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Sample records for features high mitotic

  1. Evaluation of associations between common variation in mitotic regulatory pathways and risk of overall and high grade breast cancer.

    PubMed

    Stevens, Kristen N; Wang, Xianshu; Fredericksen, Zachary; Pankratz, V Shane; Cerhan, James; Vachon, Celine M; Olson, Janet E; Couch, Fergus J

    2011-09-01

    Mitotic regulatory pathways insure proper timing of mitotic entry, sister chromatid cohesion and separation, and cytokinesis. Disruption of this process results in inappropriate chromosome segregation and aneuploidy, and appears to contribute to cancer. Specifically, disregulation and somatic mutation of mitotic regulators has been observed in human cancers, and overexpression of mitotic regulators is common in aggressive and late stage tumors. However, the role of germline variation in mitotic pathways and risk of cancer is not well understood. We tested 1,084 haplotype-tagging and functional variants from 164 genes in mitotic regulatory pathways in 791 Caucasian women with breast cancer and 843 healthy controls for association with risk of overall and high grade breast cancer. Sixty-one single nucleotide polymorphisms (SNPs) from 40 genes were associated (P<0.05) with risk of breast cancer in a log-additive model. In addition, 60 SNPs were associated (P<0.05) with risk of high grade breast cancer. However, none of these associations were significant after Bonferroni correction for multiple testing. In gene-level analyses, CDC25C, SCC1/RAD21, TLK2, and SMC6L1 were associated (P<0.05) with overall breast cancer risk, CDC6, CDC27, SUMO3, RASSF1, KIF2, and CDC14A were associated with high grade breast cancer risk, and EIF3S10 and CDC25A were associated with both. Further investigation in breast and other cancers are needed to understand the influence of inherited variation in mitotic genes on tumor grade and cancer risk. PMID:21607584

  2. High-frequency ultrasound analysis of post-mitotic arrest cell death

    PubMed Central

    Pasternak, Maurice M.; Wirtzfeld, Lauren A.; Kolios, Michael C.; Czarnota, Gregory J.

    2016-01-01

    Non-invasive monitoring of cancer cell death would permit rapid feedback on treatment response. One technique showing such promise is quantitative ultrasound. High-frequency ultrasound spectral radiofrequency analysis was used to study cell death in breast cancer cell samples. Quantitative ultrasound parameters, including attenuation, spectral slope, spectral 0-MHz-intercept, midband fit, and fitted parameters displayed significant changes with paclitaxel-induced cell death, corresponding to observations of morphological changes seen in histology and electron microscopy. In particular, a decrease in spectral slope from 0.24±0.07 dB/MHz to 0.04±0.09 dB/MHz occurred over 24 hours of treatment time and was identified as an ultrasound parameter capable of differentiating post-mitotic arrest cell death from classical apoptosis. The formation of condensed chromatin aggregates of 1 micron or greater in size increased the number of intracellular scatterers, consistent with a hypothesis that nuclear material is a primary source of ultrasound scattering in dying cells. It was demonstrated that the midband fit quantitatively correlated to cell death index, with a Pearson R-squared value of 0.99 at p<0.01. These results suggest that high-frequency ultrasound can not only qualitatively assess the degree of cancer cell death, but may be used to quantify the efficacy of chemotherapeutic treatments. PMID:27226984

  3. High throughput screening of natural products for anti-mitotic effects in MDA-MB-231 human breast carcinoma cells.

    PubMed

    Mazzio, E; Badisa, R; Mack, N; Deiab, S; Soliman, K F A

    2014-06-01

    Some of the most effective anti-mitotic microtubule-binding agents, such as paclitaxel (Taxus brevifolia) were originally discovered through robust National Cancer Institute botanical screenings. In this study, a high-through put microarray format was utilized to screen 897 aqueous extracts of commonly used natural products (0.00015-0.5 mg/mL) relative to paclitaxel for anti-mitotic effects (independent of toxicity) on proliferation of MDA-MB-231 cells. The data obtained showed that less than 1.34 % of the extracts tested showed inhibitory growth (IG50 ) properties <0.0183 mg/mL. The most potent anti-mitotics (independent of toxicity) were Mandrake root (Podophyllum peltatum), Truja twigs (Thuja occidentalis), Colorado desert mistletoe (Phoradendron flavescens), Tou Gu Cao [symbol: see text] Speranskia herb (Speranskia tuberculata), Bentonite clay, Bunge root (Pulsatilla chinensis), Brucea fruit (Brucea javanica), Madder root (Rubia tinctorum), Gallnut of Chinese Sumac (Melaphis chinensis), Elecampane root (Inula Helenium), Yuan Zhi [symbol: see text] root (Polygala tenuifolia), Pagoda Tree fruit (Melia Toosendan), Stone root (Collinsonia Canadensis), and others such as American Witchhazel, Arjun, and Bladderwrack. The strongest tumoricidal herbs identified from amongst the subset evaluated for anti-mitotic properties were wild yam (Dioscorea villosa), beth root (Trillium Pendulum), and alkanet root (Lithospermum canescens). Additional data was obtained on a lesser-recognized herb: (S. tuberculata), which showed growth inhibition on BT-474 (human ductal breast carcinoma) and Ishikawa (human endometrial adenocarcinoma) cells with ability to block replicative DNA synthesis, leading to G2 arrest in MDA-MB-231 cells. In conclusion, these findings present relative potency of anti-mitotic natural plants that are effective against human breast carcinoma MDA-MB-231 cell division. PMID:24105850

  4. High throughput screening of natural products for anti-mitotic effects in MDA-MB-231 human breast carcinoma cells

    PubMed Central

    Mazzio, E; Badisa, R; Mack, N; Deiab, S; Soliman, KFA

    2013-01-01

    Some of the most effective anti-mitotic microtubule-binding agents, such as paclitaxel (Taxus brevifolia) were originally discovered through robust NCI botanical screenings. In this study, a high-through microarray format was utilized to screen 897 aqueous extracts of commonly used natural products (0.00015–0.5 mg/ml) relative to paclitaxel for anti-mitotic effects (independent of toxicity) on proliferation of MDA-MB-231 cells. The data obtained showed that less than 1.34 % tested showed inhibitory growth (IG50) properties <0.0183 mg/ml. The most potent anti-mitotics (independent of toxicity) were Mandrake root (Podophyllum peltatum), Truja Twigs (Thuja occidentalis), Colorado desert mistletoe (Phoradendron flavescens), Tou Gu Cao Speranskia Herb (Speranskia tuberculata), Bentonite Clay, Bunge Root (Pulsatilla chinensis), Brucea Fruit (Brucea javanica), Madder Root (Rubia tinctorum), Gallnut of Chinese Sumac (Melaphis chinensis), Elecampane Root (Inula Helenium), Yuan Zhi Root (Polygala tenuifolia), Pagoda Tree Fruit (Melia Toosendan), Stone Root (Collinsonia Canadensis) and others such as American Witchhazel, Arjun and Bladderwrack. The strongest tumoricidal herbs identified from amongst the subset evaluated for anti-mitotic properties were wild yam (Dioscorea villosa), beth-root (Trillium Pendulum) and alkanet-root (Lithospermum canescens). Additional data was obtained on a lesser-recognized herb: (Speranskia tuberculata) which showed growth inhibition on BT-474 (human ductal breast carcinoma) and Ishikawa (human endometrial adenocarcinoma) cells with ability to block replicative DNA synthesis leading to G2 arrest in MDA-MB-231 cells. In conclusion, these findings present relative potency of natural anti-mitotic resources effective against human breast carcinoma MDA-MB-231 cell division. PMID:24105850

  5. Nup2 requires a highly divergent partner, NupA, to fulfill functions at nuclear pore complexes and the mitotic chromatin region

    PubMed Central

    Markossian, Sarine; Suresh, Subbulakshmi; Osmani, Aysha H.; Osmani, Stephen A.

    2015-01-01

    Chromatin and nuclear pore complexes (NPCs) undergo dramatic changes during mitosis, which in vertebrates and Aspergillus nidulans involves movement of Nup2 from NPCs to the chromatin region to fulfill unknown functions. This transition is shown to require the Cdk1 mitotic kinase and be promoted prematurely by ectopic expression of the NIMA kinase. Nup2 localizes with a copurifying partner termed NupA, a highly divergent yet essential NPC protein. NupA and Nup2 locate throughout the chromatin region during prophase but during anaphase move to surround segregating DNA. NupA function is shown to involve targeting Nup2 to its interphase and mitotic locations. Deletion of either Nup2 or NupA causes identical mitotic defects that initiate a spindle assembly checkpoint (SAC)–dependent mitotic delay and also cause defects in karyokinesis. These mitotic problems are not caused by overall defects in mitotic NPC disassembly–reassembly or general nuclear import. However, without Nup2 or NupA, although the SAC protein Mad1 locates to its mitotic locations, it fails to locate to NPCs normally in G1 after mitosis. Collectively the study provides new insight into the roles of Nup2 and NupA during mitosis and in a surveillance mechanism that regulates nucleokinesis when mitotic defects occur after SAC fulfillment. PMID:25540430

  6. The Drosophila Salivary Gland Chromocenter Contains Highly Polytenized Subdomains of Mitotic Heterochromatin

    PubMed Central

    Zhang, P.; Spradling, A. C.

    1995-01-01

    Peri-centromeric regions of Drosophila melanogaster chromosomes appear heterochromatic in mitotic cells and become greatly underrepresented in giant polytene chromosomes, where they aggregate into a central mass called the chromocenter. We used P elements inserted at sites dispersed throughout much of the mitotic heterochromatin to analyze the fate of 31 individual sites during polytenization. Analysis of DNA sequences flanking many of these elements revealed that middle repetitive or unique sequence DNAs frequently are interspersed with satellite DNAs in mitotic heterochromatin. All nine Y chromosome sites tested were underrepresented >20-fold on Southern blots of polytene DNA and were rarely or never detected by in situ hybridization to salivary gland chromosomes. In contrast, nine tested insertions in autosomal centromeric heterochromatin were represented fully in salivary gland DNA, despite the fact that at least six were located proximal to known blocks of satellite DNA. The inserted sequences formed diverse, site-specific morphologies in the chromocenter of salivary gland chromosomes, suggesting that domains dispersed at multiple sites in the centromeric heterochromatin of mitotic chromosomes contribute to polytene β-heterochromatin. We suggest that regions containing heterochromatic genes are organized into dispersed chromatin configurations that are important for their function in vivo. PMID:7713423

  7. Extremely high-dimensional feature selection via feature generating samplings.

    PubMed

    Li, Shutao; Wei, Dan

    2014-06-01

    To select informative features on extremely high-dimensional problems, in this paper, a sampling scheme is proposed to enhance the efficiency of recently developed feature generating machines (FGMs). Note that in FGMs O(mlogr) time complexity should be taken to order the features by their scores; the entire computational cost of feature ordering will become unbearable when m is very large, for example, m > 10(11) , where m is the feature dimensionality and r is the size of the selected feature subset. To solve this problem, in this paper, we propose a feature generating sampling method, which can reduce this computational complexity to O(Gslog(G)+G(G+log(G))) while preserving the most informative features in a feature buffer, where Gs is the maximum number of nonzero features for each instance and G is the buffer size. Moreover, we show that our proposed sampling scheme can be deemed as the birth-death process based on random processes theory, which guarantees to include most of the informative features for feature selections. Empirical studies on real-world datasets show the effectiveness of the proposed sampling method. PMID:23864272

  8. Leiomyosarcoma arising in a patient with prior mitotically active leiomyoma.

    PubMed

    Kim, Jin Hwi; Choi, Young Jin; Kim, Dong Chul; Lee, Sung Jong

    2010-02-01

    Mitotically active leiomyomas exhibiting insignificant cytologic atypia (none to mild) with an increased mitotic count of 5-20 mitotic figures per 10 high power fields, and without coagulative tumor cell necrosis or atypical mitoses, tend to have a benign clinical course. We encountered a case of a mitotically active leiomyoma and malignant transformation after a total hysterectomy. The recurrent multiple abdominal masses were removed surgically; most of them were benign leiomyomas, but some were leiomyosarcomas. Although most mitotically active leiomyomas have a benign clinical course, these lesions can recur and have the potential for malignant transformation. Therefore, patients with mitotically active leiomyomas require close follow-up. PMID:20178549

  9. Building mitotic chromosomes

    PubMed Central

    Ohta, Shinya; Wood, Laura; Bukowski-Wills, Jimi-Carlo; Rappsilber, Juri; Earnshaw, William C

    2011-01-01

    Mitotic chromosomes are the iconic structures into which the genome is packaged to ensure its accurate segregation during mitosis. Although they have appeared on countless journal cover illustrations, there remains no consensus on how the chromatin fiber is packaged during mitosis. In fact, work in recent years has both added to existing controversies and sparked new ones. By contrast, there has been very significant progress in determining the protein composition of isolated mitotic chromosomes. Here, we discuss recent studies of chromosome organization and provide an in depth description of the latest proteomics studies, which have at last provided us with a definitive proteome for vertebrate chromosomes. PMID:20974528

  10. Building mitotic chromosomes.

    PubMed

    Ohta, Shinya; Wood, Laura; Bukowski-Wills, Jimi-Carlo; Rappsilber, Juri; Earnshaw, William C

    2011-02-01

    Mitotic chromosomes are the iconic structures into which the genome is packaged to ensure its accurate segregation during mitosis. Although they have appeared on countless journal cover illustrations, there remains no consensus on how the chromatin fiber is packaged during mitosis. In fact, work in recent years has both added to existing controversies and sparked new ones. By contrast, there has been very significant progress in determining the protein composition of isolated mitotic chromosomes. Here, we discuss recent studies of chromosome organization and provide an in depth description of the latest proteomics studies, which have at last provided us with a definitive proteome for vertebrate chromosomes. PMID:20974528

  11. A High Throughput, Whole Cell Screen for Small Molecule Inhibitors of the Mitotic Spindle Checkpoint Identifies OM137, a Novel Aurora Kinase Inhibitor

    PubMed Central

    DeMoe, Joanna H.; Santaguida, Stefano; Daum, John R.; Musacchio, Andrea; Gorbsky, Gary J.

    2008-01-01

    In mitosis the kinetochores of chromosomes that lack full microtubule attachments and/or mechanical tension activate a signaling pathway called the mitotic spindle checkpoint that blocks progression into anaphase and prevents premature segregation of the chromatids until chromosomes become aligned at the metaphase plate (1). The spindle checkpoint is responsible for arresting cells in mitosis in response to chemotherapeutic spindle poisons such as paclitaxel or vinblastine. Some cancer cells show a weakened checkpoint signaling system that may contribute to chromosome instability in tumors. Since complete absence of the spindle checkpoint leads to catastrophic cell division, we reasoned that drugs targeting the checkpoint might provide a therapeutic window in which the checkpoint would be eliminated in cancer cells but sufficiently preserved in normal cells. We developed an assay to identify lead compounds that inhibit the spindle checkpoint. Most cells respond to microtubule drugs by activating the spindle checkpoint and arresting in mitosis with a rounded morphology. Our assay depended on the ability of checkpoint inhibitor compounds to drive mitotic exit and cause cells to flatten onto the substrate in the continuous presence of microtubule drugs. In this study we characterize one of the compounds, OM137, as an inhibitor of Aurora kinases. We find that this compound is growth inhibitory to cultured cells when applied at high concentration and potentiates the growth inhibitory effects of subnanomolar concentrations of paclitaxel. PMID:19190331

  12. Evaluation of Tumor Cell Proliferation by Ki-67 Expression and Mitotic Count in Lymph Node Metastases from Breast Cancer

    PubMed Central

    Aziz, Sura; Wik, Elisabeth; Davidsen, Benedicte; Aas, Hans; Aas, Turid; Akslen, Lars A.

    2016-01-01

    Few studies have addressed the risk of recurrence by assessing proliferation markers in lymph node metastasis from breast cancer. Here, we aimed to examine Ki-67 expression and mitotic count in lymph nodes in comparison with primary tumors. A cohort of node positive breast cancer (n = 168) was studied as a part of the prospective Norwegian Breast Cancer Screening Program (1996–2009). The percentage of Ki-67 positivity was counted per 500 tumor cells in hot-spot areas (x630). Mitotic count was conducted in the most cellular and mitotic active areas in 10 high power fields (x400). Our results showed that Ki-67 and mitotic count were significantly correlated between primary tumor and lymph nodes (Spearman`s correlation 0. 56 and 0.46, respectively) and were associated with most of the histologic features of the primary tumor. Univariate survival analysis (log-rank test) showed that high Ki-67 and mitotic count in the primary tumor and lymph node metastasis significantly predicted risk of recurrence. In multivariate analysis, mitotic count in the lymph node metastasis was an independent predictor of tumor recurrence. In conclusion, proliferation markers in lymph node metastases significantly predicted disease free survival in node positive breast cancer. PMID:26954367

  13. Highly Nonlinear Features of Electron Diffusion Region

    NASA Astrophysics Data System (ADS)

    Singh, N.

    2007-12-01

    Using a fully three-dimensional Particle-in-cell code and real ion to electron mass ratio of 1836 in a hydrogen plasma, we simulated electrodynamics in an extremely thin current sheet (CS) with and without a guide field. Simulations reveal several highly nonlinear features of electron diffusion region as measured from satellites in magneto-tail and magnetopause regions. These features include: (i) ion acceleration by the Hall electric field like in a ion thruster, (ii) bifurcated current sheet by current disruption (iii) fine-scale step-like structure in the CS magnetic field profile, (iv) strong clumping of electrons and ions in the midst of the CS, (v) spiky perpendicular electric fields confined within the CS, (vi) generation of current layers parallel to the reconnecting magnetic field in the presence of a guide field, (vii) strong modification in the spatial distribution of the guide field across the CS, and (viii) electron acceleration to relativistic energies by the electromagnetic turbulence. Results from the satellite observations and simulations are compared quantitatively.

  14. Mitotic chromosome condensation in vertebrates

    SciTech Connect

    Vagnarelli, Paola

    2012-07-15

    Work from several laboratories over the past 10-15 years has revealed that, within the interphase nucleus, chromosomes are organized into spatially distinct territories [T. Cremer, C. Cremer, Chromosome territories, nuclear architecture and gene regulation in mammalian cells, Nat. Rev. Genet. 2 (2001) 292-301 and T. Cremer, M. Cremer, S. Dietzel, S. Muller, I. Solovei, S. Fakan, Chromosome territories-a functional nuclear landscape, Curr. Opin. Cell Biol. 18 (2006) 307-316]. The overall compaction level and intranuclear location varies as a function of gene density for both entire chromosomes [J.A. Croft, J.M. Bridger, S. Boyle, P. Perry, P. Teague,W.A. Bickmore, Differences in the localization and morphology of chromosomes in the human nucleus, J. Cell Biol. 145 (1999) 1119-1131] and specific chromosomal regions [N.L. Mahy, P.E. Perry, S. Gilchrist, R.A. Baldock, W.A. Bickmore, Spatial organization of active and inactive genes and noncoding DNA within chromosome territories, J. Cell Biol. 157 (2002) 579-589] (Fig. 1A, A'). In prophase, when cyclin B activity reaches a high threshold, chromosome condensation occurs followed by Nuclear Envelope Breakdown (NEB) [1]. At this point vertebrate chromosomes appear as compact structures harboring an attachment point for the spindle microtubules physically recognizable as a primary constriction where the two sister chromatids are held together. The transition from an unshaped interphase chromosome to the highly structured mitotic chromosome (compare Figs. 1A and B) has fascinated researchers for several decades now; however a definite picture of how this process is achieved and regulated is not yet in our hands and it will require more investigation to comprehend the complete process. From a biochemical point of view a vertebrate mitotic chromosomes is composed of DNA, histone proteins (60%) and non-histone proteins (40%) [6]. I will discuss below what is known to date on the contribution of these two different classes of proteins and their co-operation in establishing the final mitotic chromosome structure.

  15. The Conserved N-Terminal Region of the Mitotic Checkpoint Protein BUBR1: A Putative TPR Motif of High Surface Activity

    PubMed Central

    Bolanos-Garcia, V. M.; Beaufils, S.; Renault, A.; Grossmann, J. G.; Brewerton, S.; Lee, M.; Venkitaraman, A.; Blundell, T. L.

    2005-01-01

    BUBR1, a key component of the mitotic spindle checkpoint, is a multidomain protein kinase that is activated in response to kinetochore tension. Although BUB1 and BUBR1 play an important role in cell division, very little is known about their structural characteristics. We show that the conserved N-terminal region of BUBR1, comprising residues 1–204, is a globular domain of high α-helical content (≈60%), stable in the pH range 4–9 and probably organized as a tetratricopeptide motif repeat (TPR), most closely resembling residues 16–181 of protein phosphatase 5. Because the latter presents a continuous amphipathic groove and is regulated by binding certain fatty acids, we compared the properties of BUBR1(1–204) and TPR-PP5(16–181) at air/water interfaces and found that both proteins exhibited a similar surface activity and formed stable, rigid monolayers. The deletion of a region that probably comprises several α-helices of BUBR1 indicates that long-range interactions are essential for the stability of the N-terminal domain. The presence of the putative TPR motif strongly suggests that the N-terminal domain of BUBR1 is involved in direct protein-protein interactions and/or protein-lipid interactions. PMID:16040755

  16. Chemically diverse microtubule stabilizing agents initiate distinct mitotic defects and dysregulated expression of key mitotic kinases.

    PubMed

    Rohena, Cristina C; Peng, Jiangnan; Johnson, Tyler A; Crews, Phillip; Mooberry, Susan L

    2013-04-15

    Microtubule stabilizers are some of the most successful drugs used in the treatment of adult solid tumors and yet the molecular events responsible for their antimitotic actions are not well defined. The mitotic events initiated by three structurally and biologically diverse microtubule stabilizers; taccalonolide AJ, laulimalide/fijianolide B and paclitaxel were studied. These microtubule stabilizers cause the formation of aberrant, but structurally distinct mitotic spindles leading to the hypothesis that they differentially affect mitotic signaling. Each microtubule stabilizer initiated different patterns of expression of key mitotic signaling proteins. Taccalonolide AJ causes centrosome separation and disjunction failure to a much greater extent than paclitaxel or laulimalide, which is consistent with the distinct defects in expression and activation of Plk1 and Eg5 caused by each stabilizer. Localization studies revealed that TPX2 and Aurora A are associated with each spindle aster formed by each stabilizer. This suggests a common mechanism of aster formation. However, taccalonolide AJ also causes pericentrin accumulation on every spindle aster. The presence of pericentrin at every spindle aster initiated by taccalonolide AJ might facilitate the maintenance and stability of the highly focused asters formed by this stabilizer. Laulimalide and paclitaxel cause completely different patterns of expression and activation of these proteins, as well as phenotypically different spindle phenotypes. Delineating how diverse microtubule stabilizers affect mitotic signaling pathways could identify key proteins involved in modulating sensitivity and resistance to the antimitotic actions of these compounds. PMID:23399639

  17. Mitotic Spindle Form and Function

    PubMed Central

    Winey, Mark; Bloom, Kerry

    2012-01-01

    The Saccharomyces cerevisiae mitotic spindle in budding yeast is exemplified by its simplicity and elegance. Microtubules are nucleated from a crystalline array of proteins organized in the nuclear envelope, known as the spindle pole body in yeast (analogous to the centrosome in larger eukaryotes). The spindle has two classes of nuclear microtubules: kinetochore microtubules and interpolar microtubules. One kinetochore microtubule attaches to a single centromere on each chromosome, while approximately four interpolar microtubules emanate from each pole and interdigitate with interpolar microtubules from the opposite spindle to provide stability to the bipolar spindle. On the cytoplasmic face, two to three microtubules extend from the spindle pole toward the cell cortex. Processes requiring microtubule function are limited to spindles in mitosis and to spindle orientation and nuclear positioning in the cytoplasm. Microtubule function is regulated in large part via products of the 6 kinesin gene family and the 1 cytoplasmic dynein gene. A single bipolar kinesin (Cin8, class Kin-5), together with a depolymerase (Kip3, class Kin-8) or minus-end-directed kinesin (Kar3, class Kin-14), can support spindle function and cell viability. The remarkable feature of yeast cells is that they can survive with microtubules and genes for just two motor proteins, thus providing an unparalleled system to dissect microtubule and motor function within the spindle machine. PMID:22491889

  18. Isolation of Human Mitotic Protein Phosphatase Complexes: Identification of a Complex between Protein Phosphatase 1 and the RNA Helicase Ddx21

    PubMed Central

    De Wever, Veerle; Lloyd, David C.; Nasa, Isha; Nimick, Mhairi; Trinkle-Mulcahy, Laura; Gourlay, Robert; Morrice, Nick; Moorhead, Greg B. G.

    2012-01-01

    Metazoan mitosis requires remodelling of sub-cellular structures to ensure proper division of cellular and genetic material. Faults often lead to genomic instability, cell cycle arrests and disease onset. These key structural changes are under tight spatial-temporal and post-translational control, with crucial roles for reversible protein phosphorylation. The phosphoprotein phosphatases PP1 and PP2A are paramount for the timely execution of mitotic entry and exit but their interaction partners and substrates are still largely unresolved. High throughput, mass-spectrometry based studies have limited sensitivity for the detection of low-abundance and transient complexes, a typical feature of many protein phosphatase complexes. Moreover, the limited timeframe during which mitosis takes place reduces the likelihood of identifying mitotic phosphatase complexes in asynchronous cells. Hence, numerous mitotic protein phosphatase complexes still await identification. Here we present a strategy to enrich and identify serine/threonine protein phosphatase complexes at the mitotic spindle. We thus identified a nucleolar RNA helicase, Ddx21/Gu, as a novel, direct PP1 interactor. Furthermore, our results place PP1 within the toposome, a Topoisomerase II alpha (TOPOIIα) containing complex with a key role in mitotic chromatin regulation and cell cycle progression, possibly via regulated protein phosphorylation. This study provides a strategy for the identification of further mitotic PP1 partners and the unravelling of PP1 functions during mitosis. PMID:22761809

  19. ControlNet features high speed

    SciTech Connect

    McEldowney, D.

    1996-11-01

    ControlNet is a high-speed, high-capacity network providing a connection among controllers and I/O subsystems. It was designed for applications in which data integrity, determinism, high speeds, and high data capacities are required. ControlNet addresses applications needing tighter control over processes as well as demanding remote I/O or interlocked PLC applications, both discrete- and process-related. Some examples include high-speed conveyors, transfer lines, cut-to-length lines, high-speed assembly, bottling, and packaging. Process examples, or those typically requiring heavy remote analog I/O, include water/wastewater, test stands, chemical, beverage, food, marine control, and utility balance-of-plant.

  20. Mitotic Exit Control as an Evolved Complex System

    SciTech Connect

    Bosl, W; Li, R

    2005-04-25

    The exit from mitosis is the last critical decision a cell has to make during a division cycle. A complex regulatory system has evolved to evaluate the success of mitotic events and control this decision. Whereas outstanding genetic work in yeast has led to rapid discovery of a large number of interacting genes involved in the control of mitotic exit, it has also become increasingly difficult to comprehend the logic and mechanistic features embedded in the complex molecular network. Our view is that this difficulty stems in part from the attempt to explain mitotic exit control using concepts from traditional top-down engineering design, and that exciting new results from evolutionary engineering design applied to networks and electronic circuits may lend better insights. We focus on four particularly intriguing features of the mitotic exit control system: the two-stepped release of Cdc14; the self-activating nature of Tem1 GTPase; the spatial sensor associated with the spindle pole body; and the extensive redundancy in the mitotic exit network. We attempt to examine these design features from the perspective of evolutionary design and complex system engineering.

  1. Energy Conservation Featured in Illinois High School

    ERIC Educational Resources Information Center

    Modern Schools, 1976

    1976-01-01

    The William Fremd High School in Palatine, Illinois, scheduled to open in 1977, is being built with energy conservation uppermost in mind. In this system, 70 heat pumps will heat and cool 300,000 square feet of educational facilities. (Author/MLF)

  2. High dimensional feature reduction via projection pursuit

    NASA Technical Reports Server (NTRS)

    Jimenez, Luis; Landgrebe, David

    1994-01-01

    The recent development of more sophisticated remote sensing systems enables the measurement of radiation in many more spectral intervals than previously possible. An example of that technology is the AVIRIS system, which collects image data in 220 bands. As a result of this, new algorithms must be developed in order to analyze the more complex data effectively. Data in a high dimensional space presents a substantial challenge, since intuitive concepts valid in a 2-3 dimensional space to not necessarily apply in higher dimensional spaces. For example, high dimensional space is mostly empty. This results from the concentration of data in the corners of hypercubes. Other examples may be cited. Such observations suggest the need to project data to a subspace of a much lower dimension on a problem specific basis in such a manner that information is not lost. Projection Pursuit is a technique that will accomplish such a goal. Since it processes data in lower dimensions, it should avoid many of the difficulties of high dimensional spaces. In this paper, we begin the investigation of some of the properties of Projection Pursuit for this purpose.

  3. Micromechanics of human mitotic chromosomes

    NASA Astrophysics Data System (ADS)

    Sun, Mingxuan; Kawamura, Ryo; Marko, John F.

    2011-02-01

    Eukaryote cells dramatically reorganize their long chromosomal DNAs to facilitate their physical segregation during mitosis. The internal organization of folded mitotic chromosomes remains a basic mystery of cell biology; its understanding would likely shed light on how chromosomes are separated from one another as well as into chromosome structure between cell divisions. We report biophysical experiments on single mitotic chromosomes from human cells, where we combine micromanipulation, nano-Newton-scale force measurement and biochemical treatments to study chromosome connectivity and topology. Results are in accord with previous experiments on amphibian chromosomes and support the 'chromatin network' model of mitotic chromosome structure. Prospects for studies of chromosome-organizing proteins using siRNA expression knockdowns, as well as for differential studies of chromosomes with and without mutations associated with genetic diseases, are also discussed.

  4. Forced mitotic entry of S-phase cells as a therapeutic strategy induced by inhibition of WEE1.

    PubMed

    Aarts, Marieke; Sharpe, Rachel; Garcia-Murillas, Isaac; Gevensleben, Heidrun; Hurd, Melissa S; Shumway, Stuart D; Toniatti, Carlo; Ashworth, Alan; Turner, Nicholas C

    2012-06-01

    Inhibition of the protein kinase WEE1 synergizes with chemotherapy in preclinical models and WEE1 inhibitors are being explored as potential cancer therapies. Here, we investigate the mechanism that underlies this synergy. We show that WEE1 inhibition forces S-phase-arrested cells directly into mitosis without completing DNA synthesis, resulting in highly abnormal mitoses characterized by dispersed chromosomes and disorganized bipolar spindles, ultimately resulting in mitotic exit with gross micronuclei formation and apoptosis. This mechanism of cell death is shared by CHK1 inhibitors, and combined WEE1 and CHK1 inhibition forces mitotic entry from S-phase in the absence of chemotherapy. We show that p53/p21 inactivation combined with high expression of mitotic cyclins and EZH2 predispose to mitotic entry during S-phase with cells reliant on WEE1 to prevent premature cyclin-dependent kinase (CDK)1 activation. These features are characteristic of aggressive breast, and other, cancers for which WEE1 inhibitor combinations represent a promising targeted therapy. PMID:22628408

  5. High-resolution Urban Image Classification Using Extended Features

    SciTech Connect

    Vatsavai, Raju

    2011-01-01

    High-resolution image classification poses several challenges because the typical object size is much larger than the pixel resolution. Any given pixel (spectral features at that location) by itself is not a good indicator of the object it belongs to without looking at the broader spatial footprint. Therefore most modern machine learning approaches that are based on per-pixel spectral features are not very effective in high- resolution urban image classification. One way to overcome this problem is to extract features that exploit spatial contextual information. In this study, we evaluated several features in- cluding edge density, texture, and morphology. Several machine learning schemes were tested on the features extracted from a very high-resolution remote sensing image and results were presented.

  6. Unsupervised Feature Learning for High-Resolution Satellite Image Classification

    SciTech Connect

    Cheriyadat, Anil M

    2013-01-01

    The rich data provided by high-resolution satellite imagery allow us to directly model geospatial neighborhoods by understanding their spatial and structural patterns. In this paper we explore an unsupervised feature learning approach to model geospatial neighborhoods for classification purposes. While pixel and object based classification approaches are widely used for satellite image analysis, often these approaches exploit the high-fidelity image data in a limited way. In this paper we extract low-level features to characterize the local neighborhood patterns. We exploit the unlabeled feature measurements in a novel way to learn a set of basis functions to derive new features. The derived sparse feature representation obtained by encoding the measured features in terms of the learned basis function set yields superior classification performance. We applied our technique on two challenging image datasets: ORNL dataset representing one-meter spatial resolution satellite imagery representing five land-use categories and, UCMERCED dataset consisting of 21 different categories representing sub-meter resolution overhead imagery. Our results are highly promising and, in the case of UCMERCED dataset we outperform the best results obtained for this dataset. We show that our feature extraction and learning methods are highly effective in developing a detection system that can be used to automatically scan large-scale high-resolution satellite imagery for detecting large-facility.

  7. High Dimensional Classification Using Features Annealed Independence Rules.

    PubMed

    Fan, Jianqing; Fan, Yingying

    2008-01-01

    Classification using high-dimensional features arises frequently in many contemporary statistical studies such as tumor classification using microarray or other high-throughput data. The impact of dimensionality on classifications is largely poorly understood. In a seminal paper, Bickel and Levina (2004) show that the Fisher discriminant performs poorly due to diverging spectra and they propose to use the independence rule to overcome the problem. We first demonstrate that even for the independence classification rule, classification using all the features can be as bad as the random guessing due to noise accumulation in estimating population centroids in high-dimensional feature space. In fact, we demonstrate further that almost all linear discriminants can perform as bad as the random guessing. Thus, it is paramountly important to select a subset of important features for high-dimensional classification, resulting in Features Annealed Independence Rules (FAIR). The conditions under which all the important features can be selected by the two-sample t-statistic are established. The choice of the optimal number of features, or equivalently, the threshold value of the test statistics are proposed based on an upper bound of the classification error. Simulation studies and real data analysis support our theoretical results and demonstrate convincingly the advantage of our new classification procedure. PMID:19169416

  8. Adenovirus Replaces Mitotic Checkpoint Controls

    PubMed Central

    Turner, Roberta L.; Groitl, Peter; Dobner, Thomas

    2015-01-01

    ABSTRACT Infection with adenovirus triggers the cellular DNA damage response, elements of which include cell death and cell cycle arrest. Early adenoviral proteins, including the E1B-55K and E4orf3 proteins, inhibit signaling in response to DNA damage. A fraction of cells infected with an adenovirus mutant unable to express the E1B-55K and E4orf3 genes appeared to arrest in a mitotic-like state. Cells infected early in G1 of the cell cycle were predisposed to arrest in this state at late times of infection. This arrested state, which displays hallmarks of mitotic catastrophe, was prevented by expression of either the E1B-55K or the E4orf3 genes. However, E1B-55K mutant virus-infected cells became trapped in a mitotic-like state in the presence of the microtubule poison colcemid, suggesting that the two viral proteins restrict entry into mitosis or facilitate exit from mitosis in order to prevent infected cells from arresting in mitosis. The E1B-55K protein appeared to prevent inappropriate entry into mitosis through its interaction with the cellular tumor suppressor protein p53. The E4orf3 protein facilitated exit from mitosis by possibly mislocalizing and functionally inactivating cyclin B1. When expressed in noninfected cells, E4orf3 overcame the mitotic arrest caused by the degradation-resistant R42A cyclin B1 variant. IMPORTANCE Cells that are infected with adenovirus type 5 early in G1 of the cell cycle are predisposed to arrest in a mitotic-like state in a p53-dependent manner. The adenoviral E1B-55K protein prevents entry into mitosis. This newly described activity for the E1B-55K protein appears to depend on the interaction between the E1B-55K protein and the tumor suppressor p53. The adenoviral E4orf3 protein facilitates exit from mitosis, possibly by altering the intracellular distribution of cyclin B1. By preventing entry into mitosis and by promoting exit from mitosis, these adenoviral proteins act to prevent the infected cell from arresting in a mitotic-like state. PMID:25694601

  9. Mitotic Kinases and p53 Signaling

    PubMed Central

    Ha, Geun-Hyoung; Breuer, Eun-Kyoung Yim

    2012-01-01

    Mitosis is tightly regulated and any errors in this process often lead to aneuploidy, genomic instability, and tumorigenesis. Deregulation of mitotic kinases is significantly associated with improper cell division and aneuploidy. Because of their importance during mitosis and the relevance to cancer, mitotic kinase signaling has been extensively studied over the past few decades and, as a result, several mitotic kinase inhibitors have been developed. Despite promising preclinical results, targeting mitotic kinases for cancer therapy faces numerous challenges, including safety and patient selection issues. Therefore, there is an urgent need to better understand the molecular mechanisms underlying mitotic kinase signaling and its interactive network. Increasing evidence suggests that tumor suppressor p53 functions at the center of the mitotic kinase signaling network. In response to mitotic spindle damage, multiple mitotic kinases phosphorylate p53 to either activate or deactivate p53-mediated signaling. p53 can also regulate the expression and function of mitotic kinases, suggesting the existence of a network of mutual regulation, which can be positive or negative, between mitotic kinases and p53 signaling. Therefore, deciphering this regulatory network will provide knowledge to overcome current limitations of targeting mitotic kinases and further improve the results of targeted therapy. PMID:22852086

  10. Targeting mitotic exit for cancer treatment.

    PubMed

    Wäsch, Ralph

    2011-07-01

    Several chemotherapeutic drugs interfere with assembly of the mitotic spindle of cancer cells, leading to mitotic arrest, mediated by the spindle assembly checkpoint (SAC). However, cancer cells may be SAC-deficient and survive such treatment, due to mitotic slippage. Sensing of defective spindle assembly by the SAC, causes inhibition of the anaphase-promoting complex or cyclosome (APC/C), and blocks mitotic exit, by stabilizing APC/C substrates, such as cyclin B. Mitotic slippage may occur due to residual APC/C activity and slow cyclin B degradation. Recent preclinical data suggests that targeting mitotic exit by blocking APC/C activity is a much more efficient therapeutic approach than disturbing mitotic spindle assembly. PMID:21476883

  11. Exploring KM Features of High-Performance Companies

    NASA Astrophysics Data System (ADS)

    Wu, Wei-Wen

    2007-12-01

    For reacting to an increasingly rival business environment, many companies emphasize the importance of knowledge management (KM). It is a favorable way to explore and learn KM features of high-performance companies. However, finding out the critical KM features of high-performance companies is a qualitative analysis problem. To handle this kind of problem, the rough set approach is suitable because it is based on data-mining techniques to discover knowledge without rigorous statistical assumptions. Thus, this paper explored KM features of high-performance companies by using the rough set approach. The results show that high-performance companies stress the importance on both tacit and explicit knowledge, and consider that incentives and evaluations are the essentials to implementing KM.

  12. The Zebra fish cassiopeia Mutant Reveals that SIL Is Required for Mitotic Spindle Organization▿ §

    PubMed Central

    Pfaff, Kathleen L.; Straub, Christian T.; Chiang, Ken; Bear, Daniel M.; Zhou, Yi; Zon, Leonard I.

    2007-01-01

    A critical step in cell division is formation of the mitotic spindle, which is a bipolar array of microtubules that mediates chromosome separation. Here, we report that the SCL-interrupting locus (SIL), a vertebrate-specific cytosolic protein, is necessary for proper mitotic spindle organization in zebrafish and human cells. A homozygous lethal zebrafish mutant, cassiopeia (csp), was identified by a genetic screen for mitotic mutant. csp mutant embryos have an increased mitotic index, have highly disorganized mitotic spindles, and often lack one or both centrosomes. These phenotypes are caused by a loss-of-function mutation in zebrafish sil. To determine if the requirement for SIL in mitotic spindle organization is conserved in mammals, we generated an antibody against human SIL, which revealed that SIL localizes to the poles of the mitotic spindle during metaphase. Furthermore, short hairpin RNA knockdown of SIL in human cells recapitulates the zebrafish csp mitotic spindle defects. These data, taken together, identify SIL as a novel, vertebrate-specific regulator of mitotic spindle assembly. PMID:17576815

  13. Bayesian Classification and Regression with High Dimensional Features

    NASA Astrophysics Data System (ADS)

    Li, Longhai

    2007-09-01

    This thesis responds to the challenges of using a large number, such as thousands, of features in regression and classification problems. There are two situations where such high dimensional features arise. One is when high dimensional measurements are available, for example, gene expression data produced by microarray techniques. For computational or other reasons, people may select only a small subset of features when modelling such data, by looking at how relevant the features are to predicting the response, based on some measure such as correlation with the response in the training data. Although it is used very commonly, this procedure will make the response appear more predictable than it actually is. In Chapter 2, we propose a Bayesian method to avoid this selection bias, with application to naive Bayes models and mixture models. High dimensional features also arise when we consider high-order interactions. The number of parameters will increase exponentially with the order considered. In Chapter 3, we propose a method for compressing a group of parameters into a single one, by exploiting the fact that many predictor variables derived from high-order interactions have the same values for all the training cases. The number of compressed parameters may have converged before considering the highest possible order. We apply this compression method to logistic sequence prediction models and logistic classification models. We use both simulated data and real data to test our methods in both chapters.

  14. A mitotic function for the high-mobility group protein HMG20b regulated by its interaction with the BRC repeats of the BRCA2 tumor suppressor.

    PubMed

    Lee, M; Daniels, M J; Garnett, M J; Venkitaraman, A R

    2011-07-28

    The inactivation of BRCA2, a suppressor of breast, ovarian and other epithelial cancers, triggers instability in chromosome structure and number, which are thought to arise from defects in DNA recombination and mitotic cell division, respectively. Human BRCA2 controls DNA recombination via eight BRC repeats, evolutionarily conserved motifs of ∼35 residues, that interact directly with the recombinase RAD51. How BRCA2 controls mitotic cell division is debated. Several studies by different groups report that BRCA2 deficiency affects cytokinesis. Moreover, its interaction with HMG20b, a protein of uncertain function containing a promiscuous DNA-binding domain and kinesin-like coiled coils, has been implicated in the G2-M transition. We show here that HMG20b depletion by RNA interference disturbs the completion of cell division, suggesting a novel function for HMG20b. In vitro, HMG20b binds directly to the BRC repeats of BRCA2, and exhibits the highest affinity for BRC5, a motif that binds poorly to RAD51. Conversely, the BRC4 repeat binds strongly to RAD51, but not to HMG20b. In vivo, BRC5 overexpression inhibits the BRCA2-HMG20b interaction, recapitulating defects in the completion of cell division provoked by HMG20b depletion. In contrast, BRC4 inhibits the BRCA2-RAD51 interaction and the assembly of RAD51 at sites of DNA damage, but not the completion of cell division. Our findings suggest that a novel function for HMG20b in cytokinesis is regulated by its interaction with the BRC repeats of BRCA2, and separate this unexpected function for the BRC repeats from their known activity in DNA recombination. We propose that divergent tumor-suppressive pathways regulating chromosome segregation as well as chromosome structure may be governed by the conserved BRC motifs in BRCA2. PMID:21399666

  15. How do anti-mitotic drugs kill cancer cells?

    PubMed

    Gascoigne, Karen E; Taylor, Stephen S

    2009-08-01

    In 2007, over 12-million people were diagnosed with cancer. According to the American Cancer Society, at least one third of these individuals are not expected to survive the disease, making cancer the second most prevalent cause of death worldwide. Systemic chemotherapy forms the mainstay of cancer treatment, and agents that disrupt mitotic spindle assembly - so called ;anti-mitotics' - are commonly used to treat a wide variety of cancers. Traditional anti-mitotic agents include the microtubule toxins such as taxol, other taxanes and the vinca alkaloids, all of which have proven successful in the clinic. However, patient response remains highly unpredictable, and drug resistance is common. In addition, toxicity is a problem. To address these limitations, a new generation of anti-mitotic drugs is being developed. As the first wave of these new agents enters clinical trails, much hope rests on their outcome. Meanwhile, significant attention is being focused on trying to predict which tumour types are likely to respond. In this Commentary, we outline recent advances in our understanding of how cancer cells respond to anti-mitotic drugs, and discuss the relevance of these studies to their use in the clinic. PMID:19625502

  16. Loops determine the mechanical properties of mitotic chromosomes

    NASA Astrophysics Data System (ADS)

    Zhang, Yang; Heermann, Dieter W.

    2013-03-01

    In mitosis, chromosomes undergo a condensation into highly compacted, rod-like objects. Many models have been put forward for the higher-order organization of mitotic chromosomes including radial loop and hierarchical folding models. Additionally, mechanical properties of mitotic chromosomes under different conditions were measured. However, the internal organization of mitotic chromosomes still remains unclear. Here we present a polymer model for mitotic chromosomes and show how chromatin loops play a major role for their mechanical properties. The key assumption of the model is the ability of the chromatin fibre to dynamically form loops with the help of binding proteins. Our results show that looping leads to a tight compaction and significantly increases the bending rigidity of chromosomes. Moreover, our qualitative prediction of the force elongation behaviour is close to experimental findings. This indicates that the internal structure of mitotic chromosomes is based on self-organization of the chromatin fibre. We also demonstrate how number and size of loops have a strong influence on the mechanical properties. We suggest that changes in the mechanical characteristics of chromosomes can be explained by an altered internal loop structure. YZ gratefully appreciates funding by the German National Academic Foundation (Studienstiftung des deutschen Volkes) and support by the Heidelberg Graduate School for Mathematical and Computational Methods in the Sciences (HGS MathComp).

  17. The Sparse MLE for Ultra-High-Dimensional Feature Screening

    PubMed Central

    Xu, Chen; Chen, Jiahua

    2014-01-01

    Feature selection is fundamental for modeling the high dimensional data, where the number of features can be huge and much larger than the sample size. Since the feature space is so large, many traditional procedures become numerically infeasible. It is hence essential to first remove most apparently non-influential features before any elaborative analysis. Recently, several procedures have been developed for this purpose, which include the sure-independent-screening (SIS) as a widely-used technique. To gain the computational efficiency, the SIS screens features based on their individual predicting power. In this paper, we propose a new screening method via the sparsity-restricted maximum likelihood estimator (SMLE). The new method naturally takes the joint effects of features in the screening process, which gives itself an edge to potentially outperform the existing methods. This conjecture is further supported by the simulation studies under a number of modeling settings. We show that the proposed method is screening consistent in the context of ultra-high-dimensional generalized linear models. PMID:25382886

  18. A quantitative atlas of mitotic phosphorylation

    PubMed Central

    Dephoure, Noah; Zhou, Chunshui; Villén, Judit; Beausoleil, Sean A.; Bakalarski, Corey E.; Elledge, Stephen J.; Gygi, Steven P.

    2008-01-01

    The eukaryotic cell division cycle is characterized by a sequence of orderly and highly regulated events resulting in the duplication and separation of all cellular material into two newly formed daughter cells. Protein phosphorylation by cyclin-dependent kinases (CDKs) drives this cycle. To gain further insight into how phosphorylation regulates the cell cycle, we sought to identify proteins whose phosphorylation is cell cycle regulated. Using stable isotope labeling along with a two-step strategy for phosphopeptide enrichment and high mass accuracy mass spectrometry, we examined protein phosphorylation in a human cell line arrested in the G1 and mitotic phases of the cell cycle. We report the identification of >14,000 different phosphorylation events, more than half of which, to our knowledge, have not been described in the literature, along with relative quantitative data for the majority of these sites. We observed >1,000 proteins with increased phosphorylation in mitosis including many known cell cycle regulators. The majority of sites on regulated phosphopeptides lie in [S/T]P motifs, the minimum required sequence for CDKs, suggesting that many of the proteins may be CDK substrates. Analysis of non-proline site-containing phosphopeptides identified two unique motifs that suggest there are at least two undiscovered mitotic kinases. PMID:18669648

  19. Origin of the high vlos feature in the Galactic bar

    NASA Astrophysics Data System (ADS)

    Aumer, Michael; Schönrich, Ralph

    2015-12-01

    We analyse a controlled N-body+smoothed particle hydrodynamics simulation of a growing disc galaxy within a non-growing, live dark halo. The disc is continuously fed with gas and star particles on near-circular orbits and develops a bar comparable in size to the one of the Milky Way (MW). We extract line-of-sight velocity vlos distributions from the model and compare it to data recently obtained from the Apache Point Observatory Galactic Evolution Experiment (APOGEE) survey which show distinct high-velocity features around vlos ˜ 200 km s-1. With an APOGEE-like selection function, but without any scaling nor adjustment, we find vlos distributions very similar to those in APOGEE. The stars that make up the high vlos features at positive longitudes l are preferentially young bar stars (age τ ≲ 2-3 Gyr) which move away from us along the rear side of the bar. At negative l, we find the corresponding low vlos feature from stars moving towards us. At l > 10 deg, the highest vlos stars are a mixture of bar and background disc stars which complicates the interpretation of observations. The main peak in vlos is dominated by fore- and background stars. At a given time, ˜40-50 per cent of high vlos stars occupy x1-like orbits, but a significant fraction are on more complex orbits. The observed feature is likely due to a population of dynamically cool, young stars formed from gas just outside the bar and subsequently captured by the growing bar. The high vlos features disappear at high latitudes |b| ≳ 2 deg which explains the non-detection of such features in other surveys.

  20. Spectral feature design in high dimensional multispectral data

    NASA Technical Reports Server (NTRS)

    Chen, Chih-Chien Thomas; Landgrebe, David A.

    1988-01-01

    The High resolution Imaging Spectrometer (HIRIS) is designed to acquire images simultaneously in 192 spectral bands in the 0.4 to 2.5 micrometers wavelength region. It will make possible the collection of essentially continuous reflectance spectra at a spectral resolution sufficient to extract significantly enhanced amounts of information from return signals as compared to existing systems. The advantages of such high dimensional data come at a cost of increased system and data complexity. For example, since the finer the spectral resolution, the higher the data rate, it becomes impractical to design the sensor to be operated continuously. It is essential to find new ways to preprocess the data which reduce the data rate while at the same time maintaining the information content of the high dimensional signal produced. Four spectral feature design techniques are developed from the Weighted Karhunen-Loeve Transforms: (1) non-overlapping band feature selection algorithm; (2) overlapping band feature selection algorithm; (3) Walsh function approach; and (4) infinite clipped optimal function approach. The infinite clipped optimal function approach is chosen since the features are easiest to find and their classification performance is the best. After the preprocessed data has been received at the ground station, canonical analysis is further used to find the best set of features under the criterion that maximal class separability is achieved. Both 100 dimensional vegetation data and 200 dimensional soil data were used to test the spectral feature design system. It was shown that the infinite clipped versions of the first 16 optimal features had excellent classification performance. The overall probability of correct classification is over 90 percent while providing for a reduced downlink data rate by a factor of 10.

  1. Unique Features of a Highly Pathogenic Campylobacter jejuni Strain†

    PubMed Central

    Hofreuter, Dirk; Tsai, Jennifer; Watson, Robert O.; Novik, Veronica; Altman, Bill; Benitez, Michelle; Clark, Christina; Perbost, Clotilde; Jarvie, Thomas; Du, Lei; Galán, Jorge E.

    2006-01-01

    Campylobacter jejuni, a major human enteric pathogen, exhibits significant strain-to-strain differences which result in differences in pathogenic potential. C. jejuni 81-176 is a highly virulent strain that exhibits unique pathogenic features and is used by many research laboratories. We have determined the nucleotide sequence of its genome and compared it to the genomes of other sequenced C. jejuni strains. We identified a number of unique genetic features which may confer specific metabolic and pathogenic properties on this strain. We have also identified regions of the C. jejuni genome that are hot spots for the integration of horizontally acquired genetic material. This information should help the understanding of the pathogenesis of C. jejuni and, in particular, the unique features of this highly pathogenic strain. PMID:16861657

  2. Mitotic Spindle Disruption by Alternating Electric Fields Leads to Improper Chromosome Segregation and Mitotic Catastrophe in Cancer Cells.

    PubMed

    Giladi, Moshe; Schneiderman, Rosa S; Voloshin, Tali; Porat, Yaara; Munster, Mijal; Blat, Roni; Sherbo, Shay; Bomzon, Zeev; Urman, Noa; Itzhaki, Aviran; Cahal, Shay; Shteingauz, Anna; Chaudhry, Aafia; Kirson, Eilon D; Weinberg, Uri; Palti, Yoram

    2015-01-01

    Tumor Treating Fields (TTFields) are low intensity, intermediate frequency, alternating electric fields. TTFields are a unique anti-mitotic treatment modality delivered in a continuous, noninvasive manner to the region of a tumor. It was previously postulated that by exerting directional forces on highly polar intracellular elements during mitosis, TTFields could disrupt the normal assembly of spindle microtubules. However there is limited evidence directly linking TTFields to an effect on microtubules. Here we report that TTFields decrease the ratio between polymerized and total tubulin, and prevent proper mitotic spindle assembly. The aberrant mitotic events induced by TTFields lead to abnormal chromosome segregation, cellular multinucleation, and caspase dependent apoptosis of daughter cells. The effect of TTFields on cell viability and clonogenic survival substantially depends upon the cell division rate. We show that by extending the duration of exposure to TTFields, slowly dividing cells can be affected to a similar extent as rapidly dividing cells. PMID:26658786

  3. Mitotic Spindle Disruption by Alternating Electric Fields Leads to Improper Chromosome Segregation and Mitotic Catastrophe in Cancer Cells

    PubMed Central

    Giladi, Moshe; Schneiderman, Rosa S; Voloshin, Tali; Porat, Yaara; Munster, Mijal; Blat, Roni; Sherbo, Shay; Bomzon, Zeev; Urman, Noa; Itzhaki, Aviran; Cahal, Shay; Shteingauz, Anna; Chaudhry, Aafia; Kirson, Eilon D; Weinberg, Uri; Palti, Yoram

    2015-01-01

    Tumor Treating Fields (TTFields) are low intensity, intermediate frequency, alternating electric fields. TTFields are a unique anti-mitotic treatment modality delivered in a continuous, noninvasive manner to the region of a tumor. It was previously postulated that by exerting directional forces on highly polar intracellular elements during mitosis, TTFields could disrupt the normal assembly of spindle microtubules. However there is limited evidence directly linking TTFields to an effect on microtubules. Here we report that TTFields decrease the ratio between polymerized and total tubulin, and prevent proper mitotic spindle assembly. The aberrant mitotic events induced by TTFields lead to abnormal chromosome segregation, cellular multinucleation, and caspase dependent apoptosis of daughter cells. The effect of TTFields on cell viability and clonogenic survival substantially depends upon the cell division rate. We show that by extending the duration of exposure to TTFields, slowly dividing cells can be affected to a similar extent as rapidly dividing cells. PMID:26658786

  4. Axin localizes to mitotic spindles and centrosomes in mitotic cells

    SciTech Connect

    Kim, Shi-Mun; Choi, Eun-Jin; Song, Ki-Joon; Kim, Sewoon; Seo, Eunjeong; Jho, Eek-Hoon; Kee, Sun-Ho

    2009-04-01

    Wnt signaling plays critical roles in cell proliferation and carcinogenesis. In addition, numerous recent studies have shown that various Wnt signaling components are involved in mitosis and chromosomal instability. However, the role of Axin, a negative regulator of Wnt signaling, in mitosis has remained unclear. Using monoclonal antibodies against Axin, we found that Axin localizes to the centrosome and along mitotic spindles. This localization was suppressed by siRNA specific for Aurora A kinase and by Aurora kinase inhibitor. Interestingly, Axin over-expression altered the subcellular distribution of Plk1 and of phosphorylated glycogen synthase kinase (GSK3{beta}) without producing any notable changes in cellular phenotype. In the presence of Aurora kinase inhibitor, Axin over-expression induced the formation of cleavage furrow-like structures and of prominent astral microtubules lacking midbody formation in a subset of cells. Our results suggest that Axin modulates distribution of Axin-associated proteins such as Plk1 and GSK3{beta} in an expression level-dependent manner and these interactions affect the mitotic process, including cytokinesis under certain conditions, such as in the presence of Aurora kinase inhibitor.

  5. Mitotic force generators and chromosome segregation

    PubMed Central

    Civelekoglu-Scholey, Gul

    2010-01-01

    The mitotic spindle uses dynamic microtubules and mitotic motors to generate the pico-Newton scale forces that are needed to drive the mitotic movements that underlie chromosome capture, alignment and segregation. Here, we consider the biophysical and molecular basis of force-generation for chromosome movements in the spindle, and, with reference to the Drosophila embryo mitotic spindle, we briefly discuss how mathematical modeling can complement experimental analysis to illuminate the mechanisms of chromosome-to-pole motility during anaphase A and spindle elongation during anaphase B. PMID:20221784

  6. Feature extraction and classification algorithms for high dimensional data

    NASA Technical Reports Server (NTRS)

    Lee, Chulhee; Landgrebe, David

    1993-01-01

    Feature extraction and classification algorithms for high dimensional data are investigated. Developments with regard to sensors for Earth observation are moving in the direction of providing much higher dimensional multispectral imagery than is now possible. In analyzing such high dimensional data, processing time becomes an important factor. With large increases in dimensionality and the number of classes, processing time will increase significantly. To address this problem, a multistage classification scheme is proposed which reduces the processing time substantially by eliminating unlikely classes from further consideration at each stage. Several truncation criteria are developed and the relationship between thresholds and the error caused by the truncation is investigated. Next an approach to feature extraction for classification is proposed based directly on the decision boundaries. It is shown that all the features needed for classification can be extracted from decision boundaries. A characteristic of the proposed method arises by noting that only a portion of the decision boundary is effective in discriminating between classes, and the concept of the effective decision boundary is introduced. The proposed feature extraction algorithm has several desirable properties: it predicts the minimum number of features necessary to achieve the same classification accuracy as in the original space for a given pattern recognition problem; and it finds the necessary feature vectors. The proposed algorithm does not deteriorate under the circumstances of equal means or equal covariances as some previous algorithms do. In addition, the decision boundary feature extraction algorithm can be used both for parametric and non-parametric classifiers. Finally, some problems encountered in analyzing high dimensional data are studied and possible solutions are proposed. First, the increased importance of the second order statistics in analyzing high dimensional data is recognized. By investigating the characteristics of high dimensional data, the reason why the second order statistics must be taken into account in high dimensional data is suggested. Recognizing the importance of the second order statistics, there is a need to represent the second order statistics. A method to visualize statistics using a color code is proposed. By representing statistics using color coding, one can easily extract and compare the first and the second statistics.

  7. Identifying high-level features of texture perception

    NASA Astrophysics Data System (ADS)

    Rao, A. Ravishankar; Lohse, Gerald L.

    1992-08-01

    A fundamental issue in texture analysis is that of deciding what textural features are important in texture perception, and how they are used. Experiments on human pre-attentive vision have identified several low-level features (such as orientation on blobs, and size of line segments), which are used in texture perception. However, the question of what higher level features of texture are used has not been adequately addressed. We designed an experiment to help identify the relevant higher order features of texture perceived by humans. We used twenty subjects, who were asked to perform an unsupervised classification of thirty pictures from Brodatz's album on texture. Each subject was asked to group these pictures into as many classes as desired. Both hierarchical cluster analysis and non-metric MDS were applied to the pooled similarity matrix generated from the subjects' groupings. A surprising outcome is that the MDS solutions fit the data very well. The stress in the two dimensional case is 0.10, and in the three dimensional case is 0.045. We rendered the original textures in these coordinate systems, and interpreted the (rotated) axes. It appears that the axes in the 2D case correspond to periodicity versus irregularity, and directional versus non-directional. In the 3D case, the third dimension represents the structural complexity of the texture. Furthermore, the clusters identified by the hierarchical cluster analysis remain virtually intact in the MDS solution. The results of our experiment indicate that people use three high level features for texture perception. Future studies are needed to determine the appropriateness of these high-level features for computational texture analysis and classification.

  8. Prognostic comparison of the proliferation markers (mitotic activity index, phosphohistone H3, Ki67), steroid receptors, HER2, high molecular weight cytokeratins and classical prognostic factors in T₁₋₂N₀M₀ breast cancer.

    PubMed

    Gudlaugsson, Einar; Klos, Jan; Skaland, Ivar; Janssen, Emiel A M; Smaaland, Rune; Feng, Weiwei; Shao, Zhimin; Malpica, Anais; Baak, Jan P A

    2013-04-01

    The proliferation factors: mitotic activity index (MAI), phosphohistone H3 (PPH3) and Ki67 have strong prognostic value in early breast cancer but their independent value to each other and other prognostic factors has not been evaluated. In 237 T₁₋₂N₀M₀ breast cancers without systemic adjuvant treatment, formalized MAI assessment and strictly standardized, fully automated quantitative immunohistochemistry (IHC) for Ki67, PPH3, estrogen (ER) and progesterone receptor (PR), HER2, cytokeratins-5/6 and -14, and automated digital image analysis (DIA) for measuring PPH3 and Ki67 were performed. Section thickness was measured to further control IHC measurements. All features were measured in the periphery of tumors. The different proliferation assessments and other well-established clinicopathological and biomarker prognostic factors were compared. DIA-Ki67 added prognostically to PPH3. None of the other biomarkers or clinicopathological variables added prognostically to this PPH3/Ki67 combination. However, when PPH3 is replaced by MAI the prognostic value is nearly the same. In early operable node negative breast cancer without adjuvant systemic treatment, Ki67 with a threshold of 6.5% assessed by digital image analysis in the periphery of the tumor is prognostically strong. The combination of either PPH3/Ki67 or MAI/Ki67 overshadowed the prognostic value of all other features including Ki67 alone. PMID:23625593

  9. The nucleoporin ALADIN regulates Aurora A localization to ensure robust mitotic spindle formation

    PubMed Central

    Carvalhal, Sara; Ribeiro, Susana Abreu; Arocena, Miguel; Kasciukovic, Taciana; Temme, Achim; Koehler, Katrin; Huebner, Angela; Griffis, Eric R.

    2015-01-01

    The formation of the mitotic spindle is a complex process that requires massive cellular reorganization. Regulation by mitotic kinases controls this entire process. One of these mitotic controllers is Aurora A kinase, which is itself highly regulated. In this study, we show that the nuclear pore protein ALADIN is a novel spatial regulator of Aurora A. Without ALADIN, Aurora A spreads from centrosomes onto spindle microtubules, which affects the distribution of a subset of microtubule regulators and slows spindle assembly and chromosome alignment. ALADIN interacts with inactive Aurora A and is recruited to the spindle pole after Aurora A inhibition. Of interest, mutations in ALADIN cause triple A syndrome. We find that some of the mitotic phenotypes that we observe after ALADIN depletion also occur in cells from triple A syndrome patients, which raises the possibility that mitotic errors may underlie part of the etiology of this syndrome. PMID:26246606

  10. OTSSP167 Abrogates Mitotic Checkpoint through Inhibiting Multiple Mitotic Kinases.

    PubMed

    Ji, Wenbin; Arnst, Christopher; Tipton, Aaron R; Bekier, Michael E; Taylor, William R; Yen, Tim J; Liu, Song-Tao

    2016-01-01

    OTSSP167 was recently characterized as a potent inhibitor for maternal embryonic leucine zipper kinase (MELK) and is currently tested in Phase I clinical trials for solid tumors that have not responded to other treatment. Here we report that OTSSP167 abrogates the mitotic checkpoint at concentrations used to inhibit MELK. The abrogation is not recapitulated by RNAi mediated silencing of MELK in cells. Although OTSSP167 indeed inhibits MELK, it exhibits off-target activity against Aurora B kinase in vitro and in cells. Furthermore, OTSSP167 inhibits BUB1 and Haspin kinases, reducing phosphorylation at histones H2AT120 and H3T3 and causing mislocalization of Aurora B and associated chromosomal passenger complex from the centromere/kinetochore. The results suggest that OTSSP167 may have additional mechanisms of action for cancer cell killing and caution the use of OTSSP167 as a MELK specific kinase inhibitor in biochemical and cellular assays. PMID:27082996

  11. OTSSP167 Abrogates Mitotic Checkpoint through Inhibiting Multiple Mitotic Kinases

    PubMed Central

    Tipton, Aaron R.; Bekier, Michael E.; Taylor, William R.; Yen, Tim J.; Liu, Song-Tao

    2016-01-01

    OTSSP167 was recently characterized as a potent inhibitor for maternal embryonic leucine zipper kinase (MELK) and is currently tested in Phase I clinical trials for solid tumors that have not responded to other treatment. Here we report that OTSSP167 abrogates the mitotic checkpoint at concentrations used to inhibit MELK. The abrogation is not recapitulated by RNAi mediated silencing of MELK in cells. Although OTSSP167 indeed inhibits MELK, it exhibits off-target activity against Aurora B kinase in vitro and in cells. Furthermore, OTSSP167 inhibits BUB1 and Haspin kinases, reducing phosphorylation at histones H2AT120 and H3T3 and causing mislocalization of Aurora B and associated chromosomal passenger complex from the centromere/kinetochore. The results suggest that OTSSP167 may have additional mechanisms of action for cancer cell killing and caution the use of OTSSP167 as a MELK specific kinase inhibitor in biochemical and cellular assays. PMID:27082996

  12. AMPK and PFKFB3 mediate glycolysis and survival in response to mitophagy during mitotic arrest.

    PubMed

    Doménech, Elena; Maestre, Carolina; Esteban-Martínez, Lorena; Partida, David; Pascual, Rosa; Fernández-Miranda, Gonzalo; Seco, Esther; Campos-Olivas, Ramón; Pérez, Manuel; Megias, Diego; Allen, Katherine; López, Miguel; Saha, Asish K; Velasco, Guillermo; Rial, Eduardo; Méndez, Raúl; Boya, Patricia; Salazar-Roa, María; Malumbres, Marcos

    2015-10-01

    Blocking mitotic progression has been proposed as an attractive therapeutic strategy to impair proliferation of tumour cells. However, how cells survive during prolonged mitotic arrest is not well understood. We show here that survival during mitotic arrest is affected by the special energetic requirements of mitotic cells. Prolonged mitotic arrest results in mitophagy-dependent loss of mitochondria, accompanied by reduced ATP levels and the activation of AMPK. Oxidative respiration is replaced by glycolysis owing to AMPK-dependent phosphorylation of PFKFB3 and increased production of this protein as a consequence of mitotic-specific translational activation of its mRNA. Induction of autophagy or inhibition of AMPK or PFKFB3 results in enhanced cell death in mitosis and improves the anti-tumoral efficiency of microtubule poisons in breast cancer cells. Thus, survival of mitotic-arrested cells is limited by their metabolic requirements, a feature with potential implications in cancer therapies aimed to impair mitosis or metabolism in tumour cells. PMID:26322680

  13. Mitotic and mitogenic Wnt signalling

    PubMed Central

    Niehrs, Christof; Acebron, Sergio P

    2012-01-01

    Canonical Wnt signalling plays an important role in development, tissue homeostasis, and cancer. At the cellular level, canonical Wnt signalling acts by regulating cell fate, cell growth, and cell proliferation. With regard to proliferation, there is increasing evidence for a complex interaction between canonical Wnt signalling and the cell cycle. Mitogenic Wnt signalling regulates cell proliferation by promoting G1 phase. In mitosis, components of the Wnt signalling cascade function directly in spindle formation. Moreover, Wnt signalling is strongly activated in mitosis, suggesting that ‘mitotic Wnt signalling' plays an important role to orchestrate a cell division program. Here, we review the complex interplay between Wnt signalling and the cell cycle. PMID:22617425

  14. High Spatial Velocity Features in the Orion Nebula,

    NASA Astrophysics Data System (ADS)

    O'Dell, C. R.; Henney, W. J.

    2008-10-01

    We have used widely spaced in time Hubble Space Telescope images to determine tangential velocities of features associated with outflows from young stars. These observations were supplemented by ground-based telescope spectroscopy, and from the resultant radial velocities, space velocities were determined for many outflows. Numerous new moving features were found and grouped into known and newly assigned Herbig-Haro objects. It was found that stellar outflow is highly discontinuous, as frequently is the case, with long-term gaps of a few hundred years, and that these outflow periods are marked by staccato bursts over periods of about ten years. Although this has been observed in other regions, the Orion Nebula Cluster presents the richest display of this property. Most of the large-scale Herbig-Haro objects in the brightest part of the Orion Nebula appear to originate from a small region northeast of the strong Orion-S radio and infrared sources. With the possible exception of HH 203, we are not able to identify specific stellar sources, but do identify candidate sources for several other bright Herbig-Haro objects. We find that there are optical features in the BN-KL region that can be related to the known large-scale outflow that originates there. We find additional evidence for this outflow originating 500-1000 years ago. Based on observations at the San Pedro Martir Observatory operated by the Universidad Nacional Autónoma de México.

  15. High resolution cloud feature tracking on Venus by Galileo

    NASA Technical Reports Server (NTRS)

    Toigo, Anthony; Gierasch, Peter J.; Smith, Michael D.

    1994-01-01

    The Venus cloud deck was monitored in February 1990 for 16 hours at 400 nanometers wavelength by the Galileo imaging system, with a spatial resolution of about 15 km and with image time separations as small as 10 minutes. Velocities are deduced by following the motion of small cloud features. In spite of the high temporal frequence is capable of being detected, no dynamical phenomena are apparent in the velocity data except the already well-known solar tides, possibly altered by the slow 4-day wave and the Hadley circulation. There is no evidence, to a level of approximately 4 m/s, of eddy or wavelike activity. The dominant size of sub-global scale albedo features is 200-500 km, and their contrast is approximately 5%. At low altitudes there are patches of blotchy, cell-like structures but at most locations the markings are streaky. The patterns are similar to those discovered by Mariner 10 and Pioneer Venus (M. J. S. Belton et al., 1976, W. B. Rossow et al., 1980). Scaling arguments are presented to argue that the mesoscale blotchy cell-like cloud patterns are caused by local dynamics driven in a shallow layer by differential absorption of sunlight. It is also argued that mesoscale albedo features are either streaky or cell-like simply depending on whether the horizontal shear of the large scale flow exceeds a certain critical value.

  16. Efficient learning and feature selection in high-dimensional regression.

    PubMed

    Ting, Jo-Anne; D'Souza, Aaron; Vijayakumar, Sethu; Schaal, Stefan

    2010-04-01

    We present a novel algorithm for efficient learning and feature selection in high-dimensional regression problems. We arrive at this model through a modification of the standard regression model, enabling us to derive a probabilistic version of the well-known statistical regression technique of backfitting. Using the expectation-maximization algorithm, along with variational approximation methods to overcome intractability, we extend our algorithm to include automatic relevance detection of the input features. This variational Bayesian least squares (VBLS) approach retains its simplicity as a linear model, but offers a novel statistically robust black-box approach to generalized linear regression with high-dimensional inputs. It can be easily extended to nonlinear regression and classification problems. In particular, we derive the framework of sparse Bayesian learning, the relevance vector machine, with VBLS at its core, offering significant computational and robustness advantages for this class of methods. The iterative nature of VBLS makes it most suitable for real-time incremental learning, which is crucial especially in the application domain of robotics, brain-machine interfaces, and neural prosthetics, where real-time learning of models for control is needed. We evaluate our algorithm on synthetic and neurophysiological data sets, as well as on standard regression and classification benchmark data sets, comparing it with other competitive statistical approaches and demonstrating its suitability as a drop-in replacement for other generalized linear regression techniques. PMID:20028222

  17. [Metabolism features of bacteria resistant to high concentrations of chromate].

    PubMed

    Smirnova, G F; Podgorskiĭ, V S

    2013-01-01

    Twenty strains of bacteria resistant to high concentrations of chromate were isolated from different ecological niches. They were able to reduce chromate to compounds of trivalent chromium--nonsoluble chromium hydroxide or soluble crystalline hydrates of trivalent chromium. The growth features of these microorganisms on media containing chromate at high concentrations (up to 20.0 g/l) are described. Besides chromate bacteria can reduce vanadate to compounds of V(4+) and Mo(6+) to Mo(5+). The best reduction takes place on the media where MPB. glucose or ethanol serves as the source of carbon. The growth and reduction of anion-in-study did not occur on organic acids. It was shown that tungstate, chlorate or perchlorate were not toxic for the studied bacteria up to concentrations of 10.0 g/l, however were not reduced by these microorganisms. The most active strains belong to genera Pseudomonas, Oerskovia, Bacillus, Micrococcus. PMID:23720958

  18. Efficient Activation of Apoptotic Signaling during Mitotic Arrest with AK301

    PubMed Central

    Bleiler, Marina; Yeagley, Michelle; Wright, Dennis; Giardina, Charles

    2016-01-01

    Mitotic inhibitors are widely utilized chemotherapeutic agents that take advantage of mitotic defects in cancer cells. We have identified a novel class of piperazine-based mitotic inhibitors, of which AK301 is the most potent derivative identified to date (EC50 < 200 nM). Colon cancer cells arrested in mitosis with AK301 readily underwent a p53-dependent apoptosis following compound withdrawal and arrest release. This apoptotic response was significantly higher for AK301 than for other mitotic inhibitors tested (colchicine, vincristine, and BI 2536). AK301-treated cells exhibited a robust mitosis-associated DNA damage response, including ATM activation, γH2AX phosphorylation and p53 stabilization. The association between mitotic signaling and the DNA damage response was supported by the finding that Aurora B inhibition reduced the level of γH2AX staining. Confocal imaging of AK301-treated cells revealed multiple γ-tubulin microtubule organizing centers attached to microtubules, but with limited centrosome migration, raising the possibility that aberrant microtubule pulling may underlie DNA breakage. AK301 selectively targeted APC-mutant colonocytes and promoted TNF-induced apoptosis in p53-mutant colon cancer cells. Our findings indicate that AK301 induces a mitotic arrest state with a highly active DNA damage response. Together with a reversible arrest state, AK301 is a potent promoter of a mitosis-to-apoptosis transition that can target cancer cells with mitotic defects. PMID:27097159

  19. The chromosomal passenger complex (CPC) as a key orchestrator of orderly mitotic exit and cytokinesis

    PubMed Central

    Kitagawa, Mayumi; Lee, Sang Hyun

    2015-01-01

    Understanding the molecular network of orderly mitotic exit to re-establish a functional interphase nucleus is critical because disordered mitotic exit inevitably leads to genomic instability. In contrast to the mechanisms of the entrance to mitosis, however, little is known about what controls the orderly exit from mitosis, particularly in mammalian cells. The chromosomal passenger complex (CPC), which is composed of Aurora B, INCENP, Borealin and Survivin, is one of the most widely studied and highly conserved hetero-tetrameric complexes. The CPC orchestrates proper chromosome segregation with cytokinesis by targeting to specific locations at different stages of mitosis. Recent studies reveal that controlling CPC localization and Aurora B kinase activity also serves as a key surveillance mechanism for the orderly mitotic exit. This ensures the reformation of a functional interphase nucleus from condensed mitotic chromosomes by delaying mitotic exit and cytokinetic processes in response to defects in chromosome segregation. In this review, we will summarize the latest insight into the molecular mechanisms that regulate CPC localization during mitotic exit and discuss how targeting Aurora B activity to different locations at different times impacts executing multiple mitotic exit events in order and recently proposed surveillance mechanisms. Finally, we briefly discuss the potential implication of deregulated Aurora B in inducing genomic damage and tumorigenesis with current efforts in targeting Aurora B activity for anti-cancer therapy. PMID:25798441

  20. On the molecular mechanisms of mitotic kinase activation.

    PubMed

    Bayliss, Richard; Fry, Andrew; Haq, Tamanna; Yeoh, Sharon

    2012-11-01

    During mitosis, human cells exhibit a peak of protein phosphorylation that alters the behaviour of a significant proportion of proteins, driving a dramatic transformation in the cell's shape, intracellular structures and biochemistry. These mitotic phosphorylation events are catalysed by several families of protein kinases, including Auroras, Cdks, Plks, Neks, Bubs, Haspin and Mps1/TTK. The catalytic activities of these kinases are activated by phosphorylation and through protein-protein interactions. In this review, we summarize the current state of knowledge of the structural basis of mitotic kinase activation mechanisms. This review aims to provide a clear and comprehensive primer on these mechanisms to a broad community of researchers, bringing together the common themes, and highlighting specific differences. Along the way, we have uncovered some features of these proteins that have previously gone unreported, and identified unexplored questions for future work. The dysregulation of mitotic kinases is associated with proliferative disorders such as cancer, and structural biology will continue to play a critical role in the development of chemical probes used to interrogate disease biology and applied to the treatment of patients. PMID:23226601

  1. On the molecular mechanisms of mitotic kinase activation

    PubMed Central

    Bayliss, Richard; Fry, Andrew; Haq, Tamanna; Yeoh, Sharon

    2012-01-01

    During mitosis, human cells exhibit a peak of protein phosphorylation that alters the behaviour of a significant proportion of proteins, driving a dramatic transformation in the cell's shape, intracellular structures and biochemistry. These mitotic phosphorylation events are catalysed by several families of protein kinases, including Auroras, Cdks, Plks, Neks, Bubs, Haspin and Mps1/TTK. The catalytic activities of these kinases are activated by phosphorylation and through protein–protein interactions. In this review, we summarize the current state of knowledge of the structural basis of mitotic kinase activation mechanisms. This review aims to provide a clear and comprehensive primer on these mechanisms to a broad community of researchers, bringing together the common themes, and highlighting specific differences. Along the way, we have uncovered some features of these proteins that have previously gone unreported, and identified unexplored questions for future work. The dysregulation of mitotic kinases is associated with proliferative disorders such as cancer, and structural biology will continue to play a critical role in the development of chemical probes used to interrogate disease biology and applied to the treatment of patients. PMID:23226601

  2. Orthologues of the Anaphase-Promoting Complex/Cyclosome Coactivators Cdc20p and Cdh1p Are Important for Mitotic Progression and Morphogenesis in Candida albicans ?

    PubMed Central

    Chou, Hsini; Glory, Amandeep; Bachewich, Catherine

    2011-01-01

    The conserved anaphase-promoting complex/cyclosome (APC/C) system mediates protein degradation during mitotic progression. Conserved coactivators Cdc20p and Cdh1p regulate the APC/C during early to late mitosis and G1 phase. Candida albicans is an important fungal pathogen of humans, and it forms highly polarized cells when mitosis is blocked through depletion of the polo-like kinase Cdc5p or other treatments. However, the mechanisms governing mitotic progression and associated polarized growth in the pathogen are poorly understood. In order to gain insights into these processes, we characterized C. albicans orthologues of Cdc20p and Cdh1p. Cdc20p-depleted cells were blocked in early or late mitosis with elevated levels of Cdc5p and the mitotic cyclin Clb2p, suggesting that Cdc20p is essential and has some conserved functions during mitosis. However, the yeast cells formed highly polarized buds in contrast to the large doublets of S. cerevisiae cdc20 mutants, implying a distinct role in morphogenesis. In comparison, cdh1?/cdh1? cells were viable but showed enrichment of Clb2p and Cdc5p, suggesting that Cdh1p may influence mitotic exit. The cdh1?/cdh1? phenotype was pleiotropic, consisting of normal or enlarged yeast, pseudohyphae, and some elongated buds, whereas S. cerevisiae cdh1? yeast cells were reduced in size. Thus, C. albicans Cdh1p may have some distinct functions. Finally, absence of Cdh1p or Cdc20p had a minor or no effect on hyphal development, respectively. Overall, the results suggest that Cdc20p and Cdh1p may be APC/C activators that are important for mitosis but also morphogenesis in C. albicans. Their novel features imply additional variations in function and underscore rewiring in the emerging mitotic regulatory networks of the pathogen. PMID:21398510

  3. Practical multi-featured perfect absorber utilizing high conductivity silicon

    NASA Astrophysics Data System (ADS)

    Gok, Abdullah; Yilmaz, Mehmet; Bıyıklı, Necmi; Topallı, Kağan; Okyay, Ali K.

    2016-03-01

    We designed all-silicon, multi-featured band-selective perfect absorbing surfaces based on CMOS compatible processes. The center wavelength of the band-selective absorber can be varied between 2 and 22 μm while a bandwidth as high as 2.5 μm is demonstrated. We used a silicon-on-insulator (SOI) wafer which consists of n-type silicon (Si) device layer, silicon dioxide (SiO2) as buried oxide layer, and n-type Si handle layer. The center wavelength and bandwidth can be tuned by adjusting the conductivity of the Si device and handle layers as well as the thicknesses of the device and buried oxide layers. We demonstrate proof-of-concept absorber surfaces experimentally. Such absorber surfaces are easy to microfabricate because the absorbers do not require elaborate microfabrication steps such as patterning. Due to the structural simplicity, low-cost fabrication, wide spectrum range of operation, and band properties of the perfect absorber, the proposed multi-featured perfect absorber surfaces are promising for many applications. These include sensing devices, surface enhanced infrared absorption applications, solar cells, meta-materials, frequency selective sensors and modulators.

  4. Features of photoconversion in highly efficient silicon solar cells

    SciTech Connect

    Sachenko, A. V.; Shkrebtii, A. I.; Korkishko, R. M.; Kostylyov, V. P.; Kulish, N. R.; Sokolovskyi, I. O.

    2015-02-15

    The photoconversion efficiency η in highly efficient silicon-based solar cells (SCs) is analyzed depending on the total surface-recombination rate S{sub s} on illuminated and rear surfaces. Solar cells based on silicon p-n junctions and α-Si:H or α-SiC:H-Si heterojunctions (so-called HIT structures) are considered in a unified approach. It is shown that a common feature of these SCs is an increased open-circuit voltage V{sub oc} associated with an additional contribution of the rear surface. Within an approach based on analysis of the physical features of photoconversion in SCs, taking into account the main recombination mechanisms, including Shockley-Read-Hall recombination, radiative recombination, surface recombination, recombination in the space-charge region, and band-to-band Auger recombination, expressions for the photoconversion efficiency of such SCs are obtained. The developed theory is compared with experiments, including those for SCs with record parameters, e.g., η = 25% and 24.7% for SCs with a p-n junction for HIT structures, respectively, under AM1.5 conditions. By comparing theory and experiment, the values of S{sub s} achieved as a result of recombination-loss minimization by various methods are determined. The results of calculations of the maximum possible value η{sub max} in silicon SCs are compared with the data of other papers. Good agreement is observed.

  5. Radar-anomalous, high-altitude features on Venus

    NASA Technical Reports Server (NTRS)

    Muhleman, Duane O.; Butler, Bryan J.

    1992-01-01

    Over nearly all of the surface of Venus the reflectivity and emissivity at centimeter wavelengths are about 0.15 and 0.85 respectively. These values are consistent with moderately dense soils and rock populations, but the mean reflectivity is about a factor of 2 greater than that for the Moon and other terrestrial planets. Pettingill and Ford, using Pioneer Venus reflectivities and emissivities, found a number of anomalous features on Venus that showed much higher reflectivities and much lower emissivities with both values approaching 0.5. These include Maxwell Montes, a number of high regions in Aphrodite Terra and Beta Regio, and several isolated mountain peaks. Most of the features are at altitudes above the mean radius by 2 to 3 km or more. However, such features have been found in the Magellan data at low altitudes and the anomalies do not exist on all high structures, Maat Mons being the most outstanding example. A number of papers have been written that attempt to explain the phenomena in terms of the geochemistry balance of weathering effects on likely surface minerals. The geochemists have shown that the fundamentally basaltic surface would be stable at the temperatures and pressures of the mean radius in the form of magnetite, but would evolve to pyrite and/or pyrrhotite in the presence of sulfur-bearing compounds such as SO2. Pyrite will be stable at altitudes above 4 or 5 km on Venus. Although the geochemical arguments are rather compelling, it is vitally important to rationally look at other explanations for radar and radio emission measurements such as that presented by Tryka and Muhleman. The radar reflectivity values are retrieved from the raw Magellan backscatter measurements by fitting the Hagfors' radar scattering model in which a surface roughness parameters and a normal incidence electrical reflectivity are estimated. The assumptions of the theory behind the model must be considered carefully before the results can be believed. These include that the surface roughness exists only at horizontal scales large compared to the wavelength, the vertical deviations are gaussianly distributed, there is no shadowing, and that the reflection occurs at the interface of two homogeneous dielectric half-spaces. Probably all these conditions are violated at the anomalous features under discussion. The most important of these is the homogeneity of the near surface of Venus, particularly in highlands. Under the assumptions of the theory, all of the radio energy is reflected by the impedance jump at the very boundary. However, in heterogeneous soil some fraction of the illuminating energy is propagated into the soil and then scattered back out by impedance discontinuities such as rock, voids, and cracks. In light soils, the latter effect can overwhelm the scattering effects of the true surface and greatly enhance the backscatter power, suggesting a much higher value of an effective dielectric constant that would be estimated from Hagfors' model.

  6. A comprehensive model to predict mitotic division in budding yeasts

    PubMed Central

    Sutradhar, Sabyasachi; Yadav, Vikas; Sridhar, Shreyas; Sreekumar, Lakshmi; Bhattacharyya, Dibyendu; Ghosh, Santanu Kumar; Paul, Raja; Sanyal, Kaustuv

    2015-01-01

    High-fidelity chromosome segregation during cell division depends on a series of concerted interdependent interactions. Using a systems biology approach, we built a robust minimal computational model to comprehend mitotic events in dividing budding yeasts of two major phyla: Ascomycota and Basidiomycota. This model accurately reproduces experimental observations related to spindle alignment, nuclear migration, and microtubule (MT) dynamics during cell division in these yeasts. The model converges to the conclusion that biased nucleation of cytoplasmic microtubules (cMTs) is essential for directional nuclear migration. Two distinct pathways, based on the population of cMTs and cortical dyneins, differentiate nuclear migration and spindle orientation in these two phyla. In addition, the model accurately predicts the contribution of specific classes of MTs in chromosome segregation. Thus we present a model that offers a wider applicability to simulate the effects of perturbation of an event on the concerted process of the mitotic cell division. PMID:26310442

  7. Feature selection for high-dimensional data in astronomy

    NASA Astrophysics Data System (ADS)

    Zheng, Hongwen; Zhang, Yanxia

    With an exponentially increasing amount of astronomical data, the complexity and dimension of astronomical data are likewise growing rapidly. Extracting information from such data becomes a critical and challenging problem. For example, some algorithms can only be employed in the low-dimensional spaces, so feature selection and feature extraction become important topics. Here we describe the difference between feature selection and feature extraction methods, and introduce the taxonomy of feature selection methods as well as the characteristics of each method. We present a case study comparing the performance and computational cost of different feature selection methods. For the filter method, ReliefF and fisher filter are adopted; for the wrapper method, improved CHAID, linear discriminant analysis (LDA), Naive Bayes (NB) and C4.5 are taken as learners. Applied on the sample, the result indicates that from the viewpoints of computational cost the filter method is superior to the wrapper method. Moreover, different learning algorithms combined with appropriate feature selection methods may arrive at better performance.

  8. Micromechanical study of mitotic chromosome structure

    NASA Astrophysics Data System (ADS)

    Marko, John

    2011-03-01

    Our group has developed micromanipulation techniques for study of the highly compacted mitotic form of chromosome found in eukaryote cells during cell division. Each metaphase chromosome contains two duplicate centimeter-long DNA molecules, folded up by proteins into cylindrical structures several microns in length. Native chromosomes display linear and reversible stretching behavior over a wide range of extensions (up to 5x native length for amphibian chromosomes), described by a Young modulus of about 300 Pa. Studies using DNA-cutting and protein-cutting enzymes have revealed that metaphase chromosomes behave as a network of chromatin fibers held together by protein-based isolated crosslinks. Our results are not consistent with the more classical model of loops of chromatin attached to a protein-based structural organizer or ``scaffold". In short, our experiments indicate that metaphase chromosomes can be considered to be ``gels" of chromatin; the stretching modulus of a whole chromosome is consistent with stretching of the chromatin fibers contained within it. Experiments using topoisomerases suggest that topological constraints may play an appreciable role in confining chromatin in the metaphase chromosome. Finally, recent experiments on human chromosomes will be reviewed, including results of experiments where chromosome-folding proteins are specifically depleted using siRNA methods. Supported by NSF-MCB-1022117, DMR-0715099, PHY-0852130, DMR-0520513, NCI 1U54CA143869-01 (NU-PS-OC), and the American Heart Association.

  9. Toward a systems-level view of mitotic checkpoints.

    PubMed

    Ibrahim, Bashar

    2015-03-01

    Reproduction and natural selection are the key elements of life. In order to reproduce, the genetic material must be doubled, separated and placed into two new daughter cells, each containing a complete set of chromosomes and organelles. In mitosis, transition from one process to the next is guided by intricate surveillance mechanisms, known as the mitotic checkpoints. Dis-regulation of cell division through checkpoint malfunction can lead to developmental defects and contribute to the development or progression of tumors. This review approaches two important mitotic checkpoints, the spindle assembly checkpoint (SAC) and the spindle position checkpoint (SPOC). The highly conserved spindle assembly checkpoint (SAC) controls the onset of anaphase by preventing premature segregation of the sister chromatids of the duplicated genome, to the spindle poles. In contrast, the spindle position checkpoint (SPOC), in the budding yeast Saccharomyces cerevisiae, ensures that during asymmetric cell division mitotic exit does not occur until the spindle is properly aligned with the cell polarity axis. Although there are no known homologs, there is indication that functionally similar checkpoints exist also in animal cells. This review can be regarded as an "executable model", which could be easily translated into various quantitative concrete models like Petri nets, ODEs, PDEs, or stochastic particle simulations. It can also function as a base for developing quantitative models explaining the interplay of the various components and proteins controlling mitosis. PMID:25722206

  10. Unusual features of the high light acclimation of Chromera velia.

    PubMed

    Mann, Marcus; Hoppenz, Paul; Jakob, Torsten; Weisheit, Wolfram; Mittag, Maria; Wilhelm, Christian; Goss, Reimund

    2014-11-01

    In the present study, the high light (HL) acclimation of Chromera velia (Chromerida) was studied. HL-grown cells exhibited an increased cell volume and dry weight compared to cells grown at medium light (ML). The chlorophyll (Chl) a-specific absorption spectra ([Formula: see text]) of the HL cells showed an increased absorption efficiency over a wavelength range from 400 to 750 nm, possibly due to differences in the packaging of Chl a molecules. In HL cells, the size of the violaxanthin (V) cycle pigment pool was strongly increased. Despite a higher concentration of de-epoxidized V cycle pigments, non-photochemical quenching (NPQ) of the HL cells was slightly reduced compared to ML cells. The analysis of NPQ recovery during low light (LL) after a short illumination with excess light showed a fast NPQ relaxation and zeaxanthin epoxidation. Purification of the pigment-protein complexes demonstrated that the HL-synthesized V was associated with the chromera light-harvesting complex (CLH). However, the difference absorption spectrum of HL minus ML CLH, together with the 77 K fluorescence excitation spectra, suggested that the additional V was not protein bound but localized in a lipid phase associated with the CLH. The polypeptide analysis of the pigment-protein complexes showed that one out of three known LHCr proteins was associated in higher concentration with photosystem I in the HL cells, whereas in ML cells, it was enriched in the CLH fraction. In conclusion, the acclimation of C. velia to HL illumination shows features that are comparable to those of diatoms, while other characteristics more closely resemble those of higher plants and green algae. PMID:24906888

  11. Cdk-counteracting phosphatases unlock mitotic exit

    PubMed Central

    Queralt, Ethel; Uhlmann, Frank

    2008-01-01

    Entry into mitosis of the eukaryotic cell cycle is driven by rising cyclin-dependent kinase (Cdk) activity. During exit from mitosis, Cdk activity must again decline. Cdk downregulation by itself, however, is not able to guide mitotic exit, if not a phosphatase reverses mitotic Cdk phosphorylation events. In budding yeast, this role is played by the Cdc14 phosphatase. We are gaining an increasingly detailed picture of its regulation during anaphase, and of the way it orchestrates ordered progression through mitosis. Much less is known about protein dephosphorylation during mitotic exit in organisms other than budding yeast, but evidence is now mounting for crucial contributions of regulated phosphatases also in metazoan cells. PMID:18845253

  12. An unmet actin requirement explains the mitotic inhibition of clathrin-mediated endocytosis

    PubMed Central

    Kaur, Satdip; Fielding, Andrew B; Gassner, Gisela; Carter, Nicholas J; Royle, Stephen J

    2014-01-01

    Clathrin-mediated endocytosis (CME) is the major internalisation route for many different receptor types in mammalian cells. CME is shut down during early mitosis, but the mechanism of this inhibition is unclear. In this study, we show that the mitotic shutdown is due to an unmet requirement for actin in CME. In mitotic cells, membrane tension is increased and this invokes a requirement for the actin cytoskeleton to assist the CME machinery to overcome the increased load. However, the actin cytoskeleton is engaged in the formation of a rigid cortex in mitotic cells and is therefore unavailable for deployment. We demonstrate that CME can be ‘restarted’ in mitotic cells despite high membrane tension, by allowing actin to engage in endocytosis. Mitotic phosphorylation of endocytic proteins is maintained in mitotic cells with restored CME, indicating that direct phosphorylation of the CME machinery does not account for shutdown. DOI: http://dx.doi.org/10.7554/eLife.00829.001 PMID:24550251

  13. The Drosophila Microtubule-Associated Protein Mars Stabilizes Mitotic Spindles by Crosslinking Microtubules through Its N-Terminal Region

    PubMed Central

    Zhang, Gang; Beati, Hamze; Nilsson, Jakob; Wodarz, Andreas

    2013-01-01

    Correct segregation of genetic material relies on proper assembly and maintenance of the mitotic spindle. How the highly dynamic microtubules (MTs) are maintained in stable mitotic spindles is a key question to be answered. Motor and non-motor microtubule associated proteins (MAPs) have been reported to stabilize the dynamic spindle through crosslinking adjacent MTs. Mars, a novel MAP, is essential for the early development of Drosophila embryos. Previous studies showed that Mars is required for maintaining an intact mitotic spindle but did not provide a molecular mechanism for this function. Here we show that Mars is able to stabilize the mitotic spindle in vivo. Both in vivo and in vitro data reveal that the N-terminal region of Mars functions in the stabilization of the mitotic spindle by crosslinking adjacent MTs. PMID:23593258

  14. Fluorescent in situ hybridization on mitotic chromosomes of mosquitoes.

    PubMed

    Timoshevskiy, Vladimir A; Sharma, Atashi; Sharakhov, Igor V; Sharakhova, Maria V

    2012-01-01

    Fluorescent in situ hybridization (FISH) is a technique routinely used by many laboratories to determine the chromosomal position of DNA and RNA probes. One important application of this method is the development of high-quality physical maps useful for improving the genome assemblies for various organisms. The natural banding pattern of polytene and mitotic chromosomes provides guidance for the precise ordering and orientation of the genomic supercontigs. Among the three mosquito genera, namely Anopheles, Aedes, and Culex, a well-established chromosome-based mapping technique has been developed only for Anopheles, whose members possess readable polytene chromosomes. As a result of genome mapping efforts, 88% of the An. gambiae genome has been placed to precise chromosome positions. Two other mosquito genera, Aedes and Culex, have poorly polytenized chromosomes because of significant overrepresentation of transposable elements in their genomes. Only 31 and 9% of the genomic supercontings have been assigned without order or orientation to chromosomes of Ae. aegypti and Cx. quinquefasciatus, respectively. Mitotic chromosome preparation for these two species had previously been limited to brain ganglia and cell lines. However, chromosome slides prepared from the brain ganglia of mosquitoes usually contain low numbers of metaphase plates. Also, although a FISH technique has been developed for mitotic chromosomes from a cell line of Ae. aegypti, the accumulation of multiple chromosomal rearrangements in cell line chromosomes makes them useless for genome mapping. Here we describe a simple, robust technique for obtaining high-quality mitotic chromosome preparations from imaginal discs (IDs) of 4th instar larvae which can be used for all three genera of mosquitoes. A standard FISH protocol is optimized for using BAC clones of genomic DNA as a probe on mitotic chromosomes of Ae. aegypti and Cx. quinquefasciatus, and for utilizing an intergenic spacer (IGS) region of ribosomal DNA (rDNA) as a probe on An. gambiae chromosomes. In addition to physical mapping, the developed technique can be applied to population cytogenetics and chromosome taxonomy/systematics of mosquitoes and other insect groups. PMID:23007640

  15. High Resolution Urban Feature Extraction for Global Population Mapping using High Performance Computing

    SciTech Connect

    Vijayaraj, Veeraraghavan; Bright, Eddie A; Bhaduri, Budhendra L

    2007-01-01

    The advent of high spatial resolution satellite imagery like Quick Bird (0.6 meter) and IKONOS (1 meter) has provided a new data source for high resolution urban land cover mapping. Extracting accurate urban regions from high resolution images has many applications and is essential to the population mapping efforts of Oak Ridge National Laboratory's (ORNL) LandScan population distribution program. This paper discusses an automated parallel algorithm that has been implemented on a high performance computing environment to extract urban regions from high resolution images using texture and spectral features

  16. The influence of fixation delay on mitotic activity and flow cytometric cell cycle variables.

    PubMed

    Bergers, E; Jannink, I; van Diest, P I; Cuesta, M A; Meyer, S; van Mourik, J C; Baak, J P

    1997-01-01

    Proliferation variables such as mitotic activity and the percentage of S-phase cells have been shown to be of prognostic value in many tumors, especially in breast cancer. However, some studies reported a decrease in mitotic activity caused by delay in fixation of the tissue. In contrast, other studies showed that the identifiability of mitotic figures decreases after fixation delay, but the total number of mitotic figures and also the percentage of S-phase cells remain unchanged. Most studies have been done on small numbers of experimental tumors, thus introducing the risk of selection bias. The aim of this study was to reinvestigate the influence of fixation delay on mitotic activity and cell cycle variables assessed by flow cytometry in an adequate number of resected human tissues to reach firmer conclusions. Resection specimens of 19 and 21 cases, respectively, for the mitotic activity estimate and the flow cytometric percentage of S-phase calculation were collected directly from the operating theater using lung, breast, and intestinal cancers and normal intestinal mucosa. The tissues were cut in pieces, and from each specimen, pieces were fixed in 4% buffered formaldehyde (for mitosis counting) as well as snap frozen (for flow cytometry) immediately after excision, as well as after a fixation delay of 1, 2, 4, 6, 8, 18, and 24 hours. Moreover, during the fixation delay, one series from each specimen was kept in the refrigerator and the second at room temperature. Thus, a total of 304 (19 X 16) and 336 (21 X 16) specimens were investigated for the mitotic activity estimate and the percentage of S-phase cells calculation, respectively. With regard to the estimation of the mitotic activity, both clear and doubtful mitotic figures were registered separately, obtaining an "uncorrected" and "corrected" (for doubtful mitotic figures) mitotic activity estimate. The percentage of S-phase cells was obtained by cell cycle analysis of flow cytometric DNA-histograms. The results showed that the quality of the material decreased during the fixation delay, as reflected by poorer cellular morphology in the hematoxylin-and-eosin-stained slides, resulting in more difficult identification of mitotic figures and a more time-consuming procedure with regard to the mitosis counts, but not in a worse intraobserver and interobserver reproducibility, which was acceptable. The reduction in quality of the tissues also was shown by the flow cytometric measurements because the coefficient of variation and percentage of debris increased after 4 hours or more of fixation delay. However, the mean values of the "uncorrected" mitotic activity and the "corrected" mitotic activity showed no decreasing trend; neither did the average percentage of S-phase cells. In conclusion, within the time investigated, fixation delay has no clear influence on the proliferation features studied. Because of the decreasing quality of the histological sections, resulting in more difficult identification of mitoses and interpretation of DNA histograms, fixation delay should be kept as short as possible, keeping the tissue at 4 degrees C until fixation. PMID:9013839

  17. Increased mitotic activity as a negative prognostic indicator in pleomorphic xanthoastrocytoma. Case report.

    PubMed

    Macaulay, R J; Jay, V; Hoffman, H J; Becker, L E

    1993-11-01

    Pleomorphic xanthoastrocytoma is a recently characterized neoplasm with a favorable prognosis despite aggressive histological features. The authors report a case of pleomorphic xanthoastrocytoma that recurred 4 years after complete gross resection. The original tumor exhibited histological features characteristic of this neoplasm, but up to 4 mitoses/10 high-power fields were present focally. The recurrent tumor contained small foci of classical pleomorphic xanthoastrocytoma, but consisted predominantly of glioblastoma multiforme. Transitional zones contained nests of glial fibrillary acidic protein (GFAP)-immunopositive cells surrounded by delicate collagenous and reticulin-rich septa. Electron microscopy of the transitional zone showed continuous basal lamina investing cells containing bundles of intermediate filaments. These were GFAP-positive by immunogold electron microscopy, confirming the astrocytic nature of pleomorphic xanthoastrocytoma. This example illustrates the capacity of this tumor to evolve into glioblastoma. The indolent clinical behavior of most pleomorphic xanthoastrocytomas is evident from a literature review, which confirms the prolonged survival of many patients after onset of symptoms. Completeness of excision, subjectively assessed at surgery, did not influence the risk of recurrence or survival up to 10 years after initial resection. Postoperative radiotherapy did not improve survival, but may reduce the probability of recurrence; more studies are needed to corroborate this finding. The data compiled herein support the designation of pleomorphic xanthoastrocytoma as a distinct astrocytic neoplasm with a favorable prognosis. An increased mitotic rate has not previously been correlated with a worse outcome, and should not be used to exclude this diagnosis. However, anaplastic transformation of pleomorphic xanthoastrocytoma confers a much worse prognosis, and this case suggests that increased mitotic activity may be a negative prognostic indicator since it may herald subsequent anaplastic transformation. PMID:8410257

  18. Rapid measurement of mitotic spindle orientation in cultured mammalian cells

    PubMed Central

    Decarreau, Justin; Driver, Jonathan; Asbury, Charles; Wordeman, Linda

    2014-01-01

    Summary Factors that influence the orientation of the mitotic spindle are important for the maintenance of stem cell populations and in cancer development. However, screening for these factors requires rapid quantification of alterations of the angle of the mitotic spindle in cultured cell lines. Here we describe a method to image mitotic cells and rapidly score the angle of the mitotic spindle using a simple MATLAB application to analyze a stack of Z-images. PMID:24633791

  19. Mitotic Stress and Chromosomal Instability in Cancer

    PubMed Central

    Malumbres, Marcos

    2012-01-01

    Cell cycle deregulation is a common motif in human cancer, and multiple therapeutic strategies are aimed to prevent tumor cell proliferation. Whereas most current therapies are designed to arrest cell cycle progression either in G1/S or in mitosis, new proposals include targeting the intrinsic chromosomal instability (CIN, an increased rate of gain or losses of chromosomes during cell division) or aneuploidy (a genomic composition that differs from diploid) that many tumor cells display. Why tumors cells are chromosomally unstable or aneuploid and what are the consequences of these alterations are not completely clear at present. Several mitotic regulators are overexpressed as a consequence of oncogenic alterations, and they are likely to alter the proper regulation of chromosome segregation in cancer cells. In this review, we propose the relevance of TPX2, a mitotic regulator involved in the formation of the mitotic spindle, in oncogene-induced mitotic stress. This protein, as well as its partner Aurora-A, is frequently overexpressed in human cancer, and its deregulation may participate not only in chromosome numeric aberrations but also in other forms of genomic instability in cancer cells. PMID:23634259

  20. APC-dependent proteolysis of the mitotic cyclin Clb2 is essential for mitotic exit.

    PubMed

    Wäsch, Ralph; Cross, Frederick R

    2002-08-01

    Cyclin degradation is central to regulation of the cell cycle. Mitotic exit was proposed to require degradation of the S phase cyclin Clb5 by the anaphase-promoting complex activated by Cdc20 (APC(Cdc20)). Furthermore, Clb5 degradation was thought to be necessary for effective dephosphorylation and activation of the APC regulatory subunit Cdh1 (also known as Hct1) and the cyclin-dependent kinase inhibitor Sic1 by the phosphatase Cdc14, allowing mitotic kinase inactivation and mitotic exit. Here we show, however, that spindle disassembly and cell division occur without significant APC(Cdc20)-mediated Clb5 degradation, as well as in the absence of both Cdh1 and Sic1. We find instead that destruction-box-dependent degradation of the mitotic cyclin Clb2 is essential for mitotic exit. APC(Cdc20) may be required for an essential early phase of Clb2 degradation, and this phase may be sufficient for most aspects of mitotic exit. Cdh1 and Sic1 may be required for further inactivation of Clb2-Cdk1, regulating cell size and the length of G1. PMID:12152084

  1. Identification of Drosophila Mitotic Genes by Combining Co-Expression Analysis and RNA Interference

    PubMed Central

    Somma, Maria Patrizia; Ceprani, Francesca; Bucciarelli, Elisabetta; Naim, Valeria; De Arcangelis, Valeria; Piergentili, Roberto; Palena, Antonella; Ciapponi, Laura; Giansanti, Maria Grazia; Pellacani, Claudia; Petrucci, Romano; Cenci, Giovanni; Vernì, Fiammetta; Fasulo, Barbara; Goldberg, Michael L.; Di Cunto, Ferdinando; Gatti, Maurizio

    2008-01-01

    RNAi screens have, to date, identified many genes required for mitotic divisions of Drosophila tissue culture cells. However, the inventory of such genes remains incomplete. We have combined the powers of bioinformatics and RNAi technology to detect novel mitotic genes. We found that Drosophila genes involved in mitosis tend to be transcriptionally co-expressed. We thus constructed a co-expression–based list of 1,000 genes that are highly enriched in mitotic functions, and we performed RNAi for each of these genes. By limiting the number of genes to be examined, we were able to perform a very detailed phenotypic analysis of RNAi cells. We examined dsRNA-treated cells for possible abnormalities in both chromosome structure and spindle organization. This analysis allowed the identification of 142 mitotic genes, which were subdivided into 18 phenoclusters. Seventy of these genes have not previously been associated with mitotic defects; 30 of them are required for spindle assembly and/or chromosome segregation, and 40 are required to prevent spontaneous chromosome breakage. We note that the latter type of genes has never been detected in previous RNAi screens in any system. Finally, we found that RNAi against genes encoding kinetochore components or highly conserved splicing factors results in identical defects in chromosome segregation, highlighting an unanticipated role of splicing factors in centromere function. These findings indicate that our co-expression–based method for the detection of mitotic functions works remarkably well. We can foresee that elaboration of co-expression lists using genes in the same phenocluster will provide many candidate genes for small-scale RNAi screens aimed at completing the inventory of mitotic proteins. PMID:18797514

  2. Evidence of Selection against Complex Mitotic-Origin Aneuploidy during Preimplantation Development.

    PubMed

    McCoy, Rajiv C; Demko, Zachary P; Ryan, Allison; Banjevic, Milena; Hill, Matthew; Sigurjonsson, Styrmir; Rabinowitz, Matthew; Petrov, Dmitri A

    2015-10-01

    Whole-chromosome imbalances affect over half of early human embryos and are the leading cause of pregnancy loss. While these errors frequently arise in oocyte meiosis, many such whole-chromosome abnormalities affecting cleavage-stage embryos are the result of chromosome missegregation occurring during the initial mitotic cell divisions. The first wave of zygotic genome activation at the 4-8 cell stage results in the arrest of a large proportion of embryos, the vast majority of which contain whole-chromosome abnormalities. Thus, the full spectrum of meiotic and mitotic errors can only be detected by sampling after the initial cell divisions, but prior to this selective filter. Here, we apply 24-chromosome preimplantation genetic screening (PGS) to 28,052 single-cell day-3 blastomere biopsies and 18,387 multi-cell day-5 trophectoderm biopsies from 6,366 in vitro fertilization (IVF) cycles. We precisely characterize the rates and patterns of whole-chromosome abnormalities at each developmental stage and distinguish errors of meiotic and mitotic origin without embryo disaggregation, based on informative chromosomal signatures. We show that mitotic errors frequently involve multiple chromosome losses that are not biased toward maternal or paternal homologs. This outcome is characteristic of spindle abnormalities and chaotic cell division detected in previous studies. In contrast to meiotic errors, our data also show that mitotic errors are not significantly associated with maternal age. PGS patients referred due to previous IVF failure had elevated rates of mitotic error, while patients referred due to recurrent pregnancy loss had elevated rates of meiotic error, controlling for maternal age. These results support the conclusion that mitotic error is the predominant mechanism contributing to pregnancy losses occurring prior to blastocyst formation. This high-resolution view of the full spectrum of whole-chromosome abnormalities affecting early embryos provides insight into the cytogenetic mechanisms underlying their formation and the consequences for human fertility. PMID:26491874

  3. Radmis, a Novel Mitotic Spindle Protein that Functions in Cell Division of Neural Progenitors

    PubMed Central

    Yumoto, Takahito; Nakadate, Kazuhiko; Nakamura, Yuki; Sugitani, Yoshinobu; Sugitani-Yoshida, Reiko; Ueda, Shuichi; Sakakibara, Shin-ichi

    2013-01-01

    Developmental dynamics of neural stem/progenitor cells (NSPCs) are crucial for embryonic and adult neurogenesis, but its regulatory factors are not fully understood. By differential subtractive screening with NSPCs versus their differentiated progenies, we identified the radmis (radial fiber and mitotic spindle)/ckap2l gene, a novel microtubule-associated protein (MAP) enriched in NSPCs. Radmis is a putative substrate for the E3-ubiquitin ligase, anaphase promoting complex/cyclosome (APC/C), and is degraded via the KEN box. Radmis was highly expressed in regions of active neurogenesis throughout life, and its distribution was dynamically regulated during NSPC division. In embryonic and perinatal brains, radmis localized to bipolar mitotic spindles and radial fibers (basal processes) of dividing NSPCs. As central nervous system development proceeded, radmis expression was lost in most brain regions, except for several neurogenic regions. In adult brain, radmis expression persisted in the mitotic spindles of both slowly-dividing stem cells and rapid amplifying progenitors. Overexpression of radmis in vitro induced hyper-stabilization of microtubules, severe defects in mitotic spindle formation, and mitotic arrest. In vivo gain-of-function using in utero electroporation revealed that radmis directed a reduction in NSPC proliferation and a concomitant increase in cell cycle exit, causing a reduction in the Tbr2-positive basal progenitor population and shrinkage of the embryonic subventricular zone. Besides, radmis loss-of-function by shRNAs induced the multipolar mitotic spindle structure, accompanied with the catastrophe of chromosome segregation including the long chromosome bridge between two separating daughter nuclei. These findings uncover the indispensable role of radmis in mitotic spindle formation and cell-cycle progression of NSPCs. PMID:24260314

  4. Arsenite-induced mitotic death involves stress response and is independent of tubulin polymerization

    SciTech Connect

    Taylor, B. Frazier; McNeely, Samuel C.; Miller, Heather L.; States, J. Christopher

    2008-07-15

    Arsenite, a known mitotic disruptor, causes cell cycle arrest and cell death at anaphase. The mechanism causing mitotic arrest is highly disputed. We compared arsenite to the spindle poisons nocodazole and paclitaxel. Immunofluorescence analysis of {alpha}-tubulin in interphase cells demonstrated that, while nocodazole and paclitaxel disrupt microtubule polymerization through destabilization and hyperpolymerization, respectively, microtubules in arsenite-treated cells remain comparable to untreated cells even at supra-therapeutic concentrations. Immunofluorescence analysis of {alpha}-tubulin in mitotic cells showed spindle formation in arsenite- and paclitaxel-treated cells but not in nocodazole-treated cells. Spindle formation in arsenite-treated cells appeared irregular and multi-polar. {gamma}-tubulin staining showed that cells treated with nocodazole and therapeutic concentrations of paclitaxel contained two centrosomes. In contrast, most arsenite-treated mitotic cells contained more than two centrosomes, similar to centrosome abnormalities induced by heat shock. Of the three drugs tested, only arsenite treatment increased expression of the inducible isoform of heat shock protein 70 (HSP70i). HSP70 and HSP90 proteins are intimately involved in centrosome regulation and mitotic spindle formation. HSP90 inhibitor 17-DMAG sensitized cells to arsenite treatment and increased arsenite-induced centrosome abnormalities. Combined treatment of 17-DMAG and arsenite resulted in a supra-additive effect on viability, mitotic arrest, and centrosome abnormalities. Thus, arsenite-induced abnormal centrosome amplification and subsequent mitotic arrest is independent of effects on tubulin polymerization and may be due to specific stresses that are protected against by HSP90 and HSP70.

  5. Evidence of Selection against Complex Mitotic-Origin Aneuploidy during Preimplantation Development

    PubMed Central

    McCoy, Rajiv C.; Demko, Zachary P.; Ryan, Allison; Banjevic, Milena; Hill, Matthew; Sigurjonsson, Styrmir; Rabinowitz, Matthew; Petrov, Dmitri A.

    2015-01-01

    Whole-chromosome imbalances affect over half of early human embryos and are the leading cause of pregnancy loss. While these errors frequently arise in oocyte meiosis, many such whole-chromosome abnormalities affecting cleavage-stage embryos are the result of chromosome missegregation occurring during the initial mitotic cell divisions. The first wave of zygotic genome activation at the 4–8 cell stage results in the arrest of a large proportion of embryos, the vast majority of which contain whole-chromosome abnormalities. Thus, the full spectrum of meiotic and mitotic errors can only be detected by sampling after the initial cell divisions, but prior to this selective filter. Here, we apply 24-chromosome preimplantation genetic screening (PGS) to 28,052 single-cell day-3 blastomere biopsies and 18,387 multi-cell day-5 trophectoderm biopsies from 6,366 in vitro fertilization (IVF) cycles. We precisely characterize the rates and patterns of whole-chromosome abnormalities at each developmental stage and distinguish errors of meiotic and mitotic origin without embryo disaggregation, based on informative chromosomal signatures. We show that mitotic errors frequently involve multiple chromosome losses that are not biased toward maternal or paternal homologs. This outcome is characteristic of spindle abnormalities and chaotic cell division detected in previous studies. In contrast to meiotic errors, our data also show that mitotic errors are not significantly associated with maternal age. PGS patients referred due to previous IVF failure had elevated rates of mitotic error, while patients referred due to recurrent pregnancy loss had elevated rates of meiotic error, controlling for maternal age. These results support the conclusion that mitotic error is the predominant mechanism contributing to pregnancy losses occurring prior to blastocyst formation. This high-resolution view of the full spectrum of whole-chromosome abnormalities affecting early embryos provides insight into the cytogenetic mechanisms underlying their formation and the consequences for human fertility. PMID:26491874

  6. Evidence for a Relatively Random Array of Human Chromosomes on the Mitotic Ring

    PubMed Central

    Allison, David C.; Nestor, Andrea L.

    1999-01-01

    We used fluorescence in situ hybridization (FISH) to study the positions of human chromosomes on the mitotic rings of cultured human lymphocytes, MRC-5 fibroblasts, and CCD-34Lu fibroblasts. The homologous chromosomes of all three cell types had relatively random positions with respect to each other on the mitotic rings of prometaphase rosettes and anaphase cells. Also, the positions of the X and Y chromosomes, colocalized with the somatic homologues in male cells, were highly variable from one mitotic ring to another. Although random chromosomal positions were found in different pairs of CCD-34Lu and MRC-5 late-anaphases, the separations between the same homologous chromosomes in paired late-anaphase and telophase chromosomal masses were highly correlated. Thus, although some loose spatial associations of chromosomes secondary to interphase positioning may exist on the mitotic rings of some cells, a fixed order of human chromosomes and/or a rigorous separation of homologous chromosomes on the mitotic ring are not necessary for normal mitosis. Furthermore, the relative chromosomal positions on each individual metaphase plate are most likely carried through anaphase into telophase. PMID:10189364

  7. Mitotic inhibition of clathrin-mediated endocytosis

    PubMed Central

    Fielding, Andrew B.; Royle, Stephen J.

    2014-01-01

    Endocytosis and mitosis are fundamental processes in a cell’s life. Nearly fifty years of research suggest that these processes are linked and that endocytosis is shut down as cells undergo the early stages of mitosis. Precisely how this occurs at a molecular level is an open question. In this review, we summarize the early work characterizing the inhibition of clathrin-mediated endocytosis and discuss recent challenges to this established concept. We also set out four proposed mechanisms for the inhibition: mitotic phosphorylation of endocytic proteins, altered membrane tension, moonlighting of endocytic proteins and a mitotic spindle-dependent mechanism. Finally, we speculate the functional consequences of endocytic shutdown during mitosis and where an understanding of the mechanism of inhibition will lead us in the future. PMID:23307073

  8. Automatic microscopy for mitotic cell location.

    NASA Technical Reports Server (NTRS)

    Herron, J.; Ranshaw, R.; Castle, J.; Wald, N.

    1972-01-01

    Advances are reported in the development of an automatic microscope with which to locate hematologic or other cells in mitosis for subsequent chromosome analysis. The system under development is designed to perform the functions of: slide scanning to locate metaphase cells; conversion of images of selected cells into binary form; and on-line computer analysis of the digitized image for significant cytogenetic data. Cell detection criteria are evaluated using a test sample of 100 mitotic cells and 100 artifacts.

  9. Emission features in the spectrum of NGC 7027 near 3. 3 microns at very high resolution

    SciTech Connect

    Lowe, R.P.; Moorhead, J.M.; Wehlau, W.H.; Maillard, J.P. CNRS, Institut d'Astrophysique, Paris )

    1991-02-01

    A very high resolution spectrum is presented of the planetary nebula NGC 7027 over a 200/cm interval centered at 2950/cm, and the features found are described: (1) nebular continuum, (2) atomic recombination lines of H and He II, and (3) three broader emission features of uncertain origin. For the latter the first evidence is presented that the 3.46 micron feature and possibly the 3.40 micron feature are resolvable into a sequence of narrower features. The interpretation of the broader features is discussed in terms of the hypothesis of identification with emission by polycyclic aromatic hydrocarbons. 18 refs.

  10. Practical features of illumination with high pressure sodium lamps

    SciTech Connect

    Corth, R.

    1983-06-01

    A number of concerns raised about the health effects of high pressure sodium lamps (HPS) are discussed. The notion of a ''natural'' human photic environment based on sunlight is disputed. Humans are better adapted to the ''greenish'' spectral composition of forest light than to direct sunlight. It is ironic that the artificial light source which has received the most disapproval, cool white flourescent lamp, has a spectral composition similar to that of forest light. HPS is also available in a full range of colors. Some successful examples of HPS--from North Division High School, in Milwaukee, Wisconsin, to museum exhibits at National Geographic in Washington--are listed.

  11. Mitotic spindle studied using picosecond laser scissors

    NASA Astrophysics Data System (ADS)

    Baker, N. M.; Botvinick, E. L.; Shi, Linda; Berns, M. B.; Wu, George

    2006-08-01

    In previous studies we have shown that the second harmonic 532 nm, from a picosecond frequency doubled Nd:YAG laser, can cleanly and selectively disrupt spindle fiber microtubules in live cells (Botvinick et al 2004, Biophys. J. 87:4303-4212). In the present study we have ablated different locations and amounts of the metaphase mitotic spindle, and followed the cells in order to observe the fate of the irradiated spindle and the ability of the cell to continue through mitosis. Cells of the rat kangaroo line (PTK2) were stably transfected by ECFP-tubulin and, using fluorescent microscopy and the automated RoboLase microscope, (Botvinick and Berns, 2005, Micros. Res. Tech. 68:65-74) brightly fluorescent individual cells in metaphase were irradiated with 0.2447 nJ/micropulse corresponding to an irradiance of 1.4496*10^7 J/(ps*cm^2) . Upon irradiation the exposed part of the mitotic spindle immediately lost fluorescence and the following events were observed in the cells over time: (1) immediate contraction of the spindle pole towards the cut, (2) recovery of connection between pole and cut microtubule, (3) completion of mitosis. This system should be very useful in studying internal cellular dynamics of the mitotic spindle.

  12. DEK over-expression promotes mitotic defects and micronucleus formation

    PubMed Central

    Matrka, Marie C; Hennigan, Robert F; Kappes, Ferdinand; DeLay, Monica L; Lambert, Paul F; Aronow, Bruce J; Wells, Susanne I

    2015-01-01

    The DEK gene encodes a nuclear protein that binds chromatin and is involved in various fundamental nuclear processes including transcription, RNA splicing, DNA replication and DNA repair. Several cancer types characteristically over-express DEK at the earliest stages of transformation. In order to explore relevant mechanisms whereby DEK supports oncogenicity, we utilized cancer databases to identify gene transcripts whose expression patterns are tightly correlated with that of DEK. We identified an enrichment of genes involved in mitosis and thus investigated the regulation and possible function of DEK in cell division. Immunofluorescence analyses revealed that DEK dissociates from DNA in early prophase and re-associates with DNA during telophase in human keratinocytes. Mitotic cell populations displayed a sharp reduction in DEK protein levels compared to the corresponding interphase population, suggesting DEK may be degraded or otherwise removed from the cell prior to mitosis. Interestingly, DEK overexpression stimulated its own aberrant association with chromatin throughout mitosis. Furthermore, DEK co-localized with anaphase bridges, chromosome fragments, and micronuclei, suggesting a specific association with mitotically defective chromosomes. We found that DEK over-expression in both non-transformed and transformed cells is sufficient to stimulate micronucleus formation. These data support a model wherein normal chromosomal clearance of DEK is required for maintenance of high fidelity cell division and chromosomal integrity. Therefore, the overexpression of DEK and its incomplete removal from mitotic chromosomes promotes genomic instability through the generation of genetically abnormal daughter cells. Consequently, DEK over-expression may be involved in the initial steps of developing oncogenic mutations in cells leading to cancer initiation PMID:25945971

  13. STEM High School Communities: Common and Differing Features

    ERIC Educational Resources Information Center

    Tofel-Grehl, Colby; Callahan, Carolyn M.

    2014-01-01

    Using observations and interviews, the researchers explore the experiences and perspectives of students, teachers, and administrators at six specialized high schools with a focus on science, technology, engineering, and mathematics (STEM) as they pertain to the practices and structures affecting student outcomes. Four themes were found to be…

  14. Incremental slow feature analysis: adaptive low-complexity slow feature updating from high-dimensional input streams.

    PubMed

    Kompella, Varun Raj; Luciw, Matthew; Schmidhuber, Jrgen

    2012-11-01

    We introduce here an incremental version of slow feature analysis (IncSFA), combining candid covariance-free incremental principal components analysis (CCIPCA) and covariance-free incremental minor components analysis (CIMCA). IncSFA's feature updating complexity is linear with respect to the input dimensionality, while batch SFA's (BSFA) updating complexity is cubic. IncSFA does not need to store, or even compute, any covariance matrices. The drawback to IncSFA is data efficiency: it does not use each data point as effectively as BSFA. But IncSFA allows SFA to be tractably applied, with just a few parameters, directly on high-dimensional input streams (e.g., visual input of an autonomous agent), while BSFA has to resort to hierarchical receptive-field-based architectures when the input dimension is too high. Further, IncSFA's updates have simple Hebbian and anti-Hebbian forms, extending the biological plausibility of SFA. Experimental results show IncSFA learns the same set of features as BSFA and can handle a few cases where BSFA fails. PMID:22845826

  15. Dietary flavonoid fisetin induces a forced exit from mitosis by targeting the mitotic spindle checkpoint

    PubMed Central

    Salmela, Anna-Leena; Pouwels, Jeroen; Varis, Asta; Kukkonen, Anu M.; Toivonen, Pauliina; Halonen, Pasi K.; Perälä, Merja; Kallioniemi, Olli; Gorbsky, Gary J.; Kallio, Marko J.

    2009-01-01

    Fisetin is a natural flavonol present in edible vegetables, fruits and wine at 2–160 μg/g concentrations and an ingredient in nutritional supplements with much higher concentrations. The compound has been reported to exert anticarcinogenic effects as well as antioxidant and anti-inflammatory activity via its ability to act as an inhibitor of cell proliferation and free radical scavenger, respectively. Our cell-based high-throughput screen for small molecules that override chemically induced mitotic arrest identified fisetin as an antimitotic compound. Fisetin rapidly compromised microtubule drug-induced mitotic block in a proteasome-dependent manner in several human cell lines. Moreover, in unperturbed human cancer cells fisetin caused premature initiation of chromosome segregation and exit from mitosis without normal cytokinesis. To understand the molecular mechanism behind these mitotic errors, we analyzed the consequences of fisetin treatment on the localization and phoshorylation of several mitotic proteins. Aurora B, Bub1, BubR1 and Cenp-F rapidly lost their kinetochore/centromere localization and others became dephosphorylated upon addition of fisetin to the culture medium. Finally, we identified Aurora B kinase as a novel direct target of fisetin. The activity of Aurora B was significantly reduced by fisetin in vitro and in cells, an effect that can explain the observed forced mitotic exit, failure of cytokinesis and decreased cell viability. In conclusion, our data propose that fisetin perturbs spindle checkpoint signaling, which may contribute to the antiproliferative effects of the compound. PMID:19395653

  16. Feature correlation hypergraph: exploiting high-order potentials for multimodal recognition.

    PubMed

    Zhang, Luming; Gao, Yue; Hong, Chaoqun; Feng, Yinfu; Zhu, Jianke; Cai, Deng

    2014-08-01

    In computer vision and multimedia analysis, it is common to use multiple features (or multimodal features) to represent an object. For example, to well characterize a natural scene image, we typically extract a set of visual features to represent its color, texture, and shape. However, it is challenging to integrate multimodal features optimally. Since they are usually high-order correlated, e.g., the histogram of gradient (HOG), bag of scale invariant feature transform descriptors, and wavelets are closely related because they collaboratively reflect the image texture. Nevertheless, the existing algorithms fail to capture the high-order correlation among multimodal features. To solve this problem, we present a new multimodal feature integration framework. Particularly, we first define a new measure to capture the high-order correlation among the multimodal features, which can be deemed as a direct extension of the previous binary correlation. Therefore, we construct a feature correlation hypergraph (FCH) to model the high-order relations among multimodal features. Finally, a clustering algorithm is performed on FCH to group the original multimodal features into a set of partitions. Moreover, a multiclass boosting strategy is developed to obtain a strong classifier by combining the weak classifiers learned from each partition. The experimental results on seven popular datasets show the effectiveness of our approach. PMID:24184790

  17. Micalastic high-voltage insulation: Design features and experience

    NASA Astrophysics Data System (ADS)

    Wichmann, A.

    1981-12-01

    High-quality mica, carefully selected epoxy resins and a well-matched vacuum/pressure impregnation process determine the characteristics of the MICALASTIC insulation for large turbine-generators. Logical development and process manufacturing quality control have led to an insulation system of high quality and operating reliability. The first winding of a turbine-generator being impregnated and cured under vacuum with solvent-free synthetic resin in 1958 was designed for 10.5 kV rated voltage. Ever since, Siemens AG and Kraftwerk Union AG have used this type of insulation for all direct-cooled windings and also for an increasing number of indirect-cooled windings. At present, 240 turbine-generators with a total of more than 115,000 MVA output have been built. Since 1960, this insulation system has been registered for Siemens AG under the trade name MICALASTIC. The stator windings of the largest, single-shaft generators to date, rated 1560 MVA, 27 kV, has been built with MICALASTIC insulation.

  18. Pair normalized channel feature and statistics-based learning for high-performance pedestrian detection

    NASA Astrophysics Data System (ADS)

    Zeng, Bobo; Wang, Guijin; Ruan, Zhiwei; Lin, Xinggang; Meng, Long

    2012-07-01

    High-performance pedestrian detection with good accuracy and fast speed is an important yet challenging task in computer vision. We design a novel feature named pair normalized channel feature (PNCF), which simultaneously combines and normalizes two channel features in image channels, achieving a highly discriminative power and computational efficiency. PNCF applies to both gradient channels and color channels so that shape and appearance information are described and integrated in the same feature. To efficiently explore the formidably large PNCF feature space, we propose a statistics-based feature learning method to select a small number of potentially discriminative candidate features, which are fed into the boosting algorithm. In addition, channel compression and a hybrid pyramid are employed to speed up the multiscale detection. Experiments illustrate the effectiveness of PNCF and its learning method. Our proposed detector outperforms the state-of-the-art on several benchmark datasets in both detection accuracy and efficiency.

  19. Highly featured amorphous silicon nanorod arrays for high-performance lithium-ion batteries

    SciTech Connect

    Soleimani-Amiri, Samaneh; Safiabadi Tali, Seied Ali; Azimi, Soheil; Sanaee, Zeinab; Mohajerzadeh, Shamsoddin

    2014-11-10

    High aspect-ratio vertical structures of amorphous silicon have been realized using hydrogen-assisted low-density plasma reactive ion etching. Amorphous silicon layers with the thicknesses ranging from 0.5 to 10 μm were deposited using radio frequency plasma enhanced chemical vapor deposition technique. Standard photolithography and nanosphere colloidal lithography were employed to realize ultra-small features of the amorphous silicon. The performance of the patterned amorphous silicon structures as a lithium-ion battery electrode was investigated using galvanostatic charge-discharge tests. The patterned structures showed a superior Li-ion battery performance compared to planar amorphous silicon. Such structures are suitable for high current Li-ion battery applications such as electric vehicles.

  20. Differential Mitotic Stability of Yeast Disomes Derived from Triploid Meiosis

    PubMed Central

    Campbell, Douglas; Doctor, John S.; Feuersanger, Jeane H.; Doolittle, Mark M.

    1981-01-01

    The frequencies of recovered disomy among the meiotic segregants of yeast (Saccharomyces cerevisiae) triploids were assessed under conditions in which all 17 yeast chromosomes were monitored simultaneously. The studies employed inbred triploids, in which all homologous centromeres were identical by descent, and single haploid testers carrying genetic markers for all 17 linkage groups. The principal results include: (1) Ascospores from triploid meiosis germinate at frequencies comparable to those from normal diploids, but most fail to produce visible colonies due to the growth-retarding effects of high multiple disomy. (2) The probability of disome formation during triploid meiosis is the same for all chromosomes; disomy for any given chromosome does not exclude simultaneous disomy for any other chromosome. (3) The 17 yeast chromosomes fall into three frequency classes in terms of disome recovery. The results support the idea that multiply disomic meiotic segregants of the triploid experience repeated, nonrandom, post-germination mitotic chromosome losses (N+1→N) and that the observed variations in individual disome recovery are wholly attributable to inherent differences in disome mitotic stability. PMID:7035289

  1. Correlation of histopathologic features of ductal carcinoma in situ of the breast with the oncotype DX DCIS score.

    PubMed

    Knopfelmacher, Adriana; Fox, Jana; Lo, Yungtai; Shapiro, Nella; Fineberg, Susan

    2015-09-01

    The Oncotype DX Breast Cancer Assay for ductal carcinoma in situ is used to determine local recurrence risk in patients with ductal carcinoma in situ. The results help select patients with low-risk ductal carcinoma in situ who could forgo radiation therapy after conservative surgery. The genes assessed include five proliferation genes, progesterone receptor (PR), and GSTM-1. Our objective was to determine if PR, mitotic counting, or any other pathologic feature of ductal carcinoma in situ could predict the Oncotype DX DCIS Score. We identified 46 cases of ductal carcinoma in situ with a Oncotype DX DCIS Score. In addition to information obtained from routine pathology, we counted mitotic figures in the ductal carcinoma in situ and noted presence of dense chronic inflammatory infiltrate surrounding ductal carcinoma in situ. We found that PR ≥ 90% (P = 0.004), mitotic count ≤ 1 (P = 0.045), estrogen receptor ≥ 90% (P = 0.046), and low nuclear grade (P < 0.0001) were associated with a low score. Dense chronic inflammation surrounding ductal carcinoma in situ was associated with a high score (P = 0.034).All 13 cases with PR ≥ 90%, ≤ 1 mitotic figure and absence of dense chronic inflammation around ductal carcinoma in situ had a low score (100% specificity). A low score was not observed in any case with at least two of the following--negative PR, >1 mitotic figure, and/or presence of dense chronic inflammation around ductal carcinoma in situ (100% specificity). Our study suggests using a combination of PR (≥ 90% vs negative) with mitotic count in ductal carcinoma in situ (≤ 1 vs >1) and dense chronic inflammation around ductal carcinoma in situ one could predict the Oncotype DX DCIS score. Mitotic counting and evaluation of immune response might provide prognostic information in ductal carcinoma in situ. PMID:26111975

  2. Global Phosphoproteomic Mapping of Early Mitotic Exit in Human Cells Identifies Novel Substrate Dephosphorylation Motifs.

    PubMed

    McCloy, Rachael A; Parker, Benjamin L; Rogers, Samuel; Chaudhuri, Rima; Gayevskiy, Velimir; Hoffman, Nolan J; Ali, Naveid; Watkins, D Neil; Daly, Roger J; James, David E; Lorca, Thierry; Castro, Anna; Burgess, Andrew

    2015-08-01

    Entry into mitosis is driven by the coordinated phosphorylation of thousands of proteins. For the cell to complete mitosis and divide into two identical daughter cells it must regulate dephosphorylation of these proteins in a highly ordered, temporal manner. There is currently a lack of a complete understanding of the phosphorylation changes that occur during the initial stages of mitotic exit in human cells. Therefore, we performed a large unbiased, global analysis to map the very first dephosphorylation events that occur as cells exit mitosis. We identified and quantified the modification of >16,000 phosphosites on >3300 unique proteins during early mitotic exit, providing up to eightfold greater resolution than previous studies. The data have been deposited to the ProteomeXchange with identifier PXD001559. Only a small fraction (∼ 10%) of phosphorylation sites were dephosphorylated during early mitotic exit and these occurred on proteins involved in critical early exit events, including organization of the mitotic spindle, the spindle assembly checkpoint, and reformation of the nuclear envelope. Surprisingly this enrichment was observed across all kinase consensus motifs, indicating that it is independent of the upstream phosphorylating kinase. Therefore, dephosphorylation of these sites is likely determined by the specificity of phosphatase/s rather than the activity of kinase/s. Dephosphorylation was significantly affected by the amino acids at and surrounding the phosphorylation site, with several unique evolutionarily conserved amino acids correlating strongly with phosphorylation status. These data provide a potential mechanism for the specificity of phosphatases, and how they co-ordinate the ordered events of mitotic exit. In summary, our results provide a global overview of the phosphorylation changes that occur during the very first stages of mitotic exit, providing novel mechanistic insight into how phosphatase/s specifically regulate this critical transition. PMID:26055452

  3. The effects of testosterone and oestrogen on gonadectomised and intact male rat anterior pituitary mitotic and apoptotic activity.

    PubMed

    Nolan, L A; Levy, A

    2006-03-01

    We have used a direct, non-immunochemical and highly accurate method to quantify the effects of testosterone and oestrogen on mitotic and apoptotic activity in the young, male rat anterior pituitary in vivo. Surgical gonadectomy resulted in a 3-fold increase in mitotic activity by the fourth post-operative day, which returned gradually to levels seen in intact animals over the subsequent 3-4 weeks. Both a single dose of Sustanon, a mixture of long-acting testosterone esters in arachis oil, and the same dose divided over 7 days (starting 6 days after gonadectomy), initially suppressed mitotic activity to levels seen in intact animals, but was associated after 48-96 h with a wave of increased mitotic activity. The latter was blocked by co-administration of Sustanon with the non-steroidal aromatase inhibitor letrozole and was not seen when the non-aromatisable androgen dihydrotestosterone was substituted for Sustanon. Oestrogen alone in gonadectomised and intact rats produced a marked increase in mitosis as expected. With the exception of a transient increase in response to a single high-dose injection of Sustanon in gonadectomised animals, apoptotic activity was unaffected by all of the above. This study suggests that pituitary mitotic activity is tonically inhibited by gonadal hormone production (at least in the short term) in adult male rats. The study also suggests that supraphysiological testosterone treatment -- while unable to reduce anterior pituitary mitotic activity in untreated, intact animals --suppresses the early increase in mitotic activity induced by gonadectomy. Oestrogen, either exogenous or generated locally by aromatisation, stimulates anterior pituitary mitotic activity in a time-dependent manner. PMID:16522719

  4. Induction of mitotic cell division distrubances and mitotic arrest by pyrethroids in V79 cell cultures.

    PubMed

    Hadnagy, W; Seemayer, N H; Kühn, K H; Leng, G; Idel, H

    1999-06-30

    Five pyrethroids (fenvalerate, deltamethrin, cypermethrin, permethrin, cyfluthrin) differing in their chemical purity were investigated on their cytotoxic effects, especially on their ability to induce mitotic cell division disturbances using Chinese hamster lung cells of line V79. The colony forming ability (CFA) resulted in distinct differences of the cytotoxic effect of the tested pyrethroids, whereby permethrin was found to be most toxic. With the exception of fenvalerate all tested pyrethroids gave rise to inhibition of cell cycle progression as shown by G2/M-arrest of synchronized V79 cells by flow cytometry as well as by the increase of the mitotic index as evaluated by light microscopy. The mitotic arresting activity could be attributed to the occurrence of abnormal mitotic figures such as initial and full C-metaphases. The results however indicate, that pyrethroids per se do not contribute to the cytotoxic effects but that other factors such as chemical impurities, source as well as manufacturing process and isomer composition may be responsible for the observed cytotoxic effects. PMID:10414784

  5. Non-iridescent Transmissive Structural Color Filter Featuring Highly Efficient Transmission and High Excitation Purity

    PubMed Central

    Shrestha, Vivek Raj; Lee, Sang-Shin; Kim, Eun-Soo; Choi, Duk-Yong

    2014-01-01

    Nanostructure based color filtering has been considered an attractive replacement for current colorant pigmentation in the display technologies, in view of its increased efficiencies, ease of fabrication and eco-friendliness. For such structural filtering, iridescence relevant to its angular dependency, which poses a detrimental barrier to the practical development of high performance display and sensing devices, should be mitigated. We report on a non-iridescent transmissive structural color filter, fabricated in a large area of 76.2 × 25.4 mm2, taking advantage of a stack of three etalon resonators in dielectric films based on a high-index cavity in amorphous silicon. The proposed filter features a high transmission above 80%, a high excitation purity of 0.93 and non-iridescence over a range of 160°, exhibiting no significant change in the center wavelength, dominant wavelength and excitation purity, which implies no change in hue and saturation of the output color. The proposed structure may find its potential applications to large-scale display and imaging sensor systems. PMID:24815530

  6. Quantitative thresholds for mitotic counts in histologic grading: confirmation in nonfrozen samples of invasive ductal breast cancer.

    PubMed

    Kronqvist, P; Kuopio, T; Collan, Y

    2000-04-01

    Increasing evidence in the medical literature suggests that freezing a sample before fixation causes changes in the histologically observed mitotic activity. In a recent study we determined quantitative thresholds for mitotic counts in invasive ductal breast cancer in both nonfrozen and frozen formalin-fixed specimens. Survival- and recurrence-based analyses of this study material indicated grading thresholds of 17 and 32 mitoses/mm(2) for standardized mitotic index (SMI) and 13 and 35 mitoses/10 high-power fields for mitotic activity index (MAI). The purpose of the present study is to confirm and adjust the introduced thresholds in only nonfrozen formalin-fixed samples. The SMI and MAI in 202 cases of nonfrozen formalin-fixed samples were analyzed to determine optimal cutpoints for prognostication of invasive breast cancer on the basis of mitotic activity. The SMI thresholds were identical in both the present nonfrozen specimens and the previous combined specimens. The optimal MAI thresholds in the nonfrozen material changed to 11 and 37 mitoses/10 high-power field. The confirmation and adjustment of the mitotic thresholds improved the prognostic significance of the method in the nonfrozen material, which will contribute to the clinical applicability of a morphometric grading system. PMID:10760318

  7. Post-slippage multinucleation renders cytotoxic variation in anti-mitotic drugs that target the microtubules or mitotic spindle

    PubMed Central

    Zhu, Yanting; Zhou, Yuan; Shi, Jue

    2014-01-01

    One common cancer chemotherapeutic strategy is to perturb cell division with anti-mitotic drugs. Paclitaxel, the classic microtubule-targeting anti-mitotic drug, so far still outperforms the newer, more spindle-specific anti-mitotics in the clinic, but the underlying cellular mechanism is poorly understood. In this study we identified post-slippage multinucleation, which triggered extensive DNA damage and apoptosis after drug-induced mitotic slippage, contributes to the extra cytotoxicity of paclitaxel in comparison to the spindle-targeting drug, Kinesin-5 inhibitor. Based on quantitative single-cell microscopy assays, we showed that attenuation of the degree of post-slippage multinucleation significantly reduced DNA damage and apoptosis in response to paclitaxel, and that post-slippage apoptosis was likely mediated by the p53-dependent DNA damage response pathway. Paclitaxel appeared to act as a double-edge sword, capable of killing proliferating cancer cells both during mitotic arrest and after mitotic slippage by inducing DNA damage. Our results thus suggest that to predict drug response to paclitaxel and anti-mitotics in general, 2 distinct sets of bio-markers, which regulate mitotic and post-slippage cytotoxicity, respectively, may need to be considered. Our findings provide important new insight not only for elucidating the cytotoxic mechanisms of paclitaxel, but also for understanding the variable efficacy of different anti-mitotic chemotherapeutics. PMID:24694730

  8. Mitotic recombination of chromosome 17 in astrocytomas

    SciTech Connect

    James, C.D.; Carlbom, E.; Nordenskjold, M.; Collins, V.P.; Cavenee, W.K. )

    1989-04-01

    Allelic combinations at seven loci on human chromosome 17 defined by restriction fragment length polymorphisms were determined in tumor and normal tissues from 35 patients with gliomas. Loss of constitutional heterozygosity at one or more of these loci was observed in 8 of the 24 tumors displaying astrocytic differentiation and in the single primitive neuroectodermal tumor examined. The astrocytomas showing these losses included examples of each adult malignancy grade of the disease, including glioblastoma (malignancy grade IV), and seven of them demonstrated concurrent maintenance of heterozygosity for at least one chromosome 17 locus. Determination of allele dosage together with the genotypic data indicated that the tumor chromosomes 17 were derived by mitotic recombination in 7 of the 9 cases with shared homozygosity of the region 17p11.2-ptr in all cases. In contrast, tumors of oligodendrocytic, ependymal, or mixed cellular differentiation did not exhibit loss of alleles at any of the loci examined. These data suggest that the somatic attainment of homozygosity for loci on chromosome 17p is frequently associated with the oncogenesis of central nervous system tumors, particularly those showing solely astrocytic differentiation, and that mitotic recombination mapping is a useful approach towards the subregional localization of a locus whose rearrangement is involved in this disease.

  9. Induction of mitotic aneuploidy in lower eukaryotes

    SciTech Connect

    Kappas, A.

    1993-12-31

    Genetic tests for induction of mitotic aneuploidy in lower eukarotes used mainly the fungal systems of Aspergillus nidulans and Saccharomyces cerevisiae. There are several differences between the two systems such as the greater tolerance for aneuploidy and the fertility of triploids in S. cerevisiae, the stability of diploids and the selective advantage of haploids over diploids in Aspergillus and the mycelial growth of Aspergillus. On the other hand several similarities also exist between the two systems such as the general instability and varying growth rate of disomics and the random loss of extra chromosomes which produces more competitive types or the most frequent recovery of certain specific aneuploids. In using lower eukaryotes as test systems for the identification of aneugens several points should be considered which concern the relevance of such systems to higher organisms, the ability to identify primary aneuploidy and distinguish this from events, such as chromosomal breaks, which lead to secondary aneuploidy and the ability to obtain repeatable results. Within the framework of an EEC comparative study for evaluating assays for aneuploidy, a number of chemicals were assayed in A. nidulans for mitotic instability due to malsegregation of chromosomes at cell division.

  10. A mitotically active, cellular tumor stroma and/or inflammatory cells associated with tumor cells may contribute to intermediate or high Oncotype DX Recurrence Scores in low-grade invasive breast carcinomas.

    PubMed

    Acs, Geza; Esposito, Nicole N; Kiluk, John; Loftus, Loretta; Laronga, Christine

    2012-04-01

    Oncotype DX is an RT-PCR-based 21-gene assay validated to provide prognostic and predictive information in the form of a Recurrence Score in patients with estrogen receptor-positive, lymph node-negative breast cancer. Although the Recurrence Score was shown to correlate with several histopathological tumor features, there is a significant proportion of cases showing an apparent discrepancy between Recurrence Score and risk estimates based on the traditional clinicopathological tumor features. In this study, we tested whether a proliferating, cellular stroma and/or admixed inflammatory cells may result in an artificially increased Recurrence Score in low-grade invasive breast cancers. We analyzed the histopathological features in 141 low-grade invasive breast carcinomas, including 41 special type (tubular, cribriform and mucinous) carcinomas, with available Recurrence Score. The tumor stroma was evaluated for increased cellularity and presence of inflammatory cells. Double immunohistochemical stains for pancytokeratin and Ki-67 was performed to assess the cell proliferation in tumor vs stromal/inflammatory cells. The clinicopathological features of tumors with Recurrence Score <18 (low risk) were compared with those with Recurrence Score ≥18 (intermediate/high risk). Carcinomas associated with Recurrence Score ≥18 showed lower progesterone receptor immunoreactivity, increased stromal cellularity and presence of inflammatory cells associated with the tumor. Double immunohistochemical stains showed significantly increased proliferation in stromal/inflammatory cells compared with carcinoma cells in cases associated with Recurrence Score ≥18. A Ki-67-positive stromal/tumor cells ratio of >1 predicted Recurrence Score ≥18 with an area under the curve of 0.8967 on receiver operator curve analysis (P<0.0001). Our results suggest that the presence of increased stromal cellularity and/or associated inflammatory cells in low-grade invasive breast carcinomas may contribute to an apparently increased risk of recurrence according to Oncotype DX Recurrence Score. Careful assessment and correlation with histopathological features in such cases may help in determining the appropriate patient management. PMID:22173289

  11. The transforming acidic coiled coil 3 protein is essential for spindle-dependent chromosome alignment and mitotic survival.

    PubMed

    Schneider, Leonid; Essmann, Frank; Kletke, Anja; Rio, Paula; Hanenberg, Helmut; Wetzel, Wiebke; Schulze-Osthoff, Klaus; Nürnberg, Bernd; Piekorz, Roland P

    2007-10-01

    Cancer-associated centrosomal transforming acidic coiled coil (TACC) proteins are involved in mitotic spindle function. By employing gene targeting, we have recently described a nonredundant and essential role of TACC3 in regulating cell proliferation. In this study, we used an inducible RNA interference approach to characterize the molecular function of TACC3 and its role in mitotic progression and cell survival. Our data demonstrate that a TACC3 knockdown arrests G(1) checkpoint-compromised HeLa cells prior to anaphase with aberrant spindle morphology and severely misaligned chromosomes. Interestingly, TACC3-depleted cells fail to accumulate the mitotic kinase Aurora B and the checkpoint protein BubR1 to normal levels at kinetochores. Moreover, localization of the structural protein Ndc80 at outer kinetochores is reduced, indicating a defective kinetochore-microtubule attachment in TACC3-deficient cells. As a consequence of prolonged TACC3 depletion, cells undergo caspase-dependent cell death that relies on a spindle checkpoint-dependent mitotic arrest. TACC3 knockdown cells that escape from this arrest by mitotic slippage become highly polyploid and accumulate supernumerary centrosomes. Similarly, deficiency of the post-mitotic cell cycle inhibitor p21(WAF) exacerbates the effects of TACC3 depletion. Our findings therefore point to an essential role of TACC3 in spindle assembly and cellular survival and identify TACC3 as a potential therapeutic target in cancer cells. PMID:17675670

  12. Random mitotic activities across human embryonic stem cell colonies.

    SciTech Connect

    Jin, Q.; Duggan, R.; Dasa, S.; Li, F.; Chen, L.

    2010-08-01

    A systemic and quantitative study was performed to examine whether different levels of mitotic activities, assessed by the percentage of S-phase cells at any given time point, existed at different physical regions of human embryonic stem (hES) cell colonies at 2, 4, 6 days after cell passaging. Mitotically active cells were identified by the positive incorporation of 5-bromo-2-deoxyuridine (BrdU) within their newly synthesized DNA. Our data indicated that mitotically active cells were often distributed as clusters randomly across the colonies within the examined growth period, presumably resulting from local deposition of newly divided cells. This latter notion was further demonstrated by the confined growth of enhanced green florescence protein (EGFP) expressing cells amongst non-GFP expressing cells. Furthermore, the overall percentage of mitotically active cells remained constantly at about 50% throughout the 6-day culture period, indicating mitotic activities of hES cell cultures were time-independent under current growth conditions.

  13. A new role for Rab GTPases during early mitotic stages.

    PubMed

    Das, Sanchaita; Hehnly, Heidi; Doxsey, Stephen

    2014-01-01

    A recent study revealed new roles for the Rab11 GTPase during mitosis. Rab11 is involved in recycling endosome localization to mitotic spindle poles via dynein-mediated transport. This process is in contrast to Golgi membranes, which disperse in mitosis and do not appear to directly contribute to mitotic functions. Rab11-depletion prevents recycling endosome organization at spindle poles, delays mitotic progression, and disrupts spindle pole protein recruitment, astral microtubule organization, and mitotic spindle orientation. However, Rab11 is not the only endocytic and/or trafficking protein that regulates mitotic progression. Clathrin and two small GTPases (Rab6A', Rab5) play key roles in spindle organization and function. In this commentary, we discuss the roles of all these canonical endocytic and membrane trafficking proteins during mitosis and speculate on possible cross-communication between them and their molecular pathways that ensure faithful progression through mitosis. PMID:24921241

  14. A new role for Rab GTPases during early mitotic stages

    PubMed Central

    Das, Sanchaita; Hehnly, Heidi; Doxsey, Stephen

    2014-01-01

    A recent study revealed new roles for the Rab11 GTPase during mitosis. Rab11 is involved in recycling endosome localization to mitotic spindle poles via dynein-mediated transport. This process is in contrast to Golgi membranes, which disperse in mitosis and do not appear to directly contribute to mitotic functions. Rab11-depletion prevents recycling endosome organization at spindle poles, delays mitotic progression, and disrupts spindle pole protein recruitment, astral microtubule organization, and mitotic spindle orientation. However, Rab11 is not the only endocytic and/or trafficking protein that regulates mitotic progression. Clathrin and two small GTPases (Rab6A’, Rab5) play key roles in spindle organization and function. In this commentary, we discuss the roles of all these canonical endocytic and membrane trafficking proteins during mitosis and speculate on possible cross-communication between them and their molecular pathways that ensure faithful progression through mitosis. PMID:24921241

  15. Identification of a mitotic death signature in cancer cell lines.

    PubMed

    Sakurikar, Nandini; Eichhorn, Joshua M; Alford, Sarah E; Chambers, Timothy C

    2014-02-28

    This study examined the molecular mechanism of action of anti-mitotic drugs. The hypothesis was tested that death in mitosis occurs through sustained mitotic arrest with robust Cdk1 signaling causing complete phosphorylation of Mcl-1 and Bcl-xL, and conversely, that mitotic slippage is associated with incomplete phosphorylation of Mcl-1/Bcl-xL. The results, obtained from studying six different cancer cell lines, strongly support the hypothesis and identify for the first time a unique molecular signature for mitotic death. The findings represent an important advance in understanding anti-mitotic drug action and provide insight into cancer cell susceptibility to such drugs which has important clinical implications. PMID:24099917

  16. MELK is an oncogenic kinase essential for mitotic progression in basal-like breast cancer cells

    PubMed Central

    Wang, Yubao; Lee, Young-Mi; Baitsch, Lukas; Huang, Alan; Xiang, Yi; Tong, Haoxuan; Lako, Ana; Von, Thanh; Choi, Christine; Lim, Elgene; Min, Junxia; Li, Li; Stegmeier, Frank; Schlegel, Robert; Eck, Michael J; Gray, Nathanael S; Mitchison, Timothy J; Zhao, Jean J

    2014-01-01

    Despite marked advances in breast cancer therapy, basal-like breast cancer (BBC), an aggressive subtype of breast cancer usually lacking estrogen and progesterone receptors, remains difficult to treat. In this study, we report the identification of MELK as a novel oncogenic kinase from an in vivo tumorigenesis screen using a kinome-wide open reading frames (ORFs) library. Analysis of clinical data reveals a high level of MELK overexpression in BBC, a feature that is largely dependent on FoxM1, a master mitotic transcription factor that is also found to be highly overexpressed in BBC. Ablation of MELK selectively impairs proliferation of basal-like, but not luminal breast cancer cells both in vitro and in vivo. Mechanistically, depletion of MELK in BBC cells induces caspase-dependent cell death, preceded by defective mitosis. Finally, we find that Melk is not required for mouse development and physiology. Together, these data indicate that MELK is a normally non-essential kinase, but is critical for BBC and thus represents a promising selective therapeutic target for the most aggressive subtype of breast cancer. DOI: http://dx.doi.org/10.7554/eLife.01763.001 PMID:24844244

  17. MELK is an oncogenic kinase essential for mitotic progression in basal-like breast cancer cells.

    PubMed

    Wang, Yubao; Lee, Young-Mi; Baitsch, Lukas; Huang, Alan; Xiang, Yi; Tong, Haoxuan; Lako, Ana; Von, Thanh; Choi, Christine; Lim, Elgene; Min, Junxia; Li, Li; Stegmeier, Frank; Schlegel, Robert; Eck, Michael J; Gray, Nathanael S; Mitchison, Timothy J; Zhao, Jean J

    2014-01-01

    Despite marked advances in breast cancer therapy, basal-like breast cancer (BBC), an aggressive subtype of breast cancer usually lacking estrogen and progesterone receptors, remains difficult to treat. In this study, we report the identification of MELK as a novel oncogenic kinase from an in vivo tumorigenesis screen using a kinome-wide open reading frames (ORFs) library. Analysis of clinical data reveals a high level of MELK overexpression in BBC, a feature that is largely dependent on FoxM1, a master mitotic transcription factor that is also found to be highly overexpressed in BBC. Ablation of MELK selectively impairs proliferation of basal-like, but not luminal breast cancer cells both in vitro and in vivo. Mechanistically, depletion of MELK in BBC cells induces caspase-dependent cell death, preceded by defective mitosis. Finally, we find that Melk is not required for mouse development and physiology. Together, these data indicate that MELK is a normally non-essential kinase, but is critical for BBC and thus represents a promising selective therapeutic target for the most aggressive subtype of breast cancer.DOI: http://dx.doi.org/10.7554/eLife.01763.001. PMID:24844244

  18. Mitotic catastrophe occurs in the absence of apoptosis in p53-null cells with a defective G1 checkpoint.

    PubMed

    Fragkos, Michalis; Beard, Peter

    2011-01-01

    Cell death occurring during mitosis, or mitotic catastrophe, often takes place in conjunction with apoptosis, but the conditions in which mitotic catastrophe may exhibit features of programmed cell death are still unclear. In the work presented here, we studied mitotic cell death by making use of a UV-inactivated parvovirus (adeno-associated virus; AAV) that has been shown to induce a DNA damage response and subsequent death of p53-defective cells in mitosis, without affecting the integrity of the host genome. Osteosarcoma cells (U2OSp53DD) that are deficient in p53 and lack the G1 cell cycle checkpoint respond to AAV infection through a transient G2 arrest. We found that the infected U2OSp53DD cells died through mitotic catastrophe with no signs of chromosome condensation or DNA fragmentation. Moreover, cell death was independent of caspases, apoptosis-inducing factor (AIF), autophagy and necroptosis. These findings were confirmed by time-lapse microscopy of cellular morphology following AAV infection. The assays used readily revealed apoptosis in other cell types when it was indeed occurring. Taken together the results indicate that in the absence of the G1 checkpoint, mitotic catastrophe occurs in these p53-null cells predominantly as a result of mechanical disruption induced by centrosome overduplication, and not as a consequence of a suicide signal. PMID:21853057

  19. Identification of consensus motifs associated with mitotic recombination and clinical characteristics in patients with paternal uniparental isodisomy of chromosome 11.

    PubMed

    Ohtsuka, Yasufumi; Higashimoto, Ken; Oka, Takehiko; Yatsuki, Hitomi; Jozaki, Kosuke; Maeda, Toshiyuki; Kawahara, Kozo; Hamasaki, Yuhei; Matsuo, Muneaki; Nishioka, Kenichi; Joh, Keiichiro; Mukai, Tsunehiro; Soejima, Hidenobu

    2016-04-01

    Uniparental disomy (UPD) is defined as the inheritance of both homologs of a given genomic region from only one parent. The majority of UPD includes an entire chromosome. However, the extent of UPD is sometimes limited to a subchromosomal region (segmental UPD). Mosaic paternal UPD (pUPD) of chromosome 11 is found in approximately 20% of patients with Beckwith-Wiedemann syndrome (BWS) and almost all pUPDs are segmental isodisomic pUPDs resulting from mitotic recombination at an early embryonic stage. A mechanism initiating a DNA double strand break (DSB) within 11p has been predicted to lead to segmental pUPD. However, no consensus motif has yet been found. Here, we analyzed 32 BWS patients with pUPD by SNP array and searched for consensus motifs. We identified four consensus motifs frequently appearing within breakpoint regions of segmental pUPD. These motifs were found in another nine BWS patients with pUPD. In addition, the seven motifs found in meiotic recombination hot spots could not be found within pUPD breakpoint regions. Histone H3 lysine 4 trimethylation, a marker of DSB initiation, could not be found either. These findings suggest that the mechanism(s) of mitotic recombination leading to segmental pUPD are different from that of meiotic recombination. Furthermore, we found seven patients with paternal uniparental diploidy (PUD) mosaicism. Comparison of clinical features between segmental pUPDs and PUDs showed that developmental disability and cardiac abnormalities were additional characteristic features of PUD mosaicism, along with high risk of tumor development. We also found that macroglossia was characteristic of segmental pUPD mosaicism. PMID:26908620

  20. Using high spectral resolution spectrophotometry to study broad mineral absorption features on Mars

    NASA Technical Reports Server (NTRS)

    Blaney, D. L.; Crisp, D.

    1993-01-01

    Traditionally telescopic measurements of mineralogic absorption features have been made using relatively low to moderate (R=30-300) spectral resolution. Mineralogic absorption features tend to be broad so high resolution spectroscopy (R greater than 10,000) does not provide significant additional compositional information. Low to moderate resolution spectroscopy allows an observer to obtain data over a wide wavelength range (hundreds to thousands of wavenumbers) compared to the several wavenumber intervals that are collected using high resolution spectrometers. However, spectrophotometry at high resolution has major advantages over lower resolution spectroscopy in situations that are applicable to studies of the Martian surface, i.e., at wavelengths where relatively weak surface absorption features and atmospheric gas absorption features both occur.

  1. Mitotic checkpoint control and chromatin remodeling.

    PubMed

    Yao, Yixin; Dai, Wei

    2012-01-01

    In order to maintain chromosomal stability during cell division, eukaryotic cells have evolved a number of surveillance mechanisms termed checkpoints. These checkpoints monitor the completion of essential molecular and cellular processes of one stage before entering another. The spindle checkpoint watches the bi-orientation attachment of spindle microtubules to all condensed chromosomes before initiation of nuclear division during mitosis. Histones are subject to a number of post-translational modifications during the cell cycle, which may in turn modify or facilitate cell cycle progression. Recent studies suggest that mitotic proteins including Bub1 and Sgo1 that are involved in the spindle checkpoint also play a major role in the regulation of histone modifications and chromatin remodeling. This mini-review summarizes emerging information about the new role of spindle checkpoint proteins in chromatin remodeling. PMID:22201785

  2. Centromeric DNA cloned from functional kinetochore fragments in mitotic cells with unreplicated genomes.

    PubMed

    Ouspenski, I I; Brinkley, B R

    1993-06-01

    Treatment of cells arrested in the cell cycle at the G1/S-phase boundary with 5 mM caffeine induces premature mitosis, resulting in chromosomal fragmentation and detachment of centromere-kinetochore fragments, which are subsequently attached to the mitotic spindle and segregated in anaphase. Taking advantage of this in vivo separation of the centromere, we have developed a procedure for isolation of a centromere-enriched fraction of mitotic chromatin. Using this method, we have isolated and cloned DNA from the centromere-enriched material of Chinese hamster cells. One of the clones thus obtained was characterized in detail. It contains 6 kb of centromere-associated sequence that exhibits no recognizable homology with other mammalian centromeric sequences and is devoid of any extensive repetitive structure. This sequence is present in a single copy on chromosome 1 and is species-specific. Distinctive features of the clone include the presence of several A+T-rich regions and clusters of multiple topoisomerase II consensus cleavage sites and other sequence motifs characteristic of nuclear matrix-associated regions. We hypothesize that these features might be related to the more compact packaging of centromeric chromatin in interphase nuclei and mitotic chromosomes. PMID:8408270

  3. Phosphohistone H3 expression correlates with manual mitotic counts and aids in identification of "hot spots" in fibroepithelial tumors of the breast.

    PubMed

    Ginter, Paula S; Shin, Sandra J; Liu, Yifang; Chen, Zhengming; D'Alfonso, Timothy M

    2016-03-01

    Classification of mammary fibroepithelial tumors (FETs) relies on assessment of mitotic activity, among other histopathologic parameters. Routine hematoxylin and eosin (H&E) mitotic counts can be subjective and time consuming. Difficulty may arise in identifying "true" mitoses for a variety of reasons. Phosphorylation of histone H3 protein (PHH3) is correlated with mitotic chromatin condensation. The utility of PHH3 immunohistochemical staining to identify mitoses has been demonstrated in multiple organ systems. In this study, we examined the utility of PHH3 in assessing mitotic activity in FETs and compared PHH3- with H&E-determined mitotic counts. PHH3-stained mitoses were readily identifiable at 10 magnification and allowed for rapid identification of mitotic "hot spots." Median mitotic counts/10 high-power fields for fibroadenoma, benign phyllodes tumor, borderline phyllodes tumor (BlnPT), and malignant phyllodes tumor (MPT) were 0, 0.5, 4.25, and 9, respectively on H&E, and 0, 0.75, 4.5, and 8, respectively for PHH3. Among all FETs, there was a strong positive correlation between H&E- and PHH3-determined mitotic counts (r=0.91, P<.001). Using PHH3, 2 cases would be reclassified, both from BlnPT to MPT. PHH3-determined counts correlated with H&E-determined counts in FETs. Using PHH3, a small number of cases were reclassified from BlnPT to MPT, for which treatment is similar. Although H&E-determined counts remain the criterion standard for assessing mitotic activity in FETs, PHH3 may be a useful adjunctive tool in some cases and is helpful in identifying mitotic hot spots. PMID:26826415

  4. The flavonoid eupatorin inactivates the mitotic checkpoint leading to polyploidy and apoptosis

    SciTech Connect

    Salmela, Anna-Leena; Turku Graduate School of Biomedical Sciences, Turku; Turku Centre for Biotechnology, P.O. Box 123, University of Turku ; Pouwels, Jeroen; Kukkonen-Macchi, Anu; Waris, Sinikka; Toivonen, Pauliina; Jaakkola, Kimmo; Maeki-Jouppila, Jenni; Turku Centre for Biotechnology, P.O. Box 123, University of Turku; Drug Discovery Graduate School, University of Turku ; Kallio, Lila; Kallio, Marko J.; Turku Centre for Biotechnology, P.O. Box 123, University of Turku; Centre of Excellence for Translational Genome-Scale Biology, P.O. Box 106, Academy of Finland

    2012-03-10

    The spindle assembly checkpoint (SAC) is a conserved mechanism that ensures the fidelity of chromosome distribution in mitosis by preventing anaphase onset until the correct bipolar microtubule-kinetochore attachments are formed. Errors in SAC function may contribute to tumorigenesis by inducing numerical chromosome anomalies (aneuploidy). On the other hand, total disruption of SAC can lead to massive genomic imbalance followed by cell death, a phenomena that has therapeutic potency. We performed a cell-based high-throughput screen with a compound library of 2000 bioactives for novel SAC inhibitors and discovered a plant-derived phenolic compound eupatorin (3 Prime ,5-dihydroxy-4 Prime ,6,7-trimethoxyflavone) as an anti-mitotic flavonoid. The premature override of the microtubule drug-imposed mitotic arrest by eupatorin is dependent on microtubule-kinetochore attachments but not interkinetochore tension. Aurora B kinase activity, which is essential for maintenance of normal SAC signaling, is diminished by eupatorin in cells and in vitro providing a mechanistic explanation for the observed forced mitotic exit. Eupatorin likely has additional targets since eupatorin treatment of pre-mitotic cells causes spindle anomalies triggering a transient M phase delay followed by impaired cytokinesis and polyploidy. Finally, eupatorin potently induces apoptosis in multiple cancer cell lines and suppresses cancer cell proliferation in organotypic 3D cell culture model.

  5. DEPDC1 is a novel cell cycle related gene that regulates mitotic progression

    PubMed Central

    Mi, Yan; Zhang, Chundong; Bu, Youquan; Zhang, Ying; He, Longxia; Li, Hongxia; Zhu, Huifang; Li, Yi; Lei, Yunlong; Zhu, Jiang

    2015-01-01

    DEPDC1 is a recently identified novel tumor-related gene that is upregulated in several types of cancer and contributes to tumorigenesis. In this study, we have investigated the expression pattern and functional implications of DEPDC1 during cell cycle progression. Expression studies using synchronized cells demonstrated that DEPDC1 is highly expressed in the mitotic phase of the cell cycle. Immunofluorescence assays showed that DEPDC1 is predominantly localized in the nucleus during interphase and is redistributed into the whole cell upon nuclear membrane breakdown in metaphase. Subsequently, siRNA-mediated knockdown of DEPDC1 caused a significant mitotic arrest. Moreover, knockdown of DEPDC1 resulted in remarkable mitotic defects such as abnormal multiple nuclei and multipolar spindle structures accompanied by the upregulation of the A20 gene as well as several cell cycle-related genes such as CCNB1 and CCNB2. Taken together, our current observations strongly suggest that this novel cancerous gene, DEPDC1, plays a pivotal role in the regulation of proper mitotic progression. [BMB Reports 2015; 48(7): 413-418] PMID:25902835

  6. The putative oncogene CEP72 inhibits the mitotic function of BRCA1 and induces chromosomal instability.

    PubMed

    Lüddecke, S; Ertych, N; Stenzinger, A; Weichert, W; Beissbarth, T; Dyczkowski, J; Gaedcke, J; Valerius, O; Braus, G H; Kschischo, M; Bastians, H

    2016-05-01

    BRCA1 is a tumor-suppressor gene associated with, but not restricted to, breast and ovarian cancer and implicated in various biological functions. During mitosis, BRCA1 and its positive regulator Chk2 are localized at centrosomes and are required for the regulation of microtubule plus end assembly, thereby ensuring faithful mitosis and numerical chromosome stability. However, the function of BRCA1 during mitosis has not been defined mechanistically. To gain insights into the mitotic role of BRCA1 in regulating microtubule assembly, we systematically identified proteins interacting with BRCA1 during mitosis and found the centrosomal protein Cep72 as a novel BRCA1-interacting protein. CEP72 is frequently upregulated in colorectal cancer tissues and overexpression of CEP72 mirrors the consequences of BRCA1 loss during mitosis. In detail, the overexpression of CEP72 causes an increase in microtubule plus end assembly, abnormal mitotic spindle formation and the induction of chromosomal instability. Moreover, we show that high levels of Cep72 counteract Chk2 as a positive regulator of BRCA1 to ensure proper mitotic microtubule assembly. Thus, CEP72 represents a putative oncogene in colorectal cancer that might negatively regulate the mitotic function of BRCA1 to ensure chromosomal stability. PMID:26300001

  7. Identification of a novel mitotic phosphorylation motif associated with protein localization to the mitotic apparatus

    SciTech Connect

    Yang, Feng; Camp, David G.; Gritsenko, Marina A.; Luo, Quanzhou; Kelly, Ryan T.; Clauss, Therese RW; Brinkley, William R.; Smith, Richard D.; Stenoien, David L.

    2007-11-16

    The chromosomal passenger complex (CPC) is a critical regulator of chromosome, cytoskeleton and membrane dynamics during mitosis. Here, we identified phosphopeptides and phosphoprotein complexes recognized by a phosphorylation specific antibody that labels the CPC using liquid chromatography coupled to mass spectrometry. A mitotic phosphorylation motif (PX{G/T/S}{L/M}[pS]P or WGL[pS]P) was identified in 11 proteins including Fzr/Cdh1 and RIC-8, two proteins with potential links to the CPC. Phosphoprotein complexes contained known CPC components INCENP, Aurora-B and TD-60, as well as SMAD2, 14-3-3 proteins, PP2A, and Cdk1, a likely kinase for this motif. Protein sequence analysis identified phosphorylation motifs in additional proteins including SMAD2, Plk3 and INCENP. Mitotic SMAD2 and Plk3 phosphorylation was confirmed using phosphorylation specific antibodies, and in the case of Plk3, phosphorylation correlates with its localization to the mitotic apparatus. A mutagenesis approach was used to show INCENP phosphorylation is required for midbody localization. These results provide evidence for a shared phosphorylation event that regulates localization of critical proteins during mitosis.

  8. Genetic variation in mitotic regulatory pathway genes is associated with breast tumor grade

    PubMed Central

    Purrington, Kristen S.; Slettedahl, Seth; Bolla, Manjeet K.; Michailidou, Kyriaki; Czene, Kamila; Nevanlinna, Heli; Bojesen, Stig E.; Andrulis, Irene L.; Cox, Angela; Hall, Per; Carpenter, Jane; Yannoukakos, Drakoulis; Haiman, Christopher A.; Fasching, Peter A.; Mannermaa, Arto; Winqvist, Robert; Brenner, Hermann; Lindblom, Annika; Chenevix-Trench, Georgia; Benitez, Javier; Swerdlow, Anthony; Kristensen, Vessela; Guénel, Pascal; Meindl, Alfons; Darabi, Hatef; Eriksson, Mikael; Fagerholm, Rainer; Aittomäki, Kristiina; Blomqvist, Carl; Nordestgaard, Børge G.; Nielsen, Sune F.; Flyger, Henrik; Wang, Xianshu; Olswold, Curtis; Olson, Janet E.; Mulligan, Anna Marie; Knight, Julia A.; Tchatchou, Sandrine; Reed, Malcolm W.R.; Cross, Simon S.; Liu, Jianjun; Li, Jingmei; Humphreys, Keith; Clarke, Christine; Scott, Rodney; Fostira, Florentia; Fountzilas, George; Konstantopoulou, Irene; Henderson, Brian E.; Schumacher, Fredrick; Le Marchand, Loic; Ekici, Arif B.; Hartmann, Arndt; Beckmann, Matthias W.; Hartikainen, Jaana M.; Kosma, Veli-Matti; Kataja, Vesa; Jukkola-Vuorinen, Arja; Pylkäs, Katri; Kauppila, Saila; Dieffenbach, Aida Karina; Stegmaier, Christa; Arndt, Volker; Margolin, Sara; Balleine, Rosemary; Arias Perez, Jose Ignacio; Pilar Zamora, M.; Menéndez, Primitiva; Ashworth, Alan; Jones, Michael; Orr, Nick; Arveux, Patrick; Kerbrat, Pierre; Truong, Thérèse; Bugert, Peter; Toland, Amanda E.; Ambrosone, Christine B.; Labrèche, France; Goldberg, Mark S.; Dumont, Martine; Ziogas, Argyrios; Lee, Eunjung; Dite, Gillian S.; Apicella, Carmel; Southey, Melissa C.; Long, Jirong; Shrubsole, Martha; Deming-Halverson, Sandra; Ficarazzi, Filomena; Barile, Monica; Peterlongo, Paolo; Durda, Katarzyna; Jaworska-Bieniek, Katarzyna; Tollenaar, Robert A.E.M.; Seynaeve, Caroline; Brüning, Thomas; Ko, Yon-Dschun; Van Deurzen, Carolien H.M.; Martens, John W.M.; Kriege, Mieke; Figueroa, Jonine D.; Chanock, Stephen J.; Lissowska, Jolanta; Tomlinson, Ian; Kerin, Michael J.; Miller, Nicola; Schneeweiss, Andreas; Tapper, William J.; Gerty, Susan M.; Durcan, Lorraine; Mclean, Catriona; Milne, Roger L.; Baglietto, Laura; dos Santos Silva, Isabel; Fletcher, Olivia; Johnson, Nichola; Van'T Veer, Laura J.; Cornelissen, Sten; Försti, Asta; Torres, Diana; Rüdiger, Thomas; Rudolph, Anja; Flesch-Janys, Dieter; Nickels, Stefan; Weltens, Caroline; Floris, Giuseppe; Moisse, Matthieu; Dennis, Joe; Wang, Qin; Dunning, Alison M.; Shah, Mitul; Brown, Judith; Simard, Jacques; Anton-Culver, Hoda; Neuhausen, Susan L.; Hopper, John L.; Bogdanova, Natalia; Dörk, Thilo; Zheng, Wei; Radice, Paolo; Jakubowska, Anna; Lubinski, Jan; Devillee, Peter; Brauch, Hiltrud; Hooning, Maartje; García-Closas, Montserrat; Sawyer, Elinor; Burwinkel, Barbara; Marmee, Frederick; Eccles, Diana M.; Giles, Graham G.; Peto, Julian; Schmidt, Marjanka; Broeks, Annegien; Hamann, Ute; Chang-Claude, Jenny; Lambrechts, Diether; Pharoah, Paul D.P.; Easton, Douglas; Pankratz, V. Shane; Slager, Susan; Vachon, Celine M.; Couch, Fergus J.

    2014-01-01

    Mitotic index is an important component of histologic grade and has an etiologic role in breast tumorigenesis. Several small candidate gene studies have reported associations between variation in mitotic genes and breast cancer risk. We measured associations between 2156 single nucleotide polymorphisms (SNPs) from 194 mitotic genes and breast cancer risk, overall and by histologic grade, in the Breast Cancer Association Consortium (BCAC) iCOGS study (n = 39 067 cases; n = 42 106 controls). SNPs in TACC2 [rs17550038: odds ratio (OR) = 1.24, 95% confidence interval (CI) 1.16–1.33, P = 4.2 × 10−10) and EIF3H (rs799890: OR = 1.07, 95% CI 1.04–1.11, P = 8.7 × 10−6) were significantly associated with risk of low-grade breast cancer. The TACC2 signal was retained (rs17550038: OR = 1.15, 95% CI 1.07–1.23, P = 7.9 × 10−5) after adjustment for breast cancer risk SNPs in the nearby FGFR2 gene, suggesting that TACC2 is a novel, independent genome-wide significant genetic risk locus for low-grade breast cancer. While no SNPs were individually associated with high-grade disease, a pathway-level gene set analysis showed that variation across the 194 mitotic genes was associated with high-grade breast cancer risk (P = 2.1 × 10−3). These observations will provide insight into the contribution of mitotic defects to histological grade and the etiology of breast cancer. PMID:24927736

  9. Dataset from the global phosphoproteomic mapping of early mitotic exit in human cells.

    PubMed

    Rogers, Samuel; McCloy, Rachael A; Parker, Benjamin L; Chaudhuri, Rima; Gayevskiy, Velimir; Hoffman, Nolan J; Watkins, D Neil; Daly, Roger J; James, David E; Burgess, Andrew

    2015-12-01

    The presence or absence of a phosphorylation on a substrate at any particular point in time is a functional readout of the balance in activity between the regulatory kinase and the counteracting phosphatase. Understanding how stable or short-lived a phosphorylation site is required for fully appreciating the biological consequences of the phosphorylation. Our current understanding of kinases and their substrates is well established; however, the role phosphatases play is less understood. Therefore, we utilized a phosphatase dependent model of mitotic exit to identify potential substrates that are preferentially dephosphorylated. Using this method, we identified >16,000 phosphosites on >3300 unique proteins, and quantified the temporal phosphorylation changes that occur during early mitotic exit (McCloy et al., 2015 [1]). Furthermore, we annotated the majority of these phosphorylation sites with a high confidence upstream kinase using published, motif and prediction based methods. The results from this study have been deposited into the ProteomeXchange repository with identifier PXD001559. Here we provide additional analysis of this dataset; for each of the major mitotic kinases we identified motifs that correlated strongly with phosphorylation status. These motifs could be used to predict the stability of phosphorylated residues in proteins of interest, and help infer potential functional roles for uncharacterized phosphorylations. In addition, we provide validation at the single cell level that serine residues phosphorylated by Cdk are stable during phosphatase dependent mitotic exit. In summary, this unique dataset contains information on the temporal mitotic stability of thousands of phosphorylation sites regulated by dozens of kinases, and information on the potential preference that phosphatases have at both the protein and individual phosphosite level. The compellation of this data provides an invaluable resource for the wider research community. PMID:26425664

  10. The centrosome and mitotic spindle apparatus in cancer and senescence.

    PubMed

    Schmidt, Stephen; Essmann, Frank; Cirstea, Ion C; Kuck, Fabian; Thakur, Harish C; Singh, Madhurendra; Kletke, Anja; Jänicke, Reiner U; Wiek, Constanze; Hanenberg, Helmut; Ahmadian, M Reza; Schulze-Osthoff, Klaus; Nürnberg, Bernd; Piekorz, Roland P

    2010-11-15

    Altered cell division is associated with overproliferation and tumorigenesis, however, mitotic aberrations can also trigger antiproliferative responses leading to postmitotic cell cycle exit. Here, we focus on the role of the centrosome and in particular of centrosomal TACC (transforming acidic coiled coil) proteins in tumorigenesis and cellular senescence. We have complied recent evidence that inhibition or depletion of various mitotic proteins which take over key in centrosome and kinetochore integrity and mitotic checkpoint function in sufficient to activate a p53-p21(WAF) driven premature senescence phenotype. These findings have direct implications for proliferative tissue homeostasis as well as for cellular and organismal aging. PMID:21088502

  11. Plk1 Inhibition Causes Post-Mitotic DNA Damage and Senescence in a Range of Human Tumor Cell Lines

    PubMed Central

    Bowman, Doug; Shinde, Vaishali; Lasky, Kerri; Shi, Judy; Vos, Tricia; Stringer, Bradley; Amidon, Ben; D'Amore, Natalie; Hyer, Marc L.

    2014-01-01

    Plk1 is a checkpoint protein whose role spans all of mitosis and includes DNA repair, and is highly conserved in eukaryotes from yeast to man. Consistent with this wide array of functions for Plk1, the cellular consequences of Plk1 disruption are diverse, spanning delays in mitotic entry, mitotic spindle abnormalities, and transient mitotic arrest leading to mitotic slippage and failures in cytokinesis. In this work, we present the in vitro and in vivo consequences of Plk1 inhibition in cancer cells using potent, selective small-molecule Plk1 inhibitors and Plk1 genetic knock-down approaches. We demonstrate for the first time that cellular senescence is the predominant outcome of Plk1 inhibition in some cancer cell lines, whereas in other cancer cell lines the dominant outcome appears to be apoptosis, as has been reported in the literature. We also demonstrate strong induction of DNA double-strand breaks in all six lines examined (as assayed by γH2AX), which occurs either during mitotic arrest or mitotic-exit, and may be linked to the downstream induction of senescence. Taken together, our findings expand the view of Plk1 inhibition, demonstrating the occurrence of a non-apoptotic outcome in some settings. Our findings are also consistent with the possibility that mitotic arrest observed as a result of Plk1 inhibition is at least partially due to the presence of unrepaired double-strand breaks in mitosis. These novel findings may lead to alternative strategies for the development of novel therapeutic agents targeting Plk1, in the selection of biomarkers, patient populations, combination partners and dosing regimens. PMID:25365521

  12. Plk1 inhibition causes post-mitotic DNA damage and senescence in a range of human tumor cell lines.

    PubMed

    Driscoll, Denise L; Chakravarty, Arijit; Bowman, Doug; Shinde, Vaishali; Lasky, Kerri; Shi, Judy; Vos, Tricia; Stringer, Bradley; Amidon, Ben; D'Amore, Natalie; Hyer, Marc L

    2014-01-01

    Plk1 is a checkpoint protein whose role spans all of mitosis and includes DNA repair, and is highly conserved in eukaryotes from yeast to man. Consistent with this wide array of functions for Plk1, the cellular consequences of Plk1 disruption are diverse, spanning delays in mitotic entry, mitotic spindle abnormalities, and transient mitotic arrest leading to mitotic slippage and failures in cytokinesis. In this work, we present the in vitro and in vivo consequences of Plk1 inhibition in cancer cells using potent, selective small-molecule Plk1 inhibitors and Plk1 genetic knock-down approaches. We demonstrate for the first time that cellular senescence is the predominant outcome of Plk1 inhibition in some cancer cell lines, whereas in other cancer cell lines the dominant outcome appears to be apoptosis, as has been reported in the literature. We also demonstrate strong induction of DNA double-strand breaks in all six lines examined (as assayed by γH2AX), which occurs either during mitotic arrest or mitotic-exit, and may be linked to the downstream induction of senescence. Taken together, our findings expand the view of Plk1 inhibition, demonstrating the occurrence of a non-apoptotic outcome in some settings. Our findings are also consistent with the possibility that mitotic arrest observed as a result of Plk1 inhibition is at least partially due to the presence of unrepaired double-strand breaks in mitosis. These novel findings may lead to alternative strategies for the development of novel therapeutic agents targeting Plk1, in the selection of biomarkers, patient populations, combination partners and dosing regimens. PMID:25365521

  13. Dissociation of gemcitabine chemosensitization by CHK1 inhibition from cell cycle checkpoint abrogation and aberrant mitotic entry.

    PubMed

    Parsels, Leslie A; Tanska, Daria M; Parsels, Joshua D; Zabludoff, Sonya D; Cuneo, Kyle C; Lawrence, Theodore S; Maybaum, Jonathan; Morgan, Meredith A

    2016-03-01

    In order to determine the relative contribution of checkpoint abrogation and subsequent aberrant mitotic entry to gemcitabine chemosensitization by CHK1 inhibition, we established a model utilizing the CDK inhibitors roscovitine or purvalanol A to re-establish cell cycle arrest and prevent aberrant mitotic entry in pancreatic cancer cells treated with gemcitabine and the CHK inhibitor AZD7762. In this study, we report that the extent of aberrant mitotic entry, as determined by flow cytometry for the mitotic marker phospho-Histone H3 (Ser10), did not reflect the relative sensitivities of pancreatic cancer cell lines to gemcitabine chemosensitization by AZD7762. In addition, re-establishing gemcitabine-induced cell cycle arrest either pharmacologically, with roscovitine or purvalanol A, or genetically, with cyclin B1 siRNA, did not inhibit chemosensitization uniformly across the cell lines. Furthermore, we found that AZD7762 augmented high-intensity γH2AX signaling in gemcitabine-treated cells, suggesting the presence of replication stress when CHK1 is inhibited. Finally, the ability of roscovitine to prevent chemosensitization correlated with its ability to inhibit AZD7762-induced high-intensity γH2AX, but not aberrant pHH3, suggesting that the effects of AZD7762 on DNA replication or repair rather than aberrant mitotic entry determine gemcitabine chemosensitization in pancreatic cancer cells. PMID:26890478

  14. Comparative diagnostic and prognostic performances of the hematoxylin-eosin and phospho-histone H3 mitotic count and Ki-67 index in adrenocortical carcinoma.

    PubMed

    Duregon, Eleonora; Molinaro, Luca; Volante, Marco; Ventura, Laura; Righi, Luisella; Bolla, Stefania; Terzolo, Massimo; Sapino, Anna; Papotti, Mauro G

    2014-09-01

    Mitotic count on hematoxylin and eosin slides is a fundamental morphological criterion in the diagnosis and grading of adrenocortical carcinoma in any scoring system employed. Moreover, it is the unique term strongly associated with patient's prognosis. Phospho-histone H3 is a mitosis-specific antibody, which was already proven to facilitate mitotic count in melanoma and other tumors. Therefore, a study was designed to assess the diagnostic and prognostic role of phospho-histone H3 in 52 adrenocortical carcinomas, comparing manual and computerized count to standard manual hematoxylin- and eosin-based method and Ki-67 index. Manual hematoxylin and eosin and phospho-histone H3 mitotic counts were highly correlated (r=0.9077, P<0.0001), better than computer-assisted phospho-histone H3 evaluations, and had an excellent inter-observer reproducibility at Bland-Altman analysis. Three of 15 cases having <5 mitotic figures per 50 high-power fields by standard count on hematoxylin and eosin gained the mitotic figure point of Weiss Score after a manual count on phospho-histone H3 slides. Traditional mitotic count confirmed to be a strong predictor of overall survival (P=0.0043), better than phospho-histone H3-based evaluation (P=0.051), but not as strong as the Ki-67 index (P<0.0001). The latter further segregated adrenocortical carcinomas into three prognostic groups, stratifying cases by low (<20%), intermediate (20-50%), and high (>50%) Ki-67 values. We conclude that (a) phospho-histone H3 staining is a useful diagnostic complementary tool to standard hematoxylin and eosin mitotic count, enabling optimal mitotic figure evaluation (including atypical mitotic figures) even in adrenocortical carcinomas with a low mitotic index and with a very high reproducibility; (b) Ki-67 proved to be the best prognostic indicator of overall survival, being superior to the mitotic index, irrespective of the method (standard on hematoxylin and eosin or phospho-histone H3-based) used to count mitotic figures. PMID:24434900

  15. Human LATS1 is a mitotic exit network kinase.

    PubMed

    Bothos, John; Tuttle, Robyn L; Ottey, Michelle; Luca, Francis C; Halazonetis, Thanos D

    2005-08-01

    The kinase LATS/WARTS is a tumor suppressor protein conserved in evolution, but its function at the molecular level is not well understood. We report here that human LATS1 interacts with MOB1A, a protein whose homologue in budding yeast associates with kinases involved in mitotic exit. This suggested that LATS1 may be a component of the previously uncharacterized mitotic exit network in higher eukaryotes. Indeed, moderate overexpression of human LATS1 in cells exposed to microtubule poisons facilitated mitotic exit, and this activity required MOB1A. Reciprocally, small interfering RNA-mediated suppression of LATS1 or MOB1A prolonged telophase, but had no effect on the length of the earlier phases of mitosis. A role of LATS1 in mitotic exit may explain its previously described abilities to induce G2 arrest and promote cytokinesis. PMID:16061636

  16. Centrosomes and mitotic spindle poles: a recent liaison?

    PubMed

    Chavali, Pavithra L; Peset, Isabel; Gergely, Fanni

    2015-02-01

    Centrosomes comprise two cylindrical centrioles embedded in the pericentriolar material (PCM). The PCM is an ordered assembly of large scaffolding molecules, providing an interaction platform for proteins involved in signalling, trafficking and most importantly microtubule nucleation and organization. In mitotic cells, centrosomes are located at the spindle poles, sites where spindle microtubules converge. However, certain cell types and organisms lack centrosomes, yet contain focused spindle poles, highlighting that despite their juxtaposition in cells, centrosomes and mitotic spindle poles are distinct physical entities. In the present paper, we discuss the origin of centrosomes and summarize their contribution to mitotic spindle assembly and cell division. We then describe the key molecular players that mediate centrosome attachment to mitotic spindle poles and explore why co-segregation of centrosomes and spindle poles into daughter cells is of potential benefit to organisms. PMID:25619241

  17. The bipolar assembly domain of the mitotic motor kinesin-5

    PubMed Central

    Acar, Seyda; Carlson, David B.; Budamagunta, Madhu S.; Yarov-Yarovoy, Vladimir; Correia, John J.; Niñonuevo, Milady R.; Jia, Weitao; Tao, Li; Leary, Julie A.; Voss, John C.; Evans, James E.; Scholey, Jonathan M.

    2013-01-01

    An outstanding unresolved question is how does the mitotic spindle utilize microtubules and mitotic motors to coordinate accurate chromosome segregation during mitosis? This process depends upon the mitotic motor, kinesin-5, whose unique bipolar architecture, with pairs of motor domains lying at opposite ends of a central rod, allows it to crosslink microtubules within the mitotic spindle and to coordinate their relative sliding during spindle assembly, maintenance and elongation. The structural basis of kinesin-5’s bipolarity is, however, unknown, as protein asymmetry has so far precluded its crystallization. Here we use electron microscopy of single molecules of kinesin-5 and its subfragments, combined with hydrodynamic analysis plus mass spectrometry, circular dichroism and site-directed spin label electron paramagnetic resonance spectroscopy, to show how a staggered antiparallel coiled-coil ‘BASS’ (bipolar assembly) domain directs the assembly of four kinesin-5 polypeptides into bipolar minifilaments. PMID:23299893

  18. The bipolar assembly domain of the mitotic motor kinesin-5.

    PubMed

    Acar, Seyda; Carlson, David B; Budamagunta, Madhu S; Yarov-Yarovoy, Vladimir; Correia, John J; Niñonuevo, Milady R; Jia, Weitao; Tao, Li; Leary, Julie A; Voss, John C; Evans, James E; Scholey, Jonathan M

    2013-01-01

    An outstanding unresolved question is how does the mitotic spindle utilize microtubules and mitotic motors to coordinate accurate chromosome segregation during mitosis? This process depends upon the mitotic motor, kinesin-5, whose unique bipolar architecture, with pairs of motor domains lying at opposite ends of a central rod, allows it to crosslink microtubules within the mitotic spindle and to coordinate their relative sliding during spindle assembly, maintenance and elongation. The structural basis of kinesin-5's bipolarity is, however, unknown, as protein asymmetry has so far precluded its crystallization. Here we use electron microscopy of single molecules of kinesin-5 and its subfragments, combined with hydrodynamic analysis plus mass spectrometry, circular dichroism and site-directed spin label electron paramagnetic resonance spectroscopy, to show how a staggered antiparallel coiled-coil 'BASS' (bipolar assembly) domain directs the assembly of four kinesin-5 polypeptides into bipolar minifilaments. PMID:23299893

  19. Sister chromatid exchanges and mitotic crossing-over

    SciTech Connect

    1993-12-31

    Chapter 12, discusses sister chromatid exchanges (SCE) and mitotic crossing-over. The detection, occurrence and use of SCE in mutagenesis research is covered, as is segregation, chiasmata and gene amplification. 28 refs., 5 figs.

  20. Mitotic Diversity in Homeostatic Human Interfollicular Epidermis

    PubMed Central

    Nöske, Katharina; Stark, Hans-Jürgen; Nevaril, Leonard; Berning, Manuel; Langbein, Lutz; Goyal, Ashish; Diederichs, Sven; Boukamp, Petra

    2016-01-01

    Despite decades of skin research, regulation of proliferation and homeostasis in human epidermis is still insufficiently understood. To address the role of mitoses in tissue regulation, we utilized human long-term skin equivalents and systematically assessed mitoses during early epidermal development and long-term epidermal regeneration. We now demonstrate four different orientations: (1) horizontal, i.e., parallel to the basement membrane (BM) and suggestive of symmetric divisions; (2) oblique with an angle of 45°–70°; or (3) perpendicular, suggestive of asymmetric division. In addition, we demonstrate a fourth substantial fraction of suprabasal mitoses, many of which are committed to differentiation (Keratin K10-positive). As verified also for normal human skin, this spatial mitotic organization is part of the regulatory program of human epidermal tissue homeostasis. As a potential marker for asymmetric division, we investigated for Numb and found that it was evenly spread in almost all undifferentiated keratinocytes, but indeed asymmetrically distributed in some mitoses and particularly frequent under differentiation-repressing low-calcium conditions. Numb deletion (stable knockdown by CRISPR/Cas9), however, did not affect proliferation, neither in a three-day follow up study by life cell imaging nor during a 14-day culture period, suggesting that Numb is not essential for the general control of keratinocyte division. PMID:26828486

  1. Fabrication of a high-resolution roll for gravure printing of 2μm features

    NASA Astrophysics Data System (ADS)

    Grau, Gerd; Kitsomboonloha, Rungrot; Subramanian, Vivek

    2015-08-01

    High-resolution features are key to achieve high performance printed electronics devices such as transistors. Gravure printing is very promising to achieve high resolution in combination with high printing speeds on the order of 1m/s. High-speed gravure has recently been shown to print high resolution features down to linewidths and spacing of 2μm. Whilst this was a tremendous improvement over previous reports, these results had been obtained using silicon printing plates. These silicon printing plates are fabricated using microfabrication techniques which offer several advantages over traditional metal gravure cylinders where the features are defined by techniques such as stylus engraving, laser engraving or etching. This offers much greater precision and design freedom in terms of feature size, surface roughness, cell placement and cell shape. However, rigid silicon printing plates cannot be used in a roll-to-roll printing process that would truly enable low-cost printed electronics. Here we demonstrate for the first time a gravure printing roll that combines the precision of silicon printing plates with the form factor of a metal cylinder. The fabrication process starts with a silicon master whose pattern is replicated by polymer molding. The actual metal printing plate is then built up on the polymer negative of the pattern by a combination of electroless and electroplating. After separation of the polymer and the metal, the metal printing plate can be mounted on a magnetic roll for printing. Printing of highly scaled 2μm features is demonstrated. Different metal surfaces were explored to optimize printing performance and wear during printing.

  2. Physical limits on kinesin-5–mediated chromosome congression in the smallest mitotic spindles

    PubMed Central

    McCoy, Kelsey M.; Tubman, Emily S.; Claas, Allison; Tank, Damien; Clancy, Shelly Applen; O’Toole, Eileen T.; Berman, Judith; Odde, David J.

    2015-01-01

    A characteristic feature of mitotic spindles is the congression of chromosomes near the spindle equator, a process mediated by dynamic kinetochore microtubules. A major challenge is to understand how precise, submicrometer-scale control of kinetochore micro­tubule dynamics is achieved in the smallest mitotic spindles, where the noisiness of microtubule assembly/disassembly will potentially act to overwhelm the spatial information that controls microtubule plus end–tip positioning to mediate congression. To better understand this fundamental limit, we conducted an integrated live fluorescence, electron microscopy, and modeling analysis of the polymorphic fungal pathogen Candida albicans, which contains one of the smallest known mitotic spindles (<1 μm). Previously, ScCin8p (kinesin-5 in Saccharomyces cerevisiae) was shown to mediate chromosome congression by promoting catastrophe of long kinetochore microtubules (kMTs). Using C. albicans yeast and hyphal kinesin-5 (Kip1p) heterozygotes (KIP1/kip1∆), we found that mutant spindles have longer kMTs than wild-type spindles, consistent with a less-organized spindle. By contrast, kinesin-8 heterozygous mutant (KIP3/kip3∆) spindles exhibited the same spindle organization as wild type. Of interest, spindle organization in the yeast and hyphal states was indistinguishable, even though yeast and hyphal cell lengths differ by two- to fivefold, demonstrating that spindle length regulation and chromosome congression are intrinsic to the spindle and largely independent of cell size. Together these results are consistent with a kinesin-5–mediated, length-dependent depolymerase activity that organizes chromosomes at the spindle equator in C. albicans to overcome fundamental noisiness in microtubule self-assembly. More generally, we define a dimensionless number that sets a fundamental physical limit for maintaining congression in small spindles in the face of assembly noise and find that C. albicans operates very close to this limit, which may explain why it has the smallest known mitotic spindle that still manifests the classic congression architecture. PMID:26354423

  3. Application of high-dimensional feature selection: evaluation for genomic prediction in man.

    PubMed

    Bermingham, M L; Pong-Wong, R; Spiliopoulou, A; Hayward, C; Rudan, I; Campbell, H; Wright, A F; Wilson, J F; Agakov, F; Navarro, P; Haley, C S

    2015-01-01

    In this study, we investigated the effect of five feature selection approaches on the performance of a mixed model (G-BLUP) and a Bayesian (Bayes C) prediction method. We predicted height, high density lipoprotein cholesterol (HDL) and body mass index (BMI) within 2,186 Croatian and into 810 UK individuals using genome-wide SNP data. Using all SNP information Bayes C and G-BLUP had similar predictive performance across all traits within the Croatian data, and for the highly polygenic traits height and BMI when predicting into the UK data. Bayes C outperformed G-BLUP in the prediction of HDL, which is influenced by loci of moderate size, in the UK data. Supervised feature selection of a SNP subset in the G-BLUP framework provided a flexible, generalisable and computationally efficient alternative to Bayes C; but careful evaluation of predictive performance is required when supervised feature selection has been used. PMID:25988841

  4. Application of high-dimensional feature selection: evaluation for genomic prediction in man

    PubMed Central

    Bermingham, M. L.; Pong-Wong, R.; Spiliopoulou, A.; Hayward, C.; Rudan, I.; Campbell, H.; Wright, A. F.; Wilson, J. F.; Agakov, F.; Navarro, P.; Haley, C. S.

    2015-01-01

    In this study, we investigated the effect of five feature selection approaches on the performance of a mixed model (G-BLUP) and a Bayesian (Bayes C) prediction method. We predicted height, high density lipoprotein cholesterol (HDL) and body mass index (BMI) within 2,186 Croatian and into 810 UK individuals using genome-wide SNP data. Using all SNP information Bayes C and G-BLUP had similar predictive performance across all traits within the Croatian data, and for the highly polygenic traits height and BMI when predicting into the UK data. Bayes C outperformed G-BLUP in the prediction of HDL, which is influenced by loci of moderate size, in the UK data. Supervised feature selection of a SNP subset in the G-BLUP framework provided a flexible, generalisable and computationally efficient alternative to Bayes C; but careful evaluation of predictive performance is required when supervised feature selection has been used. PMID:25988841

  5. Fully functional global genome repair of (6-4) photoproducts and compromised transcription-coupled repair of cyclobutane pyrimidine dimers in condensed mitotic chromatin

    SciTech Connect

    Komura, Jun-ichiro; Ikehata, Hironobu; Mori, Toshio; Ono, Tetsuya

    2012-03-10

    During mitosis, chromatin is highly condensed, and activities such as transcription and semiconservative replication do not occur. Consequently, the condensed condition of mitotic chromatin is assumed to inhibit DNA metabolism by impeding the access of DNA-transacting proteins. However, about 40 years ago, several researchers observed unscheduled DNA synthesis in UV-irradiated mitotic chromosomes, suggesting the presence of excision repair. We re-examined this subject by directly measuring the removal of UV-induced DNA lesions by an ELISA and by a Southern-based technique in HeLa cells arrested at mitosis. We observed that the removal of (6-4) photoproducts from the overall genome in mitotic cells was as efficient as in interphase cells. This suggests that global genome repair of (6-4) photoproducts is fully functional during mitosis, and that the DNA in mitotic chromatin is accessible to proteins involved in this mode of DNA repair. Nevertheless, not all modes of DNA repair seem fully functional during mitosis. We also observed that the removal of cyclobutane pyrimidine dimers from the dihydrofolate reductase and c-MYC genes in mitotic cells was very slow. This suggests that transcription-coupled repair of cyclobutane pyrimidine dimers is compromised or non-functional during mitosis, which is probably the consequence of mitotic transcriptional repression. -- Highlights: Black-Right-Pointing-Pointer Global genome repair of (6-4) photoproducts is fully active in mitotic cells. Black-Right-Pointing-Pointer DNA in condensed mitotic chromatin does not seem inaccessible or inert. Black-Right-Pointing-Pointer Mitotic transcriptional repression may impair transcription-coupled repair.

  6. Micromechanical-biochemical studies of mitotic chromosome elasticity and structure

    NASA Astrophysics Data System (ADS)

    Poirier, Michael Guy

    The structure of mitotic chromosomes was studied by combining micromechanical force measurements with microfluidic biochemical exposures. Our method is to use glass micropipettes attached to either end of a single chromosome to do mechanical experiments in the extracellular buffer. A third pipette can be used to locally 'spray' reactants so as to carry out dynamical mechanical-chemical experiments. The following elastic properties of mitotic chromosomes are found: Young's modulus, Y = 300 Pa; Poisson ratio, sigma = 0.1; Bending rigidity, B = 1 x 10 -22 J·m; Internal viscosity, eta' = 100 kg/m·sec; Volume fraction, ϕ = 0.7; Extensions of less than 3 times the relaxed length are linear and reversible; Extensions beyond 30 fold exhibit a force plateau at 15 nN and convert the chromosome to a disperse ghost-like state with little change in chromatin structure; Mitotic chromosomes are relatively isotropic; dsDNA cuts of at least every 3 kb cause the a mitotic chromosomes to fall apart; dsDNA cuts less frequently than every 50 kb do not affect mitotic chromosome structure. These results lead to the conclusion that mitotic chromosomes are a network crosslinked every 50 kb between which chromatin is fold by chromatin folding proteins, which are likely to be condensins.

  7. Timeless links replication termination to mitotic kinase activation.

    PubMed

    Dheekollu, Jayaraju; Wiedmer, Andreas; Hayden, James; Speicher, David; Gotter, Anthony L; Yen, Tim; Lieberman, Paul M

    2011-01-01

    The mechanisms that coordinate the termination of DNA replication with progression through mitosis are not completely understood. The human Timeless protein (Tim) associates with S phase replication checkpoint proteins Claspin and Tipin, and plays an important role in maintaining replication fork stability at physical barriers, like centromeres, telomeres and ribosomal DNA repeats, as well as at termination sites. We show here that human Tim can be isolated in a complex with mitotic entry kinases CDK1, Auroras A and B, and Polo-like kinase (Plk1). Plk1 bound Tim directly and colocalized with Tim at a subset of mitotic structures in M phase. Tim depletion caused multiple mitotic defects, including the loss of sister-chromatid cohesion, loss of mitotic spindle architecture, and a failure to exit mitosis. Tim depletion caused a delay in mitotic kinase activity in vivo and in vitro, as well as a reduction in global histone H3 S10 phosphorylation during G2/M phase. Tim was also required for the recruitment of Plk1 to centromeric DNA and formation of catenated DNA structures at human centromere alpha satellite repeats. Taken together, these findings suggest that Tim coordinates mitotic kinase activation with termination of DNA replication. PMID:21573113

  8. Targeting the Mitotic Catastrophe Signaling Pathway in Cancer

    PubMed Central

    Mc Gee, Margaret M.

    2015-01-01

    Mitotic catastrophe, as defined in 2012 by the International Nomenclature Committee on Cell Death, is a bona fide intrinsic oncosuppressive mechanism that senses mitotic failure and responds by driving a cell to an irreversible antiproliferative fate of death or senescence. Thus, failed mitotic catastrophe can promote the unrestrained growth of defective cells, thereby representing a major gateway to tumour development. Furthermore, the activation of mitotic catastrophe offers significant therapeutic advantage which has been exploited in the action of conventional and targeted anticancer agents. Yet, despite its importance in tumour prevention and treatment, the molecular mechanism of mitotic catastrophe is not well understood. A better understanding of the signals that determine cell fate following failed or defective mitosis will reveal new opportunities to selectively target and enhance the programme for therapeutic benefit and reveal biomarkers to predict patient response. This review is focused on the molecular mechanism of mitotic catastrophe induction and signalling and highlights current strategies to exploit the process in cancer therapy. PMID:26491220

  9. Sharing of mitotic pre-ribosomal particles between daughter cells.

    PubMed

    Sirri, Valentina; Jourdan, Nathalie; Hernandez-Verdun, Danièle; Roussel, Pascal

    2016-04-15

    Ribosome biogenesis is a fundamental multistep process initiated by the synthesis of 90S pre-ribosomal particles in the nucleoli of higher eukaryotes. Even though synthesis of ribosomes stops during mitosis while nucleoli disappear, mitotic pre-ribosomal particles persist as observed in pre-nucleolar bodies (PNBs) during telophase. To further understand the relationship between the nucleolus and the PNBs, the presence and the fate of the mitotic pre-ribosomal particles during cell division were investigated. We demonstrate that the recently synthesized 45S precursor ribosomal RNAs (pre-rRNAs) as well as the 32S and 30S pre-rRNAs are maintained during mitosis and associated with the chromosome periphery together with pre-rRNA processing factors. Maturation of the mitotic pre-ribosomal particles, as assessed by the stability of the mitotic pre-rRNAs, is transiently arrested during mitosis by a cyclin-dependent kinase (CDK)1-cyclin-B-dependent mechanism and can be restored by CDK inhibitor treatments. At the M-G1 transition, the resumption of mitotic pre-rRNA processing in PNBs does not induce the disappearance of PNBs; this only occurs when functional nucleoli reform. Strikingly, during their maturation process, mitotic pre-rRNAs localize in reforming nucleoli. PMID:26929073

  10. High-accuracy real-time pedestrian detection system using 2D and 3D features

    NASA Astrophysics Data System (ADS)

    Chambers, David R.; Flannigan, Clay; Wheeler, Benjamin

    2012-06-01

    We present a real time stereo-vision pedestrian detector implementation with a very high accuracy, the 2D component of which attains 99% recall with less than 10-6 false positives per window on the INRIA persons dataset. We utilize a sequence of classifiers which use different features, beginning with Haar-like features and a Haar-like feature implementation adapted to disparity images, and performing a final verification with Histogram-of-Oriented Gradient (HOG) features. We present a 2D Haar-like feature implementation that utilizes 2x2 kernel filters at multiple scales rather than integral images, and combines a quickly trained preliminary adaBoost classifier with a more accurate SVM classifier. We also show how these Haar-like features may be computed from a partially incomplete stereo disparity image in order to make use of 3-dimensional data. Finally, we discuss how these features, along with the HOG features, are computed rapidly and how the classifiers are combined in such a way as to enable real-time implementation with higher detection rates and lower false positive rates than typical systems. Our overall detector is a practical combination of speed and detection performance, operating on 544x409 image (10,425 windows) at a frame rate of 10-20fps, depending on scene complexity. The detector's overall false positive rate is less than 10-6, corresponding to about one false positive every 10-60s when testing on our non-training data. Additionally, the detector has shown usefulness for detecting other object types, and has been implemented for traffic cones, telephone poles, and vehicles.

  11. Distinct clinicopathological features of NAB2-STAT6 fusion gene variants in solitary fibrous tumor with emphasis on the acquisition of highly malignant potential.

    PubMed

    Akaike, Keisuke; Kurisaki-Arakawa, Aiko; Hara, Kieko; Suehara, Yoshiyuki; Takagi, Tatsuya; Mitani, Keiko; Kaneko, Kazuo; Yao, Takashi; Saito, Tsuyoshi

    2015-03-01

    The impact of NGFI-A binding protein 2 (NAB2)-signal transducer and activator of transcription 6 (STAT6) fusion on the biological behavior and the mechanism of acquisition of malignant phenotype in solitary fibrous tumor (SFT) is not well understood. We examined variations of the NAB2-STAT6 fusion gene in 40 cases of SFT using formalin-fixed, paraffin-embedded tissues and secondary genetic alterations of tumor protein p53 (TP53),, platelet-derived growth factor receptor, β polypeptide (PDGFRB), and telomerase reverse transcriptase (TERT) promoters. These gene variations were compared with the clinicopathological features. The 2-year and 5-year disease-free survival rates (DFSRs) were 91% and 83%, respectively. All 40 samples demonstrated nuclear staining for STAT6, including CD34-negative cases. Moreover, p53-positive staining was associated with a lower DFSR and was significantly associated with higher Ki-67 label index, higher mitotic rate (mitosis, >4/high-power field), and the presence of nuclear atypia/pleomorphism. NAB2-STAT6 fusions were detected in all of the cases; the NAB2 exon 4-STAT6 exon 2, the most common genotype, appeared in 18 cases, which was associated with thoracic tumor location and the less aggressive phenotype. In contrast, tumors with NAB2 exon 6-STAT6 exon 16/18 demonstrated an aggressive phenotype. Mutations in TP53 and PDGFRB were detected in 2 and 3 cases respectively, and these occurred in a mutually exclusive fashion. TERT promoter hot spot mutations were observed in 5 cases, which were associated with shorter DFSR. Two dedifferentiated SFT cases harbored both TP53 and TERT promoter mutations. TP53 mutations, which result in its overexpression, in combination with TERT promoter mutations seem to play an important role in the dedifferentiation process. PMID:25582503

  12. Access, Participation, and Supports: The Defining Features of High-Quality Inclusion

    ERIC Educational Resources Information Center

    Buysse, Virginia

    2011-01-01

    This article describes current knowledge about early childhood inclusion, summarizing research and the DEC/NAEYC joint position statement on inclusion. The article also describes effective or promising educational practices that promote access, participation, and supports--the defining features of high-quality inclusion. Future efforts to improve…

  13. Assessing Motor Skills as a Differentiating Feature between High Functioning Autism and Asperger's Disorder

    ERIC Educational Resources Information Center

    Cid, Maria R.

    2011-01-01

    The purpose of this research was to investigate if motor skills could be used as a differentiating feature between Asperger's Disorder (AD) and High Functioning (HFA) in children under the age of 9 years, 0 months, in order to provide additional information regarding the usefulness and validity of distinguishing these two disorders. There is…

  14. Interaction between TBP and Condensin Drives the Organization and Faithful Segregation of Mitotic Chromosomes.

    PubMed

    Iwasaki, Osamu; Tanizawa, Hideki; Kim, Kyoung-Dong; Yokoyama, Yuhki; Corcoran, Christopher J; Tanaka, Atsunari; Skordalakes, Emmanuel; Showe, Louise C; Noma, Ken-Ichi

    2015-09-01

    Genome/chromosome organization is highly ordered and controls various nuclear events, although the molecular mechanisms underlying the functional organization remain largely unknown. Here, we show that the TATA box-binding protein (TBP) interacts with the Cnd2 kleisin subunit of condensin to mediate interphase and mitotic chromosomal organization in fission yeast. TBP recruits condensin onto RNA polymerase III-transcribed (Pol III) genes and highly transcribed Pol II genes; condensin in turn associates these genes with centromeres. Inhibition of the Cnd2-TBP interaction disrupts condensin localization across the genome and the proper assembly of mitotic chromosomes, leading to severe defects in chromosome segregation and eventually causing cellular lethality. We propose that the Cnd2-TBP interaction coordinates transcription with chromosomal architecture by linking dispersed gene loci with centromeres. This chromosome arrangement can contribute to the efficient transmission of physical force at the kinetochore to chromosomal arms, thereby supporting the fidelity of chromosome segregation. PMID:26257282

  15. Tectonic Features in the Equatorial Lowlands of Mercury Viewed at High Incidence Angles

    NASA Astrophysics Data System (ADS)

    Selvans, M. M.; Watters, T. R.; Solomon, S. C.

    2012-12-01

    The spatial distribution of tectonic features on Mercury, although not fully understood, is related to the stress regime and the mechanical properties of the lithosphere during the time that the features formed and remained active. Lobate scarps and high-relief ridges, compressional features that generally have ~1 km of relief and are hundreds of kilometers long, were identified on Mercury from images acquired during the Mariner 10 and MErcury Surface, Space ENvironment, GEochemistry, and Ranging (MESSENGER) flybys. Images taken from orbit during the primary MESSENGER mission, with full coverage of the surface, confirmed that these scarps and ridges appear to be concentrated in three broad, north-south bands. Images at high incidence angles, collected since April 2012 during the MESSENGER extended mission, provide a more complete picture of the spatial extent and orientations of these features, and of their relationship to neighboring landforms. Digital elevation models, from laser altimetry and stereo imaging, additionally allow for comparisons between tectonic landforms and elevation and for measurements of slope and relief across individual features. Scarps and ridges are found at a wide range of elevations on Mercury. The greatest concentration of such features in an equatorial lowland setting is in an area (40°N-40°S, 220°-270°E) that is within one of the three north-south bands of tectonic features. Within this area, the 48 previously mapped features generally do not display preferred orientations or a consistent relationship to topography. Of these scarps, 47 were identified in flyby images and one in orbital images. Three follow the rim of Beethoven basin (10°-30°S, 225-245°E, ~600 km diameter), likely having formed along earlier zones of weakness in the crust created during formation of the basin. From recent images taken at high incidence angles, which currently have ~75% coverage in this equatorial lowland area, we are able to identify only seven additional tectonic features, all within Beethoven basin. Six of these newly identified features are also subparallel to the basin rim. However, no other scarps in our study area are so clearly connected to a particular topographic or geologic feature. The 22 lobate scarps and high-relief ridges in the northeastern quadrant of our study area have similar base elevations (average of -0.78 km, standard deviation of 0.17 km) and relief. Maximum measured relief (along one scarp) averages 0.59 km (standard deviation of 0.13 km), with a median of 0.56 km. Additionally, the scarps often terminate at a neighboring scarp, in six cases such that the two scarps are tangent to each other, and in four cases such that they intersect at an angle of ≥45°. These similarities and relationships suggest that the 22 features may be tectonically linked and may have therefore formed as an assemblage within a relatively short interval of time. This assemblage of faults is located in an area of apparently limited lateral variation in crustal thickness, as indicated by crustal models consistent with long-wavelength topography and gravity. If the limited range in crustal thickness in this area was paralleled by a limited range in mechanical lithosphere thickness, this may have facilitated the formation of an assemblage of linked tectonic features.

  16. Airborne LIDAR and high resolution satellite data for rapid 3D feature extraction

    NASA Astrophysics Data System (ADS)

    Jawak, S. D.; Panditrao, S. N.; Luis, A. J.

    2014-11-01

    This work uses the canopy height model (CHM) based workflow for individual tree crown delineation and 3D feature extraction approach (Overwatch Geospatial's proprietary algorithm) for building feature delineation from high-density light detection and ranging (LiDAR) point cloud data in an urban environment and evaluates its accuracy by using very high-resolution panchromatic (PAN) (spatial) and 8-band (multispectral) WorldView-2 (WV-2) imagery. LiDAR point cloud data over San Francisco, California, USA, recorded in June 2010, was used to detect tree and building features by classifying point elevation values. The workflow employed includes resampling of LiDAR point cloud to generate a raster surface or digital terrain model (DTM), generation of a hill-shade image and an intensity image, extraction of digital surface model, generation of bare earth digital elevation model (DEM) and extraction of tree and building features. First, the optical WV-2 data and the LiDAR intensity image were co-registered using ground control points (GCPs). The WV-2 rational polynomial coefficients model (RPC) was executed in ERDAS Leica Photogrammetry Suite (LPS) using supplementary *.RPB file. In the second stage, ortho-rectification was carried out using ERDAS LPS by incorporating well-distributed GCPs. The root mean square error (RMSE) for the WV-2 was estimated to be 0.25 m by using more than 10 well-distributed GCPs. In the second stage, we generated the bare earth DEM from LiDAR point cloud data. In most of the cases, bare earth DEM does not represent true ground elevation. Hence, the model was edited to get the most accurate DEM/ DTM possible and normalized the LiDAR point cloud data based on DTM in order to reduce the effect of undulating terrain. We normalized the vegetation point cloud values by subtracting the ground points (DEM) from the LiDAR point cloud. A normalized digital surface model (nDSM) or CHM was calculated from the LiDAR data by subtracting the DEM from the DSM. The CHM or the normalized DSM represents the absolute height of all aboveground urban features relative to the ground. After normalization, the elevation value of a point indicates the height from the ground to the point. The above-ground points were used for tree feature and building footprint extraction. In individual tree extraction, first and last return point clouds were used along with the bare earth and building footprint models discussed above. In this study, scene dependent extraction criteria were employed to improve the 3D feature extraction process. LiDAR-based refining/ filtering techniques used for bare earth layer extraction were crucial for improving the subsequent 3D features (tree and building) feature extraction. The PAN-sharpened WV-2 image (with 0.5 m spatial resolution) was used to assess the accuracy of LiDAR-based 3D feature extraction. Our analysis provided an accuracy of 98 % for tree feature extraction and 96 % for building feature extraction from LiDAR data. This study could extract total of 15143 tree features using CHM method, out of which total of 14841 were visually interpreted on PAN-sharpened WV-2 image data. The extracted tree features included both shadowed (total 13830) and non-shadowed (total 1011). We note that CHM method could overestimate total of 302 tree features, which were not observed on the WV-2 image. One of the potential sources for tree feature overestimation was observed in case of those tree features which were adjacent to buildings. In case of building feature extraction, the algorithm could extract total of 6117 building features which were interpreted on WV-2 image, even capturing buildings under the trees (total 605) and buildings under shadow (total 112). Overestimation of tree and building features was observed to be limiting factor in 3D feature extraction process. This is due to the incorrect filtering of point cloud in these areas. One of the potential sources of overestimation was the man-made structures, including skyscrapers and bridges, which were confounded and extracted as buildings. This can be attributed to low point density at building edges and on flat roofs or occlusions due to which LiDAR cannot give as much precise planimetric accuracy as photogrammetric techniques (in segmentation) and lack of optimum use of textural information as well as contextual information (especially at walls which are away from roof) in automatic extraction algorithm. In addition, there were no separate classes for bridges or the features lying inside the water and multiple water height levels were also not considered. Based on these inferences, we conclude that the LiDAR-based 3D feature extraction supplemented by high resolution satellite data is a potential application which can be used for understanding and characterization of urban setup.

  17. Sub-population analysis based on temporal features of high content images

    PubMed Central

    2009-01-01

    Background High content screening techniques are increasingly used to understand the regulation and progression of cell motility. The demand of new platforms, coupled with availability of terabytes of data has challenged the traditional technique of identifying cell populations by manual methods and resulted in development of high-dimensional analytical methods. Results In this paper, we present sub-populations analysis of cells at the tissue level by using dynamic features of the cells. We used active contour without edges for segmentation of cells, which preserves the cell morphology, and autoregressive modeling to model cell trajectories. The sub-populations were obtained by clustering static, dynamic and a combination of both features. We were able to identify three unique sub-populations in combined clustering. Conclusion We report a novel method to identify sub-populations using kinetic features and demonstrate that these features improve sub-population analysis at the tissue level. These advances will facilitate the application of high content screening data analysis to new and complex biological problems. PMID:19958514

  18. High order beam features and fitting quadrupole scan data to particle code model.

    SciTech Connect

    Lysenko, W. P.; Garnett, R. W.; Gilpatrick, J. D.; Qiang, J.; Rybarcyk, L. J.; Ryne, Robert; Schneider, J. D.; Smith, H. V.; Young, L. M.; Schulze, M. E.

    2003-01-01

    Quadrupole scans in the HEBT of the 6.7 MeV LEDA RFQ were analyzed to characterize the RFQ output beam. In previous work, profiles measured by the wire scanner were fit to models (beam parameterizations and HEBT simulations) to determine the transverse Courant-Snyder parameters {alpha}, {beta}, and {epsilon} at the RFQ exit. Unfortunately, at the larger quadrupole settings, the measured profiles showed features that were not present in any of our simulations. Here we describe our latest analysis, which resulted in very good fits by using an improved model for the RFQ output beam. The model beam was generated by the RFQ simulation code TOUTATIS. In our fitting code, this beam was distorted by linear transformations that changed the Courant-Snyder parameters to whatever values were required by the nonlinear optimizer while preserving the high-order features of the phase-space distribution. No new physics in the HEBT was required to explain our quad-scan results, just an improved initial beam. High-order features in the RFQ output beam apparently make a significant difference in behavior downstream of the RFQ. While this result gives us increased confidence in our codes, we still have a mystery: exactly what high-order features in the beam are responsible for the the strange behavior downstream. Understanding this phenomenon may be helpful to understanding our halo-experiment data. We have begun to study this by comparing higher-order moments of the TOUTATIS distribution with other distributions.

  19. Extracting features buried within high density atom probe point cloud data through simplicial homology.

    PubMed

    Srinivasan, Srikant; Kaluskar, Kaustubh; Broderick, Scott; Rajan, Krishna

    2015-12-01

    Feature extraction from Atom Probe Tomography (APT) data is usually performed by repeatedly delineating iso-concentration surfaces of a chemical component of the sample material at different values of concentration threshold, until the user visually determines a satisfactory result in line with prior knowledge. However, this approach allows for important features, buried within the sample, to be visually obscured by the high density and volume (~10(7) atoms) of APT data. This work provides a data driven methodology to objectively determine the appropriate concentration threshold for classifying different phases, such as precipitates, by mapping the topology of the APT data set using a concept from algebraic topology termed persistent simplicial homology. A case study of Sc precipitates in an Al-Mg-Sc alloy is presented demonstrating the power of this technique to capture features, such as precise demarcation of Sc clusters and Al segregation at the cluster boundaries, not easily available by routine visual adjustment. PMID:25959554

  20. A comprehensive analysis of earthquake damage patterns using high dimensional model representation feature selection

    NASA Astrophysics Data System (ADS)

    Taşkin Kaya, Gülşen

    2013-10-01

    Recently, earthquake damage assessment using satellite images has been a very popular ongoing research direction. Especially with the availability of very high resolution (VHR) satellite images, a quite detailed damage map based on building scale has been produced, and various studies have also been conducted in the literature. As the spatial resolution of satellite images increases, distinguishability of damage patterns becomes more cruel especially in case of using only the spectral information during classification. In order to overcome this difficulty, textural information needs to be involved to the classification to improve the visual quality and reliability of damage map. There are many kinds of textural information which can be derived from VHR satellite images depending on the algorithm used. However, extraction of textural information and evaluation of them have been generally a time consuming process especially for the large areas affected from the earthquake due to the size of VHR image. Therefore, in order to provide a quick damage map, the most useful features describing damage patterns needs to be known in advance as well as the redundant features. In this study, a very high resolution satellite image after Iran, Bam earthquake was used to identify the earthquake damage. Not only the spectral information, textural information was also used during the classification. For textural information, second order Haralick features were extracted from the panchromatic image for the area of interest using gray level co-occurrence matrix with different size of windows and directions. In addition to using spatial features in classification, the most useful features representing the damage characteristic were selected with a novel feature selection method based on high dimensional model representation (HDMR) giving sensitivity of each feature during classification. The method called HDMR was recently proposed as an efficient tool to capture the input-output relationships in high-dimensional systems for many problems in science and engineering. The HDMR method is developed to improve the efficiency of the deducing high dimensional behaviors. The method is formed by a particular organization of low dimensional component functions, in which each function is the contribution of one or more input variables to the output variables.

  1. Uranus' Persistent Patterns and Features from High-SNR Imaging in 2012-2014

    NASA Astrophysics Data System (ADS)

    Fry, Patrick M.; Sromovsky, Lawrence A.; de Pater, Imke; Hammel, Heidi B.; Marcus, Phillip

    2015-11-01

    Since 2012, Uranus has been the subject of an observing campaign utilizing high signal-to-noise imaging techniques at Keck Observatory (Fry et al. 2012, Astron. J. 143, 150-161). High quality observing conditions on four observing runs of consecutive nights allowed longitudinally-complete coverage of the atmosphere over a period of two years (Sromovsky et al. 2015, Icarus 258, 192-223). Global mosaic maps made from images acquired on successive nights in August 2012, November 2012, August 2013, and August 2014, show persistent patterns, and six easily distinguished long-lived cloud features, which we were able to track for long periods that ranged from 5 months to over two years. Two at similar latitudes are associated with dark spots, and move with the atmospheric zonal flow close to the location of their associated dark spot instead of following the flow at the latitude of the bright features. These features retained their morphologies and drift rates in spite of several close interactions. A second pair of features at similar latitudes also survived several close approaches. Several of the long-lived features also exhibited equatorward drifts and latitudinal oscillations. Also persistent are a remarkable near-equatorial wave feature and global zonal band structure. We will present imagery, maps, and analyses of these phenomena.PMF and LAS acknowledge support from NASA Planetary Astronomy Program; PMF and LAS acknowledge funding and technical support from W. M. Keck Observatory. We thank those of Hawaiian ancestry on whose sacred mountain we are privileged to be guests. Without their generous hospitality none of our groundbased observations would have been possible.

  2. A PLANETARY LENSING FEATURE IN CAUSTIC-CROSSING HIGH-MAGNIFICATION MICROLENSING EVENTS

    SciTech Connect

    Chung, Sun-Ju; Hwang, Kyu-Ha; Ryu, Yoon-Hyun; Lee, Chung-Uk E-mail: kyuha@kasi.re.kr E-mail: leecu@kasi.re.kr

    2012-05-20

    Current microlensing follow-up observations focus on high-magnification events because of the high efficiency of planet detection. However, central perturbations of high-magnification events caused by a planet can also be produced by a very close or a very wide binary companion, and the two kinds of central perturbations are not generally distinguished without time consuming detailed modeling (a planet-binary degeneracy). Hence, it is important to resolve the planet-binary degeneracy that occurs in high-magnification events. In this paper, we investigate caustic-crossing high-magnification events caused by a planet and a wide binary companion. From this investigation, we find that because of the different magnification excess patterns inside the central caustics induced by the planet and the binary companion, the light curves of the caustic-crossing planetary-lensing events exhibit a feature that is discriminated from those of the caustic-crossing binary-lensing events, and the feature can be used to immediately distinguish between the planetary and binary companions. The planetary-lensing feature appears in the interpeak region between the two peaks of the caustic-crossings. The structure of the interpeak region for the planetary-lensing events is smooth and convex or boxy, whereas the structure for the binary-lensing events is smooth and concave. We also investigate the effect of a finite background source star on the planetary-lensing feature in the caustic-crossing high-magnification events. From this, we find that the convex-shaped interpeak structure appears in a certain range that changes with the mass ratio of the planet to the planet-hosting star.

  3. Prognostic differences of World Health Organization-assessed mitotic activity index and mitotic impression by quick scanning in invasive ductal breast cancer patients younger than 55 years.

    PubMed

    Skaland, Ivar; van Diest, Paul J; Janssen, Emiel A M; Gudlaugsson, Einar; Baak, Jan P A

    2008-04-01

    The proliferation marker mitotic activity index is the strongest prognostic indicator in lymph node-negative breast cancer. The World Health Organization (WHO) 2003-defined procedure for determining WHO-mitotic activity index is often replaced by a quick scan mitotic impression. We evaluated the prognostic consequences of this practice in 433 T(1-3)N(0)M(0) lymph node-negative invasive ductal type breast cancers with long-term follow-up (median, 112 months; range, 12-187 months). Twenty-seven percent of the studied cases developed distant metastases, and 25% died of disease. Agreement between WHO-mitotic activity index (0-5 = 1, 6-10 = 2, >10 = 3) and mitotic impression (1, 2, 3) categories was 66% (kappa = 0.41), including 85% for category 1, 26% for category 2, and 52% for category 3. The WHO-mitotic activity index was a much stronger prognosticator than the mitotic impression, and the 10-year survival rates of the same categories (eg, mitotic activity index and mitotic impression category both 2) differed greatly. When grade was assessed by combining WHO-mitotic activity index or mitotic impression with the same values for tubular formation and nuclear atypia, grades disagreed in 18% of the cases. Deviation from the formal WHO-mitotic activity index assessment guidelines in breast cancer often results in erroneous prognosis estimations with therapeutic consequences and may explain why the prognostic value of proliferative activity in breast cancer is not always confirmed. PMID:18291440

  4. Common variants spanning PLK4 are associated with mitotic-origin aneuploidy in human embryos.

    PubMed

    McCoy, Rajiv C; Demko, Zachary; Ryan, Allison; Banjevic, Milena; Hill, Matthew; Sigurjonsson, Styrmir; Rabinowitz, Matthew; Fraser, Hunter B; Petrov, Dmitri A

    2015-04-10

    Aneuploidy, the inheritance of an atypical chromosome complement, is common in early human development and is the primary cause of pregnancy loss. By screening day-3 embryos during in vitro fertilization cycles, we identified an association between aneuploidy of putative mitotic origin and linked genetic variants on chromosome 4 of maternal genomes. This associated region contains a candidate gene, Polo-like kinase 4 (PLK4), that plays a well-characterized role in centriole duplication and has the ability to alter mitotic fidelity upon minor dysregulation. Mothers with the high-risk genotypes contributed fewer embryos for testing at day 5, suggesting that their embryos are less likely to survive to blastocyst formation. The associated region coincides with a signature of a selective sweep in ancient humans, suggesting that the causal variant was either the target of selection or hitchhiked to substantial frequency. PMID:25859044

  5. Mcl-1 dynamics influence mitotic slippage and death in mitosis

    PubMed Central

    Sloss, Olivia; Topham, Caroline; Diez, Maria; Taylor, Stephen

    2016-01-01

    Microtubule-binding drugs such as taxol are frontline treatments for a variety of cancers but exactly how they yield patient benefit is unclear. In cell culture, inhibiting microtubule dynamics prevents spindle assembly, leading to mitotic arrest followed by either apoptosis in mitosis or slippage, whereby a cell returns to interphase without dividing. Myeloid cell leukaemia-1 (Mcl-1), a pro-survival member of the Bcl-2 family central to the intrinsic apoptosis pathway, is degraded during a prolonged mitotic arrest and may therefore act as a mitotic death timer. Consistently, we show that blocking proteasome-mediated degradation inhibits taxol-induced mitotic apoptosis in a Mcl-1-dependent manner. However, this degradation does not require the activity of either APC/C-Cdc20, FBW7 or MULE, three separate E3 ubiquitin ligases implicated in targeting Mcl-1 for degradation. This therefore challenges the notion that Mcl-1 undergoes regulated degradation during mitosis. We also show that Mcl-1 is continuously synthesized during mitosis and that blocking protein synthesis accelerates taxol induced death-in-mitosis. Modulating Mcl-1 levels also influences slippage; overexpressing Mcl-1 extends the time from mitotic entry to mitotic exit in the presence of taxol, while inhibiting Mcl-1 accelerates it. We suggest that Mcl-1 competes with Cyclin B1 for binding to components of the proteolysis machinery, thereby slowing down the slow degradation of Cyclin B1 responsible for slippage. Thus, modulating Mcl-1 dynamics influences both death-in-mitosis and slippage. However, because mitotic degradation of Mcl-1 appears not to be under the control of an E3 ligase, we suggest that the notion of network crosstalk is used with caution. PMID:26769847

  6. PKCι depletion initiates mitotic slippage-induced senescence in glioblastoma

    PubMed Central

    Restall, Ian J; Parolin, Doris A E; Daneshmand, Manijeh; Hanson, Jennifer E L; Simard, Manon A; Fitzpatrick, Megan E; Kumar, Ritesh; Lavictoire, Sylvie J; Lorimer, Ian A J

    2015-01-01

    Cellular senescence is a tumor suppressor mechanism where cells enter a permanent growth arrest following cellular stress. Oncogene-induced senescence (OIS) is induced in non-malignant cells following the expression of an oncogene or inactivation of a tumor suppressor. Previously, we have shown that protein kinase C iota (PKCι) depletion induces cellular senescence in glioblastoma cells in the absence of a detectable DNA damage response. Here we demonstrate that senescent glioblastoma cells exhibit an aberrant centrosome morphology. This was observed in basal levels of senescence, in p21-induced senescence, and in PKCι depletion-induced senescence. In addition, senescent glioblastoma cells are polyploid, Ki-67 negative and arrest at the G1/S checkpoint, as determined by expression of cell cycle regulatory proteins. These markers are all consistent with cells that have undergone mitotic slippage. Failure of the spindle assembly checkpoint to function properly can lead to mitotic slippage, resulting in the premature exit of mitotic cells into the G1 phase of the cell cycle. Although in G1, these cells have the replicated DNA and centrosomal phenotype of a cell that has entered mitosis and failed to divide. Overall, we demonstrate that PKCι depletion initiates mitotic slippage-induced senescence in glioblastoma cells. To our knowledge, this is the first evidence of markers of mitotic slippage directly in senescent cells by co-staining for senescence-associated β-galactosidase and immunofluorescence markers in the same cell population. We suggest that markers of mitotic slippage be assessed in future studies of senescence to determine the extent of mitotic slippage in the induction of cellular senescence. PMID:26208522

  7. Picropodophyllin causes mitotic arrest and catastrophe by depolymerizing microtubules via Insulin-like growth factor-1 receptor-independent mechanism

    PubMed Central

    Waraky, Ahmed; Akopyan, Karen; Parrow, Vendela; Strömberg, Thomas; Axelson, Magnus; Abrahmsén, Lars; Lindqvist, Arne; Larsson, Olle; Aleem, Eiman

    2014-01-01

    Picropodophyllin (PPP) is an anticancer drug undergoing clinical development in NSCLC. PPP has been shown to suppress IGF-1R signaling and to induce a G2/M cell cycle phase arrest but the exact mechanisms remain to be elucidated. The present study identified an IGF-1-independent mechanism of PPP leading to pro-metaphase arrest. The mitotic block was induced in human cancer cell lines and in an A549 xenograft mouse but did not occur in normal hepatocytes/mouse tissues. Cell cycle arrest by PPP occurred in vitro and in vivo accompanied by prominent CDK1 activation, and was IGF-1R-independent since it occurred also in IGF-1R-depleted and null cells. The tumor cells were not arrested in G2/M but in mitosis. Centrosome separation was prevented during mitotic entry, resulting in a monopolar mitotic spindle with subsequent prometaphase-arrest, independent of Plk1/Aurora A or Eg5, and leading to cell features of mitotic catastrophe. PPP also increased soluble tubulin and decreased spindle-associated tubulin within minutes, indicating that it interfered with microtubule dynamics. These results provide a novel IGF-1R-independent mechanism of antitumor effects of PPP. PMID:25268741

  8. Cold-treated centrosome: isolation of centrosomes from mitotic sea urchin eggs, production of an anticentrosomal antibody, and novel ultrastructural imaging.

    PubMed

    Thompson-Coffe, C; Coffe, G; Schatten, H; Mazia, D; Schatten, G

    1996-01-01

    A novel isolation of centrosomes is described and it was used to both generate a centrosome-specific monoclonal antibody and to image with high-resolution low-voltage scanning electron microscopy the surface details of the isolated centrosome. At first mitotic prometaphase, sea urchin zygotes are chilled on ice overnight. While most of the microtubules disassemble, the mitotic centrosomes collapse into aggregated masses. These centrosomes have been isolated, and used to generate a monoclonal antibody, designated 4D2, which is reactive with interphase and mitotic centrosomes. 4D2 staining of centrosomes is similar, but not identical, to that of other centrosomal antibodies like Ah6 and 5051. Centrosomal material is detected as a compact sphere after cold treatment; upon recovery the sphere expands and undergoes the shape changes previously described [Mazia et al., 1987: J. Cell Biol. 105:206a] to eventually reorganize a normal mitotic apparatus. PMID:8674139

  9. Mutagenic analysis of the destruction signal of mitotic cyclins and structural characterization of ubiquitinated intermediates.

    PubMed Central

    King, R W; Glotzer, M; Kirschner, M W

    1996-01-01

    Mitotic cyclins are abruptly degraded at the end of mitosis by a cell-cycle-regulated ubiquitin-dependent proteolytic system. To understand how cyclin is recognized for ubiquitin conjugation, we have performed a mutagenic analysis of the destruction signal of mitotic cyclins. We demonstrate that an N-terminal cyclin B segment as short as 27 residues, containing the 9-amino-acid destruction box, is sufficient to destabilize a heterologous protein in mitotic Xenopus extracts. Each of the three highly conserved residues of the cyclin B destruction box is essential for ubiquitination and subsequent degradation. Although an intact destruction box is essential for the degradation of both A- and B-type cyclins, we find that the Xenopus cyclin A1 destruction box cannot functionally substitute for its B-type counterpart, because it does not contain the highly conserved asparagine necessary for cyclin B proteolysis. Physical analysis of ubiquitinated cyclin B intermediates demonstrates that multiple lysine residues function as ubiquitin acceptor sites, and mutagenic studies indicate that no single lysine residue is essential for cyclin B degradation. This study defines the key residues of the destruction box that target cyclin for ubiquitination and suggests there are important differences in the way in which A- and B-type cyclins are recognized by the cyclin ubiquitination machinery. Images PMID:8885231

  10. Dyskerin Localizes to the Mitotic Apparatus and Is Required for Orderly Mitosis in Human Cells

    PubMed Central

    Alawi, Faizan; Lin, Ping

    2013-01-01

    Dyskerin is a highly conserved, nucleolar RNA-binding protein with established roles in small nuclear ribonucleoprotein biogenesis, telomerase and telomere maintenance and precursor rRNA processing. Telomerase is functional during S phase and the bulk of rRNA maturation occurs during G1 and S phases; both processes are inactivated during mitosis. Yet, we show that during the course of cell cycle progression, human dyskerin expression peaks during G2/M in parallel with the upregulation of pro-mitotic factors. Dyskerin redistributed from the nucleolus in interphase cells to the perichromosomal region during prometaphase, metaphase and anaphase. With continued anaphase progression, dyskerin also localized to the cytoplasm within the mid-pole region. Loss of dyskerin function via siRNA-mediated depletion promoted G2/M accumulation and this was accompanied by an increased mitotic index and activation of the spindle assembly checkpoint. Live cell imaging further revealed an array of mitotic defects including delayed prometaphase progression, a significantly increased incidence of multi-polar spindles, and anaphase bridges culminating in micronucleus formation. Together, these findings suggest that dyskerin is a highly dynamic protein throughout the cell cycle and increases the repertoire of fundamental cellular processes that are disrupted by absence of its normal function. PMID:24303026

  11. High-Velocity Absorption Features in FUSE Spectra of Eta Carinae

    NASA Technical Reports Server (NTRS)

    Sonneborn, G.; Iping, R. C.; Gull, T. R.; Vieira, G.

    2003-01-01

    Numerous broad (200 to 1000 km/sec) features in the FUSE spectrum (905-1187 A) of eta Carinae are identified as absorption by a forest of high-velocity narrow lines formed in the expanding circumstellar envelope. These features were previously thought to be P-Cygni lines arising in the wind of the central star. The features span a heliocentric velocity range of -140 to -580 km/sec and are seen prominently in low-ionization ground-state transitions (e.g. N I 1134-35, Fe II 1145-42, 1133, 1127- 22, P II 1153, C I 1158) in addition to C III] 1176 A. The high-velocity components of the FUSE transitions have depths about 50% below the continuum. The identifications are consistent with the complex velocity structures seen in ground- and excited-state transitions of Mg I, Mg 11, Fe II, V II, etc observed in STIS/E230H spectra. The origin of other broad features of similar width and depth in the FUSE spectrum, but without low-velocity ISM absorption, are unidentified. However, they are suspected of being absorption of singly-ionized iron-peak elements (e.g. Fe II, V II, Cr II) out of excited levels 1,000 to 20,000 cmE-l above the ground state. The high-velocity features seen in Fe II 1145 are also present in Fe II 1608 (STIS/E140M), but are highly saturated in the latter. Since these transitions have nearly identical log (flambda) (1.998 vs. 2.080), the differences in the profiles are attributable to the different aperture sizes used (30 x 30 arcsec for FUSE, 0.2 x 0.2 arcsec for STIS/E140M). The high-velocity gas appears to be very patchy or has a small covering factor near the central star. Eta Carinae has been observed several times by FUSE over the past three years. The FUSE flux levels and spectral features in eta Car are essentially unchanged over the 2000 March to June 2002 period, establishing a baseline far-UV spectrum in advance of the predicted spectroscopic minimum in 2003.

  12. Mitotic catastrophe and cell death induced by depletion of centrosomal proteins

    PubMed Central

    Kimura, M; Yoshioka, T; Saio, M; Banno, Y; Nagaoka, H; Okano, Y

    2013-01-01

    Mitotic catastrophe, which refers to cell death or its prologue triggered by aberrant mitosis, can be induced by a heterogeneous group of stimuli, including chromosome damage or perturbation of the mitotic apparatus. We investigated the mechanism of mitotic catastrophe and cell death induced by depletion of centrosomal proteins that perturbs microtubule organization. We transfected cells harboring wild-type or mutated p53 with siRNAs targeting Aurora A, ninein, TOG, TACC3, γ-tubulin, or pericentriolar material-1, and monitored the effects on cell death. Knockdown of Aurora A, ninein, TOG, and TACC3 led to cell death, regardless of p53 status. Knockdown of Aurora A, ninein, and TOG, led to aberrant spindle formation and subsequent cell death, which was accompanied by several features of apoptosis, including nuclear condensation and Annexin V binding in HeLa cells. During this process, cleavage of poly(ADP-ribose) polymerase-1, caspase-3, and caspase-9 was detected, but cleavage of caspase-8 was not. Cell death, monitored by time-lapse imaging, occurred during both interphase and M phase. In cells depleted of a centrosomal protein (Aurora A, ninein, or TOG), the rate of cell death was higher if the cells were cotransfected with siRNA against BubR1 or Mad2 than if they were transfected with siRNA against Bub1 or a control siRNA. These results suggest that metaphase arrest is necessary for the mitotic catastrophe and cell death caused by depletion of centrosomal proteins. Knockdown of centrosomal proteins led to increased phosphorylation of Chk2. Enhanced p-Chk2 localization was also observed at the centrosome in cells arrested in M phase, as well as in the nuclei of dying cells. Cotransfection of siRNAs against Chk2, in combination with depletion of a centrosomal protein, decreased the amount of cell death. Thus, Chk2 activity is indispensable for apoptosis after mitotic catastrophe induced by depletion of centrosomal proteins that perturbs microtubule organization. PMID:23598415

  13. Promotion of chloroplast proliferation upon enhanced post-mitotic cell expansion in leaves

    PubMed Central

    2013-01-01

    Background Leaves are determinate organs; hence, precise control of cell proliferation and post-mitotic cell expansion is essential for their growth. A defect in cell proliferation often triggers enhanced post-mitotic cell expansion in leaves. This phenomenon is referred to as ‘compensation’. Several lines of evidence from studies on compensation have shown that cell proliferation and post-mitotic cell expansion are coordinately regulated during leaf development. Therefore, compensation has attracted much attention to the mechanisms for leaf growth. However, our understanding of compensation at the subcellular level remains limited because studies of compensation have focused mainly on cellular-level phenotypes. Proper leaf growth requires quantitative control of subcellular components in association with cellular-level changes. To gain insight into the subcellular aspect of compensation, we investigated the well-known relationship between cell area and chloroplast number per cell in compensation-exhibiting lines, and asked whether chloroplast proliferation is modulated in response to the induction of compensation. Results We first established a convenient and reliable method for observation of chloroplasts in situ. Using this method, we analyzed Arabidopsis thaliana mutants fugu5 and angustifolia3 (an3), and a transgenic line KIP-RELATED PROTEIN2 overexpressor (KRP2 OE), which are known to exhibit typical features of compensation. We here showed that chloroplast number per cell increased in the subepidermal palisade tissue of these lines. We analyzed tetraploidized wild type, fugu5, an3 and KRP2 OE, and found that cell area itself, but not nuclear ploidy, is a key parameter that determines the activity of chloroplast proliferation. In particular, in the case of an3, we uncovered that promotion of chloroplast proliferation depends on the enhanced post-mitotic cell expansion. The expression levels of chloroplast proliferation-related genes are similar to or lower than that in the wild type during this process. Conclusions This study demonstrates that chloroplast proliferation is promoted in compensation-exhibiting lines. This promotion of chloroplast proliferation takes place in response to cell-area increase in post-mitotic phase in an3. The expression of chloroplast proliferation-related genes were not promoted in compensation-exhibiting lines including an3, arguing that an as-yet-unknown mechanism is responsible for modulation of chloroplast proliferation in these lines. PMID:24074400

  14. Rabbit System. Low cost, high reliability front end electronics featuring 16 bit dynamic range

    NASA Astrophysics Data System (ADS)

    Drake, G.; Droege, T. F.; Nelson, C. A., Jr.; Turner, K. J.; Ohska, T. K.

    1985-10-01

    A new crate-based front end system has been built which features low cost, compact packaging, command capability, 16 bit dynamic range digitization, and a high degree of redundancy. The crate can contain a variety of instrumentation modules, and is designed to be situated close to the detector. The system is suitable for readout of a large number of channels via parallel multiprocessor data acquisition.

  15. New features in Saturn's atmosphere revealed by high-resolution thermal infrared images

    NASA Technical Reports Server (NTRS)

    Gezari, D. Y.; Mumma, M. J.; Espenak, F.; Deming, D.; Bjoraker, G.; Woods, L.; Folz, W.

    1989-01-01

    Observations of the stratospheric IR emission structure on Saturn are presented. The high-spatial-resolution global images show a variety of new features, including a narrow equatorial belt of enhanced emission at 7.8 micron, a prominent symmetrical north polar hotspot at all three wavelengths, and a midlatitude structure which is asymmetrically brightened at the east limb. The results confirm the polar brightening and reversal in position predicted by recent models for seasonal thermal variations of Saturn's stratosphere.

  16. Optimal strategies for sequential validation of significant features from high-dimensional genomic data.

    PubMed

    Lohr, Miriam; Köllmann, Claudia; Freis, Evgenia; Hellwig, Birte; Hengstler, Jan G; Ickstadt, Katja; Rahnenführer, Jörg

    2012-01-01

    High-dimensional genomic studies play a key role in identifying critical features that are significantly associated with a phenotypic outcome. The two most important examples are the detection of (1) differentially expressed genes from genome-wide gene expression studies and (2) single-nucleotide polymorphisms (SNPs) from genome-wide association studies. Such experiments are often associated with high noise levels, and the validity of statistical conclusions suffers from low sample size compared to large number of features. The corresponding multiple testing problem calls for the identification of optimal strategies for controlling the numbers of false discoveries and false nondiscoveries. In addition, a frequent validation problem is that features identified as important in one study are often less so in another study. Adjustment for multiple testing in both studies separately increases the risk of missing the crucial features even further. These problems can be addressed by sequential validation strategies, where only significant features identified in one study enter as candidates in the next study. The quality associated with different studies, for example, in terms of noise levels, may vary considerably. By performing simulation studies it is possible to demonstrate that the optimal order for this stepwise procedure is to sort experimental studies according to their quality in descending order. The impact of the method for multiple testing adjustment (Bonferroni-Holm, FDR) was also analyzed. Finally, the sequential validation strategy was applied to three large breast cancer studies with gene expression measurements, confirming the crucial impact of the order of the validation steps in a real-world application. PMID:22686304

  17. Mitotic Chromosome Loss in a Disomic Haploid of SACCHAROMYCES CEREVISIAE

    PubMed Central

    Campbell, D. A.; Fogel, S.; Lusnak, K.

    1975-01-01

    Experiments designed to characterize the incidence of mitotic chromosome loss in a yeast disomic haploid were performed. The selective methods employed utilize the non-mating property of strains disomic for linkage group III and heterozygous at the mating type locus. The principal findings are: (1) The frequency of spontaneous chromosome loss in the disome is of the order 10-4 per cell; this value approximates the frequency in the same population of spontaneous mitotic exchange resulting in homozygosity at the mating type locus. (2) The recovered diploids are pure clones, and thus represent unique events in the disomic haploid. (3) Of the euploid chromosomes recovered after events leading to chromosome loss, approximately 90% retain the parental marker configuration expected from segregation alone; however, the remainder are recombinant for marker genes, and are the result of mitotic exchanges in the disome, especially in regions near the centromere. The recombinant proportion significantly exceeds that expected if chromosome loss and mitotic exchange in the disome were independent events. The data are consistent with a model proposing mitotic nondisjunction as the event responsible for chromosome loss in the disomic haploid. PMID:1092597

  18. Mechanical control of mitotic progression in single animal cells

    PubMed Central

    Cattin, Cedric J.; Düggelin, Marcel; Martinez-Martin, David; Gerber, Christoph; Müller, Daniel J.; Stewart, Martin P.

    2015-01-01

    Despite the importance of mitotic cell rounding in tissue development and cell proliferation, there remains a paucity of approaches to investigate the mechanical robustness of cell rounding. Here we introduce ion beam-sculpted microcantilevers that enable precise force-feedback–controlled confinement of single cells while characterizing their progression through mitosis. We identify three force regimes according to the cell response: small forces (∼5 nN) that accelerate mitotic progression, intermediate forces where cells resist confinement (50–100 nN), and yield forces (>100 nN) where a significant decline in cell height impinges on microtubule spindle function, thereby inhibiting mitotic progression. Yield forces are coincident with a nonlinear drop in cell height potentiated by persistent blebbing and loss of cortical F-actin homogeneity. Our results suggest that a buildup of actomyosin-dependent cortical tension and intracellular pressure precedes mechanical failure, or herniation, of the cell cortex at the yield force. Thus, we reveal how the mechanical properties of mitotic cells and their response to external forces are linked to mitotic progression under conditions of mechanical confinement. PMID:26305930

  19. Fabrication of Pt nanowires with a diffraction-unlimited feature size by high-threshold lithography

    NASA Astrophysics Data System (ADS)

    Li, Li; Wang, Zuobin; Li, Wenjun; Peng, Kuiqing; Zhang, Ziang; Yu, Miao; Song, Zhengxun; Weng, Zhankun; Wang, Dapeng; Zhao, Le

    2015-09-01

    Although the nanoscale world can already be observed at a diffraction-unlimited resolution using far-field optical microscopy, to make the step from microscopy to lithography still requires a suitable photoresist material system. In this letter, we consider the threshold to be a region with a width characterized by the extreme feature size obtained using a Gaussian beam spot. By narrowing such a region through improvement of the threshold sensitization to intensity in a high-threshold material system, the minimal feature size becomes smaller. By using platinum as the negative photoresist, we demonstrate that high-threshold lithography can be used to fabricate nanowire arrays with a scalable resolution along the axial direction of the linewidth from the micro- to the nanoscale using a nanosecond-pulsed laser source with a wavelength λ0 = 1064 nm. The minimal feature size is only several nanometers (sub λ0/100). Compared with conventional polymer resist lithography, the advantages of high-threshold lithography are sharper pinpoints of laser intensity triggering the threshold response and also higher robustness allowing for large area exposure by a less-expensive nanosecond-pulsed laser.

  20. Unbiased Prediction and Feature Selection in High-Dimensional Survival Regression

    PubMed Central

    Laimighofer, Michael; Krumsiek, Jan; Theis, Fabian J.

    2016-01-01

    Abstract With widespread availability of omics profiling techniques, the analysis and interpretation of high-dimensional omics data, for example, for biomarkers, is becoming an increasingly important part of clinical medicine because such datasets constitute a promising resource for predicting survival outcomes. However, early experience has shown that biomarkers often generalize poorly. Thus, it is crucial that models are not overfitted and give accurate results with new data. In addition, reliable detection of multivariate biomarkers with high predictive power (feature selection) is of particular interest in clinical settings. We present an approach that addresses both aspects in high-dimensional survival models. Within a nested cross-validation (CV), we fit a survival model, evaluate a dataset in an unbiased fashion, and select features with the best predictive power by applying a weighted combination of CV runs. We evaluate our approach using simulated toy data, as well as three breast cancer datasets, to predict the survival of breast cancer patients after treatment. In all datasets, we achieve more reliable estimation of predictive power for unseen cases and better predictive performance compared to the standard CoxLasso model. Taken together, we present a comprehensive and flexible framework for survival models, including performance estimation, final feature selection, and final model construction. The proposed algorithm is implemented in an open source R package (SurvRank) available on CRAN. PMID:26894327

  1. Glacial Features in the Western Gulf of Maine Inferred From High Resolution Bathymetric Data

    NASA Astrophysics Data System (ADS)

    Malik, M. A.; Licciardi, J. M.; Ward, L. G.; Mayer, L. A.

    2007-12-01

    Multibeam sonar surveys in the last decade have revealed submerged glacial features in the western Gulf of Maine (e.g., Valentine et al., 2003). Here we examine high-resolution multibeam bathymetric data acquired in 2001 and 2005 over Jeffreys Ledge to infer the origin of previously unrecognized small-scale marine glacial features. Ridges as high as 5 m appear throughout the length of Jeffreys Ledge in water depths of ~50 m. Bottom photographs of these features show boulders of up to 50 cm diameter in a flat sandy bottom devoid of finer material. These ridges are probably recessional moraines that have been reworked during lower relative sea level (~55 m below modern sea level). The moraine-like features imply stabilization of an ice margin along the length of Jeffreys Ledge. The central portion of Jeffreys Ledge also contains asymmetrical dune forms with a relief of 1-6 m and along-crest orientations trending NW-SE. These dunes may have formed during megaflood events with water flow toward the southwest. Streamlined bathymetric features with a relief of ~8 m and lengths up to 700 m occur east of Jeffreys Ledge. These features have similar dimensions but different orientations (N-S), as compared to southeast-oriented drumlins identified south of Cape Ann by Oldale et al. (1994). Dissimilar orientations of these drumlins are consistent with the lobate shape of the ice sheet and probable local ice flow directions. Numerous iceberg scours were observed in the basins east of Stellwagen Bank and Jeffreys Ledge with varying widths (50-300 m), scour depths (1-5 m) and lengths (3-10 km). Two dominant orientations of iceberg scours (E- W and N-S) were identified. Additional data such as seismic profiles, bottom photographs and bottom samples will further define the origin of these small-scale glacial features. Oldale, R.N., Knebel, H.J., Bothner, M.H., 1994, Geomorphology 9, 301-309. Valentine, P., Unger, T., Baker, J., 2003, U.S. Geological Survey Geologic Investigations Series Map I-2676C, scale 1:60,000.

  2. Centrin: Another target of monastrol, an inhibitor of mitotic spindle

    NASA Astrophysics Data System (ADS)

    Duan, Lian; Wang, Tong-Qing; Bian, Wei; Liu, Wen; Sun, Yue; Yang, Bin-Sheng

    2015-02-01

    Monastrol, a cell-permeable inhibitor, considered to specifically inhibit kinesin Eg5, can cause mitotic arrest and monopolar spindle formation, thus exhibiting antitumor properties. Centrin, a ubiquitous protein associated with centrosome, plays a critical role in centrosome duplication. Moreover, a correlation between centrosome amplification and cancer has been reported. In this study, it is proposed for the first time that centrin may be another target of the anticancer drug monastrol since monastrol can effectively inhibit not only the growth of the transformed Escherichia coli cells in vivo, but also the Lu3+-dependent self-assembly of EoCen in vitro. The two closely related compounds (Compounds 1 and 2) could not take the same effect. Fluorescence titration experiments suggest that four monastrols per protein is the optimum binding pattern, and the binding constants at different temperatures were obtained. Detailed thermodynamic analysis indicates that hydrophobic force is the main acting force between monastrol and centrin, and the extent of monastrol inhibition of centrin self-assembly is highly dependent upon the hydrophobic region of the protein, which is largely exposed by the binding of metal ions.

  3. Increased functional protein expression using nucleotide sequence features enriched in highly expressed genes in zebrafish.

    PubMed

    Horstick, Eric J; Jordan, Diana C; Bergeron, Sadie A; Tabor, Kathryn M; Serpe, Mihaela; Feldman, Benjamin; Burgess, Harold A

    2015-04-20

    Many genetic manipulations are limited by difficulty in obtaining adequate levels of protein expression. Bioinformatic and experimental studies have identified nucleotide sequence features that may increase expression, however it is difficult to assess the relative influence of these features. Zebrafish embryos are rapidly injected with calibrated doses of mRNA, enabling the effects of multiple sequence changes to be compared in vivo. Using RNAseq and microarray data, we identified a set of genes that are highly expressed in zebrafish embryos and systematically analyzed for enrichment of sequence features correlated with levels of protein expression. We then tested enriched features by embryo microinjection and functional tests of multiple protein reporters. Codon selection, releasing factor recognition sequence and specific introns and 3' untranslated regions each increased protein expression between 1.5- and 3-fold. These results suggested principles for increasing protein yield in zebrafish through biomolecular engineering. We implemented these principles for rational gene design in software for codon selection (CodonZ) and plasmid vectors incorporating the most active non-coding elements. Rational gene design thus significantly boosts expression in zebrafish, and a similar approach will likely elevate expression in other animal models. PMID:25628360

  4. Increased functional protein expression using nucleotide sequence features enriched in highly expressed genes in zebrafish

    PubMed Central

    Horstick, Eric J.; Jordan, Diana C.; Bergeron, Sadie A.; Tabor, Kathryn M.; Serpe, Mihaela; Feldman, Benjamin; Burgess, Harold A.

    2015-01-01

    Many genetic manipulations are limited by difficulty in obtaining adequate levels of protein expression. Bioinformatic and experimental studies have identified nucleotide sequence features that may increase expression, however it is difficult to assess the relative influence of these features. Zebrafish embryos are rapidly injected with calibrated doses of mRNA, enabling the effects of multiple sequence changes to be compared in vivo. Using RNAseq and microarray data, we identified a set of genes that are highly expressed in zebrafish embryos and systematically analyzed for enrichment of sequence features correlated with levels of protein expression. We then tested enriched features by embryo microinjection and functional tests of multiple protein reporters. Codon selection, releasing factor recognition sequence and specific introns and 3′ untranslated regions each increased protein expression between 1.5- and 3-fold. These results suggested principles for increasing protein yield in zebrafish through biomolecular engineering. We implemented these principles for rational gene design in software for codon selection (CodonZ) and plasmid vectors incorporating the most active non-coding elements. Rational gene design thus significantly boosts expression in zebrafish, and a similar approach will likely elevate expression in other animal models. PMID:25628360

  5. p21-activated kinase 4 regulates mitotic spindle positioning and orientation.

    PubMed

    Bompard, Guillaume; Morin, Nathalie

    2012-01-01

    During mitosis, microtubules (MTs) are massively rearranged into three sets of highly dynamic MTs that are nucleated from the centrosomes to form the mitotic spindle. Tight regulation of spindle positioning in the dividing cell and chromosome alignment at the center of the metaphase spindle are required to ensure perfect chromosome segregation and to position the cytokinetic furrow that will specify the two daughter cells. Spindle positioning requires regulation of MT dynamics, involving depolymerase activities together with cortical and kinetochore-mediated pushing and pulling forces acting on astral MTs and kinetochore fibres. These forces rely on MT motor activities. Cortical pulling forces exerted on astral MTs depend upon dynein/dynactin complexes and are essential in both symmetric and asymmetric cell division. A well-established spindle positioning pathway regulating the cortical targeting of dynein/dynactin involves the conserved LGN (Leu-Gly-Asn repeat-enriched-protein) and NuMA (microtubule binding nuclear mitotic apparatus protein) complex. Spindle orientation is also regulated by integrin-mediated cell adhesion and actin retraction fibres that respond to mechanical stress and are influenced by the microenvironment of the dividing cell. Altering the capture of astral MTs or modulating pulling forces affects spindle position, which can impair cell division, differentiation and embryogenesis. In this general scheme, the activity of mitotic kinases such as Auroras and Plk1 (Polo-like kinase 1) is crucial. Recently, the p21-activated kinases (PAKs) emerged as novel important players in mitotic progression. In our recent article, we demonstrated that PAK4 regulates spindle positioning in symmetric cell division. In this commentary, and in light of recent published studies, we discuss how PAK4 could participate in the regulation of mechanisms involved in spindle positioning and orientation. PMID:22960742

  6. p21-activated kinase 4 regulates mitotic spindle positioning and orientation

    PubMed Central

    Bompard, Guillaume; Morin, Nathalie

    2012-01-01

    During mitosis, microtubules (MTs) are massively rearranged into three sets of highly dynamic MTs that are nucleated from the centrosomes to form the mitotic spindle. Tight regulation of spindle positioning in the dividing cell and chromosome alignment at the center of the metaphase spindle are required to ensure perfect chromosome segregation and to position the cytokinetic furrow that will specify the two daughter cells. Spindle positioning requires regulation of MT dynamics, involving depolymerase activities together with cortical and kinetochore-mediated pushing and pulling forces acting on astral MTs and kinetochore fibres. These forces rely on MT motor activities. Cortical pulling forces exerted on astral MTs depend upon dynein/dynactin complexes and are essential in both symmetric and asymmetric cell division. A well-established spindle positioning pathway regulating the cortical targeting of dynein/dynactin involves the conserved LGN (Leu-Gly-Asn repeat-enriched-protein) and NuMA (microtubule binding nuclear mitotic apparatus protein) complex.1 Spindle orientation is also regulated by integrin-mediated cell adhesion2 and actin retraction fibres that respond to mechanical stress and are influenced by the microenvironment of the dividing cell.3 Altering the capture of astral MTs or modulating pulling forces affects spindle position, which can impair cell division, differentiation and embryogenesis. In this general scheme, the activity of mitotic kinases such as Auroras and Plk1 (Polo-like kinase 1) is crucial.4 Recently, the p21-activated kinases (PAKs) emerged as novel important players in mitotic progression. In our recent article, we demonstrated that PAK4 regulates spindle positioning in symmetric cell division.5 In this commentary, and in light of recent published studies, we discuss how PAK4 could participate in the regulation of mechanisms involved in spindle positioning and orientation. PMID:22960742

  7. Study of Dynamic Features of Surface Plasma in High-Power Disk Laser Welding

    NASA Astrophysics Data System (ADS)

    Wang, Teng; Gao, Xiangdong; Katayama, Seiji; Jin, Xiaoli

    2012-03-01

    High-speed photography was used to obtain the dynamic changes in the surface plasma during a high-power disk laser welding process. A color space clustering algorithm to extract the edge information of the surface plasma region was developed in order to improve the accuracy of image processing. With a comparative analysis of the plasma features, i.e., area and height, and the characteristics of the welded seam, the relationship between the surface plasma and the stability of the laser welding process was characterized, which provides a basic understanding for the real-time monitoring of laser welding.

  8. Mast, a conserved microtubule-associated protein required for bipolar mitotic spindle organization

    PubMed Central

    Lemos, Catarina L.; Sampaio, Paula; Maiato, Helder; Costa, Madalena; Omel’yanchuk, Leonid V.; Liberal, Vasco; Sunkel, Claudio E.

    2000-01-01

    Through mutational analysis in Drosophila, we have identified the gene multiple asters (mast), which encodes a new 165 kDa protein. mast mutant neuroblasts are highly polyploid and show severe mitotic abnormalities including the formation of mono- and multi-polar spindles organized by an irregular number of microtubule-organizing centres of abnormal size and shape. The mast gene product is evolutionarily conserved since homologues were identified from yeast to man, revealing a novel protein family. Antibodies against Mast and analysis of tissue culture cells expressing an enhanced green fluorescent protein–Mast fusion protein show that during mitosis, this protein localizes to centrosomes, the mitotic spindle, centromeres and spindle midzone. Microtubule-binding assays indicate that Mast is a microtubule-associated protein displaying strong affinity for polymerized microtubules. The defects observed in the mutant alleles and the intracellular localization of the protein suggest that Mast plays an essential role in centrosome separation and organization of the bipolar mitotic spindle. PMID:10899121

  9. Mast, a conserved microtubule-associated protein required for bipolar mitotic spindle organization.

    PubMed

    Lemos, C L; Sampaio, P; Maiato, H; Costa, M; Omel'yanchuk, L V; Liberal, V; Sunkel, C E

    2000-07-17

    Through mutational analysis in Drosopjila we have identified the gene multiple asters (mast), which encodes a new 165 kDa protein. mast mutant neuroblasts are highly polyploid and show severe mitotic abnormalities including the formation of mono- and multi-polar spindles organized by an irregular number of microtubule-organizing centres of abnormal size and shape. The mast gene product is evolutionarily conserved since homologues were identified from yeast to man, revealing a novel protein family. Antibodies against Mast and analysis of tissue culture cells expressing an enhanced green fluorescent protein-Mast fusion protein show that during mitosis, this protein localizes to centrosomes, the mitotic spindle, centromeres and spindle midzone. Microtubule-binding assays indicate that Mast is a microtubule-associated protein displaying strong affinity for polymerized microtubules. The defects observed in the mutant alleles and the intracellular localization of the protein suggest that Mast plays an essential role in centrosome separation and organization of the bipolar mitotic spindle. PMID:10899121

  10. Insulin growth factors regulate the mitotic cycle in cultured rat sympathetic neuroblasts

    SciTech Connect

    DiCicco-Bloom, E.; Black, I.B. )

    1988-06-01

    While neuronal mitosis is uniquely restricted to early development, the underlying regulation remains to be defined. The authors have now developed a dissociated, embryonic sympathetic neuron culture system that uses fully defined medium in which cells enter the mitotic cycle. The cultured cells expressed two neuronal traits, tyrosine hydroxylase and the neuron-specific 160-kDa neurofilament subunit protein, but were devoid of glial fibrillary acidic protein, a marker for non-myelin-forming Schwann cells in ganglia. Approximately one-third of the tyrosine hydroxylase-positive cells synthesized DNA in culture, specifically incorporating ({sup 3}H)thymidine into their nuclei. They used this system to define factors regulating the mitotic cycle in sympathetic neuroblasts. Members of the insulin family of growth factors, including insulin and insulin-like growth factors I and II, regulated DNA synthesis in the presumptive neuroblasts. Insulin more than doubled the proportion of tyrosine hydroxylase-positive cells entering the mitotic cycle, as indicated by autoradiography of ({sup 3}H)thymidine incorporation into nuclei. Scintillation spectrometry was an even more sensitive index of DNA synthesis. In contrast, the trophic protein nerve growth factor exhibited no mitogenic effect, suggesting that the mitogenic action of insulin growth factors is highly specific. The observations are discussed in the context of the detection of insulin growth factors and receptors in the developing brain.

  11. Pharmacological inactivation of CHK1 and WEE1 induces mitotic catastrophe in nasopharyngeal carcinoma cells.

    PubMed

    Mak, Joyce P Y; Man, Wing Yu; Chow, Jeremy P H; Ma, Hoi Tang; Poon, Randy Y C

    2015-08-28

    Nasopharyngeal carcinoma (NPC) is a rare but highly invasive cancer. As radiotherapy is the primary treatment for NPC, this offers a rationale to investigate if uncoupling the DNA damage responses can sensitize this cancer type. The G2 DNA damage checkpoint is controlled by a cascade of protein kinases: ATM/ATR, which phosphorylates CHK1/CHK2, which in turn phosphorylates WEE1. A number of small molecule inhibitors have been developed against these kinases as potential therapeutic agents. Here we demonstrated that compare to that in immortalized nasopharyngeal epithelial cells, ATR, CHK1, and WEE1 were overexpressed in NPC cell lines. Inhibitors of these kinases were unable to promote extensive mitotic catastrophe in ionizing radiation-treated NPC cells, indicating that they are not very effective radiosensitizer for this cancer. In the absence of prior irradiation, however, mitotic catastrophe could be induced with inhibitors against CHK1 (AZD7762) or WEE1 (MK-1775). NPC cells were more sensitive to WEE1 inactivation than nasopharyngeal epithelial cells. Targeting CHK1 and WEE1 together induced more extensive mitotic catastrophe than the individual components alone. Taken together, our results show that NPC cells depend on CHK1 and WEE1 activity for growth and that inhibitors of these kinases may serve as potential therapeutics for NPC. PMID:26025928

  12. Pharmacological inactivation of CHK1 and WEE1 induces mitotic catastrophe in nasopharyngeal carcinoma cells

    PubMed Central

    Mak, Joyce P.Y.; Man, Wing Yu; Chow, Jeremy P.H.; Ma, Hoi Tang; Poon, Randy Y.C.

    2015-01-01

    Nasopharyngeal carcinoma (NPC) is a rare but highly invasive cancer. As radiotherapy is the primary treatment for NPC, this offers a rationale to investigate if uncoupling the DNA damage responses can sensitize this cancer type. The G2 DNA damage checkpoint is controlled by a cascade of protein kinases: ATM/ATR, which phosphorylates CHK1/CHK2, which in turn phosphorylates WEE1. A number of small molecule inhibitors have been developed against these kinases as potential therapeutic agents. Here we demonstrated that compare to that in immortalized nasopharyngeal epithelial cells, ATR, CHK1, and WEE1 were overexpressed in NPC cell lines. Inhibitors of these kinases were unable to promote extensive mitotic catastrophe in ionizing radiation-treated NPC cells, indicating that they are not very effective radiosensitizer for this cancer. In the absence of prior irradiation, however, mitotic catastrophe could be induced with inhibitors against CHK1 (AZD7762) or WEE1 (MK-1775). NPC cells were more sensitive to WEE1 inactivation than nasopharyngeal epithelial cells. Targeting CHK1 and WEE1 together induced more extensive mitotic catastrophe than the individual components alone. Taken together, our results show that NPC cells depend on CHK1 and WEE1 activity for growth and that inhibitors of these kinases may serve as potential therapeutics for NPC. PMID:26025928

  13. Twinning and mitotic crossing-over: some possibilities and their implications.

    PubMed Central

    Côté, G B; Gyftodimou, J

    1991-01-01

    Mitotic crossing-over does occur in man and is much more frequent and important than generally assumed. Its postzygotic occurrence before an embryo differentiates into MZ twins is theoretically predicted to have disrupting effects on genomic imprinting and cis-acting sequences, with consequences ranging from early lethality to MZ twin discordance. Some predictions are at odds with classical views on twinning and include a high discordance rate of MZ twins for some genetic diseases. A review of MZ twin discordance and an attempt at explaining some of the data lead one to hypothesize both the existence of a sex differences in the rate of mitotic crossing-over and the impossibility for crossed X chromosomes to undergo inactivation. The close interrelationship of twinning and midline malformations further suggests a major role of mitotic crossing-over in the induction of the twinning process itself. The model can be tested with molecular methods and provides a new approach for the gene mapping of so-called multifactorial diseases and of rarer disorders with apparently irregular inheritance. PMID:2063864

  14. Cytoplasmic flows as signatures for the mechanics of mitotic spindle positioning

    NASA Astrophysics Data System (ADS)

    Nazockdast, Ehssan; Rahimian, Abtin; Needleman, Daniel; Shelley, Michael

    2015-11-01

    The proper positioning of the mitotic spindle is crucial for asymmetric cell division and generating cell diversity during development. We use dynamic simulations to study the cytoplasmic flows generated by three possible active forcing mechanisms involved in positioning of the mitotic spindle in the first cell division of C.elegans embryo namely cortical pulling, cortical pushing, and cytoplasmic pulling mechanisms. The numerical platform we have developed for simulating cytoskeletal assemblies is the first to incorporate the interactions between the fibers and other intracellular bodies with the cytoplasmic fluid, while also accounting for their polymerization, and interactions with motor proteins. The hydrodynamic interactions are computed using boundary integral methods in Stokes flow coupled with highly efficient fast summation techniques that reduce the computational cost to scale linearly with the number of fibers and other bodies. We show that although all three force transduction mechanisms predict proper positioning and orientation of the mitotic spindle, each model produces a different signature in its induced cytoplasmic flow and MT conformation. We suggest that cytoplasmic flows and MT conformation can be used to differentiate between these mechanisms.

  15. Transcriptional response to stress in the dynamic chromatin environment of cycling and mitotic cells

    PubMed Central

    Vihervaara, Anniina; Sergelius, Christian; Vasara, Jenni; Blom, Malin A. H.; Elsing, Alexandra N.; Roos-Mattjus, Pia; Sistonen, Lea

    2013-01-01

    Heat shock factors (HSFs) are the master regulators of transcription under protein-damaging conditions, acting in an environment where the overall transcription is silenced. We determined the genomewide transcriptional program that is rapidly provoked by HSF1 and HSF2 under acute stress in human cells. Our results revealed the molecular mechanisms that maintain cellular homeostasis, including HSF1-driven induction of polyubiquitin genes, as well as HSF1- and HSF2-mediated expression patterns of cochaperones, transcriptional regulators, and signaling molecules. We characterized the genomewide transcriptional response to stress also in mitotic cells where the chromatin is tightly compacted. We found a radically limited binding and transactivating capacity of HSF1, leaving mitotic cells highly susceptible to proteotoxicity. In contrast, HSF2 occupied hundreds of loci in the mitotic cells and localized to the condensed chromatin also in meiosis. These results highlight the importance of the cell cycle phase in transcriptional responses and identify the specific mechanisms for HSF1 and HSF2 in transcriptional orchestration. Moreover, we propose that HSF2 is an epigenetic regulator directing transcription throughout cell cycle progression. PMID:23959860

  16. High-resolution digital elevation models of the Flade Iceblink feature in NE Greenland

    NASA Astrophysics Data System (ADS)

    Willis, M. J.; Juntunen, T.; Porter, C. C.; Morin, P. J.

    2013-12-01

    We produce a time series of high-resolution digital elevation models (DEM) to examine the recent evolution of an 8.7 km2 sub-glacial lake collapse feature near the southern summit of the 8500 km2 Flade Isblink Ice Cap (FIIC) in northeastern Greenland [Figure 1]. Visible imagery from the MODerate-resolution Imaging Spectroradiometer (MODIS) indicates the collapse occurred between August 16th and September 6th, 2011 at the site of a recurring moulin. DEMs are extracted using the NASA Ames Stereo Pipeline for the period between June 2012 and late 2013 from 0.5 m resolution along-track stereo image pairs available via the NGA commercial imagery program. The DEMs are compared to a 1996 ERS InSAR derived DEM [Palmer et al., 2010], and to a contemporary airborne laser altimeter swath flown by NASA Icebridge in mid-April 2013 to derive the volume of the feature and the uncertainties on the high-resolution DEMs. The 'mitten' shaped feature is bounded by crevasses on three sides, with a shallow ramp to the south. It is ~70 m deep, 3.7 km north-to-south and 3 km east-to-west and has a volume of ~0.3 km3. Ice penetrating radar from a nearby Icebridge mission in May 2011, indicates the ice is approximately 550 m thick and that the bed is very flat and smooth about 1 km to the southeast of the feature. The nearby bed topography, local geology and lack of recorded seismicity in the area indicate it is unlikely that the feature is the result of either subglacial volcanic activity or the collapse of a limestone karst feature below the ice cap - the neighboring Princess Elizabeth Alps are composed of 420 Ma Caledonide fold belt gneisses. The presence of recurring supraglacial meltwater streams and drainage into the feature, its rapid formation and its steep sided nature instead suggest that it formed during the rapid drainage of a sub-glacial lake - which is, as far as we are aware, the first recorded instance of such an occurrence in Greenland. Meltwater observed using 250 m resolution MODIS imagery during the extensive melt seasons of both 2010 and 2011 flows northwards into the area of the feature before disappearing - presumably down a Moulin. We use RACMO2 to provide rough estimates of the volumes involved. We monitor the elevation of the floor of the feature to see if the subglacial lake is refilling and provide gross, low-resolution estimates of hydraulic head and drainage path calculations for the region. Flade Iceblink feature from IceBridge. Michael Studdinger/NASA mike.willis@cornell.edu Flade Ice Blink as taken by Michael Studdinger/NASA

  17. High-velocity features of calcium and silicon in the spectra of Type Ia supernovae

    NASA Astrophysics Data System (ADS)

    Silverman, Jeffrey M.; Vinkó, József; Marion, G. H.; Wheeler, J. Craig; Barna, Barnabás; Szalai, Tamás; Mulligan, Brian W.; Filippenko, Alexei V.

    2015-08-01

    `High-velocity features' (HVFs) are spectral features in Type Ia supernovae (SNe Ia) that have minima indicating significantly higher (by greater than about 6000 km s-1) velocities than typical `photospheric-velocity features' (PVFs). The PVFs are absorption features with minima indicating typical photospheric (i.e. bulk ejecta) velocities (usually ˜9000-15 000 km s-1 near B-band maximum brightness). In this work, we undertake the most in-depth study of HVFs ever performed. The data set used herein consists of 445 low-resolution optical and near-infrared (NIR) spectra (at epochs up to 5 d past maximum brightness) of 210 low-redshift SNe Ia that follow the `Phillips relation'. A series of Gaussian functions is fit to the data in order to characterize possible HVFs of Ca II H&K, Si II λ6355, and the Ca II NIR triplet. The temporal evolution of the velocities and strengths of the PVFs and HVFs of these three spectral features is investigated, as are possible correlations with other SN Ia observables. We find that while HVFs of Ca II are regularly observed (except in underluminous SNe Ia, where they are never found), HVFs of Si II λ6355 are significantly rarer, and they tend to exist at the earliest epochs and mostly in objects with large photospheric velocities. It is also shown that stronger HVFs of Si II λ6355 are found in objects that lack C II absorption at early times and that have red ultraviolet/optical colours near maximum brightness. These results lead to a self-consistent connection between the presence and strength of HVFs of Si II λ6355 and many other mutually correlated SN Ia observables, including photospheric velocity.

  18. MYC Is a Major Determinant of Mitotic Cell Fate

    PubMed Central

    Topham, Caroline; Tighe, Anthony; Ly, Peter; Bennett, Ailsa; Sloss, Olivia; Nelson, Louisa; Ridgway, Rachel A.; Huels, David; Littler, Samantha; Schandl, Claudia; Sun, Ying; Bechi, Beatrice; Procter, David J.; Sansom, Owen J.; Cleveland, Don W.; Taylor, Stephen S.

    2015-01-01

    Summary Taxol and other antimitotic agents are frontline chemotherapy agents but the mechanisms responsible for patient benefit remain unclear. Following a genome-wide siRNA screen, we identified the oncogenic transcription factor Myc as a taxol sensitizer. Using time-lapse imaging to correlate mitotic behavior with cell fate, we show that Myc sensitizes cells to mitotic blockers and agents that accelerate mitotic progression. Myc achieves this by upregulating a cluster of redundant pro-apoptotic BH3-only proteins and suppressing pro-survival Bcl-xL. Gene expression analysis of breast cancers indicates that taxane responses correlate positively with Myc and negatively with Bcl-xL. Accordingly, pharmacological inhibition of Bcl-xL restores apoptosis in Myc-deficient cells. These results open up opportunities for biomarkers and combination therapies that could enhance traditional and second-generation antimitotic agents. PMID:26175417

  19. Mitotic remodeling of the replicon and chromosome structure.

    PubMed

    Lemaitre, Jean-Marc; Danis, Etienne; Pasero, Philippe; Vassetzky, Yegor; Méchali, Marcel

    2005-12-01

    Animal cloning by nuclear-transfer experiments frequently fails due to the inability of transplanted nuclei to support normal embryonic development. We show here that the formation of mitotic chromosomes in the egg context is crucial for adapting differentiated nuclei for early development. Differentiated erythrocyte nuclei replicate inefficiently in Xenopus eggs but do so as rapidly as sperm nuclei if a prior single mitosis is permitted. This mitotic remodeling involves a topoisomerase II-dependent shortening of chromatin loop domains and an increased recruitment of replication initiation factors onto chromatin, leading to a short interorigin spacing characteristic of early developmental stages. It also occurs within each early embryonic cell cycle and dominantly regulates initiation of DNA replication for the subsequent S phase. These results indicate that mitotic conditioning is crucial to reset the chromatin structure of differentiated adult donor cells for embryonic DNA replication and suggest that it is an important step in nuclear cloning. PMID:16325575

  20. Mechanisms and Regulation of Mitotic Recombination in Saccharomyces cerevisiae

    PubMed Central

    Symington, Lorraine S.; Rothstein, Rodney; Lisby, Michael

    2014-01-01

    Homology-dependent exchange of genetic information between DNA molecules has a profound impact on the maintenance of genome integrity by facilitating error-free DNA repair, replication, and chromosome segregation during cell division as well as programmed cell developmental events. This chapter will focus on homologous mitotic recombination in budding yeast Saccharomyces cerevisiae. However, there is an important link between mitotic and meiotic recombination (covered in the forthcoming chapter by Hunter et al. 2015) and many of the functions are evolutionarily conserved. Here we will discuss several models that have been proposed to explain the mechanism of mitotic recombination, the genes and proteins involved in various pathways, the genetic and physical assays used to discover and study these genes, and the roles of many of these proteins inside the cell. PMID:25381364

  1. A requirement for epsin in mitotic membrane and spindle organization

    PubMed Central

    2009-01-01

    Eukaryotic cells possess a sophisticated membrane system to facilitate diverse functions. Whereas much is known about the nature of membrane systems in interphase, the organization and function of the mitotic membrane system are less well understood. In this study, we show that epsin, an endocytic adapter protein, regulates mitotic membrane morphology and spindle integrity in HeLa cells. Using epsin that harbors point mutations in the epsin NH2-terminal homology domain and spindle assembly assays in Xenopus laevis egg extracts, we show that epsin-induced membrane curvature is required for proper spindle morphogenesis, independent of its function in endocytosis during interphase. Although several other membrane-interacting proteins, including clathrin, AP2, autosomal recessive hypercholesterolemia, and GRASP65, are implicated in the regulation of mitosis, whether they participate through regulation of membrane organization is unclear. Our study of epsin provides evidence that mitotic membrane organization influences spindle integrity. PMID:19704019

  2. Feature extraction and classification of clouds in high resolution panchromatic satellite imagery

    NASA Astrophysics Data System (ADS)

    Sharghi, Elan

    The development of sophisticated remote sensing sensors is rapidly increasing, and the vast amount of satellite imagery collected is too much to be analyzed manually by a human image analyst. It has become necessary for a tool to be developed to automate the job of an image analyst. This tool would need to intelligently detect and classify objects of interest through computer vision algorithms. Existing software called the Rapid Image Exploitation Resource (RAPIER®) was designed by engineers at Space and Naval Warfare Systems Center Pacific (SSC PAC) to perform exactly this function. This software automatically searches for anomalies in the ocean and reports the detections as a possible ship object. However, if the image contains a high percentage of cloud coverage, a high number of false positives are triggered by the clouds. The focus of this thesis is to explore various feature extraction and classification methods to accurately distinguish clouds from ship objects. An examination of a texture analysis method, line detection using the Hough transform, and edge detection using wavelets are explored as possible feature extraction methods. The features are then supplied to a K-Nearest Neighbors (KNN) or Support Vector Machine (SVM) classifier. Parameter options for these classifiers are explored and the optimal parameters are determined.

  3. Defective in mitotic arrest 1/ring finger 8 is a checkpoint protein that antagonizes the human mitotic exit network.

    PubMed

    Tuttle, Robyn L; Bothos, John; Summers, Matthew K; Luca, Francis C; Halazonetis, Thanos D

    2007-12-01

    A molecular pathway homologous to the S. cerevisiae mitotic exit network (MEN) and S. pombe septation initiation network has recently been described in higher eukaryotes and involves the tumor suppressor kinase LATS1 and its subunit MOB1A. The yeast MEN/septation initiation network pathways are regulated by the ubiquitin ligase defective in mitotic arrest 1 (Dma1p), a checkpoint protein that helps maintain prometaphase arrest when cells are exposed to microtubule poisons. We identified here the RING domain protein ring finger 8 (RNF8) as the human orthologue of the yeast protein Dma1p. Like its yeast counterparts, human DMA1/RNF8 localized at the midbody and its depletion by siRNA compromised mitotic arrest of nocodazole-treated cells in a manner dependent on the MEN. Depletion of MAD2, a spindle checkpoint protein, also compromised mitotic arrest, but in a MEN-independent manner. Thus, two distinct checkpoint pathways maintain mitotic arrest in cells exposed to microtubule poisons. PMID:18171988

  4. Spectral feature design for data compression in high dimensional multispectral data

    NASA Technical Reports Server (NTRS)

    Chen, C.-C. Thomas; Landgrebe, David A.

    1987-01-01

    Data transmission loads of high dimensional remote sensor systems can be greatly reduced by applying generalized Karhunen-Loeve transform as a feature design technique. Two spectral feature design approaches based upon the generalized K-L transform are developed to compress information effectively. Six sets of field data from Kansas and North Dakota on three different dates each are used to test the methods. Spatially, temporally and spatially/temporally combined data sets are formed in this paper to test the robustness property of the schemes. The probability of correct classification using Landsat MSS, Thematic Mapper bands and the proposed bands are found and compared. The comparison shows that the results are improved by the proposed methods, and they appear to be satisfactorily robust. The overall data compression ratio in this paper is about 100/16, i.e., about 6 to 1 with no loss in classification accuracy.

  5. Rohitukine inhibits in vitro adipogenesis arresting mitotic clonal expansion and improves dyslipidemia in vivo[S

    PubMed Central

    Varshney, Salil; Shankar, Kripa; Beg, Muheeb; Balaramnavar, Vishal M.; Mishra, Sunil Kumar; Jagdale, Pankaj; Srivastava, Shishir; Chhonker, Yashpal S.; Lakshmi, Vijai; Chaudhari, Bhushan P.; Bhatta, Rabi Shankar; Saxena, Anil Kumar; Gaikwad, Anil Nilkanth

    2014-01-01

    We developed a common feature pharmacophore model using known antiadipogenic compounds (CFPMA). We identified rohitukine, a reported chromone anticancer alkaloid as a potential hit through in silico mapping of the in-house natural product library on CFPMA. Studies were designed to assess the antiadipogenic potential of rohitukine. Rohitukine was isolated from Dysoxylum binacteriferum Hook. to ⬧95% purity. As predicted by CFPMA, rohitukine was indeed found to be an antiadipogenic molecule. Rohitukine inhibited lipid accumulation and adipogenic differentiation in a concentration- and exposure-time-dependent manner in 3T3-L1 and C3H10T1/2 cells. Rohitukine downregulated expression of PPARγ, CCAAT/enhancer binding protein α, adipocyte protein 2 (aP2), FAS, and glucose transporter 4. It also suppressed mRNA expression of LPL, sterol-regulatory element binding protein (SREBP) 1c, FAS, and aP2, the downstream targets of PPARγ. Rohitukine arrests cells in S phase during mitotic clonal expansion. Rohitukine was bioavailable, and 25.7% of orally administered compound reached systemic circulation. We evaluated the effect of rohitukine on dyslipidemia induced by high-fat diet in the hamster model. Rohitukine increased hepatic expression of liver X receptor α and decreased expression of SREBP-2 and associated targets. Rohitukine decreased hepatic and gonadal lipid accumulation and ameliorated dyslipidemia significantly. In summary, our strategy to identify a novel antiadipogenic molecule using CFPMA successfully resulted in identification of rohitukine, which confirmed antiadipogenic activity and also exhibited in vivo antidyslipidemic activity. PMID:24646949

  6. Single-walled carbon nanotube-induced mitotic disruption⋆

    PubMed Central

    Sargent, L.M.; Hubbs, A.F.; Young, S.-H.; Kashon, M.L.; Dinu, C.Z.; Salisbury, J.L.; Benkovic, S.A.; Lowry, D.T.; Murray, A.R.; Kisin, E.R.; Siegrist, K.J.; Battelli, L.; Mastovich, J.; Sturgeon, J.L.; Bunker, K.L.; Shvedova, A.A.; Reynolds, S.H.

    2015-01-01

    Carbon nanotubes were among the earliest products of nanotechnology and have many potential applications in medicine, electronics, and manufacturing. The low density, small size, and biological persistence of carbon nanotubes create challenges for exposure control and monitoring and make respiratory exposures to workers likely. We have previously shown mitotic spindle aberrations in cultured primary and immortalized human airway epithelial cells exposed to 24, 48 and 96 μg/cm2 single-walled carbon nanotubes (SWCNT). To investigate mitotic spindle aberrations at concentrations anticipated in exposed workers, primary and immortalized human airway epithelial cells were exposed to SWCNT for 24–72 h at doses equivalent to 20 weeks of exposure at the Permissible Exposure Limit for particulates not otherwise regulated. We have now demonstrated fragmented centrosomes, disrupted mitotic spindles and aneuploid chromosome number at those doses. The data further demonstrated multipolar mitotic spindles comprised 95% of the disrupted mitoses. The increased multipolar mitotic spindles were associated with an increased number of cells in the G2 phase of mitosis, indicating a mitotic checkpoint response. Nanotubes were observed in association with mitotic spindle microtubules, the centrosomes and condensed chromatin in cells exposed to 0.024, 0.24, 2.4 and 24 μg/cm2 SWCNT. Three-dimensional reconstructions showed carbon nanotubes within the centrosome structure. The lower doses did not cause cytotoxicity or reduction in colony formation after 24 h; however, after three days, significant cytotoxicity was observed in the SWCNT-exposed cells. Colony formation assays showed an increased proliferation seven days after exposure. Our results show significant disruption of the mitotic spindle by SWCNT at occupationally relevant doses. The increased proliferation that was observed in carbon nanotube-exposed cells indicates a greater potential to pass the genetic damage to daughter cells. Disruption of the centrosome is common in many solid tumors including lung cancer. The resulting aneuploidy is an early event in the progression of many cancers, suggesting that it may play a role in both tumorigenesis and tumor progression. These results suggest caution should be used in the handling and processing of carbon nanotubes. PMID:22178868

  7. Force and the spindle: Mechanical cues in mitotic spindle orientation

    PubMed Central

    Nestor-Bergmann, Alexander; Goddard, Georgina; Woolner, Sarah

    2014-01-01

    The mechanical environment of a cell has a profound effect on its behaviour, from dictating cell shape to driving the transcription of specific genes. Recent studies have demonstrated that mechanical forces play a key role in orienting the mitotic spindle, and therefore cell division, in both single cells and tissues. Whilst the molecular machinery that mediates the link between external force and the mitotic spindle remains largely unknown, it is becoming increasingly clear that this is a widely used mechanism which could prove vital for coordinating cell division orientation across tissues in a variety of contexts. PMID:25080021

  8. Automated Identification of River Hydromorphological Features Using UAV High Resolution Aerial Imagery.

    PubMed

    Casado, Monica Rivas; Gonzalez, Rocio Ballesteros; Kriechbaumer, Thomas; Veal, Amanda

    2015-01-01

    European legislation is driving the development of methods for river ecosystem protection in light of concerns over water quality and ecology. Key to their success is the accurate and rapid characterisation of physical features (i.e., hydromorphology) along the river. Image pattern recognition techniques have been successfully used for this purpose. The reliability of the methodology depends on both the quality of the aerial imagery and the pattern recognition technique used. Recent studies have proved the potential of Unmanned Aerial Vehicles (UAVs) to increase the quality of the imagery by capturing high resolution photography. Similarly, Artificial Neural Networks (ANN) have been shown to be a high precision tool for automated recognition of environmental patterns. This paper presents a UAV based framework for the identification of hydromorphological features from high resolution RGB aerial imagery using a novel classification technique based on ANNs. The framework is developed for a 1.4 km river reach along the river Dee in Wales, United Kingdom. For this purpose, a Falcon 8 octocopter was used to gather 2.5 cm resolution imagery. The results show that the accuracy of the framework is above 81%, performing particularly well at recognising vegetation. These results leverage the use of UAVs for environmental policy implementation and demonstrate the potential of ANNs and RGB imagery for high precision river monitoring and river management. PMID:26556355

  9. High-speed object matching and localization using gradient orientation features

    NASA Astrophysics Data System (ADS)

    Xu, Xinyu; van Beek, Peter; Feng, Xiaofan

    2013-12-01

    In many robotics and automation applications, it is often required to detect a given object and determine its pose (position and orientation) from input images with high speed, high robustness to photometric changes, and high pose accuracy. We propose a new object matching method that improves efficiency over existing approaches by decomposing orientation and position estimation into two cascade steps. In the first step, an initial position and orientation is found by matching with Histogram of Oriented Gradients (HOG), reducing orientation search from 2D template matching to 1D correlation matching. In the second step, a more precise orientation and position is computed by matching based on Dominant Orientation Template (DOT), using robust edge orientation features. The cascade combination of the HOG and DOT feature for high-speed and robust object matching is the key novelty of the proposed method. Experimental evaluation was performed with real-world single-object and multi-object inspection datasets, using software implementations on an Atom CPU platform. Our results show that the proposed method achieves significant speed improvement compared to an already accelerated template matching method at comparable accuracy performance.

  10. Automated Identification of River Hydromorphological Features Using UAV High Resolution Aerial Imagery

    PubMed Central

    Rivas Casado, Monica; Ballesteros Gonzalez, Rocio; Kriechbaumer, Thomas; Veal, Amanda

    2015-01-01

    European legislation is driving the development of methods for river ecosystem protection in light of concerns over water quality and ecology. Key to their success is the accurate and rapid characterisation of physical features (i.e., hydromorphology) along the river. Image pattern recognition techniques have been successfully used for this purpose. The reliability of the methodology depends on both the quality of the aerial imagery and the pattern recognition technique used. Recent studies have proved the potential of Unmanned Aerial Vehicles (UAVs) to increase the quality of the imagery by capturing high resolution photography. Similarly, Artificial Neural Networks (ANN) have been shown to be a high precision tool for automated recognition of environmental patterns. This paper presents a UAV based framework for the identification of hydromorphological features from high resolution RGB aerial imagery using a novel classification technique based on ANNs. The framework is developed for a 1.4 km river reach along the river Dee in Wales, United Kingdom. For this purpose, a Falcon 8 octocopter was used to gather 2.5 cm resolution imagery. The results show that the accuracy of the framework is above 81%, performing particularly well at recognising vegetation. These results leverage the use of UAVs for environmental policy implementation and demonstrate the potential of ANNs and RGB imagery for high precision river monitoring and river management. PMID:26556355

  11. Statistical features of the high-latitude ionospheric convection structure associated with enhanced solar wind fluctuations

    NASA Astrophysics Data System (ADS)

    Kim, H.; Lyons, L. R.; Ruohoniemi, J. M.; Frissell, N. A.

    2012-12-01

    While the IMF and solar wind dynamic pressure almost certainly play larger roles under most conditions, evidence has been recently found that Ultra Low Frequency (ULF) wave power in the solar wind has an additional substantial effect on the strength of convection within the polar caps, and on the nightside within both the aurora ionosphere and the plasma sheet. An initial study shows that the convection flows under enhanced solar wind fluctuations often appear to be more structured, with localized strong vortical features, than the convection under steady solar wind conditions. In this work, we statistically examine characteristic features of the ionospheric convection structure in terms of vortex patterns and how they are related to the convection enhancements during periods of enhanced solar wind fluctuations. Specifically, we examine whether enhanced solar wind ULF power will drive localized turbulence within enhanced convection cells while it increases convection strength at the same time. The results of this study will provide evidence for how solar wind ULF fluctuations can contribute to the solar wind energy transfer to the magnetosphere-ionosphere system. To determine the features of 2-D convection structure, we analyze the large-scale global convection maps derived from the SuperDARN observations with extensive radar echo coverage over a large portion of the high latitude ionosphere. Wind and ACE data are used for examination of solar wind and IMF conditions.

  12. High-Velocity Features in the Spectra of Type-Ia Supernovae

    NASA Astrophysics Data System (ADS)

    Silverman, Jeffrey M.; Marion, Howie; Vinko, Jozsef; Mulligan, Brian W.; Wheeler, J. Craig; Filippenko, Alexei V.

    2015-01-01

    Spectra of Type-Ia supernovae (SNe Ia) obtained before maximum brightness sometimes show high-velocity features (HVFs). They are most often seen in Si II and Ca II and in the most obvious cases appear as a second, separate absorption feature at ~7000-10000 km/s higher expansion velocity than the more normal photospheric-velocity features (PVFs). We have investigated how often HVFs occur, at what epochs, and how they evolve with time using a large sample of low-resolution, optical and NIR spectra of nearby SNe Ia. Our ongoing study indicates that HVFs are quite common in SNe Ia spectra obtained prior to 5 days before maximum brightness. Correlations between photometric observables and the relative line strengths and expansion velocities of both HVFs and PVFs are currently being sought and some intriguing results have already been found and will be discussed. Various explanations for the existence and behavior of the HVFs are being considered, with possibilities including density enhancements in the outer portion of the SN ejecta and low levels of interaction with circumstellar material.

  13. Plasma etching of high-resolution features in a fullerene molecular resist

    NASA Astrophysics Data System (ADS)

    Manyam, J.; Manickam, M.; Preece, J. A.; Palmer, R. E.; Robinson, A. P. G.

    2011-04-01

    As resist films become thinner, so as to reduce problems of aspect ratio related pattern collapse at high-resolution, it is becoming increasingly difficult to transfer patterns with useful aspect ratio by directly etching the resist. It has become common to use the photoresist to pattern an intermediate hardmask, which then protects the silicon substrate during etching, allowing useful aspect ratios but adding process complexity. We have previously described a fullerene based electron beam lithography resist capable of 20 nm halfpitch and 12 nm sparse features, at a sensitivity of less than 10 ?C/cm2 at 20 keV. The fullerene resist has high etch durability - comparable to that of commercial novolac resists - and has previously demonstrated an etch selectivity of 3:1 to silicon using electron cyclotron resonance microwave plasma etching with SF6. Here a study of the capabilities of this resist when using Inductively Coupled Plasma etching is presented. Line-space patterns with half-pitches in the range 25 nm to 100 nm, together with sparse features (~20 nm linewidth on a 200 nm pitch) were produced in ~30 nm thick resist films using electron beam lithography, and transferred to silicon using an inductively coupled plasma etcher. Several combinations of SF6, CF4, CHF3 and C4F8process gases were explored. Etch selectivity and anisotropy were studied as a range of etching parameters, such as ICP and RF power, gas flow rate, pressure and temperature were varied. Etch selectivities in excess of 9:1 were demonstrated. Techniques for minimizing aspect ratio dependent etching effects in dense features, including the use of ashing or high etching pressures were also examined.

  14. High-throughput screening for thermoelectric sulphides by using crystal structure features as descriptors

    NASA Astrophysics Data System (ADS)

    Zhang, Ruizhi; Du, Baoli; Chen, Kan; Reece, Mike; Materials Research Insititute Team

    With the increasing computational power and reliable databases, high-throughput screening is playing a more and more important role in the search of new thermoelectric materials. Rather than the well established density functional theory (DFT) calculation based methods, we propose an alternative approach to screen for new TE materials: using crystal structural features as 'descriptors'. We show that a non-distorted transition metal sulphide polyhedral network can be a good descriptor for high power factor according to crystal filed theory. By using Cu/S containing compounds as an example, 1600+ Cu/S containing entries in the Inorganic Crystal Structure Database (ICSD) were screened, and of those 84 phases are identified as promising thermoelectric materials. The screening results are validated by both electronic structure calculations and experimental results from the literature. We also fabricated some new compounds to test our screening results. Another advantage of using crystal structure features as descriptors is that we can easily establish structural relationships between the identified phases. Based on this, two material design approaches are discussed: 1) High-pressure synthesis of metastable phase; 2) In-situ 2-phase composites with coherent interface. This work was supported by a Marie Curie International Incoming Fellowship of the European Community Human Potential Program.

  15. Cyclic development of igneous features and their relationship to high-temperature hydrothermal features in the Henderson porphyry molybdenum deposit, Colorado

    USGS Publications Warehouse

    Carten, R.B.; Geraghty, E.P.; Walker, B.M.

    1988-01-01

    The Henderson porphyry molybdenum deposit was formed by the superposition of coupled alteration and mineralization events, of varying intensity and size, that were associated with each of at least 11 intrusions. Deposition of molybdenite was accompanied by time-equivalent silicic and potassic alteration. High-temperature alteration and mineralization are spatially and temporally linked to the crystallization of compositionally zoned magma in the apex of stocks. Differences in hydrothermal features associated with each intrusion (e.g., mass of ore, orientation and type of veins, density of veins, and intensity of alteration) correlate with differences in primary igneous features (e.g., composition, texture, morphology, and size). The systematic relations between hydrothermal and magmatic features suggest that primary magma compositions, including volatile contents, largely control the geometry, volume, level of emplacement, and mechanisms of crystallization of stocks. These elements in turn govern the orientations and densities of fractures, which ultimately determine the distribution patterns of hydrothermal alteration and mineralization. -from Authors

  16. DESIGN SAFETY FEATURES OF THE BNL HIGH-TEMPERATURE COMBUSTION FACILITY

    SciTech Connect

    GINSBERG,T.; CICCARELLI,G.; BOCCIO,J.

    2000-06-11

    The Brookhaven National Laboratory (BNL) High-Temperature Combustion Facility (HTCF) was used to perform hydrogen deflagration and detonation experiments at temperatures to 650 K. Safety features that were designed to ensure safe and reliable operation of the experimental program are described. Deflagration and detonation experiments have been conducted using mixtures of hydrogen, air, and steam. Detonation cell size measurements were made as a function of mixture composition and thermodynamic gas conditions. Deflagration-to-detonation transition experiments were also conducted. Results of the experimental program are presented, and implications with respect to hydrogen safety are discussed.

  17. The human papillomavirus type 16 E6 and E7 oncoproteins cooperate to induce mitotic defects and genomic instability by uncoupling centrosome duplication from the cell division cycle

    PubMed Central

    Duensing, Stefan; Lee, Lily Y.; Duensing, Anette; Basile, John; Piboonniyom, Siribang-on; Gonzalez, Sonia; Crum, Christopher P.; Münger, Karl

    2000-01-01

    Loss of genomic integrity is a defining feature of many human malignancies, including human papillomavirus (HPV)-associated preinvasive and invasive genital squamous lesions. Here we show that aberrant mitotic spindle pole formation caused by abnormal centrosome numbers represents an important mechanism in accounting for numeric chromosomal alterations in HPV-associated carcinogenesis. Similar to what we found in histopathological specimens, HPV-16 E6 and E7 oncoproteins cooperate to induce abnormal centrosome numbers, aberrant mitotic spindle pole formation, and genomic instability. The low-risk HPV-6 E6 and E7 proteins did not induce such abnormalities. Whereas the HPV-16 E6 oncoprotein has no immediate effects on centrosome numbers, HPV-16 E7 rapidly induces abnormal centrosome duplication. Thus our results suggest a model whereby HPV-16 E7 induces centrosome-related mitotic disturbances that are potentiated by HPV-16 E6. PMID:10944189

  18. Aurora-A-Dependent Control of TACC3 Influences the Rate of Mitotic Spindle Assembly

    PubMed Central

    Joseph, Nimesh; Cavazza, Tommaso; Vernos, Isabelle; Pfuhl, Mark; Gergely, Fanni; Bayliss, Richard

    2015-01-01

    The essential mammalian gene TACC3 is frequently mutated and amplified in cancers and its fusion products exhibit oncogenic activity in glioblastomas. TACC3 functions in mitotic spindle assembly and chromosome segregation. In particular, phosphorylation on S558 by the mitotic kinase, Aurora-A, promotes spindle recruitment of TACC3 and triggers the formation of a complex with ch-TOG-clathrin that crosslinks and stabilises kinetochore microtubules. Here we map the Aurora-A-binding interface in TACC3 and show that TACC3 potently activates Aurora-A through a domain centered on F525. Vertebrate cells carrying homozygous F525A mutation in the endogenous TACC3 loci exhibit defects in TACC3 function, namely perturbed localization, reduced phosphorylation and weakened interaction with clathrin. The most striking feature of the F525A cells however is a marked shortening of mitosis, at least in part due to rapid spindle assembly. F525A cells do not exhibit chromosome missegregation, indicating that they undergo fast yet apparently faithful mitosis. By contrast, mutating the phosphorylation site S558 to alanine in TACC3 causes aneuploidy without a significant change in mitotic duration. Our work has therefore defined a regulatory role for the Aurora-A-TACC3 interaction beyond the act of phosphorylation at S558. We propose that the regulatory relationship between Aurora-A and TACC3 enables the transition from the microtubule-polymerase activity of TACC3-ch-TOG to the microtubule-crosslinking activity of TACC3-ch-TOG-clathrin complexes as mitosis progresses. Aurora-A-dependent control of TACC3 could determine the balance between these activities, thereby influencing not only spindle length and stability but also the speed of spindle formation with vital consequences for chromosome alignment and segregation. PMID:26134678

  19. Aurora-A-Dependent Control of TACC3 Influences the Rate of Mitotic Spindle Assembly.

    PubMed

    Burgess, Selena G; Peset, Isabel; Joseph, Nimesh; Cavazza, Tommaso; Vernos, Isabelle; Pfuhl, Mark; Gergely, Fanni; Bayliss, Richard

    2015-07-01

    The essential mammalian gene TACC3 is frequently mutated and amplified in cancers and its fusion products exhibit oncogenic activity in glioblastomas. TACC3 functions in mitotic spindle assembly and chromosome segregation. In particular, phosphorylation on S558 by the mitotic kinase, Aurora-A, promotes spindle recruitment of TACC3 and triggers the formation of a complex with ch-TOG-clathrin that crosslinks and stabilises kinetochore microtubules. Here we map the Aurora-A-binding interface in TACC3 and show that TACC3 potently activates Aurora-A through a domain centered on F525. Vertebrate cells carrying homozygous F525A mutation in the endogenous TACC3 loci exhibit defects in TACC3 function, namely perturbed localization, reduced phosphorylation and weakened interaction with clathrin. The most striking feature of the F525A cells however is a marked shortening of mitosis, at least in part due to rapid spindle assembly. F525A cells do not exhibit chromosome missegregation, indicating that they undergo fast yet apparently faithful mitosis. By contrast, mutating the phosphorylation site S558 to alanine in TACC3 causes aneuploidy without a significant change in mitotic duration. Our work has therefore defined a regulatory role for the Aurora-A-TACC3 interaction beyond the act of phosphorylation at S558. We propose that the regulatory relationship between Aurora-A and TACC3 enables the transition from the microtubule-polymerase activity of TACC3-ch-TOG to the microtubule-crosslinking activity of TACC3-ch-TOG-clathrin complexes as mitosis progresses. Aurora-A-dependent control of TACC3 could determine the balance between these activities, thereby influencing not only spindle length and stability but also the speed of spindle formation with vital consequences for chromosome alignment and segregation. PMID:26134678

  20. New insights into the pathology of podocyte loss: mitotic catastrophe.

    PubMed

    Liapis, Helen; Romagnani, Paola; Anders, Hans-Joachim

    2013-11-01

    Podocytes represent an essential component of the kidney's glomerular filtration barrier. They stay attached to the glomerular basement membrane via integrin interactions that support the capillary wall to withstand the pulsating filtration pressure. Podocyte structure is maintained by a dynamic actin cytoskeleton. Terminal differentiation is coupled with permanent exit from the cell cycle and arrest in a postmitotic state. Postmitotic podocytes do not have an infinite life span; in fact, physiologic loss in the urine is documented. Proteinuria and other injuries accelerate podocyte loss or induce death. Mature podocytes are unable to replicate and maintain their actin cytoskeleton simultaneously. By the end of mitosis, cytoskeletal actin forms part of the contractile ring, rendering a round shape to podocytes. Therefore, when podocyte mitosis is attempted, it may lead to aberrant mitosis (ie, mitotic catastrophe). Mitotic catastrophe implies that mitotic podocytes eventually detach or die; this is a previously unrecognized form of podocyte loss and a compensatory mechanism for podocyte hypertrophy that relies on post-G1-phase cell cycle arrest. In contrast, local podocyte progenitors (parietal epithelial cells) exhibit a simple actin cytoskeleton structure and can easily undergo mitosis, supporting podocyte regeneration. In this review we provide an appraisal of the in situ pathology of mitotic catastrophe compared with other proposed types of podocyte death and put experimental and renal biopsy data in a unified perspective. PMID:24007883

  1. O-GlcNAc Transferase Regulates Mitotic Chromatin Dynamics*

    PubMed Central

    Sakabe, Kaoru; Hart, Gerald W.

    2010-01-01

    Mitosis must faithfully divide the genome such that each progeny inherits the same genetic material. DNA condensation is crucial in ensuring that chromosomes are correctly attached to the mitotic spindle for segregation, preventing DNA breaks or constrictions from the contractile ring. Histones form an octameric complex of basic proteins important in regulating DNA organization and accessibility. Histone post-translational modifications are altered during mitosis, although the roles of these post-translational modifications remain poorly characterized. Here, we report that N-acetylglucosamine (O-GlcNAc) transferase (OGT), the enzyme catalyzing the addition of O-GlcNAc moieties to nuclear and cytoplasmic proteins at serine and threonine residues, regulates some aspects of mitotic chromatin dynamics. OGT protein amounts decrease during M phase. Modest overexpression of OGT alters mitotic histone post-translational modifications at Lys-9, Ser-10, Arg-17, and Lys-27 of histone H3. Overexpression of OGT also prevents mitotic phosphorylation of coactivator-associated arginine methyltransferase 1 (CARM1) and prevents its correct cellular localization during mitosis. Moreover, OGT overexpression results in an increase in abnormal chromosomal bridge formation. Together, these results show that regulating the amount of OGT during mitosis is important in ensuring correct chromosomal segregation during mitosis. PMID:20805223

  2. Rab11 endosomes contribute to mitotic spindle organization and orientation.

    PubMed

    Hehnly, Heidi; Doxsey, Stephen

    2014-03-10

    During interphase, Rab11-GTPase-containing endosomes recycle endocytic cargo. However, little is known about Rab11 endosomes in mitosis. Here, we show that Rab11 localizes to the mitotic spindle and regulates dynein-dependent endosome localization at poles. We found that mitotic recycling endosomes bind γ-TuRC components and associate with tubulin in vitro. Rab11 depletion or dominant-negative Rab11 expression disrupts astral microtubules, delays mitosis, and redistributes spindle pole proteins. Reciprocally, constitutively active Rab11 increases astral microtubules, restores γ-tubulin spindle pole localization, and generates robust spindles. This suggests a role for Rab11 activity in spindle pole maturation during mitosis. Rab11 depletion causes misorientation of the mitotic spindle and the plane of cell division. These findings suggest a molecular mechanism for the organization of astral microtubules and the mitotic spindle through Rab11-dependent control of spindle pole assembly and function. We propose that Rab11 and its associated endosomes cocontribute to these processes through retrograde transport to poles by dynein. PMID:24561039

  3. Occludin Localizes to Centrosomes and Modifies Mitotic Entry*

    PubMed Central

    Runkle, E. Aaron; Sundstrom, Jeffrey M.; Runkle, Kristin B.; Liu, Xuwen; Antonetti, David A.

    2011-01-01

    Proper control of cell cycle progression and barrier function are essential processes to the maintenance of epithelial cell homeostasis. The contribution of tight junction proteins to barrier function is well established, whereas their contribution to cell cycle control is only beginning to be understood. Centrosomes are the principal microtubule organizing centers in eukaryotic cells and centrosome duplication and separation are linked to the cell cycle and mitotic entry. Here we demonstrate that occludin localizes with centrosomes in Madin-Darby canine kidney cells. Immunocytochemistry and biochemical fractionation studies reveal occludin localizes with centrosomes during interphase and occludin Ser-490 phosphorylation at centrosomes increases with mitotic entry. Stable expression of aspartic acid phosphomimetic (S490D) results in centrosomal localization of occludin and increases cell numbers. Furthermore, we provide evidence that occludin regulates centrosome separation and mitotic entry as the nonphosphorylatable alanine mutation (S490A) impedes centrosome separation, delays mitotic entry, and reduces proliferation. Collectively, these studies demonstrate a novel location and function for occludin in centrosome separation and mitosis. PMID:21757728

  4. Mitotic phosphorylation of nucleoporins: dismantling NPCs and beyond.

    PubMed

    Doye, Valérie

    2011-03-15

    Recently reporting in Cell, Laurell et al. (2011) demonstrate that the hyperphosphorylation of vertebrate Nup98 by distinct mitotic kinases contributes to its release from nuclear pores, drives nuclear envelope permeabilization, and may provide a molecular switch coordinating nuclear envelope breakdown and spindle formation. PMID:21397836

  5. Rab11-endosomes contribute to mitotic spindle orientation

    PubMed Central

    Hehnly, Heidi; Doxsey, Stephen

    2014-01-01

    During interphase, Rab11-GTPase-containing endosomes recycle endocytic cargo. However, little is known about Rab11 and endosomes in mitosis. Here we show that Rab11 localizes to the mitotic spindle and regulates dynein-dependent endosome localization at poles. We found that mitotic recycling endosomes bind γ-TuRC components and associate with tubulin in vitro. Rab11-depletion or dominant-negative Rab11 expression disrupts astral microtubules, delays mitosis, and redistributes spindle pole proteins. Reciprocally, constitutively-active Rab11 increases astral microtubules, restores γ-tubulin spindle pole localization and generates robust spindles. This suggests a fundamental role for Rab11 activity in spindle pole maturation during mitosis. Rab11 depletion causes misorientation of the mitotic spindle and the plane of cell division. These findings suggest a molecular mechanism for the organization of astral microtubules and the mitotic spindle through Rab11-dependent control of spindle pole assembly and function. We propose that Rab11 and its associated endosomes co-contribute to these processes through retrograde transport to poles by dynein. PMID:24561039

  6. MITOTIC ABNORMALITIES IN SEA URCHIN EMBRYOS EXPOSED TO DACTINOMYCIN*

    PubMed Central

    Kiefer, Barry I.; Entelis, Charles F.; Infante, Anthony A.

    1969-01-01

    Dactinomycin at concentrations of 10, 20, 25, and 50 μg/ml causes mitotic abnormalities in sea urchin embryos. Sister chromosome separation is impaired and anaphase bridges are frequently formed. The result is an unequal distribution of the chromosome complement to daughter cells. Possible explanations are discussed. Images PMID:4905991

  7. Water Extraction in High Resolution Remote Sensing Image Based on Hierarchical Spectrum and Shape Features

    NASA Astrophysics Data System (ADS)

    Li, Bangyu; Zhang, Hui; Xu, Fanjiang

    2014-03-01

    This paper addresses the problem of water extraction from high resolution remote sensing images (including R, G, B, and NIR channels), which draws considerable attention in recent years. Previous work on water extraction mainly faced two difficulties. 1) It is difficult to obtain accurate position of water boundary because of using low resolution images. 2) Like all other image based object classification problems, the phenomena of "different objects same image" or "different images same object" affects the water extraction. Shadow of elevated objects (e.g. buildings, bridges, towers and trees) scattered in the remote sensing image is a typical noise objects for water extraction. In many cases, it is difficult to discriminate between water and shadow in a remote sensing image, especially in the urban region. We propose a water extraction method with two hierarchies: the statistical feature of spectral characteristic based on image segmentation and the shape feature based on shadow removing. In the first hierarchy, the Statistical Region Merging (SRM) algorithm is adopted for image segmentation. The SRM includes two key steps: one is sorting adjacent regions according to a pre-ascertained sort function, and the other one is merging adjacent regions based on a pre-ascertained merging predicate. The sort step is done one time during the whole processing without considering changes caused by merging which may cause imprecise results. Therefore, we modify the SRM with dynamic sort processing, which conducts sorting step repetitively when there is large adjacent region changes after doing merging. To achieve robust segmentation, we apply the merging region with six features (four remote sensing image bands, Normalized Difference Water Index (NDWI), and Normalized Saturation-value Difference Index (NSVDI)). All these features contribute to segment image into region of object. NDWI and NSVDI are discriminate between water and some shadows. In the second hierarchy, we adopt the shape features to remove more shadows. The water polygons are generated by vectorization algorithm after water segmentation, and then some shape parameters (Compact, Critical Point and Symmetry) are considered to remove shadow. To evaluate the performance of the proposed method, we collect several Quick Bird images at 0.61-m resolution which are acquired in May 2009 at GUANGZHOU province of China. The proposed method is compared with four other methods in water extraction, including pixel-based segmentation by NDWI, Mean-sift segmentation by NDWI, and SVM with different channels. Experimental results show that the proposed method can increase extraction accuracy and reduce the influence of shadows.

  8. Cytotoxic effects of cylindrospermopsin in mitotic and non-mitotic Vicia faba cells.

    PubMed

    Garda, Tamás; Riba, Milán; Vasas, Gábor; Beyer, Dániel; M-Hamvas, Márta; Hajdu, Gréta; Tándor, Ildikó; Máthé, Csaba

    2015-02-01

    Cylindrospermopsin (CYN) is a cyanobacterial toxin known as a eukaryotic protein synthesis inhibitor. We aimed to study its effects on growth, stress responses and mitosis of a eukaryotic model, Vicia faba (broad bean). Growth responses depended on exposure time (3 or 6d), cyanotoxin concentration, culture conditions (dark or continuous light) and V. faba cultivar ("Standard" or "ARC Egypt Cross"). At 6d of exposure, CYN had a transient stimulatory effect on root system growth, roots being possibly capable of detoxification. The toxin induced nucleus fragmentation, blebbing and chromosomal breaks indicating double stranded DNA breaks and programmed cell death. Root necrotic tissue was observed at 0.1-20 μg mL(-1) CYN that probably impeded toxin uptake into vascular tissue. Growth and cell death processes observed were general stress responses. In lateral root tip meristems, lower CYN concentrations (0.01-0.1 μg mL(-1)) induced the stimulation of mitosis and distinct mitotic phases, irrespective of culture conditions or the cultivar used. Higher cyanotoxin concentrations inhibited mitosis. Short-term exposure of hydroxylurea-synchronized roots to 5 μg mL(-1) CYN induced delay of mitosis that might have been related to a delay of de novo protein synthesis. CYN induced the formation of double, split and asymmetric preprophase bands (PPBs), in parallel with the alteration of cell division planes, related to the interference of cyanotoxin with protein synthesis, thus it was a plant- and CYN specific alteration. PMID:25016338

  9. Mitotic quiescence, but not unique "stemness," marks the phenotype of bone metastasis-initiating cells in prostate cancer.

    PubMed

    Wang, Ning; Docherty, Freyja; Brown, Hannah K; Reeves, Kim; Fowles, Anne; Lawson, Michelle; Ottewell, Penelope D; Holen, Ingunn; Croucher, Peter I; Eaton, Colby L

    2015-08-01

    This study aimed to identify subpopulations of prostate cancer cells that are responsible for the initiation of bone metastases. Using rapidly dividing human prostate cancer cell lines, we identified mitotically quiescent subpopulations (<1%), which we compared with the rapidly dividing populations for patterns of gene expression and for their ability to migrate to the skeletons of athymic mice. The study used 2-photon microscopy to map the presence/distribution of fluorescently labeled, quiescent cells and luciferase expression to determine the presence of growing bone metastases. We showed that the mitotically quiescent cells were very significantly more tumorigenic in forming bone metastases than fast-growing cells (55 vs. 15%) and had a unique gene expression profile. The quiescent cells were not uniquely stem cell like, with no expression of CD133 but had the same level expression of other putative prostate stem cell markers (CD44 and integrins α2/β1), when compared to the rapidly proliferating population. In addition, mitotic quiescence was associated with very high levels of C-X-C chemokine receptor type 4 (CXCR4) production. Inhibition of CXCR4 activity altered the homing of quiescent tumor cells to bone. Our studies suggest that mitotic dormancy is a unique phenotype that facilitates tumor cell colonization of the skeleton in prostate cancer. PMID:25888599

  10. Mio depletion links mTOR regulation to Aurora A and Plk1 activation at mitotic centrosomes

    PubMed Central

    Trinkle-Mulcahy, Laura; Porter, Michael; Jeyaprakash, A. Arockia

    2015-01-01

    Coordination of cell growth and proliferation in response to nutrient supply is mediated by mammalian target of rapamycin (mTOR) signaling. In this study, we report that Mio, a highly conserved member of the SEACAT/GATOR2 complex necessary for the activation of mTORC1 kinase, plays a critical role in mitotic spindle formation and subsequent chromosome segregation by regulating the proper concentration of active key mitotic kinases Plk1 and Aurora A at centrosomes and spindle poles. Mio-depleted cells showed reduced activation of Plk1 and Aurora A kinase at spindle poles and an impaired localization of MCAK and HURP, two key regulators of mitotic spindle formation and known substrates of Aurora A kinase, resulting in spindle assembly and cytokinesis defects. Our results indicate that a major function of Mio in mitosis is to regulate the activation/deactivation of Plk1 and Aurora A, possibly by linking them to mTOR signaling in a pathway to promote faithful mitotic progression. PMID:26124292

  11. Mio depletion links mTOR regulation to Aurora A and Plk1 activation at mitotic centrosomes.

    PubMed

    Platani, Melpomeni; Trinkle-Mulcahy, Laura; Porter, Michael; Jeyaprakash, A Arockia; Earnshaw, William C

    2015-07-01

    Coordination of cell growth and proliferation in response to nutrient supply is mediated by mammalian target of rapamycin (mTOR) signaling. In this study, we report that Mio, a highly conserved member of the SEACAT/GATOR2 complex necessary for the activation of mTORC1 kinase, plays a critical role in mitotic spindle formation and subsequent chromosome segregation by regulating the proper concentration of active key mitotic kinases Plk1 and Aurora A at centrosomes and spindle poles. Mio-depleted cells showed reduced activation of Plk1 and Aurora A kinase at spindle poles and an impaired localization of MCAK and HURP, two key regulators of mitotic spindle formation and known substrates of Aurora A kinase, resulting in spindle assembly and cytokinesis defects. Our results indicate that a major function of Mio in mitosis is to regulate the activation/deactivation of Plk1 and Aurora A, possibly by linking them to mTOR signaling in a pathway to promote faithful mitotic progression. PMID:26124292

  12. Polyglutamylated Tubulin Binding Protein C1orf96/CSAP Is Involved in Microtubule Stabilization in Mitotic Spindles

    PubMed Central

    Ohta, Shinya; Hamada, Mayako; Sato, Nobuko; Toramoto, Iyo

    2015-01-01

    The centrosome-associated C1orf96/Centriole, Cilia and Spindle-Associated Protein (CSAP) targets polyglutamylated tubulin in mitotic microtubules (MTs). Loss of CSAP causes critical defects in brain development; however, it is unclear how CSAP association with MTs affects mitosis progression. In this study, we explored the molecular mechanisms of the interaction of CSAP with mitotic spindles. Loss of CSAP caused MT instability in mitotic spindles and resulted in mislocalization of Nuclear protein that associates with the Mitotic Apparatus (NuMA), with defective MT dynamics. Thus, CSAP overload in the spindles caused extensive MT stabilization and recruitment of NuMA. Moreover, MT stabilization by CSAP led to high levels of polyglutamylation on MTs. MT depolymerization by cold or nocodazole treatment was inhibited by CSAP binding. Live-cell imaging analysis suggested that CSAP-dependent MT-stabilization led to centrosome-free MT aster formation immediately upon nuclear envelope breakdown without γ-tubulin. We therefore propose that CSAP associates with MTs around centrosomes to stabilize MTs during mitosis, ensuring proper bipolar spindle formation and maintenance. PMID:26562023

  13. High-precision image aided inertial navigation with known features: observability analysis and performance evaluation.

    PubMed

    Jiang, Weiping; Wang, Li; Niu, Xiaoji; Zhang, Quan; Zhang, Hui; Tang, Min; Hu, Xiangyun

    2014-01-01

    A high-precision image-aided inertial navigation system (INS) is proposed as an alternative to the carrier-phase-based differential Global Navigation Satellite Systems (CDGNSSs) when satellite-based navigation systems are unavailable. In this paper, the image/INS integrated algorithm is modeled by a tightly-coupled iterative extended Kalman filter (IEKF). Tightly-coupled integration ensures that the integrated system is reliable, even if few known feature points (i.e., less than three) are observed in the images. A new global observability analysis of this tightly-coupled integration is presented to guarantee that the system is observable under the necessary conditions. The analysis conclusions were verified by simulations and field tests. The field tests also indicate that high-precision position (centimeter-level) and attitude (half-degree-level)-integrated solutions can be achieved in a global reference. PMID:25330046

  14. Physiological and genomic features of highly alkaliphilic hydrogen-utilizing Betaproteobacteria from a continental serpentinizing site

    PubMed Central

    Suzuki, Shino; Kuenen, J. Gijs; Schipper, Kira; van der Velde, Suzanne; Ishii, Shun’ichi; Wu, Angela; Sorokin, Dimitry Y.; Tenney, Aaron; Meng, XianYing; Morrill, Penny L.; Kamagata, Yoichi; Muyzer, Gerard; Nealson, Kenneth H.

    2014-01-01

    Serpentinization, or the aqueous alteration of ultramafic rocks, results in challenging environments for life in continental sites due to the combination of extremely high pH, low salinity and lack of obvious electron acceptors and carbon sources. Nevertheless, certain Betaproteobacteria have been frequently observed in such environments. Here we describe physiological and genomic features of three related Betaproteobacterial strains isolated from highly alkaline (pH 11.6) serpentinizing springs at The Cedars, California. All three strains are obligate alkaliphiles with an optimum for growth at pH 11 and are capable of autotrophic growth with hydrogen, calcium carbonate and oxygen. The three strains exhibit differences, however, regarding the utilization of organic carbon and electron acceptors. Their global distribution and physiological, genomic and transcriptomic characteristics indicate that the strains are adapted to the alkaline and calcium-rich environments represented by the terrestrial serpentinizing ecosystems. We propose placing these strains in a new genus ‘Serpentinomonas’. PMID:24845058

  15. High-Precision Image Aided Inertial Navigation with Known Features: Observability Analysis and Performance Evaluation

    PubMed Central

    Jiang, Weiping; Wang, Li; Niu, Xiaoji; Zhang, Quan; Zhang, Hui; Tang, Min; Hu, Xiangyun

    2014-01-01

    A high-precision image-aided inertial navigation system (INS) is proposed as an alternative to the carrier-phase-based differential Global Navigation Satellite Systems (CDGNSSs) when satellite-based navigation systems are unavailable. In this paper, the image/INS integrated algorithm is modeled by a tightly-coupled iterative extended Kalman filter (IEKF). Tightly-coupled integration ensures that the integrated system is reliable, even if few known feature points (i.e., less than three) are observed in the images. A new global observability analysis of this tightly-coupled integration is presented to guarantee that the system is observable under the necessary conditions. The analysis conclusions were verified by simulations and field tests. The field tests also indicate that high-precision position (centimeter-level) and attitude (half-degree-level)-integrated solutions can be achieved in a global reference. PMID:25330046

  16. EEG oscillations entrain their phase to high-level features of speech sound.

    PubMed

    Zoefel, Benedikt; VanRullen, Rufin

    2016-01-01

    Phase entrainment of neural oscillations, the brain's adjustment to rhythmic stimulation, is a central component in recent theories of speech comprehension: the alignment between brain oscillations and speech sound improves speech intelligibility. However, phase entrainment to everyday speech sound could also be explained by oscillations passively following the low-level periodicities (e.g., in sound amplitude and spectral content) of auditory stimulation-and not by an adjustment to the speech rhythm per se. Recently, using novel speech/noise mixture stimuli, we have shown that behavioral performance can entrain to speech sound even when high-level features (including phonetic information) are not accompanied by fluctuations in sound amplitude and spectral content. In the present study, we report that neural phase entrainment might underlie our behavioral findings. We observed phase-locking between electroencephalogram (EEG) and speech sound in response not only to original (unprocessed) speech but also to our constructed "high-level" speech/noise mixture stimuli. Phase entrainment to original speech and speech/noise sound did not differ in the degree of entrainment, but rather in the actual phase difference between EEG signal and sound. Phase entrainment was not abolished when speech/noise stimuli were presented in reverse (which disrupts semantic processing), indicating that acoustic (rather than linguistic) high-level features play a major role in the observed neural entrainment. Our results provide further evidence for phase entrainment as a potential mechanism underlying speech processing and segmentation, and for the involvement of high-level processes in the adjustment to the rhythm of speech. PMID:26341026

  17. Weighted simultaneous algebraic reconstruction technique for tomosynthesis imaging of objects with high-attenuation features

    SciTech Connect

    Levakhina, Y. M.; Mueller, J.; Buzug, T. M.; Duschka, R. L.; Vogt, F.; Barkhausen, J.

    2013-03-15

    Purpose: This paper introduces a nonlinear weighting scheme into the backprojection operation within the simultaneous algebraic reconstruction technique (SART). It is designed for tomosynthesis imaging of objects with high-attenuation features in order to reduce limited angle artifacts. Methods: The algorithm estimates which projections potentially produce artifacts in a voxel. The contribution of those projections into the updating term is reduced. In order to identify those projections automatically, a four-dimensional backprojected space representation is used. Weighting coefficients are calculated based on a dissimilarity measure, evaluated in this space. For each combination of an angular view direction and a voxel position an individual weighting coefficient for the updating term is calculated. Results: The feasibility of the proposed approach is shown based on reconstructions of the following real three-dimensional tomosynthesis datasets: a mammography quality phantom, an apple with metal needles, a dried finger bone in water, and a human hand. Datasets have been acquired with a Siemens Mammomat Inspiration tomosynthesis device and reconstructed using SART with and without suggested weighting. Out-of-focus artifacts are described using line profiles and measured using standard deviation (STD) in the plane and below the plane which contains artifact-causing features. Artifacts distribution in axial direction is measured using an artifact spread function (ASF). The volumes reconstructed with the weighting scheme demonstrate the reduction of out-of-focus artifacts, lower STD (meaning reduction of artifacts), and narrower ASF compared to nonweighted SART reconstruction. It is achieved successfully for different kinds of structures: point-like structures such as phantom features, long structures such as metal needles, and fine structures such as trabecular bone structures. Conclusions: Results indicate the feasibility of the proposed algorithm to reduce typical tomosynthesis artifacts produced by high-attenuation features. The proposed algorithm assigns weighting coefficients automatically and no segmentation or tissue-classification steps are required. The algorithm can be included into various iterative reconstruction algorithms with an additive updating strategy. It can also be extended to computed tomography case with the complete set of angular data.

  18. Mechanism of APC/CCDC20 activation by mitotic phosphorylation.

    PubMed

    Qiao, Renping; Weissmann, Florian; Yamaguchi, Masaya; Brown, Nicholas G; VanderLinden, Ryan; Imre, Richard; Jarvis, Marc A; Brunner, Michael R; Davidson, Iain F; Litos, Gabriele; Haselbach, David; Mechtler, Karl; Stark, Holger; Schulman, Brenda A; Peters, Jan-Michael

    2016-05-10

    Chromosome segregation and mitotic exit are initiated by the 1.2-MDa ubiquitin ligase APC/C (anaphase-promoting complex/cyclosome) and its coactivator CDC20 (cell division cycle 20). To avoid chromosome missegregation, APC/C(CDC20) activation is tightly controlled. CDC20 only associates with APC/C in mitosis when APC/C has become phosphorylated and is further inhibited by a mitotic checkpoint complex until all chromosomes are bioriented on the spindle. APC/C contains 14 different types of subunits, most of which are phosphorylated in mitosis on multiple sites. However, it is unknown which of these phospho-sites enable APC/C(CDC20) activation and by which mechanism. Here we have identified 68 evolutionarily conserved mitotic phospho-sites on human APC/C bound to CDC20 and have used the biGBac technique to generate 47 APC/C mutants in which either all 68 sites or subsets of them were replaced by nonphosphorylatable or phospho-mimicking residues. The characterization of these complexes in substrate ubiquitination and degradation assays indicates that phosphorylation of an N-terminal loop region in APC1 is sufficient for binding and activation of APC/C by CDC20. Deletion of the N-terminal APC1 loop enables APC/C(CDC20) activation in the absence of mitotic phosphorylation or phospho-mimicking mutations. These results indicate that binding of CDC20 to APC/C is normally prevented by an autoinhibitory loop in APC1 and that its mitotic phosphorylation relieves this inhibition. The predicted location of the N-terminal APC1 loop implies that this loop controls interactions between the N-terminal domain of CDC20 and APC1 and APC8. These results reveal how APC/C phosphorylation enables CDC20 to bind and activate the APC/C in mitosis. PMID:27114510

  19. Mechanism of APC/CCDC20 activation by mitotic phosphorylation

    PubMed Central

    Qiao, Renping; Weissmann, Florian; Yamaguchi, Masaya; Brown, Nicholas G.; VanderLinden, Ryan; Imre, Richard; Jarvis, Marc A.; Brunner, Michael R.; Davidson, Iain F.; Litos, Gabriele; Haselbach, David; Mechtler, Karl; Stark, Holger; Schulman, Brenda A.; Peters, Jan-Michael

    2016-01-01

    Chromosome segregation and mitotic exit are initiated by the 1.2-MDa ubiquitin ligase APC/C (anaphase-promoting complex/cyclosome) and its coactivator CDC20 (cell division cycle 20). To avoid chromosome missegregation, APC/CCDC20 activation is tightly controlled. CDC20 only associates with APC/C in mitosis when APC/C has become phosphorylated and is further inhibited by a mitotic checkpoint complex until all chromosomes are bioriented on the spindle. APC/C contains 14 different types of subunits, most of which are phosphorylated in mitosis on multiple sites. However, it is unknown which of these phospho-sites enable APC/CCDC20 activation and by which mechanism. Here we have identified 68 evolutionarily conserved mitotic phospho-sites on human APC/C bound to CDC20 and have used the biGBac technique to generate 47 APC/C mutants in which either all 68 sites or subsets of them were replaced by nonphosphorylatable or phospho-mimicking residues. The characterization of these complexes in substrate ubiquitination and degradation assays indicates that phosphorylation of an N-terminal loop region in APC1 is sufficient for binding and activation of APC/C by CDC20. Deletion of the N-terminal APC1 loop enables APC/CCDC20 activation in the absence of mitotic phosphorylation or phospho-mimicking mutations. These results indicate that binding of CDC20 to APC/C is normally prevented by an autoinhibitory loop in APC1 and that its mitotic phosphorylation relieves this inhibition. The predicted location of the N-terminal APC1 loop implies that this loop controls interactions between the N-terminal domain of CDC20 and APC1 and APC8. These results reveal how APC/C phosphorylation enables CDC20 to bind and activate the APC/C in mitosis. PMID:27114510

  20. A Hybrid Feature Subset Selection Algorithm for Analysis of High Correlation Proteomic Data

    PubMed Central

    Kordy, Hussain Montazery; Baygi, Mohammad Hossein Miran; Moradi, Mohammad Hassan

    2012-01-01

    Pathological changes within an organ can be reflected as proteomic patterns in biological fluids such as plasma, serum, and urine. The surface-enhanced laser desorption and ionization time-of-flight mass spectrometry (SELDI-TOF MS) has been used to generate proteomic profiles from biological fluids. Mass spectrometry yields redundant noisy data that the most data points are irrelevant features for differentiating between cancer and normal cases. In this paper, we have proposed a hybrid feature subset selection algorithm based on maximum-discrimination and minimum-correlation coupled with peak scoring criteria. Our algorithm has been applied to two independent SELDI-TOF MS datasets of ovarian cancer obtained from the NCI-FDA clinical proteomics databank. The proposed algorithm has used to extract a set of proteins as potential biomarkers in each dataset. We applied the linear discriminate analysis to identify the important biomarkers. The selected biomarkers have been able to successfully diagnose the ovarian cancer patients from the noncancer control group with an accuracy of 100%, a sensitivity of 100%, and a specificity of 100% in the two datasets. The hybrid algorithm has the advantage that increases reproducibility of selected biomarkers and able to find a small set of proteins with high discrimination power. PMID:23717808

  1. The crystal structure of archaeal serine hydroxymethyltransferase reveals idiosyncratic features likely required to withstand high temperatures.

    PubMed

    Angelucci, Francesco; Morea, Veronica; Angelaccio, Sebastiana; Saccoccia, Fulvio; Contestabile, Roberto; Ilari, Andrea

    2014-12-01

    Serine hydroxymethyltransferases (SHMTs) play an essential role in one-carbon unit metabolism and are used in biomimetic reactions. We determined the crystal structure of free (apo) and pyridoxal-5'-phosphate-bound (holo) SHMT from Methanocaldococcus jannaschii, the first from a hyperthermophile, from the archaea domain of life and that uses H₄MPT as a cofactor, at 2.83 and 3.0 Å resolution, respectively. Idiosyncratic features were observed that are likely to contribute to structure stabilization. At the dimer interface, the C-terminal region folds in a unique fashion with respect to SHMTs from eubacteria and eukarya. At the active site, the conserved tyrosine does not make a cation-π interaction with an arginine like that observed in all other SHMT structures, but establishes an amide-aromatic interaction with Asn257, at a different sequence position. This asparagine residue is conserved and occurs almost exclusively in (hyper)thermophile SHMTs. This led us to formulate the hypothesis that removal of frustrated interactions (such as the Arg-Tyr cation-π interaction occurring in mesophile SHMTs) is an additional strategy of adaptation to high temperature. Both peculiar features may be tested by designing enzyme variants potentially endowed with improved stability for applications in biomimetic processes. PMID:25257552

  2. High Resolution Prediction of Calcium-Binding Sites in 3D Protein Structures Using FEATURE

    PubMed Central

    2015-01-01

    Metal-binding proteins are ubiquitous in biological systems ranging from enzymes to cell surface receptors. Among the various biologically active metal ions, calcium plays a large role in regulating cellular and physiological changes. With the increasing number of high-quality crystal structures of proteins associated with their metal ion ligands, many groups have built models to identify Ca2+ sites in proteins, utilizing information such as structure, geometry, or homology to do the inference. We present a FEATURE-based approach in building such a model and show that our model is able to discriminate between nonsites and calcium-binding sites with a very high precision of more than 98%. We demonstrate the high specificity of our model by applying it to test sets constructed from other ions. We also introduce an algorithm to convert high scoring regions into specific site predictions and demonstrate the usage by scanning a test set of 91 calcium-binding protein structures (190 calcium sites). The algorithm has a recall of more than 93% on the test set with predictions found within 3 Å of the actual sites. PMID:26226489

  3. High-resolution Mapping of Offshore and Onshore Glaciogenic Features in Melville Bay, Northwestern Greenland

    NASA Astrophysics Data System (ADS)

    Freire, F.; Gyllencreutz, R.; Greenwood, S.; Mayer, L. A.; Jakobsson, M.

    2014-12-01

    This study presents results from high resolution mapping in the northwestern part of Greenland's continental shelf, offshore from the Greenland Ice Sheet. The study area is located at about 74o30'N and 58 o40'W where high-resolution seafloor imagery were collected from ~200-500 m water depth. These data were analyzed and compared to existing high-resolution satellite imagery of exposed glacial landforms from the nearby coastal areas. Offshore geophysical mapping equipment consisted of a Kongsberg EM2040 multibeam that was bow-mounted on the sailing vessel Explorer of Sweden together with a Seatex MRU5+ motion sensor and GPS antennas. In addition, a GAVIA autonomous underwater vehicle (AUV) from University of Iceland with installed Geoswath interfometric sonar and Marine Sonic side-scan was used. The data from these systems permitted the production of both 5-m (for the EM2040) and 2-m (for the Geoswath) resolution bathymetric grids for landform analyzes. Sediment characterization analysis was also undertaken using the co-registered backscatter data. The exposed onshore landforms were studied using data from the high-res QuickBird satellite images with a 2-m pixel resolution. Geomorphic analysis of the data shows that past tectonic and glacial scouring processes have shaped the present-day landscape in both the offshore and onshore study areas. The terrain consists of glacially eroded bedrock covered with very thin surficial sediments resembling a 'cnoc-and-lochan' terrain, although the degree of erosion varies spatially, probably as a result of local variations in the rock properties. Different glacially influenced features are identified and described in the study. These features have been used to understand and infer past ice-sheet processes, particularly ice-flow direction and the extent of ice-cover on the continental shelves from previous extreme glaciation events. The backscatter information from the high-resolution interferometric sonar show fine-scale sedimentation patterns which are used to infer bottom water circulation. The study highlights that the use of the high-resolution seafloor mapping systems significantly enhance the quality of geomorphologic landform assessment.

  4. Bayesian Multiscale Analysis of X-Ray Jet Features in High Redshift Quasars

    NASA Astrophysics Data System (ADS)

    McKeough, Kathryn; Siemiginowska, A.; Kashyap, V.; Stein, N.

    2014-01-01

    X-ray emission of powerful quasar jets may be a result of the inverse Compton (IC) process in which the Cosmic Microwave Background (CMB) photons gain energy by interactions with the jet’s relativistic electrons. However, there is no definite evidence that IC/CMB process is responsible for the observed X-ray emission of large scale jets. A step toward understanding the X-ray emission process is to study the Radio and X-ray morphologies of the jet. We implement a sophisticated Bayesian image analysis program, Low-count Image Reconstruction and Analysis (LIRA) (Esch et al. 2004; Conners & van Dyk 2007), to analyze jet features in 11 Chandra images of high redshift quasars (z ~ 2 - 4.8). Out of the 36 regions where knots are visible in the radio jets, nine showed detectable X-ray emission. We measured the ratios of the X-ray and radio luminosities of the detected features and found that they are consistent with the CMB radiation relationship. We derived a range of the bulk lorentz factor (Γ) for detected jet features under the CMB jet emission model. There is no discernible trend of Γ with redshift within the sample. The efficiency of the X-ray emission between the detected jet feature and the corresponding quasar also shows no correlation with redshift. This work is supported in part by the National Science Foundation REU and the Department of Defense ASSURE programs under NSF Grant no.1262851 and by the Smithsonian Institution, and by NASA Contract NAS8-39073 to the Chandra X-ray Center (CXC). This research has made use of data obtained from the Chandra Data Archive and Chandra Source Catalog, and software provided by the CXC in the application packages CIAO, ChIPS, and Sherpa. We thank Teddy Cheung for providing the VLA radio images. Connors, A., & van Dyk, D. A. 2007, Statistical Challenges in Modern Astronomy IV, 371, 101 Esch, D. N., Connors, A., Karovska, M., & van Dyk, D. A. 2004, ApJ, 610, 1213

  5. Design features of a high-intensity, cesium-sputter/plasma-sputter negative ion source

    SciTech Connect

    Alton, G.D.; Mills, G.D.; Dellwo, J.

    1993-12-31

    A versatile, high-intensity, negative ion source has been designed and is now under construction which can be operated in either the cesium-sputter or plasma-sputter mode. The cesium-sputter mode can be effected by installation of a newly designed conical-geometry cesium-surface ionizer; for operation in the plasma-sputter mode, the surface ionizer is removed and either a hot-filament or RIF antenna plasma-discharge igniter is installed. A multicusp magnetic field is specifically provided confining the plasma in the radial direction when the plasma-sputter mode is selected. This arrangement allows comparison of the two modes of operation. Brief descriptions of the design features, ion optics, and anticipated performances of the two source geometries will be presented in this report.

  6. A Composite Approach To The Identification Of High-Level Topological Features In A Histopathologic Image

    NASA Astrophysics Data System (ADS)

    Kuhn, W. P.; Bartels, H. G.; Bartels, P. H.; Richards, D. L.; Saffer, J. S.; Shoemaker, R. L.

    1988-06-01

    Analysis of the large amounts of image data obtainable from very-high-speed scanning laser microscopes places severe demands on computer software and hardware architectures. The automated calculation of features over entire images can provide quantitative data useful to a pathologist who must make a diagnosis. A program that identifies objects of diagnostic interest in an image must utilize a model of the image. An expert system is an effective method for building abstract models of object hierarchies and for utilizing heuristic information. In this paper we discuss a composite approach to image understanding and assessment that utilizes an expert system to control a set of image processing functions for the recognition of various objects in an image.

  7. A Small Mission Featuring an Imaging X-ray Polarimeter with High Sensitivity

    NASA Technical Reports Server (NTRS)

    Weisskopf, Martin C.; Baldini, Luca; Bellazini, Ronaldo; Brez, Alessandro; Costa, Enrico; Dissley, Richard; Elsner, Ronald; Fabiani, Sergio; Matt, Giorgio; Minuti, Massimo; Mulieri, Fabio; O'Dell, Steve; Pinchera, Michelle; Ramsey, Brian; Rubini, Alda; Sgro, Carmelo; Soffitta, Paolo; Spandre, Gloria

    2013-01-01

    We present a detailed description of a small mission capable of obtaining high precision and meaningful measurement of the X-ray polarization of a variety of different classes of cosmic X-ray sources. Compared to other ideas that have been suggested this experiment has demonstrated in the laboratory a number of extremely important features relevant to the ultimate selection of such a mission by a funding agency. The most important of these questions are: 1) Have you demonstrated the sensitivity to a polarized beam at the energies of interest (i.e. the energies which represent the majority (not the minority) of detected photons from the X-ray source of interest? 2) Have you demonstrated that the device's sensitivity to an unpolarized beam is really negligible and/or quantified the impact of any systematic effects upon actual measurements? We present our answers to these questions backed up by laboratory measurements and give an overview of the mission.

  8. MULTIPLE HIGH-VELOCITY SiO MASER FEATURES FROM THE HIGH-MASS PROTOSTAR W51 NORTH

    SciTech Connect

    Cho, Se-Hyung; Kim, Jaeheon; Byun, Do-Young E-mail: jhkim@kasi.re.kr

    2011-02-01

    We present the detection of multiple high-velocity silicon monoxide (SiO v = 1, 2, J = 1-0) maser features in the high-mass protostar W51 North which are distributed over an exceedingly large velocity range from 105 to 230 km s{sup -1}. The SiO v = 1, J = 1-0 maser emission shows 3-5 narrow components which span a velocity range from 154 to 230 km s{sup -1} according to observational epochs. The SiO v = 2, J = 1-0 maser also shows 3-5 narrow components that do not correspond to the SiO v = 1 maser and span a velocity range from 105 to 154 km s{sup -1}. The multiple maser components show significant changes on very short timescales (<1 month) from epoch to epoch. We suggest that the high-velocity SiO masers may be emanated from massive star-forming activity of the W51 North protostar as SiO maser jets and will be a good probe of the earliest evolutionary stages of high-mass star formation via an accretion model. Further high angular resolution observations will be required for confirmation.

  9. Mitotic chromosome loss in a radiation-sensitive strain of the yeast Saccharomyces cerevisiae

    SciTech Connect

    Mortimer, R.K.; Contopoulou, R.; Schild, D.

    1981-09-01

    Cells of Saccharomyces cerevisiae with mutations in the RAD52 gene have previously been shown to be defective in meiotic and mitotic recombination, in sporulation, and in repair of radiation-induced damage to DNA. In this study we show that diploid cells homozygous for rad52 lose chromosomes at high frequencies and that these frequencies of loss can be increased dramatically by exposure of these cells to x-rays. Genetic analyses of survivors of x-ray treatment demonstrate that chromosome loss events result in the conversion of diploid cells to cells with near haploid chromosome numbers.

  10. Design features and performance of the LAMPF high-intensity beam area

    SciTech Connect

    Agnew, L.; Grisham, D.; Macek, R.J.; Sommer, W.F.; Werbeck, R.D.

    1983-01-01

    LAMPF is a multi-purpose high-intensity meson factory capable of producing a 1 mA beam of 800-MeV protons. The three target cells and the beam stop facilities in the high intensity area have many special design features that are required for operation in the presence of high heat loads and intense radiation fields where accessibility is extremely limited. Reliable targets, beam windows, beam stops, beam transport and diagnostic components, vacuum enclosures, and auxiliary systems have been developed. Sophisticated remote-handling systems are employed for maintenance. Complex protection systems have been developed to guard against damage caused by errant beam. Beam availability approaching 90% has been achieved at currents of 600 to 700 ..mu..A. A new facility for direct proton and neutron radiation effects studies will be installed in 1985. The new facility will provide an integrated spallation neutron flux of up to 5 x 10/sup 17/ m/sup -2/s/sup -1/ and will anable proton irradiation studies in the primary beam.

  11. Prioritizing spatial accuracy in high-resolution fMRI data using multivariate feature weight mapping

    PubMed Central

    Buschmann, Tilo; Lohmann, Gabriele; Margulies, Daniel S.; Trampel, Robert; Turner, Robert

    2014-01-01

    Although ultra-high-field fMRI at field strengths of 7T or above provides substantial gains in BOLD contrast-to-noise ratio, when very high-resolution fMRI is required such gains are inevitably reduced. The improvement in sensitivity provided by multivariate analysis techniques, as compared with univariate methods, then becomes especially welcome. Information mapping approaches are commonly used, such as the searchlight technique, which take into account the spatially distributed patterns of activation in order to predict stimulus conditions. However, the popular searchlight decoding technique, in particular, has been found to be prone to spatial inaccuracies. For instance, the spatial extent of informative areas is generally exaggerated, and their spatial configuration is distorted. We propose the combination of a non-parametric and permutation-based statistical framework with linear classifiers. We term this new combined method Feature Weight Mapping (FWM). The main goal of the proposed method is to map the specific contribution of each voxel to the classification decision while including a correction for the multiple comparisons problem. Next, we compare this new method to the searchlight approach using a simulation and ultra-high-field 7T experimental data. We found that the searchlight method led to spatial inaccuracies that are especially noticeable in high-resolution fMRI data. In contrast, FWM was more spatially precise, revealing both informative anatomical structures as well as the direction by which voxels contribute to the classification. By maximizing the spatial accuracy of ultra-high-field fMRI results, global multivariate methods provide a substantial improvement for characterizing structure-function relationships. PMID:24795548

  12. Extraction of Airport Features from High Resolution Satellite Imagery for Design and Risk Assessment

    NASA Technical Reports Server (NTRS)

    Robinson, Chris; Qiu, You-Liang; Jensen, John R.; Schill, Steven R.; Floyd, Mike

    2001-01-01

    The LPA Group, consisting of 17 offices located throughout the eastern and central United States is an architectural, engineering and planning firm specializing in the development of Airports, Roads and Bridges. The primary focus of this ARC project is concerned with assisting their aviation specialists who work in the areas of Airport Planning, Airfield Design, Landside Design, Terminal Building Planning and design, and various other construction services. The LPA Group wanted to test the utility of high-resolution commercial satellite imagery for the purpose of extracting airport elevation features in the glide path areas surrounding the Columbia Metropolitan Airport. By incorporating remote sensing techniques into their airport planning process, LPA wanted to investigate whether or not it is possible to save time and money while achieving the equivalent accuracy as traditional planning methods. The Affiliate Research Center (ARC) at the University of South Carolina investigated the use of remotely sensed imagery for the extraction of feature elevations in the glide path zone. A stereo pair of IKONOS panchromatic satellite images, which has a spatial resolution of 1 x 1 m, was used to determine elevations of aviation obstructions such as buildings, trees, towers and fence-lines. A validation dataset was provided by the LPA Group to assess the accuracy of the measurements derived from the IKONOS imagery. The initial goal of this project was to test the utility of IKONOS imagery in feature extraction using ERDAS Stereo Analyst. This goal was never achieved due to problems with ERDAS software support of the IKONOS sensor model and the unavailability of imperative sensor model information from Space Imaging. The obstacles encountered in this project pertaining to ERDAS Stereo Analyst and IKONOS imagery will be reviewed in more detail later in this report. As a result of the technical difficulties with Stereo Analyst, ERDAS OrthoBASE was used to derive aviation obstruction measurements for this project. After collecting ancillary data such as GPS locations, South Carolina Geodetic Survey and Aero Dynamics ground survey points to set up the OrthoBASE Block File, measurements were taken of the various glide path obstructions and compared to the validation dataset. This process yielded the following conclusions: The IKONOS stereo model in conjunction with Imagine OrthoBASE can provide The LPA Group with a fast and cost efficient method for assessing aviation obstructions. Also, by creating our own stereo model we achieved any accuracy better currently available commercial products.

  13. Retrieval Using Texture Features in High Resolution Multi-spectral Satellite Imagery

    SciTech Connect

    Newsam, S D; Kamath, C

    2004-01-22

    Texture features have long been used in remote sensing applications to represent and retrieve image regions similar to a query region. Various representations of texture have been proposed based on the Fourier power spectrum, spatial co-occurrence, wavelets, Gabor filters, etc. These representations vary in their computational complexity and their suitability for representing different region types. Much of the work done thus far has focused on panchromatic imagery at low to moderate spatial resolutions, such as images from Landsat 1-7 which have a resolution of 15-30 m/pixel, and from SPOT 1-5 which have a resolution of 2.5-20 m/pixel. However, it is not clear which texture representation works best for the new classes of high resolution panchromatic (60-100 cm/pixel) and multi-spectral (4 bands for red, green, blue, and near infra-red at 2.4-4 m/pixel) imagery. It is also not clear how the different spectral bands should be combined. In this paper, we investigate the retrieval performance of several different texture representations using multi-spectral satellite images from IKONOS. A query-by-example framework, along with a manually chosen ground truth dataset, allows different combinations of texture representations and spectral bands to be compared. We focus on the specific problem of retrieving inhabited regions from images of urban and rural scenes. Preliminary results show that (1) the use of all spectral bands improves the retrieval performance, and (2) co-occurrence, wavelet and Gabor texture features perform comparably.

  14. Clinical Application of High-Resolution Computed Tomographic Imaging Features of Community-Acquired Pneumonia.

    PubMed

    Nie, Yunqiang; Li, Cuiyun; Zhang, Jingling; Wang, Hui; Han, Ping; Lv, Xin; Xu, Xinyi; Guo, Miao

    2016-01-01

    BACKGROUND This article discusses the value of high-resolution computed tomography (HRCT) in the diagnosis and treatment of pulmonary infections. Lung infection caused by pathogens is an important cause of death. Traditional methods to treat lung infection involved empirical antibiotic therapy. Thin-slice CT scanning is widely used in the clinical setting, and HRCT scan can very clearly show alveolar and bronchiolar involvement of infection. MATERIAL AND METHODS In total, 178 patients with community-acquired pneumonia (CAP) were enrolled. All the patients underwent CT scan, qualified sputum, and blood samples for culture or immunological biochemical tests. CT imaging features, pathogenic bacteria, and treatment results were used for statistical analysis. RESULTS In 77 patients with lobar consolidation, the rate of detection was 43.26% (77/178), and in 101 patients with lobular pneumonia it was 56.74% (101/178). In 51 patients, pathogenic bacteria were detected (28.65%, 51/178). Sixteen of 33 patients detected with bacteria had cavities (48.5%, 16/33) and 35 of 145 patients detected with bacteria had no cavities (24.1%, 35/145). The difference between the 2 groups was statistically significant (χ2=7.795, P=0.005). According to the pathogenic bacteria, 38 patients were cured (74.51%, 38/51), and according to the CT imaging features 81 patients were cured (71.05%, 81/114). No statistically significant difference was found between them (χ2=0.209, P=0.647). CONCLUSIONS Treatment effect of CAP based on HRCT findings is not inferior to treatment effect guided by microbial characterization. PMID:27031210

  15. Clinical Application of High-Resolution Computed Tomographic Imaging Features of Community-Acquired Pneumonia

    PubMed Central

    Nie, Yunqiang; Li, Cuiyun; Zhang, Jingling; Wang, Hui; Han, Ping; Lv, Xin; Xu, Xinyi; Guo, Miao

    2016-01-01

    Background This article discusses the value of high-resolution computed tomography (HRCT) in the diagnosis and treatment of pulmonary infections. Lung infection caused by pathogens is an important cause of death. Traditional methods to treat lung infection involved empirical antibiotic therapy. Thin-slice CT scanning is widely used in the clinical setting, and HRCT scan can very clearly show alveolar and bronchiolar involvement of infection. Material/Methods In total, 178 patients with community-acquired pneumonia (CAP) were enrolled. All the patients underwent CT scan, qualified sputum, and blood samples for culture or immunological biochemical tests. CT imaging features, pathogenic bacteria, and treatment results were used for statistical analysis. Results In 77 patients with lobar consolidation, the rate of detection was 43.26% (77/178), and in 101 patients with lobular pneumonia it was 56.74% (101/178). In 51 patients, pathogenic bacteria were detected (28.65%, 51/178). Sixteen of 33 patients detected with bacteria had cavities (48.5%, 16/33) and 35 of 145 patients detected with bacteria had no cavities (24.1%, 35/145). The difference between the 2 groups was statistically significant (χ2=7.795, P=0.005). According to the pathogenic bacteria, 38 patients were cured (74.51%, 38/51), and according to the CT imaging features 81 patients were cured (71.05%, 81/114). No statistically significant difference was found between them (χ2=0.209, P=0.647). Conclusions Treatment effect of CAP based on HRCT findings is not inferior to treatment effect guided by microbial characterization. PMID:27031210

  16. Detailed Hydrographic Feature Extraction from High-Resolution LiDAR Data

    SciTech Connect

    Danny L. Anderson

    2012-05-01

    Detailed hydrographic feature extraction from high-resolution light detection and ranging (LiDAR) data is investigated. Methods for quantitatively evaluating and comparing such extractions are presented, including the use of sinuosity and longitudinal root-mean-square-error (LRMSE). These metrics are then used to quantitatively compare stream networks in two studies. The first study examines the effect of raster cell size on watershed boundaries and stream networks delineated from LiDAR-derived digital elevation models (DEMs). The study confirmed that, with the greatly increased resolution of LiDAR data, smaller cell sizes generally yielded better stream network delineations, based on sinuosity and LRMSE. The second study demonstrates a new method of delineating a stream directly from LiDAR point clouds, without the intermediate step of deriving a DEM. Direct use of LiDAR point clouds could improve efficiency and accuracy of hydrographic feature extractions. The direct delineation method developed herein and termed “mDn”, is an extension of the D8 method that has been used for several decades with gridded raster data. The method divides the region around a starting point into sectors, using the LiDAR data points within each sector to determine an average slope, and selecting the sector with the greatest downward slope to determine the direction of flow. An mDn delineation was compared with a traditional grid-based delineation, using TauDEM, and other readily available, common stream data sets. Although, the TauDEM delineation yielded a sinuosity that more closely matches the reference, the mDn delineation yielded a sinuosity that was higher than either the TauDEM method or the existing published stream delineations. Furthermore, stream delineation using the mDn method yielded the smallest LRMSE.

  17. The Endocrine Dyscrasia that Accompanies Menopause and Andropause Induces Aberrant Cell Cycle Signaling that Triggers Cell Cycle Reentry of Post-mitotic Neurons, Neurodysfunction, Neurodegeneration and Cognitive Disease

    PubMed Central

    Atwood, Craig S.; Bowen, Richard L.

    2016-01-01

    Sex hormones are the physiological factors that regulate neurogenesis during embryogenesis and continuing through adulthood. These hormones support the formation of brain structures such as dendritic spines, axons and synapses required for the capture of information (memories). Intriguingly, a recent animal study has demonstrated that induction of neurogenesis results in the loss of previously encoded memories in animals (e.g. infantile amnesia). In this connection, much evidence now indicates that Alzheimer’s disease (AD) also involves aberrant re-entry of post-mitotic neurons into the cell cycle. Cell cycle abnormalities appear very early in the disease, prior to the appearance of plaques and tangles, and explain the biochemical, neuropathological and cognitive changes observed with disease progression. Since sex hormones control when and how neurons proliferate and differentiate, the endocrine dyscrasia that accompanies menopause and andropause is a key signaling event that impacts neurogenesis and the acquisition, processing, storage and recall of memories. Here we review the biochemical, epidemiological and clinical evidence that alterations in endocrine signaling with menopause and andropause drive the aberrant re-entry of post-mitotic neurons into an abortive cell cycle with neurite retraction that leads to neuron dysfunction and death. When the reproductive axis is in balance, luteinizing hormone (LH), and its fetal homolog, human chorionic gonadotropin (hCG), promote pluripotent human and totipotent murine embryonic stem cell and neuron proliferation. However, strong evidence supports menopausal/andropausal elevations in the ratio of LH:sex steroids as driving aberrant mitotic events mediated by the upregulation of tumor necrosis factor, amyloid-β precursor protein processing towards the production of mitogenic Aβ, and the activation of Cdk5, a key regulator of cell cycle progression and tau phosphorylation (a cardinal feature of both neurogenesis and neurodegeneration). Cognitive studies also demonstrate the negative consequences of a high LH:sex steroid ratio on human cognitive performance. Prospective epidemiological and clinical evidence in humans supports lowering the ratio of circulating gonadotropins-GnRH to sex steroids in reducing the incidence of AD and halting cognitive decline. Together, these data support endocrine dyscrasia and the subsequent loss of cell cycle control as an important etiological event in the development of neurodegenerative diseases including AD, stroke and Parkinson’s disease. PMID:26188949

  18. The anthelmintic drug mebendazole induces mitotic arrest and apoptosis by depolymerizing tubulin in non-small cell lung cancer cells.

    PubMed

    Sasaki, Ji-ichiro; Ramesh, Rajagopal; Chada, Sunil; Gomyo, Yoshihito; Roth, Jack A; Mukhopadhyay, Tapas

    2002-11-01

    Microtubules have a critical role in cell division, and consequently various microtubule inhibitors have been developed as anticancer drugs. In this study, we assess mebendazole (MZ), a microtubule-disrupting anthelmintic that exhibits a potent antitumor property both in vitro and in vivo. Treatment of lung cancer cell lines with MZ caused mitotic arrest, followed by apoptotic cell death with the feature of caspase activation and cytochrome c release. MZ induces abnormal spindle formation in mitotic cancer cells and enhances the depolymerization of tubulin, but the efficacy of depolymerization by MZ is lower than that by nocodazole. Oral administration of MZ in mice elicited a strong antitumor effect in a s.c. model and reduced lung colonies in experimentally induced lung metastasis without any toxicity when compared with paclitaxel-treated mice. We speculate that tumor cells may be defective in one mitotic checkpoint function and sensitive to the spindle inhibitor MZ. Abnormal spindle formation may be the key factor determining whether a cell undergoes apoptosis, whereas strong microtubule inhibitors elicit toxicity even in normal cells. PMID:12479701

  19. Architectural epigenetics: mitotic retention of mammalian transcriptional regulatory information.

    PubMed

    Zaidi, Sayyed K; Young, Daniel W; Montecino, Martin; Lian, Jane B; Stein, Janet L; van Wijnen, Andre J; Stein, Gary S

    2010-10-01

    Epigenetic regulatory information must be retained during mammalian cell division to sustain phenotype-specific and physiologically responsive gene expression in the progeny cells. Histone modifications, DNA methylation, and RNA-mediated silencing are well-defined epigenetic mechanisms that control the cellular phenotype by regulating gene expression. Recent results suggest that the mitotic retention of nuclease hypersensitivity, selective histone marks, as well as the lineage-specific transcription factor occupancy of promoter elements contribute to the epigenetic control of sustained cellular identity in progeny cells. We propose that these mitotic epigenetic signatures collectively constitute architectural epigenetics, a novel and essential mechanism that conveys regulatory information to sustain the control of phenotype and proliferation in progeny cells by bookmarking genes for activation or suppression. PMID:20696837

  20. Molecular pathways regulating mitotic spindle orientation in animal cells

    PubMed Central

    Lu, Michelle S.; Johnston, Christopher A.

    2013-01-01

    Orientation of the cell division axis is essential for the correct development and maintenance of tissue morphology, both for symmetric cell divisions and for the asymmetric distribution of fate determinants during, for example, stem cell divisions. Oriented cell division depends on the positioning of the mitotic spindle relative to an axis of polarity. Recent studies have illuminated an expanding list of spindle orientation regulators, and a molecular model for how cells couple cortical polarity with spindle positioning has begun to emerge. Here, we review both the well-established spindle orientation pathways and recently identified regulators, focusing on how communication between the cell cortex and the spindle is achieved, to provide a contemporary view of how positioning of the mitotic spindle occurs. PMID:23571210

  1. Brownian dynamics simulation of fission yeast mitotic spindle formation

    NASA Astrophysics Data System (ADS)

    Edelmaier, Christopher

    2014-03-01

    The mitotic spindle segregates chromosomes during mitosis. The dynamics that establish bipolar spindle formation are not well understood. We have developed a computational model of fission-yeast mitotic spindle formation using Brownian dynamics and kinetic Monte Carlo methods. Our model includes rigid, dynamic microtubules, a spherical nuclear envelope, spindle pole bodies anchored in the nuclear envelope, and crosslinkers and crosslinking motor proteins. Crosslinkers and crosslinking motor proteins attach and detach in a grand canonical ensemble, and exert forces and torques on the attached microtubules. We have modeled increased affinity for crosslinking motor attachment to antiparallel microtubule pairs, and stabilization of microtubules in the interpolar bundle. We study parameters controlling the stability of the interpolar bundle and assembly of a bipolar spindle from initially adjacent spindle-pole bodies.

  2. Application Prospects and Microstructural Features in Laser-Induced Rapidly Solidified High-Entropy Alloys

    NASA Astrophysics Data System (ADS)

    Zhang, Hui; Pan, Ye; He, Yi-Zhu; Wu, Ji-Li; Yue, T. M.; Guo, Sheng

    2014-10-01

    Recently, high-entropy alloys (HEAs) have attracted much interest in the materials community, as they offer massive opportunities to observe new phenomena, explore new structure, and develop new materials. Particularly, it is attractive to prepare high-performance HEA coatings by laser-induced rapid solidification, which can be formed on the surface of components and parts in a variety of sizes and shapes with a lower cost in comparison with those bulk material fabrication methods. From the technical point of view, laser-induced rapid solidification could hamper the compositional segregation, improve the solubility in solid-solution phases, and lead to the strengthening effect by the grain refinement. This article reviews the recent work on the typical microstructural features and the mechanical and chemical properties in laser-induced rapidly solidified HEAs, and these data are compared with conventional Co- and Ni-based alloy coatings. The article concludes with suggestions for future research and development in HEAs, from considerations of their characteristic properties.

  3. An efficient realtime video compression algorithm with high feature preserving capability

    NASA Astrophysics Data System (ADS)

    Al-Jawad, Naseer; Ehlers, Johan; Jassim, Sabah

    2006-05-01

    Mobile Phones and other hand held devices are constrained in their memory and computational power, and yet new generations of theses devices provide access to the web-based services and are equipped with digital cameras that make them more attractive to users. These added capabilities are expected to help incorporate such devices into the global communication system. In order to take advantage of these capabilities, there are desperate need for highly efficient algorithms including real-time image and video processing and transmission. This paper is concerned with high quality video compression for constrained mobile devices. We attempt to tweak a wavelet-based feature-preserving image compression technique that we have developed recently, so as to make it suitable for implementation on mobile phones and PDA's. The earlier version of the compression algorithm exploits the statistical properties of the multi-resolution wavelet-transformed images. The main modification is based on the observation that in many cases the statistical parameters of wavelet subbands of adjacent video frames do not differ significantly. We shall investigate the possibility of re-using codebooks for a sequence of adjacent frames without having adverse effect on image quality if any. Such an approach results in significant bandwidth and processing-time savings. The performance of this scheme will be tested in comparison to other video compression methods. Such a scheme is expected to be of use in security applications such as transmission of biometric data for a server-based verification.

  4. High altitude pulmonary edema-clinical features, pathophysiology, prevention and treatment

    PubMed Central

    Paralikar, Swapnil J.

    2012-01-01

    High altitude pulmonary edema (HAPE) is a noncardiogenic pulmonary edema which typically occurs in lowlanders who ascend rapidly to altitudes greater than 2500-3000 m. Early symptoms of HAPE include a nonproductive cough, dyspnoea on exertion and reduced exercise performance. Later, dyspnoea occurs at rest. Clinical features are cyanosis, tachycardia, tachypnoea and elevated body temperature generally not exceeding 38.5°C. Rales are discrete initially and located over the middle lung fields. HAPE mainly occurs due to exaggerated hypoxic pulmonary vasoconstriction and elevated pulmonary artery pressure. It has been observed that HAPE is a high permeability type of edema occurring also due to leaks in the capillary wall (‘stress failure’). Slow descent is the most effective method for prevention; in addition, graded ascent and time for acclimatization, low sleeping altitudes, avoidance of alcohol and sleeping pills, and avoidance of exercise are the key to preventing HAPE. Treatment of HAPE consists of immediate improvement of oxygenation either by supplemental oxygen, hyperbaric treatment, or by rapid descent. PMID:23580834

  5. Mitotic activity in dorsal epidermis of Rana pipiens.

    NASA Technical Reports Server (NTRS)

    Garcia-Arce, H.; Mizell, S.

    1972-01-01

    Study of statistically significant rhythms of mitotic division in dorsal epidermis of frogs, Rana pipiens, exposed to a 12:12 light:dark environment for 14 days. The results include the findings that (1) male animals have a primary period of 22 hr in summer and 18 hr in winter, (2) female animals have an 18 hr period, and (3) parapinealectomy and blinding abolish the rhythm.

  6. Pattern formation in stochastic systems: Magnetized billiards and mitotic spindles

    NASA Astrophysics Data System (ADS)

    Schaffner, Stuart C.

    Physical systems that exhibit chaotic behavior or are subject to thermal noise are treated as random processes, especially if the state of the system cannot be measured precisely. Here we examine two such systems. The first is a single electron confined to a wedge-shaped section of a disk, called a billiard, in the presence of a uniform transverse magnetic field. The system exhibits a mixture of chaotic and nonchaotic behavior at different values of the magnetic field strength. If the size of the billiard is on the order of micrometers, as in a quantum dot, both quantum and classical analyses are necessary. The second system is a collection of stiff fibers, called microtubules, suspended in a fluid called the cytoplasm, and lying over chromosomes in a cell. The cytoplasm supplies molecular motors and fuel for the motors. The chromosomes supply motor attachment points. The combination causes the microtubules to self-assemble into a coherent structure called the mitotic spindle. This structure is vital to cell division in plants and animals. Elements of the mitotic spindle have sizes ranging from nanometers to micrometers, and all are subject to considerable thermal agitation. Mitotic spindle self-assembly occurs despite the randomizing effect of this thermal motion. We studied both systems by constructing physical models described by mathematical equations. From these we were able to perform computer simulations. For the billiard problem, we made innovative use of geometric symmetries. These symmetries allowed us to construct efficient representations of both classical and quantum systems. We found a new region of integrable trajectories for a magnetic field above that required to produce completely chaotic orbits. For the mitotic spindle, we were the first to demonstrate spindle self-assembly in a model that matches conditions reported by experimental biologists. Our simulations have shed significant light on which of the many elements in this complex system are vital for spindle formation.

  7. High-Resolution Infrared Space Observatory Spectroscopy of the Unidentified 21 Micron Feature

    NASA Technical Reports Server (NTRS)

    Volk, Kevin; Kwok, Sun; Hrivnak, Bruce

    1999-01-01

    We present Infrared Space Observatory SWS06 mode observations of the 21 micron feature in eight sources, including a first "detection of the feature in IRAS Z02229+6208. The observed feature peak-to-continuum ratios range from 0.13 in IRAS Z02229+6208 to 1.30 in IRAS 07134+1005. The normalized spectra, obtained by the removal of the underlying continua and by scaling the features to the same peak flux value. show that all features have the same intrinsic profile and peak wavelength. There is no evidence for any discrete substructure due to molecular bands in the observed spectra, suggesting that the 21 micron feature is due to either a solid substance or a mixture of many similarly structured large molecules.

  8. Distance-Independence of Mitotic Intrachromosomal Recombination in Saccharomyces Cerevisiae

    PubMed Central

    Yuan, L. W.; Keil, R. L.

    1990-01-01

    Many genetic studies have shown that the frequency of homologous recombination depends largely on the distance in which recombination can occur. We have studied the effect of varying the length of duplicated sequences on the frequency of mitotic intrachromosomal recombination in Saccharomyces cerevisiae. We find that the frequency of recombination resulting in the loss of one of the repeats and the intervening sequences reaches a plateau when the repeats are short. In addition, the frequency of recombination to correct a point mutation contained in one of these repeats is not proportional to the size of the duplication but rather depends dramatically on the location of the mutation within the repeated sequences. However, the frequency of mitotic interchromosomal reciprocal recombination is dependent on the distance separating the markers. The difference in the response of intrachromosomal and interchromosomal mitotic recombination to increasing lengths of homology may indicate there are different rate-limiting steps for recombination in these two cases. These findings have important implications for the maintenance and evolution of duplicated sequences. PMID:2407612

  9. Role of senescence and mitotic catastrophe in cancer therapy

    PubMed Central

    2010-01-01

    Senescence and mitotic catastrophe (MC) are two distinct crucial non-apoptotic mechanisms, often triggered in cancer cells and tissues in response to anti-cancer drugs. Chemotherapeuticals and myriad other factors induce cell eradication via these routes. While senescence drives the cells to a state of quiescence, MC drives the cells towards death during the course of mitosis. The senescent phenotype distinguishes tumor cells that survived drug exposure but lost the ability to form colonies from those that recover and proliferate after treatment. Although senescent cells do not proliferate, they are metabolically active and may secrete proteins with potential tumor-promoting activities. The other anti-proliferative response of tumor cells is MC that is a form of cell death that results from abnormal mitosis and leads to the formation of interphase cells with multiple micronuclei. Different classes of cytotoxic agents induce MC, but the pathways of abnormal mitosis differ depending on the nature of the inducer and the status of cell-cycle checkpoints. In this review, we compare the two pathways and mention that they are activated to curb the growth of tumors. Altogether, we have highlighted the possibilities of the use of senescence targeting drugs, mitotic kinases and anti-mitotic agents in fabricating novel strategies in cancer control. PMID:20205872

  10. Mitotic Exit in the Absence of Separase Activity

    PubMed Central

    Lu, Ying

    2009-01-01

    In budding yeast, three interdigitated pathways regulate mitotic exit (ME): mitotic cyclin–cyclin-dependent kinase (Cdk) inactivation; the Cdc14 early anaphase release (FEAR) network, including a nonproteolytic function of separase (Esp1); and the mitotic exit network (MEN) driven by interaction between the spindle pole body and the bud cortex. Here, we evaluate the contributions of these pathways to ME kinetics. Reducing Cdk activity is critical for ME, and the MEN contributes strongly to ME efficiency. Esp1 contributes to ME kinetics mainly through cohesin cleavage: the Esp1 requirement can be largely bypassed if cells are provided Esp1-independent means of separating sister chromatids. In the absence of Esp1 activity, we observed only a minor ME delay consistent with a FEAR defect. Esp1 overexpression drives ME in Cdc20-depleted cells arrested in metaphase. We have found that this activity of overexpressed Esp1 depended on spindle integrity and the MEN. We defined the first quantitative measure for Cdc14 release based on colocalization with the Net1 nucleolar anchor. This measure indicates efficient Cdc14 release upon MEN activation; release driven by Esp1 in the absence of microtubules was inefficient and incapable of driving ME. We also found a novel role for the MEN: activating Cdc14 nuclear export, even in the absence of Net1. PMID:19144818

  11. Pulling it together: The mitotic function of TACC3.

    PubMed

    Hood, Fiona E; Royle, Stephen J

    2011-05-01

    Transforming acidic coiled coil 3 (TACC3) is a non-motor microtubule-associated protein (MAP) that is important for mitotic spindle stability and organization. The exact mechanism by which TACC3 acts at microtubules to stabilize the spindle has been unclear. However, several recent studies identified that the TACC3 complex at microtubules contains clathrin in addition to its previously identified binding partner, colonic and hepatic tumor overexpressed gene (ch-TOG). In this complex, phosphorylated TACC3 interacts directly with both ch-TOG and clathrin heavy chain, promoting accumulation of all complex members at the mitotic spindle. This complex stabilizes kinetochore fibers within the spindle by forming cross-bridges that link adjacent microtubules in these bundles. So, TACC3 is an adaptor that recruits ch-TOG and clathrin to mitotic microtubules, in an Aurora A kinase-regulated manner. In this mini-review we will describe the recent advances in the understanding of TACC 3 function and present a model that pulls together these new data with previous observations. PMID:21922039

  12. The Plk1-dependent Phosphoproteome of the Early Mitotic Spindle*

    PubMed Central

    Santamaria, Anna; Wang, Bin; Elowe, Sabine; Malik, Rainer; Zhang, Feng; Bauer, Manuel; Schmidt, Alexander; Silljé, Herman H. W.; Körner, Roman; Nigg, Erich A.

    2011-01-01

    Polo-like kinases regulate many aspects of mitotic and meiotic progression from yeast to man. In early mitosis, mammalian Polo-like kinase 1 (Plk1) controls centrosome maturation, spindle assembly, and microtubule attachment to kinetochores. However, despite the essential and diverse functions of Plk1, the full range of Plk1 substrates remains to be explored. To investigate the Plk1-dependent phosphoproteome of the human mitotic spindle, we combined stable isotope labeling by amino acids in cell culture with Plk1 inactivation or depletion followed by spindle isolation and mass spectrometry. Our study identified 358 unique Plk1-dependent phosphorylation sites on spindle proteins, including novel substrates, illustrating the complexity of the Plk1-dependent signaling network. Over 100 sites were validated by in vitro phosphorylation of peptide arrays, resulting in a broadening of the Plk1 consensus motif. Collectively, our data provide a rich source of information on Plk1-dependent phosphorylation, Plk1 docking to substrates, the influence of phosphorylation on protein localization, and the functional interaction between Plk1 and Aurora A on the early mitotic spindle. PMID:20860994

  13. Mitotic recombination and uniparental disomy in Beckwith-Wiedemann syndrome.

    PubMed

    Cooper, Wendy N; Curley, Rebecca; Macdonald, Fiona; Maher, Eamonn R

    2007-05-01

    Beckwith-Wiedemann syndrome (BWS) is a model human imprinting disorder resulting from altered activity of one or more genes in the 11p15.5 imprinted gene cluster. Approximately 20% of BWS cases have uniparental disomy (UPD) of chromosome 11. Such cases appear to result from mitotic recombination occurring in early embryogenesis and offer a rare opportunity to study mitotic recombination in nonneoplastic cells. We analyzed a cohort of 52 children with BWS and UPD using a panel of microsatellite markers for chromosome 11. All cases demonstrated mosaic paternal isodisomy, and IGF2 and H19 were included in the segment of UPD in all cases. However, the extent of segmental disomy was variable, with no evidence of clustering of the proximal UPD breakpoint. In most cases (92% of those informative) UPD did not involve 11q, but 4 patients demonstrated UPD for the whole of chromosome 11. In contrast to meiotic recombination, the mitotic recombination frequency did not decline near the centromere. PMID:17337339

  14. Feature selection for neural network based defect classification of ceramic components using high frequency ultrasound.

    PubMed

    Kesharaju, Manasa; Nagarajah, Romesh

    2015-09-01

    The motivation for this research stems from a need for providing a non-destructive testing method capable of detecting and locating any defects and microstructural variations within armour ceramic components before issuing them to the soldiers who rely on them for their survival. The development of an automated ultrasonic inspection based classification system would make possible the checking of each ceramic component and immediately alert the operator about the presence of defects. Generally, in many classification problems a choice of features or dimensionality reduction is significant and simultaneously very difficult, as a substantial computational effort is required to evaluate possible feature subsets. In this research, a combination of artificial neural networks and genetic algorithms are used to optimize the feature subset used in classification of various defects in reaction-sintered silicon carbide ceramic components. Initially wavelet based feature extraction is implemented from the region of interest. An Artificial Neural Network classifier is employed to evaluate the performance of these features. Genetic Algorithm based feature selection is performed. Principal Component Analysis is a popular technique used for feature selection and is compared with the genetic algorithm based technique in terms of classification accuracy and selection of optimal number of features. The experimental results confirm that features identified by Principal Component Analysis lead to improved performance in terms of classification percentage with 96% than Genetic algorithm with 94%. PMID:26081920

  15. Universal features in the photoemission spectroscopy of high-temperature superconductors.

    PubMed

    Zhao, Junjing; Chatterjee, Utpal; Ai, Dingfei; Hinks, David G; Zheng, Hong; Gu, G D; Castellan, John-Paul; Rosenkranz, Stephan; Claus, Helmut; Norman, Michael R; Randeria, Mohit; Campuzano, Juan Carlos

    2013-10-29

    The energy gap for electronic excitations is one of the most important characteristics of the superconducting state, as it directly reflects the pairing of electrons. In the copper-oxide high-temperature superconductors (HTSCs), a strongly anisotropic energy gap, which vanishes along high-symmetry directions, is a clear manifestation of the d-wave symmetry of the pairing. There is, however, a dramatic change in the form of the gap anisotropy with reduced carrier concentration (underdoping). Although the vanishing of the gap along the diagonal to the square Cu-O bond directions is robust, the doping dependence of the large gap along the Cu-O directions suggests that its origin might be different from pairing. It is thus tempting to associate the large gap with a second-order parameter distinct from superconductivity. We use angle-resolved photoemission spectroscopy to show that the two-gap behavior and the destruction of well-defined electronic excitations are not universal features of HTSCs, and depend sensitively on how the underdoped materials are prepared. Depending on cation substitution, underdoped samples either show two-gap behavior or not. In contrast, many other characteristics of HTSCs, such as the dome-like dependence of on doping, long-lived excitations along the diagonals to the Cu-O bonds, and an energy gap at the Brillouin zone boundary that decreases monotonically with doping while persisting above (the pseudogap), are present in all samples, irrespective of whether they exhibit two-gap behavior or not. Our results imply that universal aspects of high- superconductivity are relatively insensitive to differences in the electronic states along the Cu-O bond directions. PMID:24101464

  16. Universal features in the photoemission spectroscopy of high-temperature superconductors

    PubMed Central

    Zhao, Junjing; Chatterjee, Utpal; Ai, Dingfei; Hinks, David G.; Zheng, Hong; Gu, G. D.; Castellan, John-Paul; Rosenkranz, Stephan; Claus, Helmut; Norman, Michael R.; Randeria, Mohit; Campuzano, Juan Carlos

    2013-01-01

    The energy gap for electronic excitations is one of the most important characteristics of the superconducting state, as it directly reflects the pairing of electrons. In the copper–oxide high-temperature superconductors (HTSCs), a strongly anisotropic energy gap, which vanishes along high-symmetry directions, is a clear manifestation of the d-wave symmetry of the pairing. There is, however, a dramatic change in the form of the gap anisotropy with reduced carrier concentration (underdoping). Although the vanishing of the gap along the diagonal to the square Cu–O bond directions is robust, the doping dependence of the large gap along the Cu–O directions suggests that its origin might be different from pairing. It is thus tempting to associate the large gap with a second-order parameter distinct from superconductivity. We use angle-resolved photoemission spectroscopy to show that the two-gap behavior and the destruction of well-defined electronic excitations are not universal features of HTSCs, and depend sensitively on how the underdoped materials are prepared. Depending on cation substitution, underdoped samples either show two-gap behavior or not. In contrast, many other characteristics of HTSCs, such as the dome-like dependence of on doping, long-lived excitations along the diagonals to the Cu–O bonds, and an energy gap at the Brillouin zone boundary that decreases monotonically with doping while persisting above (the pseudogap), are present in all samples, irrespective of whether they exhibit two-gap behavior or not. Our results imply that universal aspects of high- superconductivity are relatively insensitive to differences in the electronic states along the Cu–O bond directions. PMID:24101464

  17. Proteins related to the spindle and checkpoint mitotic emphasize the different pathogenesis of hypoplastic MDS.

    PubMed

    Heredia, Fabiola Fernandes; de Sousa, Juliana Cordeiro; Ribeiro Junior, Howard Lopes; Carvalho, Alex Fiorini; Magalhaes, Silvia Maria Meira; Pinheiro, Ronald Feitosa

    2014-02-01

    Some studies show that alterations in expression of proteins related to mitotic spindle (AURORAS KINASE A and B) and mitotic checkpoint (CDC20 and MAD2L1) are involved in chromosomal instability and tumor progression in various solid and hematologic malignancies. This study aimed to evaluate these genes in MDS patients. The cytogenetics analysis was carried out by G-banding, AURKA and AURKB amplification was performed using FISH, and AURKA, AURKB, CDC20 and MAD2L1 gene expression was performed by qRT-PCR in 61 samples of bone marrow from MDS patients. AURKA gene amplification was observed in 10% of the cases, which also showed higher expression levels than the control group (p=0.038). Patients with normo/hypercellular BM presented significantly higher expression levels than hypocellular BM patients, but normo and hypercellular BM groups did not differ. After logistic regression analysis, our results showed that HIGH expression levels were associated with increased risk of developing normo/hypercellular MDS. It also indicated that age is associated with AURKA, CDC20 and MAD2L1 HIGH expression levels. The distinct expression of hypocellular patients emphasizes the prognostic importance of cellularity to MDS. The amplification/high expression of AURKA suggests that the increased expression of this gene may be related to the pathogenesis of disease. PMID:24314588

  18. High-spectral resolution observations of the 3.29 micron emission feature: Comparison to QCC and PAHs

    NASA Technical Reports Server (NTRS)

    Tokunaga, Alan T.; Sellgren, Kris; Sakata, Akira; Wada, S.; Onaka, Takashi; Nakada, Y.; Nagata, T.

    1989-01-01

    Two of the most promising explanations for the origin of the interstellar emission features observed at 3.29, 3.4, 6.2, 7.7, 8.6, and 11.3 microns are: quenched carbonaceous composite (QCC) and polycyclic aromatic hydrocarbons (PAHs). High resolution spectra are given of the 3.29 micron emission feature which were taken with the Cooled Grating Array Spectrometer at the NASA Infrared Telescope Facility and previously published. These spectra show that the peak wavelength of the 3.29 micron feature is located at 3.295 + or - 0.005 micron and that it is coincident with the peak absorbance of QCC. The peak wavelength of the 3.29 micron feature appears to be the same in all of the sources observed thus far. However, the width of the feature in HD 44179 and Elias 1 is only 0.023 micron, which is smaller than the 0.043 micron width in NGC 7027, IRAS 21282+5050, the Orion nebula, and BD+30 deg 3639. Spectra of NGC 7027, QCC, and PAHs is shown. QCC matches the 3.29 micron interstellar emission feature very closely in the wavelength of the peak, and it produces a single feature. On the other hand, PAHs rarely match the peak of the interstellar emission feature, and characteristically produce multiple features.

  19. Condensin targets and reduces unwound DNA structures associated with transcription in mitotic chromosome condensation

    PubMed Central

    Sutani, Takashi; Sakata, Toyonori; Nakato, Ryuichiro; Masuda, Koji; Ishibashi, Mai; Yamashita, Daisuke; Suzuki, Yutaka; Hirano, Tatsuya; Bando, Masashige; Shirahige, Katsuhiko

    2015-01-01

    Chromosome condensation is a hallmark of mitosis in eukaryotes and is a prerequisite for faithful segregation of genetic material to daughter cells. Here we show that condensin, which is essential for assembling condensed chromosomes, helps to preclude the detrimental effects of gene transcription on mitotic condensation. ChIP-seq profiling reveals that the fission yeast condensin preferentially binds to active protein-coding genes in a transcription-dependent manner during mitosis. Pharmacological and genetic attenuation of transcription largely rescue bulk chromosome segregation defects observed in condensin mutants. We also demonstrate that condensin is associated with and reduces unwound DNA segments generated by transcription, providing a direct link between an in vitro activity of condensin and its in vivo function. The human condensin isoform condensin I also binds to unwound DNA regions at the transcription start sites of active genes, implying that our findings uncover a fundamental feature of condensin complexes. PMID:26204128

  20. Clinicopathologic Features and Clinical Outcomes of Esophageal Gastrointestinal Stromal Tumor

    PubMed Central

    Feng, Fan; Tian, Yangzi; Liu, Zhen; Xu, Guanghui; Liu, Shushang; Guo, Man; Lian, Xiao; Fan, Daiming; Zhang, Hongwei

    2016-01-01

    Abstract Clinicopathologic features and clinical outcomes of gastrointestinal stromal tumors (GISTs) in esophagus are limited, because of the relatively rare incidence of esophageal GISTs. Therefore, the aim of the current study was to investigate the clinicopathologic features and clinical outcomes of esophageal GISTs, and to investigate the potential factors that may predict prognosis. Esophageal GIST cases were obtained from our center and from case reports and clinical studies extracted from MEDLINE. Clinicopathologic features and survivals were analyzed and compared with gastric GISTs from our center. The most common location was lower esophagus (86.84%), followed by middle and upper esophagus (11.40% and 1.76%). The majority of esophageal GISTs were classified as high-risk category (70.83%). Mitotic index was correlated with histologic type, mutational status, and tumor size. The 5-year disease-free survival and disease-specific survival were 65.1% and 65.9%, respectively. Tumor size, mitotic index, and National Institutes of Health risk classification were associated with prognosis of esophageal GISTs. Only tumor size, however, was the independent risk factor for the prognosis of esophageal GISTs. In comparison to gastric GISTs, the distribution of tumor size, histologic type, and National Institutes of Health risk classification were significantly different between esophageal GISTs and gastric GISTs. The disease-free survival and disease-specific survival of esophageal GISTs were significantly lower than that of gastric GISTs. The most common location for esophageal GISTs was lower esophagus, and most of the esophageal GISTs are high-risk category. Tumor size was the independent risk factor for the prognosis of esophageal GISTs. Esophageal GISTs differ significantly from gastric GISTs in respect to clinicopathologic features. The prognosis of esophageal GISTs was worse than that of gastric GISTs. PMID:26765432

  1. Etching of Silicon in HBr Plasmas for High Aspect Ratio Features

    NASA Technical Reports Server (NTRS)

    Hwang, Helen H.; Meyyappan, M.; Mathad, G. S.; Ranade, R.

    2002-01-01

    Etching in semiconductor processing typically involves using halides because of the relatively fast rates. Bromine containing plasmas can generate high aspect ratio trenches, desirable for DRAM and MEMS applications, with relatively straight sidewalk We present scanning electron microscope images for silicon-etched trenches in a HBr plasma. Using a feature profile simulation, we show that the removal yield parameter, or number of neutrals removed per incident ion due to all processes (sputtering, spontaneous desorption, etc.), dictates the profile shape. We find that the profile becomes pinched off when the removal yield is a constant, with a maximum aspect ratio (AR) of about 5 to 1 (depth to height). When the removal yield decreases with increasing ion angle, the etch rate increases at the comers and the trench bottom broadens. The profiles have ARs of over 9:1 for yields that vary with ion angle. To match the experimentally observed etched time of 250 s for an AR of 9:1 with a trench width of 0.135 microns, we find that the neutral flux must be 3.336 x 10(exp 17)sq cm/s.

  2. High resolution seismic reflection profiles of Holocene volcanic and tectonic features, Mono Lake, California

    NASA Astrophysics Data System (ADS)

    Jayko, A. S.; Hart, P. E.; Bursik, M. I.; McClain, J. S.; Moore, J. C.; Boyle, M.; Childs, J. R.; Novick, M.; Hill, D. P.; Mangan, M.; Roeske, S.

    2009-12-01

    The Inyo-Mono Craters of Long Valley and Mono Basin, California are the youngest eruptive vents of the Great Basin, USA and the second youngest in California. They are one of two seismically active volcanic centers with geothermal power production in the Walker Lane, western Great Basin, the other being the Coso Volcanic Field to the south. High resolution seismic reflection data collected from the northern tip of the Mono Craters eruptive centers in Mono Lake delinates two structural zones proximal to the active volcanic centers in Mono Lake. A growth structure drapped by ~30 m or more of bedded sediment shows increasing deformation and offset of clastic deposits on the northwest margin of the basin. Coherent thin-bedded stratigraphic sections with strong reflectors to 30-100m depth are preserved on the western and northern margins of the basin. The southern and southeastern areas of the lake are generally seismically opaque, due to extensive ash and tephra deposits as well as widespread methane. Thin pockets of well-bedded, poorly consolidated sediment of probable Holocene and last glacial age are present within intrabasin depressions providing some local age constraints on surfaces adjacent to volcanic vents and volcanically modified features.

  3. Loop-sequence features and stability determinants in antibody variable domains by high-throughput experiments.

    PubMed

    Chang, Hung-Ju; Jian, Jhih-Wei; Hsu, Hung-Ju; Lee, Yu-Ching; Chen, Hong-Sen; You, Jhong-Jhe; Hou, Shin-Chen; Shao, Chih-Yun; Chen, Yen-Ju; Chiu, Kuo-Ping; Peng, Hung-Pin; Lee, Kuo Hao; Yang, An-Suei

    2014-01-01

    Protein loops are frequently considered as critical determinants in protein structure and function. Recent advances in high-throughput methods for DNA sequencing and thermal stability measurement have enabled effective exploration of sequence-structure-function relationships in local protein regions. Using these data-intensive technologies, we investigated the sequence-structure-function relationships of six complementarity-determining regions (CDRs) and ten non-CDR loops in the variable domains of a model vascular endothelial growth factor (VEGF)-binding single-chain antibody variable fragment (scFv) whose sequence had been optimized via a consensus-sequence approach. The results show that only a handful of residues involving long-range tertiary interactions distant from the antigen-binding site are strongly coupled with antigen binding. This implies that the loops are passive regions in protein folding; the essential sequences of these regions are dictated by conserved tertiary interactions and the consensus local loop-sequence features contribute little to protein stability and function. PMID:24268648

  4. High borides: determining the features and details of lattice dynamics from neutron spectroscopy

    NASA Astrophysics Data System (ADS)

    Alekseev, P. A.

    2015-04-01

    We review wide-ranging research that combines inelastic neutron scattering spectroscopy with phenomenological and ab initio calculations to study the lattice dynamics and specifics of the electron-phonon interaction in three-dimensional boron cluster network systems M B_6 and M B12 ( M= {La}, {Sm}, and {Yb}, {Lu}, {Zr}). A close similarity is found between the atomic vibration spectra of these systems, which is fundamentally due to a strong hierarchy of interatomic interaction in these systems and which manifests itself both in the shape of the low-energy phonon dispersion and in the position of the high-energy edge of the spectrum. Manifestations of strong electron-phonon interactions in the lattice vibration spectra of borides are studied in detail and their relation to the nature and features of the valence-unstable state of rare-earth ions is examined. Resonance nonadiabaticity and magnetovibration interaction effects in spin- and valence-fluctuating systems are given special attention.

  5. Binding of multiple features in memory by high-functioning adults with autism spectrum disorder.

    PubMed

    Bowler, Dermot M; Gaigg, Sebastian B; Gardiner, John M

    2014-09-01

    Diminished episodic memory and diminished use of semantic information to aid recall by individuals with autism spectrum disorder (ASD) are both thought to result from diminished relational binding of elements of complex stimuli. To test this hypothesis, we asked high-functioning adults with ASD and typical comparison participants to study grids in which some cells contained drawings of objects in non-canonical colours. Participants were told at study which features (colour, item, location) would be tested in a later memory test. In a second experiment, participants studied similar grids and were told that they would be tested on object-location or object-colour combinations. Recognition of combinations was significantly diminished in ASD, which survived covarying performance on the Color Trails Test (D'Elia et al. Color trails test. Professional manual. Psychological Assessment Resources, Lutz, 1996), a test of executive difficulties. The findings raise the possibility that medial temporal as well as frontal lobe processes are dysfunctional in ASD. PMID:24696375

  6. Some features of bulk melt-textured high-temperature superconductors subjected to alternating magnetic fields

    NASA Astrophysics Data System (ADS)

    Vanderbemden, P.; Molenberg, I.; Simeonova, P.; Lovchinov, V.

    2014-12-01

    Monolithic, large grain, (RE)Ba2Cu3O7 high-temperature superconductors (where RE denotes a rare-earth ion) are known to be able to trap fields in excess of several teslas and represent thus an extremely promising competing technology for permanent magnet in several applications, e.g. in motors and generators. In any rotating machine, however, the superconducting permanent magnet is subjected to variable (transient, or alternating) parasitic magnetic fields. These magnetic fields interact with the superconductor, which yields a reduction of the remnant magnetization. In the present work we quantify these effects by analysing selected experimental data on bulk melt-textured superconductors subjected to AC fields. Our results indicate that the non-uniformity of superconducting properties in rather large samples might lead to unusual features and need to be taken into account to analyse the experimental data. We also investigate the evolution of the DC remnant magnetization of the bulk sample when it is subjected to a large number of AC magnetic field cycles, and investigate the experimental errors that result from a misorientation of the sample or a mispositioning of the Hall probe. The time-dependence of the remnant magnetization over 100000 cycles of the AC field is shown to display distinct regimes which all differ strongly from the usual decay due to magnetic relaxation.

  7. Human Nek7-interactor RGS2 is required for mitotic spindle organization

    PubMed Central

    de Souza, Edmarcia Elisa; Hehnly, Heidi; Perez, Arina Marina; Meirelles, Gabriela Vaz; Smetana, Juliana Helena Costa; Doxsey, Stephen; Kobarg, Jörg

    2015-01-01

    The mitotic spindle apparatus is composed of microtubule (MT) networks attached to kinetochores organized from 2 centrosomes (a.k.a. spindle poles). In addition to this central spindle apparatus, astral MTs assemble at the mitotic spindle pole and attach to the cell cortex to ensure appropriate spindle orientation. We propose that cell cycle-related kinase, Nek7, and its novel interacting protein RGS2, are involved in mitosis regulation and spindle formation. We found that RGS2 localizes to the mitotic spindle in a Nek7-dependent manner, and along with Nek7 contributes to spindle morphology and mitotic spindle pole integrity. RGS2-depletion leads to a mitotic-delay and severe defects in the chromosomes alignment and congression. Importantly, RGS2 or Nek7 depletion or even overexpression of wild-type or kinase-dead Nek7, reduced γ-tubulin from the mitotic spindle poles. In addition to causing a mitotic delay, RGS2 depletion induced mitotic spindle misorientation coinciding with astral MT-reduction. We propose that these phenotypes directly contribute to a failure in mitotic spindle alignment to the substratum. In conclusion, we suggest a molecular mechanism whereupon Nek7 and RGS2 may act cooperatively to ensure proper mitotic spindle organization. PMID:25664600

  8. Tumor Treating Fields Perturb the Localization of Septins and Cause Aberrant Mitotic Exit

    PubMed Central

    Holtzman, Talia S.; Lee, Sze Xian; Wong, Eric T.; Swanson, Kenneth D.

    2015-01-01

    The anti-tumor effects of chemotherapy and radiation are thought to be mediated by triggering G1/S or G2/M cell cycle checkpoints, while spindle poisons, such as paclitaxel, block metaphase exit by initiating the spindle assembly checkpoint. In contrast, we have found that 150 kilohertz (kHz) alternating electric fields, also known as Tumor Treating Fields (TTFields), perturbed cells at the transition from metaphase to anaphase. Cells exposed to the TTFields during mitosis showed normal progression to this point, but exhibited uncontrolled membrane blebbing that coincided with metaphase exit. The ability of such alternating electric fields to affect cellular physiology is likely to be dependent on their interactions with proteins possessing high dipole moments. The mitotic Septin complex consisting of Septin 2, 6 and 7, possesses a high calculated dipole moment of 2711 Debyes (D) and plays a central role in positioning the cytokinetic cleavage furrow, and governing its contraction during ingression. We showed that during anaphase, TTFields inhibited Septin localization to the anaphase spindle midline and cytokinetic furrow, as well as its association with microtubules during cell attachment and spreading on fibronectin. After aberrant metaphase exit as a consequence of TTFields exposure, cells exhibited aberrant nuclear architecture and signs of cellular stress including an overall decrease in cellular proliferation, followed by apoptosis that was strongly influenced by the p53 mutational status. Thus, TTFields are able to diminish cell proliferation by specifically perturbing key proteins involved in cell division, leading to mitotic catastrophe and subsequent cell death. PMID:26010837

  9. Fission yeast pkl1 is a kinesin-related protein involved in mitotic spindle function.

    PubMed Central

    Pidoux, A L; LeDizet, M; Cande, W Z

    1996-01-01

    We have used anti-peptide antibodies raised against highly conserved regions of the kinesin motor domain to identify kinesin-related proteins in the fission yeast Schizosaccharomyces pombe. Here we report the identification of a new kinesin-related protein, which we have named pkl1. Sequence homology and domain organization place pkl1 in the Kar3/ncd subfamily of kinesin-related proteins. Bacterially expressed pkl1 fusion proteins display microtubule-stimulated ATPase activity, nucleotide-sensitive binding, and bundling of microtubules. Immunofluorescence studies with affinity-purified antibodies indicate that the pkl1 protein localizes to the nucleus and the mitotic spindle. Pkl1 null mutants are viable but have increased sensitivity to microtubule-disrupting drugs. Disruption of pkl1+ suppresses mutations in another kinesin-related protein, cut7, which is known to act in the spindle. Overexpression of pkl1 to very high levels causes a similar phenotype to that seen in cut7 mutants: V-shaped and star-shaped microtubule structures are observed, which we interpret to be spindles with unseparated spindle poles. These observations suggest that pkl1 and cut7 provide opposing forces in the spindle. We propose that pkl1 functions as a microtubule-dependent motor that is involved in microtubule organization in the mitotic spindle. Images PMID:8898367

  10. 212Pb-radioimmunotherapy potentiates paclitaxel-induced cell killing efficacy by perturbing the mitotic spindle checkpoint

    PubMed Central

    Yong, K J; Milenic, D E; Baidoo, K E; Brechbiel, M W

    2013-01-01

    Background: Paclitaxel has recently been reported by this laboratory to potentiate the high-LET radiation therapeutic 212Pb-TCMC-trastuzumab, which targets HER2. To elucidate mechanisms associated with this therapy, targeted α-particle radiation therapeutic 212Pb-TCMC-trastuzumab together with paclitaxel was investigated for the treatment of disseminated peritoneal cancers. Methods: Mice bearing human colon cancer LS-174T intraperitoneal xenografts were pre-treated with paclitaxel, followed by treatment with 212Pb-TCMC-trastuzumab and compared with groups treated with paclitaxel alone, 212Pb-TCMC-HuIgG, 212Pb-TCMC-trastuzumab and 212Pb-TCMC-HuIgG after paclitaxel pre-treatment. Results: 212Pb-TCMC-trastuzumab with paclitaxel given 24 h earlier induced increased mitotic catastrophe and apoptosis. The combined modality of paclitaxel and 212Pb-TCMC-trastuzumab markedly reduced DNA content in the S-phase of the cell cycle with a concomitant increase observed in the G2/M-phase. This treatment regimen also diminished phosphorylation of histone H3, accompanied by an increase in multi-micronuclei, or mitotic catastrophe in nuclear profiles and positively stained γH2AX foci. The data suggests, possible effects on the mitotic spindle checkpoint by the paclitaxel and 212Pb-TCMC-trastuzumab treatment. Consistent with this hypothesis, 212Pb-TCMC-trastuzumab treatment in response to paclitaxel reduced expression and phosphorylation of BubR1, which is likely attributable to disruption of a functional Aurora B, leading to impairment of the mitotic spindle checkpoint. In addition, the reduction of BubR1 expression may be mediated by the association of a repressive transcription factor, E2F4, on the promoter region of BubR1 gene. Conclusion: These findings suggest that the sensitisation to therapy of 212Pb-TCMC-trastuzumab by paclitaxel may be associated with perturbation of the mitotic spindle checkpoint, leading to increased mitotic catastrophe and cell death. PMID:23632482

  11. Microdevice having interior cavity with high aspect ratio surface features and associated methods of manufacture and use

    DOEpatents

    Morales, Alfredo M.

    2002-01-01

    A microdevice having interior cavity with high aspect ratio features and ultrasmooth surfaces, and associated method of manufacture and use is described. An LIGA-produced shaped bit is used to contour polish the surface of a sacrificial mandrel. The contoured sacrificial mandrel is subsequently coated with a structural material and the mandrel removed to produce microdevices having micrometer-sized surface features and sub-micrometer RMS surface roughness.

  12. Excess of miRNA-378a-5p perturbs mitotic fidelity and correlates with breast cancer tumourigenesis in vivo

    PubMed Central

    Winsel, S; Mäki-Jouppila, J; Tambe, M; Aure, M R; Pruikkonen, S; Salmela, A-L; Halonen, T; Leivonen, S-K; Kallio, L; Børresen-Dale, A-L; Kallio, M J

    2014-01-01

    Background: Optimal expression and proper function of key mitotic proteins facilitate control and repair processes that aim to prevent loss or gain of chromosomes, a hallmark of cancer. Altered expression of small regulatory microRNAs is associated with tumourigenesis and metastasis but the impact on mitotic signalling has remained unclear. Methods: Cell-based high-throughput screen identified miR-378a-5p as a mitosis perturbing microRNA. Transient transfections, immunofluorescence, western blotting, time-lapse microscopy, FISH and reporter assays were used to characterise the mitotic anomalies by excess miR-378a-5p. Analysis of microRNA profiles in breast tumours was performed. Results: Overexpression of miR-378a-5p induced numerical chromosome changes in cells and abrogated taxol-induced mitotic block via premature inactivation of the spindle assembly checkpoint. Moreover, excess miR-378a-5p triggered receptor tyrosine kinase–MAP kinase pathway signalling, and was associated with suppression of Aurora B kinase. In breast cancer in vivo, we found that high miR-378a-5p levels correlate with the most aggressive, poorly differentiated forms of cancer. Interpretation: Downregulation of Aurora B by excess miR-378a-5p can explain the observed microtubule drug resistance and increased chromosomal imbalance in the microRNA-overexpressing cells. The results suggest that breast tumours may deploy high miR-378a-5p levels to gain growth advantage and antagonise taxane therapy. PMID:25268374

  13. An improved high order texture features extraction method with application to pathological diagnosis of colon lesions for CT colonography

    NASA Astrophysics Data System (ADS)

    Song, Bowen; Zhang, Guopeng; Lu, Hongbing; Wang, Huafeng; Han, Fangfang; Zhu, Wei; Liang, Zhengrong

    2014-03-01

    Differentiation of colon lesions according to underlying pathology, e.g., neoplastic and non-neoplastic, is of fundamental importance for patient management. Image intensity based textural features have been recognized as a useful biomarker for the differentiation task. In this paper, we introduce high order texture features, beyond the intensity, such as gradient and curvature, for that task. Based on the Haralick texture analysis method, we introduce a virtual pathological method to explore the utility of texture features from high order differentiations, i.e., gradient and curvature, of the image intensity distribution. The texture features were validated on database consisting of 148 colon lesions, of which 35 are non-neoplastic lesions, using the random forest classifier and the merit of area under the curve (AUC) of the receiver operating characteristics. The results show that after applying the high order features, the AUC was improved from 0.8069 to 0.8544 in differentiating non-neoplastic lesion from neoplastic ones, e.g., hyperplastic polyps from tubular adenomas, tubulovillous adenomas and adenocarcinomas. The experimental results demonstrated that texture features from the higher order images can significantly improve the classification accuracy in pathological differentiation of colorectal lesions. The gain in differentiation capability shall increase the potential of computed tomography (CT) colonography for colorectal cancer screening by not only detecting polyps but also classifying them from optimal polyp management for the best outcome in personalized medicine.

  14. High-Resolution Seismic Investigation of a Surface Collapse Feature at Weeks Island Salt Dome, Louisiana

    NASA Astrophysics Data System (ADS)

    Miller, R. D.; Xia, J.; Harding, R. S.; Steeples, D. W.

    2005-05-01

    Seismic imaging techniques delineated the subsurface expression of an active sinkhole above a former salt mine at Weeks Island, Louisiana, which was used at the time by the U.S. Department of Energy's Strategic Petroleum Reserve. (The Weeks Island salt dome is no longer part of the Reserve.) The sinkhole, which at the time of the survey was approximately 12 m wide and 11 m deep, is directly over the edge of the upper storage chamber and approximately 60 m above the top of the salt dome. Surface seismic reflections imaged a dramatic bowl-shaped depression in a 28-m-deep reflector spatially consistent with the sinkhole. Two reflections (28 m and 60 m) on multichannel VSP data represent the only velocity and/or density contrasts detected above the top of the salt dome. The 28-m reflector identified on both VSP and surface seismic reflection data is at a depth consistent with the piezometric surface. Considering the high measured permeability and relative geometric severity of the reflection geometry, it is questionable whether this drape in the 28-m reflection is consistent with the water table. Localized velocity variations could account for some of the apparent geometry. The 60-m salt reflection, evident on VSP, can be interpreted on selected processed surface seismic shot gathers, but is difficult to confidently and consistently identify on stacked sections. The sinkhole lies along a northeast-trending acoustic lineament, possibly related to or associated with salt dissolution. The acoustic expression of the sinkhole suggests a localized, predominantly vertical feature. No evidence was discovered to confidently ascertain the mechanism responsible for exposing the salt to unsaturated meteoric water.

  15. Endoscopic features suggesting gastric cancer in biopsy-proven gastric adenoma with high-grade neoplasia

    PubMed Central

    Kim, Jung Ho; Kim, Yoon Jae; An, Jungsuk; Lee, Jong Joon; Cho, Jae Hee; Kim, Kyoung Oh; Chung, Jun-Won; Kwon, Kwang An; Park, Dong Kyun; Kim, Ju Hyun

    2014-01-01

    AIM: To elucidate the endoscopic features that predict the cancer following endoscopic submucosal dissection (ESD) in patients with high-grade neoplasia (HGN). METHODS: We retrospectively analyzed the medical records of patients who underwent ESD of gastric neoplasms from January 2007 to September 2010. ESD was performed in 555 cases involving 550 patients. A total of 112 lesions from 110 consecutive patients were initially diagnosed as HGN without cancer by forceps biopsy, and later underwent ESD. We classified lesions into two groups according to histologic discrepancies between the biopsy and ESD diagnosis. Gastric adenoma in the final diagnosis by ESD specimens were defined as adenoma group. Lesions with coexisting cancer after ESD were defined as cancer group. RESULTS: The mean age was 65.3 years, and 81 patients were male. There was no significant difference in the age or gender distribution between the adenoma (n = 52) and cancer (n = 60) groups. Thirty-six of these lesions (32.1%) showed histologic concordance between the forceps biopsy and ESD specimens, 16 (14.3%) showed a downgraded histology (low-grade neoplasia), and 60 (53.6%) showed an upgraded histology (cancer). A red color change of the mucosal surface on endoscopy was found in 27/52 (51.9%) of cases in the adenoma group and in 46/60 (76.7%) of cases in the cancer group (P = 0.006). Ulceration of the mucosal surface on endoscopy was found in 5 (9.6%) of 52 lesions in the adenoma group and in 17 (28.3%) of 60 lesions in the cancer group (P = 0.013). In the multivariate analysis, a reddish surface color change and mucosal ulceration were significant predictive factors correlated with cancer after ESD of the HGN by forceps biopsy. CONCLUSION: HGN with a red color change or mucosal ulceration correlated with the presence of gastric cancer. These finding may help to guide the diagnosis and treatment. PMID:25232257

  16. Influence of the circadian rhythm in cell division on radiation-induced mitotic delay in vivo

    SciTech Connect

    Rubin, N.H.

    1982-01-01

    Mitotic delay is described as a classical response to radiation; however, circadian rhythmicity in cell division in vivo has not been considered by many authors. The present study investigated the relation between fluctuations reported as mitotic delay and recovery in vivo and circadian oscillations in mitotic index in mouse corneal epithelium. One aspect involved single doses (approximately 600 rad) given to mice at different circadian stages. The normal circadian rhythm in cell division was never obliterated. Inhibition of mitosis was evident but unpredictable, ranging from 6 to 15 hr after irradiation. Recovery was evident only during the daily increase in mitotic index of controls. The classical interpretation of recovery from mitotic delay may be in an in vitro phenomenon not reflecting in vivo responses, which are apparently strongly circadian stage dependent. The second portion of the study demonstrated a dose-response effect on length of mitotic delay and, to a lesser extent, degree of recovery.

  17. Glycogen-supplemented mitotic cytosol for analyzing Xenopus egg microtubule organization.

    PubMed

    Groen, Aaron C; Ngyuen, Phuong A; Field, Christine M; Ishihara, Keisuke; Mitchison, Timothy J

    2014-01-01

    Undiluted cytoplasmic extract prepared from unfertilized Xenopus laevis eggs by low-speed centrifugation (CSF extracts) is useful for reconstitution of egg microtubule dynamics and meiosis-II spindle organization, but it suffers limitations for biochemical analysis due to abundant particulates. Here, we describe preparation and the use of fully clarified, undiluted mitotic cytosol derived from CSF extract. Addition of glycogen improves the ability of this cytosol to reconstitute microtubule organization, in part through improved energy metabolism. Using fully clarified, glycogen-supplemented mitotic cytosol, we reconstituted (i) stimulation of microtubule polymerization by Ran.GTP (Groen, Coughlin, & Mitchison, 2011; Ohba, Nakamura, Nishitani, & Nishimoto, 1999) and (ii) self-organization of highly regular bipolar arrays of taxol-stabilized microtubules that we termed "pineapples" (Mitchison, Nguyen, Coughlin, & Groen, 2013). Both systems will be useful for biochemical dissection of spindle assembly mechanisms. We also describe reliable small-scale methods for preparing fluorescent antibody probes that can be used for live imaging in egg extract systems as well as standard immunofluorescence. PMID:24630120

  18. Titanium dioxide nanoparticles induce cytotoxicity and reduce mitotic index in human amniotic fluid-derived cells.

    PubMed

    Acar, M S; Bulut, Z B; Ateş, A; Nami, B; Koçak, N; Yıldız, B

    2015-01-01

    Titanium dioxide (TiO2) nanoparticles (NPs) are commonly used materials present in many consumables for which most people are exposed to. The biological hazards of the NPs on human health have been demonstrated previously. In this study, we aimed to assess the cytotoxicity potency of TiO2 NPs on the primary human amniotic fluid cells. The cells derived from amniotic fluid were treated with different dosages of TiO2 NPs for some periods. Cell adhesion status was assessed using a light microscopic observation. Cell proliferation and cell death rates were determined using trypan blue staining and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Also, mitotic index was determined using fluorescence in situ hybridization with chromosome 8 centromer-specific DNA probe. Disrupted cell adhesion, decreased proliferation, and increased mortality rates were detected in the cells that were treated with TiO2 NPs depending on the dosage (p < 0.001). Also, reduced mitotic index was determined in the cells depending on the time and TiO2 dosage when compared with the controls (p < 0.0001). These results showed that TiO2 NPs have high cytotoxicity for amniotic fluid-derived cells. Therefore, different products containing TiO2 NPs should be used with care, especially for pregnant women. PMID:24717318

  19. Active DNA demethylation in post-mitotic neurons: a reason for optimism.

    PubMed

    Gavin, David P; Chase, Kayla A; Sharma, Rajiv P

    2013-12-01

    Over the last several years proteins involved in base excision repair (BER) have been implicated in active DNA demethylation. We review the literature supporting BER as a means of active DNA demethylation, and explain how the various components function and cooperate to remove the potentially most enduring means of epigenetic gene regulation. Recent evidence indicates that the same pathways implicated during periods of widespread DNA demethylation, such as the erasure of methyl marks in the paternal pronucleus soon after fertilization, are operational in post-mitotic neurons. Neuronal functional identities, defined here as the result of a combination of neuronal subtype, location, and synaptic connections are largely maintained through DNA methylation. Chronic mental illnesses, such as schizophrenia, may be the result of both altered neurotransmitter levels and neurons that have assumed dysfunctional neuronal identities. A limitation of most current psychopharmacological agents is their focus on the former, while not addressing the more profound latter pathophysiological process. Previously, it was believed that active DNA demethylation in post-mitotic neurons was rare if not impossible. If this were the case, then reversing the factors that maintain neuronal identity, would be highly unlikely. The emergence of an active DNA demethylation pathway in the brain is a reason for great optimism in psychiatry as it provides a means by which previously pathological neurons may be reprogrammed to serve a more favorable role. Agents targeting epigenetic processes have shown much promise in this regard, and may lead to substantial gains over traditional pharmacological approaches. PMID:23958448

  20. The Mitotic Checkpoint Complex binds a second CDC20 to inhibit active APC/C

    PubMed Central

    Izawa, Daisuke; Pines, Jonathon

    2014-01-01

    The Spindle Assembly Checkpoint (SAC) maintains genomic stability by delaying chromosome segregation until the last chromosome has attached to the mitotic spindle. The SAC prevents the Anaphase Promoting Complex/Cyclosome (APC/C) ubiquitin ligase from recognising Cyclin B and securin by catalysing the incorporation of the APC/C co-activator, CDC20, into a complex called the Mitotic Checkpoint Complex (MCC). The SAC works through unattached kinetochores generating a diffusible ‘wait anaphase’ signal1,2 that inhibits the APC/C in the cytoplasm, but the nature of this signal remains a key unsolved problem. Moreover, the SAC and the APC/C are highly responsive to each other: the APC/C quickly targets Cyclin B and securin once all the chromosomes attach in metaphase, but is rapidly inhibited should kinetochore attachment be perturbed3,4. How this is achieved is also unknown. Here, we show that the MCC can inhibit a second CDC20 that has already bound and activated the APC/C. We show how the MCC inhibits active APC/C and that this is essential for the SAC. Moreover, this mechanism can prevent anaphase in the absence of kinetochore signalling. Thus, we propose that the diffusible ‘wait anaphase’ signal could be the MCC itself, and explain how reactivating the SAC can rapidly inhibit active APC/C. PMID:25383541

  1. Mitotic activity in chondrocyte transplantation from the in vitro phase to the in vivo phase.

    PubMed

    Peretti, G M; Caruso, E M; Papini Zorli, I; Albisetti, W

    2000-01-01

    The transplantation of devitalized allogenic matrices vehiculating autologous chondrocytes, previously isoled and seeded on them could be a solution to the problem of repairing lesions of the joint cartilage. For the matrix/cell "composite" to be "graftable" the cells must continue to duplicate and produce cartilaginous matrix even after transport in vivo. The present study analyzes the mitotic activity of chondrocytes planted on devitalized allogenic cartilage and grafted in living animals. Chondrocytes of joint cartilage of lambs were isolated enzymatically and then seeded in vitro on devitalized allogenic cartilaginous matrices for 3 weeks. At the end of the co-culture period, these matrix/chondrocyte composites were transplanted in subcutaneous pockets of athymic mice. The experimental and control samples were evaluated subsequent to explantation by histological study and incorporation of tritiated thymidine. The results obtained revealed an important decrease in the values for the incorporation of thymidine beginning from experimental time 0 (pre-implant evaluation) up to day 28 after implantation, followed by a mild increase at the experimental time of 42 days. This study demonstrated the tendency of articular chondrocytes cultivated in vitro and subsequently transplanted in vivo on a support of devitalized allogenic cartilaginous matrix to modify mitotic activity from very high values for the first experimental times, typical of the in vitro phases of cellular expansion, to very low values, more similar to the behavior of articular chondrocytes in vivo. PMID:11569091

  2. High resolution as a key feature to perform accurate ELISPOT measurements using Zeiss KS ELISPOT readers.

    PubMed

    Malkusch, Wolf

    2005-01-01

    The enzyme-linked immunospot (ELISPOT) assay was originally developed for the detection of individual antibody secreting B-cells. Since then, the method has been improved, and ELISPOT is used for the determination of the production of tumor necrosis factor (TNF)-alpha, interferon (IFN)-gamma, or various interleukins (IL)-4, IL-5. ELISPOT measurements are performed in 96-well plates with nitrocellulose membranes either visually or by means of image analysis. Image analysis offers various procedures to overcome variable background intensity problems and separate true from false spots. ELISPOT readers offer a complete solution for precise and automatic evaluation of ELISPOT assays. Number, size, and intensity of each single spot can be determined, printed, or saved for further statistical evaluation. Cytokine spots are always round, but because of floating edges with the background, they have a nonsmooth borderline. Resolution is a key feature for a precise detection of ELISPOT. In standard applications shape and edge steepness are essential parameters in addition to size and color for an accurate spot recognition. These parameters need a minimum spot diameter of 6 pixels. Collecting one single image per well with a standard color camera with 750 x 560 pixels will result in a resolution much too low to get all of the spots in a specimen. IFN-gamma spots may have only 25 microm diameters, and TNF-alpha spots just 15 microm. A 750 x 560 pixel image of a 6-mm well has a pixel size of 12 microm, resulting in only 1 or 2 pixel for a spot. Using a precise microscope optic in combination with a high resolution (1300 x 1030 pixel) integrating digital color camera, and at least 2 x 2 images per well will result in a pixel size of 2.5 microm and, as a minimum, 6 pixel diameter per spot. New approaches try to detect two cytokines per cell at the same time (i.e., IFN-gamma and IL-5). Standard staining procedures produce brownish spots (horseradish peroxidase) and blue spots (alkaline phosphatase). Problems may occur with color overlaps from cells producing both cytokines, resulting in violet spots. The latest experiments therefore try to use fluorescence labels as a marker. Fluorescein isothiocyanate results in green spots and Rhodamine in red spots. Cells producing both cytokines appear yellow. These colors can be separated much easier than the violet, red, and blue, especially using a high resolution. PMID:15937349

  3. Enhancement of spontaneous mitotic recombination by the meiotic mutant spo11-1 in Saccharomyces cerevisiae

    SciTech Connect

    Bruschi, C.V.; Esposito, M.S.

    1983-12-01

    Both nonreciprocal and reciprocal mitotic recombination are enhanced by the recessive mutant spo11-1, which was previously shown to affect meiosis by decreasing recombination and increasing nondisjunction. The mitotic effects are not distributed equally in all chromosomal regions. The genotypes of mitotic recombinants in spo11-1/spo11-1 diploid cells provide further evidence that widely spaced chromosomal markers undergo coincident conversion in mitosis.

  4. Spatial regulation of Cdc55-PP2A by Zds1/Zds2 controls mitotic entry and mitotic exit in budding yeast.

    PubMed

    Rossio, Valentina; Yoshida, Satoshi

    2011-05-01

    Budding yeast CDC55 encodes a regulatory B subunit of the PP2A (protein phosphatase 2A), which plays important roles in mitotic entry and mitotic exit. The spatial and temporal regulation of PP2A is poorly understood, although recent studies demonstrated that the conserved proteins Zds1 and Zds2 stoichiometrically bind to Cdc55-PP2A and regulate it in a complex manner. Zds1/Zds2 promote Cdc55-PP2A function for mitotic entry, whereas Zds1/Zds2 inhibit Cdc55-PP2A function during mitotic exit. In this paper, we propose that Zds1/Zds2 primarily control Cdc55 localization. Cortical and cytoplasmic localization of Cdc55 requires Zds1/Zds2, and Cdc55 accumulates in the nucleus in the absence of Zds1/Zds2. By genetically manipulating the nucleocytoplasmic distribution of Cdc55, we showed that Cdc55 promotes mitotic entry when in the cytoplasm. On the other hand, nuclear Cdc55 prevents mitotic exit. Our analysis defines the long-sought molecular function for the zillion different screens family proteins and reveals the importance of the regulation of PP2A localization for proper mitotic progression. PMID:21536748

  5. Spatial regulation of Cdc55–PP2A by Zds1/Zds2 controls mitotic entry and mitotic exit in budding yeast

    PubMed Central

    Rossio, Valentina

    2011-01-01

    Budding yeast CDC55 encodes a regulatory B subunit of the PP2A (protein phosphatase 2A), which plays important roles in mitotic entry and mitotic exit. The spatial and temporal regulation of PP2A is poorly understood, although recent studies demonstrated that the conserved proteins Zds1 and Zds2 stoichiometrically bind to Cdc55–PP2A and regulate it in a complex manner. Zds1/Zds2 promote Cdc55–PP2A function for mitotic entry, whereas Zds1/Zds2 inhibit Cdc55–PP2A function during mitotic exit. In this paper, we propose that Zds1/Zds2 primarily control Cdc55 localization. Cortical and cytoplasmic localization of Cdc55 requires Zds1/Zds2, and Cdc55 accumulates in the nucleus in the absence of Zds1/Zds2. By genetically manipulating the nucleocytoplasmic distribution of Cdc55, we showed that Cdc55 promotes mitotic entry when in the cytoplasm. On the other hand, nuclear Cdc55 prevents mitotic exit. Our analysis defines the long-sought molecular function for the zillion different screens family proteins and reveals the importance of the regulation of PP2A localization for proper mitotic progression. PMID:21536748

  6. A Centromere-Signaling Network Underlies the Coordination among Mitotic Events.

    PubMed

    Trivedi, Prasad; Stukenberg, P Todd

    2016-02-01

    There is increasing evidence that regulators of the spindle checkpoint, kinetochore-microtubule attachments, and sister chromatid cohesion are part of an interconnected mitotic regulatory circuit with two positive feedback loops and the chromosome passenger complex (CPC) at its center. If true, this conceptual breakthrough needs to be integrated into models of mitosis. In this review, we describe this circuit and point out how the double feedback loops could provide insights into the self-organization of some mitotic processes and the autonomy of every chromosome on the mitotic spindle. We also provide working models for how mitotic events may be coordinated by this circuit. PMID:26705896

  7. High albedo dune features suggest past dune migration and possible geochemical cementation of aeolian sediments on Mars

    NASA Astrophysics Data System (ADS)

    Gardin, Emilie; Bourke, Mary C.; Allemand, Pascal; Quantin, Cathy

    2011-04-01

    High albedo features are identified in association with barchan dunes in an equatorial inter-crater dune field on Mars using images from the MRO mission. This paper describes the morphometric properties of these features and their association with the present barchan dune field. We propose that these features are cemented aeolian deposits that form at the foot of the dune avalanche face. A possible terrestrial analog exists at White Sands National Monument, in south-central New Mexico, USA. The presence of these features suggests past episodes of dune migration in inter-crater dunefields and liquid water in the near sub-surface in sufficient quantity to cause the cementation of aeolian dune sediment.

  8. Redundant safety features in a high-channel-count retinal neurostimulator

    PubMed Central

    Kelly, Shawn K.; Ellersick, William F.; Krishnan, Ashwati; Doyle, Patrick; Shire, Douglas B.; Wyatt, John L.; Rizzo, Joseph F.

    2016-01-01

    Safety features embedded in a 256-channel retinal prosthesis integrated circuit are presented. The biology of the retina and the electrochemistry of the electrode-tissue interface demand careful planning and design of the safety features of an implantable retinal stimulation device. We describe the internal limits and communication safety features of our ASIC, but we focus on monitoring and protection circuits for the electrode-tissue interface. Two independent voltage monitoring circuits for each channel measure the electrode polarization voltage at two different times in the biphasic stimulation cycle. The monitors ensure that the charged electrode stays within the electrochemical water window potentials, and that the discharged electrode is within a small window near the counter electrode potential. A switch to connect each electrode to the counter electrode between pulses protects against a wide range of device failures. Additionally, we describe work on an active feedback system to ensure that the electrode voltage is at zero.

  9. Comparison of Aerosol Classification from Airborne High Spectral Resolution Lidar and the CALIPSO Vertical Feature Mask

    NASA Astrophysics Data System (ADS)

    Burton, S. P.; Ferrare, R. A.; Omar, A. H.; Hostetler, C. A.; Hair, J. W.; Rogers, R.; Obland, M. D.; Butler, C. F.; Cook, A. L.; Harper, D. B.

    2012-12-01

    The NASA Langley Research Center (LaRC) airborne High Spectral Resolution Lidar (HSRL-1) on the NASA B200 aircraft has acquired large datasets of aerosol extinction (532nm), backscatter (532 and 1064nm), and depolarization (532 and 1064nm) profiles during 349 science flights in 19 field missions across North America since 2006. The extinction-to-backscatter ratio ("lidar ratio"), aerosol depolarization ratios, and backscatter color ratio measurements from HSRL-1 are scale-invariant parameters that depend on aerosol type but not concentration. These four aerosol intensive parameters are combined to qualitatively classify HSRL aerosol measurements into eight separate composition types. The classification methodology uses models formed from "training cases" with known aerosol type. The remaining measurements are then compared with these models using the Mahalanobis distance. Aerosol products from the CALIPSO satellite include aerosol type information as well, which is used as input to the CALIPSO aerosol retrieval. CALIPSO aerosol types are inferred using a mix of aerosol loading-dependent parameters, estimated aerosol depolarization, and location, altitude, and surface type information. The HSRL instrument flies beneath the CALIPSO satellite orbit track, presenting the opportunity for comparisons between the HSRL aerosol typing and the CALIPSO Vertical Feature Mask Aerosol Subtype product, giving insight into the performance of the CALIPSO aerosol type algorithm. We find that the aerosol classification from the two instruments frequently agree for marine aerosols and pure dust, and somewhat less frequently for pollution and smoke. In addition, the comparison suggests that the CALIPSO polluted dust type is overly inclusive, encompassing cases of dust combined with marine aerosol as well as cases without much evidence of dust. Qualitative classification of aerosol type combined with quantitative profile measurements of aerosol backscatter and extinction has many useful applications. The HSRL products are used to apportion AOT by type and vertical location in the column, and to characterize the frequency of cases where multiple types are present in the column. Resolving scenes with multiple types in the column is not possible with passive imaging radiometer and polarimeter measurements. The HSRL aerosol type also has higher resolution than the CALIPSO layer-wise product and provides insight into the performance of CALIPSO layer separation. Information about the vertical distribution of aerosol types is useful for estimating radiative forcing, understanding aerosol lifetime and transport, and assessing the predictions of transport models. CALIPSO has been a pathfinder, providing the first long-term global data set of aerosol vertical distribution. Based on our results, a future satellite lidar similar to CALIPSO, but with the addition of polarization sensitivity at 1064 nm and the HSRL technique at 532 nm, could provide a significant advance in characterizing the vertical distribution of aerosol.

  10. Ribbon plastic optical fiber linked optical transmitter and receiver modules featuring a high alignment tolerance.

    PubMed

    Lee, Hak-Soon; Park, Jun-Young; Cha, Sang-Mo; Lee, Sang-Shin; Hwang, Gyo-Sun; Son, Yung-Sung

    2011-02-28

    Ribbon plastic optical fiber (POF) linked four-channel optical transmitter (Tx) and receiver (Rx) modules have been proposed and realized featuring an excellent alignment tolerance. The two modules share a common configuration involving an optical sub-assembly (OSA) with vertical cavity surface emitting lasers (VCSELs)/photodetectors (PDs), and their driver ICs, which are integrated onto a single printed circuit board (PCB) substrate. The OSA includes an alignment structure, a beam router and a fiber guide, which were produced by using plastic injection molding. We have accomplished a fully passive alignment between the VCSELs/PDs and the ribbon POF by taking advantage of the alignment structure that serves as a reference during the alignment of the constituent parts of the OSA. The electrical link, which largely determines the operation speed, has been remarkably shortened, due to a direct wire-bonding between the VCSELs/PDs and the driver circuits. The light sources and the detectors can be individually positioned, thereby overcoming the pitch limitations of the ribbon POF, which is made up of perfluorinated graded-index (GI) POF with a 62.5 μm core diameter. The overall alignment tolerance was first assessed by observing the optical coupling efficiency in terms of VCSEL/PD misalignment. The horizontal and vertical 3-dB alignment tolerances were about 20 μm and 150 μm for the Tx and 50 μm and over 200 μm for the Rx, respectively. The VCSEL-to-POF coupling loss for the Tx and the POF-to-PD loss for the Rx were 3.25 dB and 1.35 dB at a wavelength of 850 nm, respectively. Subsequently, a high-speed signal at 3.2 Gb/s was satisfactorily delivered via the Tx and Rx modules over a temperature range of -30 to 70°C with no significant errors; the channel crosstalk was below -30 dB. Finally, the performance of the prepared modules was verified by transmitting a 1080p HDMI video supplied by a Bluelay player to an LCD TV. PMID:21369260

  11. An Educational System to Help Students Assess Website Features and Identify High-Risk Websites

    ERIC Educational Resources Information Center

    Kajiyama, Tomoko; Echizen, Isao

    2015-01-01

    Purpose: The purpose of this paper is to propose an effective educational system to help students assess Web site risk by providing an environment in which students can better understand a Web site's features and determine the risks of accessing the Web site for themselves. Design/methodology/approach: The authors have enhanced a prototype…

  12. Cardinality as a highly descriptive feature in myoelectric pattern recognition for decoding motor volition

    PubMed Central

    Ortiz-Catalan, Max

    2015-01-01

    Accurate descriptors of muscular activity play an important role in clinical practice and rehabilitation research. Such descriptors are features of myoelectric signals extracted from sliding time windows. A wide variety of myoelectric features have been used as inputs to pattern recognition algorithms that aim to decode motor volition. The output of these algorithms can then be used to control limb prostheses, exoskeletons, and rehabilitation therapies. In the present study, cardinality is introduced and compared with traditional time-domain (Hudgins' set) and other recently proposed myoelectric features (for example, rough entropy). Cardinality was found to consistently outperform other features, including those that are more sophisticated and computationally expensive, despite variations in sampling frequency, time window length, contraction dynamics, type, and number of movements (single or simultaneous), and classification algorithms. Provided that the signal resolution is kept between 12 and 14 bits, cardinality improves myoelectric pattern recognition for the prediction of motion volition. This technology is instrumental for the rehabilitation of amputees and patients with motor impairments where myoelectric signals are viable. All code and data used in this work is available online within BioPatRec. PMID:26578873

  13. Analyzing fine-scale wetland composition using high resolution imagery and texture features

    NASA Astrophysics Data System (ADS)

    Szantoi, Zoltan; Escobedo, Francisco; Abd-Elrahman, Amr; Smith, Scot; Pearlstine, Leonard

    2013-08-01

    In order to monitor natural and anthropogenic disturbance effects to wetland ecosystems, it is necessary to employ both accurate and rapid mapping of wet graminoid/sedge communities. Thus, it is desirable to utilize automated classification algorithms so that the monitoring can be done regularly and in an efficient manner. This study developed a classification and accuracy assessment method for wetland mapping of at-risk plant communities in marl prairie and marsh areas of the Everglades National Park. Maximum likelihood (ML) and Support Vector Machine (SVM) classifiers were tested using 30.5 cm aerial imagery, the normalized difference vegetation index (NDVI), first and second order texture features and ancillary data. Additionally, appropriate window sizes for different texture features were estimated using semivariogram analysis. Findings show that the addition of NDVI and texture features increased classification accuracy from 66.2% using the ML classifier (spectral bands only) to 83.71% using the SVM classifier (spectral bands, NDVI and first order texture features).

  14. An Educational System to Help Students Assess Website Features and Identify High-Risk Websites

    ERIC Educational Resources Information Center

    Kajiyama, Tomoko; Echizen, Isao

    2015-01-01

    Purpose: The purpose of this paper is to propose an effective educational system to help students assess Web site risk by providing an environment in which students can better understand a Web site's features and determine the risks of accessing the Web site for themselves. Design/methodology/approach: The authors have enhanced a prototype

  15. Drosophila Wee1 kinase rescues fission yeast from mitotic catastrophe and phosphorylates Drosophila Cdc2 in vitro.

    PubMed Central

    Campbell, S D; Sprenger, F; Edgar, B A; O'Farrell, P H

    1995-01-01

    Cdc2 kinase activity is required for triggering entry into mitosis in all known eukaryotes. Elaborate mechanisms have evolved for regulating Cdc2 activity so that mitosis occurs in a timely manner, when preparations for its execution are complete. In Schizosaccharomyces pombe, Wee1 and a related Mik1 kinase are Cdc2-inhibitory kinases that are required for preventing premature activation of the mitotic program. To identify Cdc2-inhibitory kinases in Drosophila, we screened for cDNA clones that rescue S. pombe wee1- mik1- mutants from lethal mitotic catastrophe. One of the genes identified in this screen, Drosophila wee1 (Dwee1), encodes a new Wee1 homologue. Dwee1 kinase is closely related to human and Xenopus Wee1 homologues, and can inhibit Cdc2 activity by phosphorylating a critical tyrosine residue. Dwee1 mRNA is maternally provided to embryos, and is zygotically expressed during the postblastoderm divisions of embryogenesis. Expression remains high in the proliferating cells of the central nervous system well after cells in the rest of the embryo have ceased dividing. The loss of zygotically expressed Dwee1 does not lead to mitotic catastrophe during postblastoderm cycles 14 to 16. This result may indicate that maternally provided Dwee1 is sufficient for regulating Cdc2 during embryogenesis, or it may reflect the presence of a redundant Cdc2 inhibitory kinase, as in fission yeast. Images PMID:8573790

  16. A high-frequency Doppler feature in the power spectra of simulated GRMHD black hole accretion disks

    SciTech Connect

    Wellons, Sarah; Zhu, Yucong; Narayan, Ramesh; McClintock, Jeffrey E.; Psaltis, Dimitrios

    2014-04-20

    Black hole binaries exhibit a wide range of variability phenomena, from large-scale state changes to broadband noise and quasi-periodic oscillations, but the physical nature of much of this variability is poorly understood. We examine the variability properties of three GRMHD simulations of thin accretion disks around black holes of varying spin, producing light curves and power spectra as would be seen by observers. We find that the simulated power spectra show a broad feature at high frequency, which increases in amplitude with the inclination of the observer. We show that this high-frequency feature is a product of the Doppler effect and that its location is a function of the mass and spin of the black hole. This Doppler feature demonstrates that power spectral properties of the accretion disk can be tied to, and potentially used to determine, physical properties of the black hole.

  17. Endogenous localizome identifies 43 mitotic kinesins in a plant cell

    PubMed Central

    Miki, Tomohiro; Naito, Haruko; Nishina, Momoko; Goshima, Gohta

    2014-01-01

    Kinesins are microtubule (MT)-based motor proteins that have been identified in every eukaryotic species. Intriguingly, land plants have more than 60 kinesins in their genomes, many more than that in yeasts or animals. However, many of these have not yet been characterized, and their cellular functions are unknown. Here, by using endogenous tagging, we comprehensively determined the localization of 72 kinesins during mitosis in the moss Physcomitrella patens. We found that 43 kinesins are localized to mitotic structures such as kinetochores, spindle MTs, or phragmoplasts, which are MT-based structures formed during cytokinesis. Surprisingly, only one of them showed an identical localization pattern to the animal homolog, and many were enriched at unexpected sites. RNA interference and live-cell microscopy revealed postanaphase roles for kinesin-5 in spindle/phragmoplast organization, chromosome segregation, and cytokinesis, which have not been observed in animals. Our study thus provides a list of MT-based motor proteins associated with the cell division machinery in plants. Furthermore, our data challenge the current generalization of determining mitotic kinesin function based solely on studies using yeast and animal cells. PMID:24591632

  18. Mitotic internalization of planar cell polarity proteins preserves tissue polarity.

    PubMed

    Devenport, Danelle; Oristian, Daniel; Heller, Evan; Fuchs, Elaine

    2011-08-01

    Planar cell polarity (PCP) is the collective polarization of cells along the epithelial plane, a process best understood in the terminally differentiated Drosophila wing. Proliferative tissues such as mammalian skin also show PCP, but the mechanisms that preserve tissue polarity during proliferation are not understood. During mitosis, asymmetrically distributed PCP components risk mislocalization or unequal inheritance, which could have profound consequences for the long-range propagation of polarity. Here, we show that when mouse epidermal basal progenitors divide PCP components are selectively internalized into endosomes, which are inherited equally by daughter cells. Following mitosis, PCP proteins are recycled to the cell surface, where asymmetry is re-established by a process reliant on neighbouring PCP. A cytoplasmic dileucine motif governs mitotic internalization of atypical cadherin Celsr1, which recruits Vang2 and Fzd6 to endosomes. Moreover, embryos transgenic for a Celsr1 that cannot mitotically internalize exhibit perturbed hair-follicle angling, a hallmark of defective PCP. This underscores the physiological relevance and importance of this mechanism for regulating polarity during cell division. PMID:21743464

  19. Evidence for mitotic recombination in W sup ei /+ heterozygous mice

    SciTech Connect

    Panthier, J.J.; Condamine, H.; Jacob, F. ); Guenet, J.L. )

    1990-05-01

    A number of alleles at coat color loci of the house mouse give rise to areas of wild-type pigmentation on the coats of otherwise mutant animals. Such unstable alleles include both recessive and dominant mutations. Among the latter are several alleles at the W locus. In this report, phenotypic reversions of the W{sup ei} allele at the W locus were studied. Mice heterozygous in repulsion for both W{sup ei} and buff (bf) (i.e. W{sup ei}+/+bf) were examined for the occurrence of phenotypic reversion events. Buff (bf) is a recessive mutation, which lies 21 cM from W on the telomeric side of chromosome 5 and is responsible for the khaki colored coat of nonagouti buff homozygotes (a/a; bf/bf). Two kinds of fully pigmented reversion spots were recovered on the coats of a/a; W{sup ei}+/+bf mice: either solid black or khaki colored. Furthermore phenotypic reversions of W{sup ei}/+ were enhanced significantly following X-irradiation of 9.25-day-old W{sup ei}/+ embryos (P < 0.04). These observations are consistent with the suggestion of a role for mitotic recombination in the origin of these phenotypic reversions. In addition these results raise the intriguing possibility that some W mutations may enhance mitotic recombination in the house mouse.

  20. Integrin-Linked Kinase Regulates Interphase and Mitotic Microtubule Dynamics

    PubMed Central

    Lim, Simin; Kawamura, Eiko; Fielding, Andrew B.; Maydan, Mykola; Dedhar, Shoukat

    2013-01-01

    Integrin-linked kinase (ILK) localizes to both focal adhesions and centrosomes in distinct multiprotein complexes. Its dual function as a kinase and scaffolding protein has been well characterized at focal adhesions, where it regulates integrin-mediated cell adhesion, spreading, migration and signaling. At the centrosomes, ILK regulates mitotic spindle organization and centrosome clustering. Our previous study showed various spindle defects after ILK knockdown or inhibition that suggested alteration in microtubule dynamics. Since ILK expression is frequently elevated in many cancer types, we investigated the effects of ILK overexpression on microtubule dynamics. We show here that overexpressing ILK in HeLa cells was associated with a shorter duration of mitosis and decreased sensitivity to paclitaxel, a chemotherapeutic agent that suppresses microtubule dynamics. Measurement of interphase microtubule dynamics revealed that ILK overexpression favored microtubule depolymerization, suggesting that microtubule destabilization could be the mechanism behind the decreased sensitivity to paclitaxel, which is known to stabilize microtubules. Conversely, the use of a small molecule inhibitor selective against ILK, QLT-0267, resulted in suppressed microtubule dynamics, demonstrating a new mechanism of action for this compound. We further show that treatment of HeLa cells with QLT-0267 resulted in higher inter-centromere tension in aligned chromosomes during mitosis, slower microtubule regrowth after cold depolymerization and the presence of a more stable population of spindle microtubules. These results demonstrate that ILK regulates microtubule dynamics in both interphase and mitotic cells. PMID:23349730

  1. Mitotic Internalization of Planar Cell Polarity Proteins Preserves Tissue Polarity

    PubMed Central

    Devenport, Danelle; Oristian, Daniel; Heller, Evan; Fuchs, Elaine

    2011-01-01

    Planar cell polarity (PCP) is the collective polarization of cells along the epithelial plane, a process best understood in the terminally differentiated Drosophila wing. Proliferative tissues such as mammalian skin also display PCP, but the mechanisms that preserve tissue polarity during proliferation are not understood. During mitosis, asymmetrically-distributed PCP components risk mislocalisation or unequal inheritance, which could have profound consequences on the long-range propagation of polarity. Here, we show that when mouse epidermal basal progenitors divide, PCP components are selectively internalized into endosomes, which are inherited equally by daughter cells. Following mitosis, PCP proteins are recycled to the cell surface where asymmetry is re-established by a process reliant upon neighbouring PCP. A cytoplasmic dileucine motif governs mitotic internalization of atypical cadherin Celsr1, which recruits Vang2 and Fzd6 to endosomes. Moreover, embryos transgenic for a Celsr1 that cannot mitotically internalize, exhibit perturbed hair follicle angling, a hallmark of defective PCP. This underscores the physiological relevance and importance of this novel mechanism for regulating polarity during cell division. PMID:21743464

  2. Unconventional Functions of Mitotic Kinases in Kidney Tumorigenesis

    PubMed Central

    Hascoet, Pauline; Chesnel, Franck; Le Goff, Cathy; Le Goff, Xavier; Arlot-Bonnemains, Yannick

    2015-01-01

    Human tumors exhibit a variety of genetic alterations, including point mutations, translocations, gene amplifications and deletions, as well as aneuploid chromosome numbers. For carcinomas, aneuploidy is associated with poor patient outcome for a large variety of tumor types, including breast, colon, and renal cell carcinoma. The Renal cell carcinoma (RCC) is a heterogeneous carcinoma consisting of different histologic types. The clear renal cell carcinoma (ccRCC) is the most common subtype and represents 85% of the RCC. Central to the biology of the ccRCC is the loss of function of the Von Hippel–Lindau gene, but is also associated with genetic instability that could be caused by abrogation of the cell cycle mitotic spindle checkpoint and may involve the Aurora kinases, which regulate centrosome maturation. Aneuploidy can also result from the loss of cell–cell adhesion and apical–basal cell polarity that also may be regulated by the mitotic kinases (polo-like kinase 1, casein kinase 2, doublecortin-like kinase 1, and Aurora kinases). In this review, we describe the “non-mitotic” unconventional functions of these kinases in renal tumorigenesis. PMID:26579493

  3. A molecular mechanism of mitotic centrosome assembly in Drosophila

    PubMed Central

    Conduit, Paul T; Richens, Jennifer H; Wainman, Alan; Holder, James; Vicente, Catarina C; Pratt, Metta B; Dix, Carly I; Novak, Zsofia A; Dobbie, Ian M; Schermelleh, Lothar; Raff, Jordan W

    2014-01-01

    Centrosomes comprise a pair of centrioles surrounded by pericentriolar material (PCM). The PCM expands dramatically as cells enter mitosis, but it is unclear how this occurs. In this study, we show that the centriole protein Asl initiates the recruitment of DSpd-2 and Cnn to mother centrioles; both proteins then assemble into co-dependent scaffold-like structures that spread outwards from the mother centriole and recruit most, if not all, other PCM components. In the absence of either DSpd-2 or Cnn, mitotic PCM assembly is diminished; in the absence of both proteins, it appears to be abolished. We show that DSpd-2 helps incorporate Cnn into the PCM and that Cnn then helps maintain DSpd-2 within the PCM, creating a positive feedback loop that promotes robust PCM expansion around the mother centriole during mitosis. These observations suggest a surprisingly simple mechanism of mitotic PCM assembly in flies. DOI: http://dx.doi.org/10.7554/eLife.03399.001 PMID:25149451

  4. Bod1 regulates protein phosphatase 2A at mitotic kinetochores

    PubMed Central

    Porter, Iain M.; Schleicher, Katharina; Porter, Michael; Swedlow, Jason R.

    2013-01-01

    Mitotic entry and progression require the activation of several mitotic kinases and the proper regulation and localization of several phosphatases. The activity and localization of each of these enzymes is tightly controlled through a series of specific activators, inhibitors and regulatory subunits. Two proteins, Ensa and Arpp-19, were recently identified as specific inhibitors of PP2A-B55 and are critical for allowing full activity of Cdk1/cyclin B and entry into mitosis. Here we show that Bod1, a protein required for proper chromosome alignment at mitosis, shares sequence similarity with Ensa and Arpp-19 and specifically inhibits the kinetochore-associated PP2A-B56 holoenzyme. PP2A-B56 regulates the stability of kinetochore-microtubule attachments by dephosphorylating several kinetochore proteins. Loss of Bod1 changes the balance of phosphorylation at kinetochores, causing defects in kinetochore function. Bod1, Ensa and Arpp-19 define a family of specific PP2A inhibitors that regulate specific PP2A holoenzymes at distinct locations and points in the cell cycle. PMID:24157919

  5. Inhibition of mitotic-specific histone phophorylation by sodium arsenite

    SciTech Connect

    Cobo, J.M.; Valdez, J.G.; Gurley, L.R.

    1994-10-01

    Synchronized cultures of Chinese hamster cells (line CHO) were used to measure the effects of 10{mu}M sodium arsenite on histone phosphorylation. This treatment caused cell proliferation to be temporarily arrested, after which the cells spontaneously resumed cell proliferation in a radiomimetric manner. Immediately following treatment, it was found that sodium arsenite affected only mitotic-specific HI and H3 phosphorylations. Neither interphase, nor mitotic, H2A and H4 phosphorylations were affected, nor was interphase HI Phosphorylation affected. The phosphorylation of HI was inhibited only in mitosis, reducing HI phosphorylation to 38.1% of control levels, which was the level of interphase HI phosphorylation. The phosphorylation of both H3 variants was inhibited in mitosis, the less hydrophobic H3 to 19% and the more hydrophobic H3 to 24% of control levels. These results suggest that sodium arsenite may inhibite cell proliferation by interfering with the cyclin B/p34{sup cdc2} histone kinase activity which is thought to play a key role in regulating the cell cycle. It has been proposed by our laboratory that HI and H3 phosphorylations play a role in restructuring interphase chromatin into metaphase chromosomes. Interference of this process by sodium arsenite may lead to structurally damaged chromosomes resulting in the increased cancer risks known to be produced by arsenic exposure from the environment.

  6. Molecular chaperone CCT3 supports proper mitotic progression and cell proliferation in hepatocellular carcinoma cells.

    PubMed

    Zhang, Yuanyuan; Wang, Yuqi; Wei, Youheng; Wu, Jiaxue; Zhang, Pingzhao; Shen, Suqin; Saiyin, Hexige; Wumaier, Reziya; Yang, Xianmei; Wang, Chenji; Yu, Long

    2016-03-01

    CCT3 was one of the subunits of molecular chaperone CCT/TRiC complex, which plays a central role in maintaining cellular proteostasis. We demonstrated that expressions of CCT3 mRNA and protein are highly up-regulated in hepatocellular carcinoma (HCC) tissues, and high level of CCT3 is correlated with poor survival in cancer patients. In HCC cell lines, CCT3 depletion suppresses cell proliferation by inducing mitotic arrest at prometaphase and apoptosis eventually. We also identified CCT3 as a novel regulator of spindle integrity and as a requirement for proper kinetochore-microtubule attachment during mitosis. Moreover, we found that CCT3 depletion sensitizes HCC cells to microtubule destabilizing drug Vincristine. Collectively, our study suggests that CCT3 is indispensible for HCC cell proliferation, and provides a potential drug target for treatment of HCC. PMID:26739059

  7. A Genome Scale Screen for Mutants with Delayed Exit from Mitosis: Ire1-Independent Induction of Autophagy Integrates ER Homeostasis into Mitotic Lifespan

    PubMed Central

    Ghavidel, Ata; Baxi, Kunal; Ignatchenko, Vladimir; Prusinkiewicz, Martin; Arnason, Terra G.; Kislinger, Thomas; Carvalho, Carlos E.; Harkness, Troy A. A.

    2015-01-01

    Proliferating eukaryotic cells undergo a finite number of cell divisions before irreversibly exiting mitosis. Yet pathways that normally limit the number of cell divisions remain poorly characterized. Here we describe a screen of a collection of 3762 single gene mutants in the yeast Saccharomyces cerevisiae, accounting for 2/3 of annotated yeast ORFs, to search for mutants that undergo an atypically high number of cell divisions. Many of the potential longevity genes map to cellular processes not previously implicated in mitotic senescence, suggesting that regulatory mechanisms governing mitotic exit may be broader than currently anticipated. We focused on an ER-Golgi gene cluster isolated in this screen to determine how these ubiquitous organelles integrate into mitotic longevity. We report that a chronic increase in ER protein load signals an expansion in the assembly of autophagosomes in an Ire1-independent manner, accelerates trafficking of high molecular weight protein aggregates from the cytoplasm to the vacuoles, and leads to a profound enhancement of daughter cell production. We demonstrate that this catabolic network is evolutionarily conserved, as it also extends reproductive lifespan in the nematode Caenorhabditis elegans. Our data provide evidence that catabolism of protein aggregates, a natural byproduct of high protein synthesis and turn over in dividing cells, is among the drivers of mitotic longevity in eukaryotes. PMID:26247883

  8. Built-up Areas Extraction in High Resolution SAR Imagery based on the method of Multiple Feature Weighted Fusion

    NASA Astrophysics Data System (ADS)

    Liu, X.; Zhang, J. X.; Zhao, Z.; Ma, A. D.

    2015-06-01

    Synthetic aperture radar in the application of remote sensing technology is becoming more and more widely because of its all-time and all-weather operation, feature extraction research in high resolution SAR image has become a hot topic of concern. In particular, with the continuous improvement of airborne SAR image resolution, image texture information become more abundant. It's of great significance to classification and extraction. In this paper, a novel method for built-up areas extraction using both statistical and structural features is proposed according to the built-up texture features. First of all, statistical texture features and structural features are respectively extracted by classical method of gray level co-occurrence matrix and method of variogram function, and the direction information is considered in this process. Next, feature weights are calculated innovatively according to the Bhattacharyya distance. Then, all features are weighted fusion. At last, the fused image is classified with K-means classification method and the built-up areas are extracted after post classification process. The proposed method has been tested by domestic airborne P band polarization SAR images, at the same time, two groups of experiments based on the method of statistical texture and the method of structural texture were carried out respectively. On the basis of qualitative analysis, quantitative analysis based on the built-up area selected artificially is enforced, in the relatively simple experimentation area, detection rate is more than 90%, in the relatively complex experimentation area, detection rate is also higher than the other two methods. In the study-area, the results show that this method can effectively and accurately extract built-up areas in high resolution airborne SAR imagery.

  9. Change and Dilemma of School Feature Development of Three Junior High Schools in the Remote and Rural Areas of Taiwan

    ERIC Educational Resources Information Center

    Chen, Shan-Hua; Ho, Hsuan-Fu; Yang, Cheng-Cheng

    2012-01-01

    This research is based on qualitative approach and applies in-depth interview with three principals and administrators in three junior high schools located in the remote and rural areas of Taiwan. The aim of this paper was to explore the school feature development process in these three schools. The findings of this study were as follows: most of…

  10. Cascaded ensemble of convolutional neural networks and handcrafted features for mitosis detection

    NASA Astrophysics Data System (ADS)

    Wang, Haibo; Cruz-Roa, Angel; Basavanhally, Ajay; Gilmore, Hannah; Shih, Natalie; Feldman, Mike; Tomaszewski, John; Gonzalez, Fabio; Madabhushi, Anant

    2014-03-01

    Breast cancer (BCa) grading plays an important role in predicting disease aggressiveness and patient outcome. A key component of BCa grade is mitotic count, which involves quantifying the number of cells in the process of dividing (i.e. undergoing mitosis) at a specific point in time. Currently mitosis counting is done manually by a pathologist looking at multiple high power fields on a glass slide under a microscope, an extremely laborious and time consuming process. The development of computerized systems for automated detection of mitotic nuclei, while highly desirable, is confounded by the highly variable shape and appearance of mitoses. Existing methods use either handcrafted features that capture certain morphological, statistical or textural attributes of mitoses or features learned with convolutional neural networks (CNN). While handcrafted features are inspired by the domain and the particular application, the data-driven CNN models tend to be domain agnostic and attempt to learn additional feature bases that cannot be represented through any of the handcrafted features. On the other hand, CNN is computationally more complex and needs a large number of labeled training instances. Since handcrafted features attempt to model domain pertinent attributes and CNN approaches are largely unsupervised feature generation methods, there is an appeal to attempting to combine these two distinct classes of feature generation strategies to create an integrated set of attributes that can potentially outperform either class of feature extraction strategies individually. In this paper, we present a cascaded approach for mitosis detection that intelligently combines a CNN model and handcrafted features (morphology, color and texture features). By employing a light CNN model, the proposed approach is far less demanding computationally, and the cascaded strategy of combining handcrafted features and CNN-derived features enables the possibility of maximizing performance by leveraging the disconnected feature sets. Evaluation on the public ICPR12 mitosis dataset that has 226 mitoses annotated on 35 High Power Fields (HPF, x400 magnification) by several pathologists and 15 testing HPFs yielded an F-measure of 0.7345. Apart from this being the second best performance ever recorded for this MITOS dataset, our approach is faster and requires fewer computing resources compared to extant methods, making this feasible for clinical use.

  11. Feature extraction of bridges for change detection in high resolution SAR data

    NASA Astrophysics Data System (ADS)

    Cadario, Erich; Gross, Hermann; Hammer, Horst; Thiele, Antje; Thoennessen, Ulrich; Schulz, Karsten; Soergel, Uwe; Weydahl, Dan J.

    2008-10-01

    SAR is a remote sensing technique capable to deliver actual data at any time and under bad weather conditions. Before launch of TerraSAR-X, RADARSAT-2, or COSMO-SkyMed, the rather coarse resolution of operational SAR satellite systems allowed an analysis of spaceborne SAR data in case of disaster management only for medium scale products. The new generation of spaceborne SAR satellites permits a more detailed analysis at the object level even for urban areas, which was before restricted to airborne SAR sensors. Change detection in SAR images is an important field of research. In general, the appearance of objects in SAR images strongly depends on the viewing angle and look direction. This makes a comparison of images on a pixel level difficult. The changeover from pixel- to object level leads to the possibility, to look for object-features that are more stable concerning different imaging constellations. Bridges are keyelements of man made infrastructure. In this paper the appearance of bridges in SAR data is analyzed and features are derived that are exploitable for change detection. Here the focus is on analysis at the object level to derive features that are either stable concerning the imaging constellations or that can be predicted based on a given imaging constellation. Thereby, the usage of different sensors will be possible to achieve the goal of real time information. The investigations are supported by simulations, which allow the creation of SAR images for different imaging constellations, bridge materials, and even for situations with destroyed bridges.

  12. Intraventricular pleomorphic xanthoastrocytoma with anaplastic features.

    PubMed

    Fu, Yong-Juan; Miyahara, Hiroaki; Uzuka, Takeo; Natsumeda, Manabu; Okamoto, Kouichirou; Hirose, Takanori; Fujii, Yukihiko; Takahashi, Hitoshi

    2010-08-01

    Pleomorphic xanthoastrocytoma (PXA) is a rare astrocytic tumor that usually occurs in the superficial cerebral hemispheres of children and young adults and has a relatively favorable prognosis. We report an unusual case of supratentorial, intraventricular tumor in a 52-year-old man. The tumor was composed of pleomorphic cells, including giant cells, most of which were multinucleated, and small cells. In addition, frequent xanthic changes in the cytoplasm of the tumor cells, and widespread reticulin deposits and lymphocytic infiltrates in the stroma were characteristic features. Large areas of necrosis were also evident. However, mitotic figures were rare (1-2 mitoses per 10 high-power fields). Many tumor cells were positive for GFAP, and a number were positive for neurofilament protein and synaptophysin, indicating their neuronal differentiation. In addition, occasional tumor cells were positive for CD34. p53 protein was entirely negative in the tumor cells. In diagnosing this tumor histopathologically, differentiation between PXA and giant cell glioblastoma (GCG), a rare variant of glioblastoma, was problematic. However, considering the overall histopathological picture, a final diagnosis of PXA with anaplastic features was made. The present case indicates that PXA can occur as an intraventricular tumor, and suggests that in some instances, it would be very difficult to differentiate PXA and GCG histopathologically. PMID:20051018

  13. DEVELOPING PHYLOGENIES FOR INTEGRATING MITOTIC FUNGI IN THE HYPOCREALES AND DIAPORTHALES

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Recent comprehensive studies of the Hypocreales and Diaporthales using both morphological and molecular characters present the opportunity to integrate the mitotic fungi and to evaluate character evolution of both teleomorphic and anamorphic states. The majority of plant-associated fungi are mitotic...

  14. Development of a computational high-throughput tool for the quantitative examination of dose-dependent histological features.

    PubMed

    Nault, Rance; Colbry, Dirk; Brandenberger, Christina; Harkema, Jack R; Zacharewski, Timothy R

    2015-04-01

    High-resolution digitalizing of histology slides facilitates the development of computational alternatives to manual quantitation of features of interest. We developed a MATLAB-based quantitative histological analysis tool (QuHAnT) for the high-throughput assessment of distinguishable histological features. QuHAnT validation was demonstrated by comparison with manual quantitation using liver sections from mice orally gavaged with sesame oil vehicle or 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD; 0.001-30 μg/kg) every 4 days for 28 days, which elicits hepatic steatosis with mild fibrosis. A quality control module of QuHAnT reduced the number of quantifiable Oil Red O (ORO)-stained images from 3,123 to 2,756. Increased ORO staining was measured at 10 and 30 μg/kg TCDD with a high correlation between manual and computational volume densities (Vv ), although the dynamic range of QuHAnT was 10-fold greater. Additionally, QuHAnT determined the size of each ORO vacuole, which could not be accurately quantitated by visual examination or manual point counting. PicroSirius Red quantitation demonstrated superior collagen deposition detection due to the ability to consider all images within each section. QuHAnT dramatically reduced analysis time and facilitated the comprehensive assessment of features improving accuracy and sensitivity and represents a complementary tool for tissue/cellular features that are difficult and tedious to assess via subjective or semiquantitative methods. PMID:25274660

  15. Development of a Computational High-Throughput Tool for the Quantitative Examination of Dose-Dependent Histological Features

    PubMed Central

    Nault, Rance; Colbry, Dirk; Brandenberger, Christina; Harkema, Jack R.; Zacharewski, Timothy R.

    2015-01-01

    High-resolution digitalizing of histology slides facilitates the development of computational alternatives to manual quantitation of features of interest. We developed a MATLAB-based quantitative histological analysis tool (QuHAnT) for the high-throughput assessment of distinguishable histological features. QuHAnT validation was demonstrated by comparison with manual quantitation using liver sections from mice orally gavaged with sesame oil vehicle or 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD; 0.001–30 µg/kg) every 4 days for 28 days, which elicits hepatic steatosis with mild fibrosis. A quality control module of QuHAnT reduced the number of quantifiable Oil Red O (ORO)-stained images from 3,123 to 2,756. Increased ORO staining was measured at 10 and 30 µg/kg TCDD with a high correlation between manual and computational volume densities (Vv), although the dynamic range of QuHAnT was 10-fold greater. Additionally, QuHAnT determined the size of each ORO vacuole, which could not be accurately quantitated by visual examination or manual point counting. PicroSirius Red quantitation demonstrated superior collagen deposition detection due to the ability to consider all images within each section. QuHAnT dramatically reduced analysis time and facilitated the comprehensive assessment of features improving accuracy and sensitivity and represents a complementary tool for tissue/cellular features that are difficult and tedious to assess via subjective or semiquantitative methods. PMID:25274660

  16. Mitosis detection in breast cancer pathology images by combining handcrafted and convolutional neural network features.

    PubMed

    Wang, Haibo; Cruz-Roa, Angel; Basavanhally, Ajay; Gilmore, Hannah; Shih, Natalie; Feldman, Mike; Tomaszewski, John; Gonzalez, Fabio; Madabhushi, Anant

    2014-10-01

    Breast cancer (BCa) grading plays an important role in predicting disease aggressiveness and patient outcome. A key component of BCa grade is the mitotic count, which involves quantifying the number of cells in the process of dividing (i.e., undergoing mitosis) at a specific point in time. Currently, mitosis counting is done manually by a pathologist looking at multiple high power fields (HPFs) on a glass slide under a microscope, an extremely laborious and time consuming process. The development of computerized systems for automated detection of mitotic nuclei, while highly desirable, is confounded by the highly variable shape and appearance of mitoses. Existing methods use either handcrafted features that capture certain morphological, statistical, or textural attributes of mitoses or features learned with convolutional neural networks (CNN). Although handcrafted features are inspired by the domain and the particular application, the data-driven CNN models tend to be domain agnostic and attempt to learn additional feature bases that cannot be represented through any of the handcrafted features. On the other hand, CNN is computationally more complex and needs a large number of labeled training instances. Since handcrafted features attempt to model domain pertinent attributes and CNN approaches are largely supervised feature generation methods, there is an appeal in attempting to combine these two distinct classes of feature generation strategies to create an integrated set of attributes that can potentially outperform either class of feature extraction strategies individually. We present a cascaded approach for mitosis detection that intelligently combines a CNN model and handcrafted features (morphology, color, and texture features). By employing a light CNN model, the proposed approach is far less demanding computationally, and the cascaded strategy of combining handcrafted features and CNN-derived features enables the possibility of maximizing the performance by leveraging the disconnected feature sets. Evaluation on the public ICPR12 mitosis dataset that has 226 mitoses annotated on 35 HPFs ([Formula: see text] magnification) by several pathologists and 15 testing HPFs yielded an [Formula: see text]-measure of 0.7345. Our approach is accurate, fast, and requires fewer computing resources compared to existent methods, making this feasible for clinical use. PMID:26158062

  17. Mitosis detection in breast cancer pathology images by combining handcrafted and convolutional neural network features

    PubMed Central

    Wang, Haibo; Cruz-Roa, Angel; Basavanhally, Ajay; Gilmore, Hannah; Shih, Natalie; Feldman, Mike; Tomaszewski, John; Gonzalez, Fabio; Madabhushi, Anant

    2014-01-01

    Abstract. Breast cancer (BCa) grading plays an important role in predicting disease aggressiveness and patient outcome. A key component of BCa grade is the mitotic count, which involves quantifying the number of cells in the process of dividing (i.e., undergoing mitosis) at a specific point in time. Currently, mitosis counting is done manually by a pathologist looking at multiple high power fields (HPFs) on a glass slide under a microscope, an extremely laborious and time consuming process. The development of computerized systems for automated detection of mitotic nuclei, while highly desirable, is confounded by the highly variable shape and appearance of mitoses. Existing methods use either handcrafted features that capture certain morphological, statistical, or textural attributes of mitoses or features learned with convolutional neural networks (CNN). Although handcrafted features are inspired by the domain and the particular application, the data-driven CNN models tend to be domain agnostic and attempt to learn additional feature bases that cannot be represented through any of the handcrafted features. On the other hand, CNN is computationally more complex and needs a large number of labeled training instances. Since handcrafted features attempt to model domain pertinent attributes and CNN approaches are largely supervised feature generation methods, there is an appeal in attempting to combine these two distinct classes of feature generation strategies to create an integrated set of attributes that can potentially outperform either class of feature extraction strategies individually. We present a cascaded approach for mitosis detection that intelligently combines a CNN model and handcrafted features (morphology, color, and texture features). By employing a light CNN model, the proposed approach is far less demanding computationally, and the cascaded strategy of combining handcrafted features and CNN-derived features enables the possibility of maximizing the performance by leveraging the disconnected feature sets. Evaluation on the public ICPR12 mitosis dataset that has 226 mitoses annotated on 35 HPFs (400× magnification) by several pathologists and 15 testing HPFs yielded an F-measure of 0.7345. Our approach is accurate, fast, and requires fewer computing resources compared to existent methods, making this feasible for clinical use. PMID:26158062

  18. The Luminous Polycyclic Aromatic Hydrocarbon Emission Features: Applications to High Redshift Galaxies and Active Galactic Nuclei

    NASA Astrophysics Data System (ADS)

    Shipley, Heath V.

    2016-01-01

    For decades, significant work has been applied to calibrating emission from the ultra-violet, nebular emission lines, far-infrared, X-ray and radio as tracers of the star-formation rate (SFR) in distant galaxies. Understanding the exact rate of star-formation and how it evolves with time and galaxy mass has deep implications for how galaxies form. The co-evolution of star-formation and supermassive black hole (SMBH) accretion is one of the key problems in galaxy formation theory. But, many of these SFR indicators are influenced by SMBH accretion in galaxies and result in unreliable SFRs. Utilizing the luminous polycyclic aromatic hydrocarbon (PAH) emission features, I provide a new robust SFR calibration using the luminosity emitted from the PAHs at 6.2μm, 7.7μm and 11.3μm to solve this. The PAH features emit strongly in the mid-infrared (mid-IR; 5-25μm) mitigating dust extinction, containing on average 5-10% of the total IR luminosity in galaxies. I use a sample of 105 star-forming galaxies covering a range of total IR luminosity, LIR = L(8-1000μm) = 109 - 1012 L⊙ and redshift 0 < z < 0.4, with mid-IR spectroscopy from the Spitzer Infrared Spectrograph (IRS), and data covering other SFR indicators (Hα emission and rest-frame 24μm continuum emission). The PAH luminosity correlates linearly with the SFR as measured by the Hα luminosity (corrected for attenuation using the mono-chromatic rest-frame 24μm emission), with a tight scatter of <0.15 dex. The scatter is comparable to that between SFRs derived from the Paα and dust-corrected Hα emission lines. We present a case study in advance of JWST, which will be capable of measuring SFRs (from 8μm rest-frame photometry, i.e. PAHs) in distant galaxies (z ≤ 2) with JWST/MIRI to SFRs as low as ~10 M⊙yr-1, because the PAH features are so bright. We use Spitzer/IRS observations of PAH features in lensed star-forming galaxies at 1 < z < 3 to demonstrate the utility of the PAHs to derive SFRs that agree with those available from Paα. This new SFR indicator will be useful for probing the peak of the SFR density in the universe (1 < z < 3) and for studying the co-evolution of star-formation and supermassive blackhole accretion contemporaneously in a galaxy.

  19. Animal type melanoma: an unusual case with aggressive histological features?

    PubMed

    Russo, Daniela; Vita, Giulia; Ilardi, Gennaro; Siano, Maria; Mascolo, Massimo

    2012-03-15

    Animal-type melanoma (ATM) refers to a well-known but rare, heavily pigmented melanocytic tumor, considered a variant of malignant melanoma, the biological behavior and prognostic significance of which still remain to be completely established. We report a case characterized by proliferation of hyperpigmented epithelioid and spindle cells, focally ulcerating the epidermis. Tumor necrosis, perineural and vascular invasion, as well as a slightly high mitotic index, were also observed. Although there were several features indicating poor prognosis, the lesion was diagnosed as ATM and not as classical melanoma. This almost unique case could help us to confirm the excellent biological behavior of this "uncertain malignant potential tumor", suggesting a biological nature of ATM possibly different from classical melanoma, as reported in other molecular studies. In addition, the first rapid impression of aggressive melanoma could lead the pathologist to render an immediate, incorrect diagnosis. PMID:22309954

  20. The Xenopus TACC homologue, maskin, functions in mitotic spindle assembly.

    PubMed

    O'Brien, Lori L; Albee, Alison J; Liu, Lingling; Tao, Wei; Dobrzyn, Pawel; Lizarraga, Sofia B; Wiese, Christiane

    2005-06-01

    Maskin is the Xenopus homolog of the transforming acidic coiled coil (TACC)-family of microtubule and centrosome-interacting proteins. Members of this family share a approximately 200 amino acid coiled coil motif at their C-termini, but have only limited homology outside of this domain. In all species examined thus far, perturbations of TACC proteins lead to disruptions of cell cycle progression and/or embryonic lethality. In Drosophila, Caenorhabditis elegans, and humans, these disruptions have been attributed to mitotic spindle assembly defects, and the TACC proteins in these organisms are thought to function as structural components of the spindle. In contrast, cell division failure in early Xenopus embryo blastomeres has been attributed to a role of maskin in regulating the translation of, among others, cyclin B1 mRNA. In this study, we show that maskin, like other TACC proteins, plays a direct role in mitotic spindle assembly in Xenopus egg extracts and that this role is independent of cyclin B. Maskin immunodepletion and add-back experiments demonstrate that maskin, or a maskin-associated activity, is required for two distinct steps during spindle assembly in Xenopus egg extracts that can be distinguished by their response to "rescue" experiments. Defects in the "early" step, manifested by greatly reduced aster size during early time points in maskin-depleted extracts, can be rescued by readdition of purified full-length maskin. Moreover, defects in this step can also be rescued by addition of only the TACC-domain of maskin. In contrast, defects in the "late" step during spindle assembly, manifested by abnormal spindles at later time points, cannot be rescued by readdition of maskin. We show that maskin interacts with a number of proteins in egg extracts, including XMAP215, a known modulator of microtubule dynamics, and CPEB, a protein that is involved in translational regulation of important cell cycle regulators. Maskin depletion from egg extracts results in compromised microtubule asters and spindles and the mislocalization of XMAP215, but CPEB localization is unaffected. Together, these data suggest that in addition to its previously reported role as a translational regulator, maskin is also important for mitotic spindle assembly. PMID:15788567

  1. The luminous polycyclic aromatic hydrocarbon emission features: Applications to high redshift galaxies and active galactic nuclei

    NASA Astrophysics Data System (ADS)

    Shipley, Heath Vernon

    The co-evolution of star-formation and supermassive black hole (SMBH) accretion in galaxies is one of the key problems in galaxy formation theory. Understanding the formation of galaxies, and their subsequent evolution, will be coupled to intensive study of the evolution of SMBHs. This thesis focuses on studying diagnostics of star-formation and SMBH accretion to develop tools to study this co-evolution. Chapter 2 consists of using mid-infrared (mid-IR) spectroscopy from the Spitzer Infrared Spectrograph (IRS) to study the nature of star-formation and SMBH accretion. The mid-IR spectra cover wavelengths 5-38mum, spanning the polycyclic aromatic hydrocarbon (PAH) features and important atomic diagnostic lines. We divide our sample into a subsample of galaxies with Spitzer IRAC colors indicative of warm dust heated by an AGN (IRAGN) and those galaxies whose colors indicate star-formation processes (non-IRAGN). In both the IRAGN and star-forming samples, the luminosity in the PAH features correlates strongly with [Ne II]lambda12.8&mum emission line, from which we conclude that the PAH luminosity directly traces the instantaneous star-formation rate (SFR) in both the IRAGN and star-forming galaxies. There is no measurable difference between the PAH luminosity ratios of L11:3/L7:7 and L6:2/L7:7 for the IRAGN and non-IRAGN, suggesting that AGN do not significantly excite or destroy PAH molecules on galaxy-wide scales. In chapter 3, I calibrate the PAH luminosity as a SFR indicator. We provide a new robust SFR calibration using the luminosity emitted from PAH molecules at 6.2mum, 7.7mum and 11.3mum. The PAH features emit strongly in the mid-IR mitigating dust extinction, containing on average 5--10% of the total IR luminosity in galaxies. We use mid-IR spectroscopy from the Spitzer/IRS, and data covering other SFR indicators (Halpha emission and rest-frame 24mum continuum emission). The PAH luminosity correlates linearly with the SFR as measured by the Halpha luminosity (corrected for attenuation using the mono-chromatic rest-frame 24um emission), with a tight scatter of 0.15 dex. The scatter is comparable to that between SFRs derived from the Paalpha and dust-corrected Halpha emission lines, implying the PAH features may be as accurate a SFR indicator as the Hydrogen recombination lines. Because the PAH features are so bright, our PAH SFR calibration enables an efficient way to measure SFRs in distant galaxies with JWST to SFRs as low as ~10 M; yr-1 to z >~ 2. We use Spitzer/IRS observations of PAH features in lensed star-forming galaxies at 1 < z < 3 to demonstrate the utility of the PAHs to derive SFRs as accurate as those available from Paalpha. Chapter 4 is the application of the PAH SFRs for galaxies with AGN to demonstrate the reliability for studies of the co-evolution of star-formation and SMBH accretion. We present a study of the contribution from star-formation in galaxies of varying AGN activity (from pure star-forming galaxies to quasars) as a function of total IR luminosity using a sample of 220 galaxies. We use mid-IR spectroscopy from the Spitzer/IRS and photometry from the MIPS mum, 70mum and 160mum bands with partial coverage of the sample with the Herschel 160mum band for the quasars. The contribution from star-formation to the total IR luminosity implied by the PAH emission decreases with increasing IR luminosity. We find a similar result to previous studies for the correlation between SFR, i.e. PAH luminosity, and AGN luminosity for quasars of LSF [special characters omitted] for the 11.3mum PAH feature only (which has been shown to be the most reliable PAH feature in the vicinity of AGN). This may indicate the PAH luminosity remains a reliable tracer of the SFR for galaxies with strong AGN contributions (i.e. quasars), as we did not subtract off the AGN component before measuring the SFR from the PAH luminosity.

  2. Asynchronous Event-Based Multikernel Algorithm for High-Speed Visual Features Tracking.

    PubMed

    Lagorce, Xavier; Meyer, Cdric; Ieng, Sio-Hoi; Filliat, David; Benosman, Ryad

    2015-08-01

    This paper presents a number of new methods for visual tracking using the output of an event-based asynchronous neuromorphic dynamic vision sensor. It allows the tracking of multiple visual features in real time, achieving an update rate of several hundred kilohertz on a standard desktop PC. The approach has been specially adapted to take advantage of the event-driven properties of these sensors by combining both spatial and temporal correlations of events in an asynchronous iterative framework. Various kernels, such as Gaussian, Gabor, combinations of Gabor functions, and arbitrary user-defined kernels, are used to track features from incoming events. The trackers described in this paper are capable of handling variations in position, scale, and orientation through the use of multiple pools of trackers. This approach avoids the N(2) operations per event associated with conventional kernel-based convolution operations with N N kernels. The tracking performance was evaluated experimentally for each type of kernel in order to demonstrate the robustness of the proposed solution. PMID:25248193

  3. Pancreaticobiliary reflux as a high-risk factor for biliary malignancy: Clinical features and diagnostic advancements

    PubMed Central

    Sugita, Reiji

    2015-01-01

    Pancreaticobiliary junction is composed of complex structure with which biliary duct and pancreatic duct assemble and go out into the ampulla of Vater during duodenum wall surrounding the sphincter of Oddi. Although the sphincter of Oddi functionally prevents the reflux of pancreatic juice, pancreaticobiliary reflux (PBR) occurs when function of the sphincter of Oddi halt. The anatomically abnormal junction is termed pancreaticobiliary maljunction (PBM) and is characterized by pancreatic and bile ducts joining outside of the duodenal wall. PBM is an important anatomical finding because many studies have revealed that biliary malignancies are related due to the carcinogenetic effect of the pancreatic back flow on the biliary mucosa. On the other hand, several studies have been published on the reflux of pancreatic juice into the bile duct without morphological PBM, and the correlation of such cases with biliary diseases, especially biliary malignancies, is drawing considerable attention. Although it has long been possible to diagnose PBM by various imaging modalities, PBR without PBM has remained difficult to assess. Therefore, the pathological features of PBR without PBM have not been yet fully elucidated. Lately, a new method of diagnosing PBR without PBM has appeared, and the features of PBR without PBM should soon be better understood. PMID:26167246

  4. High-Accuracy Detection of Early Parkinson's Disease through Multimodal Features and Machine Learning.

    PubMed

    Prashanth, R; Dutta Roy, Sumantra; Mandal, Pravat K; Ghosh, Shantanu

    2016-06-01

    Early (or preclinical) diagnosis of Parkinson's disease (PD) is crucial for its early management as by the time manifestation of clinical symptoms occur, more than 60% of the dopaminergic neurons have already been lost. It is now established that there exists a premotor stage, before the start of these classic motor symptoms, characterized by a constellation of clinical features, mostly non-motor in nature such as Rapid Eye Movement (REM) sleep Behaviour Disorder (RBD) and olfactory loss. In this paper, we use the non-motor features of RBD and olfactory loss, along with other significant biomarkers such as Cerebrospinal fluid (CSF) measurements and dopaminergic imaging markers from 183 healthy normal and 401 early PD subjects, as obtained from the Parkinson's Progression Markers Initiative (PPMI) database, to classify early PD subjects from normal using Naïve Bayes, Support Vector Machine (SVM), Boosted Trees and Random Forests classifiers. We observe that SVM classifier gave the best performance (96.40% accuracy, 97.03% sensitivity, 95.01% specificity, and 98.88% area under ROC). We infer from the study that a combination of non-motor, CSF and imaging markers may aid in the preclinical diagnosis of PD. PMID:27103193

  5. 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.

  6. Cdc2 phosphorylation of nucleolin demarcates mitotic stages and Alzheimer's disease pathology.

    PubMed

    Dranovsky, A; Vincent, I; Gregori, L; Schwarzman, A; Colflesh, D; Enghild, J; Strittmatter, W; Davies, P; Goldgaber, D

    2001-01-01

    Nucleolin is a major multifunctional nuclear phosphoprotein that is phosphorylated by Cdc2 kinase in mitosis and that participates in a number of cellular processes. The monoclonal antibody TG-3 generated against neurofibrillary tangles (NFT) found in Alzheimer's disease (AD) is highly specific for mitotic cells in culture. We here demonstrate that phosphorylation of nucleolin by Cdc2 kinase generates the TG-3 epitope. The unique pool of TG-3 immunoreactive nucleolin appears abruptly during the prophase. It is associated with chromosomes through the metaphase and it gradually disappears during separation of chromosomes and exit from mitosis. In the brain, nucleolin was localized not only to nuclei but also to neuronal cytoplasm, and it is a marker for early NFT. In patients with AD, Cdc2 phosphorylated nucleolin was present in NFT. These findings suggest that phosphorylation of nucleolin by Cdc2 kinase is a critical event and the point of convergence of two distinct pathways, mitosis and neurodegeneration. PMID:11445251

  7. Mitotic catenation is monitored and resolved by a PKCε-regulated pathway

    PubMed Central

    Brownlow, Nicola; Pike, Tanya; Zicha, Daniel; Collinson, Lucy; Parker, Peter J.

    2014-01-01

    Exit from mitosis is controlled by silencing of the spindle assembly checkpoint (SAC). It is important that preceding exit, all sister chromatid pairs are correctly bioriented, and that residual catenation is resolved, permitting complete sister chromatid separation in the ensuing anaphase. Here we determine that the metaphase response to catenation in mammalian cells operates through PKCε. The PKCε-controlled pathway regulates exit from the SAC only when mitotic cells are challenged by retained catenation and this delayed exit is characterized by BubR1-high and Mad2-low kinetochores. In addition, we show that this pathway is necessary to facilitate resolution of retained catenanes in mitosis. When delayed by catenation in mitosis, inhibition of PKCε results in premature entry into anaphase with PICH-positive strands and chromosome bridging. These findings demonstrate the importance of PKCε-mediated regulation in protection from loss of chromosome integrity in cells failing to resolve catenation in G2. PMID:25483024

  8. Acrylamide effects on kinesin-related proteins of the mitotic/meiotic spindle

    SciTech Connect

    Sickles, Dale W. . E-mail: dsickles@mcg.edu; Sperry, Ann O. . E-mail: sperrya@ecu.edu; Testino, Angie; Friedman, Marvin

    2007-07-01

    The microtubule (MT) motor protein kinesin is a vital component of cells and organs expressing acrylamide (ACR) toxicity. As a mechanism of its potential carcinogenicity, we determined whether kinesins involved in cell division are inhibited by ACR similar to neuronal kinesin [Sickles, D.W., Brady, S.T., Testino, A.R., Friedman, M.A., and Wrenn, R.A. (1996). Direct effect of the neurotoxicant acrylamide on kinesin-based microtubule motility. Journal of Neuroscience Research 46, 7-17.] Kinesin-related genes were isolated from rat testes [Navolanic, P.M., and Sperry, A.O. (2000). Identification of isoforms of a mitotic motor in mammalian spermatogenesis. Biology of Reproduction 62, 1360-1369.], their kinesin-like proteins expressed in bacteria using recombinant DNA techniques and the effects of ACR, glycidamide (GLY) and propionamide (a non-neurotoxic metabolite) on the function of two of the identified kinesin motors were tested. KIFC5A MT bundling activity, required for mitotic spindle formation, was measured in an MT-binding assay. Both ACR and GLY caused a similar concentration-dependent reduction in the binding of MT; concentrations of 100 {mu}M ACR or GLY reduced its activity by 60%. KRP2 MT disassembling activity was assayed using the quantity of tubulin disassembled from taxol-stabilized MT. Both ACR and GLY inhibited KRP2-induced MT disassembly. GLY was substantially more potent; significant reductions of 60% were achieved by 500 {mu}M, a comparable inhibition by ACR required a 5 mM concentration. Propionamide had no significant effect on either kinesin, except KRP2 at 10 mM. This is the first report of ACR inhibition of a mitotic/meiotic motor protein. ACR (or GLY) inhibition of kinesin may be an alternative mechanism to DNA adduction in the production of cell division defects and potential carcinogenicity. We conclude that ACR may act on multiple kinesin family members and produce toxicities in organs highly dependent on microtubule-based functions.

  9. ETV6/RUNX1 abrogates mitotic checkpoint function and targets its key player MAD2L1

    PubMed Central

    Krapf, G; Kaindl, U; Kilbey, A; Fuka, G; Inthal, A; Joas, R; Mann, G; Neil, JC; Haas, OA; Panzer-Grmayer, ER

    2014-01-01

    Approximately 25% of childhood B-cell precursor acute lymphoblastic leukemia have an ETV6/RUNX1 (E/R) gene fusion that results from a t(12;21). This genetic subgroup of leukemia is associated with near-triploidy, near-tetraploidy, and trisomy 21 as rather specific types of secondary changes. Here, we show that, unlike various controls, E/R-expressing Ba/F3 clones acquire a tetraploid karyotype on prolonged culture, corroborating the assumption that E/R may attenuate the mitotic checkpoint (MC). Consistent with this notion, E/R-expressing diploid murine and human cell lines have decreased proportions of cells with 4N DNA content and a lower mitotic index when treated with spindle toxins. Moreover, both RUNX1 and E/R regulate mitotic arrest-deficient 2 L1 (MAD2L1), an essential MC component, by binding to promoter-inherent RUNX1 sites, which results in down-regulation of MAD2L1 mRNA and protein in E/R-expressing cells. Forced expression of E/R also abolishes RUNX1-induced reporter activation, whereas E/R with a mutant DNA-binding site leads to only minor effects. Our data link for the first time E/R, MC, and MAD2L1 and provide new insights into the function of the E/R fusion gene product. Although tetraploidy is an almost exclusive feature of E/R-positive leukemias, its rarity within this particular subgroup implies that further yet unknown factors are required for its manifestation. PMID:20190817

  10. ETV6/RUNX1 abrogates mitotic checkpoint function and targets its key player MAD2L1.

    PubMed

    Krapf, G; Kaindl, U; Kilbey, A; Fuka, G; Inthal, A; Joas, R; Mann, G; Neil, J C; Haas, O A; Panzer-Grmayer, E R

    2010-06-01

    Approximately 25% of childhood B-cell precursor acute lymphoblastic leukemia have an ETV6/RUNX1 (E/R) gene fusion that results from a t(12;21). This genetic subgroup of leukemia is associated with near-triploidy, near-tetraploidy, and trisomy 21 as rather specific types of secondary changes. Here, we show that, unlike various controls, E/R-expressing Ba/F3 clones acquire a tetraploid karyotype on prolonged culture, corroborating the assumption that E/R may attenuate the mitotic checkpoint (MC). Consistent with this notion, E/R-expressing diploid murine and human cell lines have decreased proportions of cells with 4N DNA content and a lower mitotic index when treated with spindle toxins. Moreover, both RUNX1 and E/R regulate mitotic arrest-deficient 2 L1 (MAD2L1), an essential MC component, by binding to promoter-inherent RUNX1 sites, which results in down-regulation of MAD2L1 mRNA and protein in E/R-expressing cells. Forced expression of E/R also abolishes RUNX1-induced reporter activation, whereas E/R with a mutant DNA-binding site leads to only minor effects. Our data link for the first time E/R, MC, and MAD2L1 and provide new insights into the function of the E/R fusion gene product. Although tetraploidy is an almost exclusive feature of E/R-positive leukemias, its rarity within this particular subgroup implies that further yet unknown factors are required for its manifestation. PMID:20190817

  11. Phosphorylation of Xenopus p31comet potentiates mitotic checkpoint exit

    PubMed Central

    Mo, Min; Arnaoutov, Alexei; Dasso, Mary

    2015-01-01

    p31comet plays an important role in spindle assembly checkpoint (SAC) silencing. However, how p31comet's activity is regulated remains unclear. Here we show that the timing of M-phase exit in Xenopus egg extracts (XEEs) depends upon SAC activity, even under conditions that are permissive for spindle assembly. p31comet antagonizes the SAC, promoting XEE progression into anaphase after spindles are fully formed. We further show that mitotic p31comet phosphorylation by Inhibitor of nuclear factor κ-B kinase-β (IKK-β) enhances this role in SAC silencing. Together, our findings implicate IKK-β in the control of anaphase timing in XEE through p31comet activation and SAC downregulation. PMID:25892037

  12. Resurrecting remnants: The lives of post-mitotic midbodies

    PubMed Central

    Chen, Chun-Ting; Ettinger, Andreas W.; Huttner, Wieland B.; Doxsey, Stephen J.

    2014-01-01

    Around a century ago, the midbody was described as a structural assembly within the intercellular bridge during cytokinesis, which served to connect the two future daughter cells. The midbody has become the focus of intense investigation through the identification of a growing number of diverse cellular and molecular pathways that localize to the midbody and contribute to its cytokinetic functions ranging from selective vesicle trafficking, regulated microtubule, actin and ESCRT filament assembly and disassembly, and post-translational modification, such as ubiquitination. More recent studies revealed new and unexpected functions of midbodies that occur in post-mitotic cells. In this article, we provide a historical perspective, discuss exciting new roles for midbodies beyond their cytokinetic function and speculate on their potential contributions to pluripotency. PMID:23245592

  13. Mitotic wavefronts mediated by mechanical signaling in early Drosophila embryos

    NASA Astrophysics Data System (ADS)

    Kang, Louis; Idema, Timon; Liu, Andrea; Lubensky, Tom

    2013-03-01

    Mitosis in the early Drosophila embryo demonstrates spatial and temporal correlations in the form of wavefronts that travel across the embryo in each cell cycle. This coordinated phenomenon requires a signaling mechanism, which we suggest is mechanical in origin. We have constructed a theoretical model that supports nonlinear wavefront propagation in a mechanically-excitable medium. Previously, we have shown that this model captures quantitatively the wavefront speed as it varies with cell cycle number, for reasonable values of the elastic moduli and damping coefficient of the medium. Now we show that our model also captures the displacements of cell nuclei in the embryo in response to the traveling wavefront. This new result further supports that mechanical signaling may play an important role in mediating mitotic wavefronts.

  14. Stochastic Simulation and Graphic Visualization of Mitotic Processes

    PubMed Central

    Gardner, Melissa K; Odde, David J.

    2010-01-01

    Computational modeling can be extremely useful in interpreting experimental results. Here we describe how a relatively sophisticated stochastic model for microtubule dynamic instability in the mitotic spindle can be developed starting with straightforward rules and simple programming code. Once this model is developed, the method for comparing simulation results to experimental data must be carefully considered. The ultimate utility of any computational model relies on its predictive power and the ability to assist in designing new experiments. We describe how “deconstructing” the model through the use of quantitative animations contributes to a better qualitative understanding of model behavior. By extracting key qualitative elements of the model in this fashion, model predictions and new experiments can be more easily extracted from model results. PMID:20096783

  15. Special features of high-speed interaction of supercavitating solids in water

    NASA Astrophysics Data System (ADS)

    Ishchenko, Aleksandr; Akinshin, Ruslan; Afanas'eva, Svetlana; Borisenkov, Igor; Burkin, Viktor; Diachkovskii, Aleksei; Korolkov, Leonid; Moiseev, Dmitrii; Khabibullin, Marat

    2016-01-01

    Special features of material behavior of a supercavitating projectile are investigated at various initial velocities of entering water on the basis of the developed stress-strain state model with possibility of destruction of solids when moving in water and interacting with various underwater barriers with the use of consistent methodological approach of mechanics of continuous media. The calculation-experimental method was used to study the modes of motion of supercavitating projectiles at sub- and supersonic velocities in water medium after acceleration in the barrelled accelerator, as well as their interaction with barriers. Issues of stabilization of the supercavitating projectile on the initial flight path in water were studied. Microphotographs of state of solids made of various materials, before and after interaction with water, at subsonic and supersonic velocities were presented. Supersonic velocity of the supercavitating projectile motion in water of 1590 m/s was recorded.

  16. Edge detection of street trees in high-resolution remote sensing images using spectrum features

    NASA Astrophysics Data System (ADS)

    Zhao, Haohao; Xiao, Pengfeng; Feng, Xuezhi

    2013-10-01

    In this paper a method of Fourier spectrum features based edge detection of urban street trees is described. The QuickBird image was first transformed by 2-D discrete Fourier transform. Then the energy of the component in spatial frequency was calculated. The energy distribution of the angle in max energy was used for further study. Different frequency segments was analyzed, the frequency that can best describe the street tree edge was chosen as the cut-off frequency of the street trees edge. Odd Gabor filter in frequency domain with the cut-off frequency and the max-energy angle was applied for the edge detection. The road center line is extracted by a Gabor filter in frequency domain. Then the edge of the street trees is restricted by the road center line. The edge detection result is analyzed by Canny criteria, and the ?V=1.00, and C=0.89.

  17. Delocalization of the microtubule motor Dynein from mitotic spindles by the human papillomavirus E7 oncoprotein is not sufficient for induction of multipolar mitoses.

    PubMed

    Nguyen, Christine L; McLaughlin-Drubin, Margaret E; Münger, Karl

    2008-11-01

    Dynein is a minus end-directed microtubule motor that transports numerous cargoes throughout the cell. During mitosis, dynein motor activity is necessary for the positioning of spindle microtubules and has also been implicated in inactivating the spindle assembly checkpoint. Mutations in dynein motor and/or accessory proteins are associated with human disease, including cancer, and the delocalization of dynein from mitotic spindles has been correlated with an increased incidence of multipolar spindle formation in some cancer cells that contain supernumerary centrosomes. The high-risk human papillomavirus type 16 (HPV16) E7 oncoprotein induces centrosome overduplication and has been shown to cause multipolar mitotic spindle formation, a diagnostic hallmark of HPV-associated neoplasias. Here, we show that HPV16 E7 expression leads to an increased population of mitotic cells with dynein delocalized from the mitotic spindle. This function maps to sequences of HPV16 E7 that are distinct from the region necessary for centrosome overduplication. However, contrary to previous reports, we provide evidence that dynein delocalization by HPV16 E7 is neither necessary nor sufficient to cause the formation of multipolar mitoses. PMID:18974113

  18. High performance organic integrated device with ultraviolet photodetective and electroluminescent properties consisting of a charge-transfer-featured naphthalimide derivative

    NASA Astrophysics Data System (ADS)

    Wang, Hanyu; Zhou, Jie; Wang, Xu; Lu, Zhiyun; Yu, Junsheng

    2014-08-01

    A high performance organic integrated device (OID) with ultraviolet photodetective and electroluminescent (EL) properties was fabricated by using a charge-transfer-featured naphthalimide derivative of 6-{3,5-bis-[9-(4-t-butylphenyl)-9H-carbazol-3-yl]-phenoxy}-2-(4-t-butylphenyl)-benzo[de]isoquinoline-1,3-dione (CzPhONI) as the active layer. The results showed that the OID had a high detectivity of 1.5 1011 Jones at -3 V under the UV-350 nm illumination with an intensity of 0.6 mW/cm2, and yielded an exciplex EL light emission with a maximum brightness of 1437 cd/m2. Based on the energy band diagram, both the charge transfer feature of CzPhONI and matched energy level alignment were responsible for the dual ultraviolet photodetective and EL functions of OID.

  19. Rabbit System. Low cost, high reliability front end electronics featuring 16 bit dynamic range. [Redundant Analog Bus Based Information Transfer

    SciTech Connect

    Drake, G.; Droege, T.F.; Nelson, C.A. Jr.; Turner, K.J.; Ohska, T.K.

    1985-10-01

    A new crate-based front end system has been built which features low cost, compact packaging, command capability, 16 bit dynamic range digitization, and a high degree of redundancy. The crate can contain a variety of instrumentation modules, and is designed to be situated close to the detector. The system is suitable for readout of a large number of channels via parallel multiprocessor data acquisition.

  20. High-resolution multibeam mapping and submersible surveys of topographic features in the northwestern Gulf of Mexico

    USGS Publications Warehouse

    Hickerson, E.L.; Schmahl, G.P.; Weaver, D.C.; Gardner, J.V.

    2003-01-01

    The Flower Garden Banks National Marine Sanctuary (FGBNMS) and the USGS Pacific Seafloor Mapping Project mapped about 2000 km2 of the northwestern Gulf of Mexico continental shelf during June 2002, using a Kongsberg Simrad EM1000 multibeam echosounder. Mapping focused on select topographic highs thave hae been idetnnfied as biological features warranting protection from oil and gas activities by the Minerals Management Service (MMS). The base maps will be used for all future ROV and submersible missions.

  1. Very high resolution Earth observation features for monitoring plant and animal community structure across multiple spatial scales in protected areas

    NASA Astrophysics Data System (ADS)

    Mairota, Paola; Cafarelli, Barbara; Labadessa, Rocco; Lovergine, Francesco; Tarantino, Cristina; Lucas, Richard M.; Nagendra, Harini; Didham, Raphael K.

    2015-05-01

    Monitoring the status and future trends in biodiversity can be prohibitively expensive using ground-based surveys. Consequently, significant effort is being invested in the use of satellite remote sensing to represent aspects of the proximate mechanisms (e.g., resource availability) that can be related to biodiversity surrogates (BS) such as species community descriptors. We explored the potential of very high resolution (VHR) satellite Earth observation (EO) features as proxies for habitat structural attributes that influence spatial variation in habitat quality and biodiversity change. In a semi-natural grassland mosaic of conservation concern in southern Italy, we employed a hierarchical nested sampling strategy to collect field and VHR-EO data across three spatial extent levels (landscape, patch and plot). Species incidence and abundance data were collected at the plot level for plant, insect and bird functional groups. Spectral and textural VHR-EO image features were derived from a Worldview-2 image. Three window sizes (grains) were tested for analysis and computation of textural features, guided by the perception limits of different organisms. The modelled relationships between VHR-EO features and BS responses differed across scales, suggesting that landscape, patch and plot levels are respectively most appropriate when dealing with birds, plants and insects. This research demonstrates the potential of VHR-EO for biodiversity mapping and habitat modelling, and highlights the importance of identifying the appropriate scale of analysis for specific taxonomic groups of interest. Further, textural features are important in the modelling of functional group-specific indices which represent BS in high conservation value habitat types, and provide a more direct link to species interaction networks and ecosystem functioning, than provided by traditional taxonomic diversity indices.

  2. High-order feature-based mixture models of classification learning predict individual learning curves and enable personalized teaching

    PubMed Central

    Cohen, Yarden; Schneidman, Elad

    2013-01-01

    Pattern classification learning tasks are commonly used to explore learning strategies in human subjects. The universal and individual traits of learning such tasks reflect our cognitive abilities and have been of interest both psychophysically and clinically. From a computational perspective, these tasks are hard, because the number of patterns and rules one could consider even in simple cases is exponentially large. Thus, when we learn to classify we must use simplifying assumptions and generalize. Studies of human behavior in probabilistic learning tasks have focused on rules in which pattern cues are independent, and also described individual behavior in terms of simple, single-cue, feature-based models. Here, we conducted psychophysical experiments in which people learned to classify binary sequences according to deterministic rules of different complexity, including high-order, multicue-dependent rules. We show that human performance on such tasks is very diverse, but that a class of reinforcement learning-like models that use a mixture of features captures individual learning behavior surprisingly well. These models reflect the important role of subjects’ priors, and their reliance on high-order features even when learning a low-order rule. Further, we show that these models predict future individual answers to a high degree of accuracy. We then use these models to build personally optimized teaching sessions and boost learning. PMID:23269833

  3. Clinical Features of Obstructive Sleep Apnea That Determine Its High Prevalence in Resistant Hypertension

    PubMed Central

    Min, Hyun Jin; Cho, Yang-Je; Kim, Chang-Hoon; Kim, Da Hee; Kim, Ha Yan; Choi, Ji In; Lee, Jeung-Gweon

    2015-01-01

    Purpose Resistant hypertension (HTN) occurs in 15-20% of treated hypertensive patients, and 70-80% of resistant hypertensive patients have obstructive sleep apnea (OSA). The characteristics of resistant HTN that predispose patients to OSA have not been reported. Therefore, we aimed to determine the clinical, laboratory, and polysomnographic features of resistant HTN that are significantly associated with OSA. Materials and Methods Hypertensive patients (n=475) who underwent portable polysomnography were enrolled. The patients were categorized into controlled (n=410) and resistant HTN (n=65) groups. The risk factors for the occurrence of OSA in controlled and resistant hypertensive patients were compared, and independent risk factors that are associated with OSA were analyzed. Results Out of 475 patients, 359 (75.6%) were diagnosed with OSA. The prevalence of OSA in resistant HTN was 87.7%, which was significantly higher than that in controlled HTN (73.7%). Age, body mass index, neck circumference, waist circumference, and hip circumference were significantly higher in OSA. However, stepwise multivariate analyses revealed that resistant HTN was not an independent risk factor of OSA. Conclusion The higher prevalence and severity of OSA in resistant HTN may be due to the association of risk factors that are common to both conditions. PMID:26256968

  4. Moving Magnetic Features Around AR 10930 from High-resolution Data Observed by Hinode/SOT

    NASA Astrophysics Data System (ADS)

    Li, Xiaobo; Zhang, Hongqi

    2013-07-01

    We investigate the origin, configuration, and evolution of moving magnetic features (MMFs) in the moat and penumbra regions of NOAA AR 10930 using Hinode/SOT filtergrams and magnetograms. We differentiate MMFs into four types in terms of the location of first appearance and the source of initial flux. The main results are summed up as follows: (1) 50% of the MMFs are produced from or within the penumbra, while 50% are produced within the moat. The MMFs formed in the penumbra normally move outward along radial directions. The MMFs formed in the moat have more dispersed directions of motion. The average speed of most MMFs decreases radially. (2) About 63% of moat fluxes are input by flux emergences. Newly emerged MMFs are normally smaller in size. In their rise phase, they gain flux by adding newly emerging flux or merging other elements, and in the decline phase they lose flux by flux cancellation or fragmentation. The MMFs that are fragments separated from penumbra or other magnetic elements usually have larger flux and longer lifetime. They start their decay process once they are formed. Frequent merging and flux cancellation between MMFs are the dominant factors in MMFs' evolution. (3) Cancellations between opposite-polarity magnetic elements are responsible for most of the low chromospheric bright points. Bipole emergence and MMFs' severance from the penumbra also produce bright points. Elongated or horn-shaped micro-filaments may appear during the separation or cancellation process between magnetic elements.

  5. MOVING MAGNETIC FEATURES AROUND AR 10930 FROM HIGH-RESOLUTION DATA OBSERVED BY HINODE/SOT

    SciTech Connect

    Li Xiaobo; Zhang Hongqi

    2013-07-01

    We investigate the origin, configuration, and evolution of moving magnetic features (MMFs) in the moat and penumbra regions of NOAA AR 10930 using Hinode/SOT filtergrams and magnetograms. We differentiate MMFs into four types in terms of the location of first appearance and the source of initial flux. The main results are summed up as follows: (1) 50% of the MMFs are produced from or within the penumbra, while 50% are produced within the moat. The MMFs formed in the penumbra normally move outward along radial directions. The MMFs formed in the moat have more dispersed directions of motion. The average speed of most MMFs decreases radially. (2) About 63% of moat fluxes are input by flux emergences. Newly emerged MMFs are normally smaller in size. In their rise phase, they gain flux by adding newly emerging flux or merging other elements, and in the decline phase they lose flux by flux cancellation or fragmentation. The MMFs that are fragments separated from penumbra or other magnetic elements usually have larger flux and longer lifetime. They start their decay process once they are formed. Frequent merging and flux cancellation between MMFs are the dominant factors in MMFs' evolution. (3) Cancellations between opposite-polarity magnetic elements are responsible for most of the low chromospheric bright points. Bipole emergence and MMFs' severance from the penumbra also produce bright points. Elongated or horn-shaped micro-filaments may appear during the separation or cancellation process between magnetic elements.

  6. Vibrationally highly excited acetylene as studied by dispersed fluorescence and stimulated emission pumping spectroscopy: Vibrational assignment of the feature states

    NASA Astrophysics Data System (ADS)

    Yamanouchi, Kaoru; Ikeda, Naru; Tsuchiya, Soji; Jonas, David M.; Lundberg, James K.; Adamson, George W.; Field, Robert W.

    1991-11-01

    The dispersed fluorescence (DF) and stimulated emission pumping (SEP) spectra of acetylene originating from single rovibronic levels of the à 1Au state were measured with resolutions of 30 and 0.5 cm-1, respectively, in order to examine the vibrational level structure of the electronic ground X˜ 1Σ+g state. The SEP spectra revealed that the number of vibrational levels under each peak in the DF spectra increases with vibrational energy from a single vibrational level below 8000 cm-1 to as many as ten vibrational levels above 16 500 cm-1. Taking account of the fact that a peak in the DF spectrum in the high energy region is composed of more than one level, a DF peak is called a feature state (or a feature). In the DF spectra from two trans-bending levels (v3=2 and 3) of the à state a total of 140 DF features between 5 700 and 21 200 cm-1 were detected and long progressions in the trans bend (v`4=6 -18) and CC stretch (v■2=0 -6) were identified. Below 14 000 cm-1, 26 out of the 50 observed features were unambiguously assigned to these two modes and represented by a second order anharmonic expansion within the ˜20 cm-1 experimental error. At least three additional trans-bend progressions built on excitation in third vibrational mode were identified. Possible assignments of the third mode to the CH stretch (ν`1) and the cis bend (ν■5) are compared. The Darling-Dennison (DD) resonance between the two degenerate bending modes (trans and cis) was proposed as a mechanism to lend Franck-Condon (FC) intensity to the ν`5 mode. The vibrational analysis of the DF features shows that the DF features correspond to the zero-order FC bright basis states. Each feature represents a group of levels which share the character of a zero-order FC bright level. Above 14 000 cm-1, characteristic groups of DF features with a width of around 300 cm-1 appear in the DF spectra originating from both v3=2 and v'3=3. The relative intensity patterns within each group of features in the two DF spectra are nearly identical. Three anharmonic resonances, including the DD resonance, are proposed as a plausible mechanism which splits a single FC bright state into several DF features. The SEP measurement revealed that a single DF feature splits further into several features with widths around 0.5 cm-1. The characteristic nested level structure identified in the DF and SEP spectra are explained in terms of a stepwise energy flow via a series of anharmonic resonances from the initially excited CC stretch/trans-bend vibrations to the remaining vibrational modes.

  7. A Selective Overview of Variable Selection in High Dimensional Feature Space

    PubMed Central

    Fan, Jianqing

    2010-01-01

    High dimensional statistical problems arise from diverse fields of scientific research and technological development. Variable selection plays a pivotal role in contemporary statistical learning and scientific discoveries. The traditional idea of best subset selection methods, which can be regarded as a specific form of penalized likelihood, is computationally too expensive for many modern statistical applications. Other forms of penalized likelihood methods have been successfully developed over the last decade to cope with high dimensionality. They have been widely applied for simultaneously selecting important variables and estimating their effects in high dimensional statistical inference. In this article, we present a brief account of the recent developments of theory, methods, and implementations for high dimensional variable selection. What limits of the dimensionality such methods can handle, what the role of penalty functions is, and what the statistical properties are rapidly drive the advances of the field. The properties of non-concave penalized likelihood and its roles in high dimensional statistical modeling are emphasized. We also review some recent advances in ultra-high dimensional variable selection, with emphasis on independence screening and two-scale methods. PMID:21572976

  8. Partial inhibition of Cdk1 in G 2 phase overrides the SAC and decouples mitotic events.

    PubMed

    McCloy, Rachael A; Rogers, Samuel; Caldon, C Elizabeth; Lorca, Thierry; Castro, Anna; Burgess, Andrew

    2014-01-01

    Entry and progression through mitosis has traditionally been linked directly to the activity of cyclin-dependent kinase 1 (Cdk1). In this study we utilized low doses of the Cdk1-specific inhibitor, RO3306 from early G 2 phase onwards. Addition of low doses of RO3306 in G 2 phase induced minor chromosome congression and segregation defects. In contrast, mild doses of RO3306 during G 2 phase resulted in cells entering an aberrant mitosis, with cells fragmenting centrosomes and failing to fully disassemble the nuclear envelope. Cells often underwent cytokinesis and metaphase simultaneously, despite the presence of an active spindle assembly checkpoint, which prevented degradation of cyclin B1 and securin, resulting in the random partitioning of whole chromosomes. This highly aberrant mitosis produced a significant increase in the proportion of viable polyploid cells present up to 3 days post-treatment. Furthermore, cells treated with medium doses of RO3306 were only able to reach the threshold of Cdk1 substrate phosphorylation required to initiate nuclear envelope breakdown, but failed to reach the levels of phosphorylation required to correctly complete pro-metaphase. Treatment with low doses of Okadaic acid, which primarily inhibits PP2A, rescued the mitotic defects and increased the number of cells that completed a normal mitosis. This supports the current model that PP2A is the primary phosphatase that counterbalances the activity of Cdk1 during mitosis. Taken together these results show that continuous and subtle disruption of Cdk1 activity from G 2 phase onwards has deleterious consequences on mitotic progression by disrupting the balance between Cdk1 and PP2A. PMID:24626186

  9. Partial inhibition of Cdk1 in G2 phase overrides the SAC and decouples mitotic events

    PubMed Central

    McCloy, Rachael A; Rogers, Samuel; Caldon, C Elizabeth; Lorca, Thierry; Castro, Anna; Burgess, Andrew

    2014-01-01

    Entry and progression through mitosis has traditionally been linked directly to the activity of cyclin-dependent kinase 1 (Cdk1). In this study we utilized low doses of the Cdk1-specific inhibitor, RO3306 from early G2 phase onwards. Addition of low doses of RO3306 in G2 phase induced minor chromosome congression and segregation defects. In contrast, mild doses of RO3306 during G2 phase resulted in cells entering an aberrant mitosis, with cells fragmenting centrosomes and failing to fully disassemble the nuclear envelope. Cells often underwent cytokinesis and metaphase simultaneously, despite the presence of an active spindle assembly checkpoint, which prevented degradation of cyclin B1 and securin, resulting in the random partitioning of whole chromosomes. This highly aberrant mitosis produced a significant increase in the proportion of viable polyploid cells present up to 3 days post-treatment. Furthermore, cells treated with medium doses of RO3306 were only able to reach the threshold of Cdk1 substrate phosphorylation required to initiate nuclear envelope breakdown, but failed to reach the levels of phosphorylation required to correctly complete pro-metaphase. Treatment with low doses of Okadaic acid, which primarily inhibits PP2A, rescued the mitotic defects and increased the number of cells that completed a normal mitosis. This supports the current model that PP2A is the primary phosphatase that counterbalances the activity of Cdk1 during mitosis. Taken together these results show that continuous and subtle disruption of Cdk1 activity from G2 phase onwards has deleterious consequences on mitotic progression by disrupting the balance between Cdk1 and PP2A. PMID:24626186

  10. Applicability of data mining algorithms in the identification of beach features/patterns on high-resolution satellite data

    NASA Astrophysics Data System (ADS)

    Teodoro, Ana C.

    2015-01-01

    The available beach classification algorithms and sediment budget models are mainly based on in situ parameters, usually unavailable for several coastal areas. A morphological analysis using remotely sensed data is a valid alternative. This study focuses on the application of data mining techniques, particularly decision trees (DTs) and artificial neural networks (ANNs) to an IKONOS-2 image in order to identify beach features/patterns in a stretch of the northwest coast of Portugal. Based on knowledge of the coastal features, five classes were defined. In the identification of beach features/patterns, the ANN algorithm presented an overall accuracy of 98.6% and a kappa coefficient of 0.97. The best DTs algorithm (with pruning) presents an overall accuracy of 98.2% and a kappa coefficient of 0.97. The results obtained through the ANN and DTs were in agreement. However, the ANN presented a classification more sensitive to rip currents. The use of ANNs and DTs for beach classification from remotely sensed data resulted in an increased classification accuracy when compared with traditional classification methods. The association of remotely sensed high-spatial resolution data and data mining algorithms is an effective methodology with which to identify beach features/patterns.

  11. Mitotic Asynchrony Induces Transforming Growth Factor-β1 Secretion from Airway Epithelium

    PubMed Central

    Alcala, Sarah E.; Benton, Angela S.; Watson, Alan M.; Kureshi, Suraiya; Reeves, Erica M. K.; Damsker, Jesse; Wang, Zuyi; Nagaraju, Kanneboyina; Anderson, Julia; Williams, Aaron M.; Lee, Amber J. Y.; Hayes, Kathleen; Rose, Mary C.; Hoffman, Eric P.

    2014-01-01

    We recently proposed that mitotic asynchrony in repairing tissue may underlie chronic inflammation and fibrosis, where immune cell infiltration is secondary to proinflammatory cross-talk among asynchronously repairing adjacent tissues. Building on our previous finding that mitotic asynchrony is associated with proinflammatory/fibrotic cytokine secretion (e.g., transforming growth factor [TGF]-β1), here we provide evidence supporting cause-and-effect. Under normal conditions, primary airway epithelial basal cell populations undergo mitosis synchronously and do not secrete proinflammatory or profibrotic cytokines. However, when pairs of nonasthmatic cultures were mitotically synchronized at 12 hours off-set and then combined, the mixed cell populations secreted elevated levels of TGF-β1. This shows that mitotic asynchrony is not only associated with but is also causative of TGF-β1 secretion. The secreted cytokines and other mediators from asthmatic cells were not the cause of asynchronous regeneration; synchronously mitotic nonasthmatic epithelia exposed to conditioned media from asthmatic cells did not show changes in mitotic synchrony. We also tested if resynchronization of regenerating asthmatic airway epithelia reduces TGF-β1 secretion and found that pulse-dosed dexamethasone, simvastatin, and aphidicolin were all effective. We therefore propose a new model for chronic inflammatory and fibrotic conditions where an underlying factor is mitotic asynchrony. PMID:24669775

  12. HIGH-pT Features of z-SCALING at Rhic and Tevatron

    NASA Astrophysics Data System (ADS)

    Tokarev, M. V.; Zborovsky, I.; Dedovich, T. G.

    2008-09-01

    Experimental data on inclusive cross sections of jet, direct photon, and high-pT hadron production in pp/bar pp and AA collisions are analyzed in the framework of z-scaling. The analysis is performed with data obtained at ISR, Sbar ppS, RHIC, and Tevatron. Scaling properties of z-presentation of the inclusive spectra are verified. Physical interpretation of the variable z and the scaling function ψ(z) is discussed. We argue that general principles of self-similarity, locality, and fractality reflect the structure of the colliding objects, interaction of their constituents, and particle formation at small scales. The obtained results suggest that the z-scaling may be used as a tool for searching for new physics phenomena beyond Standard Model in hadron and nucleus collisions at high transverse momentum and high multiplicity at U70, RHIC, Tevatron, and LHC.

  13. The Luminous Polycyclic Aromatic Hydrocarbon Emission Features: Applications to High Redshift Galaxies and Active Galactic Nuclei

    NASA Astrophysics Data System (ADS)

    Shipley, Heath; Papovich, Casey

    2015-08-01

    We provide a new robust star-formation rate (SFR) calibration using the luminosity from polycyclic aromatic hydrogen (PAH) molecules. The PAH features emit strongly in the mid-infrared (mid-IR; 3-19?m), mitigating dust extinction, and they are very luminous, containing 5-10% of the total IR luminosity in galaxies. We derive the calibration of the PAH luminosity as a SFR indicator using a sample of 105 star-forming galaxies covering a range of total IR luminosity, LIR = L(8-1000?m) = 109 - 1012 L? and redshift 0 < z < 0.6. The PAH luminosity correlates linearly with the SFR as measured by the dust-corrected H? luminosity (using the sum of the H? and rest-frame 24?m luminosity from Kennicutt et al. 2009), with tight scatter of ~0.15 dex, comparable to the scatter in the dust-corrected H? SFRs and Pa? SFRs. We show this relation is sensitive to galaxy metallicity, where the PAH luminosity of galaxies with Z < 0.7 Z? departs from the linear SFR relationship but in a behaved manor. We derive for this a correction to galaxies below solar metallicity. As a case study for observations with JWST, we apply the PAH SFR calibration to a sample of lensed galaxies at 1 < z < 3 with Spitzer Infrared Spectrograph (IRS) data, and we demonstrate the utility of PAHs to derive SFRs as accurate as those available from any other indicator. This new SFR indicator will be useful for probing the peak of the SFR density of the universe (1 < z < 3) and for studying the coevolution of star-formation and supermassive blackhole accretion contemporaneously in a galaxy.

  14. Large Erosional Features on the Cascadia Accretionary Wedge Imaged with New High-Resolution Multibeam Bathymetry and Seismic Datasets

    NASA Astrophysics Data System (ADS)

    Beeson, J. W.; Goldfinger, C.

    2013-12-01

    Utilizing new high resolution multibeam bathymetric data along with chirp sub-bottom and multichannel seismic reflection (MCS) data, we identified remarkable erosional features on the toe of the Cascadia accretionary wedge near Willapa Canyon, offshore Washington, USA. Bathymetric data was compiled from the Cascadia Open-Access Seismic Transects (COAST) cruise and from the site survey cruise for the Cascadia Initiative. These features loosely resemble slope failures of the frontal thrust, but can be distinguished from such failures by several key features: They incise the crest of the frontal thrust and encompass the landward limb; They have floors below the level of the abyssal plain, similar to plunge pool morphology; They show no evidence of landslide blocks at the base of the slope indicative of block sliding. The features where likely formed during the latest Pleistocene based on post event deposition, cross-cutting relationships with Juan de Fuca Channel and the Willapa Channel levees and wave field, and post event slip on the frontal thrust of the Cascadia accretionary prism. The Holocene levees of both Willapa Channel and Juan de Fuca Channel overlap these older features, and clearly place an upper bound on the age of the erosional features in the latest Pleistocene. A lower bound is estimated from a sub-bottom profile that images ~30 meters of post scour sediment fill. Using existing literature of Holocene and Pleistocene sedimentation rates we estimate a lower age bound between ~23,000 - 56,000 y.b.p. We also map a fault scarp within the erosional feature, with ~60 m of vertical offset. Using multi-channel seismic reflection profiles from the COAST cruise we interpret this scarp as the surface expression of the landward vergent frontal thrust fault. The apparent short duration of the erosional event along the seaward margin of the accretionary wedge, coupled with the presence of the fresh fault scarp within the erosion zone, are indicative of a dormant feature with significant time required to develop the scarp after cessation of the causative process. Based on morphology, dissimilarity with other submarine features, and available age constraints, we infer that these features were most likely formed during the glacial lake outpouring in the Pacific Northwest known as the Missoula floods which occurred 13,000-19,500 y.b.p. The features themselves bear a strong resemblance to 'coulees' formed during the same glacial events onshore, and the outpourings through Willapa Channel are consistent with previous inferences of the deposition of Missoula Flood deposits in Escanaba Trough. If this timing is correct, the slip rate along the Cascadia frontal thrust can be estimated using fault geometry and scarp height as 2.8 - 4.1 mm/yr.

  15. Binding of Multiple Features in Memory by High-Functioning Adults with Autism Spectrum Disorder

    ERIC Educational Resources Information Center

    Bowler, Dermot M.; Gaigg, Sebastian B.; Gardiner, John M.

    2014-01-01

    Diminished episodic memory and diminished use of semantic information to aid recall by individuals with autism spectrum disorder (ASD) are both thought to result from diminished relational binding of elements of complex stimuli. To test this hypothesis, we asked high-functioning adults with ASD and typical comparison participants to study grids in…

  16. Binding of Multiple Features in Memory by High-Functioning Adults with Autism Spectrum Disorder

    ERIC Educational Resources Information Center

    Bowler, Dermot M.; Gaigg, Sebastian B.; Gardiner, John M.

    2014-01-01

    Diminished episodic memory and diminished use of semantic information to aid recall by individuals with autism spectrum disorder (ASD) are both thought to result from diminished relational binding of elements of complex stimuli. To test this hypothesis, we asked high-functioning adults with ASD and typical comparison participants to study grids in

  17. The Inner Nuclear Membrane Protein Src1 Is Required for Stable Post-Mitotic Progression into G1 in Aspergillus nidulans

    PubMed Central

    Osmani, Aysha H.; Osmani, Stephen A.

    2015-01-01

    How membranes and associated proteins of the nuclear envelope (NE) are assembled specifically and inclusively around segregated genomes during exit from mitosis is incompletely understood. Inner nuclear membrane (INM) proteins play key roles by providing links between DNA and the NE. In this study we have investigated the highly conserved INM protein Src1 in Aspergillus nidulans and have uncovered a novel cell cycle response during post mitotic formation of G1 nuclei. Live cell imaging indicates Src1 could have roles during mitotic exit as it preferentially locates to the NE abscission points during nucleokinesis and to the NE surrounding forming daughter G1 nuclei. Deletion analysis further supported this idea revealing that although Src1 is not required for interphase progression or mitosis it is required for stable post-mitotic G1 nuclear formation. This conclusion is based upon the observation that in the absence of Src1 newly formed G1 nuclei are structurally unstable and immediately undergo architectural modifications typical of mitosis. These changes include NPC modifications that stop nuclear transport as well as disassembly of nucleoli. More intriguingly, the newly generated G1 nuclei then cycle between mitotic- and interphase-like states. The findings indicate that defects in post-mitotic G1 nuclear formation caused by lack of Src1 promote repeated failed attempts to generate stable G1 nuclei. To explain this unexpected phenotype we suggest a type of regulation that promotes repetition of defective cell cycle transitions rather than preventing progression past the defective cell cycle transition. We suggest the term “reboot regulation” to define this mode of cell cycle regulation. The findings are discussed in relationship to recent studies showing the Cdk1 master oscillator can entrain subservient oscillators that when uncoupled cause cell cycle transitions to be repeated. PMID:26147902

  18. Atomic Force Microscopy to Study Mechanics of Living Mitotic Mammalian Cells

    NASA Astrophysics Data System (ADS)

    Toyoda, Yusuke; Stewart, Martin P.; Hyman, Anthony A.; Müller, Daniel J.

    2011-08-01

    While biochemical pathways within mitotic cells have been intensively studied, the mechanics of dividing cells is only poorly understood. In our recent report, an experimental system combining fluorescence and atomic force microscopy was set up to study dynamics of mitotic rounding of mammalian cells. We show that cells have a rounding pressure that increases upon mitotic entry. Using specific inhibitors or perturbations, we revealed biological processes required for force generation that underpin the cell rounding shape change during mitosis. The significance of the finding and an outlook are discussed.

  19. Peculiar Magnetotransport Features of Ultranarrow Graphene Nanoribbons under High Magnetic Field.

    PubMed

    Shen, Haoliang; Cresti, Alessandro; Escoffier, Walter; Shi, Yi; Wang, Xinran; Raquet, Bertrand

    2016-02-23

    Through magnetotransport measurements, we investigate ultrasmooth graphene bilayer nanoribbons obtained by multiwall carbon nanotube unzipping, under a high magnetic field up to 55 T. The high quality of the samples allows us to observe a Hall quantization in ribbons as narrow as 20 nm. The presence, for certain samples, of isolated peaks in the resistance plateau is found to be related to a very moderate long-range disorder, which induces magnetic-field-dependent interedge scattering. Tight-binding numerical simulations of electron transport illustrate and confirm this picture. Our study provides important insights into the quantum Hall effect in quasi-1D systems and indicates possible lines for future investigations of the nonchiral edge states induced by zigzag nanoribbon sections. PMID:26649888

  20. Features of creation of highly accurate models of triumphal pylons for archaeological reconstruction

    NASA Astrophysics Data System (ADS)

    Grishkanich, A. S.; Sidorov, I. S.; Redka, D. N.

    2015-12-01

    Cited a measuring operation for determining the geometric characteristics of objects in space and geodetic survey objects on the ground. In the course of the work, data were obtained on a relative positioning of the pylons in space. There are deviations from verticality. In comparison with traditional surveying this testing method is preferable because it allows you to get in semi-automated mode, the CAD model of the object is high for subsequent analysis that is more economical-ly advantageous.

  1. Epidemiological features of pertussis resurgence based on community populations with high vaccination coverage in China.

    PubMed

    Huang, H; Zhu, T; Gao, C; Gao, Z; Liu, Y; Ding, Y; Sun, J; Guo, L; Liu, P; Chen, D; Wang, L; Wu, S; Zhang, Y

    2015-07-01

    Active symptom surveillance was applied to three selected communities ( 160,147 persons) in Tianjin from 2010 to 2012. We examined 1089 individuals showing pertussis-like symptoms, of which 1022 nasopharyngeal specimens were tested for pertussis by polymerase chain reaction and 802 sera for anti-pertussis toxin antibodies. Of the total cases tested, 113 were confirmed, and their demographic, clinical, and vaccination-related data were collected. The annual incidence was 23.52 cases/100,000 persons among communities, which was 16.22 times that obtained via hospital reports for the same period (P < 0.001). The actual incidence in the 15-69 years age group was most significantly underestimated by hospitals, given that it was 43.08 times that of the reported hospital rate. Among the cases aged <15 years, 84.5% were individuals who had been fully vaccinated. The misdiagnosis rate was as high as 94.69%, and only 5.31% of the confirmed pertussis cases were properly diagnosed as pertussis at their first medical visit. Pertussis incidence in China has been severely underestimated and this was in part due to a high misdiagnosis rate. Adolescents and adults have become new high-risk populations. Future work should focus on reinforcing immunization programmes, especially among adolescents and adults. PMID:25286969

  2. Astro-H: New Spectral Features Seen in High-Resolution X-rays

    NASA Astrophysics Data System (ADS)

    Smith, Randall K.; Odaka, Hirokazu; Astro-H Science Working Group

    2015-01-01

    The Soft X-ray Spectrometer (SXS) microcalorimeter on Astro-H will provide the first high-resolution X-ray spectra of diffuse astrophysical sources. One key new type of science will be charge exchange spectroscopy, wherein highly-ionized metals interact with neutral hydrogen, helium, or other material. This has been detected with modest resolution in comets and planets, and is thought to be the origin of at least some of the 1/4 keV soft X-ray background. We will report on the predicted emission that the Astro-H SXS may detector from all of these sources using the recently released AtomdB Charge Exchange spectral model acx, and comment on possible other sources such as starburst galaxies. The SXS will also observe complex high-resolution spectra from other diffuse sources such as overionized supernova remnants and galaxy clusters. We will discuss these in the context of advanced spectral models using the recently released AtomDB v3.0 data and non-equilibrium models.

  3. Features of >130 Gamma-Ray Bursts at high energy: towards the 2nd Fermi LAT GRB catalog

    NASA Astrophysics Data System (ADS)

    Vianello, Giacomo; Omodei, Nicola; Fermi LAT Collaboration

    2016-01-01

    The high-energy emission from Gamma-Ray Bursts is a formidable probe for extreme physics, calling for highly relativistic sources with very large Lorentz factors. Despite the advancements prompted by observations from the Fermi Large Area Telescope and the Fermi Gamma-Ray Burst Monitor, as well as other observatories, many questions remain open, especially on radiative processes and mechanisms. We present here the most extensive search for GRBs at high energies performed so far, featuring a detection efficiency more than 50% better than previous works, and returning more than 130 detections. With this sample size, much larger than the 35 detections presented in the first Fermi/LAT GRB catalog, we are able to assess the characteristics of the population of GRBs at high energy with unprecedented sensitivity. We will review the preliminary results of this work, as well as their interpretation.

  4. Brachytelephalangy with sparing of the fifth distal phalanx: a feature highly suggestive of Keutel syndrome.

    PubMed

    Miller, Stephen F

    2003-03-01

    Keutel syndrome (KS) is a rare, autosomal recessive condition characterized by diffuse cartilaginous calcification, nasal hypoplasia, brachytelephalangy, and peripheral pulmonary stenosis. A review of the literature produced only 15 reported patients, of whom plain radiographs of the hand or a detailed report are available for review in ten. A distinctive pattern of broadening and shortening of the first through fourth distal phalanges, with sparing of the fifth distal phalanx, is seen in seven of these patients. Two additional patients with Keutel syndrome and this identical finding are presented. I suggest that this pattern of brachytelephalangy is sensitive and highly suggestive of the diagnosis of Keutel syndrome. PMID:12612818

  5. Very compact, high-stability electrostatic actuator featuring contact-free self-limiting displacement

    DOEpatents

    Rodgers, M. Steven; Miller, Samuel L.

    2003-01-01

    A compact electrostatic actuator is disclosed for microelectromechanical (MEM) applications. The actuator utilizes stationary and moveable electrodes, with the stationary electrodes being formed on a substrate and the moveable electrodes being supported above the substrate on a frame. The frame provides a rigid structure which allows the electrostatic actuator to be operated at high voltages (up to 190 Volts) to provide a relatively large actuation force compared to conventional electrostatic comb actuators which are much larger in size. For operation at its maximum displacement, the electrostatic actuator is relatively insensitive to the exact value of the applied voltage and provides a self-limiting displacement.

  6. Proposed criteria for recognizing intrastratal deformation features in marine high resolution seismic reflection profiles

    USGS Publications Warehouse

    O'Leary, D. W.; Laine, E.

    1996-01-01

    Intrastratal deformation of marine strata is ordinarily recorded in high-resolution seismic reflection profiles as acoustically transparent or "chaotic" intervals marked by hyperbolic echoes. Intrastratal deformation is easily confused with buried slump or slide deposits formed initially at the sea floor. Correct identification of intrastratal deformation depends on the presence of a warped continuously reflective layer overlying a chaotic/transparent layer. Decollement is the key criterion for identification in seismic reflection profiles. Other criteria include intrusive structures or faults rooted in a chaotic/transparent layer and thickening and thinning of a chaotic/transparent layer with transitions to reflective intervals.

  7. Paclitaxel sensitivity of breast cancer cells requires efficient mitotic arrest and disruption of Bcl-xL/Bak interaction.

    PubMed

    Flores, M Luz; Castilla, Carolina; Ávila, Rainiero; Ruiz-Borrego, Manuel; Sáez, Carmen; Japón, Miguel A

    2012-06-01

    Taxanes are being used for the treatment of breast cancer. However, cancer cells frequently develop resistance to these drugs with the subsequent recurrence of the tumor. MDA-MB-231 and T-47D breast cancer cell lines were used to assess the effect of paclitaxel treatment on apoptosis and cell cycle, the possible mechanisms of paclitaxel resistance as well as the enhancement of paclitaxel-induced apoptosis based on its combination with phenylethyl isothiocyanate (PEITC). T-47D cells undergo apoptosis in response to paclitaxel treatment. The induction of apoptosis was associated with a robust mitotic arrest and the disruption of Bcl-xL/Bak interaction. By contrary, MDA-MB-231 cells were insensitive to paclitaxel-induced apoptosis and this was associated with a high percentage of cells that slip out of paclitaxel-imposed mitotic arrest and also with the maintenance of Bcl-xL/Bak interaction. The sequential treatment of MDA-MB-231 cells with PEITC followed by paclitaxel inhibited the slippage induced by paclitaxel and increased the apoptosis induction achieved with any of the drugs alone. In breast cancer tissues, high Bcl-xL expression was correlated with a shorter time of disease-free survival in patients treated with a chemotherapeutic regimen that contains paclitaxel, in a statistically significant way. Thus, resistance to paclitaxel in MDA-MB-231 cells is related to the inability to disrupt the Bcl-xL/Bak interaction and increased slippage. In this context, the combination of a drug that induces a strong mitotic arrest, such as paclitaxel, with another that inhibits slippage, such as PEITC, translates into increased apoptotic induction. PMID:22076480

  8. Understanding the structural features of high-amylose maize starch through hydrothermal treatment.

    PubMed

    Yang, Jianing; Xie, Fengwei; Wen, Wenqiang; Chen, Ling; Shang, Xiaoqin; Liu, Peng

    2016-03-01

    In this study, high-amylose starches were hydrothermally-treated and the structural changes were monitored with time (up to 12h) using scanning electron microscopy (SEM), confocal laser scanning microscopy (CLSM), small-angle X-ray scattering (SAXS), X-ray diffraction (XRD), and differential scanning calorimetry (DSC). When high-amylose starches were treated in boiling water, half-shell-like granules were observed by SEM, which could be due to the first hydrolysis of the granule inner region (CLSM). This initial hydrolysis could also immediately (0.5h) disrupt the semi-crystalline lamellar regularity (SAXS) and dramatically reduce the crystallinity (XRD); but with prolonged time of hydrothermal treatment (≥2h), might allow the perfection or formation of amylose single helices, resulting in slightly increased crystallinity (XRD and DSC). These results show that the inner region of granules is composed of mainly loosely-packed amylopectin growth rings with semi-crystalline lamellae, which are vulnerable under gelatinization or hydrolysis. In contrast, the periphery is demonstrated to be more compact, possibly composed of amylose and amylopectin helices intertwined with amylose molecules, which require greater energy input (higher temperature) for disintegration. PMID:26708428

  9. Unique features of high-density lipoproteins in the Japanese: in population and in genetic factors.

    PubMed

    Yokoyama, Shinji

    2015-04-01

    Despite its gradual increase in the past several decades, the prevalence of atherosclerotic vascular disease is low in Japan. This is largely attributed to difference in lifestyle, especially food and dietary habits, and it may be reflected in certain clinical parameters. Plasma high-density lipoprotein (HDL) levels, a strong counter risk for atherosclerosis, are indeed high among the Japanese. Accordingly, lower HDL seems to contribute more to the development of coronary heart disease (CHD) than an increase in non-HDL lipoproteins at a population level in Japan. Interestingly, average HDL levels in Japan have increased further in the past two decades, and are markedly higher than in Western populations. The reasons and consequences for public health of this increase are still unknown. Simulation for the efficacy of raising HDL cholesterol predicts a decrease in CHD of 70% in Japan, greater than the extent by reducing low-density lipoprotein cholesterol predicted by simulation or achieved in a statin trial. On the other hand, a substantial portion of hyperalphalipoproteinemic population in Japan is accounted for by genetic deficiency of cholesteryl ester transfer protein (CETP), which is also commonly unique in East Asian populations. It is still controversial whether CETP mutations are antiatherogenic. Hepatic Schistosomiasis is proposed as a potential screening factor for historic accumulation of CETP deficiency in East Asia. PMID:25849946

  10. Unique Features of High-Density Lipoproteins in the Japanese: In Population and in Genetic Factors

    PubMed Central

    Yokoyama, Shinji

    2015-01-01

    Despite its gradual increase in the past several decades, the prevalence of atherosclerotic vascular disease is low in Japan. This is largely attributed to difference in lifestyle, especially food and dietary habits, and it may be reflected in certain clinical parameters. Plasma high-density lipoprotein (HDL) levels, a strong counter risk for atherosclerosis, are indeed high among the Japanese. Accordingly, lower HDL seems to contribute more to the development of coronary heart disease (CHD) than an increase in non-HDL lipoproteins at a population level in Japan. Interestingly, average HDL levels in Japan have increased further in the past two decades, and are markedly higher than in Western populations. The reasons and consequences for public health of this increase are still unknown. Simulation for the efficacy of raising HDL cholesterol predicts a decrease in CHD of 70% in Japan, greater than the extent by reducing low-density lipoprotein cholesterol predicted by simulation or achieved in a statin trial. On the other hand, a substantial portion of hyperalphalipoproteinemic population in Japan is accounted for by genetic deficiency of cholesteryl ester transfer protein (CETP), which is also commonly unique in East Asian populations. It is still controversial whether CETP mutations are antiatherogenic. Hepatic Schistosomiasis is proposed as a potential screening factor for historic accumulation of CETP deficiency in East Asia. PMID:25849946

  11. High-speed AFM for 1x node metrology and inspection: Does it damage the features?

    NASA Astrophysics Data System (ADS)

    Sadeghian, Hamed; van den Dool, Teun C.; Uziel, Yoram; Bar Or, Ron

    2015-03-01

    This paper aims at unraveling the mystery of damage in high speed AFMs for 1X node and below. With the device dimensions moving towards the 1X node and below, the semiconductor industry is rapidly approaching the point where existing metrology, inspection and review tools face huge challenges in terms of resolution, the ability to resolve 3D, and throughput. In this paper, we critically asses the important issue of damage in high speed AFM for metrology and inspection of semiconductor wafers. The issues of damage in four major scanning modes (contact mode, tapping mode, non-contact mode, and peak force tapping mode) are described to show which modes are suitable for which applications and which conditions are damaging. The effects of all important scanning parameters on resulting damage are taken into account for materials such as silicon, photoresists and low K materials. Finally, we recommend appropriate scanning parameters and conditions for several use cases (FinFET, patterned photoresist, HAR structures) that avoid exceeding a critical contact stress such that sample damage is minimized. In conclusion, we show using our theoretical analysis that selecting parameters that exceed the target contact stress, indeed leads to significant damage. This method provides AFM users for metrology with a better understanding of contact stresses and enables selection of AFM cantilevers and experimental parameters that prevent sample damage.

  12. Features of brittle damages and hydrogen impregnation of high-pressure boiler tube metal

    SciTech Connect

    Vainman, A.B.; Smiyan, O.D.; Girnyi, S.I.; Kostyuchenko, N.P.; Vasilik, A.V.; Melekhov, R.K.

    1988-01-01

    A significant number of failures encountered in high-pressure steam boilers of thermal electric power stations are caused by damage of the tubes of the steam superheaters, which are made of 20, 12Kh1MF, 12Kh2MFSR and 12Kh18N10T steels. A statistical analysis made by the authors determined that a large number of the failures result from the action of hydrogen on the tubes. In this paper, the mechanisms of hydrogen diffusion, corrosion crack propagation, and brittle failure for steam superheater tubes were analyzed. Hydrogen-related intergranular cracking and thermal fatigue were assessed for the tube steels. It was concluded that hydrogen had a significant effect on processes of crack origin and propagation both in the superheater and in the unheated tubes of the boilers.

  13. Discovery of a Highly Magnetic White Dwarf with Strong Carbon Features

    NASA Astrophysics Data System (ADS)

    Schmidt, Gary D.; Liebert, James; Harris, Hugh C.; Dahn, Conard C.; Leggett, S. K.

    1999-02-01

    Systematic follow-up of high proper-motion stars has identified a new cool magnetic white dwarf that displays a series of spectacular absorption bands in the range 4200-6500 Å. The spectrum bears a striking resemblance to that of LP 790-29, a magnetic DQ star dominated by what are apparently Zeeman-shifted Swan bands of C2. However, key differences in the detailed spectra, polarization, and temperature of the two stars indicate that instead LHS 2229 may represent the first case of a magnetic ``peculiar'' DQ white dwarf, where absorption in the optical is produced by C2H or another carbon-hydrogen compound. Crude arguments suggest that the field strength on LHS 2229 is in the neighborhood of 108 G. BVI photometry proves to be effective in identifying such peculiar stars, since they lie well outside the main white dwarf sequence in a color-color diagram.

  14. Microstructure features of high-entropy equiatomic cast AlCrFeCoNiCu alloys

    NASA Astrophysics Data System (ADS)

    Ivchenko, M. V.; Pushin, V. G.; Uksusnikov, A. N.; Wanderka, N.

    2013-06-01

    The structural and phase transformations that take place in the cast high-entropy equiatomic alloy AlCrFeCoNiCu after solidification, homogenizing heat treatment, and cooling have been studied. Analytical transmission microscopy, scanning electron microscopy, X-ray energy dispersive spectroscopy, and X-ray diffraction analysis were used to conduct the studies. The elastic modulus, nano-, and microhardness have been measured. The alloy decomposition has been found to occur with the precipitation of no less than six nanoscale phases with different morphologies, structures ( A2, B2, L12), and chemical compositions. All the nanophases are multicomponent solid solutions enriched with several elements, which indicates the pronounced elemental and phase nanomodulation over the alloy volume.

  15. A cosmic ray super high multicore family event. 1: Experiment and general features

    NASA Technical Reports Server (NTRS)

    Ren, J. R.; Kuang, H. H.; Huo, A. X.; Lu, S. L.; Su, S.; Wang, Y. X.; Xue, Y. G.; Wang, C. R.; He, M.; Zhang, N. J.

    1985-01-01

    Information on the fragmentation region in super high energy hadronic interactions can be obtained through the observations of gamma-ray families produced by cosmic rays. Gamma-ray families with the sum of E sub gamma or 1000 TeV are receiving increasing interests in emulsion chamber experiments. There exist some complications caused by the superposition of nuclear and electromagnetic cascades and the uncertainty in the nature of the primary particles. These complications usually make the conclusions drawn from various interesting phenomena observed in family events not so definite. An interesting family event KO E19, which is likely to have suffered only very slight disturbances is described. It was found in the Mt. Kambala emulsion chamber experiment. The production height of the event is determined to be H=(70 + or - 30)m and some conclusions are given.

  16. DE/ISIS conjunction comparisons of high-latitude electron density features

    NASA Technical Reports Server (NTRS)

    Hoegy, Walter R.; Benson, Robert F.

    1988-01-01

    This paper presents a comparison between the ISIS-1 and -2 topside sounder measurements of electron number density, N(e), with the in situ ion and N(e) measurements by the Langmuir probe aboard the Dynamics Explorer 2 (DE 2) during four high-latitude ISIS/DE magnetic field-aligned conjunctions. The ISIS-derived N(e) values, even at the greatest distance from the sounder, were found to agree with the Langmuir probe measurements to within about 30 percent over a density range of more than two decades on three of the four comparisons; the fourth comparison which included data with strong N(e) irregularities, showed a difference of 60 percent.

  17. Transmission electron microscopy specimen preparation perpendicular to the long axis of high aspect ratio features

    SciTech Connect

    Irwin, R. B.; Anciso, A.; Jones, P. J.; Glenn, A. L.; Williams, B. L.; Sridhar, S.; Arshad, S.

    2009-11-15

    A new variation of transmission electron microscopy (TEM) specimen preparation is introduced. By thinning a tall high aspect ratio structure perpendicular to the long dimension (i.e., from the side) rather than from perpendicular to the short dimension (either the top or the bottom), it is possible to obtain a more uniformly thin TEM specimen over the entire long dimension of the structure. This article will describe the rational for this variation in specimen preparation. The necessary modifications of four different specimen preparation methods (in situ lift-out, traditional H-bar, ex situ lift-out, and tripod polishing) will be discussed and images of specimens obtained by both of these first two methods will be shown. Additional potential advantages and other applications of this specimen preparation method will be covered.

  18. KCd spectral features in K+Na+Cd+Ar high pressure lamp.

    NASA Astrophysics Data System (ADS)

    Džimbeg-Malčić, V.; Beuc, R.; Pichler, G.

    We present the results of spectroscopic studies of the high pressure K-Na-Cd-Ar discharge lamp. They were compared with the results of K-Hg-Ar discharge lamp. The interesting part of the spectrum was the region around the second resonance potassium line (5p-4s transition) at 404.4 nm and 404.7 nm. There we observed a group of satellite bands which we attribute to the KCd excimer transitions. In the red part of the spectrum a broad KCd diffuse band was observed with peaks at 630 nm and 640 nm. From the comparison with the KHg system, we ascribe these bands to the 22P-X2 S+ electronic transitions.

  19. Structural features and mechanical properties of austenitic Hadfield steel after high-pressure torsion and subsequent high-temperature annealing

    NASA Astrophysics Data System (ADS)

    Tukeeva, M. S.; Melnikov, E. V.; Maier, H. J.; Astafurova, E. G.

    2012-06-01

    Mechanisms of structure fragmentation and strengthening of single crystals of a Hadfield steel after warm torsion under high-pressure torsion (HPT) and subsequent annealing in a temperature range of 400-800°C have been studied. Multiple twinning and formation of ultrafine carbides upon HPT at 400°C ( P = 5 GPa) promote rapid fragmentation of the microstructure. They are responsible for the high mechanical properties of the steel after HPT and the thermal stability of the microstructure up to an annealing temperature of 500°C.

  20. Pathobiological features of a novel, highly pathogenic avian influenza A(H5N8) virus.

    PubMed

    Kim, Young-Il; Pascua, Philippe Noriel Q; Kwon, Hyeok-Il; Lim, Gyo-Jin; Kim, Eun-Ha; Yoon, Sun-Woo; Park, Su-Jin; Kim, Se Mi; Choi, Eun-Ji; Si, Young-Jae; Lee, Ok-Jun; Shim, Woo-Sub; Kim, Si-Wook; Mo, In-Pil; Bae, Yeonji; Lim, Yong Taik; Sung, Moon Hee; Kim, Chul-Joong; Webby, Richard J; Webster, Robert G; Choi, Young Ki

    2014-10-01

    The endemicity of highly pathogenic avian influenza (HPAI) A(H5N1) viruses in Asia has led to the generation of reassortant H5 strains with novel gene constellations. A newly emerged HPAI A(H5N8) virus caused poultry outbreaks in the Republic of Korea in 2014. Because newly emerging high-pathogenicity H5 viruses continue to pose public health risks, it is imperative that their pathobiological properties be examined. Here, we characterized A/mallard duck/Korea/W452/2014 (MDk/W452(H5N8)), a representative virus, and evaluated its pathogenic and pandemic potential in various animal models. We found that MDk/W452(H5N8), which originated from the reassortment of wild bird viruses harbored by migratory waterfowl in eastern China, replicated systemically and was lethal in chickens, but appeared to be attenuated, albeit efficiently transmitted, in ducks. Despite predominant attachment to avian-like virus receptors, MDk/W452(H5N8) also exhibited detectable human virus-like receptor binding and replicated in human respiratory tract tissues. In mice, MDk/W452(H5N8) was moderately pathogenic and had limited tissue tropism relative to previous HPAI A(H5N1) viruses. It also induced moderate nasal wash titers in inoculated ferrets; additionally, it was recovered in extrapulmonary tissues and one of three direct-contact ferrets seroconverted without shedding. Moreover, domesticated cats appeared to be more susceptible than dogs to virus infection. With their potential to become established in ducks, continued circulation of A(H5N8) viruses could alter the genetic evolution of pre-existing avian poultry strains. Overall, detailed virological investigation remains a necessity given the capacity of H5 viruses to evolve to cause human illness with few changes in the viral genome. PMID:26038499

  1. The Structure, Water Budget, and Radiational Features of a High-Latitude Warm Front.

    NASA Astrophysics Data System (ADS)

    Hanesiak, John M.; Stewart, Ronald E.; Szeto, Kit K.; Hudak, David R.; Leighton, Henry G.

    1997-06-01

    On 30 September 1994 an Arctic low pressure system passed over the southern Beaufort Sea area of northern Canada and research aircraft observations were made within and around the warm front of the storm. This study is unique in that the warm front contained subzero centigrade temperatures across the entire frontal region. The overall structure of the warm front and surrounding region was similar to midlatitude storms; however, the precipitation rates, liquid water content magnitudes, horizontal and vertical winds, vertical wind shear, turbulence, and thermal advection were very weak. In addition, a low-level jet and cloud bands were aligned parallel to the warm front, near-neutral stability occurred within and around the front, and conditional symmetric instability was likely occurring. A steep frontal region resulted from strong Coriolis influences that in turn limited the amount of cloud and precipitation ahead of the system. The precipitation efficiency of the storm was high (60%) but is believed to be highly dependent on the stage of development. The mesoscale frontogenetic forcing was primarily controlled by the tilting of isentropic surfaces with confluence/convergence being the secondary influence. Sublimation contributions may have been large in the earlier stages of storm development. Satellite and aircraft radiometers underestimated cloud top heights by as much as 4 km and this was mostly due to the near transparency of the lofted ice layer in the upper portion of the storm. Maximum surface solar radiation deficits ranged between 91 W m2 and 187 W m2 at two surface observing sites. This common type of cloud system must have a major impact on the water and energy cycles of northern Canada in the autumn and therefore must be well accounted for within climate models.

  2. Collecting Inexpensive High Resolution Aerial and Stereo Images of Small- to Mid-Scale Geomorphic and Tectonic Features

    NASA Astrophysics Data System (ADS)

    Wheelwright, R. J.; White, W. S.; Willis, J. B.

    2010-12-01

    Methods for collecting accurate, mm- to cm-scale stereoscopic aerial imagery of both small- and mid-scale geomorphic features are developed for a one-time cost of under $1500. High resolution aerial images are valuable for documenting and analyzing small- to mid-scale geomorphic and tectonic features. However, collecting images of mid-scale features such as landslides, rock glaciers, fault scarps, and cinder cones is expensive and makes studies that rely on high resolution repeat imagery prohibitive for undergraduate geology departments with limited budgets. In addition to cost, collecting images of smaller scale geomorphic features such as gravel bars is often impeded by overhanging vegetation or other features in the immediate environment that make impractical the collection of aerial images using standard airborne techniques. The methods provide high resolution stereo photos suitable for image processing and stereographic analysis; the images are potentially suitable for change analyses, velocity tracking, and construction of lidar-resolution digital elevation models. We developed two techniques. The technique suitable for small-scale features (such as gravel bars) utilizes two Nikon D3000 digital single-lens reflex (DSLR) cameras attached to a system of poles that suspends the cameras at a height of 4 meters with a variable camera separation of 0.6 to 0.9 m. The poles are oriented such that they do not appear in the photographs. The cameras are simultaneously remotely activated to collect stereo pairs at a resolution of 64 pixels/cm2 (pixel length is 1.2 mm). Ground control on the images is provided by pegs placed 5 meters apart, GPS positioning, and a meter-stick included in each photograph. Initial photo data gathered of a gravel bar on the Henrys Fork of the Snake River, north of Rexburg, Idaho is sharp and readily segmented using the MatLab-based CLASTS image processing algorithm. The technique developed for imaging mid-scale features (such as cinder cones) consists of a tethered weather balloon with a gimbal that keeps the camera oriented vertically. A single DSLR camera is suspended at an elevation of approximately 45 m. The camera is operated via a radio-controlled apparatus that enables the user to view the area being photographed prior to activating the shutter. The balloon is physically moved to capture the second image of the stereo pair. Resolution of the images is 3600 pixels/m2 (pixel length is 1.6 cm). The techniques demonstrate that collecting high-resolution stereographic aerial photographs can be accomplished on a limited budget. The techniques and equipment will be used to collect repeat images for several multi-year studies, including monitoring gravel bar evolution, vector tracking of landslide and rock glacier motion, and monitoring fault scarp and cinder cone degradation.

  3. A spectral-structural bag-of-features scene classifier for very high spatial resolution remote sensing imagery

    NASA Astrophysics Data System (ADS)

    Zhao, Bei; Zhong, Yanfei; Zhang, Liangpei

    2016-06-01

    Land-use classification of very high spatial resolution remote sensing (VHSR) imagery is one of the most challenging tasks in the field of remote sensing image processing. However, the land-use classification is hard to be addressed by the land-cover classification techniques, due to the complexity of the land-use scenes. Scene classification is considered to be one of the expected ways to address the land-use classification issue. The commonly used scene classification methods of VHSR imagery are all derived from the computer vision community that mainly deal with terrestrial image recognition. Differing from terrestrial images, VHSR images are taken by looking down with airborne and spaceborne sensors, which leads to the distinct light conditions and spatial configuration of land cover in VHSR imagery. Considering the distinct characteristics, two questions should be answered: (1) Which type or combination of information is suitable for the VHSR imagery scene classification? (2) Which scene classification algorithm is best for VHSR imagery? In this paper, an efficient spectral-structural bag-of-features scene classifier (SSBFC) is proposed to combine the spectral and structural information of VHSR imagery. SSBFC utilizes the first- and second-order statistics (the mean and standard deviation values, MeanStd) as the statistical spectral descriptor for the spectral information of the VHSR imagery, and uses dense scale-invariant feature transform (SIFT) as the structural feature descriptor. From the experimental results, the spectral information works better than the structural information, while the combination of the spectral and structural information is better than any single type of information. Taking the characteristic of the spatial configuration into consideration, SSBFC uses the whole image scene as the scope of the pooling operator, instead of the scope generated by a spatial pyramid (SP) commonly used in terrestrial image classification. The experimental results show that the whole image as the scope of the pooling operator performs better than the scope generated by SP. In addition, SSBFC codes and pools the spectral and structural features separately to avoid mutual interruption between the spectral and structural features. The coding vectors of spectral and structural features are then concatenated into a final coding vector. Finally, SSBFC classifies the final coding vector by support vector machine (SVM) with a histogram intersection kernel (HIK). Compared with the latest scene classification methods, the experimental results with three VHSR datasets demonstrate that the proposed SSBFC performs better than the other classification methods for VHSR image scenes.

  4. Novel digital diffractive tags integrating anti-counterfeiting, tamper-evident, and high-density WORM data storage features

    NASA Astrophysics Data System (ADS)

    Boisdur, Enrick; Kress, Bernard

    2010-05-01

    Embossed holographic tags for security and anti-counterfeiting applications are being used by industry since many years. However, such elements are not very effective since the detector is usually the human eye, and provides therefore around 80% effective counterfeiting protection of the tag. We present a novel holographic anticounterfeiting technology which provides 99.999% protection against tag counterfeiting. Horus Technologies develops such holographic tags, which include several layers of increasingly secure optical features, from standard visual holographic patterns and OVIDs (Optical Variable Imaging Devices), to micro-holographic text, down to covert features such as encrypted high resolution holographic 1d, 2d and 3d bar codes. We also demonstrate the potential of providing anti-tamper functionality on the same tag, for packaging security (especially for medical packaging). Finally, we demonstrate that more than 1Mb/square mm of digital data can be stored and encrypted on these same tags. A specific low cost laser based reader is developed to read the various security feature of such hybrid universal holographic tags. We also present a way to change and update the encrypted data in the tag in a similar way to RFID tags. Finally, we show a cost effective technique to replicate these structures in volume by roll-to-toll embossing, and even direct by glass molding within the package itself (bottle, vial, etc,..).

  5. The endocrine dyscrasia that accompanies menopause and andropause induces aberrant cell cycle signaling that triggers re-entry of post-mitotic neurons into the cell cycle, neurodysfunction, neurodegeneration and cognitive disease.

    PubMed

    Atwood, Craig S; Bowen, Richard L

    2015-11-01

    This article is part of a Special Issue "SBN 2014". Sex hormones are physiological factors that promote neurogenesis during embryonic and fetal development. During childhood and adulthood these hormones support the maintenance of brain structure and function via neurogenesis and the formation of dendritic spines, axons and synapses required for the capture, processing and retrieval of information (memories). Not surprisingly, changes in these reproductive hormones that occur with menopause and during andropause are strongly correlated with neurodegeneration and cognitive decline. In this connection, much evidence now indicates that Alzheimer's disease (AD) involves aberrant re-entry of post-mitotic neurons into the cell cycle. Cell cycle abnormalities appear very early in the disease, prior to the appearance of plaques and tangles, and explain the biochemical, neuropathological and cognitive changes observed with disease progression. Intriguingly, a recent animal study has demonstrated that induction of adult neurogenesis results in the loss of previously encoded memories while decreasing neurogenesis after memory formation during infancy mitigated forgetting. Here we review the biochemical, epidemiological and clinical evidence that alterations in sex hormone signaling associated with menopause and andropause drive the aberrant re-entry of post-mitotic neurons into an abortive cell cycle that leads to neurite retraction, neuron dysfunction and neuron death. When the reproductive axis is in balance, gonadotropins such as luteinizing hormone (LH), and its fetal homolog, human chorionic gonadotropin (hCG), promote pluripotent human and totipotent murine embryonic stem cell and neuron proliferation. However, strong evidence supports menopausal/andropausal elevations in the LH:sex steroid ratio as driving aberrant mitotic events. These include the upregulation of tumor necrosis factor; amyloid-β precursor protein processing towards the production of mitogenic Aβ; and the activation of Cdk5, a key regulator of cell cycle progression and tau phosphorylation (a cardinal feature of both neurogenesis and neurodegeneration). Cognitive and biochemical studies confirm the negative consequences of a high LH:sex steroid ratio on dendritic spine density and human cognitive performance. Prospective epidemiological and clinical evidence in humans supports the premise that rebalancing the ratio of circulating gonadotropins:sex steroids reduces the incidence of AD. Together, these data support endocrine dyscrasia and the subsequent loss of cell cycle control as an important etiological event in the development of neurodegenerative diseases including AD, stroke and Parkinson's disease. PMID:26188949

  6. High-Resolution Transcriptome Maps Reveal Strain-Specific Regulatory Features of Multiple Campylobacter jejuni Isolates

    PubMed Central

    Förstner, Konrad U.; Heidrich, Nadja; Reinhardt, Richard; Nieselt, Kay; Sharma, Cynthia M.

    2013-01-01

    Campylobacter jejuni is currently the leading cause of bacterial gastroenteritis in humans. Comparison of multiple Campylobacter strains revealed a high genetic and phenotypic diversity. However, little is known about differences in transcriptome organization, gene expression, and small RNA (sRNA) repertoires. Here we present the first comparative primary transcriptome analysis based on the differential RNA–seq (dRNA–seq) of four C. jejuni isolates. Our approach includes a novel, generic method for the automated annotation of transcriptional start sites (TSS), which allowed us to provide genome-wide promoter maps in the analyzed strains. These global TSS maps are refined through the integration of a SuperGenome approach that allows for a comparative TSS annotation by mapping RNA–seq data of multiple strains into a common coordinate system derived from a whole-genome alignment. Considering the steadily increasing amount of RNA–seq studies, our automated TSS annotation will not only facilitate transcriptome annotation for a wider range of pro- and eukaryotes but can also be adapted for the analysis among different growth or stress conditions. Our comparative dRNA–seq analysis revealed conservation of most TSS, but also single-nucleotide-polymorphisms (SNP) in promoter regions, which lead to strain-specific transcriptional output. Furthermore, we identified strain-specific sRNA repertoires that could contribute to differential gene regulation among strains. In addition, we identified a novel minimal CRISPR-system in Campylobacter of the type-II CRISPR subtype, which relies on the host factor RNase III and a trans-encoded sRNA for maturation of crRNAs. This minimal system of Campylobacter, which seems active in only some strains, employs a unique maturation pathway, since the crRNAs are transcribed from individual promoters in the upstream repeats and thereby minimize the requirements for the maturation machinery. Overall, our study provides new insights into strain-specific transcriptome organization and sRNAs, and reveals genes that could modulate phenotypic variation among strains despite high conservation at the DNA level. PMID:23696746

  7. Features of High-Temperature Calibration of W/Re Thermocouples

    NASA Astrophysics Data System (ADS)

    Ulanovskiy, A.; Edler, F.; Fischer, J.; Oleynikov, P.; Zaytsev, P.; Pokhodun, A.

    2015-03-01

    Investigations of Type A (W-5 %Re/W-20 %Re) thermocouples were performed at several laboratories to validate their reference function before its standardization in the new issue of the international standard IEC 60584. The Type A thermocouples investigated were equipped with sealed protection tubes made of sapphire which were filled with an inert gas (argon). The investigations at Russian laboratories were performed mainly in carbon-free high-temperature furnaces. The calibration results obtained in the temperature range (600 to 1850) in the carbon-free environment were within % tolerance limits and confirmed the suitability of Type A thermocouples for industrial applications. In contrast, the Type A thermocouple 89/95-P investigated at PTB (Germany) was exposed to a carbon environment when annealed at and when it was calibrated at metal-carbon eutectic (Me-C) fixed points. Measurements made at Me-C fixed points did not deviate from the reference function by more than about 6 K at the first stage when temperatures were below . However, the inhomogeneity of the thermoelements increased continuously after the calibration at the Me-C eutectic fixed points. The additional measurements at the peritectic fixed point () resulted in a continuous emf drift to deviations from the reference function of about (100 to 150) which corresponds to a temperature equivalent of about (9 to 14) K. The thermoelectric stability and homogeneity of the thermocouple 89/95-P during these investigations was checked by repeated measurements at the freezing point of copper ().

  8. Features of primary damage by high energy displacement cascades in concentrated Ni-based alloys

    NASA Astrophysics Data System (ADS)

    Béland, Laurent Karim; Lu, Chenyang; Osetskiy, Yuri N.; Samolyuk, German D.; Caro, Alfredo; Wang, Lumin; Stoller, Roger E.

    2016-02-01

    Alloying of Ni with Fe or Co has been shown to reduce primary damage production under ion irradiation. Similar results have been obtained from classical molecular dynamics simulations of 1, 10, 20, and 40 keV collision cascades in Ni, NiFe, and NiCo. In all cases, a mix of imperfect stacking fault tetrahedra, faulted loops with a 1/3⟨111⟩ Burgers vector, and glissile interstitial loops with a 1/2⟨110⟩ Burgers vector were formed, along with small sessile point defect complexes and clusters. Primary damage reduction occurs by three mechanisms. First, Ni-Co, Ni-Fe, Co-Co, and Fe-Fe short-distance repulsive interactions are stiffer than Ni-Ni interactions, which lead to a decrease in damage formation during the transition from the supersonic ballistic regime to the sonic regime. This largely controls final defect production. Second, alloying decreases thermal conductivity, leading to a longer thermal spike lifetime. The associated annealing reduces final damage production. These two mechanisms are especially important at cascades energies less than 40 keV. Third, at the higher energies, the production of large defect clusters by subcascades is inhibited in the alloys. A number of challenges and limitations pertaining to predictive atomistic modeling of alloys under high-energy particle irradiation are discussed.

  9. Features of primary damage by high energy displacement cascades in concentrated Ni-based alloys

    DOE PAGESBeta

    Béland, Laurent Karim; Lu, Chenyang; Osetskiy, Yuri N.; Samolyuk, German D.; Caro, Alfredo; Wang, Lumin; Stoller, Roger E.

    2016-02-25

    Alloying of Ni with Fe or Co reduces primary damage production under ion irradiation. Similar results have been obtained from classical molecular dynamics simulations of 1, 10, 20, and 40 keV collision cascades in Ni, NiFe, and NiCo. In all cases, a mix of imperfect stacking fault tetrahedra, faulted loops with a 1/3 {111} Burgers vector, and glissile interstitial loops with a 1/2 {110} Burgers vector were formed, along with small sessile point defect complexes and clusters. Primary damage reduction occurs by three mechanisms. First, Ni-Co, Ni-Fe, Co-Co, and Fe-Fe short-distance repulsive interactions are stiffer than Ni-Ni interactions, which leadmore » to a decrease in damage formation during the transition from the supersonic ballistic regime to the sonic regime. This largely controls final defect production. Second, alloying decreases thermal conductivity, leading to a longer thermal spike lifetime. The associated annealing reduces final damage production. These two mechanisms are especially important at cascades energies less than 40 keV. Third, at the higher energies, the production of large defect clusters by subcascades is inhibited in the alloys. A number of challenges and limitations pertaining to predictive atomistic modeling of alloys under high-energy particle irradiation are discussed.« less

  10. High-resolution continuously acquired peripheral MR angiography featuring partial parallel imaging GRAPPA.

    PubMed

    Zenge, Michael O; Vogt, Florian M; Brauck, Katja; Jökel, Michaela; Barkhausen, Joerg; Kannengiesser, Stephan; Ladd, Mark E; Quick, Harald H

    2006-10-01

    Continuously-moving-table MRI, in contrast to traditional multistation techniques, potentially can improve the scan time efficiency of whole-body applications and provide seamless images of an extended field of view (FOV). Contrast-enhanced MR angiography (CE-MRA) in particular requires high spatial resolution and at the same time has rigid scan time constraints due to the limited arterial contrast window. In this study a reconstruction method for continuously acquired 3D data sets during table movement was combined with a self-calibrated partial parallel imaging algorithm (generalized autocalibrating partially parallel acquisitions (GRAPPA)). The method was applied to peripheral CE-MRA and compared with a standard continuously-moving-table MRA protocol. The gain in scan time was used to increase the data acquisition matrix and decrease the slice thickness. The method was evaluated in five healthy volunteers and applied to one patient with peripheral arterial occlusive disease (PAOD). The protocols were intraindividually compared with respect to the signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) in selected vessel segments, as well as overall vessel depiction. The combination of the continuously-moving-table technique with parallel imaging enabled the acquisition of seamless peripheral 3D MRA with increased resolution and an overall crisper appearance. PMID:16964615

  11. High amplitude theta wave bursts: a novel electroencephalographic feature of rem sleep and cataplexy.

    PubMed

    Lo Martire, Viviana Carmen; Bastianini, Stefano; Berteotti, Chiara; Silvani, Alessandro; Zoccoli, Giovanna

    2015-09-01

    High amplitude theta wave bursts (HATs) were originally described during REMS and cataplexy in ORX-deficient mice as a novel neurophysiological correlate of narcolepsy (Bastianini et al., 2012). This finding was replicated the following year by Vassalli et al. in both ORX-deficient narcoleptic mice and narcoleptic children during cataplexy episodes (Vassalli et al., 2013). The relationship between HATs and narcolepsy-cataplexy in mice and patients indicates that the lack of ORX peptides is responsible for this abnormal EEG activity, the physiological meaning of which is still unknown. This review aimed to explore different phasic EEG events previously described in the published literature in order to find analogies and differences with HATs observed in narcoleptic mice and patients. We found similarities in terms of morphology, frequency and duration between HATs and several physiological (mu and wicket rhythms, sleep spindles, saw-tooth waves) or pathological (SWDs, HVSs, bursts of polyphasic complexes EEG complexes reported in a mouse model of CJD, and BSEs) EEG events. However, each of these events also shows significant differences from HATs, and thus cannot be equaled to them. The available evidence thus suggests that HATs are a novel neurophysiological phenomenon. Further investigations on HATs are required in order to investigate their physiological meaning, to individuate their brain structure(s) of origin, and to clarify the neural circuits involved in their manifestation. PMID:26742662

  12. Features of oxide layer formation in high-aspect slot structures by means of MOCVD

    NASA Astrophysics Data System (ADS)

    Shevtsov, Yuri V.; Kuchumov, Boris М.; Kruchinin, Vladimir N.; Spesivtsev, Evgeni V.; Golovnev, Igor F.; Igumenov, Igor К.

    2015-03-01

    Processes were studied concerning the deposition of Hf and Mg oxide layers in the slot structures with the aspect ratio values from 30 to 500 by means of a pulsed MOCVD technique with a discrete components dosing. For assembling the slot structures, different combinations of materials have been used such as Si/Si, Si/glass, glass/chromium patterned glass, and Si/chromium patterned glass. The layers were characterized by means of XRD, XPS, and SEM methods. The thickness profiles of deposited films were measured using a high spatial resolution laser ellipsometry. It has been demonstrated that the radial distribution profiles of the thickness are determined by the process temperature, the aspect ratio value and the type of precursor. In contrast to HfO2 the layers of MgO with decreasing process temperature demonstrate change in the shape of layer profile distribution from a bowl-like shape to a dome-like one, with an increase of the film thickness at the center of the substrate. An image transfer process in the slot structure wherein one of the glass substrates is chromium-patterned has been studied within a wide range of experimental parameters. In order to describe these effects there has been a hybrid model proposed involving a combination of molecular dynamics and Monte Carlo methods. All the results obtained are discussed in detail.

  13. High-precision, automated integration of multiple isothermal titration calorimetric thermograms: new features of NITPIC.

    PubMed

    Scheuermann, Thomas H; Brautigam, Chad A

    2015-04-01

    Isothermal titration calorimetry (ITC) has become a standard and widely available tool to measure the thermodynamic parameters of macromolecular associations. Modern applications of the method, including global analysis and drug screening, require the acquisition of multiple sets of data; sometimes these data sets number in the hundreds. Therefore, there is a need for quick, precise, and automated means to process the data, particularly at the first step of data analysis, which is commonly the integration of the raw data to yield an interpretable isotherm. Herein, we describe enhancements to an algorithm that previously has been shown to provide an automated, unbiased, and high-precision means to integrate ITC data. These improvements allow for the speedy and precise serial integration of an unlimited number of ITC data sets, and they have been implemented in the freeware program NITPIC, version 1.1.0. We present a comprehensive comparison of the performance of this software against an older version of NITPIC and a current version of Origin, which is commonly used for integration. The new methods recapitulate the excellent performance of the previous versions of NITPIC while speeding it up substantially, and their precision is significantly better than that of Origin. This new version of NITPIC is therefore well suited to the serial integration of many ITC data sets. PMID:25524420

  14. High-precision, automated integration of multiple isothermal titration calorimetric thermograms: new features of NITPIC

    PubMed Central

    Scheuermann, Thomas H.; Brautigam, Chad A.

    2014-01-01

    Isothermal titration calorimetry (ITC) has become a standard and widely available tool to measure the thermodynamic parameters of macromolecular associations. Modern applications of the method, including global analysis and drug screening, require the acquisition of multiple sets of data; sometimes these data sets number in the hundreds. Therefore, there is a need for quick, precise, and automated means to process the data, particularly at the first step of data analysis, which is commonly the integration of the raw data to yield an interpretable isotherm. Herein, we describe enhancements to an algorithm that previously has been shown to provide an automated, unbiased, and high-precision means to integrate ITC data. These improvements allow for the speedy and precise serial integration of an unlimited number of ITC data sets, and they have been implemented in the freeware program NITPIC, version 1.1.0. We present a comprehensive comparison of the performance of this software against an older version of NITPIC and a current version of Origin, which is commonly used for integration. The new methods recapitulate the excellent performance of the previous versions of NITPIC while speeding it up substantially, and their precision is significantly better than that of Origin. This new version of NITPIC is therefore well suited to the serial integration of many ITC data sets. PMID:25524420

  15. Microcephaly Disease Gene Wdr62 Regulates Mitotic Progression of Embryonic Neural Stem Cells and Brain Size

    PubMed Central

    Chen, Jian-Fu; Zhang, Ying; Wilde, Jonathan; Hansen, Kirk; Lai, Fan; Niswander, Lee

    2014-01-01

    Human genetic studies have established a link between a class of centrosome proteins and microcephaly. Current studies of microcephaly focus on defective centrosome/spindle orientation. Mutations in WDR62 are associated with microcephaly and other cortical abnormalities in humans. Here we create a mouse model of Wdr62 deficiency and find that the mice exhibit reduced brain size due to decreased neural progenitor cells (NPCs). Wdr62 depleted cells show spindle instability, spindle assembly checkpoint (SAC) activation, mitotic arrest and cell death. Mechanistically, Wdr62 associates and genetically interacts with Aurora A to regulate spindle formation, mitotic progression and brain size. Our results suggest that Wdr62 interacts with Aurora A to control mitotic progression, and loss of these interactions leads to mitotic delay and cell death of NPCs, which could be a potential cause of human microcephaly. PMID:24875059

  16. Mitotic rounding alters cell geometry to ensure efficient bipolar spindle formation.

    PubMed

    Lancaster, Oscar M; Le Berre, Maël; Dimitracopoulos, Andrea; Bonazzi, Daria; Zlotek-Zlotkiewicz, Ewa; Picone, Remigio; Duke, Thomas; Piel, Matthieu; Baum, Buzz

    2013-05-13

    Accurate animal cell division requires precise coordination of changes in the structure of the microtubule-based spindle and the actin-based cell cortex. Here, we use a series of perturbation experiments to dissect the relative roles of actin, cortical mechanics, and cell shape in spindle formation. We find that, whereas the actin cortex is largely dispensable for rounding and timely mitotic progression in isolated cells, it is needed to drive rounding to enable unperturbed spindle morphogenesis under conditions of confinement. Using different methods to limit mitotic cell height, we show that a failure to round up causes defects in spindle assembly, pole splitting, and a delay in mitotic progression. These defects can be rescued by increasing microtubule lengths and therefore appear to be a direct consequence of the limited reach of mitotic centrosome-nucleated microtubules. These findings help to explain why most animal cells round up as they enter mitosis. PMID:23623611

  17. Physical lengths of meiotic and mitotic gene conversion tracts in Saccharomyces cerevisiae

    SciTech Connect

    Judd, S.R.; Petes, T.D.

    1988-03-01

    Physical lengths of gene conversion tracts for meiotic and mitotic conversions were examined, using the same diploid yeast strain in all experiments. This strain is heterozygous for a mutation in the URA3 gene as well as closely linked restriction site markers. In cells that had a gene conversion event at the URA3 locus, it was determined by Southern analysis which of the flanking heterozygous restriction sites had co-converted. It was found that mitotic conversion tracts were longer on the average than meiotic tracts. About half of the tracts generated by spontaneous mitotic gene conversion included heterozygous markers 4.2 kb apart; none of the meiotic conversions included these markers. Stimulation of mitotic gene conversion by ultraviolet light or methylmethanesulfonate had no obvious effect on the size or distribution of the tracts. Almost all conversion tracts were continuous.

  18. Association of Chromosome Loss with Centromere-Adjacent Mitotic Recombination in a Yeast Disomic Haploid

    PubMed Central

    Campbell, D. A.; Fogel, S.

    1977-01-01

    Experiments designed to characterize the association between disomic chromosome loss and centromere-adjacent mitotic recombination were performed. Mitotic gene convertants were selected at two heteroallelic sites on the left arm of disomic chromosome III and tested for coincident chromosome loss. The principal results are: (1) Disomic chromosome loss is markedly enhanced (nearly 40-fold) over basal levels among mitotic gene convertants selected to arise close to the centromere; no such enhancement is observed among convertants selected to arise relatively far from the centromere. (2) Chromosome loss is primarily associated with proximal allele conversion at the centromere-adjacent site, and many of these convertants are reciprocally recombined in the adjacent proximal interval. (3) Partial aneuploid exceptions provisionally identified as carrying left arm telocentrics have been found. A testable model is proposed suggesting that centromere involvement in genetic recombination may precipitate segregational disfunction leading to mitotic chromosome loss. PMID:324869

  19. The NOXA-MCL1-BIM axis defines lifespan on extended mitotic arrest.

    PubMed

    Haschka, Manuel D; Soratroi, Claudia; Kirschnek, Susanne; Häcker, Georg; Hilbe, Richard; Geley, Stephan; Villunger, Andreas; Fava, Luca L

    2015-01-01

    Cell death on extended mitotic arrest is considered arguably most critical for the efficacy of microtubule-targeting agents (MTAs) in anticancer therapy. While the molecular machinery controlling mitotic arrest on MTA treatment, the spindle assembly checkpoint (SAC), appears well defined, the molecular components executing cell death, as well as factors connecting both networks remain poorly understood. Here we conduct a mini screen exploring systematically the contribution of individual BCL2 family proteins at single cell resolution to death on extended mitotic arrest, and demonstrate that the mitotic phosphorylation of BCL2 and BCLX represent a priming event for apoptosis that is ultimately triggered by NOXA-dependent MCL1 degradation, enabling BIM-dependent cell death. Our findings provide a comprehensive model for the initiation of apoptosis in cells stalled in mitosis and provide a molecular basis for the increased efficacy of combinatorial treatment of cancer cells using MTAs and BH3 mimetics. PMID:25922916

  20. The NOXA–MCL1–BIM axis defines lifespan on extended mitotic arrest

    PubMed Central

    Haschka, Manuel D.; Soratroi, Claudia; Kirschnek, Susanne; Häcker, Georg; Hilbe, Richard; Geley, Stephan; Villunger, Andreas; Fava, Luca L.

    2015-01-01

    Cell death on extended mitotic arrest is considered arguably most critical for the efficacy of microtubule-targeting agents (MTAs) in anticancer therapy. While the molecular machinery controlling mitotic arrest on MTA treatment, the spindle assembly checkpoint (SAC), appears well defined, the molecular components executing cell death, as well as factors connecting both networks remain poorly understood. Here we conduct a mini screen exploring systematically the contribution of individual BCL2 family proteins at single cell resolution to death on extended mitotic arrest, and demonstrate that the mitotic phosphorylation of BCL2 and BCLX represent a priming event for apoptosis that is ultimately triggered by NOXA-dependent MCL1 degradation, enabling BIM-dependent cell death. Our findings provide a comprehensive model for the initiation of apoptosis in cells stalled in mitosis and provide a molecular basis for the increased efficacy of combinatorial treatment of cancer cells using MTAs and BH3 mimetics. PMID:25922916

  1. Inhibition of a Mitotic Motor Protein: Where, How, and Conformational Consequences

    SciTech Connect

    Yan, Youwei; Sardana, Vinod; Xu, Bei; Homnick, Carl; Halczenko, Wasyl; Buser, Carolyn A.; Schaber, Michael; Hartman, George D.; Huber, Hans E.; Kuo, Lawrence C.

    2010-11-16

    We report here the first inhibitor-bound structure of a mitotic motor protein. The 1.9 {angstrom} resolution structure of the motor domain of KSP, bound with the small molecule monastrol and Mg{sup 2+} {center_dot} ADP, reveals that monastrol confers inhibition by 'induced-fitting' onto the protein some 12 {angstrom} away from the catalytic center of the enzyme, resulting in the creation of a previously non-existing binding pocket. The structure provides new insights into the biochemical and mechanical mechanisms of the mitotic motor domain. Inhibition of KSP provides a novel mechanism to arrest mitotic spindle formation, a target of several approved and investigative anti-cancer agents. The structural information gleaned from this novel pocket offers a new angle for the design of anti-mitotic agents.

  2. Physical Description of Mitotic Spindle Orientation During Cell Division

    NASA Astrophysics Data System (ADS)

    Jiménez-Dalmaroni, Andrea; Théry, Manuel; Racine, Victor; Bornens, Michel; Jülicher, Frank

    2009-03-01

    During cell division, the duplicated chromosomes are physically separated by the action of the mitotic spindle. The spindle is a dynamic structure of the cytoskeleton, which consists of two microtubule asters. Its orientation defines the axis along which the cell divides. Recent experiments show that the spindle orientation depends on the spatial distribution of cell adhesion sites. Here we show that the experimentally observed spindle orientation can be understood as the result of the action of cortical force generators acting on the spindle. We assume that the local activity of force generators is controlled by the spatial distribution of cell adhesion sites determined by the particular geometry of the adhesive substrate. We develop a simple physical description of the spindle mechanics, which allows us to calculate the torque acting on the spindle, as well as the energy profile and the angular distribution of spindle orientation. Our model accounts for the preferred spindle orientation, as well as the full shape of the angular distributions of spindle orientation observed in a large variety of pattern geometries. M. Th'ery, A. Jim'enez-Dalmaroni, et al., Nature 447, 493 (2007).

  3. Characterization of a mitotic mutant of durum wheat.

    PubMed

    Klindworth, D L; Williams, N D

    2001-01-01

    An ethyl methanesulfonate-induced mitotic mutant of durum wheat (Triticum turgidum L. var. durum; 2n = 4x = 28) was found. We have characterized the mutant to determine the mechanism of abnormal cell division and to test for temperature effects on abnormal cell division. Stained root-tip meristems and pollen mother cells were studied with brightfield, phase contrast, and immunofluorescence microscopy. Abnormal cells included metaphase cells with a multiple of the normal complement (8x = 56, or 16x = 112), multinucleate cells, 4C, 8C, or 16C mononucleate cells, and cells exhibiting incomplete cytokinesis. The mutant had three classes of pollen mother cells: euploid with normal bivalent pairing, multiploid with bivalent pairing, and multiploid with multivalent pairing. Preprophase bands and spindles were normal in mononucleate cells. Some cells had asymmetrical phragmoplasts and phragmoplast dismantling that produced incomplete cytokinesis. Failure of cytokinesis followed by nuclear fusion were the mechanisms of abnormal cell division. To test for temperature sensitivity of the mutant, seedlings were germinated under six different temperature regimes. As germination temperature increased, the frequency of abnormal cells increased. When the mutant was crossed as the female with durum wheat, 3% of hybrids were hexaploid, indicating that functional-unreduced gametes had formed in megaspores. PMID:11448039

  4. Mitotic chromosome transmission fidelity mutants in Saccharomyces cerevisiae.

    PubMed

    Spencer, F; Gerring, S L; Connelly, C; Hieter, P

    1990-02-01

    We have isolated 136 independent mutations in haploid yeast strains that exhibit decreased chromosome transmission fidelity in mitosis. Eighty-five percent of the mutations are recessive and 15% are partially dominant. Complementation analysis between MATa and MAT alpha isolates identifies 11 chromosome transmission fidelity (CTF) complementation groups, the largest of which is identical to CHL1. For 49 independent mutations, no corresponding allele has been recovered in the opposite mating type. The initial screen monitored the stability of a centromere-linked color marker on a nonessential yeast chromosome fragment; the mitotic inheritance of natural yeast chromosome III is also affected by the ctf mutations. Of the 136 isolates identified, seven were inviable at 37 degrees and five were inviable at 11 degrees. In all cases tested, these temperature conditional lethalities cosegregated with the chromosome instability phenotype. Five additional complementation groups (ctf12 through ctf16) have been defined by complementation analysis of the mutations causing inviability at 37 degrees. Twenty-three of the 136 isolates exhibited growth defects at concentrations of benomyl permissive for the parent strain, and nine appeared to be tolerant of inhibitory levels of benomyl. All of the mutant strains showed normal sensitivity to ultraviolet and gamma-irradiation. Further characterization of these mutant strains will describe the functions of gene products crucial to the successful execution of processes required for aspects of the chromosome cycle that are important for chromosome transmission fidelity in mitosis. PMID:2407610

  5. Growth suppression and mitotic defect induced by JNJ-7706621, an inhibitor of cyclin-dependent kinases and aurora kinases.

    PubMed

    Matsuhashi, A; Ohno, T; Kimura, M; Hara, A; Saio, M; Nagano, A; Kawai, G; Saitou, M; Takigami, I; Yamada, K; Okano, Y; Shimizu, K

    2012-07-01

    Aurora kinases and cyclin-dependent kinases, which play critical roles in the cell cycle and are frequently overexpressed in a variety of tumors, have been suggested as attractive targets for cancer therapy. JNJ-7706621, a recently identified dual inhibitor of these kinases, is reported to induce cell cycle arrest, endoreduplication, and apoptosis. In the present study, we further investigated the molecular mechanisms underlying these effects. The inhibitor arrested various cells at G2 phase at low concentration, and at both G1 and G2 phases at high concentration. JNJ-7706621 did not prevent localization of Aurora A to the spindle poles, but did inhibit other centrosomal proteins such as TOG, Nek2, and TACC3 in early mitotic phase. Similarly, the drug did not prevent localization of Aurora B to the kinetochore, but did inhibit other chromosomal passenger proteins such as Survivin and INCENP. In the cells exposed to JNJ-7706621 after nocodazole release, Aurora B, INCENP, and Survivin became relocated to the peripheral region of chromosomes, but Plk1 and Prc1 were localized on microtubules in later mitotic phase. Treatment of nocodazole-synchronized cells with JNJ-7706621 was able to override mitotic arrest by preventing spindle checkpoint signaling, resulting in failure of chromosome alignment and segregation. Injection of the drug significantly inhibited the growth of TC135 Ewing's sarcoma cells transplanted into athymic mice by cell cycle arrest and apoptosis. JNJ-7706621 is a unique inhibitor regulating cell cycle progression at multiple points, suggesting that it could be useful for cell cycle analysis and therapy of various cancers, including Ewing's sarcoma. PMID:22463590

  6. Stereospecific phosphorylation by the central mitotic kinase Cdk1-cyclin B.

    PubMed

    Etzkorn, Felicia A; Zhao, Song

    2015-04-17

    The cis vs trans conformation, or shape, of phosphoserine-proline (pSer-Pro), a prevalent motif in cell cycle proteins, may play a significant role in regulating mitosis. We demonstrate that Cdk1-cyclin B, the central mitotic kinase, is specific for the trans conformation, not cis, of synthetic, locked Ser-Pro 11-residue peptide substrates, using LC-MSMS detection and sequencing of phosphorylated products. This substrate stereospecificity may contribute an additional level of mitotic regulation. PMID:25603287

  7. The Structure of the Mitotic Spindle and Nucleolus during Mitosis in the Amebo-Flagellate Naegleria

    PubMed Central

    Walsh, Charles J.

    2012-01-01

    Mitosis in the amebo-flagellate Naegleria pringsheimi is acentrosomal and closed (the nuclear membrane does not break down). The large central nucleolus, which occupies about 20% of the nuclear volume, persists throughout the cell cycle. At mitosis, the nucleolus divides and moves to the poles in association with the chromosomes. The structure of the mitotic spindle and its relationship to the nucleolus are unknown. To identify the origin and structure of the mitotic spindle, its relationship to the nucleolus and to further understand the influence of persistent nucleoli on cellular division in acentriolar organisms like Naegleria, three-dimensional reconstructions of the mitotic spindle and nucleolus were carried out using confocal microscopy. Monoclonal antibodies against three different nucleolar regions and α-tubulin were used to image the nucleolus and mitotic spindle. Microtubules were restricted to the nucleolus beginning with the earliest prophase spindle microtubules. Early spindle microtubules were seen as short rods on the surface of the nucleolus. Elongation of the spindle microtubules resulted in a rough cage of microtubules surrounding the nucleolus. At metaphase, the mitotic spindle formed a broad band completely embedded within the nucleolus. The nucleolus separated into two discreet masses connected by a dense band of microtubules as the spindle elongated. At telophase, the distal ends of the mitotic spindle were still completely embedded within the daughter nucleoli. Pixel by pixel comparison of tubulin and nucleolar protein fluorescence showed 70% or more of tubulin co-localized with nucleolar proteins by early prophase. These observations suggest a model in which specific nucleolar binding sites for microtubules allow mitotic spindle formation and attachment. The fact that a significant mass of nucleolar material precedes the chromosomes as the mitotic spindle elongates suggests that spindle elongation drives nucleolar division. PMID:22493714

  8. Quantitative analysis of anthropogenic relief features: automated mapping of charcoal kiln sites from high-resolution ALS data

    NASA Astrophysics Data System (ADS)

    Schneider, Anna; Takla, Melanie; Nicolay, Alexander; Raab, Alexandra; Raab, Thomas

    2014-05-01

    High-resolution digital elevation data from airborne laser scanning (ALS) allow for identification and mapping of so far unknown small-scale relief features that are hidden by forest cover. Especially as a result of historic land use, small anthropogenic landforms can occur, e.g., remains of charcoal kilns on sites that were used for charcoal production or ridge and furrow systems in former farmland areas. Mapping such relief features and analyzing their spatial distribution patterns can help to understand past land-use systems and their effects on landscapes. To efficiently detect and quantify small-scale relief features from high-resolution DEMs for larger areas, (semi-) automated mapping routines are required. In order to describe the number and spatial distribution of historic charcoal kiln sites in the area around Cottbus, Germany, we developed a GIS-based routine for the detection and mapping of kiln remnants from ALS elevation models with a resolution of 1 or 2 meters. The method is based on a template matching algorithm, using a combination of morphometric parameters, and is implemented within ArcGIS. The mapping results could be validated against a comprehensive database of kiln sites and diameters recorded from archaeological excavations in the forefield of the opencast mine Jänschwalde and from manual digitization of kiln remnants from Shaded Relief maps for the Jänschwalder Heide and the Tauersche Forst, north of Cottbus. A considerably high number of charcoal kiln sites could be detected in ALS data, and the diameters of the identified charcoal kilns are remarkable large in the area. For the Jänschwalder Heide, more than 5000 kiln sites in an area of 32 km2 were detected by manual digitization, with 1355 kiln sites that are wider than 12 m. These relatively large kiln sites could be mapped with detection rates that are close to those of manual digitization using the automated mapping routine. Detection quality was improved by the combination of several morphometric parameters used for template matching, as compared to a mapping based on elevation values only. In comparison to manual digitization, a combination of the described detection routine and a manual removal of falsely detected sites can considerably facilitate the mapping and distribution analysis of kiln sites or other small-scale relief features.

  9. Simulating thermal stress features on hot planetary surfaces in vacuum at high temperature facility in the PEL laboratory

    NASA Astrophysics Data System (ADS)

    Maturilli, A.; Ferrari, S.; Helbert, J.; D'Incecco, P.; D'Amore, M.

    2011-12-01

    In the Planetary Emissivity Laboratory (PEL) at the Institute for Planetary Research of the German Aerospace Center (DLR) in Berlin, we set-up a simulation chamber for the spectroscopic investigation of minerals separates under Mercurial conditions. The chamber can be evacuated to 10-4 bar and the target samples heated to 700 K within few minutes, thanks to the innovative inductive heating system. While developing the protocol for the high temperature spectroscopy measurements we discovered interesting "morphologies" on the sample surfaces. The powders are poured into stainless steel cups of 50 mm internal diameter, 8 mm height and 3 mm depth, having a 5 mm thick base (thus leaving 3 mm free space for the minerals), and rim 1 mm thick. We selected several minerals of interest for Mercurial surface composition and for each of them we analyzed various grain size separates, to study the influence of grain dimensions to the process of thermal stressing. We observed that for the smaller grain size separate (0-25 μm) the thermal stress mainly induces large depressions and fractures, while on larger grain sizes (125-250 μm) small depressions and a cratered surface. Our current working hypothesis is that these features are mainly caused by thermal stress induced by a radiatively quickly cooling surface layer covering the much hotter bulk material. Further investigation is ongoing to understand the processes better. The observed morphologies exhibit surprising similarities to features observed at planetary scale size for example on Mercury and even on Venus. Especially the high resolution images provided currently from MESSENGER'S Mercury Dual Imaging System (MDIS) instrument has revealed plains dominated by polygonal fractures whose origin still have to be determined. Our laboratory analogue studies might in the future provide some insight into the processes creating those features

  10. Cross-Talk between AURKA and Plk1 in Mitotic Entry and Spindle Assembly

    PubMed Central

    Asteriti, Italia Anna; De Mattia, Fabiola; Guarguaglini, Giulia

    2015-01-01

    The Aurora kinase A (AURKA) is involved in different aspects of mitotic control, from mitotic entry to cytokinesis. Consistent with its pleiotropic roles, several AURKA interactors are able to modulate its activity, the best characterized being the microtubule-binding protein TPX2, the centrosomal protein Cep192, and Bora. Bora has been described as an essential cofactor of AURKA for phosphorylation-mediated activation of the mitotic kinase polo-like kinase 1 (Plk1) at the G2/M transition. A complex AURKA/Plk1 signaling axis is emerging, with multiple involved actors; recent data suggest that this control network is not restricted to mitotic entry only, but operates throughout mitosis. Here, we integrate available data from the literature to depict the complex interplay between AURKA and Plk1 in G2 and mitosis and how it contributes to their mitotic functions. We will particularly focus on how the activity of specifically localized AURKA/Plk1 pools is modulated in time and space by their reciprocal regulation to ensure the timely and coordinated unfolding of downstream mitotic events. PMID:26779436

  11. MTBP plays a crucial role in mitotic progression and chromosome segregation

    PubMed Central

    Agarwal, N; Tochigi, Y; Adhikari, A S; Cui, S; Cui, Y; Iwakuma, T

    2011-01-01

    Murine double minute 2 (MDM2) binding protein (MTBP) has been implicated in tumor cell proliferation, but the underlying mechanisms remain unclear. The results of MTBP expression analysis during cell cycle progression demonstrated that MTBP protein was rapidly degraded during mitosis. Immunofluorescence studies revealed that a portion of MTBP was localized at the kinetochores during prometaphase. MTBP overexpression delayed mitotic progression from nuclear envelope breakdown (NEB) to anaphase onset and induced abnormal chromosome segregation such as lagging chromosomes, chromosome bridges, and multipolar chromosome segregation. Conversely, MTBP downmodulation caused an abbreviated metaphase and insufficient mitotic arrest, resulting in abnormal chromosome segregation, aneuploidy, decreased cell proliferation, senescence, and cell death, similar to that of Mad2 (mitotic arrest-deficient 2) downmodulation. Furthermore, MTBP downmodulation inhibited the accumulation of Mad1 and Mad2, but not BubR1 (budding uninhibited by benzimidazoles related 1), on the kinetochores, whereas MTBP overexpression inhibited the release of Mad2 from the metaphase kinetochores. These results may imply that MTBP has an important role in recruiting and/or retaining the Mad1/Mad2 complex at the kinetochores during prometaphase, but its degradation is required for silencing the mitotic checkpoint. Together, this study indicates that MTBP has a crucial role in proper mitotic progression and faithful chromosome segregation, providing new insights into regulation of the mitotic checkpoint. PMID:21274008

  12. Genetic and Biochemical Evaluation of the Importance of Cdc6 in Regulating Mitotic ExitD⃞

    PubMed Central

    Archambault, Vincent; Li, Caihong X.; Tackett, Alan J.; Wäsch, Ralph; Chait, Brian T.; Rout, Michael P.; Cross, Frederick R.

    2003-01-01

    We evaluated the hypothesis that the N-terminal region of the replication control protein Cdc6 acts as an inhibitor of cyclin-dependent kinase (Cdk) activity, promoting mitotic exit. Cdc6 accumulation is restricted to the period from mid-cell cycle until the succeeding G1, due to proteolytic control that requires the Cdc6 N-terminal region. During late mitosis, Cdc6 is present at levels comparable with Sic1 and binds specifically to the mitotic cyclin Clb2. Moderate overexpression of Cdc6 promotes viability of CLB2Δdb strains, which otherwise arrest at mitotic exit, and rescue is dependent on the N-terminal putative Cdk-inhibitory domain. These observations support the potential for Cdc6 to inhibit Clb2-Cdk, thus promoting mitotic exit. Consistent with this idea, we observed a cytokinesis defect in cdh1Δ sic1Δ cdc6Δ2–49 triple mutants. However, we were able to construct viable strains, in three different backgrounds, containing neither SIC1 nor the Cdc6 Cdk-inhibitory domain, in contradiction to previous work. We conclude, therefore, that although both Cdc6 and Sic1 have the potential to facilitate mitotic exit by inhibiting Clb2-Cdk, mitotic exit nevertheless does not require any identified stoichiometric inhibitor of Cdk activity. PMID:12960422

  13. Genetic and biochemical evaluation of the importance of Cdc6 in regulating mitotic exit.

    PubMed

    Archambault, Vincent; Li, Caihong X; Tackett, Alan J; Wasch, Ralph; Chait, Brian T; Rout, Michael P; Cross, Frederick R

    2003-11-01

    We evaluated the hypothesis that the N-terminal region of the replication control protein Cdc6 acts as an inhibitor of cyclin-dependent kinase (Cdk) activity, promoting mitotic exit. Cdc6 accumulation is restricted to the period from mid-cell cycle until the succeeding G1, due to proteolytic control that requires the Cdc6 N-terminal region. During late mitosis, Cdc6 is present at levels comparable with Sic1 and binds specifically to the mitotic cyclin Clb2. Moderate overexpression of Cdc6 promotes viability of CLB2Deltadb strains, which otherwise arrest at mitotic exit, and rescue is dependent on the N-terminal putative Cdk-inhibitory domain. These observations support the potential for Cdc6 to inhibit Clb2-Cdk, thus promoting mitotic exit. Consistent with this idea, we observed a cytokinesis defect in cdh1Delta sic1Delta cdc6Delta2-49 triple mutants. However, we were able to construct viable strains, in three different backgrounds, containing neither SIC1 nor the Cdc6 Cdk-inhibitory domain, in contradiction to previous work. We conclude, therefore, that although both Cdc6 and Sic1 have the potential to facilitate mitotic exit by inhibiting Clb2-Cdk, mitotic exit nevertheless does not require any identified stoichiometric inhibitor of Cdk activity. PMID:12960422

  14. Kinesin-13 regulates flagellar, interphase, and mitotic microtubule dynamics in Giardia intestinalis.

    PubMed

    Dawson, Scott C; Sagolla, Meredith S; Mancuso, Joel J; Woessner, David J; House, Susan A; Fritz-Laylin, Lillian; Cande, W Zacheus

    2007-12-01

    Microtubule depolymerization dynamics in the spindle are regulated by kinesin-13, a nonprocessive kinesin motor protein that depolymerizes microtubules at the plus and minus ends. Here we show that a single kinesin-13 homolog regulates flagellar length dynamics, as well as other interphase and mitotic dynamics in Giardia intestinalis, a widespread parasitic diplomonad protist. Both green fluorescent protein-tagged kinesin-13 and EB1 (a plus-end tracking protein) localize to the plus ends of mitotic and interphase microtubules, including a novel localization to the eight flagellar tips, cytoplasmic anterior axonemes, and the median body. The ectopic expression of a kinesin-13 (S280N) rigor mutant construct caused significant elongation of the eight flagella with significant decreases in the median body volume and resulted in mitotic defects. Notably, drugs that disrupt normal interphase and mitotic microtubule dynamics also affected flagellar length in Giardia. Our study extends recent work on interphase and mitotic kinesin-13 functioning in metazoans to include a role in regulating flagellar length dynamics. We suggest that kinesin-13 universally regulates both mitotic and interphase microtubule dynamics in diverse microbial eukaryotes and propose that axonemal microtubules are subject to the same regulation of microtubule dynamics as other dynamic microtubule arrays. Finally, the present study represents the first use of a dominant-negative strategy to disrupt normal protein function in Giardia and provides important insights into giardial microtubule dynamics with relevance to the development of antigiardial compounds that target critical functions of kinesins in the giardial life cycle. PMID:17766466

  15. A Mathematical Model of Mitotic Exit in Budding Yeast: The Role of Polo Kinase

    PubMed Central

    Hancioglu, Baris; Tyson, John J.

    2012-01-01

    Cell cycle progression in eukaryotes is regulated by periodic activation and inactivation of a family of cyclin–dependent kinases (Cdk's). Entry into mitosis requires phosphorylation of many proteins targeted by mitotic Cdk, and exit from mitosis requires proteolysis of mitotic cyclins and dephosphorylation of their targeted proteins. Mitotic exit in budding yeast is known to involve the interplay of mitotic kinases (Cdk and Polo kinases) and phosphatases (Cdc55/PP2A and Cdc14), as well as the action of the anaphase promoting complex (APC) in degrading specific proteins in anaphase and telophase. To understand the intricacies of this mechanism, we propose a mathematical model for the molecular events during mitotic exit in budding yeast. The model captures the dynamics of this network in wild-type yeast cells and 110 mutant strains. The model clarifies the roles of Polo-like kinase (Cdc5) in the Cdc14 early anaphase release pathway and in the G-protein regulated mitotic exit network. PMID:22383977

  16. High performance organic integrated device with ultraviolet photodetective and electroluminescent properties consisting of a charge-transfer-featured naphthalimide derivative

    SciTech Connect

    Wang, Hanyu; Wang, Xu; Yu, Junsheng E-mail: jsyu@uestc.edu.cn; Zhou, Jie; Lu, Zhiyun E-mail: jsyu@uestc.edu.cn

    2014-08-11

    A high performance organic integrated device (OID) with ultraviolet photodetective and electroluminescent (EL) properties was fabricated by using a charge-transfer-featured naphthalimide derivative of 6-(3,5-bis-[9-(4-t-butylphenyl)-9H-carbazol-3-yl]-phenoxy)-2- (4-t-butylphenyl)-benzo[de]isoquinoline-1,3-dione (CzPhONI) as the active layer. The results showed that the OID had a high detectivity of 1.5 × 10{sup 11} Jones at −3 V under the UV-350 nm illumination with an intensity of 0.6 mW/cm{sup 2}, and yielded an exciplex EL light emission with a maximum brightness of 1437 cd/m{sup 2}. Based on the energy band diagram, both the charge transfer feature of CzPhONI and matched energy level alignment were responsible for the dual ultraviolet photodetective and EL functions of OID.

  17. Atmospheric-water absorption features near 2.2 micrometers and their importance in high spectral resolution remote sensing

    NASA Technical Reports Server (NTRS)

    Kruse, F. A.; Clark, R. N.

    1986-01-01

    Selective absorption of electromagnetic radiation by atmospheric gases and water vapor is an accepted fact in terrestrial remote sensing. Until recently, only a general knowledge of atmospheric effects was required for analysis of remote sensing data; however, with the advent of high spectral resolution imaging devices, detailed knowledge of atmospheric absorption bands has become increasingly important for accurate analysis. Detailed study of high spectral resolution aircraft data at the U.S. Geological Survey has disclosed narrow absorption features centered at approximately 2.17 and 2.20 micrometers not caused by surface mineralogy. Published atmospheric transmission spectra and atmospheric spectra derived using the LOWTRAN-5 computer model indicate that these absorption features are probably water vapor. Spectral modeling indicates that the effects of atmospheric absorption in this region are most pronounced in spectrally flat materials with only weak absorption bands. Without correction and detailed knowledge of the atmospheric effects, accurate mapping of surface mineralogy (particularly at low mineral concentrations) is not possible.

  18. New spectral features of stratospheric trace gases identified from high-resolution infrared balloon-borne and laboratory spectra

    NASA Technical Reports Server (NTRS)

    Goldman, A.; Murcray, F. J.; Blatherwick, R. D.; Kosters, J. J.; Murcray, F. H.; Murcray, D. G.; Rinsland, C. P.

    1989-01-01

    A new Michelson-type interferometer system operating in the infrared at very high resolution has been used to record numerous balloon-borne solar absorption spectra of the stratosphere, ground-based solar absorption spectra, and laboratory spectra of molecules of atmospheric interest. In the present work results obtained for several important stratospheric trace gases, HNO3, CIONO2, HO2NO2, NO2, and COF2, in the 8- to 12-micron spectral region are reported. Many new features of these gases have been identified in the stratospheric spectra. Comparison of the new spectra with line-by-line simulations shows that previous spectral line parameters are often inadequate and that new analysis of high-resolution laboratory and atmospheric spectra and improved theoretical calculations will be required for many bands. Preliminary versions of several sets of improved line parameters under development are discussed.

  19. Novel Mad2-targeting miR-493-3p controls mitotic fidelity and cancer cells' sensitivity to paclitaxel.

    PubMed

    Tambe, Mahesh; Pruikkonen, Sofia; Mäki-Jouppila, Jenni; Chen, Ping; Elgaaen, Bente Vilming; Straume, Anne Hege; Huhtinen, Kaisa; Cárpen, Olli; Lønning, Per Eystein; Davidson, Ben; Hautaniemi, Sampsa; Kallio, Marko J

    2016-03-15

    The molecular pathways that contribute to the proliferation and drug response of cancer cells are highly complex and currently insufficiently characterized. We have identified a previously unknown microRNA-based mechanism that provides cancer cells means to stimulate tumorigenesis via increased genomic instability and, at the same time, evade the action of clinically utilized microtubule drugs. We demonstrate miR-493-3p to be a novel negative regulator of mitotic arrest deficient-2 (MAD2), an essential component of the spindle assembly checkpoint that monitors the fidelity of chromosome segregation. The microRNA targets the 3' UTR of Mad2 mRNA thereby preventing translation of the Mad2 protein. In cancer cells, overexpression of miR-493-3p induced a premature mitotic exit that led to increased frequency of aneuploidy and cellular senescence in the progeny cells. Importantly, excess of the miR-493-3p conferred resistance of cancer cells to microtubule drugs. In human neoplasms, miR-493-3p and Mad2 expression alterations correlated with advanced ovarian cancer forms and high miR-493-3p levels were associated with reduced survival of ovarian and breast cancer patients with aggressive tumors, especially in the paclitaxel therapy arm. Our results suggest that intratumoral profiling of miR-493-3p and Mad2 levels can have diagnostic value in predicting the efficacy of taxane chemotherapy. PMID:26943585

  20. The FlyCatwalk: A High-Throughput Feature-Based Sorting System for Artificial Selection in Drosophila

    PubMed Central

    Medici, Vasco; Vonesch, Sibylle Chantal; Fry, Steven N.; Hafen, Ernst

    2015-01-01

    Experimental evolution is a powerful tool for investigating complex traits. Artificial selection can be applied for a specific trait and the resulting phenotypically divergent populations pool-sequenced to identify alleles that occur at substantially different frequencies in the extreme populations. To maximize the proportion of loci that are causal to the phenotype among all enriched loci, population size and number of replicates need to be high. These requirements have, in fact, limited evolution studies in higher organisms, where the time investment required for phenotyping is often prohibitive for large-scale studies. Animal size is a highly multigenic trait that remains poorly understood, and an experimental evolution approach may thus aid in gaining new insights into the genetic basis of this trait. To this end, we developed the FlyCatwalk, a fully automated, high-throughput system to sort live fruit flies (Drosophila melanogaster) based on morphometric traits. With the FlyCatwalk, we can detect gender and quantify body and wing morphology parameters at a four-old higher throughput compared with manual processing. The phenotyping results acquired using the FlyCatwalk correlate well with those obtained using the standard manual procedure. We demonstrate that an automated, high-throughput, feature-based sorting system is able to avoid previous limitations in population size and replicate numbers. Our approach can likewise be applied for a variety of traits and experimental settings that require high-throughput phenotyping. PMID:25556112

  1. Frequency and mitotic heritability of epimutations in Schistosoma mansoni.

    PubMed

    Roquis, David; Rognon, Anne; Chaparro, Cristian; Boissier, Jerome; Arancibia, Nathalie; Cosseau, Celine; Parrinello, Hugues; Grunau, Christoph

    2016-04-01

    Schistosoma mansoni is a parasitic platyhelminth responsible for intestinal bilharzia. It has a complex life cycle, infecting a freshwater snail of the Biomphalaria genus, and then a mammalian host. Schistosoma mansoni adapts rapidly to new (allopatric) strains of its intermediate host. To study the importance of epimutations in this process, we infected sympatric and allopatric mollusc strains with parasite clones. ChIP-Seq was carried out on four histone modifications (H3K4me3, H3K27me3, H3K27ac and H4K20me1) in parallel with genomewide DNA resequencing (i) on parasite larvae shed by the infected snails and (ii) on adult worms that had developed from the larvae. No change in single nucleotide polymorphisms and no mobilization of transposable elements were observed, but 58-105 copy number variations (CNVs) within the parasite clones in different molluscs were detected. We also observed that the allopatric environment induces three types of chromatin structure changes: (i) host-induced changes on larvae epigenomes in 51 regions of the genome that are independent of the parasites' genetic background, (ii) spontaneous changes (not related to experimental condition or genotype of the parasite) at 64 locations and (iii) 64 chromatin structure differences dependent on the parasite genotype. Up to 45% of the spontaneous, but none of the host-induced chromatin structure changes were transmitted to adults. In our model, the environment induces epigenetic changes at specific loci but only spontaneous epimutations are mitotically heritable and have therefore the potential to contribute to transgenerational inheritance. We also show that CNVs are the only source of genetic variation and occur at the same order of magnitude as epimutations. PMID:26826554

  2. High levels of EGFR expression in tumor stroma are associated with aggressive clinical features in epithelial ovarian cancer

    PubMed Central

    Wang, Ke; Li, Dan; Sun, Lu

    2016-01-01

    Purpose The aim of this study was to investigate the clinical significance and biological function of epidermal growth factor receptor (EGFR) expressed in tumor stroma of epithelial ovarian cancer. Methods Immunohistological staining of EGFR was evaluated in 242 patients with epithelial ovarian cancer. The correlations of EGFR expression in tumor stroma with clinicopathological features and with the expression level of Ki-67 were analyzed by SPSS software. Kaplan–Meier analysis and the Cox proportional hazard model were used to analyze the effect of EGFR expression in tumor stroma on the prognosis of patients with epithelial ovarian cancer. Meanwhile, the activities of proliferation and migration of tumor cells were detected when EGFR overexpressed in stroma cells. Results EGFR expression in tumor stroma correlated significantly with clinical stage (χ2=7.002, P=0.008) and distant metastases (χ2=16.59, P<0.001). Furthermore, there was a significantly positive correlation between the level of EGFR expressed in tumor stroma and the level of Ki-67 expressed in tumor cells (χ2=6.120, P=0.013). Patients with high EGFR expression level in tumor stroma showed poor survival (P=0.002). Multivariate analysis showed that high expression of EGFR in tumor stroma was an independent predictor for epithelial ovarian cancer patients (hazard ratio =1.703; 95% confidence interval 1.125–2.578, P=0.012). Furthermore, stroma cells overexpressing EGFR could promote the proliferation and migration of adjacent tumor cells. Conclusion High expression of EGFR in tumor stroma correlates with aggressive clinical features in epithelial ovarian cancer, and is an independent prognostic factor. PMID:26855586

  3. High-Resolution PFPE-based Molding Techniques for Nanofabrication of High-Pattern Density, Sub-20 nm Features: A Fundamental Materials Approach

    SciTech Connect

    Williams, Stuart S.; Retterer, Scott; Lopez, Rene; Ruiz, Ricardo; Samulski, Edward T.; DeSimone, Joseph M.

    2010-04-14

    Several perfluoropolyether (PFPE)-based elastomers for high-resolution replica molding applications are explored. The modulus of the elastomeric materials was increased through synthetic and additive approaches while maintaining relatively low surface tension values (<25 mN/m). Using large area (>4 in.{sup 2}) master templates, we experimentally show the relationship between mold resolution and material properties such as modulus and surface tension for materials used in this study. A composite mold approach was used to form flexible molds out of stiff, high modulus materials that allow for replication of sub-20 nm post structures. Sub-100 nm line grating master templates, formed using e-beam lithography, were used to determine the experimental stability of the molding materials. It was observed that as the feature spacing decreased, high modulus PFPE tetramethacrylate (TMA) composite molds were able to effectively replicate the nanograting structures without cracking or tear-out defects that typically occur with high modulus elastomers.

  4. Knowledge discovery in high-dimensional data: case studies and a user survey for the rank-by-feature framework.

    PubMed

    Seo, Jinwook; Shneiderman, Ben

    2006-01-01

    Knowledge discovery in high-dimensional data is a challenging enterprise, but new visual analytic tools appear to offer users remarkable powers if they are ready to learn new concepts and interfaces. Our three-year effort to develop versions of the Hierarchical Clustering Explorer (HCE) began with building an interactive tool for exploring clustering results. It expanded, based on user needs, to include other potent analytic and visualization tools for multivariate data, especially the rank-by-feature framework. Our own successes using HCE provided some testimonial evidence of its utility, but we felt it necessary to get beyond our subjective impressions. This paper presents an evaluation of the Hierarchical Clustering Explorer (HCE) using three case studies and an e-mail user survey (n = 57) to focus on skill acquisition with the novel concepts and interface for the rank-by-feature framework. Knowledgeable and motivated users in diverse fields provided multiple perspectives that refined our understanding of strengths and weaknesses. A user survey confirmed the benefits of HCE, but gave less guidance about improvements. Both evaluations suggested improved training methods. PMID:16640245

  5. High resolution transmission electron microscope Imaging and first-principles simulations of atomic-scale features in graphene membrane

    NASA Astrophysics Data System (ADS)

    Wang, Wei; Bhandari, Sagar; Yi, Wei; Bell, David; Westervelt, Robert; Kaxiras, Efthimios

    2012-02-01

    Ultra-thin membranes such as graphene[1] are of great importance for basic science and technology applications. Graphene sets the ultimate limit of thinness, demonstrating that a free-standing single atomic layer not only exists but can be extremely stable and strong [2--4]. However, both theory [5, 6] and experiments [3, 7] suggest that the existence of graphene relies on intrinsic ripples that suppress the long-wavelength thermal fluctuations which otherwise spontaneously destroy long range order in a two dimensional system. Here we show direct imaging of the atomic features in graphene including the ripples resolved using monochromatic aberration-corrected transmission electron microscopy (TEM). We compare the images observed in TEM with simulated images based on an accurate first-principles total potential. We show that these atomic scale features can be mapped through accurate first-principles simulations into high resolution TEM contrast. [1] Geim, A. K. & Novoselov, K. S. Nat. Mater. 6, 183-191, (2007). [2] Novoselov, K. S.et al. Science 306, 666-669, (2004). [3] Meyer, J. C. et al. Nature 446, 60-63, (2007). [4] Lee, C., Wei, X. D., Kysar, J. W. & Hone, J. Science 321, 385-388, (2008). [5] Nelson, D. R. & Peliti, L. J Phys-Paris 48, 1085-1092, (1987). [6] Fasolino, A., Los, J. H. & Katsnelson, M. I. Nat. Mater. 6, 858-861, (2007). [7] Meyer, J. C. et al. Solid State Commun. 143, 101-109, (2007).

  6. Spectral features of Mini-Neptunes and EGP orbiting different stars: exploring the effect of high stellar FUV radiation

    NASA Astrophysics Data System (ADS)

    Miguel, Y.; Kaltenegger, L.

    2014-03-01

    Motivated by the growing population of hot exoplanets, we calculated an atmospheric grid for hot mini-Neptune and giant planets, that links astrophysical observable parameters - orbital distance and stellar type - with the atmospheric features expected in transmission and emission spectra. We link a 1D code that calculates the atmospheric thermal structure with a photochemical model that includes disequilibrium chemistry for planets with temperature between 2700K and 700K and explore the effect of empirical model parameters like eddy diffusion coefficients on the results (Miguel & Kaltenegger, 2013). We then use a line by line radiative transfer code to generate the observable spectra. Our models explore the detectable atmospheric features for a wide range of stellar types from F to M for distances between 0.01AU and 0.1 AU. Many main sequence M stars present strong chromospheric activity that produces high-energy radiation. That strongly affects hot exoplanet atmospheres. We use our models to reproduce results for known planets (WASP-12b, CoRoT-2b, XO-1b and HD189733b), model a new planet (HD97658b) and produce a grid for observed planets as well as to inform future observations. Our grid can be applied to current and future observations to characterize exoplanet atmospheres and serves as a reference to interpret atmospheric retrieval analysis results.

  7. Local changes of work function near rough features on Cu surfaces operated under high external electric field

    SciTech Connect

    Djurabekova, Flyura Ruzibaev, Avaz; Parviainen, Stefan; Holmström, Eero; Hakala, Mikko

    2013-12-28

    Metal surfaces operated under high electric fields produce sparks even if they are held in ultra high vacuum. In spite of extensive research on the topic of vacuum arcs, the mystery of vacuum arc origin still remains unresolved. The indications that the sparking rates depend on the material motivate the research on surface response to extremely high external electric fields. In this work by means of density-functional theory calculations we analyze the redistribution of electron density on (100) Cu surfaces due to self-adatoms and in presence of high electric fields from −1 V/nm up to −2 V/nm (−1 to −2 GV/m, respectively). We also calculate the partial charge induced by the external field on a single adatom and a cluster of two adatoms in order to obtain reliable information on charge redistribution on surface atoms, which can serve as a benchmarking quantity for the assessment of the electric field effects on metal surfaces by means of molecular dynamics simulations. Furthermore, we investigate the modifications of work function around rough surface features, such as step edges and self-adatoms.

  8. Identifying Chinese Microblog Users With High Suicide Probability Using Internet-Based Profile and Linguistic Features: Classification Model

    PubMed Central

    Guan, Li; Hao, Bibo; Cheng, Qijin; Yip, Paul SF

    2015-01-01

    Background Traditional offline assessment of suicide probability is time consuming and difficult in convincing at-risk individuals to participate. Identifying individuals with high suicide probability through online social media has an advantage in its efficiency and potential to reach out to hidden individuals, yet little research has been focused on this specific field. Objective The objective of this study was to apply two classification models, Simple Logistic Regression (SLR) and Random Forest (RF), to examine the feasibility and effectiveness of identifying high suicide possibility microblog users in China through profile and linguistic features extracted from Internet-based data. Methods There were nine hundred and nine Chinese microblog users that completed an Internet survey, and those scoring one SD above the mean of the total Suicide Probability Scale (SPS) score, as well as one SD above the mean in each of the four subscale scores in the participant sample were labeled as high-risk individuals, respectively. Profile and linguistic features were fed into two machine learning algorithms (SLR and RF) to train the model that aims to identify high-risk individuals in general suicide probability and in its four dimensions. Models were trained and then tested by 5-fold cross validation; in which both training set and test set were generated under the stratified random sampling rule from the whole sample. There were three classic performance metrics (Precision, Recall, F1 measure) and a specifically defined metric “Screening Efficiency” that were adopted to evaluate model effectiveness. Results Classification performance was generally matched between SLR and RF. Given the best performance of the classification models, we were able to retrieve over 70% of the labeled high-risk individuals in overall suicide probability as well as in the four dimensions. Screening Efficiency of most models varied from 1/4 to 1/2. Precision of the models was generally below 30%. Conclusions Individuals in China with high suicide probability are recognizable by profile and text-based information from microblogs. Although there is still much space to improve the performance of classification models in the future, this study may shed light on preliminary screening of risky individuals via machine learning algorithms, which can work side-by-side with expert scrutiny to increase efficiency in large-scale-surveillance of suicide probability from online social media. PMID:26543921

  9. Breast cancer mitosis detection in histopathological images with spatial feature extraction

    NASA Astrophysics Data System (ADS)

    Albayrak, Abdülkadir; Bilgin, Gökhan

    2013-12-01

    In this work, cellular mitosis detection in histopathological images has been investigated. Mitosis detection is very expensive and time consuming process. Development of digital imaging in pathology has enabled reasonable and effective solution to this problem. Segmentation of digital images provides easier analysis of cell structures in histopathological data. To differentiate normal and mitotic cells in histopathological images, feature extraction step is very crucial step for the system accuracy. A mitotic cell has more distinctive textural dissimilarities than the other normal cells. Hence, it is important to incorporate spatial information in feature extraction or in post-processing steps. As a main part of this study, Haralick texture descriptor has been proposed with different spatial window sizes in RGB and La*b* color spaces. So, spatial dependencies of normal and mitotic cellular pixels can be evaluated within different pixel neighborhoods. Extracted features are compared with various sample sizes by Support Vector Machines using k-fold cross validation method. According to the represented results, it has been shown that separation accuracy on mitotic and non-mitotic cellular pixels gets better with the increasing size of spatial window.

  10. A Signature Inferred from Drosophila Mitotic Genes Predicts Survival of Breast Cancer Patients

    PubMed Central

    Damasco, Christian; Lembo, Antonio; Somma, Maria Patrizia; Gatti, Maurizio; Di Cunto, Ferdinando; Provero, Paolo

    2011-01-01

    Introduction The classification of breast cancer patients into risk groups provides a powerful tool for the identification of patients who will benefit from aggressive systemic therapy. The analysis of microarray data has generated several gene expression signatures that improve diagnosis and allow risk assessment. There is also evidence that cell proliferation-related genes have a high predictive power within these signatures. Methods We thus constructed a gene expression signature (the DM signature) using the human orthologues of 108 Drosophila melanogaster genes required for either the maintenance of chromosome integrity (36 genes) or mitotic division (72 genes). Results The DM signature has minimal overlap with the extant signatures and is highly predictive of survival in 5 large breast cancer datasets. In addition, we show that the DM signature outperforms many widely used breast cancer signatures in predictive power, and performs comparably to other proliferation-based signatures. For most genes of the DM signature, an increased expression is negatively correlated with patient survival. The genes that provide the highest contribution to the predictive power of the DM signature are those involved in cytokinesis. Conclusion This finding highlights cytokinesis as an important marker in breast cancer prognosis and as a possible target for antimitotic therapies. PMID:21386884

  11. Functions of crystallins in and out of lens: Roles in elongated and post-mitotic cells

    PubMed Central

    Slingsby, Christine; Wistow, Graeme J.

    2014-01-01

    The vertebrate lens evolved to collect light and focus it onto the retina. In development, the lens grows through massive elongation of epithelial cells possibly recapitulating the evolutionary origins of the lens. The refractive index of the lens is largely dependent on high concentrations of soluble proteins called crystallins. All vertebrate lenses share a common set of crystallins from two superfamilies (although other lineage specific crystallins exist). The α-crystallins are small heat shock proteins while the β- and γ-crystallins belong to a superfamily that contains structural proteins of uncertain function. The crystallins are expressed at very high levels in lens but are also found at lower levels in other cells, particularly in retina and brain. All these proteins have plausible connections to maintenance of cytoplasmic order and chaperoning of the complex molecular machines involved in the architecture and function of cells, particularly elongated and post-mitotic cells. They may represent a suite of proteins that help maintain homeostasis in such cells that are at risk from stress or from the accumulated insults of aging. PMID:24582830

  12. A novel RING finger protein, human enhancer of invasion 10, alters mitotic progression through regulation of cyclin B levels.

    PubMed

    Toby, Garabet G; Gherraby, Wahiba; Coleman, Thomas R; Golemis, Erica A

    2003-03-01

    The process of cellular morphogenesis is highly conserved in eukaryotes and is dependent upon the function of proteins that are centrally involved in specification of the cell cycle. The human enhancer of invasion clone 10 (HEI10) protein was identified from a HeLa cell library based on its ability to promote yeast agar invasion and filamentation. Through two-hybrid screening, the mitotic cyclin B1 and an E2 ubiquitin-conjugating enzyme were isolated as HEI10-interacting proteins. Mutation of the HEI10 divergent RING finger motif (characteristic of E3 ubiquitin ligases) and Cdc2/cyclin binding and phosphorylation sites alter HEI10-dependent yeast phenotypes, including delay in G(2)/M transition. In vertebrates, the addition of HEI10 inhibits nuclear envelope breakdown and mitotic entry in Xenopus egg extracts. Mechanistically, HEI10 expression reduces cyclin B levels in cycling Xenopus eggs and reduces levels of the cyclin B ortholog Clb2p in yeast. HEI10 is itself a specific in vitro substrate of purified cyclin B/cdc2, with a TPVR motif as primary phosphorylation site. Finally, HEI10 is itself ubiquitinated in egg extracts and is also autoubiquitinated in vitro. These and other points lead to a model in which HEI10 defines a divergent class of E3 ubiquitin ligase, functioning in progression through G(2)/M. PMID:12612082

  13. The Catalytic Subunit of DNA-Dependent Protein Kinase Coordinates with Polo-Like Kinase 1 to Facilitate Mitotic Entry.

    PubMed

    Lee, Kyung-Jong; Shang, Zeng-Fu; Lin, Yu-Fen; Sun, Jingxin; Morotomi-Yano, Keiko; Saha, Debabrata; Chen, Benjamin P C

    2015-04-01

    DNA-dependent protein kinase catalytic subunit (DNA-PKcs) is the key regulator of the non-homologous end joining pathway of DNA double-strand break repair. We have previously reported that DNA-PKcs is required for maintaining chromosomal stability and mitosis progression. Our further investigations reveal that deficiency in DNA-PKcs activity caused a delay in mitotic entry due to dysregulation of cyclin-dependent kinase 1 (Cdk1), the key driving force for cell cycle progression through G2/M transition. Timely activation of Cdk1 requires polo-like kinase 1 (Plk1), which affects modulators of Cdk1. We found that DNA-PKcs physically interacts with Plk1 and could facilitate Plk1 activation both in vitro and in vivo. Further, DNA-PKcs-deficient cells are highly sensitive to Plk1 inhibitor BI2536, suggesting that the coordination between DNA-PKcs and Plk1 is not only crucial to ensure normal cell cycle progression through G2/M phases but also required for cellular resistance to mitotic stress. On the basis of the current study, it is predictable that combined inhibition of DNA-PKcs and Plk1 can be employed in cancer therapy strategy for synthetic lethality. PMID:25925375

  14. Sonic hedgehog-expressing basal cells are general post-mitotic precursors of functional taste receptor cells

    PubMed Central

    Miura, Hirohito; Scott, Jennifer K.; Harada, Shuitsu; Barlow, Linda A.

    2014-01-01

    Background Taste buds contain ~60 elongate cells and several basal cells. Elongate cells comprise three functional taste cell types: I - glial cells, II - bitter/sweet/umami receptor cells, and III - sour detectors. Although taste cells are continuously renewed, lineage relationships among cell types are ill-defined. Basal cells have been proposed as taste bud stem cells, a subset of which express Sonic hedgehog (Shh). However, Shh+ basal cells turnover rapidly suggesting that Shh+ cells are precursors of some or all taste cell types. Results To fate map Shh-expressing cells, mice carrying ShhCreERT2 and a high (CAG-CAT-EGFP) or low (R26RLacZ) efficiency reporter allele were given tamoxifen to activate Cre in Shh+ cells. Using R26RLacZ, lineage-labeled cells occur singly within buds, supporting a post-mitotic state for Shh+ cells. Using either reporter, we show that Shh+ cells differentiate into all three taste cell types, in proportions reflecting cell type ratios in taste buds (I > II > III). Conclusions Shh+ cells are not stem cells, but are post-mitotic, immediate precursors of taste cells. Shh+ cells differentiate into each of the three taste cell types, and the choice of a specific taste cell fate is regulated to maintain the proper ratio within buds. PMID:24590958

  15. The development of wing shape in Lepidoptera: mitotic density, not orientation, is the primary determinant of shape.

    PubMed

    Nijhout, H Frederik; Cinderella, Margaret; Grunert, Laura W

    2014-03-01

    The wings of butterflies and moths develop from imaginal disks whose structure is always congruent with the final adult wing. It is therefore possible to map every point on the imaginal disk to a location on the adult wing throughout ontogeny. We studied the growth patterns of the wings of two distantly related species with very different adult wing shapes, Junonia coenia and Manduca sexta. The shape of the wing disks change throughout their growth phase in a species-specific pattern. We measured mitotic densities and mitotic orientation in successive stages of wing development approximately one cell division apart. Cell proliferation was spatially patterned, and the density of mitoses was highly correlated with local growth. Unlike other systems in which the direction of mitoses has been viewed as the primary determinant of directional growth, we found that in these two species the direction of growth was only weakly correlated with the orientation of mitoses. Directional growth appears to be imposed by a constantly changing spatial pattern of cell division coupled with a weak bias in the orientation of cell division. Because growth and cell division in imaginal disk require ecdysone and insulin signaling, the changing spatial pattern of cell division may due to a changing pattern of expression of receptors or downstream elements in the signaling pathways for one or both of these hormones. Evolution of wing shape comes about by changes in the progression of spatial patterns of cell division. PMID:24617986

  16. Targeting cannabinoid receptor-2 pathway by phenylacetylamide suppresses the proliferation of human myeloma cells through mitotic dysregulation and cytoskeleton disruption.

    PubMed

    Feng, Rentian; Tong, Qin; Xie, Zhaojun; Cheng, Haizi; Wang, Lirong; Lentzsch, Suzanne; Roodman, G David; Xie, Xiang-Qun

    2015-12-01

    Cannabinoid receptor-2 (CB2) is expressed dominantly in the immune system, especially on plasma cells. Cannabinergic ligands with CB2 selectivity emerge as a class of promising agents to treat CB2-expressing malignancies without psychotropic concerns. In this study, we found that CB2 but not CB1 was highly expressed in human multiple myeloma (MM) and primary CD138+ cells. A novel inverse agonist of CB2, phenylacetylamide but not CB1 inverse agonist SR141716, inhibited the proliferation of human MM cells (IC50 : 0.62 ∼ 2.5 μM) mediated by apoptosis induction, but exhibited minor cytotoxic effects on human normal mononuclear cells. CB2 gene silencing or pharmacological antagonism markedly attenuated phenylacetylamide's anti-MM effects. Phenylacetylamide triggered the expression of C/EBP homologous protein at the early treatment stage, followed by death receptor-5 upregulation, caspase activation, and β-actin/tubulin degradation. Cell cycle related protein cdc25C and mitotic regulator Aurora A kinase were inactivated by phenylacetylamide treatment, leading to an increase in the ratio inactive/active cdc2 kinase. As a result, phosphorylation of CDK substrates was decreased, and the MM cell mitotic division was largely blocked by treatment. Importantly, phenylacetylamide could overcome the chemoresistance of MM cells against dexamethasone or melphalan. Thus, targeting CB2 may represent an attractive approach to treat cancers of immune origin. PMID:25640641

  17. Analogues to features and processes of a high-level radioactive waste repository proposed for Yucca Mountain, Nevada

    USGS Publications Warehouse

    Simmons, Ardyth M.; Stuckless, John S.; with a Foreword by Abraham Van Luik, U.S. Department of Energy

    2010-01-01

    Natural analogues are defined for this report as naturally occurring or anthropogenic systems in which processes similar to those expected to occur in a nuclear waste repository are thought to have taken place over time periods of decades to millennia and on spatial scales as much as tens of kilometers. Analogues provide an important temporal and spatial dimension that cannot be tested by laboratory or field-scale experiments. Analogues provide one of the multiple lines of evidence intended to increase confidence in the safe geologic disposal of high-level radioactive waste. Although the work in this report was completed specifically for Yucca Mountain, Nevada, as the proposed geologic repository for high-level radioactive waste under the U.S. Nuclear Waste Policy Act, the applicability of the science, analyses, and interpretations is not limited to a specific site. Natural and anthropogenic analogues have provided and can continue to provide value in understanding features and processes of importance across a wide variety of topics in addressing the challenges of geologic isolation of radioactive waste and also as a contribution to scientific investigations unrelated to waste disposal. Isolation of radioactive waste at a mined geologic repository would be through a combination of natural features and engineered barriers. In this report we examine analogues to many of the various components of the Yucca Mountain system, including the preservation of materials in unsaturated environments, flow of water through unsaturated volcanic tuff, seepage into repository drifts, repository drift stability, stability and alteration of waste forms and components of the engineered barrier system, and transport of radionuclides through unsaturated and saturated rock zones.

  18. Control of high oceanic features and subduction channel on earthquake ruptures along the Chile-Peru subduction zone

    NASA Astrophysics Data System (ADS)

    Contreras-Reyes, Eduardo; Carrizo, Daniel

    2011-05-01

    We discuss the earthquake rupture behavior along the Chile-Peru subduction zone in terms of the buoyancy of the subducting high oceanic features (HOF's), and the effect of the interplay between HOF and subduction channel thickness on the degree of interplate coupling. We show a strong relation between subduction of HOF's and earthquake rupture segments along the Chile-Peru margin, elucidating how these subducting features play a key role in seismic segmentation. Within this context, the extra increase of normal stress at the subduction interface is strongly controlled by the buoyancy of HOF's which is likely caused by crustal thickening and mantle serpentinization beneath hotspot ridges and fracture zones, respectively. Buoyancy of HOF's provide an increase in normal stress estimated to be as high as 10-50 MPa. This significant increase of normal stress will enhance seismic coupling across the subduction interface and hence will affect the seismicity. In particular, several large earthquakes (Mw ≥ 7.5) have occurred in regions characterized by subduction of HOF's including fracture zones (e.g., Nazca, Challenger and Mocha), hotspot ridges (e.g., Nazca, Iquique, and Juan Fernández) and the active Nazca-Antarctic spreading center. For instance, the giant 1960 earthquake (Mw = 9.5) is coincident with the linear projections of the Mocha Fracture Zone and the buoyant Chile Rise, while the active seismic gap of north Chile spatially correlates with the subduction of the Iquique Ridge. Further comparison of rupture characteristics of large underthrusting earthquakes and the locations of subducting features provide evidence that HOF's control earthquake rupture acting as both asperities and barriers. This dual behavior can be partially controlled by the subduction channel thickness. A thick subduction channel smooths the degree of coupling caused by the subducted HOF which allows lateral earthquake rupture propagation. This may explain why the 1960 rupture propagates through six major fracture zones, and ceased near the Mocha Fracture Zone in the north and at the Chile Rise in the south (regions characterized by a thin subduction channel). In addition, the thin subduction channel (north of the Juan Fernández Ridge) reflects a heterogeneous frictional behavior of the subduction interface which appears to be mainly controlled by the subduction of HOF's.

  19. Mitotic Checkpoint Kinase Mps1 Has a Role in Normal Physiology which Impacts Clinical Utility

    PubMed Central

    Martinez, Ricardo; Blasina, Alessandra; Hallin, Jill F.; Hu, Wenyue; Rymer, Isha; Fan, Jeffery; Hoffman, Robert L.; Murphy, Sean; Marx, Matthew; Yanochko, Gina; Trajkovic, Dusko; Dinh, Dac; Timofeevski, Sergei; Zhu, Zhou; Sun, Peiquing; Lappin, Patrick B.; Murray, Brion W.

    2015-01-01

    Cell cycle checkpoint intervention is an effective therapeutic strategy for cancer when applied to patients predisposed to respond and the treatment is well-tolerated. A critical cell cycle process that could be targeted is the mitotic checkpoint (spindle assembly checkpoint) which governs the metaphase-to-anaphase transition and insures proper chromosomal segregation. The mitotic checkpoint kinase Mps1 was selected to explore whether enhancement in genomic instability is a viable therapeutic strategy. The basal-a subset of triple-negative breast cancer was chosen as a model system because it has a higher incidence of chromosomal instability and Mps1 expression is up-regulated. Depletion of Mps1 reduces tumor cell viability relative to normal cells. Highly selective, extremely potent Mps1 kinase inhibitors were created to investigate the roles of Mps1 catalytic activity in tumor cells and normal physiology (PF-7006, PF-3837; Ki<0.5 nM; cellular IC50 2–6 nM). Treatment of tumor cells in vitro with PF-7006 modulates expected Mps1-dependent biology as demonstrated by molecular and phenotypic measures (reduced pHH3-Ser10 levels, shorter duration of mitosis, micro-nucleation, and apoptosis). Tumor-bearing mice treated with PF-7006 exhibit tumor growth inhibition concomitant with pharmacodynamic modulation of a downstream biomarker (pHH3-Ser10). Unfortunately, efficacy only occurs at drug exposures that cause dose-limiting body weight loss, gastrointestinal toxicities, and neutropenia. Mps1 inhibitor toxicities may be mitigated by inducing G1 cell cycle arrest in Rb1-competent cells with the cyclin-dependent kinase-4/6 inhibitor palbociclib. Using an isogenic cellular model system, PF-7006 is shown to be selectively cytotoxic to Rb1-deficient cells relative to Rb1-competent cells (also a measure of kinase selectivity). Human bone marrow cells pretreated with palbociclib have decreased PF-7006-dependent apoptosis relative to cells without palbociclib pretreatment. Collectively, this study raises a concern that single agent therapies inhibiting Mps1 will not be well-tolerated clinically but may be when combined with a selective CDK4/6 drug. PMID:26398286

  20. Mitotic Checkpoint Kinase Mps1 Has a Role in Normal Physiology which Impacts Clinical Utility.

    PubMed

    Martinez, Ricardo; Blasina, Alessandra; Hallin, Jill F; Hu, Wenyue; Rymer, Isha; Fan, Jeffery; Hoffman, Robert L; Murphy, Sean; Marx, Matthew; Yanochko, Gina; Trajkovic, Dusko; Dinh, Dac; Timofeevski, Sergei; Zhu, Zhou; Sun, Peiquing; Lappin, Patrick B; Murray, Brion W

    2015-01-01

    Cell cycle checkpoint intervention is an effective therapeutic strategy for cancer when applied to patients predisposed to respond and the treatment is well-tolerated. A critical cell cycle process that could be targeted is the mitotic checkpoint (spindle assembly checkpoint) which governs the metaphase-to-anaphase transition and insures proper chromosomal segregation. The mitotic checkpoint kinase Mps1 was selected to explore whether enhancement in genomic instability is a viable therapeutic strategy. The basal-a subset of triple-negative breast cancer was chosen as a model system because it has a higher incidence of chromosomal instability and Mps1 expression is up-regulated. Depletion of Mps1 reduces tumor cell viability relative to normal cells. Highly selective, extremely potent Mps1 kinase inhibitors were created to investigate the roles of Mps1 catalytic activity in tumor cells and normal physiology (PF-7006, PF-3837; Ki<0.5 nM; cellular IC50 2-6 nM). Treatment of tumor cells in vitro with PF-7006 modulates expected Mps1-dependent biology as demonstrated by molecular and phenotypic measures (reduced pHH3-Ser10 levels, shorter duration of mitosis, micro-nucleation, and apoptosis). Tumor-bearing mice treated with PF-7006 exhibit tumor growth inhibition concomitant with pharmacodynamic modulation of a downstream biomarker (pHH3-Ser10). Unfortunately, efficacy only occurs at drug exposures that cause dose-limiting body weight loss, gastrointestinal toxicities, and neutropenia. Mps1 inhibitor toxicities may be mitigated by inducing G1 cell cycle arrest in Rb1-competent cells with the cyclin-dependent kinase-4/6 inhibitor palbociclib. Using an isogenic cellular model system, PF-7006 is shown to be selectively cytotoxic to Rb1-deficient cells relative to Rb1-competent cells (also a measure of kinase selectivity). Human bone marrow cells pretreated with palbociclib have decreased PF-7006-dependent apoptosis relative to cells without palbociclib pretreatment. Collectively, this study raises a concern that single agent therapies inhibiting Mps1 will not be well-tolerated clinically but may be when combined with a selective CDK4/6 drug. PMID:26398286

  1. Contribution of High Resolution Microwave and Optical Remote Sensing Observations in Detecting and Monitoring Ocean Coastal Features

    NASA Astrophysics Data System (ADS)

    Gagliardini, D. A.

    Synthetic Aperture Radar SAR satellite sensors have demonstrated their ability to observe ocean features related to dynamical processes Because of the high resolution of available SAR sensors circulation details and small-scale processes can be detected that are not observable by other sensors more frequently used for ocean research such as the NOAA AVHRR and the ORBVIEW2 SeaWiFS In contrast to these LANDSAT-TM thermal and optical channels can be used to observe sea surface temperatures surface layer ocean color upwelled radiance as well as sun glint reflected radiance patterns of surface roughness at a spatial resolution comparable to that of SAR Several examples of TM images obtained in 1997-2003 over the Argentine coastal ocean region where selected from an extensive data set These images were analyzed and compared with a series of SAR images acquired over the same region by the ERS satellites and in some cases near coincident with the TM data This time period allowed the examination of the seasonal cycles as well as interesting episodic events of different ocean processes including currents fronts upwellings algal blooms eddies internal waves and bathymetry signatures Due in situ observations are scarce over this region some of these processes have been documented for first time helping to improve our understanding of some dynamical and biological aspects Therefore it can be concluded that high resolution optical thermal and microwave data have the ability of providing consistent and complementary high-resolution

  2. Realizing one-dimensional quantum and high-frequency transport features in aligned single-walled carbon nanotube ropes

    NASA Astrophysics Data System (ADS)

    Ncube, Siphephile; Chimowa, George; Chiguvare, Zivayi; Bhattacharyya, Somnath

    2014-07-01

    The superiority of the electronic transport properties of single-walled carbon nanotube (SWNT) ropes over SWNT mats is verified from low temperature and frequency-dependent transport. The overall change of resistance versus in nanotube mats shows that 3D variable range hopping is the dominant conduction mechanism within the 2-300 K range. The magneto-resistance (MR) is found to be predominantly negative with a parabolic nature, which can also be described by the hopping model. Although the positive upturn of the MR at low temperatures establishes the contribution from quantum interference, the inherent quantum transport in individual tubes is suppressed at elevated temperatures. Therefore, to minimize multi-channel effects from inter-tube interactions and other defects, two-terminal devices were fabricated from aligned SWNT (extracted from a mat) for low temperature transport as well as high-frequency measurements. In contrast to the mat, the aligned ropes exhibit step-like features in the differential conductance within the 80-300 K temperature range. The effects of plasmon propagation, unique to one dimension, were identified in electronic transport as a non-universal power-law dependence of the differential conductance on temperature and source-drain voltage. The complex impedance showed high power transmission capabilities up to 65 GHz as well as oscillations in the frequency range up to 30 GHz. The measurements suggest that aligned SWNT ropes have a realistic potential for high-speed device applications.

  3. Realizing one-dimensional quantum and high-frequency transport features in aligned single-walled carbon nanotube ropes

    SciTech Connect

    Ncube, Siphephile; Chimowa, George; Chiguvare, Zivayi; Bhattacharyya, Somnath

    2014-07-14

    The superiority of the electronic transport properties of single-walled carbon nanotube (SWNT) ropes over SWNT mats is verified from low temperature and frequency-dependent transport. The overall change of resistance versus in nanotube mats shows that 3D variable range hopping is the dominant conduction mechanism within the 2–300 K range. The magneto-resistance (MR) is found to be predominantly negative with a parabolic nature, which can also be described by the hopping model. Although the positive upturn of the MR at low temperatures establishes the contribution from quantum interference, the inherent quantum transport in individual tubes is suppressed at elevated temperatures. Therefore, to minimize multi-channel effects from inter-tube interactions and other defects, two-terminal devices were fabricated from aligned SWNT (extracted from a mat) for low temperature transport as well as high-frequency measurements. In contrast to the mat, the aligned ropes exhibit step-like features in the differential conductance within the 80–300 K temperature range. The effects of plasmon propagation, unique to one dimension, were identified in electronic transport as a non-universal power-law dependence of the differential conductance on temperature and source-drain voltage. The complex impedance showed high power transmission capabilities up to 65 GHz as well as oscillations in the frequency range up to 30 GHz. The measurements suggest that aligned SWNT ropes have a realistic potential for high-speed device applications.

  4. Comparing the Prognostic Value of PTEN and Akt Expression with the Mitotic Activity Index in Adjuvant Chemotherapy-Treated Node-Negative Breast Cancer patients aged <55 years

    PubMed Central

    Janssen, Emiel A. M.; Søiland, Håvard; Skaland, Ivar; Gudlaugson, Einar; Kjellevold, Kjell H.; Nysted, Arne; Søreide, Jon-Arne; Baak, Jan P. A.

    2007-01-01

    Background: The prognostic value of the PI3K/Akt/mTOR pathway and PTEN in invasive breast cancer (IBC) is controversial. Cell proliferation, especially the Mitotic Activity Index (MAI), is strongly prognostic in lymph node-negative (LNneg) invasive breast cancer. However, its prognostic value has not been compared with the value of Akt and PTEN expression. Material and Methods: Prognostic comparison of Her2Neu, p110alpha (PIK3CA), Akt, mTOR, PTEN, MAI and cell-cycle regulators in 125 LNneg patients aged <55 years with cyclophosphamide, methotrexate, and 5-fluorouracil (CMF)-based adjuvant systemic chemotherapy. Results: Twenty-one (17%) patients developed distant metastases = DMs (median follow-up: 134 months). p110alpha correlated (p = 0.01) with pAkt but only in PTEN-negatives; pAkt correlated (p = 0.02) with mTOR. PTEN-negativity correlated with high MAI, high grade and ER-negativity (p = 0.009). The MAI was the strongest prognosticator (Hazard Ratio = HR = 2.9, p = 0.01). Her2Neu/p110α/Akt/mTOR features have no additional prognostic value to the MAI. PTEN had additional value but only in MAI < 3 (39/125 = 31%; 8% DMs). 19/39 = 49% of the MAI < 3 patients have combined MAI < 3 / PTEN+ with 0% DMs, contrasting 15% DMs in MAI < 3 / PTEN− (p = 0.03). Conclusions: In T1−3N0M0 adjuvant CMF-treated breast cancer patients aged <55 years, MAI was the strongest survival predictor. The PI3K/Akt/mTOR pathway and cell-cycle regulator characteristics had no additional prognostic value, but PTEN has. Patients with combined MAI < 3 & PTEN-positivity had 100% survival. The small subgroup of MAI < 3 patients that died were PTEN-negative. PMID:17429139

  5. Tectonic controls on gravitational deformation in the Mino Mountains, central Japan: a regional sagging-feature mapping in forested mountains using high-resolution airborne LiDAR

    NASA Astrophysics Data System (ADS)

    Kaneda, H.; Kono, T.

    2012-12-01

    Many linear geomorphic features of gravitational origin, known as sagging or sackung, are recognized on and around high mountain ridges worldwide. A complete scanning and mapping of those sagging features in a given region, however, has ever been difficult because classic aerial-photograph examination does not allow detection of small geomorphic features under forest canopies. We here present the first complete distribution map of sagging features in a wide area using high-resolution airborne LiDAR and the elaborate DEM visualization that facilitates mapping and interpretation of small geomorphic features of various morphology and orientations. The target area is the western Mino Mountains, central Japan, where the ~40-km-wide and ~50-km-long area is characterized by relatively monotonous, moderate- to high-relief mountains of 1000-1600 m high and uneven active-fault distribution. The recently acquired 1-m-resolution LiDAR data of the Etsumi Sankei Sabo (Erosion Control) Office cover the entire western Mino Mountains, providing the rare opportunity to examine various controls on large-scale gravitational deformation and mass-wasting in a humid temperate tectonically active region. We produced stereo-paired Red Relief Image Maps to visualize DEMs and carefully mapped sagging features as well as tectonic-geomorphic features. Based on the tentative distribution map, sagging features are generally associated with higher-altitude terrains, indicating that high potential energy is one important factor for the formation of sagging features. The sagging features also appear to be concentrated around active faults, in particular around fault tips and in fault step-over areas, which may suggest strong tectonic controls on gravitational deformation. In contrast, lithology does not seem to be a major factor to control gravitational deformation in the studied area.

  6. PP1 initiates the dephosphorylation of MASTL, triggering mitotic exit and bistability in human cells

    PubMed Central

    Rogers, Samuel; Fey, Dirk; McCloy, Rachael A.; Parker, Benjamin L.; Mitchell, Nicholas J.; Payne, Richard J.; Daly, Roger J.; James, David E.; Caldon, C. Elizabeth; Watkins, D. Neil; Croucher, David R.; Burgess, Andrew

    2016-01-01

    ABSTRACT Entry into mitosis is driven by the phosphorylation of thousands of substrates, under the master control of Cdk1. During entry into mitosis, Cdk1, in collaboration with MASTL kinase, represses the activity of the major mitotic protein phosphatases, PP1 and PP2A, thereby ensuring mitotic substrates remain phosphorylated. For cells to complete and exit mitosis, these phosphorylation events must be removed, and hence, phosphatase activity must be reactivated. This reactivation of phosphatase activity presumably requires the inhibition of MASTL; however, it is not currently understood what deactivates MASTL and how this is achieved. In this study, we identified that PP1 is associated with, and capable of partially dephosphorylating and deactivating, MASTL during mitotic exit. Using mathematical modelling, we were able to confirm that deactivation of MASTL is essential for mitotic exit. Furthermore, small decreases in Cdk1 activity during metaphase are sufficient to initiate the reactivation of PP1, which in turn partially deactivates MASTL to release inhibition of PP2A and, hence, create a feedback loop. This feedback loop drives complete deactivation of MASTL, ensuring a strong switch-like activation of phosphatase activity during mitotic exit. PMID:26872783

  7. Aurora B prevents delayed DNA replication and premature mitotic exit by repressing p21Cip1

    PubMed Central

    Trakala, Marianna; Fernndez-Miranda, Gonzalo; Prez de Castro, Ignacio; Heeschen, Christopher; Malumbres, Marcos

    2013-01-01

    Aurora kinase B is a critical component of the chromosomal passenger complex, which is involved in the regulation of microtubule-kinetochore attachments and cytokinesis. By using conditional knockout cells and chemical inhibition, we show here that inactivation of Aurora B results in delayed G1/S transition and premature mitotic exit. Aurora B deficiency results in delayed DNA replication in cultured fibroblasts as well as liver cells after hepatectomy. This is accompanied by increased transcription of the cell cycle inhibitor p21Cip1. Lack of Aurora B does not prevent mitotic entry but results in a premature exit from prometaphase in the presence of increased p21Cip1-Cdk1 inactive complexes. Aurora B-null cells display reduced degradation of cyclin B1, suggesting the presence of phenomenon known as adaptation to the mitotic checkpoint, previously described in yeast. Elimination of p21Cip1 rescues Cdk1 activity and prevents premature mitotic exit in Aurora B-deficient cells. These results suggest that Aurora B represses p21Cip1, preventing delayed DNA replication, Cdk inhibition and premature mitotic exit. The upregulation of p21Cip1 observed after inhibition of Aurora B may have important implications in cell cycle progression, tetraploidy, senescence or cancer therapy. PMID:23428904

  8. Heat shock protein inhibitors, 17-DMAG and KNK437, enhance arsenic trioxide-induced mitotic apoptosis

    SciTech Connect

    Wu Yichen; Yen Wenyen; Lee, T.-C. Yih, L.-H.

    2009-04-15

    Arsenic trioxide (ATO) has recently emerged as a promising therapeutic agent in leukemia because of its ability to induce apoptosis. However, there is no sufficient evidence to support its therapeutic use for other types of cancers. In this study, we investigated if, and how, 17-dimethylaminoethylamino-17-demethoxy-geldanamycin (17-DMAG), an antagonist of heat shock protein 90 (HSP90), and KNK437, a HSP synthesis inhibitor, potentiated the cytotoxic effect of ATO. Our results showed that cotreatment with ATO and either 17-DMAG or KNK437 significantly increased ATO-induced cell death and apoptosis. siRNA-mediated attenuation of the expression of the inducible isoform of HSP70 (HSP70i) or HSP90{alpha}/{beta} also enhanced ATO-induced apoptosis. In addition, cotreatment with ATO and 17-DMAG or KNK437 significantly increased ATO-induced mitotic arrest and ATO-induced BUBR1 phosphorylation and PDS1 accumulation. Cotreatment also significantly increased the percentage of mitotic cells with abnormal mitotic spindles and promoted metaphase arrest as compared to ATO treatment alone. These results indicated that 17-DMAG or KNK437 may enhance ATO cytotoxicity by potentiating mitotic arrest and mitotic apoptosis possibly through increased activation of the spindle checkpoint.

  9. Mechanisms and Regulation of the Mitotic Inheritance of the Golgi Complex.

    PubMed

    Valente, Carmen; Colanzi, Antonino

    2015-01-01

    In mammalian cells, the Golgi complex is structured in the form of a continuous membranous system composed of stacks connected by tubular bridges: the "Golgi ribbon." At the onset of mitosis, the Golgi complex undergoes a multi-step fragmentation process that is required for its correct partition into the dividing cells. Importantly, inhibition of Golgi disassembly results in cell-cycle arrest at the G2 stage, which indicates that accurate inheritance of the Golgi complex is monitored by a "Golgi mitotic checkpoint." Moreover, mitotic Golgi disassembly correlates with the release of a set of Golgi-localized proteins that acquire specific functions during mitosis, such as mitotic spindle formation and regulation of the spindle checkpoint. Most of these events are regulated by small GTPases of the Arf and Rab families. Here, we review recent studies that are revealing the fundamental mechanisms, the molecular players, and the biological significance of mitotic inheritance of the Golgi complex in mammalian cells. We also briefly comment on how Golgi partitioning is coordinated with mitotic progression. PMID:26734607

  10. The Developmental Genetics of Hybrid Inviability: A Mitotic Defect in Drosophila Hybrids

    PubMed Central

    Orr, H. A.; Madden, L. D.; Coyne, J. A.; Goodwin, R.; Hawley, R. S.

    1997-01-01

    We report studies of the developmental basis of hybrid inviability in the Drosophila melanogaster complex. The pathology of these hybrids closely resembles that of mitotic mutants in D. melanogaster. We use mosaic and cytological analyses to show that hybrid male inviability is associated with, and probably caused by, a defect in mitotic cell division. In the mosaic study, we find that male clones produced in otherwise female hybrids are not cell lethal but are very small, probably reflecting defects in mitotic proliferation. Cytological inspection of larval neuroblasts reveals a profound mitotic defect in hybrids: chromosomes show a near-complete failure to condense even after 2 hr of incubation in colchicine. Both the defect in clonal proliferation and in chromatin condensation are rescued by mutations known to rescue normally inviable hybrid males. We present a simple model in which hybrid inviability is partly or entirely caused by a mitotic defect; this defect is, in turn, caused by an interaction between the Hybrid male rescue (Hmr) locus of D. melanogaster and autosomal gene(s) from D. melanogaster's sister species. PMID:9093855

  11. Mechanisms and Regulation of the Mitotic Inheritance of the Golgi Complex

    PubMed Central

    Valente, Carmen; Colanzi, Antonino

    2015-01-01

    In mammalian cells, the Golgi complex is structured in the form of a continuous membranous system composed of stacks connected by tubular bridges: the “Golgi ribbon.” At the onset of mitosis, the Golgi complex undergoes a multi-step fragmentation process that is required for its correct partition into the dividing cells. Importantly, inhibition of Golgi disassembly results in cell-cycle arrest at the G2 stage, which indicates that accurate inheritance of the Golgi complex is monitored by a “Golgi mitotic checkpoint.” Moreover, mitotic Golgi disassembly correlates with the release of a set of Golgi-localized proteins that acquire specific functions during mitosis, such as