WorldWideScience

Sample records for formins drfs daam1

  1. Daam1 regulates fascin for actin assembly in mouse oocyte meiosis.

    Science.gov (United States)

    Lu, Yujie; Zhang, Yu; Pan, Meng-Hao; Kim, Nam-Hyung; Sun, Shao-Chen; Cui, Xiang-Shun

    2017-07-18

    As a formin protein, Daam1 (Dishevelled-associated activator of morphogenesis 1) is reported to regulate series of cell processes like endocytosis, cell morphology and migration via its effects on actin assembly in mitosis. However, whether Daam1 plays roles in female meiosis remains uncertain. In this study, we investigated the expression and functions of Daam1 during mouse oocyte meiosis. Our results indicated that Daam1 localized at the cortex of oocytes, which was similar with actin filaments. After Daam1 morpholino (MO) microinjection, the expression of Daam1 significantly decreased, which resulted in the failure of oocyte polar body extrusion. These results might be due to the defects of actin assembly, since the decreased fluorescence intensity of actin filaments in oocyte cortex and cytoplasm were observed. However, Daam1 knockdown seemed not to affect the meiotic spindle movement. In addition, we found that fascin might be the down effector of Daam1, since the protein expression of fascin decreased after Daam1 knockdown. Thus, our data suggested that Daam1 affected actin assembly during oocyte meiotic division via the regulation of fascin expression.

  2. Formin' actin in the nucleus.

    Science.gov (United States)

    Baarlink, Christian; Grosse, Robert

    2014-01-01

    Many if not most proteins can, under certain conditions, change cellular compartments, such as, for example, shuttling from the cytoplasm to the nucleus. Thus, many proteins may exert functions in various and very different subcellular locations, depending on the signaling context. A large amount of actin regulatory proteins has been detected in the mammalian cell nucleus, although their potential roles are much debated and are just beginning to emerge. Recently, members of the formin family of actin nucleators were also reported to dynamically localize to the nuclear environment. Here we discuss our findings that specific diaphanous-related formins can promote nuclear actin assembly in a signal-dependent manner.

  3. Initiation of DNA replication requires actin dynamics and formin activity.

    Science.gov (United States)

    Parisis, Nikolaos; Krasinska, Liliana; Harker, Bethany; Urbach, Serge; Rossignol, Michel; Camasses, Alain; Dewar, James; Morin, Nathalie; Fisher, Daniel

    2017-11-02

    Nuclear actin regulates transcriptional programmes in a manner dependent on its levels and polymerisation state. This dynamics is determined by the balance of nucleocytoplasmic shuttling, formin- and redox-dependent filament polymerisation. Here, using Xenopus egg extracts and human somatic cells, we show that actin dynamics and formins are essential for DNA replication. In proliferating cells, formin inhibition abolishes nuclear transport and initiation of DNA replication, as well as general transcription. In replicating nuclei from transcriptionally silent Xenopus egg extracts, we identified numerous actin regulators, and disruption of actin dynamics abrogates nuclear transport, preventing NLS (nuclear localisation signal)-cargo release from RanGTP-importin complexes. Nuclear formin activity is further required to promote loading of cyclin-dependent kinase (CDK) and proliferating cell nuclear antigen (PCNA) onto chromatin, as well as initiation and elongation of DNA replication. Therefore, actin dynamics and formins control DNA replication by multiple direct and indirect mechanisms. © 2017 The Authors.

  4. FMNL formins boost lamellipodial force generation

    DEFF Research Database (Denmark)

    Kage, Frieda; Winterhoff, Moritz; Dimchev, Vanessa

    2017-01-01

    , without changing patterns of Arp2/3 complex incorporation. Strikingly, in melanoma cells, FMNL2/3 gene inactivation almost completely abolishes protrusion forces exerted by lamellipodia and modifies their ultrastructural organization. Consistently, CRISPR/Cas-mediated depletion of FMNL2/3 in fibroblasts...... reduces both migration and capability of cells to move against viscous media. Together, we conclude that force generation in lamellipodia strongly depends on FMNL formin activity, operating in addition to Arp2/3 complex-dependent filament branching....

  5. Formins: Linking Cytoskeleton and Endomembranes in Plant Cells

    Czech Academy of Sciences Publication Activity Database

    Cvrčková, F.; Oulehlová, Denisa; Žárský, Viktor

    2015-01-01

    Roč. 16, č. 1 (2015), s. 1-18 E-ISSN 1422-0067 Institutional support: RVO:61389030 Keywords : formin * actin * microtubules Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.257, year: 2015

  6. Revisiting the Phylogeny of the Animal Formins: Two New Subtypes, Relationships with Multiple Wing Hairs Proteins, and a Lost Human Formin.

    Science.gov (United States)

    Pruyne, David

    2016-01-01

    Formins are a widespread family of eukaryotic cytoskeleton-organizing proteins. Many species encode multiple formin isoforms, and for animals, much of this reflects the presence of multiple conserved subtypes. Earlier phylogenetic analyses identified seven major formin subtypes in animals (DAAM, DIAPH, FHOD, FMN, FMNL, INF, and GRID2IP/delphilin), but left a handful of formins, particularly from nematodes, unassigned. In this new analysis drawing from genomic data from a wider range of taxa, nine formin subtypes are identified that encompass all the animal formins analyzed here. Included in this analysis are Multiple Wing Hairs proteins (MWH), which bear homology to formin N-terminal domains. Originally identified in Drosophila melanogaster and other arthropods, MWH-related proteins are also identified here in some nematodes (including Caenorhabditis elegans), and are shown to be related to a novel MWH-related formin (MWHF) subtype. One surprising result of this work is the discovery that a family of pleckstrin homology domain-containing formins (PHCFs) is represented in many vertebrates, but is strikingly absent from placental mammals. Consistent with a relatively recent loss of this formin, the human genome retains fragments of a defunct homologous formin gene.

  7. The Diaphanous-related Formin FHOD1 associates with ROCK1 and promotes Src-dependent plasma membrane blebbing.

    Science.gov (United States)

    Hannemann, Sebastian; Madrid, Ricardo; Stastna, Jana; Kitzing, Thomas; Gasteier, Judith; Schönichen, André; Bouchet, Jerome; Jimenez, Alberto; Geyer, Matthias; Grosse, Robert; Benichou, Serge; Fackler, Oliver T

    2008-10-10

    Diaphanous-related formins (DRFs) mediate GTPase-triggered actin rearrangements to regulate central cellular processes, such as cell motility and cytokinesis. The DRF FHOD1 interacts with the Rho-GTPase Rac1 and mediates formation of actin stress fibers in its deregulated form; the physiologically relevant activities and molecular mechanisms of endogenous FHOD1, however, are still unknown. Here we report that FHOD1 physically associates via the N-terminal part of its FH2 domain with the central domain of ROCK1. Although FHOD1 does not affect the kinase activity of ROCK1, the DRF is an efficient substrate for phosphorylation by ROCK1. Co-expression of FHOD1 and ROCK1 results in the generation of nonapoptotic plasma membrane (PM) blebs, to which the DRF is efficiently recruited. Blebbing induced by FHOD1 and ROCK1 depends on F-actin integrity, the Rho-ROCK cascade, and Src activity and is reminiscent of the recently described PM blebs triggered by expression of Src homology 4 (SH4) domain PM targeting signals. Consistently, endogenous FHOD1 is required in SH4 domain expressing cells for efficient PM blebbing and rounded cell morphology in two-dimensional cultures or three-dimensional matrices, respectively. Efficient association of FHOD1 with ROCK1, as well as recruitment of the DRF to blebs, depends on Src activity, suggesting that the functional interaction between both proteins is regulated by Src. These results define a role for endogenous FHOD1 in SH4 domain-induced blebbing and suggest that its activity is regulated by ROCK1 in a Src-dependent manner.

  8. Formin homology 2 domains occur in multiple contexts in angiosperms

    Czech Academy of Sciences Publication Activity Database

    Cvrčková, F.; Novotný, M.; Pícková, Denisa; Žárský, Viktor

    2004-01-01

    Roč. 5, č. 44 (2004), s. 1-18 ISSN 1471-2164 R&D Projects: GA AV ČR KSK5052113 Keywords : Formin * angiosperms * Arabidopsis Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.250, year: 2004

  9. [At the plant side of formins--organizers of the actin cytoskeleton].

    Science.gov (United States)

    Maruniewicz, Michalina; Kasprowicz, Anna; Wojtaszek, Przemysław

    2009-01-01

    Rearrangements of actin cytoskeleton enable proper functioning of the cells under normal conditions, and also cellular adaptations to changes in the direct surroundings. Formins are actin binding proteins, responsible for actin nucleation and further elongation of microfilaments. The distinguishing feature of formins is the presence of conserved FH2 (formin homology domain 2) domain, as well as other domains typical for distinct formin classes. In animal cells formins are involved in cytokinesis and determination and maintenance of the cell shape and polarity, but also in the formation of filopodia, endocytosis and many other processes. The presence of proteins from the formin family in plant cells, and their involvement in the tip growth and cytokinesis, has been determined only recently. As the functional organization of plant and animal cells is different, one can assume that the range of putative functions of plant formins might also be diverse. One of such proposed functions for formins in plants is the role of linker protein within WMC continuum (cell wall-plasma membrane-cytoskeleton). Unfortunately, for that moment the state of knowledge about plant formins in comparison with animal or fungal ones is much poorer.

  10. Centrosome polarization in T cells: a task for formins

    Directory of Open Access Journals (Sweden)

    Laura eAndrés-Delgado

    2013-07-01

    Full Text Available T-cell antigen receptor (TCR engagement triggers the rapid reorientation of the centrosome, which is associated with the secretory machinery, towards the immunological synapse (IS for polarized protein trafficking. Recent evidence indicates that upon TCR triggering the INF2 formin, together with the formins DIA1 and FMNL1, promotes the formation of a specialized array of stable detyrosinated MTs that breaks the symmetrical organization of the T-cell microtubule (MT cytoskeleton. The detyrosinated MT array and TCR-induced tyrosine phosphorylation should coincide for centrosome polarization. We propose that the pushing forces produced by the detyrosinated MT array, which modify the position of the centrosome, in concert with Src kinase dependent TCR signaling, which provide the reference frame with respect to which the centrosome reorients, result in the repositioning of the centrosome to the IS.

  11. Small GTPases and formins in mammalian oocyte maturation: cytoskeletal organizers.

    Science.gov (United States)

    Kwon, Sojung; Lim, Hyunjung J

    2011-03-01

    The maturation process of mammalian oocytes accompanies an extensive rearrangement of the cytoskeleton and associated proteins. As this process requires a delicate interplay between the cytoskeleton and its regulators, it is often targeted by various external and internal adversaries that affect the congression and/or segregation of chromosomes. Asymmetric cell division in oocytes also requires specific regulators of the cytoskeleton, including formin-2 and small GTPases. Recent literature providing clues regarding how actin filaments and microtubules interact during spindle migration in mouse oocytes are highlighted in this review.

  12. FH3, a domain found in formins, targets the fission yeast formin Fus1 to the projection tip during conjugation

    DEFF Research Database (Denmark)

    Petersen, J; Nielsen, O; Egel, R

    1998-01-01

    of the late G2 cells in a vegetatively growing population. Expression of both FH3-GFP fusions also affected cytokinesis. Overexpression of the spindle pole body component Sad1 altered the distribution of both Sad1 and the FH3-GFP domain. Together these data suggest that proteins at multiple sites can interact...... is required for conjugation, and is localized to the projection tip in cells of mating pairs. We replaced genomic fus1+ with green fluorescent protein (GFP)- tagged versions that lacked either the FH1, FH2, or FH3 domain. Deletion of any FH domain essentially abolished mating. FH3, but neither FH1 nor FH2......, was required for Fus1 localization. An FH3 domain-GFP fusion protein localized to the projection tips of mating pairs. Thus, the FH3 domain alone can direct protein localization. The FH3 domains of both Fus1 and the S. pombe cytokinesis formin Cdc12 were able to localize GFP to the spindle pole body in half...

  13. Differential Toxicity of mDia Formin-Directed Functional Agonists and Antagonists in Developing Zebrafish

    Directory of Open Access Journals (Sweden)

    Hunter LeCorgne

    2018-04-01

    Full Text Available The mammalian Diaphanous-related (mDia formins are cytoskeletal regulators that assemble and, in some cases, bundle filamentous actin (F-actin, as well as stabilize microtubules. The development of small molecule antagonists and agonists that interrogate mDia formin function has allowed us to investigate the roles of formins in disease states. A small molecule inhibitor of FH2 domain (SMIFH2 inhibits mDia-dependent actin dynamics and abrogates tumor cell migration and cell division in vitro and ex vivo tissue explants. mDia formin activation with small molecule intramimics IMM01/02 and mDia2-DAD peptides inhibited glioblastoma motility and invasion in vitro and ex vivo rat brain slices. However, SMIFH2, IMMs, and mDia2 DAD efficacy in vivo remains largely unexplored and potential toxicity across a range of developmental phenotypes has not been thoroughly characterized. In this study, we performed an in vivo screen of early life-stage toxicity in Danio rerio zebrafish embryos 2 days post-fertilization (dpf in response to SMIFH2, IMM01/02, and mDia2 DAD. SMIFH2 at concentrations ≥5–10 μM induced significant defects in developing zebrafish, including shorter body lengths, tail curvature and defective tail cellularity, craniofacial malformations, pericardial edema, absent and/or compromised vasculature function and flow, depressed heart rates and increased mortality. Conversely, IMM and mDia2 DAD peptides were minimally toxic at concentrations up to 10–20 and 50 μM, respectively. SMIFH2's therapeutic potential may therefore be limited by its substantial in vivo toxicity at functional concentrations. mDia formin agonism with IMMs and mDia2 DADs may therefore be a more effective and less toxic anti-invasive therapeutic approach.

  14. A comparative sequence analysis reveals a common GBD/FH3-FH1-FH2-DAD architecture in formins from Dictyostelium, fungi and metazoa

    Directory of Open Access Journals (Sweden)

    Uyeda Taro QP

    2005-03-01

    Full Text Available Abstract Background Formins are multidomain proteins defined by a conserved FH2 (formin homology 2 domain with actin nucleation activity preceded by a proline-rich FH1 (formin homology 1 domain. Formins act as profilin-modulated processive actin nucleators conserved throughout a wide range of eukaryotes. Results We present a detailed sequence analysis of the 10 formins (ForA to J identified in the genome of the social amoeba Dictyostelium discoideum. With the exception of ForI and ForC all other formins conform to the domain structure GBD/FH3-FH1-FH2-DAD, where DAD is the Diaphanous autoinhibition domain and GBD/FH3 is the Rho GTPase-binding domain/formin homology 3 domain that we propose to represent a single domain. ForC lacks a FH1 domain, ForI lacks recognizable GBD/FH3 and DAD domains and ForA, E and J have additional unique domains. To establish the relationship between formins of Dictyostelium and other organisms we constructed a phylogenetic tree based on the alignment of FH2 domains. Real-time PCR was used to study the expression pattern of formin genes. Expression of forC, D, I and J increased during transition to multi-cellular stages, while the rest of genes displayed less marked developmental variations. During sexual development, expression of forH and forI displayed a significant increase in fusion competent cells. Conclusion Our analysis allows some preliminary insight into the functionality of Dictyostelium formins: all isoforms might display actin nucleation activity and, with the exception of ForI, might also be susceptible to autoinhibition and to regulation by Rho GTPases. The architecture GBD/FH3-FH1-FH2-DAD appears common to almost all Dictyostelium, fungal and metazoan formins, for which we propose the denomination of conventional formins, and implies a common regulatory mechanism.

  15. A comparative sequence analysis reveals a common GBD/FH3-FH1-FH2-DAD architecture in formins from Dictyostelium, fungi and metazoa.

    Science.gov (United States)

    Rivero, Francisco; Muramoto, Tetsuya; Meyer, Ann-Kathrin; Urushihara, Hideko; Uyeda, Taro Q P; Kitayama, Chikako

    2005-03-01

    Formins are multidomain proteins defined by a conserved FH2 (formin homology 2) domain with actin nucleation activity preceded by a proline-rich FH1 (formin homology 1) domain. Formins act as profilin-modulated processive actin nucleators conserved throughout a wide range of eukaryotes. We present a detailed sequence analysis of the 10 formins (ForA to J) identified in the genome of the social amoeba Dictyostelium discoideum. With the exception of ForI and ForC all other formins conform to the domain structure GBD/FH3-FH1-FH2-DAD, where DAD is the Diaphanous autoinhibition domain and GBD/FH3 is the Rho GTPase-binding domain/formin homology 3 domain that we propose to represent a single domain. ForC lacks a FH1 domain, ForI lacks recognizable GBD/FH3 and DAD domains and ForA, E and J have additional unique domains. To establish the relationship between formins of Dictyostelium and other organisms we constructed a phylogenetic tree based on the alignment of FH2 domains. Real-time PCR was used to study the expression pattern of formin genes. Expression of forC, D, I and J increased during transition to multi-cellular stages, while the rest of genes displayed less marked developmental variations. During sexual development, expression of forH and forI displayed a significant increase in fusion competent cells. Our analysis allows some preliminary insight into the functionality of Dictyostelium formins: all isoforms might display actin nucleation activity and, with the exception of ForI, might also be susceptible to autoinhibition and to regulation by Rho GTPases. The architecture GBD/FH3-FH1-FH2-DAD appears common to almost all Dictyostelium, fungal and metazoan formins, for which we propose the denomination of conventional formins, and implies a common regulatory mechanism.

  16. Arabidopsis FH1 Formin Affects Cotyledon Pavement Cell Shape by Modulating Cytoskeleton Dynamics.

    Science.gov (United States)

    Rosero, Amparo; Oulehlová, Denisa; Stillerová, Lenka; Schiebertová, Petra; Grunt, Michal; Žárský, Viktor; Cvrčková, Fatima

    2016-03-01

    Plant cell morphogenesis involves concerted rearrangements of microtubules and actin microfilaments. We previously reported that FH1, the main Arabidopsis thaliana housekeeping Class I membrane-anchored formin, contributes to actin dynamics and microtubule stability in rhizodermis cells. Here we examine the effects of mutations affecting FH1 (At3g25500) on cell morphogenesis and above-ground organ development in seedlings, as well as on cytoskeletal organization and dynamics, using a combination of confocal and variable angle epifluorescence microscopy with a pharmacological approach. Homozygous fh1 mutants exhibited cotyledon epinasty and had larger cotyledon pavement cells with more pronounced lobes than the wild type. The pavement cell shape alterations were enhanced by expression of the fluorescent microtubule marker GFP-microtubule-associated protein 4 (MAP4). Mutant cotyledon pavement cells exhibited reduced density and increased stability of microfilament bundles, as well as enhanced dynamics of microtubules. Analogous results were also obtained upon treatments with the formin inhibitor SMIFH2 (small molecule inhibitor of formin homology 2 domains). Pavement cell shape in wild-type (wt) and fh1 plants in some situations exhibited a differential response towards anti-cytoskeletal drugs, especially the microtubule disruptor oryzalin. Our observations indicate that FH1 participates in the control of microtubule dynamics, possibly via its effects on actin, subsequently influencing cell morphogenesis and macroscopic organ development. © The Author 2016. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  17. FHOD1 formin is upregulated in melanomas and modifies proliferation and tumor growth

    International Nuclear Information System (INIS)

    Peippo, Minna; Gardberg, Maria; Lamminen, Tarja; Kaipio, Katja; Carpén, Olli; Heuser, Vanina D.

    2017-01-01

    The functional properties of actin-regulating formin proteins are diverse and in many cases cell-type specific. FHOD1, a formin expressed predominantly in cells of mesenchymal lineage, bundles actin filaments and participates in maintenance of cell shape, migration and cellular protrusions. FHOD1 participates in cancer-associated epithelial to mesenchymal transition (EMT) in oral squamous cell carcinoma and breast cancer. The role of FHOD1 in melanomas has not been characterized. Here, we show that FHOD1 expression is typically strong in cutaneous melanomas and cultured melanoma cells while the expression is low or absent in benign nevi. By using shRNA to knockdown FHOD1 in melanoma cells, we discovered that FHOD1 depleted cells are larger, rounder and have smaller focal adhesions and inferior migratory capacity as compared to control cells. Importantly, we found FHOD1 depleted cells to have reduced colony-forming capacity and attenuated tumor growth in vivo, a finding best explained by the reduced proliferation rate caused by cell cycle arrest. Unexpectedly, FHOD1 depletion did not prevent invasive growth at the tumor margins. These results suggest that FHOD1 participates in key cellular processes that are dysregulated in malignancy, but may not be essential for melanoma cell invasion.

  18. Novel roles of formin mDia2 in lamellipodia and filopodia formation in motile cells.

    Directory of Open Access Journals (Sweden)

    Changsong Yang

    2007-11-01

    Full Text Available Actin polymerization-driven protrusion of the leading edge is a key element of cell motility. The important actin nucleators formins and the Arp2/3 complex are believed to have nonoverlapping functions in inducing actin filament bundles in filopodia and dendritic networks in lamellipodia, respectively. We tested this idea by investigating the role of mDia2 formin in leading-edge protrusion by loss-of-function and gain-of-function approaches. Unexpectedly, mDia2 depletion by short interfering RNA (siRNA severely inhibited lamellipodia. Structural analysis of the actin network in the few remaining lamellipodia suggested an mDia2 role in generation of long filaments. Consistently, constitutively active mDia2 (DeltaGBD-mDia2 induced accumulation of long actin filaments in lamellipodia and increased persistence of lamellipodial protrusion. Depletion of mDia2 also inhibited filopodia, whereas expression of DeltaGBD-mDia2 promoted their formation. Correlative light and electron microscopy showed that DeltaGBD-mDia2-induced filopodia were formed from lamellipodial network through gradual convergence of long lamellipodial filaments into bundles. Efficient filopodia induction required mDia2 targeting to the membrane, likely through a scaffolding protein Abi1. Furthermore, mDia2 and Abi1 interacted through the N-terminal regulatory sequences of mDia2 and the SH3-containing Abi1 sequences. We propose that mDia2 plays an important role in formation of lamellipodia by nucleating and/or protecting from capping lamellipodial actin filaments, which subsequently exhibit high tendency to converge into filopodia.

  19. Building bridges: formin1 of Arabidopsis forms a connection between the cell wall and the actin cytoskeleton.

    Science.gov (United States)

    Martinière, Alexandre; Gayral, Philippe; Hawes, Chris; Runions, John

    2011-04-01

    Actin microfilament (MF) organization and remodelling is critical to cell function. The formin family of actin binding proteins are involved in nucleating MFs in Arabidopsis thaliana. They all contain formin homology domains in the intracellular, C-terminal half of the protein that interacts with MFs. Formins in class I are usually targeted to the plasma membrane and this is true of Formin1 (AtFH1) of A. thaliana. In this study, we have investigated the extracellular domain of AtFH1 and we demonstrate that AtFH1 forms a bridge from the actin cytoskeleton, across the plasma membrane and is anchored within the cell wall. AtFH1 has a large, extracellular domain that is maintained by purifying selection and that contains four conserved regions, one of which is responsible for immobilising the protein. Protein anchoring within the cell wall is reduced in constructs that express truncations of the extracellular domain and in experiments in protoplasts without primary cell walls. The 18 amino acid proline-rich extracellular domain that is responsible for AtFH1 anchoring has homology with cell-wall extensins. We also have shown that anchoring of AtFH1 in the cell wall promotes actin bundling within the cell and that overexpression of AtFH1 has an inhibitory effect on organelle actin-dependant dynamics. Thus, the AtFH1 bridge provides stable anchor points for the actin cytoskeleton and is probably a crucial component of the signalling response and actin-remodelling mechanisms. © 2011 The Authors. The Plant Journal © 2011 Blackwell Publishing Ltd.

  20. Regulation of the formin Cappuccino is critical for polarity of Drosophila oocytes

    Science.gov (United States)

    Bor, Batbileg; Bois, Justin S.; Quinlan, Margot E.

    2014-01-01

    The Drosophila formin Cappuccino (Capu) creates an actin mesh-like structure that traverses the oocyte during mid-oogenesis. This mesh is thought to prevent premature onset of fast cytoplasmic streaming which normally happens during late-oogenesis. Proper cytoskeletal organization and cytoplasmic streaming are crucial for localization of polarity determinants such as osk, grk, bcd and nanos mRNAs. Capu mutants disrupt these events, leading to female sterility. Capu is regulated by another nucleator, Spire, as well as by autoinhibition in vitro. Studies in vivo confirm that Spire modulates Capu’s function in oocytes; however, how autoinhibition contributes is still unclear. To study the role of autoinhibition in flies, we expressed a Capu construct that is missing the Capu Inhibitory Domain, CapuΔN. Consistent with a gain of activity due to loss of autoinhibition, the actin mesh was denser in CapuΔN oocytes. Further, cytoplasmic streaming was delayed and fertility levels decreased. Localization of osk mRNA in early stages, and bcd and nanos in late stages, were disrupted in CapuΔN-expressing oocytes. Finally, evidence that these phenotypes were due to a loss of autoinhibition comes from co-expression of the N-terminal half of Capu with CapuΔN, which suppressed the defects in actin, cytoplasmic streaming and fertility. From these results, we conclude that Capu can be autoinhibited during Drosophila oocyte development. PMID:25557988

  1. Carcinoma associated fibroblasts (CAFs) promote breast cancer motility by suppressing mammalian Diaphanous-related formin-2 (mDia2).

    Science.gov (United States)

    Dvorak, Kaitlyn M; Pettee, Krista M; Rubinic-Minotti, Kaitlin; Su, Robin; Nestor-Kalinoski, Andrea; Eisenmann, Kathryn M

    2018-01-01

    The tumor microenvironment (TME) promotes tumor cell invasion and metastasis. An important step in the shift to a pro-cancerous microenvironment is the transformation of normal stromal fibroblasts to carcinoma-associated fibroblasts (CAFs). CAFs are present in a majority of solid tumors and can directly promote tumor cell motility via cytokine, chemokine and growth factor secretion into the TME. The exact effects that the TME has upon cytoskeletal regulation in motile tumor cells remain enigmatic. The conserved formin family of cytoskeleton regulating proteins plays an essential role in the assembly and/or bundling of unbranched actin filaments. Mammalian Diaphanous-related formin 2 (mDia2/DIAPH3/Drf3/Dia) assembles a dynamic F-actin cytoskeleton that underlies tumor cell migration and invasion. We therefore sought to understand whether CAF-derived chemokines impact breast tumor cell motility through modification of the formin-assembled F-actin cytoskeleton. In MDA-MB-231 cells, conditioned media (CM) from WS19T CAFs, a human breast tumor-adjacent CAF line, significantly and robustly increased wound closure and invasion relative to normal human mammary fibroblast (HMF)-CM. WS19T-CM also promoted proteasome-mediated mDia2 degradation in MDA-MB-231 cells relative to control HMF-CM and WS21T CAF-CM, a breast CAF cell line that failed to promote robust MDA-MB-231 migration. Cytokine array analysis of CM identified up-regulated secreted factors in WS19T relative to control WS21T CM. We identified CXCL12 as a CM factor influencing loss of mDia2 protein while increasing MDA-MB-231 cell migration. Our data suggest a mechanism whereby CAFs promote tumor cell migration and invasion through CXCL12 secretion to regulate the mDia2-directed cytoskeleton in breast tumor cells.

  2. SMIFH2-mediated mDia formin functional inhibition potentiates chemotherapeutic targeting of human ovarian cancer spheroids.

    Science.gov (United States)

    Ziske, Megan A; Pettee, Krista M; Khaing, MaNada; Rubinic, Kaitlin; Eisenmann, Kathryn M

    2016-03-25

    Due to a lack of effective screening or prevention protocol for epithelial ovarian cancer (EOC), there is a critical unmet need to develop therapeutic interventions for EOC treatment. EOC metastasis is unique. Initial dissemination is not primarily hematogenous, yet is facilitated through shedding of primary tumor cells into the peritoneal fluid and accumulating ascites. Increasingly, isolated patient spheroids point to a clinical role for spheroids in EOC metastasis. EOC spheroids are highly invasive structures that disseminate upon peritoneal mesothelium, and visceral tissues including liver and omentum. Selection for this subset of chemoresistant EOC cells could influence disease progression and/or recurrence. Thus, targeting spheroid integrity/structure may improve the chemotherapeutic responsiveness of EOC. We discovered a critical role for mammalian Diaphanous (mDia)-related formin-2 in maintaining EOC spheroid structure. Both mDia2 and the related mDia1 regulate F-actin networks critical to maintain cell-cell contacts and the integrity of multi-cellular epithelial sheets. We investigated if mDia2 functional inhibition via a small molecule inhibitor SMIFH2 combined with chemotherapeutics, such as taxol and cisplatin, inhibits the viability of EOC monolayers and clinically relevant spheroids. SMIFH2-mediated mDia formin inhibition significantly reduced both ES2 and Skov3 EOC monolayer viability while spheroid viability was minimally impacted only at the highest concentrations. Combining either cisplatin or taxol with SMIFH2 did not significantly enhance the effects of either drug alone in ES2 monolayers, while Skov3 monolayers treated with taxol or cisplatin and SMIFH2 showed significant additive inhibition of viability. ES2 spheroids were highly responsive with clear additive anti-viability effects with dual taxol or cisplatin when combined with SMIFH2 treatments. While combined taxol with SMIFH2 in spheroids showed an additive effect relative to single

  3. Interaction between cardiac myosin-binding protein C and formin Fhod3.

    Science.gov (United States)

    Matsuyama, Sho; Kage, Yohko; Fujimoto, Noriko; Ushijima, Tomoki; Tsuruda, Toshihiro; Kitamura, Kazuo; Shiose, Akira; Asada, Yujiro; Sumimoto, Hideki; Takeya, Ryu

    2018-05-08

    Mutations in cardiac myosin-binding protein C (cMyBP-C) are a major cause of familial hypertrophic cardiomyopathy. Although cMyBP-C has been considered to regulate the cardiac function via cross-bridge arrangement at the C-zone of the myosin-containing A-band, the mechanism by which cMyBP-C functions remains unclear. We identified formin Fhod3, an actin organizer essential for the formation and maintenance of cardiac sarcomeres, as a cMyBP-C-binding protein. The cardiac-specific N-terminal Ig-like domain of cMyBP-C directly interacts with the cardiac-specific N-terminal region of Fhod3. The interaction seems to direct the localization of Fhod3 to the C-zone, since a noncardiac Fhod3 variant lacking the cMyBP-C-binding region failed to localize to the C-zone. Conversely, the cardiac variant of Fhod3 failed to localize to the C-zone in the cMyBP-C-null mice, which display a phenotype of hypertrophic cardiomyopathy. The cardiomyopathic phenotype of cMyBP-C-null mice was further exacerbated by Fhod3 overexpression with a defect of sarcomere integrity, whereas that was partially ameliorated by a reduction in the Fhod3 protein levels, suggesting that Fhod3 has a deleterious effect on cardiac function under cMyBP-C-null conditions where Fhod3 is aberrantly mislocalized. Together, these findings suggest the possibility that Fhod3 contributes to the pathogenesis of cMyBP-C-related cardiomyopathy and that Fhod3 is critically involved in cMyBP-C-mediated regulation of cardiac function via direct interaction.

  4. Small-molecule intramimics of formin autoinhibition: a new strategy to target the cytoskeletal remodeling machinery in cancer cells.

    Science.gov (United States)

    Lash, L Leanne; Wallar, Bradley J; Turner, Julie D; Vroegop, Steven M; Kilkuskie, Robert E; Kitchen-Goosen, Susan M; Xu, H Eric; Alberts, Arthur S

    2013-11-15

    Although the cancer cell cytoskeleton is a clinically validated target, few new strategies have emerged for selectively targeting cell division by modulating the cytoskeletal structure, particularly ways that could avoid the cardiotoxic and neurotoxic effects of current agents such as taxanes. We address this gap by describing a novel class of small-molecule agonists of the mammalian Diaphanous (mDia)-related formins, which act downstream of Rho GTPases to assemble actin filaments, and their organization with microfilaments to establish and maintain cell polarity during migration and asymmetric division. GTP-bound Rho activates mDia family members by disrupting the interaction between the DID and DAD autoregulatory domains, which releases the FH2 domain to modulate actin and microtubule dynamics. In screening for DID-DAD disruptors that activate mDia, we identified two molecules called intramimics (IMM-01 and -02) that were sufficient to trigger actin assembly and microtubule stabilization, serum response factor-mediated gene expression, cell-cycle arrest, and apoptosis. In vivo analysis of IMM-01 and -02 established their ability to slow tumor growth in a mouse xenograft model of colon cancer. Taken together, our work establishes the use of intramimics and mDia-related formins as a new general strategy for therapeutic targeting of the cytoskeletal remodeling machinery of cancer cells. ©2013 AACR

  5. Inverted formin 2 mutations with variable expression in patients with sporadic and hereditary focal and segmental glomerulosclerosis.

    LENUS (Irish Health Repository)

    Gbadegesin, Rasheed A

    2012-01-01

    Focal and segmental glomerulosclerosis (FSGS) is a major cause of end-stage kidney disease. Recent advances in molecular genetics show that defects in the podocyte play a major role in its pathogenesis and mutations in inverted formin 2 (INF2) cause autosomal dominant FSGS. In order to delineate the role of INF2 mutations in familial and sporadic FSGS, we sought to identify variants in a large cohort of patients with FSGS. A secondary objective was to define an approach for genetic screening in families with autosomal dominant disease. A total of 248 individuals were identified with FSGS, of whom 31 had idiopathic disease. The remaining patients clustered into 64 families encompassing 15 from autosomal recessive and 49 from autosomal dominant kindreds. There were missense mutations in 8 of the 49 families with autosomal dominant disease. Three of the detected variants were novel and all mutations were confined to exon 4 of INF2, a regulatory region responsible for 90% of all changes reported in FSGS due to INF2 mutations. Thus, in our series, INF2 mutations were responsible for 16% of all cases of autosomal dominant FSGS, with these mutations clustered in exon 4. Hence, screening for these mutations may represent a rapid, non-invasive and cost-effective method for the diagnosis of autosomal dominant FSGS.

  6. Paternal effect of the nuclear formin-like protein MISFIT on Plasmodium development in the mosquito vector.

    Directory of Open Access Journals (Sweden)

    Ellen S C Bushell

    2009-08-01

    Full Text Available Malaria parasites must undergo sexual and sporogonic development in mosquitoes before they can infect their vertebrate hosts. We report the discovery and characterization of MISFIT, the first protein with paternal effect on the development of the rodent malaria parasite Plasmodium berghei in Anopheles mosquitoes. MISFIT is expressed in male gametocytes and localizes to the nuclei of male gametocytes, zygotes and ookinetes. Gene disruption results in mutant ookinetes with reduced genome content, microneme defects and altered transcriptional profiles of putative cell cycle regulators, which yet successfully invade the mosquito midgut. However, developmental arrest ensues during the ookinete transformation to oocysts leading to malaria transmission blockade. Genetic crosses between misfit mutant parasites and parasites that are either male or female gamete deficient reveal a strict requirement for a male misfit allele. MISFIT belongs to the family of formin-like proteins, which are known regulators of the dynamic remodeling of actin and microtubule networks. Our data identify the ookinete-to-oocyst transition as a critical cell cycle checkpoint in Plasmodium development and lead us to hypothesize that MISFIT may be a regulator of cell cycle progression. This study offers a new perspective for understanding the male contribution to malaria parasite development in the mosquito vector.

  7. The effect of maternal diabetes on the Wnt-PCP pathway during embryogenesis as reflected in the developing mouse eye

    Directory of Open Access Journals (Sweden)

    Beatriz López-Escobar

    2015-02-01

    Full Text Available Embryopathies that develop as a consequence of maternal diabetes have been studied intensely in both experimental and clinical scenarios. Accordingly, hyperglycaemia has been shown to downregulate the expression of elements in the non-canonical Wnt-PCP pathway, such as the Dishevelled-associated activator of morphogenesis 1 (Daam1 and Vangl2. Daam1 is a formin that is essential for actin polymerization and for cytoskeletal reorganization, and it is expressed strongly in certain organs during mouse development, including the eye, neural tube and heart. Daam1gt/gt and Daam1gt/+ embryos develop ocular defects (anophthalmia or microphthalmia that are similar to those detected as a result of hyperglycaemia. Indeed, studying the effects of maternal diabetes on the Wnt-PCP pathway demonstrated that there was strong association with the Daam1 genotype, whereby the embryopathy observed in Daam1gt/+ mutant embryos of diabetic dams was more severe. There was evidence that embryonic exposure to glucose in vitro diminishes the expression of genes in the Wnt-PCP pathway, leading to altered cytoskeletal organization, cell shape and cell polarity in the optic vesicle. Hence, the Wnt-PCP pathway appears to influence cell morphology and cell polarity, events that drive the cellular movements required for optic vesicle formation and that, in turn, are required to maintain the fate determination. Here, we demonstrate that the Wnt-PCP pathway is involved in the early stages of mouse eye development and that it is altered by diabetes, provoking the ocular phenotype observed in the affected embryos.

  8. Loss of RhoB expression enhances the myelodysplastic phenotype of mammalian diaphanous-related Formin mDia1 knockout mice.

    Directory of Open Access Journals (Sweden)

    Aaron D DeWard

    Full Text Available Myelodysplastic syndrome (MDS is characterized by ineffective hematopoiesis and hyperplastic bone marrow. Complete loss or interstitial deletions of the long arm of chromosome 5 occur frequently in MDS. One candidate tumor suppressor on 5q is the mammalian Diaphanous (mDia-related formin mDia1, encoded by DIAPH1 (5q31.3. mDia-family formins act as effectors for Rho-family small GTP-binding proteins including RhoB, which has also been shown to possess tumor suppressor activity. Mice lacking the Drf1 gene that encodes mDia1 develop age-dependent myelodysplastic features. We crossed mDia1 and RhoB knockout mice to test whether the additional loss of RhoB expression would compound the myelodysplastic phenotype. Drf1(-/-RhoB(-/- mice are fertile and develop normally. Relative to age-matched Drf1(-/-RhoB(+/- mice, the age of myelodysplasia onset was earlier in Drf1(-/-RhoB(-/- animals--including abnormally shaped erythrocytes, splenomegaly, and extramedullary hematopoiesis. In addition, we observed a statistically significant increase in the number of activated monocytes/macrophages in both the spleen and bone marrow of Drf1(-/-RhoB(-/- mice relative to Drf1(-/-RhoB(+/- mice. These data suggest a role for RhoB-regulated mDia1 in the regulation of hematopoietic progenitor cells.

  9. Drosophila homologues of adenomatous polyposis coli (APC) and the formin diaphanous collaborate by a conserved mechanism to stimulate actin filament assembly.

    Science.gov (United States)

    Jaiswal, Richa; Stepanik, Vince; Rankova, Aneliya; Molinar, Olivia; Goode, Bruce L; McCartney, Brooke M

    2013-05-10

    Vertebrate APC collaborates with Dia through its Basic domain to assemble actin filaments. Despite limited sequence homology between the vertebrate and Drosophila APC Basic domains, Drosophila APC1 collaborates with Dia to stimulate actin assembly in vitro. The mechanism of actin assembly is highly conserved over evolution. APC-Dia collaborations may be crucial in a wide range of animal cells. Adenomatous polyposis coli (APC) is a large multidomain protein that regulates the cytoskeleton. Recently, it was shown that vertebrate APC through its Basic domain directly collaborates with the formin mDia1 to stimulate actin filament assembly in the presence of nucleation barriers. However, it has been unclear whether these activities extend to homologues of APC and Dia in other organisms. Drosophila APC and Dia are each required to promote actin furrow formation in the syncytial embryo, suggesting a potential collaboration in actin assembly, but low sequence homology between the Basic domains of Drosophila and vertebrate APC has left their functional and mechanistic parallels uncertain. To address this question, we purified Drosophila APC1 and Dia and determined their individual and combined effects on actin assembly using both bulk fluorescence assays and total internal reflection fluorescence microscopy. Our data show that APC1, similar to its vertebrate homologue, bound to actin monomers and nucleated and bundled filaments. Further, Drosophila Dia nucleated actin assembly and protected growing filament barbed ends from capping protein. Drosophila APC1 and Dia directly interacted and collaborated to promote actin assembly in the combined presence of profilin and capping protein. Thus, despite limited sequence homology, Drosophila and vertebrate APCs exhibit highly related activities and mechanisms and directly collaborate with formins. These results suggest that APC-Dia interactions in actin assembly are conserved and may underlie important in vivo functions in a broad

  10. Roots of angiosperm formins: The evolutionary history of plant FH2 domain-containing proteins

    Czech Academy of Sciences Publication Activity Database

    Grunt, M.; Žárský, Viktor; Cvrčková, F.

    2008-01-01

    Roč. 8, Art_115 (2008), s. 1-19 ISSN 1471-2148 R&D Projects: GA ČR GA204/05/0268; GA MŠk(CZ) LC06004 Institutional research plan: CEZ:AV0Z50380511 Keywords : MULTIPLE SEQUENCE ALIGNMENT * TYROSINE-PHOSPHATASE * SWISS-MODEL Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 4.050, year: 2008

  11. Arabidopsis FH1 Formin Affects Cotyledon Pavement Cell Shape by Modulating Cytoskeleton Dynamics

    Czech Academy of Sciences Publication Activity Database

    Rosero, A.; Oulehlová, Denisa; Stillerová, L.; Schiebertová, P.; Gunt, M.; Žárský, Viktor; Cvrčková, F.

    2016-01-01

    Roč. 57, č. 3 (2016), s. 488-504 ISSN 0032-0781 Institutional support: RVO:61389030 Keywords : Arabidopsis thaliana * Confocal microscopy * Cotyledon pavement cells Subject RIV: ED - Physiology Impact factor: 4.760, year: 2016

  12. AtFH1 formin mutation affects actin filament and microtubule dynamics in Arabidopsis thaliana

    Czech Academy of Sciences Publication Activity Database

    Rosero, A.; Žárský, Viktor; Cvrčková, F.

    2013-01-01

    Roč. 64, č. 2 (2013), s. 585-597 ISSN 0022-0957 R&D Projects: GA ČR GAP305/10/0433 Institutional research plan: CEZ:AV0Z50380511 Keywords : Actin * Arabidopsis * At5g25500 Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 5.794, year: 2013

  13. Dinamika klyuchevykh mediatorov insulinorezistentnosti u bol'nykh sakharnym diabetom 2 tipa pri primenenii metformina (Formin Pliva?

    Directory of Open Access Journals (Sweden)

    Mikhail Ivanovich Balabolkin

    2004-12-01

    Full Text Available Цель. Изучение влияния формина (метформина не только на состояние углеводного, но и липидного обмена, динамику содержания лептина, его растворимого рецептора и альфа-фактора некроза опухолей (альфа-ФНО в сыворотке крови у больных СД 2. Материалы и методы. Обследовано 262 больных. Общий анализ крови и мочи проводили через каждый месяц наблюдения, а биохимический анализ крови, включая содержание общего ХС, ТГ, ХС ЛПНП и ЛПОНП и ХС ЛПВП, мочевины, креатинина, печеночных ферментов, проводили на 1-м и 4-м визитах. Лечение формином проводилось в течение не менее 12 нед. и изучалось при этом его влияние на состояние углеводного и липидного обмена у больных СД 2. Из общего числа обследованных у 26 больных на фоне терапии формином изучалось содержание лептина, растворимого рецептора к лептину и альфа-ФНО. Результаты. На фоне лечения формином у больных отмечалось улучшение углеводного обмена, которое сопровождалось статистически достоверным снижением гликемии натощак. Компенсация углеводного обмена у больных также сопровождалась статистически достоверным уменьшением индекса мас? сы тела с 34,02+0,33 кг/м2 до 32,73+0,33 кг/м2. У больных на фоне лечения формином наблюдается недостоверное снижение содержания лептина, растворимого рецептора к лептину и статистически достоверное снижение альфа-ФНО в сыворотке крови. Выводы. проведенные исследования показывают, что формин обладает выраженным антигипергликемическим эффектом, способствует улучшению компенсации углеводного обмена, что сопровождается статистически достоверным снижением гликемии и уровня гликозилированного гемоглобина в сыворотке крови. Наряду с этим на фоне терапии формином отмечается снижение уровня холестерина ЛПНП и повышение содержания холестерина ЛПВП, при практически неизмененных показателях содержания общего холестерина в сыворотке крови. Кроме того, лечение формином способствует статистически достоверному снижению содержания альфа-ФНО и умеренному понижению уровня лептина и его растворимого рецептора в сыворотке крови.

  14. Unique and Overlapping Functions of Formins Frl and DAAM During Ommatidial Rotation and Neuronal Development in Drosophila

    OpenAIRE

    Dollar, Gretchen; Gombos, Rita; Barnett, Austen A.; Sanchez Hernandez, David; Maung, Saw M. T.; Mih?ly, Jozsef; Jenny, Andreas

    2016-01-01

    The noncanonical Frizzled/planar cell polarity (PCP) pathway regulates establishment of polarity within the plane of an epithelium to generate diversity of cell fates, asymmetric, but highly aligned structures, or to orchestrate the directional migration of cells during convergent extension during vertebrate gastrulation. In Drosophila, PCP signaling is essential to orient actin wing hairs and to align ommatidia in the eye, in part by coordinating the movement of groups of photoreceptor cells...

  15. Expression of GFP-mTalin reveals an actin-related role for the Arabidopsis Class II formin AtFH12

    Czech Academy of Sciences Publication Activity Database

    Cvrčková, F.; Grunt, M.; Žárský, Viktor

    2012-01-01

    Roč. 56, č. 3 (2012), s. 431-440 ISSN 0006-3134 R&D Projects: GA ČR GAP305/10/0433 Institutional research plan: CEZ:AV0Z50380511 Keywords : FH2 proteins * genetic redundancy * salt stress Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 1.692, year: 2012

  16. The plant formin AtFH4 interacts with both actin and microtubules, and contains a newly identified microtubule-binding domain

    Czech Academy of Sciences Publication Activity Database

    Deeks, M.J.; Fendrych, Matyáš; Smertenko, A.; Bell, K.S.; Oparka, K.; Cvrčková, F.; Žárský, Viktor; Hussey, P.J.

    2010-01-01

    Roč. 123, č. 8 (2010), s. 1209-1215 ISSN 0021-9533 R&D Projects: GA MŠk(CZ) LC06004; GA ČR GAP305/10/0433 Institutional research plan: CEZ:AV0Z50380511 Keywords : Actin regulating proteins * Membrane * Microtubule Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 6.290, year: 2010

  17. Desulfotomaculum arcticum sp nov., a novel spore-formin, moderately thermophilic, sulfate-reducing bacterium isolated from a permanently cold fjord sediment of Svalbard

    DEFF Research Database (Denmark)

    Vandieken, V.; Knoblauch, C.; Jørgensen, BB

    2006-01-01

    Strain 15 T is a novel spore-forming, sulfate-reducing bacterium isolated from a permanently cold fjord sediment of Svalbard. Sulfate could be replaced by sulfite or thiosulfate. Hydrogen, formate, lactate, propionate, butyrate, hexanoate, methanol, ethanol, propanol, butanol, pyruvate, malate, s...... related to Desulfotomaculum thermosapovorans MLF(T) (93-5% 16S rRNA gene sequence similarity). Strain 15 T represents a novel species, for which the name Desulfotomaculurn arcticum sp. nov. is proposed. The type strain is strain 15 T (=DSM 17038(T)=jCM 12923(T))....

  18. Arabidopsis group Ie formins localize to specific cell membrane domains, interact with actin-binding proteins and cause defects in cell expansion upon aberrant expression

    Czech Academy of Sciences Publication Activity Database

    Deeks, M.J.; Cvrčková, F.; Machesky, M. L.; Mikitova, V.; Ketelaar, T.; Žárský, Viktor; Davies, B.; Hussey, P.J.

    2005-01-01

    Roč. 168, č. 3 (2005), s. 529-540 ISSN 0028-646X R&D Projects: GA ČR GA204/02/1461; GA ČR GA204/05/0268 Institutional research plan: CEZ:AV0Z50380511 Keywords : actin * Arabidopsis * cytoskeleton Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 4.285, year: 2005

  19. Technical Note: Impact of the geometry dependence of the ion chamber detector response function on a convolution-based method to address the volume averaging effect

    Energy Technology Data Exchange (ETDEWEB)

    Barraclough, Brendan; Lebron, Sharon [Department of Radiation Oncology, University of Florida, Gainesville, Florida 32608 and J. Crayton Pruitt Family Department of Biomedical Engineering, University of Florida, Gainesville, Florida 32611 (United States); Li, Jonathan G.; Fan, Qiyong; Liu, Chihray; Yan, Guanghua, E-mail: yangua@shands.ufl.edu [Department of Radiation Oncology, University of Florida, Gainesville, Florida 32608 (United States)

    2016-05-15

    Purpose: To investigate the geometry dependence of the detector response function (DRF) of three commonly used scanning ionization chambers and its impact on a convolution-based method to address the volume averaging effect (VAE). Methods: A convolution-based approach has been proposed recently to address the ionization chamber VAE. It simulates the VAE in the treatment planning system (TPS) by iteratively convolving the calculated beam profiles with the DRF while optimizing the beam model. Since the convolved and the measured profiles are subject to the same VAE, the calculated profiles match the implicit “real” ones when the optimization converges. Three DRFs (Gaussian, Lorentzian, and parabolic function) were used for three ionization chambers (CC04, CC13, and SNC125c) in this study. Geometry dependent/independent DRFs were obtained by minimizing the difference between the ionization chamber-measured profiles and the diode-measured profiles convolved with the DRFs. These DRFs were used to obtain eighteen beam models for a commercial TPS. Accuracy of the beam models were evaluated by assessing the 20%–80% penumbra width difference (PWD) between the computed and diode-measured beam profiles. Results: The convolution-based approach was found to be effective for all three ionization chambers with significant improvement for all beam models. Up to 17% geometry dependence of the three DRFs was observed for the studied ionization chambers. With geometry dependent DRFs, the PWD was within 0.80 mm for the parabolic function and CC04 combination and within 0.50 mm for other combinations; with geometry independent DRFs, the PWD was within 1.00 mm for all cases. When using the Gaussian function as the DRF, accounting for geometry dependence led to marginal improvement (PWD < 0.20 mm) for CC04; the improvement ranged from 0.38 to 0.65 mm for CC13; for SNC125c, the improvement was slightly above 0.50 mm. Conclusions: Although all three DRFs were found adequate to

  20. Design and validation of an open-source library of dynamic reference frames for research and education in optical tracking.

    Science.gov (United States)

    Brown, Alisa; Uneri, Ali; Silva, Tharindu De; Manbachi, Amir; Siewerdsen, Jeffrey H

    2018-04-01

    Dynamic reference frames (DRFs) are a common component of modern surgical tracking systems; however, the limited number of commercially available DRFs poses a constraint in developing systems, especially for research and education. This work presents the design and validation of a large, open-source library of DRFs compatible with passive, single-face tracking systems, such as Polaris stereoscopic infrared trackers (NDI, Waterloo, Ontario). An algorithm was developed to create new DRF designs consistent with intra- and intertool design constraints and convert to computer-aided design (CAD) files suitable for three-dimensional printing. A library of 10 such groups, each with 6 to 10 DRFs, was produced and tracking performance was validated in comparison to a standard commercially available reference, including pivot calibration, fiducial registration error (FRE), and target registration error (TRE). Pivot tests showed calibration error [Formula: see text], indistinguishable from the reference. FRE was [Formula: see text], and TRE in a CT head phantom was [Formula: see text], both equivalent to the reference. The library of DRFs offers a useful resource for surgical navigation research and could be extended to other tracking systems and alternative design constraints.

  1. Tumor gene expression and prognosis in breast cancer patients with 10 or more positive lymph nodes.

    Science.gov (United States)

    Cobleigh, Melody A; Tabesh, Bita; Bitterman, Pincas; Baker, Joffre; Cronin, Maureen; Liu, Mei-Lan; Borchik, Russell; Mosquera, Juan-Miguel; Walker, Michael G; Shak, Steven

    2005-12-15

    This study, along with two others, was done to develop the 21-gene Recurrence Score assay (Oncotype DX) that was validated in a subsequent independent study and is used to aid decision making about chemotherapy in estrogen receptor (ER)-positive, node-negative breast cancer patients. Patients with >or=10 nodes diagnosed from 1979 to 1999 were identified. RNA was extracted from paraffin blocks, and expression of 203 candidate genes was quantified using reverse transcription-PCR (RT-PCR). Seventy-eight patients were studied. As of August 2002, 77% of patients had distant recurrence or breast cancer death. Univariate Cox analysis of clinical and immunohistochemistry variables indicated that HER2/immunohistochemistry, number of involved nodes, progesterone receptor (PR)/immunohistochemistry (% cells), and ER/immunohistochemistry (% cells) were significantly associated with distant recurrence-free survival (DRFS). Univariate Cox analysis identified 22 genes associated with DRFS. Higher expression correlated with shorter DRFS for the HER2 adaptor GRB7 and the macrophage marker CD68. Higher expression correlated with longer DRFS for tumor protein p53-binding protein 2 (TP53BP2) and the ER axis genes PR and Bcl2. Multivariate methods, including stepwise variable selection and bootstrap resampling of the Cox proportional hazards regression model, identified several genes, including TP53BP2 and Bcl2, as significant predictors of DRFS. Tumor gene expression profiles of archival tissues, some more than 20 years old, provide significant information about risk of distant recurrence even among patients with 10 or more nodes.

  2. Implementation of the New Approach for the Dose-Response Functions Development for the Case of Athens and Greece

    Science.gov (United States)

    Christodoulakis, J.; Tzanis, C. G.; Varotsos, C. A.; Kouremadas, G.

    2016-08-01

    Dose-response functions (DRFs) are functions used for estimating corrosion and/or soiling levels of materials used in constructions and cultural monuments. In order to achieve this, DRFs lean on ground-based measurements of specific air pollution and climatic parameters like nitrogen oxides, ozone, temperature and others. In DRAGON 3 2015 Symposium we presented a new approach which proposed a technique for using satellite-based data for the necessary parameters instead of ground-based expanding in this way: a) the usage of DRFs in cases/areas where there is no availability of in situ measurements, b) the applicability of satellite-based data. In this work we present mapping results of deterioration levels (corrosion and soiling) for the case of Athens, Greece but also for the whole Greece country.

  3. DRF3 as a Cholesterol-Dependent Regulator of Src in Prostate Cancer

    National Research Council Canada - National Science Library

    Freeman, Michael R

    2007-01-01

    This project focuses on the novel finding from our group that the formin protein, Drf3, is a signaling molecule positioned downstream from the EGF receptor that intersects with the tyrosine kinase Src...

  4. Deep radiomic prediction with clinical predictors of the survival in patients with rheumatoid arthritis-associated interstitial lung diseases

    Science.gov (United States)

    Nasirudina, Radin A.; Näppi, Janne J.; Watari, Chinatsu; Matsuhiro, Mikio; Hironaka, Toru; Kido, Shoji; Yoshida, Hiroyuki

    2018-02-01

    We developed and evaluated the effect of our deep-learning-derived radiomic features, called deep radiomic features (DRFs), together with their combination with clinical predictors, on the prediction of the overall survival of patients with rheumatoid arthritis-associated interstitial lung disease (RA-ILD). We retrospectively identified 70 RA-ILD patients with thin-section lung CT and pulmonary function tests. An experienced observer delineated regions of interest (ROIs) from the lung regions on the CT images, and labeled them into one of four ILD patterns (ground-class opacity, reticulation, consolidation, and honeycombing) or a normal pattern. Small image patches centered at individual pixels on these ROIs were extracted and labeled with the class of the ROI to which the patch belonged. A deep convolutional neural network (DCNN), which consists of a series of convolutional layers for feature extraction and a series of fully connected layers, was trained and validated with 5-fold cross-validation for classifying the image patches into one of the above five patterns. A DRF vector for each patch was identified as the output of the last convolutional layer of the DCNN. Statistical moments of each element of the DRF vectors were computed to derive a DRF vector that characterizes the patient. The DRF vector was subjected to a Cox proportional hazards model with elastic-net penalty for predicting the survival of the patient. Evaluation was performed by use of bootstrapping with 2,000 replications, where concordance index (C-index) was used as a comparative performance metric. Preliminary results on clinical predictors, DRFs, and their combinations thereof showed (a) Gender and Age: C-index 64.8% [95% confidence interval (CI): 51.7, 77.9]; (b) gender, age, and physiology (GAP index): C-index: 78.5% [CI: 70.50 86.51], P < 0.0001 in comparison with (a); (c) DRFs: C-index 85.5% [CI: 73.4, 99.6], P < 0.0001 in comparison with (b); and (d) DRF and GAP: C-index 91.0% [CI: 84

  5. Air Pathway Dose Modeling for the E-Area Low-Level Waste Facility

    Energy Technology Data Exchange (ETDEWEB)

    Dixon, K. L. [Savannah River Site (SRS), Aiken, SC (United States); Minter, K. M. [Savannah River Site (SRS), Aiken, SC (United States)

    2017-09-06

    Dose-release factors (DRFs) were calculated for potential atmospheric releases of several radionuclides from the E-Area Low-Level Waste Facility (ELLWF). The ELLWF receives solid low-level radioactive waste from across the Savannah River Site (SRS) and offsite for disposal. These factors represent the maximum dose a receptor would receive if standing at either 100 m or 11,410 m (Site Boundary) from the edge of an ELLWF disposal unit which are points of assessment (POA) for Department of Energy (DOE) Order 435.1 performance assessments (PA). The DRFs were calculated for 1 Ci of the specified radionuclide being released from the ground surface to the atmosphere (mrem per curie released). The calculation conservatively represented the ELLWF as a point source, and conservatively assumed the receptor was positioned at the center of the contaminant plume and continuously exposed for a period of one year. These DRFs can be refined to take into consideration disposal unit size, proximity and timing of peak dose to establish less conservative radionuclide specific disposal limits. DRFs were calculated for H-3 and C-14 in Revision 0 of this report. H-3 as HTO and C-14 as CO2 were identified as volatile radionuclides of potential concern in earlier radionuclide screening studies. In Revision 1, DRFs were calculated for eight additional radionuclides identified by an updated screening analysis as potentially important volatile radionuclides. These include Ar-37, Ar-39, Ar-42, Hg-194, Hg- 203, Kr-81, Kr-85, and Xe-127.

  6. NESHAP Area-Specific Dose-Release Factors for Potential Onsite Member-of-the-Public Locations at SRS using CAP88-PC Version 4.0

    Energy Technology Data Exchange (ETDEWEB)

    Trimor, P. [Savannah River Site (SRS), Aiken, SC (United States). Savannah River National Lab. (SRNL)

    2017-08-09

    The Environmental Protection Agency (EPA) requires the use of the computer model CAP88-PC to estimate the total effective doses (TED) for demonstrating compliance with 40 CFR 61, Subpart H (EPA 2006), the National Emission Standards for Hazardous Air Pollutants (NESHAP) regulations. As such, CAP88 Version 4.0 was used to calculate the receptor dose due to routine atmospheric releases at the Savannah River Site (SRS). For estimation, NESHAP dose-release factors (DRFs) have been supplied to Environmental Compliance and Area Closure Projects (EC&ACP) for many years. DRFs represent the dose to a maximum receptor exposed to 1 Ci of a specified radionuclide being released into the atmosphere. They are periodically updated to include changes in the CAP88 version, input parameter values, site meteorology, and location of the maximally exposed individual (MEI). In this report, the DRFs were calculated for potential radionuclide atmospheric releases from 13 SRS release points. The three potential onsite MEI locations to be evaluated are B-Area, Three Rivers Landfill (TRL), and Savannah River Ecology Lab Conference Center (SRELCC) with TRL’s onsite workers considered as members-of-the-public, and the potential future constructions of dormitories at SRELCC and Barracks at B-Area. Each MEI location was evaluated at a specified compass sector with different area to receptor distances and was conducted for both ground-level and elevated release points. The analysis makes use of area-specific meteorological data (Viner 2014). The resulting DRFs are compared to the 2014 NESHAP offsite MEI DRFs for three operational areas; A-Area, H-Area, and COS for a release rate of 1 Ci of tritium oxide at 0 ft. elevation. CAP88 was executed again using the 2016 NESHAP MEI release rates for 0 and 61 m stack heights to determine the radionuclide dose at TRL from the center-of-site (COS).

  7. Footprints of air pollution and changing environment on the sustainability of built infrastructure.

    Science.gov (United States)

    Kumar, Prashant; Imam, Boulent

    2013-02-01

    Over 150 research articles relating three multi-disciplinary topics (air pollution, climate change and civil engineering structures) are reviewed to examine the footprints of air pollution and changing environment on the sustainability of building and transport structures (referred as built infrastructure). The aim of this review is to synthesize the existing knowledge on this topic, highlight recent advances in our understanding and discuss research priorities. The article begins with the background information on sources and emission trends of global warming (CO(2), CH(4), N(2)O, CFCs, SF(6)) and corrosive (SO(2), O(3), NO(X)) gases and their role in deterioration of building materials (e.g. steel, stone, concrete, brick and wood) exposed in outdoor environments. Further section covers the impacts of climate- and pollution-derived chemical pathways, generally represented by dose-response functions (DRFs), and changing environmental conditions on built infrastructure. The article concludes with the discussions on the topic areas covered and research challenges. A comprehensive inventory of DRFs is compiled. The case study carried out for analysing the inter-comparability of various DRFs on four different materials (carbon steel, limestone, zinc and copper) produced comparable results. Results of another case study revealed that future projected changes in temperature and/or relatively humidity are expected to have a modest effect on the material deterioration rate whereas changes in precipitation were found to show a more dominant impact. Evidences suggest that both changing and extreme environmental conditions are expected to affect the integrity of built infrastructure both in terms of direct structural damage and indirect losses of transport network functionality. Unlike stone and metals, substantially limited information is available on the deterioration of brick, concrete and wooden structures. Further research is warranted to develop more robust and

  8. Development of a simple detector response function generation program: The CEARDRFs code

    Energy Technology Data Exchange (ETDEWEB)

    Wang Jiaxin, E-mail: jwang3@ncsu.edu [Center for Engineering Applications of Radioisotopes (CEAR), Department of Nuclear Engineering, North Carolina State University, Raleigh, NC 27695 (United States); Wang Zhijian; Peeples, Johanna [Center for Engineering Applications of Radioisotopes (CEAR), Department of Nuclear Engineering, North Carolina State University, Raleigh, NC 27695 (United States); Yu Huawei [Center for Engineering Applications of Radioisotopes (CEAR), Department of Nuclear Engineering, North Carolina State University, Raleigh, NC 27695 (United States); College of Geo-Resources and Information, China University of Petroleum, Qingdao, Shandong 266555 (China); Gardner, Robin P. [Center for Engineering Applications of Radioisotopes (CEAR), Department of Nuclear Engineering, North Carolina State University, Raleigh, NC 27695 (United States)

    2012-07-15

    A simple Monte Carlo program named CEARDRFs has been developed to generate very accurate detector response functions (DRFs) for scintillation detectors. It utilizes relatively rigorous gamma-ray transport with simple electron transport, and accounts for two phenomena that have rarely been treated: scintillator non-linearity and the variable flat continuum part of the DRF. It has been proven that these physics and treatments work well for 3 Multiplication-Sign 3 Double-Prime and 6 Multiplication-Sign 6 Double-Prime cylindrical NaI detector in CEAR's previous work. Now this approach has been expanded to cover more scintillation detectors with various common shapes and sizes. Benchmark experiments of 2 Multiplication-Sign 2 Double-Prime cylindrical BGO detector and 2 Multiplication-Sign 4 Multiplication-Sign 16 Double-Prime rectangular NaI detector have been carried out at CEAR with various radiactive sources. The simulation results of CEARDRFs have also been compared with MCNP5 calculations. The benchmark and comparison show that CEARDRFs can generate very accurate DRFs (more accurate than MCNP5) at a very fast speed (hundred times faster than MCNP5). The use of this program can significantly increase the accuracy of applications relying on detector spectroscopy like prompt gamma-ray neutron activation analysis, X-ray fluorescence analysis, oil well logging and homeland security. - Highlights: Black-Right-Pointing-Pointer CEARDRF has been developed to generate detector response functions (DRFs) for scintillation detectors a. Black-Right-Pointing-Pointer Generated DRFs are very accurate. Black-Right-Pointing-Pointer Simulation speed is hundreds of times faster than MCNP5. Black-Right-Pointing-Pointer It utilizes rigorous gamma-ray transport with simple electron transport. Black-Right-Pointing-Pointer It also accounts for scintillator non-linearity and the variable flat continuum part.

  9. Development of a simple detector response function generation program: The CEARDRFs code

    International Nuclear Information System (INIS)

    Wang Jiaxin; Wang Zhijian; Peeples, Johanna; Yu Huawei; Gardner, Robin P.

    2012-01-01

    A simple Monte Carlo program named CEARDRFs has been developed to generate very accurate detector response functions (DRFs) for scintillation detectors. It utilizes relatively rigorous gamma-ray transport with simple electron transport, and accounts for two phenomena that have rarely been treated: scintillator non-linearity and the variable flat continuum part of the DRF. It has been proven that these physics and treatments work well for 3×3″ and 6×6″ cylindrical NaI detector in CEAR's previous work. Now this approach has been expanded to cover more scintillation detectors with various common shapes and sizes. Benchmark experiments of 2×2″ cylindrical BGO detector and 2×4×16″ rectangular NaI detector have been carried out at CEAR with various radiactive sources. The simulation results of CEARDRFs have also been compared with MCNP5 calculations. The benchmark and comparison show that CEARDRFs can generate very accurate DRFs (more accurate than MCNP5) at a very fast speed (hundred times faster than MCNP5). The use of this program can significantly increase the accuracy of applications relying on detector spectroscopy like prompt gamma-ray neutron activation analysis, X-ray fluorescence analysis, oil well logging and homeland security. - Highlights: ► CEARDRF has been developed to generate detector response functions (DRFs) for scintillation detectors a. ► Generated DRFs are very accurate. ► Simulation speed is hundreds of times faster than MCNP5. ► It utilizes rigorous gamma-ray transport with simple electron transport. ► It also accounts for scintillator non-linearity and the variable flat continuum part.

  10. Is Mastectomy Superior to Breast-Conserving Treatment for Young Women?

    International Nuclear Information System (INIS)

    Coulombe, Genevieve; Tyldesley, Scott; Speers, Caroline B.A.; Paltiel, Chuck M.Sc.; Aquino-Parsons, Christina; Bernstein, Vanessa; Truong, Pauline T.; Keyes, Mira; Olivotto, Ivo A.

    2007-01-01

    Purpose: To examine whether modified radical mastectomy (MRM) improves outcomes compared with breast-conserving treatment (BCT) in young women. Methods and Materials: Women aged 20-49 years, diagnosed with early breast cancer between 1989 and 1998, were identified. Management with BCT or MRM was compared for local (L), locoregional (LR), and distant relapse-free survival (DRFS) and breast cancer-specific survival (BCSS) by age group (20-39 years, 40-49 years). The analysis was repeated for patients considered 'ideal' candidates for BCT: tumor size ≤2 cm, pathologically negative axillary nodes, negative margins, and no reported ductal carcinoma in situ. Results: A total of 1,597 women received BCT, and 801 had MRM. After a median follow-up of 9.0 years, the outcomes (L, LR, BCSS) were worse for the younger age group; however, the outcomes were not statistically different by type of local treatment. For women aged 20-39 years considered 'ideal' for BCT, those treated with BCT had slightly lower LRFS compared with those treated with MRM (p = 0.3), but DRFS and BCSS were similar. Conclusions: A difference in LRFS at 10 years potentially favored MRM among women aged 20-39 years considered 'ideal' BCT candidates but was not statistically significant and did not translate into a noticeable difference in DRFS or BCSS. Our data suggest that young age alone is not a contraindication to BCT

  11. NESHAP Dose-Release Factor Isopleths for Five Source-to-Receptor Distances from the Center of Site and H-Area for all Compass Sectors at SRS using CAP88-PC Version 4.0

    Energy Technology Data Exchange (ETDEWEB)

    Trimor, P. [Savannah River Site (SRS), Aiken, SC (United States). Savannah River National Lab. (SRNL)

    2017-08-09

    The Environmental Protection Agency (EPA) requires the use of the computer model CAP88-PC to estimate the total effective doses (TED) for demonstrating compliance with 40 CFR 61, Subpart H (EPA 2006), the National Emission Standards for Hazardous Air Pollutants (NESHAP) regulations. As such, CAP88 Version 4.0 was used to calculate the receptor dose due to routine atmospheric releases at the Savannah River Site (SRS). For estimation, NESHAP dose-release factors (DRFs) have been supplied to Environmental Compliance and Area Closure Projects (EC&ACP) for many years. DRFs represent the dose to a maximum receptor exposed to 1 Ci of a specified radionuclide being released into the atmosphere. They are periodically updated to include changes in the CAP88 version, input parameter values, site meteorology, and location of the maximally exposed individual (MEI). This report presents the DRFs of tritium oxide released at two onsite locations, center-of-site (COS) and H-Area, at 0 ft. elevation to maximally exposed individuals (MEIs) located 1000, 3000, 6000, 9000, and 12000 meters from the release areas for 16 compass sectors. The analysis makes use of area-specific meteorological data (Viner 2014).

  12. GENOMIC PREDICTOR OF RESPONSE AND SURVIVAL FOLLOWING TAXANE-ANTHRACYCLINE CHEMOTHERAPY FOR INVASIVE BREAST CANCER

    Science.gov (United States)

    Hatzis, Christos; Pusztai, Lajos; Valero, Vicente; Booser, Daniel J.; Esserman, Laura; Lluch, Ana; Vidaurre, Tatiana; Holmes, Frankie; Souchon, Eduardo; Martin, Miguel; Cotrina, José; Gomez, Henry; Hubbard, Rebekah; Chacón, J. Ignacio; Ferrer-Lozano, Jaime; Dyer, Richard; Buxton, Meredith; Gong, Yun; Wu, Yun; Ibrahim, Nuhad; Andreopoulou, Eleni; Ueno, Naoto T.; Hunt, Kelly; Yang, Wei; Nazario, Arlene; DeMichele, Angela; O’Shaughnessy, Joyce; Hortobagyi, Gabriel N.; Symmans, W. Fraser

    2017-01-01

    CONTEXT Accurate prediction of who will (or won’t) have high probability of survival benefit from standard treatments is fundamental for individualized cancer treatment strategies. OBJECTIVE To develop a predictor of response and survival from chemotherapy for newly diagnosed invasive breast cancer. DESIGN Development of different predictive signatures for resistance and response to neoadjuvant chemotherapy (stratified according to estrogen receptor (ER) status) from gene expression microarrays of newly diagnosed breast cancer (310 patients). Then prediction of breast cancer treatment-sensitivity using the combination of signatures for: 1) sensitivity to endocrine therapy, 2) chemo-resistance, and 3) chemo-sensitivity. Independent validation (198 patients) and comparison with other reported genomic predictors of chemotherapy response. SETTING Prospective multicenter study to develop and test genomic predictors for neoadjuvant chemotherapy. PATIENTS Newly diagnosed HER2-negative breast cancer treated with chemotherapy containing sequential taxane and anthracycline-based regimens then endocrine therapy (if hormone receptor-positive). MAIN OUTCOME MEASURES Distant relapse-free survival (DRFS) if predicted treatment-sensitive and absolute risk reduction (ARR, difference in DRFS of the two predicted groups) at median follow-up (3 years), and their 95% confidence intervals (CI). RESULTS Patients in the independent validation cohort (99% clinical Stage II–III) who were predicted to be treatment-sensitive (28% of total) had DRFS of 92% (CI 85–100) and survival benefit compared to others (absolute risk reduction (ARR) 18%; CI 6–28). Predictions were accurate if breast cancer was ER-positive (30% predicted sensitive, DRFS 97%, CI 91–100; ARR 11%, CI 0.1–21) or ER-negative (26% predicted sensitive, DRFS 83%, CI 68–100; ARR 26%, CI 4–28), and were significant in multivariate analysis after adjusting for relevant clinical-pathologic characteristics. Other

  13. Dgroup: DG01684 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available DG01684 DGroup Biguanide antidiabetic -formin DG01684.gif DG00112 ... Phenformin ... D08... hydrochloride (JP17) ... D04103 ... Etoformin hydrochloride (USAN) Antidiabetic agen...t ... DG01685 ... Insulin sensitizer Unclassified ... DG02044 ... Hypoglycemics ATC code: A10BA Antidiabetics AMPK (PRKAA) [HSA:5562 5563] [KO:K07198] ...

  14. Profilin-Dependent Nucleation and Assembly of Actin Filaments Controls Cell Elongation in Arabidopsis1[OPEN

    Science.gov (United States)

    Cao, Lingyan; Blanchoin, Laurent; Staiger, Christopher J.

    2016-01-01

    Actin filaments in plant cells are incredibly dynamic; they undergo incessant remodeling and assembly or disassembly within seconds. These dynamic events are choreographed by a plethora of actin-binding proteins, but the exact mechanisms are poorly understood. Here, we dissect the contribution of Arabidopsis (Arabidopsis thaliana) PROFILIN1 (PRF1), a conserved actin monomer-binding protein, to actin organization and single filament dynamics during axial cell expansion of living epidermal cells. We found that reduced PRF1 levels enhanced cell and organ growth. Surprisingly, we observed that the overall frequency of nucleation events in prf1 mutants was dramatically decreased and that a subpopulation of actin filaments that assemble at high rates was reduced. To test whether profilin cooperates with plant formin proteins to execute actin nucleation and rapid filament elongation in cells, we used a pharmacological approach. Here, we used Small Molecule Inhibitor of Formin FH2 (SMIFH2), after validating its mode of action on a plant formin in vitro, and observed a reduced nucleation frequency of actin filaments in live cells. Treatment of wild-type epidermal cells with SMIFH2 mimicked the phenotype of prf1 mutants, and the nucleation frequency in prf1-2 mutant was completely insensitive to these treatments. Our data provide compelling evidence that PRF1 coordinates the stochastic dynamic properties of actin filaments by modulating formin-mediated actin nucleation and assembly during plant cell expansion. PMID:26574597

  15. AcEST: DK952739 [AcEST

    Lifescience Database Archive (English)

    Full Text Available BSNV Structural polyprotein OS=Blotched snakehead... 31 4.0 sp|Q7XWS7|FH12_ORYSJ Formin-like protein 12 OS=O...GGGTTGETSKSPPDEPDDSEAGSVSSMPPLEG 2401 >sp|Q8AZM0|POLS_BSNV Structural polyprotein OS=Blotched snakehead viru

  16. AcEST: DK963597 [AcEST

    Lifescience Database Archive (English)

    Full Text Available ral polyprotein OS=Blotched snakehead... 31 5.0 sp|Q7XWS7|FH12_ORYSJ Formin-like protein 12 OS=Oryza sativa ...GSVSSMPPLEG 2401 >sp|Q8AZM0|POLS_BSNV Structural polyprotein OS=Blotched snakehead virus PE=1 SV=1 Length =

  17. AcEST: DK962818 [AcEST

    Lifescience Database Archive (English)

    Full Text Available _BSNV Structural polyprotein OS=Blotched snakehead... 31 5.2 sp|Q7XWS7|FH12_ORYSJ Formin-like protein 12 OS=...SKSPPDEPDDSEAGSVSSMPPLEG 2401 >sp|Q8AZM0|POLS_BSNV Structural polyprotein OS=Blotched snakehead virus PE=1 S

  18. AcEST: DK952719 [AcEST

    Lifescience Database Archive (English)

    Full Text Available sp|Q8AZM0|POLS_BSNV Structural polyprotein OS=Blotched snakehead... 31 4.3 sp|Q7XWS7|FH12_ORYSJ Formin-like...---------SGDPGHSTGGGTTGETSKSPPDEPDDSEAGSVSSMPPLEG 2401 >sp|Q8AZM0|POLS_BSNV Structural polyprotein OS=Blotched snake

  19. AcEST: DK954087 [AcEST

    Lifescience Database Archive (English)

    Full Text Available 2 2.1 sp|Q8AZM0|POLS_BSNV Structural polyprotein OS=Blotched snakehead... 31 3.6 sp|Q7XWS7|FH12_ORYSJ Formin...uery: 326 KRLDGAAEAANAGY 367 + EA NA Y Sbjct: 322 DEIVKLVEARNAKY 335 >sp|Q8AZM0|POLS_BSNV Structural polyprotein OS=Blotched snake

  20. AcEST: DK949303 [AcEST

    Lifescience Database Archive (English)

    Full Text Available POLS_BSNV Structural polyprotein OS=Blotched snakehead... 31 5.6 sp|Q7XWS7|FH12_ORYSJ Formin-like protein 12...TGETSKSPPDEPDDSEAGSVSSMPPLEG 2401 >sp|Q8AZM0|POLS_BSNV Structural polyprotein OS=Blotched snakehead virus PE

  1. FMNL2 and -3 regulate Golgi architecture and anterograde transport downstream of Cdc42

    DEFF Research Database (Denmark)

    Kage, Frieda; Steffen, Anika; Ellinger, Adolf

    2017-01-01

    The Rho-family small GTPase Cdc42 localizes at plasma membrane and Golgi complex and aside from protrusion and migration operates in vesicle trafficking, endo- and exocytosis as well as establishment and/or maintenance of cell polarity. The formin family members FMNL2 and -3 are actin assembly fa...

  2. DRF3 as a Cholesterol Dependent Regulator of Src in Prostate Cancer

    Science.gov (United States)

    2009-01-01

    blebbing ( Hannemann et al., 2008). Human DRF3 is not well studied, although analyses of the mouse homolog Drf3, and the close mouse paralog, Drf1/mDia1... Hannemann S, Madrid R, Stastna J, et al. The diaphanous related formin FHOD1 associates with ROCK1 and promotes Src-dependent plasma membrane blebbing

  3. Methods for Isolation, Purification, and Propagation of Bacteriophages of Campylobacter jejuni

    DEFF Research Database (Denmark)

    Gencay, Yilmaz Emre; Birk, Tina; Sørensen, Martine Camilla Holst

    2017-01-01

    Here, we describe the methods for isolation, purification, and propagation of Campylobacter jejuni bacteriophages from samples expected to contain high number of phages such as chicken feces. The overall steps are (1) liberation of phages from the sample material; (2) observation of plaque-formin...

  4. Assessment of "YouTube" Content for Distal Radius Fracture Immobilization.

    Science.gov (United States)

    Addar, Abdullah; Marwan, Yousef; Algarni, Nizar; Berry, Gregory

    Distal radius fractures (DRFs) are the most common orthopedic fractures, with >70% of cases treated by closed immobilization using a short arm cast or a sugar tong splint. However, inadequate immobilization is a risk factor for loss of reduction requiring repeat reduction or surgical treatment. Therefore, education of clinical skills for appropriate immobilization of DRFs is important. With the increasing use of web-based information by medical learners, our aim was to assess the quality and quantity of videos regarding closed immobilization of DRFs on YouTube. Retrospective review of YouTube videos on distal radius fracture immobilization using specific search terms. Identified videos were analyzed for their educational value, quality of the technical skill demonstrated, and overall metrics. Educational value was scored on a 5-point scale, with "1" indicative of low quality and "5" of high quality. Not applicable. Among the 68,366 videos identified, 16 met our inclusion criteria of being in English; performed by a health care professional or institution; and with casting being the major theme of the educational information provided. Of these 16 videos, 6 had an educational value score of 4 or 5, with the remaining 10 having a score ≤3. Although immobilization was demonstrated by cast technician specialized in orthopedics, skills were also performed by orthopedic attendants, urgent care physicians, orthopedic residents, and nurse practitioners. The credentials of the performer in 3 videos were not identified. There is a need to promote high-quality educational videos produced by established medical school faculty members on open, web-based, portals. Copyright © 2017 Association of Program Directors in Surgery. Published by Elsevier Inc. All rights reserved.

  5. Improvement of single detector proton radiography by incorporating intensity of time-resolved dose rate functions

    Science.gov (United States)

    Zhang, Rongxiao; Jee, Kyung-Wook; Cascio, Ethan; Sharp, Gregory C.; Flanz, Jacob B.; Lu, Hsiao-Ming

    2018-01-01

    Proton radiography, which images patients with the same type of particles as those with which they are to be treated, is a promising approach to image guidance and water equivalent path length (WEPL) verification in proton radiation therapy. We have shown recently that proton radiographs could be obtained by measuring time-resolved dose rate functions (DRFs) using an x-ray amorphous silicon flat panel. The WEPL values were derived solely from the root-mean-square (RMS) of DRFs, while the intensity information in the DRFs was filtered out. In this work, we explored the use of such intensity information for potential improvement in WEPL accuracy and imaging quality. Three WEPL derivation methods based on, respectively, the RMS only, the intensity only, and the intensity-weighted RMS were tested and compared in terms of the quality of obtained radiograph images and the accuracy of WEPL values. A Gammex CT calibration phantom containing inserts made of various tissue substitute materials with independently measured relative stopping powers (RSP) was used to assess the imaging performances. Improved image quality with enhanced interfaces was achieved while preserving the accuracy by using intensity information in the calibration. Other objects, including an anthropomorphic head phantom, a proton therapy range compensator, a frozen lamb’s head and an ‘image quality phantom’ were also imaged. Both the RMS only and the intensity-weighted RMS methods derived RSPs within  ±  1% for most of the Gammex phantom inserts, with a mean absolute percentage error of 0.66% for all inserts. In the case of the insert with a titanium rod, the method based on RMS completely failed, whereas that based on the intensity-weighted RMS was qualitatively valid. The use of intensity greatly enhanced the interfaces between different materials in the obtained WEPL images, suggesting the potential for image guidance in areas such as patient positioning and tumor tracking by proton

  6. A nucleator arms race: cellular control of actin assembly.

    Science.gov (United States)

    Campellone, Kenneth G; Welch, Matthew D

    2010-04-01

    For over a decade, the actin-related protein 2/3 (ARP2/3) complex, a handful of nucleation-promoting factors and formins were the only molecules known to directly nucleate actin filament formation de novo. However, the past several years have seen a surge in the discovery of mammalian proteins with roles in actin nucleation and dynamics. Newly recognized nucleation-promoting factors, such as WASP and SCAR homologue (WASH), WASP homologue associated with actin, membranes and microtubules (WHAMM), and junction-mediating regulatory protein (JMY), stimulate ARP2/3 activity at distinct cellular locations. Formin nucleators with additional biochemical and cellular activities have also been uncovered. Finally, the Spire, cordon-bleu and leiomodin nucleators have revealed new ways of overcoming the kinetic barriers to actin polymerization.

  7. International Conference Intergranular and Interphase Boundaries (9th) (IIB󈨦) Held in Prague, Czech Republic, 6 - 9 July 1998

    Science.gov (United States)

    1998-07-09

    highly resistive to void swelling. According to positron annihilation studies [1] the onset of vacancy long-range mi- gration is almost not affected...transformations in the chemical composition of the dispersoids. Two different types of the TiO2 particles depending on Cr behaviour are found to form...in the steel in the course of annealing. In the first case, Cr enriches mainly in nearest subsurface layers of the TiO2 dispersoids. In the second

  8. Symposium Q: Magnetic Thin Films, Heterostructures, and Device Materials

    Science.gov (United States)

    2007-05-22

    ing, and applications. version, hydrogen, and nuclear research. Examples of extended functionality of In addition to the presentations on government...and the pure Fe is bcc phase. United States Department of Energyi National Nuclear Security From these evidences, it indicates that the laminates form...in the regions where the self assembled monolayer is polar in Escudero3; ’Centro de Investigacion en Quimica Aplicada, Saltillo, nature, and it is non

  9. Technologie výroby a kontrola jakosti živočišných produktů s označením zaručená tradiční specialita

    OpenAIRE

    Chovancová, Tereza

    2015-01-01

    This bachelor thesis deals with the issues concerned with the performance, production and quality control of the meat products with "Traditional specialities guaranteed" indication. The first part deals with historical development of producing and range of meat products in the Czech Republic, the range of meat products according to the ordinance 326/2001 Sb. as amended. Moreover, the basic methods of meat products manufacturing are described (resources, pulverization, mixing, stuffing, formin...

  10. The structure of FMNL2-Cdc42 yields insights into the mechanism of lamellipodia and filopodia formation

    Science.gov (United States)

    Kühn, Sonja; Erdmann, Constanze; Kage, Frieda; Block, Jennifer; Schwenkmezger, Lisa; Steffen, Anika; Rottner, Klemens; Geyer, Matthias

    2015-05-01

    Formins are actin polymerization factors that elongate unbranched actin filaments at the barbed end. Rho family GTPases activate Diaphanous-related formins through the relief of an autoregulatory interaction. The crystal structures of the N-terminal domains of human FMNL1 and FMNL2 in complex with active Cdc42 show that Cdc42 mediates contacts with all five armadillo repeats of the formin with specific interactions formed by the Rho-GTPase insert helix. Mutation of three residues within Rac1 results in a gain-of-function mutation for FMNL2 binding and reconstitution of the Cdc42 phenotype in vivo. Dimerization of FMNL1 through a parallel coiled coil segment leads to formation of an umbrella-shaped structure that--together with Cdc42--spans more than 15 nm in diameter. The two interacting FMNL-Cdc42 heterodimers expose six membrane interaction motifs on a convex protein surface, the assembly of which may facilitate actin filament elongation at the leading edge of lamellipodia and filopodia.

  11. Personal reflections on the highlights and changes in radiation and radioisotope measurement applications

    International Nuclear Information System (INIS)

    Gardner, Robin P.; Lee, Kyoung O.

    2015-01-01

    This paper describes the recent changes that the authors have perceived in the use of radiation and radioisotope measurement applications. The first change is that due to the increased use of Monte Carlo simulation which has occurred from a normal evolutionary process. This is due in large part to the increased accuracy that is being obtained by the use of detector response functions (DRFs) and the simultaneous increased computational efficiency that has become available with these DRFs, the availability of a greatly improved weight windows variance reduction method, and the availability of inexpensive computer clusters. This first change is a happy one. The other change that is occurring is in response to recent terrorist activities. That change is the replacement or major change in the use of long-lived radioisotopes in radioisotope measurement and other radioisotope source applications. In general this can be done by improving the security of these radioisotope sources or by replacing them altogether by using machine sources of radiation. In either case one would like to preclude altogether or at least minimize the possibility of terrorists being able to obtain radioisotopes and use them for clandestine purposes. - Highlights: • Recent changes in radioisotope measurement applications. • Improvements in Monte Carlo simulation for treating radioisotope measurement applications. • Replacement of radioisotope sources with machine sources of radiation.

  12. Eye dosimetry and protective eye wear for interventional clinicians

    International Nuclear Information System (INIS)

    Martin, C.J.; Magee, J.S.; Sandblom; Almen, A.; Lundh, C.

    2015-01-01

    Doses to the eyes of interventional clinicians can exceed 20 mSv. Various protective devices can afford protection to the eyes with the final barrier being protective eye wear. The protection provided by lead glasses is difficult to quantify, and the majority of dosimeters are not designed to be worn under lead glasses. This study has measured dose reduction factors (DRFs) equal to the ratio of the dose with no protection, divided by that when lead glasses are worn. Glasses have been tested in X-ray fields using anthropomorphic phantoms to simulate the patient and clinician. DRFs for X-rays incident from the front vary from 5.2 to 7.6, while values for orientations reminiscent of clinical practice are between 1.4 and 5.2. Results suggest that a DRF of two is a conservative factor that could be applied to personal dosimeter measurements to account for the dose reduction provided by most types of lead glasses. (authors)

  13. Should Women Younger Than 40 Years of Age With Invasive Breast Cancer Have a Mastectomy?: 15-Year Outcomes in a Population-Based Cohort

    International Nuclear Information System (INIS)

    Cao, Jeffrey Q.; Truong, Pauline T.; Olivotto, Ivo A.; Olson, Robert; Coulombe, Genevieve; Keyes, Mira; Weir, Lorna; Gelmon, Karen; Bernstein, Vanessa; Woods, Ryan; Speers, Caroline; Tyldesley, Scott

    2014-01-01

    Purpose: Optimal local management for young women with early-stage breast cancer remains controversial. This study examined 15-year outcomes among women younger than 40 years treated with breast-conserving surgery plus whole-breast radiation therapy (BCT) compared with those treated with modified radical mastectomy (MRM). Methods and Materials: Women aged 20 to 39 years with early-stage breast cancer diagnosed between 1989 and 2003 were identified in a population-based database. Primary outcomes of breast cancer–specific survival (BCSS), overall survival (OS) and secondary outcomes of local relapse–free survival (LRFS), locoregional relapse–free survival (LRRFS), and distant relapse–free survival (DRFS) were calculated using Kaplan-Meier methods and compared between BCT and MRM cohorts using log-rank tests. A planned subgroup analysis was performed on patients considered “ideal” for BCT (ie, T1N0, negative margins and no extensive ductal carcinoma in situ) and in whom local therapy may have the largest impact on survival because of low systemic risk. Results: 965 patients were identified; 616 had BCT and 349 had MRM. The median follow-up time was 14.4 years (range, 8.4-23.3 years). Overall, 15-year rates of BCSS (76.0% vs 74.1%, P=.62), OS (74.2% vs 73.0%, P=.75), LRFS (85.4% vs 86.5%, P=.95), LRRFS (82.2% vs 81.6%, P=.61), and DRFS (74.4% vs 71.6%, P=.40) were similar between the BCT and MRM cohorts. In the “ideal” for BCT subgroup, there were 219 BCT and 67 MRM patients with a median follow-up time of 15.5 years. The 15-year BCSS (86.1% vs 82.9%, P=.57), OS (82.6% vs 82.9%, P=.89), LRFS (86.2% vs 84.2%, P=.50), LRRFS (83.1% vs 78.3%, P=.24), and DRFS (84.8% vs 79.1%, P=.17) were similar in the BCT and MRM cohorts. Conclusions: This population-based analysis with long-term follow-up confirmed that women younger than 40 years treated with BCT had similar 15-year outcomes compared with MRM. Young age alone is not a contraindication to BCT

  14. AcEST: DK958823 [AcEST

    Lifescience Database Archive (English)

    Full Text Available MPPLEG 2401 >sp|Q8AZM0|POLS_BSNV Structural polyprotein OS=Blotched snakehead virus PE=1 SV=1 Length = 1069 ...p|Q68749|POLG_HCVBB Genome polyprotein OS=Hepatitis C virus gen... 33 0.98 sp|Q8AZM0|POLS_BSNV Structural polyprotein OS=Blotched sna...kehead... 31 4.8 sp|Q7XWS7|FH12_ORYSJ Formin-like protein 12 OS=Oryza sativa subs..

  15. A Bipolar Spindle of Antiparallel ParM Filaments Drives Bacterial Plasmid Segregation

    DEFF Research Database (Denmark)

    Gayathri, P; Fujii, T; Møller-Jensen, Jakob

    2012-01-01

    the spindle between ParRC complexes on sister plasmids. Using a combination of structural work and total internal reflection fluorescence microscopy, we show that ParRC bound and could accelerate growth at only one end of polar ParM filaments, mechanistically resembling eukaryotic formins. The architecture...... of ParM filaments enabled two ParRC-bound filaments to associate in an antiparallel orientation, forming a bipolar spindle. The spindle elongated as a bundle of at least two antiparallel filaments, thereby pushing two plasmid clusters toward the poles....

  16. WE-EF-303-10: Single- Detector Proton Radiography as a Portal Imaging Equivalent for Proton Therapy

    Energy Technology Data Exchange (ETDEWEB)

    Doolan, P [University College London Hospital, London (United Kingdom); Bentefour, E [Ion Beam Applications, Louvain-la-Neuve (Belgium); Testa, M; Cascio, E; Lu, H [Massachussetts General Hospital, Boston, MA (United States); Royle, G [University College London, London (United Kingdom); Gottschalk, B [Harvard University, Cambridge, MA (United States)

    2015-06-15

    Purpose: In proton therapy, patient alignment is of critical importance due to the sensitivity of the proton range to tissue heterogeneities. Traditionally proton radiography is used for verification of the water-equivalent path length (WEPL), which dictates the depth protons reach. In this work we propose its use for alignment. Additionally, many new proton centers have cone-beam computed tomography in place of beamline X-ray imaging and so proton radiography offers a unique patient alignment verification similar to portal imaging in photon therapy. Method: Proton radiographs of a CIRS head phantom were acquired using the Beam Imaging System (BIS) (IBA, Louvain-la-Neuve) in a horizontal beamline. A scattered beam was produced using a small, dedicated, range modulator (RM) wheel fabricated out of aluminum. The RM wheel was rotated slowly (20 sec/rev) using a stepper motor to compensate for the frame rate of the BIS (120 ms). Dose rate functions (DRFs) over two RM wheel rotations were acquired. Calibration was made with known thicknesses of homogeneous solid water. For each pixel the time width, skewness and kurtosis of the DRFs were computed. The time width was used to compute the object WEPL. In the heterogeneous phantom, the excess skewness and excess kurtosis (i.e. difference from homogeneous cases) were computed and assessed for suitability for patient set up. Results: The technique allowed for the simultaneous production of images that can be used for WEPL verification, showing few internal details, and excess skewness and kurtosis images that can be used for soft tissue alignment. These latter images highlight areas where range mixing has occurred, correlating with phantom heterogeneities. Conclusion: The excess skewness and kurtosis images contain details that are not visible in the WET images. These images, unique to the time-resolved proton radiographic method, could be used for patient set up according to soft tissues.

  17. Response functions for computing absorbed dose to skeletal tissues from photon irradiation-an update

    Energy Technology Data Exchange (ETDEWEB)

    Johnson, Perry B; Bahadori, Amir A [Nuclear and Radiological Engineering, University of Florida, Gainesville, FL 32611 (United States); Eckerman, Keith F [Life Sciences Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831 (United States); Lee, Choonsik [Radiation Epidemiology Branch, National Cancer Institute, Bethesda, MD 20892 (United States); Bolch, Wesley E, E-mail: wbolch@ufl.edu [Nuclear and Radiological/Biomedical Engineering, University of Florida, Gainesville, FL 32611 (United States)

    2011-04-21

    A comprehensive set of photon fluence-to-dose response functions (DRFs) is presented for two radiosensitive skeletal tissues-active and total shallow marrow-within 15 and 32 bone sites, respectively, of the ICRP reference adult male. The functions were developed using fractional skeletal masses and associated electron-absorbed fractions as reported for the UF hybrid adult male phantom, which in turn is based upon micro-CT images of trabecular spongiosa taken from a 40 year male cadaver. The new DRFs expand upon both the original set of seven functions produced in 1985, and a 2007 update calculated under the assumption of secondary electron escape from spongiosa. In this study, it is assumed that photon irradiation of the skeleton will yield charged particle equilibrium across all spongiosa regions at energies exceeding 200 keV. Kerma coefficients for active marrow, inactive marrow, trabecular bone and spongiosa at higher energies are calculated using the DRF algorithm setting the electron-absorbed fraction for self-irradiation to unity. By comparing kerma coefficients and DRF functions, dose enhancement factors and mass energy-absorption coefficient (MEAC) ratios for active marrow to spongiosa were derived. These MEAC ratios compared well with those provided by the NIST Physical Reference Data Library (mean difference of 0.8%), and the dose enhancement factors for active marrow compared favorably with values calculated in the well-known study published by King and Spiers (1985 Br. J. Radiol. 58 345-56) (mean absolute difference of 1.9 percentage points). Additionally, dose enhancement factors for active marrow were shown to correlate well with the shallow marrow volume fraction (R{sup 2} = 0.91). Dose enhancement factors for the total shallow marrow were also calculated for 32 bone sites representing the first such derivation for this target tissue.

  18. Wnt pathway reprogramming during human embryonal carcinoma differentiation and potential for therapeutic targeting

    International Nuclear Information System (INIS)

    Snow, Grace E; Kasper, Allison C; Busch, Alexander M; Schwarz, Elisabeth; Ewings, Katherine E; Bee, Thomas; Spinella, Michael J; Dmitrovsky, Ethan; Freemantle, Sarah J

    2009-01-01

    Testicular germ cell tumors (TGCTs) are classified as seminonas or non-seminomas of which a major subset is embryonal carcinoma (EC) that can differentiate into diverse tissues. The pluripotent nature of human ECs resembles that of embryonic stem (ES) cells. Many Wnt signalling species are regulated during differentiation of TGCT-derived EC cells. This study comprehensively investigated expression profiles of Wnt signalling components regulated during induced differentiation of EC cells and explored the role of key components in maintaining pluripotency. Human embryonal carcinoma cells were stably infected with a lentiviral construct carrying a canonical Wnt responsive reporter to assess Wnt signalling activity following induced differentiation. Cells were differentiated with all-trans retinoic acid (RA) or by targeted repression of pluripotency factor, POU5F1. A Wnt pathway real-time-PCR array was used to evaluate changes in gene expression as cells differentiated. Highlighted Wnt pathway genes were then specifically repressed using siRNA or stable shRNA and transfected EC cells were assessed for proliferation, differentiation status and levels of core pluripotency genes. Canonical Wnt signalling activity was low basally in undifferentiated EC cells, but substantially increased with induced differentiation. Wnt pathway gene expression levels were compared during induced differentiation and many components were altered including ligands (WNT2B), receptors (FZD5, FZD6, FZD10), secreted inhibitors (SFRP4, SFRP1), and other effectors of Wnt signalling (FRAT2, DAAM1, PITX2, Porcupine). Independent repression of FZD5, FZD7 and WNT5A using transient as well as stable methods of RNA interference (RNAi) inhibited cell growth of pluripotent NT2/D1 human EC cells, but did not appreciably induce differentiation or repress key pluripotency genes. Silencing of FZD7 gave the greatest growth suppression in all human EC cell lines tested including NT2/D1, NT2/D1-R1, Tera-1 and 833

  19. INSIGHTS INTO THE MECHANICS OF CYTOKINETIC RING ASSEMBLY USING 3D MODELING.

    Science.gov (United States)

    Bidone, Tamara Carla; Tang, Haosu; Vavylonis, Dimitrios

    During fission yeast cytokinesis, actin filaments nucleated by cortical formin Cdc12 are captured by myosin motors bound to a band of cortical nodes. The myosin motors exert forces that pull nodes together into a contractile ring. Cross-linking interactions help align actin filaments and nodes into a single bundle. Mutations in the myosin motor domain and changes in the concentration of cross-linkers alpha-actinin and fimbrin alter the morphology of the condensing network, leading to clumps, rings or extended meshworks. How the contractile tension developing during ring formation depends on the interplay between network morphology, myosin motor activity, cross-linking and actin filament turnover remains to be elucidated. We addressed this question using a 3D computational model in which semiflexible actin filaments (represented as beads connected by springs) grow from formins, can be captured by myosin in neighboring nodes, and get cross-linked with one another through an attractive interaction. We identify regimes of tension generation between connected nodes under a wide set of conditions regarding myosin dynamics and strength of cross-linking between actin filaments. We find conditions that maximize circumferential tension, correlate them with network morphology and propose experiments to test these predictions. This work addresses "Morphogenesis of soft and living matter" using computational modeling to simulate cytokinetic ring assembly from the key molecular mechanisms of viscoelastic cross-linked actin networks that include active molecular motors.

  20. MiR-33a Decreases High-Density Lipoprotein-Induced Radiation Sensitivity in Breast Cancer

    Energy Technology Data Exchange (ETDEWEB)

    Wolfe, Adam R.; Bambhroliya, Arvind [Department of Radiation Oncology, The University of Texas MD Anderson Cancer Center, Houston, Texas (United States); Morgan Welch Inflammatory Breast Cancer Research Program and Clinic, The University of Texas MD Anderson Cancer Center, Houston, Texas (United States); Reddy, Jay P. [Department of Radiation Oncology, The University of Texas MD Anderson Cancer Center, Houston, Texas (United States); Debeb, Bisrat G. [Department of Radiation Oncology, The University of Texas MD Anderson Cancer Center, Houston, Texas (United States); Morgan Welch Inflammatory Breast Cancer Research Program and Clinic, The University of Texas MD Anderson Cancer Center, Houston, Texas (United States); Huo, Lei [Department of Pathology, The University of Texas MD Anderson Cancer Center, Houston, Texas (United States); Larson, Richard; Li, Li [Department of Radiation Oncology, The University of Texas MD Anderson Cancer Center, Houston, Texas (United States); Morgan Welch Inflammatory Breast Cancer Research Program and Clinic, The University of Texas MD Anderson Cancer Center, Houston, Texas (United States); Ueno, Naoto T. [Morgan Welch Inflammatory Breast Cancer Research Program and Clinic, The University of Texas MD Anderson Cancer Center, Houston, Texas (United States); Woodward, Wendy A., E-mail: wwoodward@mdanderson.org [Department of Radiation Oncology, The University of Texas MD Anderson Cancer Center, Houston, Texas (United States); Morgan Welch Inflammatory Breast Cancer Research Program and Clinic, The University of Texas MD Anderson Cancer Center, Houston, Texas (United States)

    2016-06-01

    Purpose: We previously showed that high-density lipoprotein (HDL) radiosensitizes inflammatory breast cancer (IBC) cells in vitro and is associated with better local control after radiation therapy in IBC patients. The microRNA miR-33 family negatively regulates the adenosine triphosphate binding cassette transporter subfamily A member 1. We hypothesized that variations in miR-33a expression in IBC cancer cells versus non-IBC cells would correlate with radiation sensitivity following exposure to HDL in vitro. Methods and Materials: MiR-33a expression was analyzed by reverse transcriptase–polymerase chain reaction in 4 cell lines representing common clinical breast cancer subtypes. Overexpression and knockdown of miR-33a was demonstrated via transfection of an miR-33a mimic or an anti-miR-33a construct in high- and low-expressing miR-33a cell lines. Clonogenic survival in vitro in these cells was quantified at baseline and following HDL treatment. MiR-33a expression on distant relapse-free survival (DRFS) of 210 cases downloaded from the Oxford breast cancer dataset was determined. Results: Expression levels of miR-33a were lower in IBC cell lines and IBC tumor samples than in non-IBC cell lines and normal breast tissue. Cholesterol concentrations in the cell membranes were higher in IBC cells than in non-IBC cells. Clonogenic survival following 24 hours of HDL treatment was decreased in response to irradiation in the low-miR-33a–expressing cell lines SUM149 and KPL4, but survival following HDL treatment decreased in the high-miR-33a–expressing cell lines MDA-MB-231 and SUM159. In the high-miR-33a–expressing cell lines, anti-miR-33a transfection decreased radiation resistance in clonogenic assays. Conversely, in the low-miR-33a–expressing cell lines, the miR-33a mimic reversed the HDL-induced radiation sensitization. Breast cancer patients in the top quartile based on miR-33a expression had markedly lower rates of DRFS than the bottom quartile (P

  1. SU-C-207A-05: Feature Based Water Equivalent Path Length (WEPL) Determination for Proton Radiography by the Technique of Time Resolved Dose Measurement

    International Nuclear Information System (INIS)

    Zhang, R; Jee, K; Sharp, G; Flanz, J; Lu, H

    2016-01-01

    Purpose: Studies show that WEPL can be determined from modulated dose rate functions (DRF). However, the previous calibration method based on statistics of the DRF is sensitive to energy mixing of protons due to scattering through different materials (termed as range mixing here), causing inaccuracies in the determination of WEPL. This study intends to explore time-domain features of the DRF to reduce the effect of range mixing in proton radiography (pRG) by this technique. Methods: An amorphous silicon flat panel (PaxScan™ 4030CB, Varian Medical Systems, Inc., Palo Alto, CA) was placed behind phantoms to measure DRFs from a proton beam modulated by a specially designed modulator wheel. The performance of two methods, the previously used method based on the root mean square (RMS) and the new approach based on time-domain features of the DRF, are compared for retrieving WEPL and RSP from pRG of a Gammex phantom. Results: Calibration by T_8_0 (the time point for 80% of the major peak) was more robust to range mixing and produced WEPL with improved accuracy. The error of RSP was reduced from 8.2% to 1.7% for lung equivalent material, with the mean error for all other materials reduced from 1.2% to 0.7%. The mean error of the full width at half maximum (FWHM) of retrieved inserts was decreased from 25.85% to 5.89% for the RMS and T_8_0 method respectively. Monte Carlo simulations in simplified cases also demonstrated that the T_8_0 method is less sensitive to range mixing than the RMS method. Conclusion: WEPL images have been retrieved based on single flat panel measured DRFs, with inaccuracies reduced by exploiting time-domain features as the calibration parameter. The T_8_0 method is validated to be less sensitive to range mixing and can thus retrieve the WEPL values in proximity of interfaces with improved numerical and spatial accuracy for proton radiography.

  2. Distal radius fracture arthroscopic intraarticular displacement measurement after open reduction and internal fixation from a volar approach.

    Science.gov (United States)

    Ono, Hiroshi; Furuta, Kazuhiko; Fujitani, Ryotaro; Katayama, Takeshi; Akahane, Manabu

    2010-07-01

    The purpose of this study was to assess articular surface reduction arthroscopically after volar locked-plate fixation of distal radius fractures (DRFs) via fluoroscopyguided open reduction/internal fixation. We also aimed to develop preoperative radiographic criteria to help assist in determining which DRFs may need arthroscopic evaluation. A total of 31 consecutive patients with DRF were prospectively enrolled. Posteroanterior (PA) and lateral radiographs as well as axial, coronal, and sagittal CT scans were obtained just after attempted reduction of the DRF. The widest articular displacement at the radiocarpal joint surface of the distal radius (preopD) was then measured using a digital radiography imaging system. The DRF was reduced under fluoroscopy, and a volar locked plate was applied. The degree of residual articular displacement was then measured arthroscopically, and the maximum displacement (postopD) was measured with a calibrated probe. Of the 31 patients, 7 had an arthroscopically assessed maximum postopD of > or = 2 mm after internal fixation. The correlation coefficients between each preopD and postopD of all radiographs and CTs were statistically significant. The cutoff values were 0.5 mm for PA radiographs, 2.10 mm for lateral radiographs, 2.15 mm for axial CT scans, 3.15 mm for coronal CT scans, and 1.20 mm for sagittal CT scans. All cutoff values for PA and lateral radiographs and for axial, coronal, and sagittal CT scans were unsuitable as screening criteria for arthroscopic reduction of DRF because of their low sensitivities and specificities. The cutoff value of the new preopD (the sum of the preopDs determined by lateral radiography and coronal CT scan) was 5.80 mm, and its sensitivity and specificity were 100% and 83.3%, respectively. Because a new preopD cutoff value of 5.80 mm is a good indicator for residual articular displacement after internal fixation of >2 mm, it is also a good indicator for the need for arthroscopic evaluation after

  3. Distal radius fracture arthroscopic intraarticular displacement measurement after open reduction and internal fixation from a volar approach

    International Nuclear Information System (INIS)

    Ono, Hiroshi; Furuta, Kazuhiko; Fujitani, Ryotaro; Katayama, Takeshi; Akahane, Manabu

    2010-01-01

    The purpose of this study was to assess articular surface reduction arthroscopically after volar locked-plate fixation of distal radius fractures (DRFs) via fluoroscopy-guided open reduction/internal fixation. We also aimed to develop preoperative radiographic criteria to help assist in determining which DRFs may need arthroscopic evaluation. A total of 31 consecutive patients with DRF were prospectively enrolled. Posteroanterior (PA) and lateral radiographs as well as axial, coronal, and sagittal CT scans were obtained just after attempted reduction of the DRF. The widest articular displacement at the radiocarpal joint surface of the distal radius (preopD) was then measured using a digital radiography imaging system. The DRF was reduced under fluoroscopy, and a volar locked plate was applied. The degree of residual articular displacement was then measured arthroscopically, and the maximum displacement (postopD) was measured with a calibrated probe. Of the 31 patients, 7 had an arthroscopically assessed maximum postopD of ≥2 mm after internal fixation. The correlation coefficients between each preopD and postopD of all radiographs and CTs were statistically significant. The cutoff values were 0.5 mm for PA radiographs, 2.10 mm for lateral radiographs, 2.15 mm for axial CT scans, 3.15 mm for coronal CT scans, and 1.20 mm for sagittal CT scans. All cutoff values for PA and lateral radiographs and for axial, coronal, and sagittal CT scans were unsuitable as screening criteria for arthroscopic reduction of DRF because of their low sensitivities and specificities. The cutoff value of the new preopD (the sum of the preopDs determined by lateral radiography and coronal CT scan) was 5.80 mm, and its sensitivity and specificity were 100% and 83.3%, respectively. Because a new preopD cutoff value of 5.80 mm is a good indicator for residual articular displacement after internal fixation of >2 mm, it is also a good indicator for the need for arthroscopic evaluation after

  4. Response functions for computing absorbed dose to skeletal tissues from neutron irradiation

    Science.gov (United States)

    Bahadori, Amir A.; Johnson, Perry; Jokisch, Derek W.; Eckerman, Keith F.; Bolch, Wesley E.

    2011-11-01

    Spongiosa in the adult human skeleton consists of three tissues—active marrow (AM), inactive marrow (IM) and trabecularized mineral bone (TB). AM is considered to be the target tissue for assessment of both long-term leukemia risk and acute marrow toxicity following radiation exposure. The total shallow marrow (TM50), defined as all tissues lying within the first 50 µm of the bone surfaces, is considered to be the radiation target tissue of relevance for radiogenic bone cancer induction. For irradiation by sources external to the body, kerma to homogeneous spongiosa has been used as a surrogate for absorbed dose to both of these tissues, as direct dose calculations are not possible using computational phantoms with homogenized spongiosa. Recent micro-CT imaging of a 40 year old male cadaver has allowed for the accurate modeling of the fine microscopic structure of spongiosa in many regions of the adult skeleton (Hough et al 2011 Phys. Med. Biol. 56 2309-46). This microstructure, along with associated masses and tissue compositions, was used to compute specific absorbed fraction (SAF) values for protons originating in axial and appendicular bone sites (Jokisch et al 2011 Phys. Med. Biol. 56 6857-72). These proton SAFs, bone masses, tissue compositions and proton production cross sections, were subsequently used to construct neutron dose-response functions (DRFs) for both AM and TM50 targets in each bone of the reference adult male. Kerma conditions were assumed for other resultant charged particles. For comparison, AM, TM50 and spongiosa kerma coefficients were also calculated. At low incident neutron energies, AM kerma coefficients for neutrons correlate well with values of the AM DRF, while total marrow (TM) kerma coefficients correlate well with values of the TM50 DRF. At high incident neutron energies, all kerma coefficients and DRFs tend to converge as charged-particle equilibrium is established across the bone site. In the range of 10 eV to 100 Me

  5. MiR-33a Decreases High-Density Lipoprotein-Induced Radiation Sensitivity in Breast Cancer

    International Nuclear Information System (INIS)

    Wolfe, Adam R.; Bambhroliya, Arvind; Reddy, Jay P.; Debeb, Bisrat G.; Huo, Lei; Larson, Richard; Li, Li; Ueno, Naoto T.; Woodward, Wendy A.

    2016-01-01

    Purpose: We previously showed that high-density lipoprotein (HDL) radiosensitizes inflammatory breast cancer (IBC) cells in vitro and is associated with better local control after radiation therapy in IBC patients. The microRNA miR-33 family negatively regulates the adenosine triphosphate binding cassette transporter subfamily A member 1. We hypothesized that variations in miR-33a expression in IBC cancer cells versus non-IBC cells would correlate with radiation sensitivity following exposure to HDL in vitro. Methods and Materials: MiR-33a expression was analyzed by reverse transcriptase–polymerase chain reaction in 4 cell lines representing common clinical breast cancer subtypes. Overexpression and knockdown of miR-33a was demonstrated via transfection of an miR-33a mimic or an anti-miR-33a construct in high- and low-expressing miR-33a cell lines. Clonogenic survival in vitro in these cells was quantified at baseline and following HDL treatment. MiR-33a expression on distant relapse-free survival (DRFS) of 210 cases downloaded from the Oxford breast cancer dataset was determined. Results: Expression levels of miR-33a were lower in IBC cell lines and IBC tumor samples than in non-IBC cell lines and normal breast tissue. Cholesterol concentrations in the cell membranes were higher in IBC cells than in non-IBC cells. Clonogenic survival following 24 hours of HDL treatment was decreased in response to irradiation in the low-miR-33a–expressing cell lines SUM149 and KPL4, but survival following HDL treatment decreased in the high-miR-33a–expressing cell lines MDA-MB-231 and SUM159. In the high-miR-33a–expressing cell lines, anti-miR-33a transfection decreased radiation resistance in clonogenic assays. Conversely, in the low-miR-33a–expressing cell lines, the miR-33a mimic reversed the HDL-induced radiation sensitization. Breast cancer patients in the top quartile based on miR-33a expression had markedly lower rates of DRFS than the bottom quartile (P

  6. Proteomic analysis in giant axonal neuropathy: new insights into disease mechanisms.

    Science.gov (United States)

    Mussche, Silke; De Paepe, Boel; Smet, Joél; Devreese, Katrien; Lissens, Willy; Rasic, Vedrana Milic; Murnane, Matthew; Devreese, Bart; Van Coster, Rudy

    2012-08-01

    Giant axonal neuropathy (GAN) is a progressive hereditary disease that affects the peripheral and central nervous systems. It is characterized morphologically by aggregates of intermediate filaments in different tissues. Mutations have been reported in the gene that codes for gigaxonin. Nevertheless, the underlying molecular mechanism remains obscure. Cell lines from 4 GAN patients and 4 controls were analyzed by iTRAQ. Among the dysregulated proteins were ribosomal protein L29, ribosomal protein L37, galectin-1, glia-derived nexin, and aminopeptidase N. Also, nuclear proteins linked to formin-binding proteins were found to be dysregulated. Although the major role of gigaxonin is reported to be degradation of cytoskeleton-associated proteins, the amount of 76 structural cytoskeletal proteins was unaltered. Several of the dysregulated proteins play a role in cytoskeletal reorganization. Based on these findings, we speculate that disturbed cytoskeletal regulation is responsible for formation of aggregates of intermediate filaments. Copyright © 2012 Wiley Periodicals, Inc.

  7. Specific cytoarchitectureal changes in hippocampal subareas in daDREAM mice.

    Science.gov (United States)

    Mellström, Britt; Kastanauskaite, Asta; Knafo, Shira; Gonzalez, Paz; Dopazo, Xose M; Ruiz-Nuño, Ana; Jefferys, John G R; Zhuo, Min; Bliss, Tim V P; Naranjo, Jose R; DeFelipe, Javier

    2016-02-29

    Transcriptional repressor DREAM (downstream regulatory element antagonist modulator) is a Ca(2+)-binding protein that regulates Ca(2+) homeostasis through gene regulation and protein-protein interactions. It has been shown that a dominant active form (daDREAM) is implicated in learning-related synaptic plasticity such as LTP and LTD in the hippocampus. Neuronal spines are reported to play important roles in plasticity and memory. However, the possible role of DREAM in spine plasticity has not been reported. Here we show that potentiating DREAM activity, by overexpressing daDREAM, reduced dendritic basal arborization and spine density in CA1 pyramidal neurons and increased spine density in dendrites in dentate gyrus granule cells. These microanatomical changes are accompanied by significant modifications in the expression of specific genes encoding the cytoskeletal proteins Arc, Formin 1 and Gelsolin in daDREAM hippocampus. Our results strongly suggest that DREAM plays an important role in structural plasticity in the hippocampus.

  8. Coordination by Cdc42 of Actin, Contractility, and Adhesion for Melanoblast Movement in Mouse Skin

    DEFF Research Database (Denmark)

    Woodham, Emma F; Paul, Nikki R; Tyrrell, Benjamin

    2017-01-01

    traverse the dermis to reach the epidermis of the skin and hair follicles. We previously established that Rac1 signals via Scar/WAVE and Arp2/3 to effect pseudopod extension and migration of melanoblasts in skin. Here we show that RhoA is redundant in the melanocyte lineage but that Cdc42 coordinates...... multiple motility systems independent of Rac1. Similar to Rac1 knockouts, Cdc42 null mice displayed a severe loss of pigmentation, and melanoblasts showed cell-cycle progression, migration, and cytokinesis defects. However, unlike Rac1 knockouts, Cdc42 null melanoblasts were elongated and displayed large...... null cells lacked the ability to polarize their Golgi and coordinate motility systems for efficient movement. Loss of Cdc42 de-coupled three main systems: actin assembly via the formin FMNL2 and Arp2/3, active myosin-II localization, and integrin-based adhesion dynamics....

  9. RhoA determines disease progression by controlling neutrophil motility and restricting hyperresponsiveness

    DEFF Research Database (Denmark)

    Jennings, Richard T; Strengert, Monika; Hayes, Patti

    2014-01-01

    Neutrophil responses are central to host protection and inflammation. Neutrophil activation follows a two-step process where priming amplifies responses to activating stimuli. Priming is essential for life span extension, chemotaxis and respiratory burst activity. Here we show that the cytoskeletal...... organizer RhoA suppresses neutrophil priming via formins. Premature granule exocytosis in Rho-deficient neutrophils activated numerous signaling pathways and amplified superoxide generation. Deletion of Rho altered front-to-back coordination by simultaneously increasing uropod elongation, leading edge...... neutrophils exacerbated LPS-mediated lung injury, deleting Rho in innate immune cells was highly protective in Influenza A virus infection. Hence, Rho is a key regulator of disease progression by maintaining neutrophil quiescence and suppressing hyperresponsiveness....

  10. mDia2 and CXCL12/CXCR4 chemokine signaling intersect to drive tumor cell amoeboid morphological transitions.

    Science.gov (United States)

    Wyse, Meghan M; Goicoechea, Silvia; Garcia-Mata, Rafael; Nestor-Kalinoski, Andrea L; Eisenmann, Kathryn M

    2017-03-04

    Morphological plasticity in response to environmental cues in migrating cancer cells requires F-actin cytoskeletal rearrangements. Conserved formin family proteins play critical roles in cell shape, tumor cell motility, invasion and metastasis, in part, through assembly of non-branched actin filaments. Diaphanous-related formin-2 (mDia2/Diaph3/Drf3/Dia) regulates mesenchymal-to-amoeboid morphological conversions and non-apoptotic blebbing in tumor cells by interacting with its inhibitor diaphanous-interacting protein (DIP), and disrupting cortical F-actin assembly and bundling. F-actin disruption is initiated by a CXCL12-dependent mechanism. Downstream CXCL12 signaling partners inducing mDia2-dependent amoeboid conversions remain enigmatic. We found in MDA-MB-231 tumor cells CXCL12 induces DIP and mDia2 interaction in blebs, and engages its receptor CXCR4 to induce RhoA-dependent blebbing. mDia2 and CXCR4 associate in blebs upon CXCL12 stimulation. Both CXCR4 and RhoA are required for CXCL12-induced blebbing. Neither CXCR7 nor other Rho GTPases that activate mDia2 are required for CXCL12-induced blebbing. The Rho Guanine Nucleotide Exchange Factor (GEF) Net1 is required for CXCL12-driven RhoA activation and subsequent blebbing. These results reveal CXCL12 signaling, through CXCR4, directs a Net1/RhoA/mDia-dependent signaling hub to drive cytoskeleton rearrangements to regulate morphological plasticity in tumor cells. These signaling hubs may be conserved during normal and cancer cells responding to chemotactic cues. Copyright © 2017 Elsevier Inc. All rights reserved.

  11. A mechanism of leading-edge protrusion in the absence of Arp2/3 complex.

    Science.gov (United States)

    Suraneni, Praveen; Fogelson, Ben; Rubinstein, Boris; Noguera, Philippe; Volkmann, Niels; Hanein, Dorit; Mogilner, Alex; Li, Rong

    2015-03-01

    Cells employ protrusive leading edges to navigate and promote their migration in diverse physiological environments. Classical models of leading-edge protrusion rely on a treadmilling dendritic actin network that undergoes continuous assembly nucleated by the Arp2/3 complex, forming ruffling lamellipodia. Recent work demonstrated, however, that, in the absence of the Arp2/3 complex, fibroblast cells adopt a leading edge with filopodia-like protrusions (FLPs) and maintain an ability to move, albeit with altered responses to different environmental signals. We show that formin-family actin nucleators are required for the extension of FLPs but are insufficient to produce a continuous leading edge in fibroblasts lacking Arp2/3 complex. Myosin II is concentrated in arc-like regions of the leading edge in between FLPs, and its activity is required for coordinated advancement of these regions with formin-generated FLPs. We propose that actomyosin contraction acting against membrane tension advances the web of arcs between FLPs. Predictions of this model are verified experimentally. The dependence of myosin II in leading-edge advancement helps explain the previously reported defect in directional movement in the Arpc3-null fibroblasts. We provide further evidence that this defect is cell autonomous during chemotaxis. © 2015 Suraneni et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

  12. AO Distal Radius Fracture Classification: Global Perspective on Observer Agreement

    Science.gov (United States)

    Jayakumar, Prakash; Teunis, Teun; Giménez, Beatriz Bravo; Verstreken, Frederik; Di Mascio, Livio; Jupiter, Jesse B.

    2016-01-01

    Background The primary objective of this study was to test interobserver reliability when classifying fractures by consensus by AO types and groups among a large international group of surgeons. Secondarily, we assessed the difference in inter- and intraobserver agreement of the AO classification in relation to geographical location, level of training, and subspecialty. Methods A randomized set of radiographic and computed tomographic images from a consecutive series of 96 distal radius fractures (DRFs), treated between October 2010 and April 2013, was classified using an electronic web-based portal by an invited group of participants on two occasions. Results Interobserver reliability was substantial when classifying AO type A fractures but fair and moderate for type B and C fractures, respectively. No difference was observed by location, except for an apparent difference between participants from India and Australia classifying type B fractures. No statistically significant associations were observed comparing interobserver agreement by level of training and no differences were shown comparing subspecialties. Intra-rater reproducibility was “substantial” for fracture types and “fair” for fracture groups with no difference accounting for location, training level, or specialty. Conclusion Improved definition of reliability and reproducibility of this classification may be achieved using large international groups of raters, empowering decision making on which system to utilize. Level of Evidence Level III PMID:28119795

  13. Comparative study of the radioprotective effects of cysteamine, WR-2721, and WR-1065 in cultured human cells

    International Nuclear Information System (INIS)

    Purdie, J.W.

    1979-01-01

    The compounds, 2-mercaptoethylamine (cysteamine), S-2-(3-aminopropylamino)ethylphosphorothioic acid (WR-2721), and N-(2-mercaptoethyl)-1,3-diaminopropane (WR-1065) were tested for toxicity and radioprotective effect in cultured human cells using viability (reproductive death) as an indicator. WR-2721 and WR-1065 had no harmful effect at concentrations of 4 and 10 mM for periods up to 3 hr whereas cysteamine was toxic if left in contact with the cells for more than 30 min. The response of the cells to radiation was measured after a 30-min treatment with 4 mM of each protective agent. Survival curves were constructed and the following dose reduction factors (DRFs) were calculated: WR-2721, 1.3; cysteamine, 2.3; WR-1065, 2.9. Increasing the concentration of protective agent did not increase the DRF under these conditions except in the case of cysteamine. Extending the period of treatment with drug before irradiation increased the DRF for WR-2721 and WR-1065; values up to 1.8 and 3.4 were obtained with 10 mM WR-2721 and WR-1065, respectively. With these two compounds, the rate of development of protection with time is almost independent of concentration until the maximum is attained; it is probably controlled by the rate of transport across the cell membrane, in the case of WR-1065, and by the rate of hydrolysis of WR-2721

  14. Imaging system models for small-bore DOI-PET scanners

    International Nuclear Information System (INIS)

    Takahashi, Hisashi; Kobayashi, Tetsuya; Yamaya, Taiga; Murayama, Hideo; Kitamura, Keishi; Hasegawa, Tomoyuki; Suga, Mikio

    2006-01-01

    Depth-of-interaction (DOI) information, which improves resolution uniformity in the field of view (FOV), is expected to lead to high-sensitivity PET scanners with small-bore detector rings. We are developing small-bore PET scanners with DOI detectors arranged in hexagonal or overlapped tetragonal patterns for small animal imaging or mammography. It is necessary to optimize the imaging system model because these scanners exhibit irregular detector sampling. In this work, we compared two imaging system models: (a) a parallel sub-LOR model in which the detector response functions (DRFs) are assumed to be uniform along the line of responses (LORs) and (b) a sub-crystal model in which each crystal is divided into a set of smaller volumes. These two models were applied to the overlapped tetragonal scanner (FOV 38.1 mm in diameter) and the hexagonal scanner (FOV 85.2 mm in diameter) simulated by GATE. We showed that the resolution non-uniformity of system model (b) was improved by 40% compared with that of system model (a) in the overlapped tetragonal scanner and that the resolution non-uniformity of system model (a) was improved by 18% compared with that of system model (b) in the hexagonal scanner. These results indicate that system model (b) should be applied to the overlapped tetragonal scanner and system model (a) should be applied to the hexagonal scanner. (author)

  15. Impacts of air pollution and climate on materials in Athens, Greece

    Science.gov (United States)

    Christodoulakis, John; Tzanis, Chris G.; Varotsos, Costas A.; Ferm, Martin; Tidblad, Johan

    2017-01-01

    For more than 10 years now the National and Kapodistrian University of Athens, Greece, has contributed to the UNECE (United Nations Economic Commission for Europe) ICP Materials (International Co-operative Programme on Effects on Materials including Historic and Cultural Monuments) programme for monitoring the corrosion/soiling levels of different kinds of materials due to environmental air-quality parameters. In this paper we present the results obtained from the analysis of observational data that were collected in Athens during the period 2003-2012. According to these results, the corrosion/soiling of the particular exposed materials tends to decrease over the years, except for the case of copper. Based on this long experimental database that is applicable to the multi-pollutant situation in the Athens basin, we present dose-response functions (DRFs) considering that dose stands for the air pollutant concentration, response for the material mass loss (normally per annum) and function, the relationship derived by the best statistical fit to the data.

  16. Consequences of an unusual flood event: case study of a drainage canal breach on a fluvial plain in NE Slovenia

    Science.gov (United States)

    Vidmar, Ines; Ambrožič, Bojan; Debeljak, Barbara; Dolžan, Erazem; Gregorin, Špela; Grom, Nina; Herman, Polona; Keršmanc, Teja; Mencin, Eva; Mernik, Natalija; Švara, Astrid; Trobec, Ana; Turnšek, Anita; Vodeb, Petra; Torkar, Anja; Brenčič, Mihael

    2013-04-01

    On November 4-6 2012 heavy precipitation resulted in floods in the middle and lower course of Drava River in NE Slovenia causing damage to many properties in the flooded area. The meteorological situation that led to consequent floods was characterized by high precipitation, fast snowmelt, SW wind and relatively high air temperature. The weather event was part of a cyclone which was spreading over the area of North, West and Central Europe in the direction of Central Europe and carried with it the passing of a cold front through Slovenia on November 4 and 5. The flood wave travelled on the Drava River from Austria to Slovenia past the 11 hydroelectric power plants after eventually moving over the Slovenian-Croatian border. The river discharge increased in the early morning of November 5 reaching 3165 m3/s. This work focuses on a single event in the Ptujsko polje where among other damage caused by the flooding, the river broke through the drainage canal of the Formin hydroelectric power plant and changed its course. The Ptujsko polje contains two fluvial terraces. In the area of Formin HPP, the lower terrace is 1.5 km wide and the surface as well as the groundwater gradient shift from west to east with the groundwater flowing parallel to the river. These characteristics contributed to the flooding and consequential breach in the embankment of the drainage canal. Several aspects of the recent floods are discussed including a critical reflection of data accessibility, possible causes and mechanisms behind it as well as the possibility of its forecasting. Synthesis of accessible data from open domain sources is performed with emphasis on geological conditions. Discharge and precipitation data from the data base of Slovenian Environment Agency are collected, reviewed and analyzed. The flood event itself is analyzed and described in detail. It is determined that the flood wave was different from the ones regulated by natural processes which points to an anthropogenic

  17. An mDia2/ROCK signaling axis regulates invasive egress from epithelial ovarian cancer spheroids.

    Science.gov (United States)

    Pettee, Krista M; Dvorak, Kaitlyn M; Nestor-Kalinoski, Andrea L; Eisenmann, Kathryn M

    2014-01-01

    Multi-cellular spheroids are enriched in ascites of epithelial ovarian cancer (OvCa) patients. They represent an invasive and chemoresistant cellular population fundamental to metastatic dissemination. The molecular mechanisms triggering single cell invasive egress from spheroids remain enigmatic. mDia formins are Rho GTPase effectors that are key regulators of F-actin cytoskeletal dynamics. We hypothesized that mDia2-driven F-actin dynamics promote single cell invasive transitions in clinically relevant three-dimensional (3D) OvCa spheroids. The current study is a dissection of the contribution of the F-actin assembly factor mDia2 formin in invasive transitions and using a clinically relevant ovarian cancer spheroid model. We show that RhoA-directed mDia2 activity is required for tight spheroid organization, and enrichment of mDia2 in the invasive cellular protrusions of collagen-embedded OVCA429 spheroids. Depleting mDia2 in ES-2 spheroids enhanced invasive dissemination of single amoeboid-shaped cells. This contrasts with spheroids treated with control siRNA, where a mesenchymal invasion program predominated. Inhibition of another RhoA effector, ROCK, had no impact on ES-2 spheroid formation but dramatically inhibited spheroid invasion through induction of a highly elongated morphology. Concurrent inhibition of ROCK and mDia2 blocked single cell invasion from ES-2 spheroids more effectively than inhibition of either protein alone, indicating that invasive egress of amoeboid cells from mDia2-depleted spheroids is ROCK-dependent. Our findings indicate that multiple GTPase effectors must be suppressed in order to fully block invasive egress from ovarian cancer spheroids. Furthermore, tightly regulated interplay between ROCK and mDia2 signaling pathways dictates the invasive capacities and the type of invasion program utilized by motile spheroid-derived ovarian cancer cells. As loss of the gene encoding mDia2, DRF3, has been linked to cancer progression and

  18. An mDia2/ROCK signaling axis regulates invasive egress from epithelial ovarian cancer spheroids.

    Directory of Open Access Journals (Sweden)

    Krista M Pettee

    Full Text Available Multi-cellular spheroids are enriched in ascites of epithelial ovarian cancer (OvCa patients. They represent an invasive and chemoresistant cellular population fundamental to metastatic dissemination. The molecular mechanisms triggering single cell invasive egress from spheroids remain enigmatic. mDia formins are Rho GTPase effectors that are key regulators of F-actin cytoskeletal dynamics. We hypothesized that mDia2-driven F-actin dynamics promote single cell invasive transitions in clinically relevant three-dimensional (3D OvCa spheroids. The current study is a dissection of the contribution of the F-actin assembly factor mDia2 formin in invasive transitions and using a clinically relevant ovarian cancer spheroid model. We show that RhoA-directed mDia2 activity is required for tight spheroid organization, and enrichment of mDia2 in the invasive cellular protrusions of collagen-embedded OVCA429 spheroids. Depleting mDia2 in ES-2 spheroids enhanced invasive dissemination of single amoeboid-shaped cells. This contrasts with spheroids treated with control siRNA, where a mesenchymal invasion program predominated. Inhibition of another RhoA effector, ROCK, had no impact on ES-2 spheroid formation but dramatically inhibited spheroid invasion through induction of a highly elongated morphology. Concurrent inhibition of ROCK and mDia2 blocked single cell invasion from ES-2 spheroids more effectively than inhibition of either protein alone, indicating that invasive egress of amoeboid cells from mDia2-depleted spheroids is ROCK-dependent. Our findings indicate that multiple GTPase effectors must be suppressed in order to fully block invasive egress from ovarian cancer spheroids. Furthermore, tightly regulated interplay between ROCK and mDia2 signaling pathways dictates the invasive capacities and the type of invasion program utilized by motile spheroid-derived ovarian cancer cells. As loss of the gene encoding mDia2, DRF3, has been linked to cancer

  19. Familial prostate cancer has a more aggressive course than sporadic prostate cancer after treatment for localized disease, mainly due to a higher rate of distant metastases

    International Nuclear Information System (INIS)

    Kupelian, Patrick A.; Klein, Eric A.; Suh, John H; Kupelian, Varant A.

    1997-01-01

    Purpose: We had already established that familial prostate cancer, defined as prostate cancer diagnosed in a father or brother, was an independent predictor of biochemical failure after treatment for localized disease. Our aim was to determine whether differences in outcome could be observed with respect to clinical failures (either local or distant) between the two forms of prostate cancer. Methods: Of the 1685 consecutive cases with localized prostate carcinoma treated between 1986 and 1996, patients with the following were excluded from the present study: no pretreatment Prostatic Specific Antigen (iPSA) level (n=54), no biopsy Gleason score (bGS) (n=25), adjuvant or neoadjuvant treatment (n=234), no available follow-up PSA level (n=30). We also excluded 617 patients who did not have a minimum of 3 years potential follow-up. The analysis was performed on 725 cases. Radiotherapy (RT) was the primary treatment in 330 patients and radical prostatectomy (RP) in 395 patients. Five percent had clinical stage T3 disease (n=37). Positive family history was defined as the presence of prostate cancer in a first degree relative (father or brother). The outcomes of interest were biochemical relapse-free survival (bRFS), clinical relapse-free survival (cRFS), local relapse-free survival (locRFS), distant relapse-free survival (dRFS). We used proportional hazards to analyze the effect of family history and other potential confounding variables (i.e. age, race, treatment modality, stage, biopsy GS, and iPSA levels) on treatment outcome. We included pathologic findings (extracapsular extension, seminal vesicle involvement, surgical margin involvement, and lymph node metastases) in a separate analysis for RP patients. Results: The median follow-up was 45 months. Eight percent of all cases (n=57) had a positive family history. The 5-year bRFS rates for patients with negative and positive family history were 54% and 38%, respectively (p<0.001). The 5-year cRFS rates for patients

  20. High fidelity remote sensing of snow properties from MODIS and the Airborne Snow Observatory: Snowflakes to Terabytes

    Science.gov (United States)

    Painter, T.; Mattmann, C. A.; Brodzik, M.; Bryant, A. C.; Goodale, C. E.; Hart, A. F.; Ramirez, P.; Rittger, K. E.; Seidel, F. C.; Zimdars, P. A.

    2012-12-01

    , robust inputs to water management models and systems of the future. In the push to better understand the physical and ecological processes of snowmelt and how they influence regional to global hydrologic and climatic cycles, these technologies and retrievals provide markedly improved detail. We have implemented a science computing facility anchored upon the open source Apache OODT data processing framework. Apache OODT provides adaptable, rapid, and effective workflow technologies that we leverage to execute 10s of thousands of MOD-DRFS and MODSCAG jobs in the Western US, Alaska, and High Asia, critical regions where snowmelt and runoff must be more accurately and precisely identified. Apache OODT also provides us data dissemination capabilities built upon the popular, open source WebDAV protocol that allow our system to disseminate over 20 TB of MOD-DRFS and MODSCAG to the decision making community. Our latest endeavor involves building out Apache OODT to support Geospatial exploration of our data, including providing a Leaflet.js based Map, Geoserver backed protocols, and seamless integration with our Apache OODT system. This framework provides the foundation for the ASO data system.

  1. Testing the validity of preventing chronic regional pain syndrome with vitamin C after distal radius fracture. [Corrected].

    Science.gov (United States)

    Malay, Sunitha; Chung, Kevin C

    2014-11-01

    The American Academy of Orthopaedic Surgeons recommends the use of vitamin C to prevent complex regional pain syndrome (CRPS) for patients with distal radius fractures (DRFs). We hypothesized that the evidence for supporting this recommendation is weak, based on epidemiological principles of association and causality. The specific aim of this project was to test the validity of this recommendation. We conducted a literature review to retrieve articles reporting on the use of vitamin C to prevent CRPS. Data collected included sample size, study design type, dose of vitamin C used, and outcome measures of association expressed as relative risk (RR) and odds ratio. We then applied Hill criteria to evaluate the relationship between vitamin C and CRPS. We obtained 225 articles from the database search. After the exclusion of duplicates, unrelated articles, editorial letters, and commentaries, we found 4 articles and 1 systematic review relevant to our topic. Six of the 9 Hill criteria were met, and an earlier meta-analysis showed a quantified reduction in CRPS risk. However, criteria like biological plausibility, specificity, and coherence were not met. The number of causal/association criteria met was adequate to support the scientific premise of the effect of vitamin C in preventing CRPS after DRF. Furthermore, vitamin C administration is of relatively low cost and has few complications unless administered in large doses. Owing to sufficient epidemiological evidence availability, the American Academy of Orthopaedic Surgeons recommendation of vitamin C to prevent CRPS has practical merit. Therapeutic II. Copyright © 2014 American Society for Surgery of the Hand. Published by Elsevier Inc. All rights reserved.

  2. Resting brain activity varies with dream recall frequency between subjects.

    Science.gov (United States)

    Eichenlaub, Jean-Baptiste; Nicolas, Alain; Daltrozzo, Jérôme; Redouté, Jérôme; Costes, Nicolas; Ruby, Perrine

    2014-06-01

    Dreaming is still poorly understood. Notably, its cerebral underpinning remains unclear. Neuropsychological studies have shown that lesions in the temporoparietal junction (TPJ) and/or the white matter of the medial prefrontal cortex (MPFC) lead to the global cessation of dream reports, suggesting that these regions of the default mode network have key roles in the dreaming process (forebrain 'dream-on' hypothesis). To test this hypothesis, we measured regional cerebral blood flow (rCBF) using [(15)O]H2O positron emission tomography in healthy subjects with high and low dream recall frequencies (DRFs) during wakefulness (rest) and sleep (rapid eye movement (REM) sleep, N2, and N3). Compared with Low recallers (0.5 ± 0.3 dream recall per week in average), High recallers (5.2 ± 1.4) showed higher rCBF in the TPJ during REM sleep, N3, and wakefulness, and in the MPFC during REM sleep and wakefulness. We demonstrate that the resting states of High recallers and Low recallers differ during sleep and wakefulness. It coheres with previous ERP results and confirms that a high/low DRF is associated with a specific functional organization of the brain. These results support the forebrain 'dream-on' hypothesis and suggest that TPJ and MPFC are not only involved in dream recall during wakefulness but also have a role in dreaming during sleep (production and/or encoding). Increased activity in the TPJ and MPFC might promote the mental imagery and/or memory encoding of dreams. Notably, increased activity in TPJ might facilitate attention orienting toward external stimuli and promote intrasleep wakefulness, facilitating the encoding of the dreams in memory.

  3. Recent advances in the use of ASEDRA in post processing scintillator spectra for resolution enhancement

    International Nuclear Information System (INIS)

    Sjoden, G.E.

    2012-01-01

    The ASEDRA (Advanced Synthetically Enhanced Detector Resolution Algorithm, patent pending) has been successfully applied as a post processing algorithm to both sodium iodide (NaI(Tl)) and cesium iodide (CsI(Na)) scintillator detectors to synthetically enhance their realized spectral data resolution by as much as a factor of three, wherein from these detectors the 'raw' unprocessed spectra are traditionally of poor resolution. ASEDRA uses noise reduction and built-in high resolution Monte Carlo radiation transport based detector response functions (DRFs) to rapidly post-process a spectrum in a few seconds on a standard laptop; gamma lines are extracted with an accuracy that makes the scintillator detectors competitive with higher resolution, higher material cost detectors. ASEDRA differs from other tools in the field, such as Sandia's GADRAS software, in that ASEDRA performs a differential spectrum attribution and cumulative extraction from the sample spectrum, rather than an integral-based approach, as in GADRAS. Previous publications have highlighted the successful application of ASEDRA in samples with plutonium and various isotopes. A new SmartID nuclide identification package to accompany ASEDRA has recently been implemented for test and evaluation purposes for sample attribution; in addition, the application of ASEDRA+SmartID has occurred with success in long dwell cargo monitoring and SNM detection applications, enabling new protocols for HEU detection. Overall, this paper presents recent developments and results along with a discussion of follow-on steps in the development of ASEDRA as an effective field gamma spectrum analysis tool for low cost scintillators. (author)

  4. Epidemiology of distal radius fractures in polytrauma patients and the influence of high traumatic energy transfer.

    Science.gov (United States)

    Ferree, Steven; van der Vliet, Quirine M J; Nawijn, Femke; Bhashyam, Abhiram R; Houwert, Roderick M; Leenen, Luke P H; Hietbrink, Falco

    2018-03-01

    For several extremity fractures differences in morphology, incidence rate and functional outcome were found when polytrauma patients were compared to patients with an isolated injury. This is not proven for distal radius fractures (DRF). Therefore, this study aimed to analyse fracture morphology in relation to energy transfer in both poly- and mono-trauma patients with a DRF. This was a retrospective cohort study. All patients aged 16 years and older with a DRF were included. Patients with an Injury Severity Score of 16 or higher were classified as polytrauma patients. Injuries were defined as high or low energy. All DRFs were classified using the AO/OTA fracture classification system. A total of 830 patients with a DRF were included, 12% were polytrauma. The incidence rate of DRF in polytrauma patients was 3.5%. Ipsilateral upper extremity injury was found in >30% of polytrauma and high-energy monotrauma patients, compared to 5% in low-energy monotrauma patients. More type C DRF were found in polytrauma and high-energy monotrauma patients versus low-energy monotrauma patients. Operative intervention rates for all types of DRF were similar for polytrauma and high-energy monotrauma patients. Non-union rates were higher in polytrauma patients. Higher energy mechanisms of injury, in polytrauma and high-energy monotrauma patients, were associated with more severe complex articular distal radius fractures and more ipsilateral upper extremity injuries. Polytrauma and high-energy monotrauma patient have a similar fracture morphology. However, polytrauma patients have in addition to more injured body regions also more non-union related interventions than high-energy monotrauma patients. Copyright © 2018 Elsevier Ltd. All rights reserved.

  5. Discriminative Random Field Models for Subsurface Contamination Uncertainty Quantification

    Science.gov (United States)

    Arshadi, M.; Abriola, L. M.; Miller, E. L.; De Paolis Kaluza, C.

    2017-12-01

    Application of flow and transport simulators for prediction of the release, entrapment, and persistence of dense non-aqueous phase liquids (DNAPLs) and associated contaminant plumes is a computationally intensive process that requires specification of a large number of material properties and hydrologic/chemical parameters. Given its computational burden, this direct simulation approach is particularly ill-suited for quantifying both the expected performance and uncertainty associated with candidate remediation strategies under real field conditions. Prediction uncertainties primarily arise from limited information about contaminant mass distributions, as well as the spatial distribution of subsurface hydrologic properties. Application of direct simulation to quantify uncertainty would, thus, typically require simulating multiphase flow and transport for a large number of permeability and release scenarios to collect statistics associated with remedial effectiveness, a computationally prohibitive process. The primary objective of this work is to develop and demonstrate a methodology that employs measured field data to produce equi-probable stochastic representations of a subsurface source zone that capture the spatial distribution and uncertainty associated with key features that control remediation performance (i.e., permeability and contamination mass). Here we employ probabilistic models known as discriminative random fields (DRFs) to synthesize stochastic realizations of initial mass distributions consistent with known, and typically limited, site characterization data. Using a limited number of full scale simulations as training data, a statistical model is developed for predicting the distribution of contaminant mass (e.g., DNAPL saturation and aqueous concentration) across a heterogeneous domain. Monte-Carlo sampling methods are then employed, in conjunction with the trained statistical model, to generate realizations conditioned on measured borehole data

  6. Molecular mechanisms of cell-cell spread of intracellular bacterial pathogens.

    Science.gov (United States)

    Ireton, Keith

    2013-07-17

    Several bacterial pathogens, including Listeria monocytogenes, Shigella flexneri and Rickettsia spp., have evolved mechanisms to actively spread within human tissues. Spreading is initiated by the pathogen-induced recruitment of host filamentous (F)-actin. F-actin forms a tail behind the microbe, propelling it through the cytoplasm. The motile pathogen then encounters the host plasma membrane, forming a bacterium-containing protrusion that is engulfed by an adjacent cell. Over the past two decades, much progress has been made in elucidating mechanisms of F-actin tail formation. Listeria and Shigella produce tails of branched actin filaments by subverting the host Arp2/3 complex. By contrast, Rickettsia forms tails with linear actin filaments through a bacterial mimic of eukaryotic formins. Compared with F-actin tail formation, mechanisms controlling bacterial protrusions are less well understood. However, recent findings have highlighted the importance of pathogen manipulation of host cell-cell junctions in spread. Listeria produces a soluble protein that enhances bacterial protrusions by perturbing tight junctions. Shigella protrusions are engulfed through a clathrin-mediated pathway at 'tricellular junctions'--specialized membrane regions at the intersection of three epithelial cells. This review summarizes key past findings in pathogen spread, and focuses on recent developments in actin-based motility and the formation and internalization of bacterial protrusions.

  7. BAR domain proteins regulate Rho GTPase signaling.

    Science.gov (United States)

    Aspenström, Pontus

    2014-01-01

    BAR proteins comprise a heterogeneous group of multi-domain proteins with diverse biological functions. The common denominator is the Bin-Amphiphysin-Rvs (BAR) domain that not only confers targeting to lipid bilayers, but also provides scaffolding to mold lipid membranes into concave or convex surfaces. This function of BAR proteins is an important determinant in the dynamic reconstruction of membrane vesicles, as well as of the plasma membrane. Several BAR proteins function as linkers between cytoskeletal regulation and membrane dynamics. These links are provided by direct interactions between BAR proteins and actin-nucleation-promoting factors of the Wiskott-Aldrich syndrome protein family and the Diaphanous-related formins. The Rho GTPases are key factors for orchestration of this intricate interplay. This review describes how BAR proteins regulate the activity of Rho GTPases, as well as how Rho GTPases regulate the function of BAR proteins. This mutual collaboration is a central factor in the regulation of vital cellular processes, such as cell migration, cytokinesis, intracellular transport, endocytosis, and exocytosis.

  8. Septation of infectious hyphae is critical for appressoria formation and virulence in the smut fungus Ustilago maydis.

    Directory of Open Access Journals (Sweden)

    Johannes Freitag

    2011-05-01

    Full Text Available Differentiation of hyphae into specialized infection structures, known as appressoria, is a common feature of plant pathogenic fungi that penetrate the plant cuticle. Appressorium formation in U. maydis is triggered by environmental signals but the molecular mechanism of this hyphal differentiation is largely unknown. Infectious hyphae grow on the leaf surface by inserting regularly spaced retraction septa at the distal end of the tip cell leaving empty sections of collapsed hyphae behind. Here we show that formation of retraction septa is critical for appressorium formation and virulence in U. maydis. We demonstrate that the diaphanous-related formin Drf1 is necessary for actomyosin ring formation during septation of infectious hyphae. Drf1 acts as an effector of a Cdc42 GTPase signaling module, which also consists of the Cdc42-specific guanine nucleotide exchange factor Don1 and the Ste20-like kinase Don3. Deletion of drf1, don1 or don3 abolished formation of retraction septa resulting in reduced virulence. Appressorium formation in these mutants was not completely blocked but infection structures were found only at the tip of short filaments indicating that retraction septa are necessary for appressorium formation in extended infectious hyphae. In addition, appressoria of drf1 mutants penetrated the plant tissue less frequently.

  9. Cdc42 controls primary mesenchyme cell morphogenesis in the sea urchin embryo.

    Science.gov (United States)

    Sepúlveda-Ramírez, Silvia P; Toledo-Jacobo, Leslie; Henson, John H; Shuster, Charles B

    2018-05-15

    In the sea urchin embryo, gastrulation is characterized by the ingression and directed cell migration of primary mesenchyme cells (PMCs), as well as the primary invagination and convergent extension of the endomesoderm. Like all cell shape changes, individual and collective cell motility is orchestrated by Rho family GTPases and their modulation of the actomyosin cytoskeleton. And while endomesoderm specification has been intensively studied in echinoids, much less is known about the proximate regulators driving cell motility. Toward these ends, we employed anti-sense morpholinos, mutant alleles and pharmacological inhibitors to assess the role of Cdc42 during sea urchin gastrulation. While inhibition of Cdc42 expression or activity had only mild effects on PMC ingression, PMC migration, alignment and skeletogenesis were disrupted in the absence of Cdc42, as well as elongation of the archenteron. PMC migration and patterning of the larval skeleton relies on the extension of filopodia, and Cdc42 was required for filopodia in vivo as well as in cultured PMCs. Lastly, filopodial extension required both Arp2/3 and formin actin-nucleating factors, supporting models of filopodial nucleation observed in other systems. Together, these results suggest that Cdc42 plays essential roles during PMC cell motility and organogenesis. Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.

  10. Dia-Interacting Protein (DIP) Imposes Migratory Plasticity in mDia2-Dependent Tumor Cells in Three-Dimensional Matrices

    Science.gov (United States)

    Wyse, Meghan M.; Lei, Jun; Nestor-Kalinoski, Andrea L.; Eisenmann, Kathryn M.

    2012-01-01

    Tumor cells rely upon membrane pliancy to escape primary lesions and invade secondary metastatic sites. This process relies upon localized assembly and disassembly cycles of F-actin that support and underlie the plasma membrane. Dynamic actin generates both spear-like and bleb structures respectively characterizing mesenchymal and amoeboid motility programs utilized by metastatic cells in three-dimensional matrices. The molecular mechanism and physiological trigger(s) driving membrane plasticity are poorly understood. mDia formins are F-actin assembly factors directing membrane pliancy in motile cells. mDia2 is functionally coupled with its binding partner DIP, regulating cortical actin and inducing membrane blebbing in amoeboid cells. Here we show that mDia2 and DIP co-tether to nascent blebs and this linkage is required for bleb formation. DIP controls mesenchymal/amoeboid cell interconvertability, while CXCL12 induces assembly of mDia2:DIP complexes to bleb cortices in 3D matrices. These results demonstrate how DIP-directed mDia2-dependent F-actin dynamics regulate morphological plasticity in motile cancer cells. PMID:23024796

  11. Dia-interacting protein (DIP imposes migratory plasticity in mDia2-dependent tumor cells in three-dimensional matrices.

    Directory of Open Access Journals (Sweden)

    Meghan M Wyse

    Full Text Available Tumor cells rely upon membrane pliancy to escape primary lesions and invade secondary metastatic sites. This process relies upon localized assembly and disassembly cycles of F-actin that support and underlie the plasma membrane. Dynamic actin generates both spear-like and bleb structures respectively characterizing mesenchymal and amoeboid motility programs utilized by metastatic cells in three-dimensional matrices. The molecular mechanism and physiological trigger(s driving membrane plasticity are poorly understood. mDia formins are F-actin assembly factors directing membrane pliancy in motile cells. mDia2 is functionally coupled with its binding partner DIP, regulating cortical actin and inducing membrane blebbing in amoeboid cells. Here we show that mDia2 and DIP co-tether to nascent blebs and this linkage is required for bleb formation. DIP controls mesenchymal/amoeboid cell interconvertability, while CXCL12 induces assembly of mDia2:DIP complexes to bleb cortices in 3D matrices. These results demonstrate how DIP-directed mDia2-dependent F-actin dynamics regulate morphological plasticity in motile cancer cells.

  12. Yeast G-proteins mediate directional sensing and polarization behaviors in response to changes in pheromone gradient direction

    Science.gov (United States)

    Moore, Travis I.; Tanaka, Hiromasa; Kim, Hyung Joon; Jeon, Noo Li; Yi, Tau-Mu

    2013-01-01

    Yeast cells polarize by projecting up mating pheromone gradients, a classic cell polarity behavior. However, these chemical gradients may shift direction. We examine how yeast cells sense and respond to a 180o switch in the direction of microfluidically generated pheromone gradients. We identify two behaviors: at low concentrations of α-factor, the initial projection grows by bending, whereas at high concentrations, cells form a second projection toward the new source. Mutations that increase heterotrimeric G-protein activity expand the bending-growth morphology to high concentrations; mutations that increase Cdc42 activity result in second projections at low concentrations. Gradient-sensing projection bending requires interaction between Gβγ and Cdc24, whereas gradient-nonsensing projection extension is stimulated by Bem1 and hyperactivated Cdc42. Of interest, a mutation in Gα affects both bending and extension. Finally, we find a genetic perturbation that exhibits both behaviors. Overexpression of the formin Bni1, a component of the polarisome, makes both bending-growth projections and second projections at low and high α-factor concentrations, suggesting a role for Bni1 downstream of the heterotrimeric G-protein and Cdc42 during gradient sensing and response. Thus we demonstrate that G-proteins modulate in a ligand-dependent manner two fundamental cell-polarity behaviors in response to gradient directional change. PMID:23242998

  13. Potential use of nuclear irradiation technology for the microbial industry in Malaysia

    International Nuclear Information System (INIS)

    Raja, P.; Isyanti, I.

    2002-01-01

    The potential application of irradiation is numerous. The process seems technically feasible for sterilization of medical devices, pharmaceutical products, foods, cosmetics, packaging materials, scientific laboratory materials like petri dishes, growth media, complying with phytosanitary requirements for fruit and vegetables etc. An awareness of biofertilizer or microbial product has established itself to be a significant part of food production and consumption internationally as well as in Malaysia. People are discovering organic agriculture can serve their need for safe quality food, environment conservation and social accountability. One of the major technical constraints of viable microbial or biofertilizer product production is contaminated free carrier materials. In order to develop the good microbial product, we are practicing the gamma irradiation on our carrier materials and successfully established the product. The different carrier materials were packed and used 25 kGy Cobalt-60 source (JS 8900, SINAGAMA, MINT) for sterilization and simultaneously the materials were sterilized in steam. The cultured Trichoderma virile and T harzianum were mixed with sterilized carrier and observed the colony formin. unit (CFU) on regular intervals. The results showed the efficient colony forming unit on both sterilization method of materials. However, the nuclear irradiation is more cost effective and time saving than steam method. (Author)

  14. Lumen Formation Is an Intrinsic Property of Isolated Human Pluripotent Stem Cells

    Directory of Open Access Journals (Sweden)

    Kenichiro Taniguchi

    2015-12-01

    Full Text Available We demonstrate that dissociated human pluripotent stem cells (PSCs are intrinsically programmed to form lumens. PSCs form two-cell cysts with a shared apical domain within 20 hr of plating; these cysts collapse to form monolayers after 5 days. Expression of pluripotency markers is maintained throughout this time. In two-cell cysts, an apical domain, marked by EZRIN and atypical PKCζ, is surrounded by apically targeted organelles (early endosomes and Golgi. Molecularly, actin polymerization, regulated by ARP2/3 and mammalian diaphanous-related formin 1 (MDIA, promotes lumen formation, whereas actin contraction, mediated by MYOSIN-II, inhibits this process. Finally, we show that lumenal shape can be manipulated in bioengineered micro-wells. Since lumen formation is an indispensable step in early mammalian development, this system can provide a powerful model for investigation of this process in a controlled environment. Overall, our data establish that lumenogenesis is a fundamental cell biological property of human PSCs.

  15. Paleocene to Middle Miocene planktic foraminifera of the southwestern Salisbury Embayment, Virginia and Maryland: biostratigraphy, allostratigraphy, and sequence stratigraphy

    Science.gov (United States)

    Poag, C.W.; Commeau, J.A.

    1995-01-01

    The Paleocene to Middle Miocene sedimentary fill of the southwestern Salisbury Embayment contains a fragmental depositional record, interrupted by numerous local diastems and regional unconformities. Using planktic foraminiferal biostratigraphy, 15 unconformity-bounded depositional units have been identified, assigned to six formations and seven alloformations previously recognized in the embayment. The units correlate with second- and third-order sequences of the Exxon sequence stratigraphy model, and include transgressive and highstand systems tracts. Alloformation, formation, and sequence boundaries are marked by abrupt, scoured, burrowed, erosional surfaces, which display lag deposits, biostratigraphic gaps, and intense reworking of microfossils above and below the boundaries.Paleocene deposits represent the upper parts of upper Pleocene Biochronozones P4 and P5, and rest uncomformably  on Cretaceous sedimentary beds of various ages (Maastrichtian to Albian). Lower Eocene deposits represent parts of Biochronozones P6 and P9. Middle Eocene strata represent mainly parts of Biochronozones P11, P12, and P14. Upper Eocene sediments include parts of Biochronozones P15, P16, and P17. Oligocene deposits encompass parts of Biochronozones. N4b to N7 undifferentiated, P21a, and, perhaps, N4a. Lower Miocene deposits encompass parts of Biochronozones N4b to N7 undifferentiated. Middle Miocene strata represent mainly parts of Biochronorones N8, N9, and N10.Nine plates of scanning electron micrographs illustrate the principal planktic foraminifera used to establish the biostratigraphic framework. Two new informal formine of Praeterenuitella praegemma Li, 1987, are introduced.

  16. Variations Method to Solve Terminal Problems for the Second Order Systems of Canonical Form with State Constraints

    Directory of Open Access Journals (Sweden)

    T. S. Kasatkina

    2015-01-01

    Full Text Available Terminal control problem with fixed finite time for the second order affine systems with state constraints is considered. A solution of such terminal problem is suggested for the systems with scalar control of regular canonical form.In this article it is shown that the initial terminal problem is equivalent to the problem of auxiliary function search. This function should satisfy some conditions. Such function design consists of two stages. The first stage includes search of function which corresponds the solution of the terminal control problem without state constraints. This function is designed as polynom of the fifth power which depends on time variable. Coefficients of the polynom are defined by boundary conditions. The second stage includes modification of designed function if corresponding to that function trajectory is not satisfied constraints. Modification process is realized by adding to the current function supplementary polynom. Influence of that polynom handles by variation of a parameter value. Modification process can include a few iterations. After process termination continuous control is found. This control is the solution of the initial terminal prUsing presented scheme the terminal control problem for system, which describes oscillations of the mathematical pendulum, is solved. This approach can be used for the solution of terminal control problems with state constraints for affine systems with multi-dimensional control.

  17. Radical Roles for RAGE in the Pathogenesis of Oxidative Stress in Cardiovascular Diseases and Beyond

    Directory of Open Access Journals (Sweden)

    Radha Ananthakrishnan

    2013-10-01

    Full Text Available Oxidative stress is a central mechanism by which the receptor for advanced glycation endproducts (RAGE mediates its pathological effects. Multiple experimental inquiries in RAGE-expressing cultured cells have demonstrated that ligand-RAGE interaction mediates generation of reactive oxygen species (ROS and consequent downstream signal transduction and regulation of gene expression. The primary mechanism by which RAGE generates oxidative stress is via activation of NADPH oxidase; amplification mechanisms in the mitochondria may further drive ROS production. Recent studies indicating that the cytoplasmic domain of RAGE binds to the formin mDia1 provide further support for the critical roles of this pathway in oxidative stress; mDia1 was required for activation of rac1 and NADPH oxidase in primary murine aortic smooth muscle cells treated with RAGE ligand S100B. In vivo, in multiple distinct disease models in animals, RAGE action generates oxidative stress and modulates cellular/tissue fate in range of disorders, such as in myocardial ischemia, atherosclerosis, and aneurysm formation. Blockade or genetic deletion of RAGE was shown to be protective in these settings. Indeed, beyond cardiovascular disease, evidence is accruing in human subjects linking levels of RAGE ligands and soluble RAGE to oxidative stress in disorders such as doxorubicin toxicity, acetaminophen toxicity, neurodegeneration, hyperlipidemia, diabetes, preeclampsia, rheumatoid arthritis and pulmonary fibrosis. Blockade of RAGE signal transduction may be a key strategy for the prevention of the deleterious consequences of oxidative stress, particularly in chronic disease.

  18. Code of practice for radiation protection in nuclear medicine

    International Nuclear Information System (INIS)

    Hamed, M. I.

    2010-05-01

    In aim of this study was to develop a draft for a new code practice for radiation protection in nuclear medicine that meets the current relevant international recommendation. The draft includes the following main fields: methods of radiation protection for workers, patients and public. Also, the principles of safe design of nuclear medicine departments, quality assurance program, proper manipulation of radiation sources including radioactive waste and emergency preparedness and response. The practical part of this study includes inspections of three nuclear medicine departments available in Sudan so as to assess the degree of compliance of those departments with what is stated in this code. The inspection missions have been conducted using a checklist that addresses all items that may affect radiation raincoat issues in addition to per formin area radiation monitoring around the installation of the radioactive sources. The results of this revealed that most of the departments do not have effective radiation protection program which in turn could lead to unnecessary exposure to patients, public and workers. Finally, some recommendations are given that - if implemented - could improve the status of radiation protection in nuclear medicine department. (Author)

  19. Integrin-based diffusion barrier separates membrane domains enabling the formation of microbiostatic frustrated phagosomes

    Science.gov (United States)

    Maxson, Michelle E; Naj, Xenia; O'Meara, Teresa R; Plumb, Jonathan D; Cowen, Leah E

    2018-01-01

    Candida albicans hyphae can reach enormous lengths, precluding their internalization by phagocytes. Nevertheless, macrophages engulf a portion of the hypha, generating incompletely sealed tubular phagosomes. These frustrated phagosomes are stabilized by a thick cuff of F-actin that polymerizes in response to non-canonical activation of integrins by fungal glycan. Despite their continuity, the surface and invaginating phagosomal membranes retain a strikingly distinct lipid composition. PtdIns(4,5)P2 is present at the plasmalemma but is not detectable in the phagosomal membrane, while PtdIns(3)P and PtdIns(3,4,5)P3 co-exist in the phagosomes yet are absent from the surface membrane. Moreover, endo-lysosomal proteins are present only in the phagosomal membrane. Fluorescence recovery after photobleaching revealed the presence of a diffusion barrier that maintains the identity of the open tubular phagosome separate from the plasmalemma. Formation of this barrier depends on Syk, Pyk2/Fak and formin-dependent actin assembly. Antimicrobial mechanisms can thereby be deployed, limiting the growth of the hyphae. PMID:29553370

  20. Septation of infectious hyphae is critical for appressoria formation and virulence in the smut fungus Ustilago maydis.

    Science.gov (United States)

    Freitag, Johannes; Lanver, Daniel; Böhmer, Christian; Schink, Kay Oliver; Bölker, Michael; Sandrock, Björn

    2011-05-01

    Differentiation of hyphae into specialized infection structures, known as appressoria, is a common feature of plant pathogenic fungi that penetrate the plant cuticle. Appressorium formation in U. maydis is triggered by environmental signals but the molecular mechanism of this hyphal differentiation is largely unknown. Infectious hyphae grow on the leaf surface by inserting regularly spaced retraction septa at the distal end of the tip cell leaving empty sections of collapsed hyphae behind. Here we show that formation of retraction septa is critical for appressorium formation and virulence in U. maydis. We demonstrate that the diaphanous-related formin Drf1 is necessary for actomyosin ring formation during septation of infectious hyphae. Drf1 acts as an effector of a Cdc42 GTPase signaling module, which also consists of the Cdc42-specific guanine nucleotide exchange factor Don1 and the Ste20-like kinase Don3. Deletion of drf1, don1 or don3 abolished formation of retraction septa resulting in reduced virulence. Appressorium formation in these mutants was not completely blocked but infection structures were found only at the tip of short filaments indicating that retraction septa are necessary for appressorium formation in extended infectious hyphae. In addition, appressoria of drf1 mutants penetrated the plant tissue less frequently.

  1. El perfil de l’emprenedor social de l’estudiantat dels Graus d’Educació Social, Pedagogia i Treball Social a la Universitat de Barcelona

    Directory of Open Access Journals (Sweden)

    Olga Cabrera-Santacana

    2014-01-01

    Full Text Available En el context de crisi actual, cada vegada més, es posa més atenció a un tipus d'economia més sostenible i amb valors socials, i la Universitat ha de reconèixer la seva responsabilitat en aquests nous reptes. Durant els darrers cursos acadèmics compresos entre 2010 i 2012, i amb el suport econòmic del projecte REDICE 2010, s'ha analitzat i identificat les necessitats formatives del perfil emprenedor social del nostre alumnat de la Facultat de Pedagogia, s'ha pogut anar introduint el concepte d'Emprenedoria social dins dels Graus d'Educació Social, Pedagogia i Treball Social a través de bones pràctiques que formin perfils universitaris creatius i innovadors per a l’emprenedoria social. Com a resultat d'aquesta anàlisi es mostra un interès per l'emprenedoria social i per unes necessitats acadèmiques per millorar la nostra activitat docent, buscant bones pràctiques identificades en aquesta línia per millorar la formació i docència en relació l'emprenedoria social.

  2. Cytoskeleton in Mast Cell Signaling

    Science.gov (United States)

    Dráber, Pavel; Sulimenko, Vadym; Dráberová, Eduarda

    2012-01-01

    Mast cell activation mediated by the high affinity receptor for IgE (FcεRI) is a key event in allergic response and inflammation. Other receptors on mast cells, as c-Kit for stem cell factor and G protein-coupled receptors (GPCRs) synergistically enhance the FcεRI-mediated release of inflammatory mediators. Activation of various signaling pathways in mast cells results in changes in cell morphology, adhesion to substrate, exocytosis, and migration. Reorganization of cytoskeleton is pivotal in all these processes. Cytoskeletal proteins also play an important role in initial stages of FcεRI and other surface receptors induced triggering. Highly dynamic microtubules formed by αβ-tubulin dimers as well as microfilaments build up from polymerized actin are affected in activated cells by kinases/phosphatases, Rho GTPases and changes in concentration of cytosolic Ca2+. Also important are nucleation proteins; the γ-tubulin complexes in case of microtubules or Arp 2/3 complex with its nucleation promoting factors and formins in case of microfilaments. The dynamic nature of microtubules and microfilaments in activated cells depends on many associated/regulatory proteins. Changes in rigidity of activated mast cells reflect changes in intermediate filaments build up from vimentin. This review offers a critical appraisal of current knowledge on the role of cytoskeleton in mast cells signaling. PMID:22654883

  3. Identification of Diabetic Retinopathy Genes through a Genome-Wide Association Study among Mexican-Americans from Starr County, Texas

    Directory of Open Access Journals (Sweden)

    Yi-Ping Fu

    2010-01-01

    Full Text Available To identify genetic loci for severe diabetic retinopathy, 286 Mexican-Americans with type 2 diabetes from Starr County, Texas, completed physical examinations including fundus photography for diabetic retinopathy grading. Individuals with moderate-to-severe non-proliferative and proliferative diabetic retinopathy were defined as cases. Direct genotyping was performed using the Affymetrix GeneChip Human Mapping 100 K Set, and SNPs passing quality control criteria were used to impute markers available in HapMap Phase III Mexican population (MXL in Los Angeles, California. Two directly genotyped markers were associated with severe diabetic retinopathy at a P-value less than .0001: SNP rs2300782 (P=6.04×10−5 mapped to an intron region of CAMK4 (calcium/calmodulin-dependent protein kinase IV on chromosome 5, and SNP rs10519765 (P=6.21×10−5 on chromosomal 15q13 in the FMN1 (formin 1 gene. Using well-imputed markers based on the HapMap III Mexican population, we identified an additional 32 SNPs located in 11 chromosomal regions with nominal association with severe diabetic retinopathy at P-value less than .0001. None of these markers were located in traditional candidate genes for diabetic retinopathy or diabetes itself. However, these signals implicate genes involved in inflammation, oxidative stress and cell adhesion for the development and progression of diabetic retinopathy.

  4. Two Functionally Distinct Sources of Actin Monomers Supply the Leading Edge of Lamellipodia

    Science.gov (United States)

    Vitriol, Eric A.; McMillen, Laura M.; Kapustina, Maryna; Gomez, Shawn M.; Vavylonis, Dimitrios; Zheng, James Q.

    2015-01-01

    Summary Lamellipodia, the sheet-like protrusions of motile cells, consist of networks of actin filaments (F-actin) regulated by the ordered assembly from and disassembly into actin monomers (G-actin). Traditionally, G-actin is thought to exist as a homogeneous pool. Here, we show that there are two functionally and molecularly distinct sources of G-actin that supply lamellipodial actin networks. G-actin originating from the cytosolic pool requires the monomer binding protein thymosin β4 (Tβ4) for optimal leading edge localization, is targeted to formins, and is responsible for creating an elevated G/F-actin ratio that promotes membrane protrusion. The second source of G-actin comes from recycled lamellipodia F-actin. Recycling occurs independently of Tβ4 and appears to regulate lamellipodia homeostasis. Tβ4-bound G-actin specifically localizes to the leading edge because it doesn’t interact with Arp2/3-mediated polymerization sites found throughout the lamellipodia. These findings demonstrate that actin networks can be constructed from multiple sources of monomers with discrete spatiotemporal functions. PMID:25865895

  5. Sequential actin-based pushing forces drive meiosis I chromosome migration and symmetry breaking in oocytes

    Science.gov (United States)

    Yi, Kexi; Rubinstein, Boris; Unruh, Jay R.; Guo, Fengli; Slaughter, Brian D.

    2013-01-01

    Polar body extrusion during oocyte maturation is critically dependent on asymmetric positioning of the meiotic spindle, which is established through migration of the meiosis I (MI) spindle/chromosomes from the oocyte interior to a subcortical location. In this study, we show that MI chromosome migration is biphasic and driven by consecutive actin-based pushing forces regulated by two actin nucleators, Fmn2, a formin family protein, and the Arp2/3 complex. Fmn2 was recruited to endoplasmic reticulum structures surrounding the MI spindle, where it nucleated actin filaments to initiate an initially slow and poorly directed motion of the spindle away from the cell center. A fast and highly directed second migration phase was driven by actin-mediated cytoplasmic streaming and occurred as the chromosomes reach a sufficient proximity to the cortex to activate the Arp2/3 complex. We propose that decisive symmetry breaking in mouse oocytes results from Fmn2-mediated perturbation of spindle position and the positive feedback loop between chromosome signal-induced Arp2/3 activation and Arp2/3-orchestrated cytoplasmic streaming that transports the chromosomes. PMID:23439682

  6. The radiomodifying efficacy of beta carotene rich plant extracts on neuroethology of Swiss albino mice: perception, perspectives and risk assessment

    International Nuclear Information System (INIS)

    Bhatia, A.L.

    2003-01-01

    Full text: High utilization of O2 and rather poorly developed antioxidative defence mechanism makes the brain highly susceptible to oxidative damage. High enrichment with PUFA also renders it susceptible to radiation damage by free radicals. The pure form of beta carotene has proved quite effective against radiation but only at optimum dose level when tested for survivability and lipid peroxidation, protein, cholesterol, DNA content of brain. This induced us to extend our investigation on plants, Amaranthus and Spinach enriched with beta carotene, which could be recommended in the nutritional dietary course without causing psychological stress of availability and affordability unlike of tablets of medicines. Both Amaranthus paniculatus and Spinacea oleracea, commonly occurring weeds have good nutritive values due to their carotenoid, vitamin C, folate, folic acid contents; additionally Amaranthus with high level of lysine and methionine. Swiss albino male mice of 6-8 week(22±3 gm)selected from an inbred colony were administered with alcoholic extract at a dose of 600-mg/kg-body weight/day dissolved in distilled water with and without prior to irradiation (5 Gy of gamma radiation). The animals were studied on 1, 3, 7, 15, and 30 days after radiation exposure. On the basis of LD50/30 values the DRFs were computed as 1.43(AE) and 1.39(S.E). The plant extracts improved learning performance in mice in with and without rradiation. Male mice showed better learning performance as compared to females in all the groups. The brain showed that the radiation induced depletion of protein, glutathione and cholesterol and histopathology was significantly compensated/defied and was brought to near-normal level by the 15 days oral administration of crude extract of the plants. Radiation induced augmentation in glycogen, cholesterol and lipid peroxidation products were significantly checked. The protection appears to be afforded by combined or synergistic effects of plants leaves

  7. Validation and uncertainty quantification of detector response functions for a 1″×2″ NaI collimated detector intended for inverse radioisotope source mapping applications

    Science.gov (United States)

    Nelson, N.; Azmy, Y.; Gardner, R. P.; Mattingly, J.; Smith, R.; Worrall, L. G.; Dewji, S.

    2017-11-01

    Detector response functions (DRFs) are often used for inverse analysis. We compute the DRF of a sodium iodide (NaI) nuclear material holdup field detector using the code named g03 developed by the Center for Engineering Applications of Radioisotopes (CEAR) at NC State University. Three measurement campaigns were performed in order to validate the DRF's constructed by g03: on-axis detection of calibration sources, off-axis measurements of a highly enriched uranium (HEU) disk, and on-axis measurements of the HEU disk with steel plates inserted between the source and the detector to provide attenuation. Furthermore, this work quantifies the uncertainty of the Monte Carlo simulations used in and with g03, as well as the uncertainties associated with each semi-empirical model employed in the full DRF representation. Overall, for the calibration source measurements, the response computed by the DRF for the prediction of the full-energy peak region of responses was good, i.e. within two standard deviations of the experimental response. In contrast, the DRF tended to overestimate the Compton continuum by about 45-65% due to inadequate tuning of the electron range multiplier fit variable that empirically represents physics associated with electron transport that is not modeled explicitly in g03. For the HEU disk measurements, computed DRF responses tended to significantly underestimate (more than 20%) the secondary full-energy peaks (any peak of lower energy than the highest-energy peak computed) due to scattering in the detector collimator and aluminum can, which is not included in the g03 model. We ran a sufficiently large number of histories to ensure for all of the Monte Carlo simulations that the statistical uncertainties were lower than their experimental counterpart's Poisson uncertainties. The uncertainties associated with least-squares fits to the experimental data tended to have parameter relative standard deviations lower than the peak channel relative standard

  8. Design of a head phantom produced on a 3D rapid prototyping printer and comparison with a RANDO and 3M lucite head phantom in eye dosimetry applications.

    Science.gov (United States)

    Homolka, Peter; Figl, Michael; Wartak, Andreas; Glanzer, Mathias; Dünkelmeyer, Martina; Hojreh, Azadeh; Hummel, Johann

    2017-04-21

    An anthropomorphic head phantom including eye inserts allowing placement of TLDs 3 mm below the cornea has been produced on a 3D printer using a photo-cured acrylic resin to best allow tissue equivalence. Thus H p (3) can be determined in radiological and interventional photon radiation fields. Eye doses and doses to the forehead have been compared to an Alderson RANDO head and a 3M Lucite skull phantom in terms of surface dose per incident air kerma for frontal irradiation since the commercial phantoms do not allow placement of TLDs 3 mm below the corneal surface. A comparison of dose reduction factors (DRFs) of a common lead glasses model has also been performed. Eye dose per incident air kerma were comparable between all three phantoms (printed phantom: 1.40, standard error (SE) 0.04; RANDO: 1.36, SE 0.03; 3M: 1.37, SE 0.03). Doses to the forehead were identical to eye surface doses for the printed phantom and the RANDO head (ratio 1.00 SE 0.04, and 0.99 SE 0.03, respectively). In the 3M Lucite skull phantom dose on the forehead was 15% lower than dose to the eyes attributable to phantom properties. DRF of a sport frame style leaded glasses model with 0.75 mm lead equivalence measured were 6.8 SE 0.5, 9.3 SE 0.4 and 10.5 SE 0.5 for the RANDO head, the printed phantom, and the 3M Lucite head phantom, respectively, for frontal irradiation. A comparison of doses measured in 3 mm depth and on the surface of the eyes in the printed phantom revealed no difference larger than standard errors from TLD dosimetry. 3D printing offers an interesting opportunity for phantom design with increasing potential as printers allowing combinations of tissue substitutes will become available. Variations between phantoms may provide a useful indication of uncertainty budgets when using phantom measurements to estimate individual personnel doses.

  9. Molecular and Cellular Mechanisms of Shigella flexneri Dissemination.

    Science.gov (United States)

    Agaisse, Hervé

    2016-01-01

    The intracellular pathogen Shigella flexneri is the causative agent of bacillary dysentery in humans. The disease is characterized by bacterial invasion of intestinal cells, dissemination within the colonic epithelium through direct spread from cell to cell, and massive inflammation of the intestinal mucosa. Here, we review the mechanisms supporting S. flexneri dissemination. The dissemination process primarily relies on actin assembly at the bacterial pole, which propels the pathogen throughout the cytosol of primary infected cells. Polar actin assembly is supported by polar expression of the bacterial autotransporter family member IcsA, which recruits the N-WASP/ARP2/3 actin assembly machinery. As motile bacteria encounter cell-cell contacts, they form plasma membrane protrusions that project into adjacent cells. In addition to the ARP2/3-dependent actin assembly machinery, protrusion formation relies on formins and myosins. The resolution of protrusions into vacuoles occurs through the collapse of the protrusion neck, leading to the formation of an intermediate membrane-bound compartment termed vacuole-like protrusions (VLPs). VLP formation requires tyrosine kinase and phosphoinositide signaling in protrusions, which relies on the integrity of the bacterial type 3 secretion system (T3SS). The T3SS is also required for escaping double membrane vacuoles through the activity of the T3SS translocases IpaB and IpaC, and the effector proteins VirA and IcsB. Numerous factors supporting envelope biogenesis contribute to IcsA exposure and maintenance at the bacterial pole, including LPS synthesis, membrane proteases, and periplasmic chaperones. Although less characterized, the assembly and function of the T3SS in the context of bacterial dissemination also relies on factors supporting envelope biogenesis. Finally, the dissemination process requires the adaptation of the pathogen to various cellular compartments through transcriptional and post-transcriptional mechanisms.

  10. Binding of Cu(II) ions to peptides studied by fluorescence spectroscopy and isothermal titration calorimetry

    Science.gov (United States)

    Makowska, Joanna; Żamojć, Krzysztof; Wyrzykowski, Dariusz; Uber, Dorota; Wierzbicka, Małgorzata; Wiczk, Wiesław; Chmurzyński, Lech

    2016-01-01

    Steady-state and time-resolved fluorescence quenching measurements supported by Isothermal Titration Calorimetry (ITC) were used to study the interactions of Cu2 + with four peptides. Two of them were taken from the N-terminal part of the FBP28 protein (formin binding protein) WW domain: Tyr-Lys-Thr-Ala-Asp-Gly-Lys-Thr-Tyr-NH2 (D9) and its mutant Tyr-Lys-Thr-Ala-Asn-Gly-Lys-Thr-Tyr-NH2 (D9_M) as well as two mutated peptides from the B3 domain of the immunoglobulin binding protein G derived from Streptococcus: Asp-Val-Ala-Thr-Tyr-Thr-NH2 (J1) and Glu-Val-Ala-Thr-Tyr-Thr-NH2 (J2). The measurements were carried out at 298.15 K in 20 mM 2-(N-morpholino)ethanesulfonic acid (MES) buffer solution with a pH of 6. The fluorescence of all peptides was quenched by Cu2 + ions. The stoichiometry, conditional stability constants and thermodynamic parameters for the interactions of the Cu2 + ions with D9 and D9_M were determined from the calorimetric data. The values of the conditional stability constants were additionally determined from fluorescence quenching measurements and compared with those obtained from calorimetric studies. There was a good correlation between data obtained from the two techniques. On the other hand, the studies revealed that J1 and J2 do not exhibit an affinity towards metal ions. The obtained results prove that fluorescence quenching experiments may be successfully used in order to determine stability constants of complexes with fluorescent ligands. Finally, based on the obtained results, the coordinating properties of the peptides towards the Cu2 + ions are discussed.

  11. Carbonated water (CW) process waste reuse for ammonium-uranyl-carbonate (AUC) production and its gains on the environmental, economic and social aspects

    International Nuclear Information System (INIS)

    Carnaval, Joao Paulo R.; Santos, Rafael D. dos; Barbosa, Rodrigo A.; Lauer, Sergio

    2013-01-01

    In the INB nuclear fuel cycle, the pellets production is based on UO 2 powder made by AUC (Ammonium-Uranyl-Carbonate) route. AUC formation occurs by fluidising of UF 6 , NH 3 and CO 2 in a vase containing usually pure water, and this exothermal reaction has AUC as direct product. The mass formed is filtered, washed with CW, washed again with methano solution, dried with air and conducted to the fluidized bed furnace, to be converted to UO 2 powder. At this point, the dried AUC decompounds to UO 3 , NH 3 and C0 2 , these 2 gases are absorbed at the gases washer, formin go the carbonated water (CW), whit is basically a (NH 4 ) 2 CO 3 solution. The UO 2+x is reduced and stabilized to UO 2 powder, which is conducted to pellets production. During the process, a considerable amount of this aqueous waste is generated and goes for effluent treatment. After that, the solution is sent for spray-dryer for power formation, and stock. This treatment demands equipment, energy and time, representing considerable costs of the company beyond the human risks involved on the drying step. The purpose of this work is to present a study of the carbonated water use as substitute of pure water in the AUC formation step. At this point, tests were made varying the CW loads for the AUC precipitation, and the control was made by the UO 2 powder properties. The carbonated water used for AUC precipitation has been tested at several levels and the results has demonstrated full viability to become a definitive process step (INB, Resende site). It has been demonstrated the great resources economy caused by the waste reuse and the guarantee product quality. This represents such an environmental gain and also economic and social aspects got improved. (author)

  12. Proline: the distribution, frequency, positioning, and common functional roles of proline and polyproline sequences in the human proteome.

    Directory of Open Access Journals (Sweden)

    Alexander A Morgan

    Full Text Available Proline is an anomalous amino acid. Its nitrogen atom is covalently locked within a ring, thus it is the only proteinogenic amino acid with a constrained phi angle. Sequences of three consecutive prolines can fold into polyproline helices, structures that join alpha helices and beta pleats as architectural motifs in protein configuration. Triproline helices are participants in protein-protein signaling interactions. Longer spans of repeat prolines also occur, containing as many as 27 consecutive proline residues. Little is known about the frequency, positioning, and functional significance of these proline sequences. Therefore we have undertaken a systematic bioinformatics study of proline residues in proteins. We analyzed the distribution and frequency of 687,434 proline residues among 18,666 human proteins, identifying single residues, dimers, trimers, and longer repeats. Proline accounts for 6.3% of the 10,882,808 protein amino acids. Of all proline residues, 4.4% are in trimers or longer spans. We detected patterns that influence function based on proline location, spacing, and concentration. We propose a classification based on proline-rich, polyproline-rich, and proline-poor status. Whereas singlet proline residues are often found in proteins that display recurring architectural patterns, trimers or longer proline sequences tend be associated with the absence of repetitive structural motifs. Spans of 6 or more are associated with DNA/RNA processing, actin, and developmental processes. We also suggest a role for proline in Kruppel-type zinc finger protein control of DNA expression, and in the nucleation and translocation of actin by the formin complex.

  13. Políticas de formación de profesores de educación secundaria en España y en Brasil : estudio comparado sobre tendencias de mercado en el contexto institucional

    Directory of Open Access Journals (Sweden)

    Rose Meri Trojan

    2011-10-01

    Full Text Available Los estudios comparativos han sido el objetivo de los gobiernos y de las organizaciones multilaterales, adquiriendo cada vez más relieve en el actual proceso de globalización. En la evaluación de las políticas, especialmente las educativas, la comparación se utiliza para delimitar los parámetros de calidad y los modelos de eficiencia. Este artículo tiene como objetivo presentar un estudio de las políticas en materia de formación y desarrollo profesional para profesores de enseñanza secundaria de España y Brasil, con las categorías de análisis de la descentralización, la financiación, la evaluación y cambios del perfil del profesor, bajo la influencia de los procesos de globalización y reforma del Estado en la educación, con el fin de identificar las tendencias en curso.Comparative studies have been widely used by governments and multilateral organizations and have been getting increasingly attention in the current process of globalization.In the evaluation of policies, especially the educational ones, the comparison is used to demarcate the parameters ofquality and models of efficiency. This article aims to present a study of professional formation policies and professional development of secondary teachers from Spain and Brazil.The categories of analysis used in this study are decentralization, financing, evaluation and the changes in the teachers’profiles as a result of the influence of the globalization processes and there formin education lead by the State, in order to identify ongoing trends.

  14. Mobilization of HIV Spread by Diaphanous 2 Dependent Filopodia in Infected Dendritic Cells

    Science.gov (United States)

    Aggarwal, Anupriya; Iemma, Tina L.; Shih, Ivy; Newsome, Timothy P.; McAllery, Samantha; Cunningham, Anthony L.; Turville, Stuart G.

    2012-01-01

    Paramount to the success of persistent viral infection is the ability of viruses to navigate hostile environments en route to future targets. In response to such obstacles, many viruses have developed the ability of establishing actin rich-membrane bridges to aid in future infections. Herein through dynamic imaging of HIV infected dendritic cells, we have observed how viral high-jacking of the actin/membrane network facilitates one of the most efficient forms of HIV spread. Within infected DC, viral egress is coupled to viral filopodia formation, with more than 90% of filopodia bearing immature HIV on their tips at extensions of 10 to 20 µm. Live imaging showed HIV filopodia routinely pivoting at their base, and projecting HIV virions at µm.sec−1 along repetitive arc trajectories. HIV filopodial dynamics lead to up to 800 DC to CD4 T cell contacts per hour, with selection of T cells culminating in multiple filopodia tethering and converging to envelope the CD4 T-cell membrane with budding HIV particles. Long viral filopodial formation was dependent on the formin diaphanous 2 (Diaph2), and not a dominant Arp2/3 filopodial pathway often associated with pathogenic actin polymerization. Manipulation of HIV Nef reduced HIV transfer 25-fold by reducing viral filopodia frequency, supporting the potency of DC HIV transfer was dependent on viral filopodia abundance. Thus our observations show HIV corrupts DC to CD4 T cell interactions by physically embedding at the leading edge contacts of long DC filopodial networks. PMID:22685410

  15. High-throughput transcriptome analysis of barley (Hordeum vulgare) exposed to excessive boron.

    Science.gov (United States)

    Tombuloglu, Guzin; Tombuloglu, Huseyin; Sakcali, M Serdal; Unver, Turgay

    2015-02-15

    Boron (B) is an essential micronutrient for optimum plant growth. However, above certain threshold B is toxic and causes yield loss in agricultural lands. While a number of studies were conducted to understand B tolerance mechanism, a transcriptome-wide approach for B tolerant barley is performed here for the first time. A high-throughput RNA-Seq (cDNA) sequencing technology (Illumina) was used with barley (Hordeum vulgare), yielding 208 million clean reads. In total, 256,874 unigenes were generated and assigned to known peptide databases: Gene Ontology (GO) (99,043), Swiss-Prot (38,266), Clusters of Orthologous Groups (COG) (26,250), and the Kyoto Encyclopedia of Genes and Genomes (KEGG) (36,860), as determined by BLASTx search. According to the digital gene expression (DGE) analyses, 16% and 17% of the transcripts were found to be differentially regulated in root and leaf tissues, respectively. Most of them were involved in cell wall, stress response, membrane, protein kinase and transporter mechanisms. Some of the genes detected as highly expressed in root tissue are phospholipases, predicted divalent heavy-metal cation transporters, formin-like proteins and calmodulin/Ca(2+)-binding proteins. In addition, chitin-binding lectin precursor, ubiquitin carboxyl-terminal hydrolase, and serine/threonine-protein kinase AFC2 genes were indicated to be highly regulated in leaf tissue upon excess B treatment. Some pathways, such as the Ca(2+)-calmodulin system, are activated in response to B toxicity. The differential regulation of 10 transcripts was confirmed by qRT-PCR, revealing the tissue-specific responses against B toxicity and their putative function in B-tolerance mechanisms. Copyright © 2014. Published by Elsevier B.V.

  16. Binding of Cu(II) ions to peptides studied by fluorescence spectroscopy and isothermal titration calorimetry.

    Science.gov (United States)

    Makowska, Joanna; Żamojć, Krzysztof; Wyrzykowski, Dariusz; Uber, Dorota; Wierzbicka, Małgorzata; Wiczk, Wiesław; Chmurzyński, Lech

    2016-01-15

    Steady-state and time-resolved fluorescence quenching measurements supported by Isothermal Titration Calorimetry (ITC) were used to study the interactions of Cu(2+) with four peptides. Two of them were taken from the N-terminal part of the FBP28 protein (formin binding protein) WW domain: Tyr-Lys-Thr-Ala-Asp-Gly-Lys-Thr-Tyr-NH2 (D9) and its mutant Tyr-Lys-Thr-Ala-Asn-Gly-Lys-Thr-Tyr-NH2 (D9_M) as well as two mutated peptides from the B3 domain of the immunoglobulin binding protein G derived from Streptococcus: Asp-Val-Ala-Thr-Tyr-Thr-NH2 (J1) and Glu-Val-Ala-Thr-Tyr-Thr-NH2 (J2). The measurements were carried out at 298.15K in 20mM 2-(N-morpholino)ethanesulfonic acid (MES) buffer solution with a pH of 6. The fluorescence of all peptides was quenched by Cu(2+) ions. The stoichiometry, conditional stability constants and thermodynamic parameters for the interactions of the Cu(2+) ions with D9 and D9_M were determined from the calorimetric data. The values of the conditional stability constants were additionally determined from fluorescence quenching measurements and compared with those obtained from calorimetric studies. There was a good correlation between data obtained from the two techniques. On the other hand, the studies revealed that J1 and J2 do not exhibit an affinity towards metal ions. The obtained results prove that fluorescence quenching experiments may be successfully used in order to determine stability constants of complexes with fluorescent ligands. Finally, based on the obtained results, the coordinating properties of the peptides towards the Cu(2+) ions are discussed. Copyright © 2015 Elsevier B.V. All rights reserved.

  17. Loss of γ-cytoplasmic actin triggers myofibroblast transition of human epithelial cells.

    Science.gov (United States)

    Lechuga, Susana; Baranwal, Somesh; Li, Chao; Naydenov, Nayden G; Kuemmerle, John F; Dugina, Vera; Chaponnier, Christine; Ivanov, Andrei I

    2014-10-15

    Transdifferentiation of epithelial cells into mesenchymal cells and myofibroblasts plays an important role in tumor progression and tissue fibrosis. Such epithelial plasticity is accompanied by dramatic reorganizations of the actin cytoskeleton, although mechanisms underlying cytoskeletal effects on epithelial transdifferentiation remain poorly understood. In the present study, we observed that selective siRNA-mediated knockdown of γ-cytoplasmic actin (γ-CYA), but not β-cytoplasmic actin, induced epithelial-to-myofibroblast transition (EMyT) of different epithelial cells. The EMyT manifested by increased expression of α-smooth muscle actin and other contractile proteins, along with inhibition of genes responsible for cell proliferation. Induction of EMyT in γ-CYA-depleted cells depended on activation of serum response factor and its cofactors, myocardial-related transcriptional factors A and B. Loss of γ-CYA stimulated formin-mediated actin polymerization and activation of Rho GTPase, which appear to be essential for EMyT induction. Our findings demonstrate a previously unanticipated, unique role of γ-CYA in regulating epithelial phenotype and suppression of EMyT that may be essential for cell differentiation and tissue fibrosis. © 2014 Lechuga, Baranwal, et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

  18. Comparative analysis of human conjunctival and corneal epithelial gene expression with oligonucleotide microarrays.

    Science.gov (United States)

    Turner, Helen C; Budak, Murat T; Akinci, M A Murat; Wolosin, J Mario

    2007-05-01

    To determine global mRNA expression levels in corneal and conjunctival epithelia and identify transcripts that exhibit preferential tissue expression. cDNA samples derived from human conjunctival and corneal epithelia were hybridized in three independent experiments to a commercial oligonucleotide array representing more than 22,000 transcripts. The resultant signal intensities and microarray software transcript present/absent calls were used in conjunction with the local pooled error (LPE) statistical method to identify transcripts that are preferentially or exclusively expressed in one of the two tissues at significant levels (expression >1% of the beta-actin level). EASE (Expression Analysis Systematic Explorer software) was used to identify biological systems comparatively overrepresented in either epithelium. Immuno-, and cytohistochemistry was performed to validate or expand on selected results of interest. The analysis identified 332 preferential and 93 exclusive significant corneal epithelial transcripts. The corresponding numbers of conjunctival epithelium transcripts were 592 and 211, respectively. The overrepresented biological processes in the cornea were related to cell adhesion and oxiredox equilibria and cytoprotection activities. In the conjunctiva, the biological processes that were most prominent were related to innate immunity and melanogenesis. Immunohistochemistry for antigen-presenting cells and melanocytes was consistent with these gene signatures. The transcript comparison identified a substantial number of genes that have either not been identified previously or are not known to be highly expressed in these two epithelia, including testican-1, ECM1, formin, CRTAC1, and NQO1 in the cornea and, in the conjunctiva, sPLA(2)-IIA, lipocalin 2, IGFBP3, multiple MCH class II proteins, and the Na-Pi cotransporter type IIb. Comparative gene expression profiling leads to the identification of many biological processes and previously unknown genes that

  19. Associations between stereotype awareness, childhood trauma and psychopathology: a study in people with psychosis, their siblings and controls.

    Directory of Open Access Journals (Sweden)

    Catherine van Zelst

    Full Text Available Stereotype awareness--or an individual's perception of the degree to which negative beliefs or stereotypes are held by the public--is an important factor mediating public stigma, self-stigma and their negative consequences. Research is required to assess how individuals become more sensitive to perceive stereotypes, pointing the way to therapeutic options to reduce its negative effects and increase stigma resilience. Because perception and interpretation can be guided by belief systems, and childhood trauma (CT is reported to impact such beliefs, CT is explored in relation to stereotype awareness (SA in persons with psychosis, their siblings and controls.Data from the GROUP project (Genetic Risk and Outcome of Psychosis were analyzed. SA was measured by devaluation scales which assess a respondent's perception of the degree to which stereotypes about people with mental illness and about their families are held by the public. CT was measured using the Childhood Trauma Questionnaire (short form.In patients, symptoms of disorganization and emotional distress were associated with SA about people with mental illness. In siblings, schizotypal features were associated with both types of SA (more schizotypy = more SA. In both patients and siblings, CT was associated with both types of SA (more CT = more SA, independent of symptoms (patients or schizotypy (siblings.CT in people with psychosis and their siblings may sensitize to SA. Thus, CT may not only impact on risk for illness onset, it may also increase SA associated with mental illness, potentially interfering with the recovery process. CT-induced SA may indicate a heightened sensitivity to threat, which may also impact psychopathology.

  20. Carbonated water (CW) process waste reuse for ammonium-uranyl-carbonate (AUC) production and its gains on the environmental, economic and social aspects

    Energy Technology Data Exchange (ETDEWEB)

    Carnaval, Joao Paulo R.; Santos, Rafael D. dos; Barbosa, Rodrigo A.; Lauer, Sergio, E-mail: joaocarnaval@inb.gov.br, E-mail: rafaelsantos@inb.gov.br, E-mail: rodrigobarbosa@inb.gov.br, E-mail: lauer@inb.gov.br [Industias Nucleares do Brasil S.A. (INB), Resende, RJ (Brazil)

    2013-07-01

    In the INB nuclear fuel cycle, the pellets production is based on UO{sub 2} powder made by AUC (Ammonium-Uranyl-Carbonate) route. AUC formation occurs by fluidising of UF{sub 6}, NH{sub 3} and CO{sub 2} in a vase containing usually pure water, and this exothermal reaction has AUC as direct product. The mass formed is filtered, washed with CW, washed again with methano solution, dried with air and conducted to the fluidized bed furnace, to be converted to UO{sub 2} powder. At this point, the dried AUC decompounds to UO{sub 3}, NH{sub 3} and C0{sub 2}, these 2 gases are absorbed at the gases washer, formin go the carbonated water (CW), whit is basically a (NH{sub 4}){sub 2}CO{sub 3} solution. The UO{sub 2+x} is reduced and stabilized to UO{sub 2} powder, which is conducted to pellets production. During the process, a considerable amount of this aqueous waste is generated and goes for effluent treatment. After that, the solution is sent for spray-dryer for power formation, and stock. This treatment demands equipment, energy and time, representing considerable costs of the company beyond the human risks involved on the drying step. The purpose of this work is to present a study of the carbonated water use as substitute of pure water in the AUC formation step. At this point, tests were made varying the CW loads for the AUC precipitation, and the control was made by the UO{sub 2} powder properties. The carbonated water used for AUC precipitation has been tested at several levels and the results has demonstrated full viability to become a definitive process step (INB, Resende site). It has been demonstrated the great resources economy caused by the waste reuse and the guarantee product quality. This represents such an environmental gain and also economic and social aspects got improved. (author)

  1. Peptide–polymer ligands for a tandem WW-domain, an adaptive multivalent protein–protein interaction: lessons on the thermodynamic fitness of flexible ligands

    Directory of Open Access Journals (Sweden)

    Katharina Koschek

    2015-05-01

    Full Text Available Three polymers, poly(N-(2-hydroxypropylmethacrylamide (pHPMA, hyperbranched polyglycerol (hPG, and dextran were investigated as carriers for multivalent ligands targeting the adaptive tandem WW-domain of formin-binding protein (FBP21. Polymer carriers were conjugated with 3–9 copies of the proline-rich decapeptide GPPPRGPPPR-NH2 (P1. Binding of the obtained peptide–polymer conjugates to the tandem WW-domain was investigated employing isothermal titration calorimetry (ITC to determine the binding affinity, the enthalpic and entropic contributions to free binding energy, and the stoichiometry of binding for all peptide–polymer conjugates. Binding affinities of all multivalent ligands were in the µM range, strongly amplified compared to the monovalent ligand P1 with a KD > 1 mM. In addition, concise differences were observed, pHPMA and hPG carriers showed moderate affinity and bound 2.3–2.8 peptides per protein binding site resulting in the formation of aggregates. Dextran-based conjugates displayed affinities down to 1.2 µM, forming complexes with low stoichiometry, and no precipitation. Experimental results were compared with parameters obtained from molecular dynamics simulations in order to understand the observed differences between the three carrier materials. In summary, the more rigid and condensed peptide–polymer conjugates based on the dextran scaffold seem to be superior to induce multivalent binding and to increase affinity, while the more flexible and dendritic polymers, pHPMA and hPG are suitable to induce crosslinking upon binding.

  2. Folding kinetics of WW domains with the united residue force field for bridging microscopic motions and experimental measurements.

    Science.gov (United States)

    Zhou, Rui; Maisuradze, Gia G; Suñol, David; Todorovski, Toni; Macias, Maria J; Xiao, Yi; Scheraga, Harold A; Czaplewski, Cezary; Liwo, Adam

    2014-12-23

    To demonstrate the utility of the coarse-grained united-residue (UNRES) force field to compare experimental and computed kinetic data for folding proteins, we have performed long-time millisecond-timescale canonical Langevin molecular dynamics simulations of the triple β-strand from the Formin binding protein 28 WW domain and six nonnatural variants, using UNRES. The results have been compared with available experimental data in both a qualitative and a quantitative manner. Complexities of the folding pathways, which cannot be determined experimentally, were revealed. The folding mechanisms obtained from the simulated folding kinetics are in agreement with experimental results, with a few discrepancies for which we have accounted. The origins of single- and double-exponential kinetics and their correlations with two- and three-state folding scenarios are shown to be related to the relative barrier heights between the various states. The rate constants obtained from time profiles of the fractions of the native, intermediate, and unfolded structures, and the kinetic equations fitted to them, correlate with the experimental values; however, they are about three orders of magnitude larger than the experimental ones for most of the systems. These differences are in agreement with the timescale extension derived by scaling down the friction of water and averaging out the fast degrees of freedom when passing from all-atom to a coarse-grained representation. Our results indicate that the UNRES force field can provide accurate predictions of folding kinetics of these WW domains, often used as models for the study of the mechanisms of proein folding.

  3. Molecular and Cellular mechanisms of Shigella flexneri dissemination

    Directory of Open Access Journals (Sweden)

    Herve eAgaisse

    2016-03-01

    Full Text Available The intracellular pathogen Shigella flexneri is the causative agent of bacillary dysentery in humans. The disease is characterized by bacterial invasion of intestinal cells, dissemination within the colonic epithelium through direct spread from cell to cell, and massive inflammation of the intestinal mucosa. Here, we review the mechanisms supporting S. flexneri dissemination. The dissemination process primarily relies on actin assembly at the bacterial pole, which propels the pathogen throughout the cytosol of primary infected cells. Polar actin assembly is supported by polar expression of the bacterial autotransporter family member IcsA, which recruits the N-WASP/ARP2/3 actin assembly machinery. As motile bacteria encounter cell-cell contacts, they form plasma membrane protrusions that project into adjacent cells. In addition to the ARP2/3-dependent actin assembly machinery, protrusion formation relies on formins and myosins. The resolution of protrusions into vacuoles occurs through the collapse of the protrusion neck, leading to the formation of an intermediate membrane-bound compartment termed vacuole-like protrusions (VLPs. VLP formation requires tyrosine kinase and phosphoinositide signaling in protrusions, which relies on the integrity of the bacterial type 3 secretion system (T3SS. The T3SS is also required for escaping double membrane vacuoles through the activity of the T3SS translocases IpaB and IpaC, and the effector proteins VirA and IcsB. Numerous factors supporting envelope biogenesis contribute to IcsA exposure and maintenance at the bacterial pole, including LPS synthesis, membrane proteases, and periplasmic chaperones. Although less characterized, the assembly and function of the T3SS in the context of bacterial dissemination also relies on factors supporting envelope biogenesis. Finally, the dissemination process requires the adaptation of the pathogen to various cellular compartments through transcriptional and post

  4. Mobilization of HIV spread by diaphanous 2 dependent filopodia in infected dendritic cells.

    Directory of Open Access Journals (Sweden)

    Anupriya Aggarwal

    Full Text Available Paramount to the success of persistent viral infection is the ability of viruses to navigate hostile environments en route to future targets. In response to such obstacles, many viruses have developed the ability of establishing actin rich-membrane bridges to aid in future infections. Herein through dynamic imaging of HIV infected dendritic cells, we have observed how viral high-jacking of the actin/membrane network facilitates one of the most efficient forms of HIV spread. Within infected DC, viral egress is coupled to viral filopodia formation, with more than 90% of filopodia bearing immature HIV on their tips at extensions of 10 to 20 µm. Live imaging showed HIV filopodia routinely pivoting at their base, and projecting HIV virions at µm.sec⁻¹ along repetitive arc trajectories. HIV filopodial dynamics lead to up to 800 DC to CD4 T cell contacts per hour, with selection of T cells culminating in multiple filopodia tethering and converging to envelope the CD4 T-cell membrane with budding HIV particles. Long viral filopodial formation was dependent on the formin diaphanous 2 (Diaph2, and not a dominant Arp2/3 filopodial pathway often associated with pathogenic actin polymerization. Manipulation of HIV Nef reduced HIV transfer 25-fold by reducing viral filopodia frequency, supporting the potency of DC HIV transfer was dependent on viral filopodia abundance. Thus our observations show HIV corrupts DC to CD4 T cell interactions by physically embedding at the leading edge contacts of long DC filopodial networks.

  5. Conserved cis-regulatory regions in a large genomic landscape control SHH and BMP-regulated Gremlin1 expression in mouse limb buds

    Directory of Open Access Journals (Sweden)

    Zuniga Aimée

    2012-08-01

    Full Text Available Abstract Background Mouse limb bud is a prime model to study the regulatory interactions that control vertebrate organogenesis. Major aspects of limb bud development are controlled by feedback loops that define a self-regulatory signalling system. The SHH/GREM1/AER-FGF feedback loop forms the core of this signalling system that operates between the posterior mesenchymal organiser and the ectodermal signalling centre. The BMP antagonist Gremlin1 (GREM1 is a critical node in this system, whose dynamic expression is controlled by BMP, SHH, and FGF signalling and key to normal progression of limb bud development. Previous analysis identified a distant cis-regulatory landscape within the neighbouring Formin1 (Fmn1 locus that is required for Grem1 expression, reminiscent of the genomic landscapes controlling HoxD and Shh expression in limb buds. Results Three highly conserved regions (HMCO1-3 were identified within the previously defined critical genomic region and tested for their ability to regulate Grem1 expression in mouse limb buds. Using a combination of BAC and conventional transgenic approaches, a 9 kb region located ~70 kb downstream of the Grem1 transcription unit was identified. This region, termed Grem1 Regulatory Sequence 1 (GRS1, is able to recapitulate major aspects of Grem1 expression, as it drives expression of a LacZ reporter into the posterior and, to a lesser extent, in the distal-anterior mesenchyme. Crossing the GRS1 transgene into embryos with alterations in the SHH and BMP pathways established that GRS1 depends on SHH and is modulated by BMP signalling, i.e. integrates inputs from these pathways. Chromatin immunoprecipitation revealed interaction of endogenous GLI3 proteins with the core cis-regulatory elements in the GRS1 region. As GLI3 is a mediator of SHH signal transduction, these results indicated that SHH directly controls Grem1 expression through the GRS1 region. Finally, all cis-regulatory regions within the Grem1

  6. The MARVEL domain protein Nce102 regulates actin organization and invasive growth of Candida albicans.

    Science.gov (United States)

    Douglas, Lois M; Wang, Hong X; Konopka, James B

    2013-11-26

    Invasive growth of the fungal pathogen Candida albicans into tissues promotes disseminated infections in humans. The plasma membrane is essential for pathogenesis because this important barrier mediates morphogenesis and invasive growth, as well as secretion of virulence factors, cell wall synthesis, nutrient import, and other processes. Previous studies showed that the Sur7 tetraspan protein that localizes to MCC (membrane compartment occupied by Can1)/eisosome subdomains of the plasma membrane regulates a broad range of key functions, including cell wall synthesis, morphogenesis, and resistance to copper. Therefore, a distinct tetraspan protein found in MCC/eisosomes, Nce102, was investigated. Nce102 belongs to the MARVEL domain protein family, which is implicated in regulating membrane structure and function. Deletion of NCE102 did not cause the broad defects seen in sur7Δ cells. Instead, the nce102Δ mutant displayed a unique phenotype in that it was defective in forming hyphae and invading low concentrations of agar but could invade well in higher agar concentrations. This phenotype was likely due to a defect in actin organization that was observed by phalloidin staining. In support of this, the invasive growth defect of a bni1Δ mutant that mislocalizes actin due to lack of the Bni1 formin was also reversed at high agar concentrations. This suggests that a denser matrix provides a signal that compensates for the actin defects. The nce102Δ mutant displayed decreased virulence and formed abnormal hyphae in mice. These studies identify novel ways that Nce102 and the physical environment surrounding C. albicans regulate morphogenesis and pathogenesis. The plasma membrane promotes virulence of the human fungal pathogen Candida albicans by acting as a protective barrier around the cell and mediating dynamic activities, such as morphogenesis, cell wall synthesis, secretion of virulence factors, and nutrient uptake. To better understand how the plasma membrane

  7. Bovine Vaccinia in dairy cattle and suspicion of vesicular disease on milkers in Brazil

    Directory of Open Access Journals (Sweden)

    Thaís Garcia da Silva

    2018-05-01

    Full Text Available ABSTRACT: Bovine vaccinia (BV is a vesicular disease induced by the Vaccinia virus (VACV that affects milk production and is an occupational zoonosis. This research had the following objectives: (i detection of VACV by qPCR in cattle with clinical suspicion of vesicular disease; (ii symptoms characterization in animals and milkers with clinical suspicion of the disease and virus detection in humans; and (iii identification of risk factors for infections of VACV in herds from several Brazilian states. A total of 471 bovine epithelial samples from dairy farms, in 15 Brazilian states, were evaluated between 2007 and 2012. The samples were tested by quantitative PCR (qPCR using SYBR Green® reagents, validated with a lower limit of detection of 100 TCID50/50µL (1.7x100 viral particles, and 45.1% of VACV positive samples were detected. Using official forms for epidemiological investigation (FORM-IN, the risk factors for VACV infections in cattle were determined to be farms with a lack of technological facilities (P=0.029 and the presence of rodents (P=0.001. There was an effect of seasonality in cattle with a higher occurrence of BV during the dry season. A total of 420 epidemiological questionnaires were applied at public health care centers, where 100% of the milkers had vesicular lesions on their hands (98.1% and on their arms (6.9%. The most frequent clinical symptoms in humans were: local swelling (74.2%, headache (20.7%, fever (10.4% and inguinal lymphadenopathy (74.2%. Only 19.98% of milkers aged between 39 and 58 years were seroreactive to VACV and were immunized with the human anti-smallpox vaccine. There was an increase in the frequency of BV in older individuals due to their natural decrease in specific immunity. It has been shown that the implementation of zootechnical management techniques and health planning are important for the prevention of BV in animals and humans.

  8. Permanent-magnet energy spectrometer for electron beams from radiotherapy accelerators

    Energy Technology Data Exchange (ETDEWEB)

    McLaughlin, David J.; Shikhaliev, Polad M.; Matthews, Kenneth L. [Department of Physics and Astronomy, Louisiana State University, 202 Nicholson Hall, Baton Rouge, Louisiana 70803-4001 (United States); Hogstrom, Kenneth R., E-mail: hogstrom@lsu.edu; Carver, Robert L.; Gibbons, John P. [Mary Bird Perkins Cancer Center, 4950 Essen Lane, Baton Rouge, Louisiana 70809-3482 and Department of Physics and Astronomy, Louisiana State University, 202 Nicholson Hall, Baton Rouge, Louisiana 70803-4001 (United States); Clarke, Taylor; Henderson, Alexander; Liang, Edison P. [Physics and Astronomy Department, Rice University, 6100 Main MS-61, Houston, Texas 77005-1827 (United States)

    2015-09-15

    Purpose: The purpose of this work was to adapt a lightweight, permanent magnet electron energy spectrometer for the measurement of energy spectra of therapeutic electron beams. Methods: An irradiation geometry and measurement technique were developed for an approximately 0.54-T, permanent dipole magnet spectrometer to produce suitable latent images on computed radiography (CR) phosphor strips. Dual-pinhole electron collimators created a 0.318-cm diameter, approximately parallel beam incident on the spectrometer and an appropriate dose rate at the image plane (CR strip location). X-ray background in the latent image, reduced by a 7.62-cm thick lead block between the pinhole collimators, was removed using a fitting technique. Theoretical energy-dependent detector response functions (DRFs) were used in an iterative technique to transform CR strip net mean dose profiles into energy spectra on central axis at the entrance to the spectrometer. These spectra were transformed to spectra at 95-cm source to collimator distance (SCD) by correcting for the energy dependence of electron scatter. The spectrometer was calibrated by comparing peak mean positions in the net mean dose profiles, initially to peak mean energies determined from the practical range of central-axis percent depth-dose (%DD) curves, and then to peak mean energies that accounted for how the collimation modified the energy spectra (recalibration). The utility of the spectrometer was demonstrated by measuring the energy spectra for the seven electron beams (7–20 MeV) of an Elekta Infinity radiotherapy accelerator. Results: Plots of DRF illustrated their dependence on energy and position in the imaging plane. Approximately 15 iterations solved for the energy spectra at the spectrometer entrance from the measured net mean dose profiles. Transforming those spectra into ones at 95-cm SCD increased the low energy tail of the spectra, while correspondingly decreasing the peaks and shifting them to slightly lower

  9. Accurate Holdup Calculations with Predictive Modeling & Data Integration

    Energy Technology Data Exchange (ETDEWEB)

    Azmy, Yousry [North Carolina State Univ., Raleigh, NC (United States). Dept. of Nuclear Engineering; Cacuci, Dan [Univ. of South Carolina, Columbia, SC (United States). Dept. of Mechanical Engineering

    2017-04-03

    Bayes’ Theorem, one must have a model y(x) that maps the state variables x (the solution in this case) to the measurements y. In this case, the unknown state variables are the configuration and composition of the heldup SNM. The measurements are the detector readings. Thus, the natural model is neutral-particle radiation transport where a wealth of computational tools exists for performing these simulations accurately and efficiently. The combination of predictive model and Bayesian inference forms the Data Integration with Modeled Predictions (DIMP) method that serves as foundation for this project. The cost functional describing the model-to-data misfit is computed via a norm created by the inverse of the covariance matrix of the model parameters and responses. Since the model y(x) for the holdup problem is nonlinear, a nonlinear optimization on Q is conducted via Newton-type iterative methods to find the optimal values of the model parameters x. This project comprised a collaboration between NC State University (NCSU), the University of South Carolina (USC), and Oak Ridge National Laboratory (ORNL). The project was originally proposed in seven main tasks with an eighth contingency task to be performed if time and funding permitted; in fact time did not permit commencement of the contingency task and it was not performed. The remaining tasks involved holdup analysis with gamma detection strategies and separately with neutrons based on coincidence counting. Early in the project, and upon consultation with experts in coincidence counting it became evident that this approach is not viable for holdup applications and this task was replaced with an alternative, but valuable investigation that was carried out by the USC partner. Nevertheless, the experimental 4 measurements at ORNL of both gamma and neutron sources for the purpose of constructing Detector Response Functions (DRFs) with the associated uncertainties were indeed completed.

  10. Permanent-magnet energy spectrometer for electron beams from radiotherapy accelerators.

    Science.gov (United States)

    McLaughlin, David J; Hogstrom, Kenneth R; Carver, Robert L; Gibbons, John P; Shikhaliev, Polad M; Matthews, Kenneth L; Clarke, Taylor; Henderson, Alexander; Liang, Edison P

    2015-09-01

    The purpose of this work was to adapt a lightweight, permanent magnet electron energy spectrometer for the measurement of energy spectra of therapeutic electron beams. An irradiation geometry and measurement technique were developed for an approximately 0.54-T, permanent dipole magnet spectrometer to produce suitable latent images on computed radiography (CR) phosphor strips. Dual-pinhole electron collimators created a 0.318-cm diameter, approximately parallel beam incident on the spectrometer and an appropriate dose rate at the image plane (CR strip location). X-ray background in the latent image, reduced by a 7.62-cm thick lead block between the pinhole collimators, was removed using a fitting technique. Theoretical energy-dependent detector response functions (DRFs) were used in an iterative technique to transform CR strip net mean dose profiles into energy spectra on central axis at the entrance to the spectrometer. These spectra were transformed to spectra at 95-cm source to collimator distance (SCD) by correcting for the energy dependence of electron scatter. The spectrometer was calibrated by comparing peak mean positions in the net mean dose profiles, initially to peak mean energies determined from the practical range of central-axis percent depth-dose (%DD) curves, and then to peak mean energies that accounted for how the collimation modified the energy spectra (recalibration). The utility of the spectrometer was demonstrated by measuring the energy spectra for the seven electron beams (7-20 MeV) of an Elekta Infinity radiotherapy accelerator. Plots of DRF illustrated their dependence on energy and position in the imaging plane. Approximately 15 iterations solved for the energy spectra at the spectrometer entrance from the measured net mean dose profiles. Transforming those spectra into ones at 95-cm SCD increased the low energy tail of the spectra, while correspondingly decreasing the peaks and shifting them to slightly lower energies. Energy calibration

  11. A systematic review of structured versus non-structured breastfeeding programmes to support the initiation and duration of exclusive breastfeeding in acute and primary healthcare settings.

    Science.gov (United States)

    Beake, Sarah; Pellowe, Carol; Dykes, Fiona; Schmied, Virginia; Bick, Debra

    2011-01-01

    Joanna Briggs Institute. Two independent reviewers conducted critical appraisal and data extraction. Twenty-six articles were included; one randomised controlled trial, two non randomised trials, one cross-sectional study, five systematic reviews, 15 cohort studies and two descriptive studies. Due to the poor quality of evidence presented and clinical and methodological heterogeneity of study designs, including definitions of breastfeeding and duration of follow-up, it was not possible to combine studies or individual outcomes in meta-analyses, therefore findings are presented in a narrative form.In most studies the structured programme of interest reflected some or all of the Baby Friendly Hospital Initiative 'Ten Steps'. Most studies found a statistically significant improvement in initiation of breastfeeding following introduction of a structured breastfeeding programme, although effect sizes varied widely.The impact of introducing a structured programme on the duration of exclusive breastfeeding and duration of any breastfeeding was also evident, although not all studies found statistically significant differences. At hospital discharge or within the first week post-birth, implementation of a structured programme appeared to increase duration of exclusive breastfeeding and the duration of any breastfeeding compared with usual care. After hospital discharge and up to six months post-birth, use of structured programmes also appeared to support continued duration of exclusive and any breastfeeding although differences in outcomes were not reported across all included studies. At six months, three of five studies which included data on longer-term outcomes showed women were statistically significantly more likely to be exclusively breastfeeding. Only one of these studies compared outcomes following implementation of BFHI. Despite the poor overall quality of studies, structured programmes, regardless of content, compared with standard care appear to influence the uptake

  12. FOREWORD Nanomaterials science Nanomaterials science

    Science.gov (United States)

    Rohrer, Heinrich

    2010-10-01

    nanotubes of various lengths in complex micro- and nano-electronic circuits, however, they have to be grown at given positions, which is still problematic. Another example concerns the assembly scenario for electronics, components like sensors, actuators, and nano-systems. Macromolecular chemistry is producing highly functional macromolecules, but, eventually, they have to be produced and assembled at given positions. Shaping materials to a given nanosize and formIn the field of micro- and nanoelectronics, shaping semiconductors, many oxides, and selected metals down to 20-100 nm dimensions is standard. In nanomechanics, however, other materials might be more appropriate and better suited for a given task. In other cases, finishing procedures might be impossible or too time-consuming for large numbers of them. Components for counting electrons—more elegant and smaller than today's single-electron transistors—or adjustable holes for counting atoms and molecules will eventually be badly needed because of the 1/√N fluctuations in the properties and measurements at a small N, for example, N dopants in nanosize transistors or N electrons in very short current pulses. Bistable componentsBistable components, which do not require electrical currents, are aimed at reducing local energy dissipation and faster startup of personal computers. Magnetoresistive and ferroelectric random access memory (MRAMS and FRAMS) devices are the first attempts to use them in circuits. I am not aware of reported switching times that are considerably faster than a few nanoseconds, as required in today's storage. This is too slow for memory and much too slow for possible logic devices based on two-terminal bistable components. Bistable molecules, a mechanical switch, might be a valid and sufficiently fast alternative, certainly with all the challenges mentioned above. I have mentioned just a few obvious examples of the involvement of materials science in the new world of nanodimensions. However

  13. Nanomaterials science

    Directory of Open Access Journals (Sweden)

    Heinrich Rohrer

    2010-01-01

    carbon nanotubes of various lengths in complex micro- and nano-electronic circuits, however, they have to be grown at given positions, which is still problematic. Another example concerns the assembly scenario for electronics, components like sensors, actuators, and nano-systems. Macromolecular chemistry is producing highly functional macromolecules, but, eventually, they have to be produced and assembled at given positions.Shaping materials to a given nanosize and formIn the field of micro- and nanoelectronics, shaping semiconductors, many oxides, and selected metals down to 20–100 nm dimensions is standard. In nanomechanics, however, other materials might be more appropriate and better suited for a given task. In other cases, finishing procedures might be impossible or too time-consuming for large numbers of them. Components for counting electrons—more elegant and smaller than today's single-electron transistors—or adjustable holes for counting atoms and molecules will eventually be badly needed because of the 1/√N fluctuations in the properties and measurements at a small N, for example, N dopants in nanosize transistors or N electrons in very short current pulses.Bistable componentsBistable components, which do not require electrical currents, are aimed at reducing local energy dissipation and faster startup of personal computers. Magnetoresistive and ferroelectric random access memory (MRAMS and FRAMS devices are the first attempts to use them in circuits. I am not aware of reported switching times that are considerably faster than a few nanoseconds, as required in today's storage. This is too slow for memory and much too slow for possible logic devices based on two-terminal bistable components. Bistable molecules, a mechanical switch, might be a valid and sufficiently fast alternative, certainly with all the challenges mentioned above.I have mentioned just a few obvious examples of the involvement of materials science in the new world of nanodimensions