques for Wildlife and Habitats (Bookhout, 1994), and, thus, it deviates from the format for the rest of Volume I.

Perspectives on barriers and facilitators to self-care in Lebanese cardiac patients: A qualitative descriptive study.

PubMed

Dumit, Nuhad Yazbik; Noureddine, Samar Nayef; Magilvy, Joan Kathy

2016-08-01

Cardiovascular disease is the leading cause of mortality worldwide. Cardiac self-care practices are essential for managing cardiac illness and improving quality of life. However, these practices may be affected by factors that may hinder or facilitate self-care especially in countries that experience political and economic instabilities. The purpose of this study was to explore self-care practices among Lebanese cardiac patients. Another aim was to reveal factors that might influence these self-care practices. This is a qualitative descriptive study. Participants were recruited from a referral medical center in Beirut, Lebanon and interviews took place in their homes. Purposive sample of 15 adult participants, seven females and eight males, diagnosed with coronary artery disease at least a year ago and not in critical condition recruited from the cardiology clinics of the medical center. Data were collected through semi-structured audio-recorded interviews that took place in their places of residents. Three themes emerged from the data: I. The behaviors of cardiac patients demonstrated selected self-care practices; II. Patients identified barriers to self-care reflective of the Lebanese political and socio-economic situation; and, III. Patients described facilitators to self-care consistent with the Lebanese socio-cultural values and norms. The most common self-care practices included taking medications and eating properly. Participants emphasized avoiding stress and being upset as a self-protective measure for cardiac health. Health care costs, family responsibilities, psychological factors and the country's political situation impeded self-care practices whereas family support facilitated them. Lebanese patients reported select self-care practices in dealing with their cardiac illness. Barriers and facilitators to their self-care behaviors reflected the Lebanese context and culture. Thus health care providers must assess their patients' practices within their

Chromosome and mitotic spindle dynamics in fission yeast kinesin-8 mutants

NASA Astrophysics Data System (ADS)

Crapo, Ammon M.; Gergley, Zachary R.; McIntosh, J. Richard; Betterton, M. D.

2014-03-01

Fission yeast proteins Klp5p and Klp6p are plus-end directed motors of the kinesin-8 family which promote microtubule (MT) depolymerization and also affect chromosome segregation, but the mechanism of these activities is not well understood. Using live-cell time-lapse fluorescence microscopy of fission yeast wild-type (WT) and klp5/6 mutant strains, we quantify and compare the dynamics of kinetochore motion and mitotic spindle length in 3D. In WT cells, the spindle, once formed, remains a consistent size and chromosomes are correctly organized and segregated. In kinesin-8 mutants, spindles undergo large length fluctuations of several microns. Kinetochore motions are also highly fluctuating, with kinetochores frequently moving away from the spindle rather than toward it. We observe transient pushing of chromosomes away from the spindle by as much as 10 microns in distance.

42 CFR 405.410 - Conditions for properly opting-out of Medicare.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 42 Public Health 2 2010-10-01 2010-10-01 false Conditions for properly opting-out of Medicare. 405.410 Section 405.410 Public Health CENTERS FOR MEDICARE & MEDICAID SERVICES, DEPARTMENT OF HEALTH AND HUMAN SERVICES MEDICARE PROGRAM FEDERAL HEALTH INSURANCE FOR THE AGED AND DISABLED Private Contracts...

26 CFR 1.1016-2 - Items properly chargeable to capital account.

Code of Federal Regulations, 2010 CFR

2010-04-01

....1016-2 Section 1.1016-2 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES Basis Rules of General Application § 1.1016-2 Items properly... for $10,000. He subsequently expended $6,000 for improvements. Disregarding, for the purpose of this...

Evaluation of the Facilitated Communication Pilot

ERIC Educational Resources Information Center

Cooper-Martin, Elizabeth

2014-01-01

The Office of Special Education and Student Services asked the Office of Shared Accountability to evaluate the "Facilitated Communication Pilot." In facilitated communication (FC), people with communication impairments express themselves by typing with the aid of a communication partner, called a facilitator, who provides physical (and…

Parallaxes and Proper Motions From the MCCormick Observatory: List 47

NASA Astrophysics Data System (ADS)

Ianna, Philip A.; Patterson, Richard J.; Swain, Melanie A.

1996-01-01

Trigonometric parallaxes and proper motions are presented for 32 late-type stars observed photographically with the Leander McCormick 67 cm refractor. Eighteen of the stars have no previously published parallaxes. Twenty one of the stars are K and M dwarfs identified by Vyssotsky and his collaborators. The list includes several x-ray luminous M dwarfs, a rapidly rotating spotted flare star, and two astrometric binaries.

uvby photometry in McCormick proper motion fields

NASA Technical Reports Server (NTRS)

Degewij, J.

1982-01-01

The Danish 50 cm telescope at the European Southern Observatory was used to obtain high-precision uvby photometry for 50 F2 to G2 stars, with V values in the 9.4-12.3 mag range, which were selected in the southern galactic polar regions of the McCormick proper motion fields and measured on six different nights. The brighter stars are found to systematically exhibit smaller m(1) indices, of about 0.02 mag, upon comparison with the earlier data of Blaauw et al (1976). Single measurements are given for 98 stars in eight McCormick fields at intermediate southern galactic latitudes.

Chemically Dissected Rotation Curves of the Galactic Bulge from Main-sequence Proper Motions

NASA Astrophysics Data System (ADS)

Clarkson, William I.; Calamida, Annalisa; Sahu, Kailash C.; Brown, Thomas M.; Gennaro, Mario; Avila, Roberto J.; Valenti, Jeff; Debattista, Victor P.; Rich, R. Michael; Minniti, Dante; Zoccali, Manuela; Aufdemberge, Emily R.

2018-05-01

We report results from an exploratory study implementing a new probe of Galactic evolution using archival Hubble Space Telescope imaging observations. Precise proper motions are combined with photometric relative metallicity and temperature indices, to produce the proper-motion rotation curves of the Galactic bulge separately for metal-poor and metal-rich main-sequence samples. This provides a “pencil-beam” complement to large-scale wide-field surveys, which to date have focused on the more traditional bright giant branch tracers. We find strong evidence that the Galactic bulge rotation curves drawn from “metal-rich” and “metal-poor” samples are indeed discrepant. The “metal-rich” sample shows greater rotation amplitude and a steeper gradient against line-of-sight distance, as well as possibly a stronger central concentration along the line of sight. This may represent a new detection of differing orbital anisotropy between metal-rich and metal-poor bulge objects. We also investigate selection effects that would be implied for the longitudinal proper-motion cut often used to isolate a “pure-bulge” sample. Extensive investigation of synthetic stellar populations suggests that instrumental and observational artifacts are unlikely to account for the observed rotation curve differences. Thus, proper-motion-based rotation curves can be used to probe chemodynamical correlations for main-sequence tracer stars, which are orders of magnitude more numerous in the Galactic bulge than the bright giant branch tracers. We discuss briefly the prospect of using this new tool to constrain detailed models of Galactic formation and evolution. Based on observations made with the NASA/ESA Hubble Space Telescope and obtained from the data archive at the Space Telescope Science Institute. STScI is operated by the Association of Universities for Research in Astronomy, Inc., under NASA contract NAS 5-26555.

[The Pharmaceuticals and Medical Devices Agency's Approach to Facilitate Risk Communication and Its Challenges].

PubMed

Kondo, Emiko; Torii, Mayumi; Oba, Izumi; Okamoto, Mai

2018-01-01

　The issue of drug lag in Japan has been rapidly reduced in recent years, and newly approved drugs now become available on Japanese and international markets at the same time. In this context, the risk management plan (RMP) system was introduced in 2012. RMPs describe important safety concerns recognized by Japan's Pharmaceuticals and Medical Devices Agency (PMDA) and marketing authorization holders (MAHs), as well as safety measures that MAHs request healthcare professionals (HCPs) to follow. The publication of RMPs is expected to support the sharing of drug risk management among HCPs during the postmarketing phase. In addition, to encourage risk communication between HCPs and patients, the PMDA website provides drug guides for patients and other information to promote proper understanding of drugs by patients and their families and enable them to identify serious adverse drug reactions at an early stage. However, the results of surveys conducted by the PMDA in FY2014 and FY2015 revealed low levels of awareness of RMPs and drug guides for patients in hospitals and other healthcare institutions. The surveys also showed that information regarding the proper use of drugs from MAHs and the PMDA was not incorporated into practice at healthcare institutions, resulting in the repeated release of identical safety alerts. To facilitate the increased utilization of risk communication tools, the PMDA has been providing and disseminating these tools through its website. This study addresses those efforts and the associated challenges.

VizieR Online Data Catalog: OGLE high proper motion stars towards MC (Soszynski+, 2002)

NASA Astrophysics Data System (ADS)

Soszynski, I.; Zebrun, K.; Udalski, A.; Wozniak, P. R.; Szymanski, M.; Kubiak, M.; Pietrzynski, G.; Szewczyk, O.; Wyrzykowski, L.

2002-11-01

We present a catalog of high proper motion (HPM) stars detected in the foreground of central parts of the Magellanic Clouds. The Catalog contains 2161 objects in the 4.5 square degree area towards the LMC, and 892 HPM stars in the 2.4 square degree area towards the SMC. The Catalog is based on observations collected during four years of the OGLE-II microlensing survey. The Difference Image Analysis (DIA) of the images provided candidate HPM stars with proper motion as small as 4mas/yr. These appeared as pseudo-variables, and were all measured astrometrically on all CCD images, providing typically about 400 data points per star. The reference frame was defined by the majority of background stars, most of them members of the Magellanic Clouds. The reflex motion due to solar velocity with respect to the local standards of rest is clearly seen. The largest proper motion in our sample is 363mas/yr. Parallaxes were measured with errors smaller than 20% for several stars. (2 data files).

Proper motions and membership probabilities of stars in the region of globular cluster NGC 6366

NASA Astrophysics Data System (ADS)

Sariya, Devesh P.; Yadav, R. K. S.

2015-12-01

Context. NGC 6366 is a metal-rich globular cluster that is relatively unstudied. It is a kinematically interesting cluster, reported as belonging to the slowly rotating halo system, which is unusual given its metallicity and spatial location in the Galaxy. Aims: The purpose of this research is to determine the relative proper motion and membership probability of the stars in the region of globular cluster NGC 6366. To target cluster members reliably during spectroscopic surveys without including field stars, a good proper motion and membership probability catalogue of NGC 6366 is needed. Methods: To derive relative proper motions, the archival data from the Wide Field Imager mounted on the ESO 2.2 m telescope have been reduced using a high precision astrometric software. The images used are in the B,V, and I photometric bands with an epoch gap of ~3.2 yr. The calibrated BVI magnitudes have been determined using recent data for secondary standard stars. Results: We determined relative proper motions and cluster membership probabilities for 2530 stars in the field of globular cluster NGC 6366. The median proper motion rms errors for stars brighter than V ~ 18 mag is ~2 mas yr-1, which gradually increases to ~5 mas yr-1 for stars having magnitudes V ~ 20 mag. Based on the membership catalogue, we checked the membership status of the X-ray sources and variable stars of NGC 6366 mentioned in the literature. We also provide the astronomical community with an electronic catalogue that includes B, V, and I magnitudes; relative proper motions; and membership probabilities of the stars in the region of NGC 6366. Based on observations with the MPG/ESO 2.2 m and ESO/VLT telescopes, located at La Silla and Paranal Observatory, Chile, under DDT programs 164.O-0561(F), 71.D-0220(A) and the archive material.Full Table 4 is only available at the CDS via anonymous ftp to http://cdsarc.u-strasbg.fr (ftp://130.79.128.5) or via http://cdsarc.u-strasbg.fr/viz-bin/qcat?J/A+A/584/A59

UCAC1: New Proper Motions for 27 Million Stars on the Southern Hemisphere

NASA Astrophysics Data System (ADS)

Zacharias, N.; Monet, S. Urban D. G.; Platais, I.; Wycoff, G. L.; Zacharias, M. I.; Rafferty, T. J.

The big impact of UCAC on galactic kinematics and dynamics studies will be outlined. The USNO CCD Astrograph (UCA) started an astrometric sky survey in February 1998 at Cerro Tololo, Chile. By January 2000 about 90% of the Southern Hemisphere has been observed and full sky coverage is expected by early 2003. In addition, calibration fields around extragalactic reference frame sources and selected open clusters are observed frequently. The UCAC project is a huge dedicated astrometric survey similar to the AGK2 and AGK3 projects but vastly exceeding those with respect to higher accuracy, limiting magnitude (16th) and full sky coverage. A first catalog (UCAC1) is being published in early 2000 for 27 million stars. Stars in the range of 9 to 14th magnitude have a positional precision of 20 mas. The UCAC1 will utilize positions from the USNO A2.0 catalog for determining proper motions, which are expected to be about 8 mas/yr for this initial release. Higher precision proper motions, expected to be in the 3 to 4 mas/yr range, will be derived utilizing a variety of early epoch data, including re-measuring of the Southern Proper Motion (SPM) survey first epoch plates.

VizieR Online Data Catalog: New proper motion stars with pm>=0.18"/yr (Boyd+, 2011)

NASA Astrophysics Data System (ADS)

Boyd, M. R.; Henry, T. J.; Jao, W.-C.; Subasavage, J. P.; Hambly, N. C.

2012-11-01

Here we present 1584 new southern proper motion systems with μ>=0.18"/yr and 16.5>R59F>=18.0. This search complements the six previous SuperCOSMOS-RECONS (SCR) proper motion searches of the southern sky for stars within the same proper motion range, but with R59F<=16.5. As in previous papers, we present distance estimates for these systems and find that three systems are estimated to be within 25pc, including one, SCR 1546-5534, possibly within the RECONS 10pc horizon at 6.7pc, making it the second nearest discovery of the searches. We find 97 white dwarf candidates with distance estimates between 10 and 120pc, as well as 557 cool subdwarf candidates. (5 data files).

Group Facilitation: Functions and Skills.

ERIC Educational Resources Information Center

Anderson, L. Frances; Robertson, Sharon E.

1985-01-01

Discusses a model based on a specific set of assumptions about causality and effectiveness in interactional groups. Discusses personal qualities of group facilitators and proposes five major functions and seven skill clusters central to effective group facilitation. (Author/BH)

Effects of Student-Facilitated Learning on Instructional Facilitators

ERIC Educational Resources Information Center

Bonner, Sarah M.; Somers, Jennifer A.; Rivera, Gwendelyn J.; Keiler, Leslie S.

2017-01-01

We investigated perceptions about learning strategy use and instructional roles among a sample of high needs adolescents (n = 230) who acted as near-peer instructional facilitators. The sample was drawn from science and mathematics classes in nonselective public secondary schools in New York City. Students participated in an inschool intervention…

Barriers to and facilitators of HIV-positive patients' adherence to antiretroviral treatment regimens.

PubMed

Roberts, K J

2000-03-01

HIV-positive patients must strictly adhere to antiretroviral regimens for the medications to work properly. Little, however, is known about the obstacles that patients face in adhering to the regimens or what, if anything, helps patients to adhere. The goals of the project were to describe, from HIV-positive patients' own perspectives, the barriers they face in adhering to antiretroviral regimens and the strategies they use to maximize their adherence. Five main barriers (forgetfulness, social/physical environment, complexity of the regimens, medication side effects, and inadequate patient knowledge) to adherence and six main facilitators (mechanical devices, "making a commitment," "routinizing," health beliefs, social support, and professional support) emerged from the data. Patients may overcome some of these barriers by receiving better health education about the need for adherence, professional and lay support for their efforts, and mechanical devices such as alarm clocks and medi-sets. Other barriers, however, such as the complexity of the medications, highlight the need for simplified antiretroviral regimens.

The c-index is not proper for the evaluation of $t$-year predicted risks.

PubMed

Blanche, Paul; Kattan, Michael W; Gerds, Thomas A

2018-02-16

We show that the widely used concordance index for time to event outcome is not proper when interest is in predicting a $t$-year risk of an event, for example 10-year mortality. In the situation with a fixed prediction horizon, the concordance index can be higher for a misspecified model than for a correctly specified model. Impropriety happens because the concordance index assesses the order of the event times and not the order of the event status at the prediction horizon. The time-dependent area under the receiver operating characteristic curve does not have this problem and is proper in this context.

Photometric detection of high proper motions in dense stellar fields using difference image analysis

NASA Astrophysics Data System (ADS)

Eyer, L.; Woźniak, P. R.

2001-10-01

The difference image analysis (DIA) of the images obtained by the Optical Gravitational Lensing Experiment (OGLE-II) revealed a peculiar artefact in the sample of stars proposed as variable by Woźniak in one of the Galactic bulge fields: the occurrence of pairs of candidate variables showing anti-correlated light curves monotonic over a period of 3yr. This effect can be understood, quantified and related to the stellar proper motions. DIA photometry supplemented with a simple model offers an effective and easy way to detect high proper motion stars in very dense stellar fields, where conventional astrometric searches are extremely inefficient.

Artificial ascites and pneumoperitoneum to facilitate thermal ablation of liver tumors: a pictorial essay.

PubMed

Bhagavatula, Sharath K; Chick, Jeffrey F B; Chauhan, Nikunj R; Shyn, Paul B

2017-02-01

Image-guided percutaneous thermal ablation is increasingly utilized in the treatment of hepatic malignancies. Peripherally located hepatic tumors can be difficult to access or located adjacent to critical structures that can be injured. As a result, ablation of peripheral tumors may be avoided or may be performed too cautiously, leading to inadequate ablation coverage. In these cases, separating the tumor from adjacent critical structures can increase the efficacy and safety of procedures. Artificial ascites and artificial pneumoperitoneum are techniques that utilize fluid and gas, respectively, to insulate critical structures from the thermal ablation zone. Induction of artificial ascites and artificial pneumoperitoneum can enable complete ablation of otherwise inaccessible hepatic tumors, improve tumor visualization, minimize unintended thermal injury to surrounding organs, and reduce post-procedural pain. This pictorial essay illustrates and discusses the proper technique and clinical considerations for successful artificial ascites and pneumoperitoneum creation to facilitate safe peripheral hepatic tumor ablation.

Regulators of spindle microtubules and their mechanisms: Living together matters.

PubMed

Lakshmi, R Bhagya; Nair, Vishnu M; Manna, Tapas K

2018-02-01

Development and survival of all eukaryotic organisms depend on equal partitioning of their chromosomes between the two newly formed daughter cells during mitosis. The mitotic spindle performs the task of physically segregating the chromosomes through multiple stages of mitosis. During this process, kinetochore-microtubule attachment requires to be selectively stabilized to hold the chromosomes, but at the same time, it has to be flexible enough to allow kinetochore microtubule dynamicity and chromosome movements. Research during the last decade or so has identified a number of proteins associated with the spindle microtubule plus ends that regulate these processes and orchestrate forces to spatially organize and separate the chromosomes. In this review, we describe the molecular details of those regulators and their mechanisms of action at the kinetochore-microtubule interface. © 2018 IUBMB Life, 70(2):101-111, 2018. © 2018 International Union of Biochemistry and Molecular Biology.

Drosophila parthenogenesis: A tool to decipher centrosomal vs acentrosomal spindle assembly pathways

DOE Office of Scientific and Technical Information (OSTI.GOV)

Riparbelli, Maria Giovanna; Callaini, Giuliano

2008-04-15

Development of unfertilized eggs in the parthenogenetic strain K23-O-im of Drosophila mercatorum requires the stochastic interactions of self-assembled centrosomes with the female chromatin. In a portion of the unfertilized eggs that do not assemble centrosomes, microtubules organize a bipolar anastral mitotic spindle around the chromatin like the one formed during the first female meiosis, suggesting that similar pathways may be operative. In the cytoplasm of eggs in which centrosomes do form, monastral and biastral spindles are found. Analysis by laser scanning confocal microscopy suggests that these spindles are derived from the stochastic interaction of astral microtubules directly with kinetochore regionsmore » or indirectly with kinetochore microtubules. Our findings are consistent with the idea that mitotic spindle assembly requires both acentrosomal and centrosomal pathways, strengthening the hypothesis that astral microtubules can dictate the organization of the spindle by capturing kinetochore microtubules.« less

Holocentromere identity: from the typical mitotic linear structure to the great plasticity of meiotic holocentromeres.

PubMed

Marques, André; Pedrosa-Harand, Andrea

2016-09-01

The centromere is the chromosomal site of kinetochore assembly and is responsible for the correct chromosome segregation during mitosis and meiosis in eukaryotes. Contrary to monocentrics, holocentric chromosomes lack a primary constriction, what is attributed to a kinetochore activity along almost the entire chromosome length during mitosis. This extended centromere structure imposes a problem during meiosis, since sister holocentromeres are not co-oriented during first meiotic division. Thus, regardless of the relatively conserved somatic chromosome structure of holocentrics, during meiosis holocentric chromosomes show different adaptations to deal with this condition. Recent findings in holocentrics have brought back the discussion of the great centromere plasticity of eukaryotes, from the typical CENH3-based holocentromeres to CENH3-less holocentric organisms. Here, we summarize recent and former findings about centromere/kinetochore adaptations shown by holocentric organisms during mitosis and meiosis and discuss how these adaptations are related to the type of meiosis found.

Centromeric Heterochromatin: The Primordial Segregation Machine

PubMed Central

Bloom, Kerry S.

2014-01-01

Centromeres are specialized domains of heterochromatin that provide the foundation for the kinetochore. Centromeric heterochromatin is characterized by specific histone modifications, a centromere-specific histone H3 variant (CENP-A), and the enrichment of cohesin, condensin, and topo-isomerase II. Centromere DNA varies orders of magnitude in size from 125 bp (budding yeast) to several megabases (human). In metaphase, sister kinetochores on the surface of replicated chromosomes face away from each other, where they establish microtubule attachment and bi-orientation. Despite the disparity in centromere size, the distance between separated sister kinetochores is remarkably conserved (approximately 1 μm) throughout phylogeny. The centromere functions as a molecular spring that resists microtubule-based extensional forces in mitosis. This review explores the physical properties of DNA in order to understand how the molecular spring is built and how it contributes to the fidelity of chromosome segregation. PMID:25251850

Myrtenal, a controversial molecule for the proper application of the CIP Sequence Rule for multiple bonds.

PubMed

Zepeda, L Gerardo; Burgueño-Tapia, Eleuterio; Joseph-Nathan, Pedro

2011-04-01

This communication highlights the need of building hierarchical digraphs for the unequivocal assignment of stereochemical descriptors of (-)-myrtenal, a naturally-occurring oxygenated monoterpene whose absolute configuration (AC) is sometimes misrepresented in its structural formulae. Differentiation between duplicated atoms and phantom atoms for the proper application of the sequence rules is shown to be an essential step to get a proper construction of hierarchical digraphs.

VizieR Online Data Catalog: Proper motions of PM2000 open clusters (Krone-Martins+, 2010)

NASA Astrophysics Data System (ADS)

Krone-Martins, A.; Soubiran, C.; Ducourant, C.; Teixeira, R.; Le Campion, J. F.

2010-04-01

We present lists of proper-motions and kinematic membership probabilities in the region of 49 open clusters or possible open clusters. The stellar proper motions were taken from the Bordeaux PM2000 catalogue. The segregation between cluster and field stars and the assignment of membership probabilities was accomplished by applying a fully automated method based on parametrisations for the probability distribution functions and genetic algorithm optimisation heuristics associated with a derivative-based hill climbing algorithm for the likelihood optimization. (3 data files).

Messier 35 (NGC 2168) DANCe. I. Membership, proper motions, and multiwavelength photometry

NASA Astrophysics Data System (ADS)

Bouy, H.; Bertin, E.; Barrado, D.; Sarro, L. M.; Olivares, J.; Moraux, E.; Bouvier, J.; Cuillandre, J.-C.; Ribas, Á.; Beletsky, Y.

2015-03-01

Context. Messier 35 (NGC 2168) is an important young nearby cluster. Its age, richness and relative proximity make it an ideal target for stellar evolution studies. The Kepler K2 mission recently observed it and provided a high accuracy photometric time series of a large number of sources in this area of the sky. Identifying the cluster's members is therefore of high importance to optimize the interpretation and analysis of the Kepler K2 data. Aims: We aim to identify the cluster's members by deriving membership probabilities for the sources within 1° of the cluster's center, which is farther away than equivalent previous studies. Methods: We measure accurate proper motions and multiwavelength (optical and near-infrared) photometry using ground-based archival images of the cluster. We use these measurements to compute membership probabilities. The list of candidate members from the literature is used as a training set to identify the cluster's locus in a multidimensional space made of proper motions, luminosities, and colors. Results: The final catalog includes 338 892 sources with multiwavelength photometry. Approximately half (194 452) were detected at more than two epochs and we measured their proper motion and used it to derive membership probability. A total of 4349 candidate members with membership probabilities greater than 50% are found in this sample in the luminosity range between 10 mag and 22 mag. The slow proper motion of the cluster and the overlap of its sequence with the field and background sequences in almost all color-magnitude and color-color diagrams complicate the analysis and the contamination level is expected to be significant. Our study, nevertheless, provides a coherent and quantitative membership analysis of Messier 35 based on a large fraction of the best ground-based data sets obtained over the past 18 years. As such, it represents a valuable input for follow-up studies using, in particular, the Kepler K2 photometric time series

Reduced verbal fluency for proper names in nondemented patients with Parkinson's disease: a quantitative and qualitative analysis.

PubMed

Fine, Eric M; Delis, Dean C; Paul, Brianna M; Filoteo, J Vincent

2011-02-01

There has been an increasing interest within neuropsychology in comparing verbal fluency for different grammatical classes (e.g., verb generation vs. noun generation) in neurological populations, including Parkinson's disease (PD). However, to our knowledge, few studies have compared verbal fluency for common nouns and proper names in PD. Common nouns and proper names differ in terms of their semantic characteristics, as categories of common nouns are organized hierarchically based on semantics, while categories of proper nouns lack a well-defined semantic organization. In addition, there is accumulating evidence that the retrieval of these distinct grammatical classes are subserved by somewhat distinct neural systems. Given that verbal fluency deficits are among the first impairments to emerge in PD, and that such deficits are predictors of future cognitive decline, it is important to examine all aspects of verbal fluency in this population. For the current study, we compared the performance of a group of 32 nondemented PD patients with 32 healthy participants (HP) on verbal fluency tasks for common nouns (animals) and proper names (boys' first names). A significant interaction between verbal fluency task and diagnostic status emerged, as the PD group performed significantly worse on only the proper name fluency task. This finding may reflect the absence of well-defined semantic organization that structures the verbal search for first names, thus placing a greater onus on strategic or "executive" verbal retrieval processes.

Proper muscle layer damage affects ulcer healing after gastric endoscopic submucosal dissection.

PubMed

Horikawa, Yohei; Mimori, Nobuya; Mizutamari, Hiroya; Kato, Yuhei; Shimazu, Kazuhiro; Sawaguchi, Masayuki; Tawaraya, Shin; Igarashi, Kimihiro; Okubo, Syunji

2015-11-01

Endoscopic submucosal dissection (ESD) is the established therapy for superficial gastrointestinal neoplasms. However, management of the artificial ulcers associated with ESD has become important and the relationship between ulcer healing factors and treatment is still unclear. We aimed to evaluate ESD-related artificial ulcer reduction ratio at 4 weeks to assess factors associating with ulcer healing after ESD that may lead to optimal treatment. Between January 2009 and December 2013, a total of 375 lesions fulfilled the expanded criteria for ESD. We defined ulcer reduction rate <90% as (A) poor-healing group; and rate ≥90% as (B) well-healing group. After exclusion, 328 lesions were divided into two groups and analyzed. These two groups were compared based on clinicopathological/endoscopic features, concomitant drugs, and treatment. Ulcer reduction rate was significantly correlated with factors related to the ESD procedure (i.e. procedure time, submucosal fibrosis, and injury of the proper muscle layer, in univariate analysis. Multivariate logistic regression analysis showed that submucosal fibrosis (F2) (P = 0.03; OR, 16.46; 95% CI, 1.31-206.73) and injury of the proper muscle layer (P = 0.01; OR, 4.27; 95% CI, 2.04-8.92) were statistically significant predictors of delayed healing. This single-center retrospective study indicated that ESD-induced artificial ulcer healing was affected by submucosal fibrosis and injury of the proper muscle layer, which induced damage to the muscle layer. Therefore, the preferable pharmacotherapy can be determined on completion of the ESD procedure. © 2015 The Authors Digestive Endoscopy © 2015 Japan Gastroenterological Endoscopy Society.

Facilitated Communication: An Experimental Evaluation.

ERIC Educational Resources Information Center

Regal, Robert A.; And Others

1994-01-01

Nineteen adults with developmental disabilities, judged competent in facilitated communication, participated in a validation study using an information passing design requiring short-term recall of stimulus cards with shapes, colors, and numbers. Results failed to validate facilitated communication for the group as a whole, any individual…

Virtual OD: Facilitating Groups Online

ERIC Educational Resources Information Center

Milton, Judy; Watkins, Karen E.; Daley, Barbara J.

2005-01-01

This study examined the role of facilitators in nine virtual action learning groups. A qualitative analysis of the facilitators' interventions across all groups resulted in a typology that included group management, group process, and support interventions. A model showing the relationship among these categories proposes that effective…

The Liverpool-Edinburgh high proper motion catalogue

NASA Astrophysics Data System (ADS)

Pokorny, R. S.; Jones, H. R. A.; Hambly, N. C.; Pinfield, D. J.

2004-07-01

We present a machine selected catalogue of 11 289 objects with proper motions exceeding 0.18 arcsec yr-1 and an R-band faint magnitude limit of 19.5 mag. The catalogue was produced using SuperCOSMOS digitized R-Band ESO and UK Schmidt Plates in 287 Schmidt fields covering almost 7000 square degrees (˜17% of the whole sky) at the South Galactic Cap. The catalogue includes UK Schmidt BJ and I magnitudes for all of the stars as well as 2MASS magnitudes for 10,447 of the catalogue stars. We also show that the NLTT is ˜95% complete for Dec > -32.5°. The full Table \\ref{tab1} is only available in electronic form at the CDS via anonymous ftp to cdsarc.u-strasbg.fr (130.79.128.5) or via http://cdsweb.u-strasbg.fr/cgi-bin/qcat?J/A+A/421/763

Colchicine promotes a change in chromosome structure without loss of sister chromatid cohesion in prometaphase I-arrested bivalents.

PubMed

Rodríguez, E M; Parra, M T; Rufas, J S; Suja, J A

2001-12-01

In somatic cells colchicine promotes the arrest of cell division at prometaphase, and chromosomes show a sequential loss of sister chromatid arm and centromere cohesion. In this study we used colchicine to analyse possible changes in chromosome structure and sister chromatid cohesion in prometaphase I-arrested bivalents of the katydid Pycnogaster cucullata. After silver staining we observed that in colchicine-arrested prometaphase I bivalents, and in contrast to what was found in control bivalents, sister kinetochores appeared individualised and sister chromatid axes were completely separated all along their length. However, this change in chromosome structure occurred without loss of sister chromatid arm cohesion. We also employed the MPM-2 monoclonal antibody against mitotic phosphoproteins on control and colchicine-treated spermatocytes. In control metaphase I bivalents this antibody labelled the tightly associated sister kinetochores and the interchromatid domain. By contrast, in colchicine-treated prometaphase I bivalents individualised sister kinetochores appeared labelled, but the interchromatid domain did not show labelling. These results support the notion that MPM-2 phosphoproteins, probably DNA topoisomerase IIalpha, located in the interchromatid domain act as "chromosomal staples" associating sister chromatid axes in metaphase I bivalents. The disappearance of these chromosomal staples would induce a change in chromosome structure, as reflected by the separation of sister kinetochores and sister axes, but without a concomitant loss of sister chromatid cohesion.

X-ray Proper Motions and Shock Speeds along the Northwest Rim of SN1006

NASA Technical Reports Server (NTRS)

Katsuda, Satoru; Long, Knox S.; Petre, Robert; Reynolds, Stephen P.; Williams, Brian J.; Winkler, P. Frank

2012-01-01

We report the results of an X-ray proper motion measurement for the NW rim of SN 1006, carried out by comparing Chandra observations from 2001 and 2012. The NW limb has predominantly thermal X-ray emission, and it is the only location in SN 1006 with signi cant optical emission: a thin, Balmer-dominated lament. For most of the NW rim, the proper motion is approximately equal to 0.30"yr(exp -1), essentially the same as has been measured from the H alpha lament. Isolated regions of the NW limb are dominated by nonthermal emission, and here the proper motion is much higher, 0:49"yr(exp -1), close to the value measured in X-rays along the much brighter NE limb, where the X-rays are overwhelmingly nonthermal. At the 2.2 kpc distance to SN 1006, the proper motions imply shock velocities of approximately 3000 kms(exp -1) and approximately 5000 kms(exp -1) in the thermal and nonthermal regions, respectively. A lower velocity behind the H alpha filament is consistent with the picture that SN 1006 is encountering denser gas in the NW, as is also suggested by its overall morphology. In the thermally-dominated portion of the X-ray shell, we also see an o set in the radial profiles at different energies; the 0.5-0.6 keV peak dominated by O VII is closer to the shock front than that of the 0.8-3 keV emission|due to the longer times for heavier elements to reach ionization states where they produce strong X-ray emission.

Facilitating Dialogues about Racial Realities

ERIC Educational Resources Information Center

Quaye, Stephen John

2014-01-01

Background/Context: Facilitating dialogues about racial issues in higher education classroom settings continues to be a vexing problem facing postsecondary educators. In order for students to discuss race with their peers, they need skilled facilitators who are knowledgeable about racial issues and able to support students in these difficult…

The Essential Elements of Facilitation.

ERIC Educational Resources Information Center

Priest, Simon; Gass, Michael; Gillis, Lee

Most organizations find it difficult to implement change, and only about 10 percent of learning from training and development experiences is actually applied in the workplace. This book advocates facilitation as a means of enhancing change and increasing productivity. Facilitation engages employees by enhancing the processes associated with their…

Mps1 promotes rapid centromere accumulation of Aurora B.

PubMed

van der Waal, Maike S; Saurin, Adrian T; Vromans, Martijn J M; Vleugel, Mathijs; Wurzenberger, Claudia; Gerlich, Daniel W; Medema, René H; Kops, Geert J P L; Lens, Susanne M A

2012-09-01

Aurora B localization to mitotic centromeres, which is required for proper chromosome alignment during mitosis, relies on Haspin-dependent histone H3 phosphorylation and on Bub1-dependent histone H2A phosphorylation--which interacts with Borealin through a Shugoshin (Sgo) intermediate. We demonstrate that Mps1 stimulates the latter recruitment axis. Mps1 activity enhances H2A-T120ph and is critical for Sgo1 recruitment to centromeres, thereby promoting Aurora B centromere recruitment in early mitosis. Importantly, chromosome biorientation defects caused by Mps1 inhibition are improved by restoring Aurora B centromere recruitment. As Mps1 kinetochore localization reciprocally depends on Aurora B, we propose that this Aurora B-Mps1 recruitment circuitry cooperates with the Aurora B-Haspin feedback loop to ensure rapid centromere accumulation of Aurora B at the onset of mitosis.

Proper Motions of Dwarf Spheroidal Galaxies from Hubble Space Telescope Imaging. II. Measurement for Carina

NASA Astrophysics Data System (ADS)

Piatek, Slawomir; Pryor, Carlton; Olszewski, Edward W.; Harris, Hugh C.; Mateo, Mario; Minniti, Dante; Tinney, Christopher G.

2003-11-01

This article presents and discusses a measurement of the proper motion for the Carina dwarf spheroidal galaxy (dSph) from images in two distinct fields in the direction of Carina taken with the Hubble Space Telescope, at three epochs. Each field contains a confirmed quasi-stellar object that is the reference point for measuring the proper motion of the dSph. The consecutive epochs are 1-2 yr apart. The components of the measured proper motion for Carina, expressed in the equatorial coordinate system, are μα=22+/-9 mas century-1 and μδ=15+/-9 mas century-1. The quoted proper motion is a weighted mean of two independent measurements and has not been corrected for the motions of the Sun and of the local standard of rest. Given the proper motion and its uncertainty, integrating the family of possible orbits of Carina in a realistic gravitational potential for the Milky Way indicates that Carina is bound gravitationally to the Milky Way and is close to apogalacticon. The best estimate of, and the 95% confidence interval for, the apogalacticon of the orbit is 102 kpc and (102,113) kpc, for the perigalacticon is 20 kpc and (3.0,63) kpc, and for the orbital period is 1.4 Gyr and (1.3,2.0) Gyr. Carina does not seem to be on a polar orbit. The best estimate of the inclination of the orbit with respect to the Galactic plane is 39°, but the 95% confidence interval is so wide, (23°,102°), that it includes a polar orbit. We are unable to confirm or to rule out the membership of Carina in a ``stream'' of galaxies in the Galactic halo because the difference between the observed and predicted directions of the proper motion is 1.6 times the uncertainty of the difference. Carina must contain dark matter to have survived the tidal interaction with the Milky Way until the present. The triggering of star formation by perigalacticon passages and crossings of the Galactic disk do not explain the history of star formation in Carina. Based on observations with NASA/ESA Hubble Space

Clinical education facilitators: a literature review.

PubMed

Lambert, Veronica; Glacken, Michèle

2005-07-01

The aim of this literature review, set within an Irish context, is to present a broad overview of former and existing clinical support personnel, explore the concept of facilitation and examine what is known about the role of the clinical education facilitator. The importance of providing a supportive clinical environment to enhance clinical teaching and learning is strongly portrayed in the literature. While the past two decades have borne witness to various clinical support personnel, the literature identifies conflicting demands that these personnel face. No suggestions are advanced as to how to overcome these difficulties, which inevitably influence the quality and quantity of their clinical teaching role. An identifiable gap exists over who has prime responsibility for clinical teaching. It is timely that alternative possibilities for organizing clinical teaching are investigated. A new post emerging in practice settings is that of the clinical education facilitator who is meant to be the key linchpin in clinical areas for reducing the theory-practice gap. Relevant literature for this review was sourced using the computerized databases CINAHL, Medline and Synergy. Manual searching of relevant nursing journals and sourcing of secondary references extended the search. Government reports and other relevant documents were obtained through pertinent websites. Papers that explicitly examined the concept of facilitation and explored the posts of clinical education facilitators were included; six research papers were accessed and reviewed. In addition seven non-empirical papers were included. It is clear that considerable lack of role clarity resides over what constitutes clinical facilitation and the role of the clinical facilitator. Thus, it is paramount to strengthen this support role with Irish empirical evidence. A major advantage in having a ward-based clinical education facilitator is the benefit of having access to someone who can concentrate solely on

Proper-Motion Study of the Magellanic Clouds Using SPM Material

DTIC Science & Technology

2010-12-01

F. VAN ALTENA I, NORBERT ZACHARIAS2, DANA I. CASETTI-DINESCU I, VLADIMIR I. KORCHAGIN I, iMANTSPLATAIS3, DAVID G. MONE -r4, CARLOS E. LOPEZ5, DAVID...difference of the two clouds to within ±54 km S-I. The absolute proper-motion results are consistent with the Clouds’ orbits being marginally bound...consistent with the Clouds’ orbits being marginally bound to the Milky Way, albeit on an elongated orbit. The inferred relative velocity between the

Constrained reduced-order models based on proper orthogonal decomposition

DOE PAGES

Reddy, Sohail R.; Freno, Brian Andrew; Cizmas, Paul G. A.; ...

2017-04-09

A novel approach is presented to constrain reduced-order models (ROM) based on proper orthogonal decomposition (POD). The Karush–Kuhn–Tucker (KKT) conditions were applied to the traditional reduced-order model to constrain the solution to user-defined bounds. The constrained reduced-order model (C-ROM) was applied and validated against the analytical solution to the first-order wave equation. C-ROM was also applied to the analysis of fluidized beds. Lastly, it was shown that the ROM and C-ROM produced accurate results and that C-ROM was less sensitive to error propagation through time than the ROM.

Delamination properity of 2060 aluminium lithium alloy laminate

NASA Astrophysics Data System (ADS)

Guo, Xiang; Huang, Yi; Tong, Dihua; Ma, Shaojun

2018-03-01

Delamination is an significant property of aluminium lithium alloy laminate for damage tolerance design. Double Crack Lap Shear specimens of "2/1" structure are used for testing delamination properity. Experiments of 2 stress ratio conditions R=0.06, 0.5 are conducted, and 5 stress levels for each stress ratio. Delamination growth data b-N are obtained from 4 crack tips’ locations. The energy release rate, Gd, of the "2/1" structure laminate are calculated. Alderliesten modelis used for describing delamination growth db/dN and energy release rate Gd. Parameters Cd and nd in Alderliesten model are determined for R=0.06 and R=0.5 by linear fitting. An analysis is made by comparing the results of different stress ratio conditons.

Gravitational Scattering Amplitudes and Closed String Field Theory in the Proper-Time Gauge

NASA Astrophysics Data System (ADS)

Lee, Taejin

2018-01-01

We construct a covariant closed string field theory by extending recent works on the covariant open string field theory in the proper-time gauge. Rewriting the string scattering amplitudes generated by the closed string field theory in terms of the Polyakov string path integrals, we identify the Fock space representations of the closed string vertices. We show that the Fock space representations of the closed string field theory may be completely factorized into those of the open string field theory. It implies that the well known Kawai-Lewellen-Tye (KLT) relations of the first quantized string theory may be promoted to the second quantized closed string theory. We explicitly calculate the scattering amplitudes of three gravitons by using the closed string field theory in the proper-time gauge.

Msc1 acts through histone H2A.Z to promote chromosome stability in Schizosaccharomyces pombe.

PubMed

Ahmed, Shakil; Dul, Barbara; Qiu, Xinxing; Walworth, Nancy C

2007-11-01

As a central component of the DNA damage checkpoint pathway, the conserved protein kinase Chk1 mediates cell cycle progression when DNA damage is generated. Msc1 was identified as a multicopy suppressor capable of facilitating survival in response to DNA damage of cells mutant for chk1. We demonstrate that loss of msc1 function results in an increased rate of chromosome loss and that an msc1 null allele exhibits genetic interactions with mutants in key kinetochore components. Multicopy expression of msc1 robustly suppresses a temperature-sensitive mutant (cnp1-1) in the centromere-specific histone H3 variant CENP-A, and localization of CENP-A to the centromere is compromised in msc1 null cells. We present several lines of evidence to suggest that Msc1 carries out its function through the histone H2A variant H2A.Z, encoded by pht1 in fission yeast. Like an msc1 mutant, a pht1 mutant also exhibits chromosome instability and genetic interactions with kinetochore mutants. Suppression of cnp1-1 by multicopy msc1 requires pht1. Likewise, suppression of the DNA damage sensitivity of a chk1 mutant by multicopy msc1 also requires pht1. We present the first genetic evidence that histone H2A.Z may participate in centromere function in fission yeast and propose that Msc1 acts through H2A.Z to promote chromosome stability and cell survival following DNA damage.

Msc1 Acts Through Histone H2A.Z to Promote Chromosome Stability in Schizosaccharomyces pombe

PubMed Central

Ahmed, Shakil; Dul, Barbara; Qiu, Xinxing; Walworth, Nancy C.

2007-01-01

As a central component of the DNA damage checkpoint pathway, the conserved protein kinase Chk1 mediates cell cycle progression when DNA damage is generated. Msc1 was identified as a multicopy suppressor capable of facilitating survival in response to DNA damage of cells mutant for chk1. We demonstrate that loss of msc1 function results in an increased rate of chromosome loss and that an msc1 null allele exhibits genetic interactions with mutants in key kinetochore components. Multicopy expression of msc1 robustly suppresses a temperature-sensitive mutant (cnp1-1) in the centromere-specific histone H3 variant CENP-A, and localization of CENP-A to the centromere is compromised in msc1 null cells. We present several lines of evidence to suggest that Msc1 carries out its function through the histone H2A variant H2A.Z, encoded by pht1 in fission yeast. Like an msc1 mutant, a pht1 mutant also exhibits chromosome instability and genetic interactions with kinetochore mutants. Suppression of cnp1-1 by multicopy msc1 requires pht1. Likewise, suppression of the DNA damage sensitivity of a chk1 mutant by multicopy msc1 also requires pht1. We present the first genetic evidence that histone H2A.Z may participate in centromere function in fission yeast and propose that Msc1 acts through H2A.Z to promote chromosome stability and cell survival following DNA damage. PMID:17947424

General relativistic satellite astrometry. II. Modeling parallax and proper motion

NASA Astrophysics Data System (ADS)

de Felice, F.; Bucciarelli, B.; Lattanzi, M. G.; Vecchiato, A.

2001-07-01

The non-perturbative general relativistic approach to global astrometry introduced by de Felice et al. (\\cite{defetal}) is here extended to account for the star motions on the Schwarzschild celestial sphere. A new expression of the observables, i.e. angular distances among stars, is provided, which takes into account the effects of parallax and proper motions. This dynamical model is then tested on an end-to-end simulation of the global astrometry mission GAIA. The results confirm the findings of our earlier work, which applied to the case of a static (angular coordinates only) sphere. In particular, measurements of large arcs among stars (each measurement good to ~ 100 mu arcsec, as expected for V ~ 17 mag stars) repeated over an observing period comparable to the mission lifetime foreseen for GAIA, can be modeled to yield estimates of positions, parallaxes, and annual proper motions good to ~ 15 mu arcsec. This second round of experiments confirms, within the limitations of the simulation and the assumptions of the current relativistic model, that the space-born global astrometry initiated with Hipparcos can be pushed down to the 10-5 arcsec accuracy level proposed with the GAIA mission. Finally, the simplified case we have solved can be used as reference for testing the limiting behavior of more realistic models as they become available.

SIMP: A Near-Infrared Proper Motion Survey

NASA Astrophysics Data System (ADS)

Artigau, Étienne; Lafrenière, David; Doyon, René; Albert, Loïc; Robert, Jasmin; Malo, Lison

2009-02-01

SIMP is a proper motion (PM) survey made with the Observatoire du Mont Mégantic (OMM) wide-field near-infrared camera CPAPIR at the CTIO 1.5 m and OMM 1.6 m telescopes. The SIMP observations were initiated in early 2005, are still ongoing and, to date, have covered 28% of the sky at high galactic latitudes. The PMs of the sources detected are determined by comparing their measured positions with those listed in the 2MASS point source catalog, giving a time baseline of 4 to 10 years. The 5 σ uncertainty on the relative SIMP and 2MASS astrometry is 1'', equivalent to a PM lower limit of 0.125-0.250''/yr, or a tangential velocity limit of 15-30 km/s at 25 pc. Up to the 2MASS magnitude limit (J~16.5), T dwarfs are found out to ~25 pc, while L dwarfs may be found as far as 100 pc away.

Direct measurement of conformational strain energy in protofilaments curling outward from disassembling microtubule tips.

PubMed

Driver, Jonathan W; Geyer, Elisabeth A; Bailey, Megan E; Rice, Luke M; Asbury, Charles L

2017-06-19

Disassembling microtubules can generate movement independently of motor enzymes, especially at kinetochores where they drive chromosome motility. A popular explanation is the 'conformational wave' model, in which protofilaments pull on the kinetochore as they curl outward from a disassembling tip. But whether protofilaments can work efficiently via this spring-like mechanism has been unclear. By modifying a previous assay to use recombinant tubulin and feedback-controlled laser trapping, we directly demonstrate the spring-like elasticity of curling protofilaments. Measuring their mechanical work output suggests they carry ~25% of the energy of GTP hydrolysis as bending strain, enabling them to drive movement with efficiency similar to conventional motors. Surprisingly, a β-tubulin mutant that dramatically slows disassembly has no effect on work output, indicating an uncoupling of disassembly speed from protofilament strain. These results show the wave mechanism can make a major contribution to kinetochore motility and establish a direct approach for measuring tubulin mechano-chemistry.

Structure of the RZZ complex and molecular basis of its interaction with Spindly

PubMed Central

Mosalaganti, Shyamal; Keller, Jenny; Altenfeld, Anika; Rombaut, Pascaline; Petrovic, Arsen; Wohlgemuth, Sabine; Müller, Franziska; Herzog, Franz; Waldmann, Herbert

2017-01-01

Kinetochores are macromolecular assemblies that connect chromosomes to spindle microtubules (MTs) during mitosis. The metazoan-specific ≈800-kD ROD–Zwilch–ZW10 (RZZ) complex builds a fibrous corona that assembles on mitotic kinetochores before MT attachment to promote chromosome alignment and robust spindle assembly checkpoint signaling. In this study, we combine biochemical reconstitutions, single-particle electron cryomicroscopy, cross-linking mass spectrometry, and structural modeling to build a complete model of human RZZ. We find that RZZ is structurally related to self-assembling cytosolic coat scaffolds that mediate membrane cargo trafficking, including Clathrin, Sec13–Sec31, and αβ’ε-COP. We show that Spindly, a dynein adaptor, is related to BicD2 and binds RZZ directly in a farnesylation-dependent but membrane-independent manner. Through a targeted chemical biology approach, we identify ROD as the Spindly farnesyl receptor. Our results suggest that RZZ is dynein’s cargo at human kinetochores. PMID:28320825

Widespread Positive Selection Drives Differentiation of Centromeric Proteins in the Drosophila melanogaster subgroup.

PubMed

Beck, Emily A; Llopart, Ana

2015-11-25

Rapid evolution of centromeric satellite repeats is thought to cause compensatory amino acid evolution in interacting centromere-associated kinetochore proteins. Cid, a protein that mediates kinetochore/centromere interactions, displays particularly high amino acid turnover. Rapid evolution of both Cid and centromeric satellite repeats led us to hypothesize that the apparent compensatory evolution may extend to interacting partners in the Condensin I complex (i.e., SMC2, SMC4, Cap-H, Cap-D2, and Cap-G) and HP1s. Missense mutations in these proteins often result in improper centromere formation and aberrant chromosome segregation, thus selection for maintained function and coevolution among proteins of the complex is likely strong. Here, we report evidence of rapid evolution and recurrent positive selection in seven centromere-associated proteins in species of the Drosophila melanogaster subgroup, and further postulate that positive selection on these proteins could be a result of centromere drive and compensatory changes, with kinetochore proteins competing for optimal spindle attachment.

THE EFFECT OF UNRESOLVED BINARIES ON GLOBULAR CLUSTER PROPER-MOTION DISPERSION PROFILES

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bianchini, P.; Norris, M. A.; Ven, G. van de

2016-03-20

High-precision kinematic studies of globular clusters (GCs) require an accurate knowledge of all possible sources of contamination. Among other sources, binary stars can introduce systematic biases in the kinematics. Using a set of Monte Carlo cluster simulations with different concentrations and binary fractions, we investigate the effect of unresolved binaries on proper-motion dispersion profiles, treating the simulations like Hubble Space Telescope proper-motion samples. Since GCs evolve toward a state of partial energy equipartition, more-massive stars lose energy and decrease their velocity dispersion. As a consequence, on average, binaries have a lower velocity dispersion, since they are more-massive kinematic tracers. Wemore » show that, in the case of clusters with high binary fractions (initial binary fractions of 50%) and high concentrations (i.e., closer to energy equipartition), unresolved binaries introduce a color-dependent bias in the velocity dispersion of main-sequence stars of the order of 0.1–0.3 km s{sup −1} (corresponding to 1%−6% of the velocity dispersion), with the reddest stars having a lower velocity dispersion, due to the higher fraction of contaminating binaries. This bias depends on the ability to distinguish binaries from single stars, on the details of the color–magnitude diagram and the photometric errors. We apply our analysis to the HSTPROMO data set of NGC 7078 (M15) and show that no effect ascribable to binaries is observed, consistent with the low binary fraction of the cluster. Our work indicates that binaries do not significantly bias proper-motion velocity-dispersion profiles, but should be taken into account in the error budget of kinematic analyses.« less

"A Thousand Names They Called Him" Naming and Proper Names in the Work of S. Y. Agnon

ERIC Educational Resources Information Center

Hadad, Shira

2012-01-01

This dissertation offers a study of proper names and naming as a conceptual and thematic anchor in the work of S.Y. Agnon. Proper names, I argue, constitute an underexplored and highly fruitful prism through which to read literature, and specifically Agnon's fiction. My study consists of a series of readings in several of Agnon's major…

How to facilitate freshmen learning and support their transition to a university study environment

NASA Astrophysics Data System (ADS)

Kangas, Jari; Rantanen, Elisa; Kettunen, Lauri

2017-11-01

Most freshmen enter universities with high expectations and with good motivation, but too many are driven into performing instead of true learning. The issues are not only related to the challenge of comprehending the substance, social and other factors have an impact as well. All these multifaceted needs should be accounted for to facilitate student learning. Learning is an individual process and remarkable improvement in the learning practices is possible, if proper actions are addressed early enough. We motivate and describe a study of the experience obtained from a set of tailor-made courses that were given alongside standard curriculum. The courses aimed to provide a 'safe community' to address the multifaceted needs. Such support was integrated into regular coursework where active learning techniques, e.g. interactive small groups were incorporated. To assess impact of the courses we employ the feedback obtained during the courses and longitudinal statistical data about students' success.

Improvement of Accuracy of Proper Motions of Hipparcos Catalogue Stars Using Optical Latitude Observations

NASA Astrophysics Data System (ADS)

Damljanovic, G.

2009-09-01

Commission 19 (Earth Rotation) of the International Astronomical Union (IAU) established the Working Group on Earth Rotation in the Hipparcos Reference Frame (WG ERHRF) in 1995 to collect the optical observations of latitude and universal time variations, made during 1899.7 -- 1992.0 in line with the Earth orientation programmes (to derive Earth Orientation Parameters -- EOP), with Dr. Jan Vondrák (Astronomical Institute of Academy of Sciences of the Czech Republic, Prague) as the head of WG ERHRF. We participated in this international project using Belgrade Visual Zenith -- Telescope (BLZ) latitude data for the period 1949.0 -- 1986.0, after a new reduction of BLZ data made in my MSc thesis, finished in 1997 at the Faculty of Mathematics of University of Belgrade. Dr. Vondrák collected 4.4 million optical observations of latitude/universal time variations made at 33 observatories. The data were used for the EOP investigations, Hipparcos satellite Catalogue -- radio sources connection, etc. Nowadays, it is customary to correct the positions and proper motions of stars of Hipparcos Catalogue (as an optical reference frame) using ground -- based observations of some Hipparcos stars. In this PhD thesis we use the latitude observations made with several types of classical astrometric instruments: visual (ZT) and floating zenith -- telescope (FZT), visual zenith tube (VZT) and photographic zenith tube (PZT); 26 different instruments located at many observatories all over the world (used in the programs of monitoring the Earth orientation during the 20th century). We received the data from Dr. Vondrák via private communication. The observatories and instruments are: International Latitude Service -- ILS (Carloforte -- CA ZT, Cincinnati -- CI ZT, Gaithersburg -- GT ZT, Kitab -- KZ ZT, Mizusawa -- MZZ ZT, Tschardjui -- TS ZT and Ukiah -- UK ZT), Belgrade (BLZ ZT), Blagoveschtschensk (BK ZT), Irkutsk (IRZ ZT), Poltava (POL ZT), Pulkovo (PU and PUZ ZT), Varsovie (VJZ ZT

Calculating Proper Motions in the WFCAM Science Archive for the UKIRT Infrared Deep Sky Surveys

NASA Astrophysics Data System (ADS)

Collins, R.; Hambly, N.

2012-09-01

The ninth data release from the UKIRT Infrared Deep Sky Surveys (hereafter UKIDSS DR9), represents five years worth of observations by its wide-field camera (WFCAM) and will be the first to include proper motion values in its source catalogues for the shallow, wide-area surveys; the Large Area Survey (LAS), Galactic Clusters Survey (GCS) and (ultimately) Galactic Plane Survey (GPS). We, the Wide Field Astronomy Unit (WFAU) at the University of Edinburgh who prepare these regular data releases in the WFCAM Science Archive (WSA), describe in this paper how we make optimal use of the individual detection catalogues from each observation to derive high-quality astrometric fits for the positions of each detection enabling us to calculate a proper motion solution across multiple epochs and passbands when constructing a merged source catalogue. We also describe how the proper motion solutions affect the calculation of the various attributes provided in the database source catalogue tables, what measures of data quality we provide and a demonstration of the results for observations of the Pleiades cluster.

A Gaia-PS1-SDSS (GPS1) Proper Motion Catalog Covering 3/4 of the Sky

NASA Astrophysics Data System (ADS)

Tian, Hai-Jun; Gupta, Prashansa; Sesar, Branimir; Rix, Hans-Walter; Martin, Nicolas F.; Liu, Chao; Goldman, Bertrand; Platais, Imants; Kudritzki, Rolf-Peter; Waters, Christopher Z.

2017-09-01

We combine Gaia DR1, PS1, Sloan Digital Sky Survey (SDSS), and 2MASS astrometry to measure proper motions for 350 million sources across three-fourths of the sky down to a magnitude of {m}r˜ 20. Using positions of galaxies from PS1, we build a common reference frame for the multi-epoch PS1, single-epoch SDSS and 2MASS data, and calibrate the data in small angular patches to this frame. As the Gaia DR1 excludes resolved galaxy images, we choose a different approach to calibrate its positions to this reference frame: we exploit the fact that the proper motions of stars in these patches are linear. By simultaneously fitting the positions of stars at different epochs of—Gaia DR1, PS1, SDSS, and 2MASS—we construct an extensive catalog of proper motions dubbed GPS1. GPS1 has a characteristic systematic error of less than 0.3 {mas} {{yr}}-1 and a typical precision of 1.5-2.0 {mas} {{yr}}-1. The proper motions have been validated using galaxies, open clusters, distant giant stars, and QSOs. In comparison with other published faint proper motion catalogs, GPS1's systematic error (< 0.3 {mas} {{yr}}-1) should be nearly an order of magnitude better than that of PPMXL and UCAC4 (> 2.0 {mas} {{yr}}-1). Similarly, its precision (˜1.5 {mas} {{yr}}-1) is a four-fold improvement relative to PPMXL and UCAC4 (˜6.0 {mas} {{yr}}-1). For QSOs, the precision of GPS1 is found to be worse (˜2.0-3.0 {mas} {{yr}}-1), possibly due to their particular differential chromatic refraction. The GPS1 catalog will be released online and be available via the VizieR Service and VO Service.

Vantage Theory and the Use of English Demonstrative Determiners with Proper Nouns

ERIC Educational Resources Information Center

Riddle, Elizabeth M.

2010-01-01

This article discusses some apparently paradoxical behavior of the English demonstratives "this/these" and "that/those" as determiners of proper nouns and as metaphorical signals of epistemic and affective stance within the proximal-distal opposition. It is argued that the apparent paradoxes are actually cases of shifting perspectives or points of…

MERLIN observations of water maser proper motions in VY Canis Majoris

NASA Astrophysics Data System (ADS)

Richards, A. M. S.; Yates, J. A.; Cohen, R. J.

1998-09-01

MERLIN observations of the 22-GHz water masers in the circumstellar envelope of the supergiant VY CMa show an ellipsoidal distribution with a maximum extent of 700 mas east-west and 400 mas north-south. Comparison with observations made nine years earlier shows that the majority of maser features have survived and show proper motions throughout the region. The mean change in position is 28 mas and the proper motions are generally directed away from the assumed stellar position, and tend to be larger for features at greater projected distances. If the H_2O maser region is modelled as a partially filled thick spherical shell, and VY CMa is at a distance of 1.5 kpc, then the proper motion velocities in the direction of expansion are between 8kms^-1 at a distance of 75 mas from the assumed stellar position and 32kms^-1 at 360 mas. These velocities are consistent with the H_2O maser spectral line velocities which correspond to a maximum expansion velocity of 36kms^-1 at 400 mas from the assumed stellar position. These observations are consistent with radiation pressure on dust providing the force to accelerate the stellar wind as it passes through the H_2O maser shell. The H_2O maser region is elongated in the same direction as the dusty nebula around VY CMa. The water masers illuminate the small-scale dynamics and clumpiness which show the role of dust in driving the outflow. The overall ellipsoidal shape may be due to properties of the dust, such as its behaviour in the stellar magnetic field, or to interaction between the wind and circumstellar material. Maser monitoring also shows the difference between changes on the time-scale of stellar variability (a few years) and possible stages in the evolution of VY CMa to its likely fate as a supernova.

VizieR Online Data Catalog: The VLBA Extragalactic Proper Motion Catalog (Truebenbach+, 2017)

NASA Astrophysics Data System (ADS)

Truebenbach, A. E.; Darling, J.

2017-11-01

We created our catalog of extragalactic radio proper motions using the 2017a Goddard VLBI global solution. The 2017a solution is computed from more than 30 years of dual-band VLBI observations --1979 August 3 to 2017 March 27. We also observed 28 objects with either no redshift or a "questionable" Optical Characteristic of Astrometric Radio Sources (OCARS; Malkin 2016ARep...60..996M) redshift at the Apache Point Observatory (APO) 3.5m telescope and/or at Gemini North. We conducted observations on the 3.5m telescope at Apache Point Observatory with the Dual Imaging Spectrograph (DIS) from 2015 April 18 to 2016 June 30. We chose two objects for additional observations with the Gemini Multi-Object Spectrograph-North (GMOS-N) at Gemini North Observatory. 2021+317 was observed on 2016 June 26 and 28, while 0420+417 was observed on 2016 November 8 and 26. We also observed 42 radio sources with the Very Long Baseline Array (VLBA) in the X-band (3.6cm/8.3GHz). Our targets had all been previously observed by VLBI. Our VLBA observations were conducted in two campaigns from 2015 September to 2016 January and 2016 October to November. The final extragalactic proper motion catalog (created primarily from archival Goddard VLBI data, with redshifts obtained from OCARS) contains 713 proper motions with average uncertainties of 24μas/yr. (5 data files).

Review & Peer Review of “Parameters for Properly Designed and Operated Flares” Documents

EPA Pesticide Factsheets

This page contains two 2012 memoranda on the review of EPA's parameters for properly designed and operated flares. One details the process of peer review, and the other provides background information and specific charge questions to the panel.

Absolute proper motion of IRAS 00259+5625 with VERA: Indication of superbubble expansion motion

NASA Astrophysics Data System (ADS)

Sakai, Nobuyuki; Sato, Mayumi; Motogi, Kazuhito; Nagayama, Takumi; Shibata, Katsunori M.; Kanaguchi, Masahiro; Honma, Mareki

2014-02-01

We present the first measurement of the absolute proper motions of IRAS 00259+5625 (CB3, LBN594) associated with the H I loop called the "NGC 281 superbubble" that extends from the Galactic plane over ˜ 300 pc toward decreasing galactic latitude. The proper motion components measured with VLBI Exploration of Radio Astrometry (VERA) are (μαcos δ, μδ) = (-2.48 ± 0.32, -2.85 ± 0.65) mas yr-1, converted into (μlcos b, μb) = (-2.72 ± 0.32, -2.62 ± 0.65) mas yr-1 in the Galactic coordinates. The measured proper motion perpendicular to the Galactic plane (μb) shows vertical motion away from the Galactic plane with a significance of about ˜ 4 σ. As for the source distance, the distance measured with VERA is marginal, 2.4^{+1.0}_{-0.6} kpc. Using the distance, an absolute vertical motion (vb) of -17.9 ± 12.2 km s-1 is determined with ˜ 1.5 σ significance. The tendency towards the large vertical motion is consistent with previous very long baseline interferometry (VLBI) results for NGC 281 associated with the same superbubble. Thus, our VLBI results indicate superbubble expansion motion whose origin is believed to be sequential supernova explosions.

Oligonucleotide facilitators may inhibit or activate a hammerhead ribozyme.

PubMed Central

Jankowsky, E; Schwenzer, B

1996-01-01

Facilitators are oligonucleotides capable of affecting hammerhead ribozyme activity by interacting with the substrate at the termini of the ribozyme. Facilitator effects were determined in vitro using a system consisting of a ribozyme with 7 nucleotides in every stem sequence and two substrates with inverted facilitator binding sequences. The effects of 9mer and 12mer RNA as well as DNA facilitators which bind either adjacent to the 3'- or 5'-end of the ribozyme were investigated. A kinetic model was developed which allows determination of the apparent dissociation constant of the ribozyme-substrate complex from single turnover reactions. We observed a decreased dissociation constant of the ribozyme-substrate complex due to facilitator addition corresponding to an additional stabilization energy of delta delta G=-1.7 kcal/mol with 3'-end facilitators. The cleavage rate constant was increased by 3'-end facilitators and decreased by 5'-end facilitators. Values for Km were slightly lowered by all facilitators and kcat was increased by 3'-end facilitators and decreased by 5'-end facilitators in our system. Generally the facilitator effects increased with the length of the facilitators and RNA provided greater effects than DNA of the same sequence. Results suggest facilitator influences on several steps of the hammerhead reaction, substrate association, cleavage and dissociation of products. Moreover, these effects are dependent in different manners on ribozyme and substrate concentration. This leads to the conclusion that there is a concentration dependence whether activation or inhibition is caused by facilitators. Conclusions are drawn with regard to the design of hammerhead ribozyme facilitator systems. PMID:8602353

Facilitation of contrast detection in near-peripheral vision.

PubMed

Giorgi, Robert G; Soong, Grace P; Woods, Russell L; Peli, Eli

2004-12-01

Foveal detection of a Gabor patch (target) is facilitated by collinear, displaced high-contrast flankers. Polat and Sagi reported that the same phenomenon occurred in the periphery, but no data were presented [Proc. Natl. Acad. Sci. 91 (1994) 1206]. Others have found no facilitation in a limited number of conditions tested. To resolve this apparent conflict, we measured lateral facilitation in the near-periphery using a range of stimulus parameters. We found facilitation for a range of target-flanker distances for peripheral eccentricities up to 6 degrees , but the magnitude of the effect was less than found in central vision. Facilitation varied across subjects and with spatial frequency. Flanker contrast had no effect over the range evaluated (10-80%). Equal facilitation was found for two global arrangements of the stimulus pattern. Facilitation was found using a temporal, but not a spatial two-alternative forced-choice paradigm, accounting for the different results among previous studies. This finding supports previous indications of the role of attention in altering such facilitation. The value of facilitation from lateral interactions for persons with central vision impairment, who have to shift their attention to a peripheral locus constantly, needs to be examined.

Lowell proper motion survey: Southern Hemisphere (Giclas, Burnham, and Thomas 1978). Documentation for the machine-readable version

NASA Technical Reports Server (NTRS)

Warren, Wayne H., Jr.

1989-01-01

The machine-readable version of the catalog, as it is currently being distributed from the Astronomical Data Center, is described. The catalog is a summary compilation of the Lowell Proper Motion Survey for the Southern Hemisphere, as completed to mid-1978 and published in the Lowell Observatory Bulletins. This summary catalog serves as a Southern Hemisphere companion to the Lowell Proper Motion Survey, Northern Hemisphere.

The Proper Motion of the Central Compact Object RX J0822-4300 in the Supernova Remnant Puppis A

NASA Technical Reports Server (NTRS)

Becker, Werner; Prinz, Tobias; Winkler, P. Frank; Petre, Robert

2012-01-01

Using the High Resolution Camera (HRC) aboard the Chandra X-ray Observatory, we have re-examined the proper motion of the central compact object RX J0822-4300 in the supernova remnant Puppis A. New data from 2010 August, combined with three archival data sets from as early as 1999 December, provide a baseline of 3886 days (more than 10 1/2 years) to perform the measurement. Correlating the four positions of RX J0822-4300 measured in each data set implies a projected proper motion of mu = 71 +/- 12 mas/yr. For a distance of 2 kpc this proper motion is equivalent to a recoil velocity of 672 +/- 115 km/s. The position angle is found to be 244 +/- 11 degrees. Both the magnitude and direction of the proper motion are in agreement with RX J0822-4300 originating near the optical expansion center of the supernova remnant. For a displacement of 371 +/- 31 arcsec between its birth place and today's position we deduce an age of (5.2 +/- 1.0) 10(exp 3) yrs for RX J0822-4300. The age inferred from the neutron star proper motion and filament motions can be considered as two independent measurements of the same quantity. They average to 4450 +/- 750 yrs for the age of the supernova remnant Puppis A.

Primary Cilium-Regulated EG-VEGF Signaling Facilitates Trophoblast Invasion.

PubMed

Wang, Chia-Yih; Tsai, Hui-Ling; Syu, Jhih-Siang; Chen, Ting-Yu; Su, Mei-Tsz

2017-06-01

Trophoblast invasion is an important event in embryo implantation and placental development. During these processes, endocrine gland-derived vascular endothelial growth factor (EG-VEGF) is the key regulator mediating the crosstalk at the feto-maternal interface. The primary cilium is a cellular antenna receiving environmental signals and is crucial for proper development. However, little is known regarding the role of the primary cilium in early human pregnancy. Here, we demonstrate that EG-VEGF regulates trophoblast cell invasion via primary cilia. We found that EG-VEGF activated ERK1/2 signaling and subsequent upregulation of MMP2 and MMP9, thereby facilitating cell invasion in human trophoblast HTR-8/SVneo cells. Inhibition of ERK1/2 alleviated the expression of MMPs and trophoblast cell invasion after EG-VEGF treatment. In addition, primary cilia were observed in all the trophoblast cell lines tested and, more importantly, in human first-trimester placental tissue. The receptor of EG-VEGF, PROKR1, was detected in primary cilia. Depletion of IFT88, the intraflagellar transporter required for ciliogenesis, inhibited primary cilium growth, thereby ameliorating ERK1/2 activation, MMP upregulation, and trophoblast cell invasion promoted by EG-VEGF. These findings demonstrate a novel function of primary cilia in controlling EG-VEGF-regulated trophoblast invasion and reveal the underlying molecular mechanism. J. Cell. Physiol. 232: 1467-1477, 2017. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

A Study on How to Breathe Properly When Practicing Tai Chi Chuan

ERIC Educational Resources Information Center

Yang, Hanchun

2011-01-01

When practicing Tai Chi Chuan, proper breath plays an important role in shaping Tai Chi Chuan's style and its fitness value. The paper aims to analyse the postures of Tai Chi Chuan and its breath characteristics. The paper also presents some new insights on how to co-ordinate breath with postures by case studies.

Proper motion separation of Be stars in the Milky Way and the Magellanic Clouds

NASA Astrophysics Data System (ADS)

Vieira, K.; García, A.; Sabogal, B.

2018-01-01

We present a proper motion investigation of a sample of Be stars candidates towards the Large Magellanic Cloud (LMC), which has resulted in the identification of two separate populations, in the Galactic foreground and in the Magellanic background. OGLE BVI and 2MASS JHK photometry were used with the SPM4 proper motions to discriminate the different populations located towards the LMC. Two populations with distinctive infrared colours and noticeable different kinematics were found, the bluer sample is consistent with being in the LMC and the redder one with belonging to the Milky Way (MW) disk. This settles the nature of the redder sample which had been described in previous publications as a possible unknown subclass of stars among the Be candidates in the LMC.

Determination of Proper Austenitization Temperatures for Hot Stamping of AISI 4140 Steel

NASA Astrophysics Data System (ADS)

Samadian, Pedram; Parsa, Mohammad Habibi; Shakeri, Amid

2014-04-01

High strength steels are desirable materials for use in automobile bodies in order to reduce vehicle weight and increase the safety of car passengers, but steel grades with high strength commonly show poor formability. Recently, steels with controlled microstructures and compositions are used to gain adequate strength after hot stamping while maintaining good formability during processing. In this study, microstructure evolutions and changes in mechanical properties of AISI 4140 steel sheets resulting from the hot stamping process at different austenitization temperatures were investigated. To determine the proper austenitization temperatures, the results were compared with those of the cold-worked and cold-worked plus quench-tempered specimens. Comparisons showed that the austenitization temperatures of 1000 and 1100 °C are proper for hot stamping of 3-mm-thick AISI 4140 steel sheets due to the resultant martensitic microstructure which led to the yield and ultimate tensile strength of 1.3 and 2.1 GPa, respectively. Such conditions resulted in more favorable simultaneous strength and elongation than those of hot-stamped conventional boron steels.

Analyzing the Proper Motion of Two Double Star Systems from Astrometric Measurements

NASA Astrophysics Data System (ADS)

Falatoun, Alex; Barrera, Janet; de Neef, Anna; Gonzalez, Aura; Calanog, Jae; Boyce, Pat; Boyce, Grady

2018-04-01

The iTelescope network was used to obtain astrometric measurements of double star systems WDS 12202-1408 (STF 1631) and WDS 12339+5522 (STI 2286). Through astrometric measurement softwares SAOImage DS9 and Mira Pro x64, a mean position angle for STF 1631 of 304.8° ± 0.9° and a mean separation 14.7" ± 0.2" was measured. For STI 2286, a newly measured mean position angle of 85.9° ± 0.9° and mean separation 11.5" ± 0.3" were obtained. The relative proper motion of 1631 shows that the system could be demonstrating a linear path or an approximately circular orbit with a period of 1400 years. Parallax measurements of the secondary star will aid in classifying if this system is a physical or a visual pair. The proper motion of STI 2286 indicates that it could be a physical pair, featuring an orbit nearing a turning point. Follow-up observations in three to four year intervals will further validate or refute this claim and constrain the shape of a possible orbit.

Technologies and Techniques for Supporting Facilitated Video

ERIC Educational Resources Information Center

Linnell, Natalie

2011-01-01

Worldwide, demand for education of all kinds is increasing beyond the capacity to provide it. One approach that shows potential for addressing this demand is facilitated video. In facilitated video, an educator is recorded teaching, and that video is sent to a remote site where it is shown to students by a facilitator who creates interaction…

Transducer model produces facilitation from opposite-sign flanks

NASA Technical Reports Server (NTRS)

Solomon, J. A.; Watson, A. B.; Morgan, M. J.

1999-01-01

Small spots, lines and Gabor patterns can be easier to detect when they are superimposed upon similar spots, lines and Gabor patterns. Traditionally, such facilitation has been understood to be a consequence of nonlinear contrast transduction. Facilitation has also been reported to arise from non-overlapping patterns with opposite sign. We point out that this result does not preclude the traditional explanation for superimposed targets. Moreover, we find that facilitation from opposite-sign flanks is weaker than facilitation from same-sign flanks. Simulations with a transducer model produce opposite-sign facilitation.

What can we learn from facilitator and student perceptions of facilitation skills and roles in the first year of a problem-based learning curriculum?

PubMed Central

McLean, Michelle

2003-01-01

Background The small group tutorial is a cornerstone of problem-based learning. By implication, the role of the facilitator is of pivotal importance. The present investigation canvassed perceptions of facilitators with differing levels of experience regarding their roles and duties in the tutorial. Methods In January 2002, one year after problem-based learning implementation at the Nelson R. Mandela School of Medicine, facilitators with the following experience were canvassed: trained and about to facilitate, facilitated once only and facilitated more than one six-week theme. Student comments regarding facilitator skills were obtained from a 2001 course survey. Results While facilitators generally agreed that the three-day training workshop provided sufficient insight into the facilitation process, they become more comfortable with increasing experience. Many facilitators experienced difficulty not providing content expertise. Again, this improved with increasing experience. Most facilitators saw students as colleagues. They agreed that they should be role models, but were less enthusiastic about being mentors. Students were critical of facilitators who were not up to date with curriculum implementation or who appeared disinterested. While facilitator responses suggest that there was considerable intrinsic motivation, this might in fact not be the case. Conclusions Even if they had facilitated on all six themes, facilitators could still be considered as novices. Faculty support is therefore critical for the first few years of problem-based learning, particularly for those who had facilitated once only. Since student and facilitator expectations in the small group tutorial may differ, roles and duties of facilitators must be explicit for both parties from the outset. PMID:14585108

What can we learn from facilitator and student perceptions of facilitation skills and roles in the first year of a problem-based learning curriculum?

PubMed

McLean, Michelle

2003-10-30

The small group tutorial is a cornerstone of problem-based learning. By implication, the role of the facilitator is of pivotal importance. The present investigation canvassed perceptions of facilitators with differing levels of experience regarding their roles and duties in the tutorial. In January 2002, one year after problem-based learning implementation at the Nelson R. Mandela School of Medicine, facilitators with the following experience were canvassed: trained and about to facilitate, facilitated once only and facilitated more than one six-week theme. Student comments regarding facilitator skills were obtained from a 2001 course survey. While facilitators generally agreed that the three-day training workshop provided sufficient insight into the facilitation process, they become more comfortable with increasing experience. Many facilitators experienced difficulty not providing content expertise. Again, this improved with increasing experience. Most facilitators saw students as colleagues. They agreed that they should be role models, but were less enthusiastic about being mentors. Students were critical of facilitators who were not up to date with curriculum implementation or who appeared disinterested. While facilitator responses suggest that there was considerable intrinsic motivation, this might in fact not be the case. Even if they had facilitated on all six themes, facilitators could still be considered as novices. Faculty support is therefore critical for the first few years of problem-based learning, particularly for those who had facilitated once only. Since student and facilitator expectations in the small group tutorial may differ, roles and duties of facilitators must be explicit for both parties from the outset.

20 CFR 1002.307 - What is the proper venue in an action against a State or private employer?

Code of Federal Regulations, 2010 CFR

2010-04-01

... 20 Employees' Benefits 3 2010-04-01 2010-04-01 false What is the proper venue in an action against a State or private employer? 1002.307 Section 1002.307 Employees' Benefits OFFICE OF THE ASSISTANT... Enforcement of Rights and Benefits Against A State Or Private Employer § 1002.307 What is the proper venue in...

“Real-Time” Cosmology with Extragalactic Proper Motions: the Secular Aberration Drift and Evolution of Large-Scale Structure

NASA Astrophysics Data System (ADS)

Truebenbach, Alexandra; Darling, Jeremy

2018-01-01

We present the VLBA Extragalactic Proper Motion Catalog, a catalog of extragalactic proper motions created using archival VLBI data and our own VLBA astrometry. The catalog contains 713 proper motions, with average uncertainties of ~ 24 microarcsec/yr, including 40 new or improved proper motion measurements using relative astrometry with the VLBA. We detect the secular aberration drift – the apparent motion of extragalactic objects caused by the solar system's acceleration around the Galactic Center – at 6.3 sigma significance with an amplitude of 1.69 +/- 0.27 microarcsec/yr and an apex consistent with the Galactic Center (275.2 +/- 10.0 deg, -29.4 +/- 8.8 deg). Our dipole model detects the aberration drift at a higher significance than some previous studies (e.g., Titov & Lambert 2013), but at a lower amplitude than expected or previously measured. We then use the correlated relative proper motions of extragalactic objects to place upper limits on the rate of large-scale structure collapse (e.g., Quercellini et al. 2009; Darling 2013). Pairs of small separation objects that are in gravitationally interacting structures such as filaments of large-scale structure will show a net decrease in angular separation (> - 15.5 microarcsec/yr) as they move towards each other, while pairs of large separation objects that are gravitationally unbound and move with the Hubble expansion will show no net change in angular separation. With our catalog, we place a 3 sigma limit on the rate of convergence of large-scale structure of -11.4 microarcsec/yr for extragalactic objects within 100 comoving Mpc of each other. We also confirm that large separation objects (> 800 comoving Mpc) move with the Hubble flow to within ~ 2.2 microarcsec/yr. In the future, we plan to incorporate the upcoming Gaia proper motions into our catalog to achieve a higher precision measurement of the average relative proper motion of gravitationally interacting extragalactic objects and to refine our

Sense of Accomplishment Is Modulated by a Proper Level of Instruction and Represented in the Brain Reward System.

PubMed

Nakai, Tomoya; Nakatani, Hironori; Hosoda, Chihiro; Nonaka, Yulri; Okanoya, Kazuo

2017-01-01

Problem-solving can be facilitated with instructions or hints, which provide information about given problems. The proper amount of instruction that should be provided for learners is controversial. Research shows that tasks with intermediate difficulty induce the largest sense of accomplishment (SA), leading to an intrinsic motivation for learning. To investigate the effect of instructions, we prepared three instruction levels (No hint, Indirect hint, and Direct hint) for the same insight-problem types. We hypothesized that indirect instructions impose intermediate difficulty for each individual, thereby inducing the greatest SA per person. Based on previous neuroimaging studies that showed involvement of the bilateral caudate in learning and motivation, we expected SA to be processed in this reward system. We recruited twenty-one participants, and investigated neural activations during problem solving by functional magnetic resonance imaging (fMRI). We confirmed that the Indirect hint, which imposed intermediate difficulty, induced the largest SA among the three instruction types. Using fMRI, we showed that activations in the bilateral caudate and anterior cingulate cortex (ACC) were significantly modulated by SA. In the bilateral caudate, the indirect hint induced the largest activation, while the ACC seemed to reflect the difference between correct and incorrect trials. Importantly, such activation pattern was independent of notations (number or letter). Our results indicate that SA is represented in the reward system, and that the Indirect instruction effectively induces such sensation.

Translating Proper Nouns: A Case Study on English Translation of Hafez's Lyrics

ERIC Educational Resources Information Center

Shirinzadeh, Seyed Alireza; Mahadi, Tengku Sepora Tengku

2014-01-01

Proper nouns are regarded so simple that they might be taken for granted in translation explorations. Some may believe that they should not be translated in transmitting source texts to target texts. But, it is not the case; if one looks at present translations, he will notice that different strategies might be applied for translating proper…

Astrometry in the globular cluster M13. II. Membership probabilities from old proper motions

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cudworth, K.

Astrometric cluster membership probabilities have been derived from proper motions measured by other authors for stars in the region of the globular cluster M13. Several stars of individual interest are discussed.

The VMC survey. XXX. Stellar proper motions in the central parts of the Small Magellanic Cloud

NASA Astrophysics Data System (ADS)

Niederhofer, F.; Cioni, M.-R. L.; Rubele, S.; Schmidt, T.; Bekki, K.; Grijs, R. de; Emerson, J.; Ivanov, V. D.; Marconi, M.; Oliveira, J. M.; Petr-Gotzens, M. G.; Ripepi, V.; van Loon, J. Th.; Zaggia, S.

2018-05-01

We present the first spatially resolved map of stellar proper motions within the central ( 3.1 × 2.4 kpc) regions of the Small Magellanic Cloud (SMC). The data used for this study encompasses four tiles from the ongoing near-infrared VISTA survey of the Magellanic Clouds system and covers a total contiguous area on the sky of 6.81 deg2. Proper motions have been calculated independently in two dimensions from the spatial offsets in the Ks filter over time baselines between 22 and 27 months. The reflex motions of approximately 33 000 background galaxies are used to calibrate the stellar motions to an absolute scale. The resulting catalog is composed of more than 690 000 stars which have been selected based on their position in the (J - Ks, Ks) color-magnitude diagram. For the median absolute proper motion of the SMC, we find (μαcos(δ), μδ) = (1.087 ± 0.192 (sys.) ± 0.003 (stat.), -1.187 ± 0.008 (sys.) ± 0.003 (stat.)) mas yr-1, consistent with previous studies. Mapping the proper motions as a function of position within the SMC reveals a nonuniform velocity pattern indicative of a tidal feature behind the main body of the SMC and a flow of stars in the south-east moving predominantly along the line-of-sight. Based on observations made with VISTA at the Paranal Observatory under program ID 179.B-2003.

Facilitator training program: The Université Laval Interprofessional Initiative.

PubMed

Milot, Élise; Museux, Anne-Claire; Careau, Emmanuelle

2017-03-01

A facilitator training program (FTP) for interprofessional learning (IPL) facilitators has been developed at Université Laval. This article describes the impacts of this program as perceived by the 22 IPL facilitators involved and outlines recommendations. Two qualitative data collection strategies were used to document the facilitators' pedagogical needs and views of the program's impacts. Results suggest that the FTP's pedagogical approach was effective. The IPL facilitators became more aware of their challenges and identified concrete strategies to use. Training initiatives should equip IPL facilitators to cope with uncertainty, create a climate supporting active learning, and facilitate positive interactions between students.

49 CFR 38.2 - Equivalent facilitation.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 49 Transportation 1 2010-10-01 2010-10-01 false Equivalent facilitation. 38.2 Section 38.2 Transportation Office of the Secretary of Transportation AMERICANS WITH DISABILITIES ACT (ADA) ACCESSIBILITY SPECIFICATIONS FOR TRANSPORTATION VEHICLES General § 38.2 Equivalent facilitation. Departures from particular...

A focused educational intervention can promote the proper application of seat belts during pregnancy.

PubMed

McGwin, Gerald; Willey, Paula; Ware, Amy; Kohler, Connie; Kirby, Tyler; Rue, Loring W

2004-05-01

The failure to properly use automobile restraints during pregnancy is a significant contributor to both maternal and fetal injury and death. Misconceptions as to the effects of restraint use on the fetus and a lack of instruction as to proper restraint positioning contribute to this problem. Focus groups were used to develop an intervention consisting of educational material for prenatal care clinic patients and staff pertaining to seat belt use during pregnancy. The intervention was administered over a 2-month period. Two groups of women, one preintervention and one postintervention, were surveyed to determine demographics, pregnancy status, and current restraint use characteristics. Preintervention and postintervention surveys were administered to 450 and 285 women, respectively. The proportion of women reporting correct placement of seat belts increased from 70.8% to 83.0% (p < 0.001) after the intervention. Knowledge of seat belt effectiveness also increased significantly (p < 0.001) after the intervention. Only 25.2% of women in the preintervention group reported receiving information from clinic staff on restraint use compared with 76.8% of the postintervention group (p < 0.001). Lack of knowledge regarding restraint use during pregnancy contributes to a lack of consistent and proper use of restraints. Educational tools that improve both knowledge and behavior have the potential to increase automotive safety during pregnancy.

The proper role of the cost-of-equity concept in pragmatic utility regulation

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kihm, Steven G.

2007-12-15

As classic treatises make clear, determining a reasonable return on equity is a judgment call, one that reflect the regulator's broad perspective on public policy matters. That requires one to look beyond economic concepts, such as cost of equity, to find the proper return. (author)

SLoWPoKES-II: 100,000 WIDE BINARIES IDENTIFIED IN SDSS WITHOUT PROPER MOTIONS

DOE Office of Scientific and Technical Information (OSTI.GOV)

Dhital, Saurav; West, Andrew A.; Schluns, Kyle J.

2015-08-15

We present the Sloan Low-mass Wide Pairs of Kinematically Equivalent Stars (SLoWPoKES)-II catalog of low-mass visual binaries identified from the Sloan Digital Sky Survey (SDSS) by matching photometric distances. The candidate pairs are vetted by comparing the stellar information. The candidate pairs are vetted by comparing the stellar density at their respective Galactic positions to Monte Carlo realizations of a simulated Milky Way. In this way, we are able to identify large numbers of bona fide wide binaries without the need for proper motions. Here, 105,537 visual binaries with angular separations of ∼1–20″ were identified, each with a probability ofmore » chance alignment of ≤5%. This is the largest catalog of bona fide wide binaries to date, and it contains a diversity of systems—in mass, mass ratios, binary separations, metallicity, and evolutionary states—that should facilitate follow-up studies to characterize the properties of M dwarfs and white dwarfs. There is a subtle but definitive suggestion of multiple populations in the physical separation distribution, supporting earlier findings. We suggest that wide binaries are composed of multiple populations, most likely representing different formation modes. There are 141 M7 or later wide binary candidates, representing a seven-fold increase over the number currently known. These binaries are too wide to have been formed via the ejection mechanism. Finally, we found that 6% of spectroscopically confirmed M dwarfs are not included in the SDSS STAR catalog; they are misclassified as extended sources due to the presence of a nearby or partially resolved companion. The SLoWPoKES-II catalog is publicly available to the entire community on the World Wide Web via the Filtergraph data visualization portal.« less

An exploration of tutors' experiences of facilitating problem-based learning. Part 2--implications for the facilitation of problem based learning.

PubMed

Haith-Cooper, Melanie

2003-01-01

This paper is the second of two parts exploring a study that was undertaken to investigate the role of the tutor in facilitating problem-based learning (PBL). The first part focussed on the methodological underpinnings of the study. This paper aims to focus on the findings of the study and their implications for the facilitation of PBL. Six essential themes emerged from the findings that described the facilitation role. The tutors believed that their facilitation role was essentially structured around the decision of when to intervene and how to intervene in the PBL process. Modelling and non-verbal communication were seen as essential strategies for the facilitator. Underpinning these decisions was the need to trust in the philosophy of PBL. However, within many of the themes, there was a divergence of opinion as to how the role should actually be undertaken. Despite this, these findings have implications for the future role of PBL facilitators in Health Professional Education.

49 CFR 392.64 - Riding within closed commercial motor vehicles without proper exits.

Code of Federal Regulations, 2014 CFR

2014-10-01

... 49 Transportation 5 2014-10-01 2014-10-01 false Riding within closed commercial motor vehicles without proper exits. 392.64 Section 392.64 Transportation Other Regulations Relating to Transportation (Continued) FEDERAL MOTOR CARRIER SAFETY ADMINISTRATION, DEPARTMENT OF TRANSPORTATION FEDERAL MOTOR CARRIER SAFETY REGULATIONS DRIVING OF COMMERCIAL...

Rationale, design features, and baseline characteristics: The Heart Institute of Japan-PRoper level of lipid lOwering with Pitavastatin and Ezetimibe in acute coRonary syndrome (HIJ-PROPER).

PubMed

Kawada-Watanabe, Erisa; Ogawa, Hiroshi; Koyanagi, Ryo; Arashi, Hiroyuki; Yamaguchi, Junichi; Matsui, Kunihiko; Hagiwara, Nobuhisa

2017-03-01

In contrast to current guidelines in Western countries, moderate reduction of low-density lipoprotein cholesterol (LDL-C) is recommended for Japanese patients with atherosclerotic cardiovascular disease and dyslipidemia even in secondary prevention. HIJ-PROPER (Heart Institute of Japan-PRoper level of lipid lOwering with Pitavastatin and Ezetimibe in acute coRonary syndrome) is a prospective, randomized, open-label, blinded endpoint multicenter trial designed to assess whether closely controlled LDL-C lowering with a standard statin dose plus ezetimibe, targeting LDL-C of <70mg/dL, would reduce cardiovascular events more than standard statin monotherapy targeting LDL-C of <100mg/dL as per the Japan Atherosclerotic Society guideline in patients with acute coronary syndrome (ACS) and dyslipidemia. We recruited patients with ACS and dyslipidemia who had undergone coronary angiography. Participants are randomly allocated to either intensive LDL-C lowering treatment (target LDL-C of <70mg/dL; pitavastatin plus ezetimibe) or standard LDL-C lowering treatment (target LDL-C of 90-100mg/dL; pitavastatin monotherapy). The primary endpoint is a composite of total death, non-fatal myocardial infarction (MI), non-fatal stroke, unstable angina, and any ischemia-driven revascularization. Patients will be followed for a minimum of 3 years. Between January 2010 and April 2013, 1734 patients were enrolled from 19 hospitals in Japan with a mean age of 65.6 years; 75.5% were men and 83.3% were statin-naïve. The qualifying ACS was an acute MI in 61.5%. This study is expected to report its findings in August 2016. HIJ-PROPER will determine whether targeting LDL-C of <70mg/dL with pitavastatin plus ezetimibe can improve cardiovascular outcomes in Japanese patients with ACS and dyslipidemia in comparison to targeting LDL-C of 90-100mg/dL with standard pitavastatin monotherapy. UMIN000002742. Copyright © 2016 Japanese College of Cardiology. Published by Elsevier Ltd. All rights reserved.

On the systematics in apparent proper motions of radio sources observed by VLBI

NASA Astrophysics Data System (ADS)

Raposo-Pulido, V.; Lambert, S.; Capitaine, N.; Nilsson, T.; Heinkelmann, R.; Schuh, H.

2015-08-01

For about twenty years, several authors have been investigating the systematics in the apparent proper motions of radio source positions. In some cases, the theoretical work developed (Pyne et al., 1996) could not be assessed due to the few number of VLBI observations. In other cases, the effects attributed to apparent proper motion could not be related successfully because there were no significant evidences from a statistical point of view (MacMillan, 2005). In this work we provide considerations about the estimation of the coefficients of spherical harmonics, based on a three-step procedure used by Titov et al. (2011) and Titov and Lambert (2013). The early stage of this work has been to compare step by step the computations and estimation processes between the Calc/Solve (http://gemini.gsfc.nasa.gov/solve/) and VieVS software (Böhm et al., 2012). To achieve this, the results were analyzed and compared with the previous study done by Titov and Lambert (2013).

PAK4 kinase is essential for embryonic viability and for proper neuronal development.

PubMed

Qu, Jian; Li, Xiaofan; Novitch, Bennet G; Zheng, Ye; Kohn, Matthew; Xie, Jian-Ming; Kozinn, Spencer; Bronson, Roderick; Beg, Amer A; Minden, Audrey

2003-10-01

The serine/threonine kinase PAK4 is a target for the Rho GTPase Cdc42 and has been shown to regulate cell morphology and cytoskeletal organization in mammalian cells. To examine the physiological and developmental functions of PAK4, we have disrupted the PAK4 gene in mice. The absence of PAK4 led to lethality by embryonic day 11.5, a result most likely due to a defect in the fetal heart. Striking abnormalities were also evident in the nervous systems of PAK4-deficient embryos. These embryos had dramatic defects in neuronal development and axonal outgrowth. In particular, spinal cord motor neurons and interneurons failed to differentiate and migrate to their proper positions. This is probably related to the role for PAK4 in the regulation of cytoskeletal organization and cell and/or extracellular matrix adhesion. PAK4-null embryos also had defects in proper folding of the caudal portion of the neural tube, suggesting an important role for PAK4 in neural tube development.

An adaptive model order reduction by proper snapshot selection for nonlinear dynamical problems

NASA Astrophysics Data System (ADS)

Nigro, P. S. B.; Anndif, M.; Teixeira, Y.; Pimenta, P. M.; Wriggers, P.

2016-04-01

Model Order Reduction (MOR) methods are employed in many fields of Engineering in order to reduce the processing time of complex computational simulations. A usual approach to achieve this is the application of Galerkin projection to generate representative subspaces (reduced spaces). However, when strong nonlinearities in a dynamical system are present and this technique is employed several times along the simulation, it can be very inefficient. This work proposes a new adaptive strategy, which ensures low computational cost and small error to deal with this problem. This work also presents a new method to select snapshots named Proper Snapshot Selection (PSS). The objective of the PSS is to obtain a good balance between accuracy and computational cost by improving the adaptive strategy through a better snapshot selection in real time (online analysis). With this method, it is possible a substantial reduction of the subspace, keeping the quality of the model without the use of the Proper Orthogonal Decomposition (POD).

Familiarity facilitates feature-based face processing.

PubMed

Visconti di Oleggio Castello, Matteo; Wheeler, Kelsey G; Cipolli, Carlo; Gobbini, M Ida

2017-01-01

Recognition of personally familiar faces is remarkably efficient, effortless and robust. We asked if feature-based face processing facilitates detection of familiar faces by testing the effect of face inversion on a visual search task for familiar and unfamiliar faces. Because face inversion disrupts configural and holistic face processing, we hypothesized that inversion would diminish the familiarity advantage to the extent that it is mediated by such processing. Subjects detected personally familiar and stranger target faces in arrays of two, four, or six face images. Subjects showed significant facilitation of personally familiar face detection for both upright and inverted faces. The effect of familiarity on target absent trials, which involved only rejection of unfamiliar face distractors, suggests that familiarity facilitates rejection of unfamiliar distractors as well as detection of familiar targets. The preserved familiarity effect for inverted faces suggests that facilitation of face detection afforded by familiarity reflects mostly feature-based processes.

Facilitating LOS Debriefings: A Training Manual

NASA Technical Reports Server (NTRS)

McDonnell, Lori K.; Jobe, Kimberly K.; Dismukes, R. Key

1997-01-01

This manual is a practical guide to help airline instructors effectively facilitate debriefings of Line Oriented Simulations (LOS). It is based on a recently completed study of Line Oriented Flight Training (LOFT) debriefings at several U.S. airlines. This manual presents specific facilitation tools instructors can use to achieve debriefing objectives. The approach of the manual is to be flexible so it can be tailored to the individual needs of each airline. Part One clarifies the purpose and objectives of facilitation in the LOS setting. Part Two provides recommendations for clarifying roles and expectations and presents a model for organizing discussion. Part Tree suggests techniques for eliciting active crew participation and in-depth analysis and evaluation. Finally, in Part Four, these techniques are organized according to the facilitation model. Examples of how to effectively use the techniques are provided throughout, including strategies to try when the debriefing objectives are not being fully achieved.

The Optical Gravitational Lensing Experiment: catalogue of stellar proper motions in the OGLE-II Galactic bulge fields

NASA Astrophysics Data System (ADS)

Sumi, T.; Wu, X.; Udalski, A.; Szymański, M.; Kubiak, M.; Pietrzyński, G.; Soszyński, I.; Woźniak, P.; Żebruń, K.; Szewczyk, O.; Wyrzykowski, Ł.

2004-03-01

We present a proper-motion (μ) catalogue of 5 080 236 stars in 49 Optical Gravitational Lensing Experiment II (OGLE-II) Galactic bulge (GB) fields, covering a range of -11° < l < 11° and -6° < b < 3°, the total area close to 11 deg2. The proper-motion measurements are based on 138-555 I-band images taken during four observing seasons: 1997-2000. The catalogue stars are in the magnitude range 11 < I < 18 mag. In particular, the catalogue includes red clump giants and red giants in the GB, and main-sequence stars in the Galactic disc. The proper motions up to μ= 500 mas yr-1 were measured with a mean accuracy of 0.8-3.5 mas yr-1, depending on the brightness of a star. This catalogue may be useful for studying the kinematics of stars in the GB and the Galactic disc.

Proper and improper zero energy modes in Hartree-Fock theory and their relevance for symmetry breaking and restoration.

PubMed

Cui, Yao; Bulik, Ireneusz W; Jiménez-Hoyos, Carlos A; Henderson, Thomas M; Scuseria, Gustavo E

2013-10-21

We study the spectra of the molecular orbital Hessian (stability matrix) and random-phase approximation (RPA) Hamiltonian of broken-symmetry Hartree-Fock solutions, focusing on zero eigenvalue modes. After all negative eigenvalues are removed from the Hessian by following their eigenvectors downhill, one is left with only positive and zero eigenvalues. Zero modes correspond to orbital rotations with no restoring force. These rotations determine states in the Goldstone manifold, which originates from a spontaneously broken continuous symmetry in the wave function. Zero modes can be classified as improper or proper according to their different mathematical and physical properties. Improper modes arise from symmetry breaking and their restoration always lowers the energy. Proper modes, on the other hand, correspond to degeneracies of the wave function, and their symmetry restoration does not necessarily lower the energy. We discuss how the RPA Hamiltonian distinguishes between proper and improper modes by doubling the number of zero eigenvalues associated with the latter. Proper modes in the Hessian always appear in pairs which do not double in RPA. We present several pedagogical cases exemplifying the above statements. The relevance of these results for projected Hartree-Fock methods is also addressed.

Constraint of gene expression by the chromatin remodelling protein CHD4 facilitates lineage specification

PubMed Central

O'Shaughnessy-Kirwan, Aoife; Signolet, Jason; Costello, Ita; Gharbi, Sarah; Hendrich, Brian

2015-01-01

Chromatin remodelling proteins are essential for different aspects of metazoan biology, yet functional details of why these proteins are important are lacking. Although it is possible to describe the biochemistry of how they remodel chromatin, their chromatin-binding profiles in cell lines, and gene expression changes upon loss of a given protein, in very few cases can this easily translate into an understanding of how the function of that protein actually influences a developmental process. Here, we investigate how the chromatin remodelling protein CHD4 facilitates the first lineage decision in mammalian embryogenesis. Embryos lacking CHD4 can form a morphologically normal early blastocyst, but are unable to successfully complete the first lineage decision and form functional trophectoderm (TE). In the absence of a functional TE, Chd4 mutant blastocysts do not implant and are hence not viable. By measuring transcript levels in single cells from early embryos, we show that CHD4 influences the frequency at which unspecified cells in preimplantation stage embryos express lineage markers prior to the execution of this first lineage decision. In the absence of CHD4, this frequency is increased in 16-cell embryos, and by the blastocyst stage cells fail to properly adopt a TE gene expression programme. We propose that CHD4 allows cells to undertake lineage commitment in vivo by modulating the frequency with which lineage-specification genes are expressed. This provides novel insight into both how lineage decisions are made in mammalian cells, and how a chromatin remodelling protein functions to facilitate lineage commitment. PMID:26116663

40 CFR 85.2104 - Owners' compliance with instructions for proper maintenance and use.

Code of Federal Regulations, 2011 CFR

2011-07-01

... automobiles for the relevant maintenance instruction(s); or (2) A showing that the vehicle has been submitted... to someone who regularly engages in the business of servicing automobiles for the purpose of... area in which the vehicle or engine is located, unless the written instructions for proper maintenance...

40 CFR 85.2104 - Owners' compliance with instructions for proper maintenance and use.

Code of Federal Regulations, 2013 CFR

2013-07-01

... automobiles for the relevant maintenance instruction(s); or (2) A showing that the vehicle has been submitted... to someone who regularly engages in the business of servicing automobiles for the purpose of... area in which the vehicle or engine is located, unless the written instructions for proper maintenance...

40 CFR 85.2104 - Owners' compliance with instructions for proper maintenance and use.

Code of Federal Regulations, 2014 CFR

2014-07-01

... automobiles for the relevant maintenance instruction(s); or (2) A showing that the vehicle has been submitted... to someone who regularly engages in the business of servicing automobiles for the purpose of... area in which the vehicle or engine is located, unless the written instructions for proper maintenance...

40 CFR 85.2104 - Owners' compliance with instructions for proper maintenance and use.

Code of Federal Regulations, 2010 CFR

2010-07-01

... automobiles for the relevant maintenance instruction(s); or (2) A showing that the vehicle has been submitted... to someone who regularly engages in the business of servicing automobiles for the purpose of... area in which the vehicle or engine is located, unless the written instructions for proper maintenance...

40 CFR 85.2104 - Owners' compliance with instructions for proper maintenance and use.

Code of Federal Regulations, 2012 CFR

2012-07-01

... automobiles for the relevant maintenance instruction(s); or (2) A showing that the vehicle has been submitted... to someone who regularly engages in the business of servicing automobiles for the purpose of... area in which the vehicle or engine is located, unless the written instructions for proper maintenance...

Barriers and enablers to proper diabetic foot care amongst community dwellers in an Asian population: a qualitative study

PubMed Central

Cuttilan, Amit Nirmal; Pearce, Christopher J.

2017-01-01

Background Diabetic foot complications are common within Asian populations. They arise due to poor diabetic control and foot care. In this study, we aimed to identify the causes for proper and improper diabetic foot care. Methods A qualitative study consisting of key informant interviews with 17 healthcare professionals, including doctors and various allied health workers, was conducted. Participants included had at least five years of caring for diabetic foot patients either in public institutions or private clinics. Data collected was analyzed via thematic analysis. Results Diabetic patients were generally observed to have a mixture of proper and improper information and beliefs which eventually resulted in the extent of proper foot care. Factors which influenced the extent of proper and improper state of information and beliefs were classified into predisposing and precipitating factors. Predisposing factors were further categorised into modifiable factors (e.g., education level, socioeconomic status, social support) and non-modifiable factors (e.g., age, presence and severity of co-morbidities restricting ability to selfcare, past experiences). Precipitating factors were categorized into patient factors (e.g., degree of reception of information, presence of psychological barriers), provider factors (presence and degree of multi-disciplinary approach to care, presence of administrative inconveniences) and disease factors (presence of diabetic sensory neuropathy, complexity of disease process). Conclusions The extent of proper foot care amongst diabetic patients is influenced by numerous predisposing and precipitating factors. Further studies can look at further development of the described structure as well as quantitatively defining the various components and factors which make up the described system. PMID:28706922

Transcriptome Profiles of Isolated Murine Achilles Tendon Proper- and Peritenon- Derived Progenitor Cells.

PubMed

Mienaltowski, Michael J; Cánovas, Angela; Fates, Valerie A; Hampton, Angela R; Pechanec, Monica Y; Islas-Trejo, Alma; Medrano, Juan F

2018-06-21

Progenitor cells of the tendon proper and peritenon have unique properties that could impact their utilization in tendon repair strategies. While a few markers have been found to aid in distinguishing progenitors cells from each region, there is great value in identifying more markers. In this study, we hypothesized that RNAseq could be used to improve our understanding of those markers that define these cell types. Transcriptome profiles were generated for pools of mouse Achilles tendon progenitor cells from both regions and catalogues of potential markers were generated. Moreover, common (e.g., glycoprotein, signaling, and proteinaceous extracellular matrix) and unique (e.g., cartilage development versus angiogenesis and muscle contraction) biological processes and molecular functions were described for progenitors from each region. Real-time quantitative PCR of a subset of genes was used to gain insight into the heterogeneity amongst individual progenitor colonies from each region. Markers like Scx, Mkx, Thbs4, and Wnt10a were consistently able to distinguish tendon proper progenitors from peritenon progenitors; expression variability for other genes suggested greater cell type complexity for potential peritenon progenitor markers. This is the first effort to define Achilles tendon progenitor markers by region. Further efforts to investigate the value of these catalogued markers are required by screening more individual colonies of progenitors for more markers. Findings from this study advance efforts in the discernment of cell type specific markers for tendon proper and peritenon progenitor cells; insight into marker sets could improve tracking and sorting strategies for these cells for future therapeutic strategies. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

Toward Facilitative Mentoring and Catalytic Interventions

ERIC Educational Resources Information Center

Smith, Melissa K.; Lewis, Marilyn

2015-01-01

In TESOL teacher mentoring, giving advice can be conceptualized as a continuum, ranging from directive to facilitative feedback. The goal, over time, is to lead toward the facilitative end of the continuum and specifically to catalytic interventions that encourage self-reflection and autonomous learning. This study begins by examining research on…

A deep proper motion catalog within the Sloan digital sky survey footprint

DOE Office of Scientific and Technical Information (OSTI.GOV)

Munn, Jeffrey A.; Harris, Hugh C.; Tilleman, Trudy M.

2014-12-01

A new proper motion catalog is presented, combining the Sloan Digital Sky Survey (SDSS) with second epoch observations in the r band within a portion of the SDSS imaging footprint. The new observations were obtained with the 90prime camera on the Steward Observatory Bok 90 inch telescope, and the Array Camera on the U.S. Naval Observatory, Flagstaff Station, 1.3 m telescope. The catalog covers 1098 square degrees to r = 22.0, an additional 1521 square degrees to r = 20.9, plus a further 488 square degrees of lesser quality data. Statistical errors in the proper motions range from 5 masmore » year{sup −1} at the bright end to 15 mas year{sup −1} at the faint end, for a typical epoch difference of six years. Systematic errors are estimated to be roughly 1 mas year{sup −1} for the Array Camera data, and as much as 2–4 mas year{sup −1} for the 90prime data (though typically less). The catalog also includes a second epoch of r band photometry.« less

Constraining the Mass of the Local Group through Proper Motion Measurements of Local Group Galaxies

NASA Astrophysics Data System (ADS)

Sohn, S. Tony; van der Marel, R.; Anderson, J.

2012-01-01

The Local Group and its two dominant spiral galaxies have been the benchmark for testing many aspects of cosmological and galaxy formation theories. This includes, e.g., dark halo profiles and shapes, substructure and the "missing satellite" problem, and the minimum mass for galaxy formation. But despite the extensive work in all of these areas, our knowledge of the mass of the Milky Way and M31, and thus the total mass of the Local Group remains one of the most poorly established astronomical parameters (uncertain by a factor of 4). One important reason for this problem is the lack of information in tangential motions of galaxies, which can be only obtained through proper motion measurements. In this study, we introduce our projects for measuring absolute proper motions of (1) the dwarf spheroidal galaxy Leo I, (2) M31, and (3) the 4 dwarf galaxies near the edge of the Local Group (Cetus, Leo A, Tucana, and Sag DIG). Results from these three independent measurements will provide important clues to the mass of the Milky Way, M31, and the Local Group as a whole, respectively. We also present our proper motion measurement technique that uses compact background galaxies as astrometric reference sources.

A Protocol-Based Decision for Choosing a Proper Surgical Treatment Option for Carotid Artery Stenosis.

PubMed

Jang, E-Wook; Chung, Joonho; Seo, Kwon-Duk; Suh, Sang Hyun; Kim, Yong Bae; Lee, Kyung-Yul

2015-06-01

There are two established surgical treatment options for carotid artery stenosis. Carotid endarterectomy (CEA) has been accepted as a gold standard for surgical treatment while carotid artery stenting (CAS) has recently become an alternative option. Each treatment option has advantages and disadvantages for the treatment outcomes. We propose a protocol for selection of a proper surgical treatment option for carotid artery stenosis. A total of 192 published articles on management of carotid artery stenosis were reviewed. Preoperatively considerable factors which had been repeatedly noted in those articles for the risk/benefits of CEA or CAS were selected. According to those factors, a protocol with four categories was established. CEA or CAS is indicated when the patient has a symptomatic stenosis ≥ 50%, or when the patient has an asymptomatic stenosis ≥ 80%. Each treatment option has absolute indications and favorable indications. Each absolute indication is scored with three points, and each favorable indication, one point. Based on the highest scores, a proper treatment option (CEA or CAS) is selected. We have been treating patients according to this protocol and evaluating the outcomes of our protocol-based decision because this protocol might be helpful in assessment of risk/benefit for selection of a proper surgical treatment option in patients with carotid artery stenosis.

Facilitators for practice change in Spanish community pharmacy.

PubMed

Gastelurrutia, Miguel A; Benrimoj, S I Charlie; Castrillon, Carla C; de Amezua, María J Casado; Fernandez-Llimos, Fernando; Faus, Maria J

2009-02-01

To identify and prioritise facilitators for practice change in Spanish community pharmacy. Spanish community pharmacies. Qualitative study. Thirty-three semi-structured interviews were conducted with community pharmacists (n = 15) and pharmacy strategists (n = 18), and the results were examined using the content analysis method. In addition, two nominal groups (seven community pharmacists and seven strategists) were formed to identify and prioritise facilitators. Results of both techniques were then triangulated. Facilitators for practice change. Twelve facilitators were identified and grouped into four domains (D1: Pharmacist; D2: Pharmacy as an organisation; D3: Pharmaceutical profession; D4: Miscellaneous). Facilitators identified in D1 include: the need for more clinical education at both pre- and post-graduate levels; the need for clearer and unequivocal messages from professional leaders about the future of the professional practice; and the need for a change in pharmacists' attitudes. Facilitators in D2 are: the need to change the reimbursement system to accommodate cognitive service delivery as well as dispensing; and the need to change the front office of pharmacies. Facilitators identified in D3 are: the need for the Spanish National Professional Association to take a leadership role in the implementation of cognitive services; the need to reduce administrative workload; and the need for universities to reduce the gap between education and research. Other facilitators identified in this study include: the need to increase patients' demand for cognitive services at pharmacies; the need to improve pharmacist-physician relationships; the need for support from health care authorities; and the need for improved marketing of cognitive services and their benefits to society, including physicians and health care authorities. Twelve facilitators were identified. Strategists considered clinical education and pharmacists' attitude as the most important, and

VizieR Online Data Catalog: VVV high proper motion stars. I. Ks<=13.5 stars (Kurtev+, 2017)

NASA Astrophysics Data System (ADS)

Kurtev, R.; Gromadzki, M.; Beamin, J. C.; Folkes, S. L.; Pena Ramirez, K.; Ivanov, V. D.; Borissova, J.; Villanueva, V.; Minniti, D.; Mendez, R.; Lucas, P. W.; Smith, L. C.; Pinfield, D. J.; Kuhn, M. A.; Jones, H. R. A.; Antonova, A.; Yip, A. K. P.

2017-05-01

A catalogue of 3003 high proper motion stars projected on the area of the Vista Variables en via Lactea NIR survey. The positions of the first and the last VVV epochs as well as the 2MASS positions of the majority of the catalogue objects are given. The catalogue also contains photometrical data for these 3003 nearby stars in ZYJHKs filters. The proper motions are calculated one using the VVV only data and VVV-2MASS positions. (2 data files).

Aurora kinases and protein phosphatase 1 mediate chromosome congression through regulation of CENP-E

PubMed Central

Kim, Yumi; Holland, Andrew J.; Lan, Weijie; Cleveland, Don W.

2010-01-01

Summary Opposing roles of Aurora kinases and protein phosphatase 1 (PP1) during mitosis have long been suggested. Here we demonstrate that Aurora kinases A and B phosphorylate a single residue on the kinetochore motor CENP-E. PP1 binds CENP-E via a motif overlapping this phosphorylation site and binding is disrupted by Aurora phosphorylation. Phosphorylation of CENP-E by the Auroras is enriched at spindle poles, disrupting binding of PP1 and reducing CENP-E’s affinity for individual microtubules. This phosphorylation is required for CENP-E-mediated towing of initially polar chromosomes toward the cell center. Kinetochores on such chromosomes cannot make subsequent stable attachment to spindle microtubules when dephosphorylation of CENP-E or rebinding of PP1 to CENP-E is blocked. Thus, an Aurora/PP1 phosphorylation switch modulates CENP-E motor activity as an essential feature of chromosome congression from poles and localized PP1 delivery by CENP-E to the outer kinetochore is necessary for stable microtubule capture by those chromosomes. PMID:20691903

Molecular basis of CENP-C association with the CENP-A nucleosome at yeast centromeres

PubMed Central

Xiao, Hua; Wang, Feng; Wisniewski, Jan; Shaytan, Alexey K.; Ghirlando, Rodolfo; FitzGerald, Peter C.; Huang, Yingzi; Wei, Debbie; Li, Shipeng; Landsman, David; Panchenko, Anna R.; Wu, Carl

2017-01-01

Histone CENP-A-containing nucleosomes play an important role in nucleating kinetochores at centromeres for chromosome segregation. However, the molecular mechanisms by which CENP-A nucleosomes engage with kinetochore proteins are not well understood. Here, we report the finding of a new function for the budding yeast Cse4/CENP-A histone-fold domain interacting with inner kinetochore protein Mif2/CENP-C. Strikingly, we also discovered that AT-rich centromere DNA has an important role for Mif2 recruitment. Mif2 contacts one side of the nucleosome dyad, engaging with both Cse4 residues and AT-rich nucleosomal DNA. Both interactions are directed by a contiguous DNA- and histone-binding domain (DHBD) harboring the conserved CENP-C motif, an AT hook, and RK clusters (clusters enriched for arginine–lysine residues). Human CENP-C has two related DHBDs that bind preferentially to DNA sequences of higher AT content. Our findings suggest that a DNA composition-based mechanism together with residues characteristic for the CENP-A histone variant contribute to the specification of centromere identity. PMID:29074736