Simian Immunodeficiency Virus and Human Immunodeficiency Virus Type 1 Nef Proteins Show Distinct Patterns and Mechanisms of Src Kinase Activation

Science.gov (United States)

Greenway, Alison L.; Dutartre, Hélène; Allen, Kelly; McPhee, Dale A.; Olive, Daniel; Collette, Yves

1999-01-01

The nef gene from human and simian immunodeficiency viruses (HIV and SIV) regulates cell function and viral replication, possibly through binding of the nef product to cellular proteins, including Src family tyrosine kinases. We show here that the Nef protein encoded by SIVmac239 interacts with and also activates the human Src kinases Lck and Hck. This is in direct contrast to the inhibitory effect of HIV type 1 (HIV-1) Nef on Lck catalytic activity. Unexpectedly, however, the interaction of SIV Nef with human Lck or Hck is not mediated via its consensus proline motif, which is known to mediate HIV-1 Nef binding to Src homology 3 (SH3) domains, and various experimental analyses failed to show significant interaction of SIV Nef with the SH3 domain of either kinase. Instead, SIV Nef can bind Lck and Hck SH2 domains, and its N-terminal 50 amino acid residues are sufficient for Src kinase binding and activation. Our results provide evidence for multiple mechanisms by which Nef binds to and regulates Src kinases. PMID:10364375

Aging of SRC liquids

Science.gov (United States)

Hara, T.; Jones, L.; Tewari, K. C.; Li, N. C.

1981-02-01

The viscosity of SRC-LL liquid increases when subjected to accelerated aging by bubbling oxygen in the presence of copper strip at 62°C. Precipitates are formed and can be separated from the aged liquid by Soxhlet extraction with pentane. A 30-70 blend of SRC-I with SRC-LL was subjected to oxygen aging in the absence of copper, and the viscosity increased dramatically after 6 days at 62°. The content of preasphaltene and its molecular size increase with time of aging, accompanied by decrease of asphaltene and pentane-soluble contents. For the preasphaltene fraction on aging, gel permeation chromatography shows formation of larger particles. ESR experiments show that with oxygen aging, spin concentration in the preasphaltene fraction decreases. Perhaps some semiquinone, together with di- and tri-substituted phenoxy radicals, generated by oxygen aging of the coal liquid, interact with the free radicals already present in coal to yield larger particles and reduce free radical concentration. We are currently using the very high-field (600-MHz) NMR spectrometer at Mellon Institute to determine changes in structural parameters before and after aging of SRC-II and its chromatographically separated fractions.

Report on evaluation for SRC-2 coal liquefaction project; SRC-II sekitan ekika project hyoka sagyo hokokusho

Energy Technology Data Exchange (ETDEWEB)

NONE

1979-10-01

Among the EDS, H-Coal and SRC-2 lined up in the coal liquefaction project of the U.S., the SRC is aimed at producing 6,000 t/day as a module for a 30,000 t/day commercial plant. They expect Ash contents (iron, sulfur, etc.) in coal without basically using catalysts. The products are applicable to fuels for electricity and gas for the moment. In the element technology, there are some problems in manufacturing hydrogen by gasification of residuals as well as in slurry systems, reaction towers, etc.. In the 30,000 ton commercial plant, the coal-liquefied oil costs $19.89/bbl (price as of 1978) assuming coal costs $29.47; therefore, the feasibility is strong as a substitute for petroleum. Japan's share for the required funds will be 86.8 billion yen (if 250 yen per dollar). Since the kinds of coal are conceivably increased in number through the improvement of the process, the Pacific rim countries and these which lie on the Indian Ocean are assumed to be the major coal producing countries for Japan. The stability in storage of coal-liquefied oil is experimentally excellent, as is the compatibility with petroleum products for example. Great results can be expected in the technical know-how and the spread of element technology for Japan. (NEDO)

Report on evaluation for SRC-2 coal liquefaction project; SRC-II sekitan ekika project hyoka sagyo hokokusho

Energy Technology Data Exchange (ETDEWEB)

NONE

1979-10-01

Among the EDS, H-Coal and SRC-2 lined up in the coal liquefaction project of the U.S., the SRC is aimed at producing 6,000 t/day as a module for a 30,000 t/day commercial plant. They expect Ash contents (iron, sulfur, etc.) in coal without basically using catalysts. The products are applicable to fuels for electricity and gas for the moment. In the element technology, there are some problems in manufacturing hydrogen by gasification of residuals as well as in slurry systems, reaction towers, etc.. In the 30,000 ton commercial plant, the coal-liquefied oil costs $19.89/bbl (price as of 1978) assuming coal costs $29.47; therefore, the feasibility is strong as a substitute for petroleum. Japan's share for the required funds will be 86.8 billion yen (if 250 yen per dollar). Since the kinds of coal are conceivably increased in number through the improvement of the process, the Pacific rim countries and these which lie on the Indian Ocean are assumed to be the major coal producing countries for Japan. The stability in storage of coal-liquefied oil is experimentally excellent, as is the compatibility with petroleum products for example. Great results can be expected in the technical know-how and the spread of element technology for Japan. (NEDO)

Aladdin: Transforming science at SRC

Science.gov (United States)

Bisognano, J.; Bissen, M.; Green, M.; Jacobs, K.; Moore, C.; Olson, E.; Severson, M.; Wehlitz, R.

2011-09-01

The Synchrotron Radiation Center (SRC) is dedicated to enabling of innovative research using IR, ultraviolet, and soft X-ray synchrotron radiation. It delivers beam time with high reliability (99%) and continues to improve the Aladdin storage ring complex. A lower emittance tuning has been commissioned to support a microfocus capability. SRC successfully installed an APPLE II undulator providing elliptically polarized light with lattice compensation for flexible scanning. Installation of a new IR beamline at SRC is providing synchrotron chemical imaging with unprecedented structural and chemical information, simultaneously. In addition, SRC has established a strong education and outreach program to bring the knowledge and power of light source science to a wider national community. It is moving forward into the future by developing a new micro focus beamline producing a diffraction-limited focus of about 500 nm at 22 eV, proposing an additional diffraction-limited chemical imaging beamline, and advancing the Wisconsin Free Electron Laser (WiFEL) concept.

Aladdin: Transforming science at SRC

International Nuclear Information System (INIS)

Bisognano, J.; Bissen, M.; Green, M.; Jacobs, K.; Moore, C.; Olson, E.; Severson, M.; Wehlitz, R.

2011-01-01

The Synchrotron Radiation Center (SRC) is dedicated to enabling of innovative research using IR, ultraviolet, and soft X-ray synchrotron radiation. It delivers beam time with high reliability (99%) and continues to improve the Aladdin storage ring complex. A lower emittance tuning has been commissioned to support a microfocus capability. SRC successfully installed an APPLE II undulator providing elliptically polarized light with lattice compensation for flexible scanning. Installation of a new IR beamline at SRC is providing synchrotron chemical imaging with unprecedented structural and chemical information, simultaneously. In addition, SRC has established a strong education and outreach program to bring the knowledge and power of light source science to a wider national community. It is moving forward into the future by developing a new micro focus beamline producing a diffraction-limited focus of about 500 nm at 22 eV, proposing an additional diffraction-limited chemical imaging beamline, and advancing the Wisconsin Free Electron Laser (WiFEL) concept.

Tyrosine phosphorylation of LRP6 by Src and Fer inhibits Wnt/β-catenin signalling

Science.gov (United States)

Chen, Qing; Su, Yi; Wesslowski, Janine; Hagemann, Anja I; Ramialison, Mirana; Wittbrodt, Joachim; Scholpp, Steffen; Davidson, Gary

2014-01-01

Low-density lipoprotein receptor-related proteins 5 and 6 (LRP5/6) function as transmembrane receptors to transduce Wnt signals. A key mechanism for signalling is Wnt-induced serine/threonine phosphorylation at conserved PPPSPxS motifs in the LRP6 cytoplasmic domain, which promotes pathway activation. Conserved tyrosine residues are positioned close to all PPPSPxS motifs, which suggests they have a functional significance. Using a cell culture-based cDNA expression screen, we identified the non-receptor tyrosine kinases Src and Fer as novel LRP6 modifiers. Both Src and Fer associate with LRP6 and phosphorylate LRP6 directly. In contrast to the known PPPSPxS Ser/Thr kinases, tyrosine phosphorylation by Src and Fer negatively regulates LRP6-Wnt signalling. Epistatically, they function upstream of β-catenin to inhibit signalling and in agreement with a negative role in regulating LRP6, MEF cells lacking these kinases show enhanced Wnt signalling. Wnt3a treatment of cells enhances tyrosine phosphorylation of endogenous LRP6 and, mechanistically, Src reduces cell surface LRP6 levels and disrupts LRP6 signalosome formation. Interestingly, CK1γ inhibits Fer-induced LRP6 phosphorylation, suggesting a mechanism whereby CK1γ acts to de-represses inhibitory LRP6 tyrosine phosphorylation. We propose that LRP6 tyrosine phosphorylation by Src and Fer serves a negative regulatory function to prevent over-activation of Wnt signalling at the level of the Wnt receptor, LRP6. Subject Categories Membrane & Intracellular Transport; Post-translational Modifications, Proteolysis & Proteomics PMID:25391905

Differential sensitivity of Src-family kinases to activation by SH3 domain displacement.

Directory of Open Access Journals (Sweden)

Jamie A Moroco

Full Text Available Src-family kinases (SFKs are non-receptor protein-tyrosine kinases involved in a variety of signaling pathways in virtually every cell type. The SFKs share a common negative regulatory mechanism that involves intramolecular interactions of the SH3 domain with the PPII helix formed by the SH2-kinase linker as well as the SH2 domain with a conserved phosphotyrosine residue in the C-terminal tail. Growing evidence suggests that individual SFKs may exhibit distinct activation mechanisms dictated by the relative strengths of these intramolecular interactions. To elucidate the role of the SH3:linker interaction in the regulation of individual SFKs, we used a synthetic SH3 domain-binding peptide (VSL12 to probe the sensitivity of downregulated c-Src, Hck, Lyn and Fyn to SH3-based activation in a kinetic kinase assay. All four SFKs responded to VSL12 binding with enhanced kinase activity, demonstrating a conserved role for SH3:linker interaction in the control of catalytic function. However, the sensitivity and extent of SH3-based activation varied over a wide range. In addition, autophosphorylation of the activation loops of c-Src and Hck did not override regulatory control by SH3:linker displacement, demonstrating that these modes of activation are independent. Our results show that despite the similarity of their downregulated conformations, individual Src-family members show diverse responses to activation by domain displacement which may reflect their adaptation to specific signaling environments in vivo.

SRC-I Project Baseline. [SRC-I demonstration project near Owensboro, Kentucky

Energy Technology Data Exchange (ETDEWEB)

None

1982-03-01

The Process Design Criteria Specification forms the basis for process design for the 6000-TPSD SRC-I Demonstration Plant. It sets forth: basic engineering data, e.g., type and size of plant, feedstocks, product specifications, and atmospheric emission and waste disposal limits; utility conditions; equipment design criteria and sparing philosophy; and estimating criteria for economic considerations. Previously the formal ICRC Document No. 0001-01-002 has been submitted to DOE and revised, as necessary, to be consistent with the SRC-I Project Baseline. Revision 6, dated 19 March 1982, 51 pages, was forwarded to DOE on 19 March 1982.

Characterization of a novel weak interaction between MUC1 and Src-SH3 using nuclear magnetic resonance spectroscopy

Energy Technology Data Exchange (ETDEWEB)

Gunasekara, Nirosha [Department of Laboratory Medicine and Pathology, University of Alberta, 5B4.21 WCM Health Science Centre, 8440-112th Street, Edmonton, Alberta, Canada T6G 2R7 (Canada); Sykes, Brian, E-mail: brian.sykes@ualberta.ca [Department of Biochemistry, 4-19B Medical Sciences Bldg., University of Alberta Edmonton, Alberta, Canada T6G 2H7 (Canada); Hugh, Judith, E-mail: judithh@ualberta.ca [Department of Laboratory Medicine and Pathology, University of Alberta, 5B4.21 WCM Health Science Centre, 8440-112th Street, Edmonton, Alberta, Canada T6G 2R7 (Canada)

2012-05-18

Highlights: Black-Right-Pointing-Pointer MUC1 binds the Src-SH3 domain potentially triggering Src dependent cell migration. Black-Right-Pointing-Pointer NMR Spectroscopy was used to monitor MUC1-CD and Src SH3 domain titrations. Black-Right-Pointing-Pointer MUC1-CD peptides bind with a low affinity (K{sub d} of 2-3 mM) to a non-canonical site. Black-Right-Pointing-Pointer Weak interactions may mediate dynamic processes like migration. Black-Right-Pointing-Pointer The MUC1-CD and Src-SH3 interaction may be a prime target to inhibit cell migration. -- Abstract: Breast cancer causes death through cancer cell migration and subsequent metastasis to distant organs. In vitro, the MUC1 mucin can mediate breast cancer cell migration by binding to intercellular adhesion molecule-1 (ICAM-1). This migration is dependent on MUC1 cytoplasmic domain (MUC1-CD) activation of the non-receptor tyrosine kinase, Src, possibly through competitive displacement of an inhibitory Src intramolecular SH3 binding. Therefore, we characterized the binding site and affinity of the MUC1-CD for Src-SH3 using multidimensional nuclear magnetic resonance (NMR) spectroscopy to monitor the titration of the {sup 15}N labeled Src-SH3 domain with synthetic native and mutant peptides of MUC1-CD. The results revealed that the dissociation constant (K{sub d}) for the interaction of the native MUC1-CD peptides and Src-SH3 domain was weak with a K{sub d} of 2-3 mM. Notably, the SH3 residues most perturbed upon peptide binding were located outside the usual hydrophobic binding cleft in a previously described alternate binding site on the Src-SH3, suggesting that MUC1-CD binds to a non-canonical site. The binding characteristics outlined here suggest that the interaction between Src-SH3 and MUC1-CD represents a novel weak electrostatic interaction of the type which is increasingly recognized as important in transient and dynamic protein complexes required for cell migration and signal transduction. As such, this

Src Induces Podoplanin Expression to Promote Cell Migration*

Science.gov (United States)

Shen, Yongquan; Chen, Chen-Shan; Ichikawa, Hitoshi; Goldberg, Gary S.

2010-01-01

Nontransformed cells can force tumor cells to assume a normal morphology and phenotype by the process of contact normalization. Transformed cells must escape this process to become invasive and malignant. However, mechanisms underlying contact normalization have not been elucidated. Here, we have identified genes that are affected by contact normalization of Src-transformed cells. Tumor cells must migrate to become invasive and malignant. Src must phosphorylate the adaptor protein Cas (Crk-associated substrate) to promote tumor cell motility. We report here that Src utilizes Cas to induce podoplanin (Pdpn) expression to promote tumor cell migration. Pdpn is a membrane-bound extracellular glycoprotein that associates with endogenous ligands to promote tumor cell migration leading to cancer invasion and metastasis. In fact, Pdpn expression accounted for a major part of the increased migration seen in Src-transformed cells. Moreover, nontransformed cells suppressed Pdpn expression in adjacent Src-transformed cells. Of >39,000 genes, Pdpn was one of only 23 genes found to be induced by transforming Src activity and suppressed by contact normalization of Src-transformed cells. In addition, we found 16 genes suppressed by Src and induced by contact normalization. These genes encode growth factor receptors, adaptor proteins, and products that have not yet been annotated and may play important roles in tumor cell growth and migration. PMID:20123990

Lack of Csk-mediated negative regulation in a unicellular SRC kinase.

Science.gov (United States)

Schultheiss, Kira P; Suga, Hiroshi; Ruiz-Trillo, Iñaki; Miller, W Todd

2012-10-16

Phosphotyrosine-based signaling plays a vital role in cellular communication in multicellular organisms. Unexpectedly, unicellular choanoflagellates (the closest phylogenetic group to metazoans) possess numbers of tyrosine kinases that are comparable to those in complex metazoans. Here, we have characterized tyrosine kinases from the filasterean Capsaspora owczarzaki, a unicellular protist representing the sister group to choanoflagellates and metazoans. Two Src-like tyrosine kinases have been identified in C. owczarzaki (CoSrc1 and CoSrc2), both of which have the arrangement of SH3, SH2, and catalytic domains seen in mammalian Src kinases. In Capsaspora cells, CoSrc1 and CoSrc2 localize to punctate structures in filopodia that may represent primordial focal adhesions. We have cloned, expressed, and purified both enzymes. CoSrc1 and CoSrc2 are active tyrosine kinases. Mammalian Src kinases are normally regulated in a reciprocal fashion by autophosphorylation in the activation loop (which increases activity) and by Csk-mediated phosphorylation of the C-terminal tail (which inhibits activity). Similar to mammalian Src kinases, the enzymatic activities of CoSrc1 and CoSrc2 are increased by autophosphorylation in the activation loop. We have identified a Csk-like kinase (CoCsk) in the genome of C. owczarzaki. We cloned, expressed, and purified CoCsk and found that it has no measurable tyrosine kinase activity. Furthermore, CoCsk does not phosphorylate or regulate CoSrc1 or CoSrc2 in cells or in vitro, and CoSrc1 and CoSrc2 are active in Capsaspora cell lysates. Thus, the function of Csk as a negative regulator of Src family kinases appears to have arisen with the emergence of metazoans.

Tyrosine Phosphorylation of the Lyn Src Homology 2 (SH2) Domain Modulates Its Binding Affinity and Specificity*

Science.gov (United States)

Jin, Lily L.; Wybenga-Groot, Leanne E.; Tong, Jiefei; Taylor, Paul; Minden, Mark D.; Trudel, Suzanne; McGlade, C. Jane; Moran, Michael F.

2015-01-01

Src homology 2 (SH2) domains are modular protein structures that bind phosphotyrosine (pY)-containing polypeptides and regulate cellular functions through protein-protein interactions. Proteomics analysis showed that the SH2 domains of Src family kinases are themselves tyrosine phosphorylated in blood system cancers, including acute myeloid leukemia, chronic lymphocytic leukemia, and multiple myeloma. Using the Src family kinase Lyn SH2 domain as a model, we found that phosphorylation at the conserved SH2 domain residue Y194 impacts the affinity and specificity of SH2 domain binding to pY-containing peptides and proteins. Analysis of the Lyn SH2 domain crystal structure supports a model wherein phosphorylation of Y194 on the EF loop modulates the binding pocket that engages amino acid side chains at the pY+2/+3 position. These data indicate another level of regulation wherein SH2-mediated protein-protein interactions are modulated by SH2 kinases and phosphatases. PMID:25587033

HSP27 phosphorylation modulates TRAIL-induced activation of Src-Akt/ERK signaling through interaction with β-arrestin2.

Science.gov (United States)

Qi, Shimei; Xin, Yinqiang; Qi, Zhilin; Xu, Yimiao; Diao, Ying; Lan, Lei; Luo, Lan; Yin, Zhimin

2014-03-01

Heat shock protein 27 (HSP27) regulates critical cellular functions such as development, differentiation, cell growth and apoptosis. A variety of stimuli induce the phosphorylation of HSP27, which affects its cellular functions. However, most previous studies focused on the role of HSP27 protein itself in apoptosis, the particular role of its phosphorylation state in signaling transduction remains largely unclear. In the present study, we reported that HSP27 phosphorylation modulated TRAIL-triggered pro-survival signaling transduction. In HeLa cells, suppression of HSP27 phosphorylation by specific inhibitor KRIBB3 or MAPKAPK2 (MK2) knockdown and by overexpression of non-phosphorylatable HSP27(3A) mutant demonstrated that hindered HSP27 phosphorylation enhanced the TRAIL-induced apoptosis. In addition, reduced HSP27 phosphorylation by KRIBB3 treatment or MK2 knockdown attenuated the TRAIL-induced activation of Akt and ERK survival signaling through suppressing the phosphorylation of Src. By overexpression of HSP27(15A) or HSP27(78/82A) phosphorylation mutant, we further showed that phosphorylation of HSP27 at serine 78/82 residues was essential to TRAIL-triggered Src-Akt/ERK signaling transduction. Co-immunoprecipitation and confocal microscopy showed that HSP27 interacted with Src and scaffolding protein β-arrestin2 in response of TRAIL stimulation and suppression of HSP27 phosphorylation apparently disrupted the TRAIL-induced interaction of HSP27 and Src or interaction of HSP27 and β-arrestin2. We further demonstrated that β-arrestin2 mediated HSP27 action on TRAIL-induced Src activation, which was achieved by recruiting signaling complex of HSP27/β-arrestin2/Src in response to TRAIL. Taken together, our study revealed that HSP27 phosphorylation modulates TRAIL-triggered activation of Src-Akt/ERK pro-survival signaling via interacting with β-arrestin2 in HeLa cells. Copyright © 2013 Elsevier Inc. All rights reserved.

The Src SH2 domain interacts dynamically with the focal adhesion kinase binding site as demonstrated by paramagnetic NMR spectroscopy.

Science.gov (United States)

Lindfors, Hanna E; Drijfhout, Jan Wouter; Ubbink, Marcellus

2012-06-01

The interaction between the tyrosine kinases Src and focal adhesion kinase (FAK) is a key step in signaling processes from focal adhesions. The phosphorylated tyrosine residue 397 in FAK is able to bind the Src SH2 domain. To establish the extent of the FAK binding motif, the binding affinity of the SH2 domain for phosphorylated and unphosphorylated FAK-derived peptides of increasing length was determined and compared with that of the internal Src SH2 binding site. It is shown that the FAK peptides have higher affinity than the internal binding site and that seven negative residues adjacent to the core SH2 binding motif increase the binding constant 30-fold. A rigid spin-label incorporated in the FAK peptides was used to establish on the basis of paramagnetic relaxation enhancement whether the peptide-protein complex is well defined. A large spread of the paramagnetic effects on the surface of the SH2 domain suggests that the peptide-protein complex exhibits dynamics, despite the high affinity of the peptide. The strong electrostatic interaction between the positive side of the SH2 domain and the negative peptide results in a high affinity but may also favor a dynamic interaction. Copyright © 2012 Wiley Periodicals, Inc.

Technical support to the Solvent Refined Coal (SRC) demonstration projects: assessment of current research and development

Energy Technology Data Exchange (ETDEWEB)

Edwards, M.S.; Rodgers, B.R.; Brown, C.H.; Carlson, P.K.; Gambill, W.R.; Gilliam, T.M.; Holmes, J.M.; Krishnan, R.P.; Parsly, L.F.

1980-12-01

A program to demonstrate Solvent Refined Coal (SRC) technology has been initiated by the US Department of Energy (DOE) in partnership with two industrial groups. Project management responsibility has been assigned to the Oak Ridge Operations Office (ORO) of DOE. ORO requested that the Oak Ridge National Laboratory assess current research and development (R and D) activities and develop recommendations for those activities that might contribute to successful completion of the SRC demonstration plant projects. The objectives of this final report are to discuss in detail the problem areas in SRC; to discuss the current and planned R and D investigations relevant to the problems identified; and to suggest appropriate R and D activities in support of designs for the SRC demonstration plants. Four types of R and D activities are suggested: continuation of present and planned activities; coordination of activities and results, present and proposed; extension/redirection of activities not involving major equipment purchase or modifications; and new activities. Important examples of the first type of activity include continuation of fired heater, slurry rheology, and slurry mixing studies at Ft. Lewis. Among the second type of activity, coordination of data acquisition and interpretation is recommended in the areas of heat transfer, vapor/liquid equilibria, and physical properties. Principal examples of recommendations for extension/redirection include screening studies at laboratory scale on the use of carbonaceous precoat (e.g., anthracite) infiltration, and 15- to 30-day continuous tests of the Texaco gasifier at the Texaco Montebello facility (using SRC residues).

Src controls castration recurrence of CWR22 prostate cancer xenografts

International Nuclear Information System (INIS)

Su, Bing; Gillard, Bryan; Gao, Lingqiu; Eng, Kevin H; Gelman, Irwin H

2013-01-01

Recurrence of prostate cancer (CaP) after androgen-deprivation therapy continues to have the greatest impact on patient survival. Castration-recurrent (CR)-CaP is likely driven by the activation of androgen receptor (AR) through multiple mechanisms including induction of AR coregulators, AR mutants or splice variants, and AR posttranslational modification such as phosphorylation by Src-family and Ack1 tyrosine kinases. Here, we address whether Src is required for the CR growth of human CWR22 CaP xenografts. The shRNA-mediated Src knockdown or treatment with the Src inhibitors, dasatinib or KXO1, reduced CaP recurrence over controls and increased time-to-recurrence following castration. Moreover, CR-CaP [Src-shRNA] tumors that recurred had similar Src protein and activation levels as those of parental cells, strengthening the notion that Src activity is required for progression to CR-CaP. In contrast, the ability of dasatinib or KXO1 to inhibit Src kinase activity in vitro did not correlate with their ability to inhibit serum-driven in vitro proliferation of CR and androgen-dependent stable cell lines derived from CWR22 tumors (CWR22Rv1 and CWR22PC, respectively), suggesting that the in vitro proliferation of these CaP lines is Src independent. Taken together, these findings strongly suggest that Src is a potent and specific therapeutic target for CR-CaP progression

Electrostatic effects in the folding of the SH3 domain of the c-Src tyrosine kinase: pH-dependence in 3D-domain swapping and amyloid formation.

Directory of Open Access Journals (Sweden)

Julio Bacarizo

Full Text Available The SH3 domain of the c-Src tyrosine kinase (c-Src-SH3 aggregates to form intertwined dimers and amyloid fibrils at mild acid pHs. In this work, we show that a single mutation of residue Gln128 of this SH3 domain has a significant effect on: (i its thermal stability; and (ii its propensity to form amyloid fibrils. The Gln128Glu mutant forms amyloid fibrils at neutral pH but not at mild acid pH, while Gln128Lys and Gln128Arg mutants do not form these aggregates under any of the conditions assayed. We have also solved the crystallographic structures of the wild-type (WT and Gln128Glu, Gln128Lys and Gln128Arg mutants from crystals obtained at different pHs. At pH 5.0, crystals belong to the hexagonal space group P6₅22 and the asymmetric unit is formed by one chain of the protomer of the c-Src-SH3 domain in an open conformation. At pH 7.0, crystals belong to the orthorhombic space group P2₁2₁2₁, with two molecules at the asymmetric unit showing the characteristic fold of the SH3 domain. Analysis of these crystallographic structures shows that the residue at position 128 is connected to Glu106 at the diverging β-turn through a cluster of water molecules. Changes in this hydrogen-bond network lead to the displacement of the c-Src-SH3 distal loop, resulting also in conformational changes of Leu100 that might be related to the binding of proline rich motifs. Our findings show that electrostatic interactions and solvation of residues close to the folding nucleation site of the c-Src-SH3 domain might play an important role during the folding reaction and the amyloid fibril formation.

Development of SRC-I product analysis. Volume 3. Documentation of procedures

Energy Technology Data Exchange (ETDEWEB)

Schweighardt, F.K.; Kingsley, I.S.; Cooper, F.E.; Kamzelski, A.Z.; Parees, D.M.

1983-09-01

This section documents the BASIC computer program written to simulate Wilsonville's GC-simulated distillation (GCSD) results at APCI-CRSD Trexlertown. The GC conditions used at APCI for the Wilsonville GCSD analysis of coal-derived liquid samples were described in the SRC-I Quarterly Technical Report, April-June 1981. The approach used to simulate the Wilsonville GCSD results is also from an SRC-I Quarterly Technical Report and is reproduced in Appendix VII-A. The BASIC computer program is described in the attached Appendix VII-B. Analysis of gases produced during coal liquefaction generates key information needed to determine product yields for material balance and process control. Gas samples from the coal process development unit (CPDU) and tubing bombs are the primary samples analyzed. A Carle gas chromatographic system was used to analyze coal liquefaction gas samples. A BASIC computer program was written to calculate the gas chromatographic peak area results into mole percent results. ICRC has employed several analytical workup procedures to determine the amount of distillate, oils, asphaltenes, preasphaltenes, and residue in SRC-I process streams. The ASE procedure was developed using Conoco's liquid column fractionation (LC/F) method as a model. In developing the ASE procedure, ICRC was able to eliminate distillation, and therefore quantify the oils fraction in one extraction step. ASE results were shown to be reproducible within +- 2 wt %, and to yield acceptable material balances. Finally, the ASE method proved to be the least affected by sample composition.

Src binds cortactin through an SH2 domain cystine-mediated linkage

Science.gov (United States)

Evans, Jason V.; Ammer, Amanda G.; Jett, John E.; Bolcato, Chris A.; Breaux, Jason C.; Martin, Karen H.; Culp, Mark V.; Gannett, Peter M.; Weed, Scott A.

2012-01-01

Summary Tyrosine-kinase-based signal transduction mediated by modular protein domains is critical for cellular function. The Src homology (SH)2 domain is an important conductor of intracellular signaling that binds to phosphorylated tyrosines on acceptor proteins, producing molecular complexes responsible for signal relay. Cortactin is a cytoskeletal protein and tyrosine kinase substrate that regulates actin-based motility through interactions with SH2-domain-containing proteins. The Src kinase SH2 domain mediates cortactin binding and tyrosine phosphorylation, but how Src interacts with cortactin is unknown. Here we demonstrate that Src binds cortactin through cystine bonding between Src C185 in the SH2 domain within the phosphotyrosine binding pocket and cortactin C112/246 in the cortactin repeats domain, independent of tyrosine phosphorylation. Interaction studies show that the presence of reducing agents ablates Src-cortactin binding, eliminates cortactin phosphorylation by Src, and prevents Src SH2 domain binding to cortactin. Tandem MS/MS sequencing demonstrates cystine bond formation between Src C185 and cortactin C112/246. Mutational studies indicate that an intact cystine binding interface is required for Src-mediated cortactin phosphorylation, cell migration, and pre-invadopodia formation. Our results identify a novel phosphotyrosine-independent binding mode between the Src SH2 domain and cortactin. Besides Src, one quarter of all SH2 domains contain cysteines at or near the analogous Src C185 position. This provides a potential alternative mechanism to tyrosine phosphorylation for cysteine-containing SH2 domains to bind cognate ligands that may be widespread in propagating signals regulating diverse cellular functions. PMID:23097045

Src binds cortactin through an SH2 domain cystine-mediated linkage.

Science.gov (United States)

Evans, Jason V; Ammer, Amanda G; Jett, John E; Bolcato, Chris A; Breaux, Jason C; Martin, Karen H; Culp, Mark V; Gannett, Peter M; Weed, Scott A

2012-12-15

Tyrosine-kinase-based signal transduction mediated by modular protein domains is critical for cellular function. The Src homology (SH)2 domain is an important conductor of intracellular signaling that binds to phosphorylated tyrosines on acceptor proteins, producing molecular complexes responsible for signal relay. Cortactin is a cytoskeletal protein and tyrosine kinase substrate that regulates actin-based motility through interactions with SH2-domain-containing proteins. The Src kinase SH2 domain mediates cortactin binding and tyrosine phosphorylation, but how Src interacts with cortactin is unknown. Here we demonstrate that Src binds cortactin through cystine bonding between Src C185 in the SH2 domain within the phosphotyrosine binding pocket and cortactin C112/246 in the cortactin repeats domain, independent of tyrosine phosphorylation. Interaction studies show that the presence of reducing agents ablates Src-cortactin binding, eliminates cortactin phosphorylation by Src, and prevents Src SH2 domain binding to cortactin. Tandem MS/MS sequencing demonstrates cystine bond formation between Src C185 and cortactin C112/246. Mutational studies indicate that an intact cystine binding interface is required for Src-mediated cortactin phosphorylation, cell migration, and pre-invadopodia formation. Our results identify a novel phosphotyrosine-independent binding mode between the Src SH2 domain and cortactin. Besides Src, one quarter of all SH2 domains contain cysteines at or near the analogous Src C185 position. This provides a potential alternative mechanism to tyrosine phosphorylation for cysteine-containing SH2 domains to bind cognate ligands that may be widespread in propagating signals regulating diverse cellular functions.

Hormonal Resistance And Metastasis ER-Coregulartor-Src Signaling Targeted Therapy

Science.gov (United States)

2011-09-01

Con Con ConE2 E2 E2 E2 MCF7 MCF7-PELP1-GFP vector Src-sh1 Src- sh2 R el at iv e lu m in es ce nc e ∗∗ α Fig. 1 Down regulation of Src kinase ...dasatinib significantly inhibited the growth of therapy resistant cells. Since PELP1, HER2, and Src kinase are commonly deregulated in breast cancers...Estrogen receptor, coregulators, nongenomic actions, Src kinase , therapy resistance, metastasis 56 vadlamudi@uthscsa.edu Table of Contents

Identification of a New Interaction Mode between the Src Homology 2 Domain of C-terminal Src Kinase (Csk) and Csk-binding Protein/Phosphoprotein Associated with Glycosphingolipid Microdomains♦

Science.gov (United States)

Tanaka, Hiroaki; Akagi, Ken-ichi; Oneyama, Chitose; Tanaka, Masakazu; Sasaki, Yuichi; Kanou, Takashi; Lee, Young-Ho; Yokogawa, Daisuke; Dobenecker, Marc-Werner; Nakagawa, Atsushi; Okada, Masato; Ikegami, Takahisa

2013-01-01

Proteins with Src homology 2 (SH2) domains play major roles in tyrosine kinase signaling. Structures of many SH2 domains have been studied, and the regions involved in their interactions with ligands have been elucidated. However, these analyses have been performed using short peptides consisting of phosphotyrosine followed by a few amino acids, which are described as the canonical recognition sites. Here, we report the solution structure of the SH2 domain of C-terminal Src kinase (Csk) in complex with a longer phosphopeptide from the Csk-binding protein (Cbp). This structure, together with biochemical experiments, revealed the existence of a novel binding region in addition to the canonical phosphotyrosine 314-binding site of Cbp. Mutational analysis of this second region in cells showed that both canonical and novel binding sites are required for tumor suppression through the Cbp-Csk interaction. Furthermore, the data indicate an allosteric connection between Cbp binding and Csk activation that arises from residues in the βB/βC loop of the SH2 domain. PMID:23548896

Comprehensive analysis of interactions between the Src-associated protein in mitosis of 68 kDa and the human Src-homology 3 proteome.

Directory of Open Access Journals (Sweden)

Benedikt Asbach

Full Text Available The protein Sam68 is involved in many cellular processes such as cell-cycle regulation, RNA metabolism, or signal transduction. Sam68 comprises a central RNA-binding domain flanked by unstructured tails containing docking sites for signalling proteins including seven proline-rich sequences (denoted P0 to P6 as potential SH3-domain binding motifs. To comprehensively assess Sam68-SH3-interactions, we applied a phage-display screening of a library containing all approx. 300 human SH3 domains. Thereby we identified five new (from intersectin 2, the osteoclast stimulating factor OSF, nephrocystin, sorting nexin 9, and CIN85 and seven already known high-confidence Sam68-ligands (mainly from the Src-kinase family, as well as several lower-affinity binders. Interaction of the high-affinity Sam68-binders was confirmed in independent assays in vitro (phage-ELISA, GST-pull-down and in vivo (FACS-based FRET-analysis with CFP- and YFP-tagged proteins. Fine-mapping analyses with peptides established P0, P3, P4, and P5 as exclusive docking-sites for SH3 domains, which showed varying preferences for these motifs. Mutational analyses identified individual residues within the proline-rich motifs being crucial for the interactions. Based on these data, we generated a Sam68-mutant incapable of interacting with SH3 domains any more, as subsequently demonstrated by FRET-analyses. In conclusion, we present a thorough characterization of Sam68's interplay with the SH3 proteome. The observed interaction between Sam68 and OSF complements the known Sam68-Src and OSF-Src interactions. Thus, we propose, that Sam68 functions as a classical scaffold protein in this context, assembling components of an osteoclast-specific signalling pathway.

Tyrosine phosphorylation of the Lyn Src homology 2 (SH2) domain modulates its binding affinity and specificity.

Science.gov (United States)

Jin, Lily L; Wybenga-Groot, Leanne E; Tong, Jiefei; Taylor, Paul; Minden, Mark D; Trudel, Suzanne; McGlade, C Jane; Moran, Michael F

2015-03-01

Src homology 2 (SH2) domains are modular protein structures that bind phosphotyrosine (pY)-containing polypeptides and regulate cellular functions through protein-protein interactions. Proteomics analysis showed that the SH2 domains of Src family kinases are themselves tyrosine phosphorylated in blood system cancers, including acute myeloid leukemia, chronic lymphocytic leukemia, and multiple myeloma. Using the Src family kinase Lyn SH2 domain as a model, we found that phosphorylation at the conserved SH2 domain residue Y(194) impacts the affinity and specificity of SH2 domain binding to pY-containing peptides and proteins. Analysis of the Lyn SH2 domain crystal structure supports a model wherein phosphorylation of Y(194) on the EF loop modulates the binding pocket that engages amino acid side chains at the pY+2/+3 position. These data indicate another level of regulation wherein SH2-mediated protein-protein interactions are modulated by SH2 kinases and phosphatases. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

Differential subcellular membrane recruitment of Src may specify its downstream signalling

International Nuclear Information System (INIS)

Diesbach, Philippe de; Medts, Thierry; Carpentier, Sarah; D'Auria, Ludovic; Van Der Smissen, Patrick; Platek, Anna; Mettlen, Marcel; Caplanusi, Adrian; Hove, Marie-France van den; Tyteca, Donatienne; Courtoy, Pierre J.

2008-01-01

Most Src family members are diacylated and constitutively associate with membrane 'lipid rafts' that coordinate signalling. Whether the monoacylated Src, frequently hyperactive in carcinomas, also localizes at 'rafts' remains controversial. Using polarized MDCK cells expressing the thermosensitive v-Src/tsLA31 variant, we here addressed how Src tyrosine-kinase activation may impact on its (i) membrane recruitment, in particular to 'lipid rafts'; (ii) subcellular localization; and (iii) signalling. The kinetics of Src-kinase thermoactivation correlated with its recruitment from the cytosol to sedimentable membranes where Src largely resisted solubilisation by non-ionic detergents at 4 deg. C and floated into sucrose density gradients like caveolin-1 and flotillin-2, i.e. 'lipid rafts'. By immunofluorescence, activated Src showed a dual localization, at apical endosomes/macropinosomes and at the apical plasma membrane. The plasma membrane Src pool did not colocalize with caveolin-1 and flotillin-2, but extensively overlapped GM1 labelling by cholera toxin. Severe (∼ 70%) cholesterol extraction with methyl-β-cyclodextrin (MβCD) did not abolish 'rafts' floatation, but strongly decreased Src association with floating 'rafts' and abolished its localization at the apical plasma membrane. Src activation independently activated first the MAP-kinase - ERK1/2 pathway, then the PI3-kinase - Akt pathway. MAP-kinase - ERK1/2 activation was insensitive to MβCD, which suppressed Akt phosphorylation and apical endocytosis induced by Src, both depending on the PI3-kinase pathway. We therefore suggest that activated Src is recruited at two membrane compartments, allowing differential signalling, first via ERK1/2 at 'non-raft' domains on endosomes, then via PI3-kinase-Akt on a distinct set of 'rafts' at the apical plasma membrane. Whether this model is applicable to c-Src remains to be examined

Increased radiosensitivity and radiation-induced apoptosis in SRC-3 knockout mice

International Nuclear Information System (INIS)

Jin Jie; Wang Yu; Xu Yang; Chen Shilei; Wang Junping; Ran Xinze; Su Yongping; Wang Jin

2014-01-01

Steroid receptor coactivator-3 (SRC-3), a multifunctional transcriptional coactivator, plays an important role in regulation of cell apoptosis in chemoresistant cancer cells. However, its role in radiation-induced apoptosis in hematopoietic cells is still unclear. In this study, we used SRC-3 knockout (SRC-3 -/- ) mice to assess the role of SRC-3 in radiation-induced hematopoietic injury in vivo. After a range of doses of irradiation, SRC-3 -/- mice exhibited lower counts of peripheral blood cells and bone marrow (BM) mononuclear cells and excessive BM depression, which resulted in a significantly higher mortality compared with wildtype mice. Moreover, BM mononuclear cells obtained from SRC-3 -/- mice showed a remarkable increase in radiation-induced apoptosis. Collectively, our data demonstrate that SRC-3 plays a role in radiation-induced apoptosis of BM hematopoietic cells. Regulation of SRC-3 might influence the radiosensitivity of hematopoietic cells, which highlights a potential therapeutic target for radiation-induced hematopoietic injury. (author)

The imaging performance of the SRC on Mars Express

Science.gov (United States)

Oberst, J.; Schwarz, G.; Behnke, T.; Hoffmann, H.; Matz, K.-D.; Flohrer, J.; Hirsch, H.; Roatsch, T.; Scholten, F.; Hauber, E.; Brinkmann, B.; Jaumann, R.; Williams, D.; Kirk, R.; Duxbury, T.; Leu, C.; Neukum, G.

2008-01-01

The Mars Express spacecraft carries the pushbroom scanner high-resolution stereo camera (HRSC) and its added imaging subsystem super resolution channel (SRC). The SRC is equipped with its own optical system and a 1024??1024 framing sensor. SRC produces snapshots with 2.3 m ground pixel size from the nominal spacecraft pericenter height of 250 km, which are typically embedded in the central part of the large HRSC scenes. The salient features of the SRC are its light-weight optics, a reliable CCD detector, and high-speed read-out electronics. The quality and effective visibility of details in the SRC images unfortunately falls short of what has been expected. In cases where thermal balance cannot be reached, artifacts, such as blurring and "ghost features" are observed in the images. In addition, images show large numbers of blemish pixels and are plagued by electronic noise. As a consequence, we have developed various image improving algorithms, which are discussed in this paper. While results are encouraging, further studies of image restoration by dedicated processing appear worthwhile. The SRC has obtained more than 6940 images at the time of writing (1 September 2007), which often show fascinating details in surface morphology. SRC images are highly useful for a variety of applications in planetary geology, for studies of the Mars atmosphere, and for astrometric observations of the Martian satellites. This paper will give a full account of the design philosophy, technical concept, calibration, operation, integration with HRSC, and performance, as well as science accomplishments of the SRC. ?? 2007 Elsevier Ltd. All rights reserved.

Interplay of Matrix Stiffness and c-SRC in Hepatic Fibrosis.

Directory of Open Access Journals (Sweden)

Jan eGörtzen

2015-12-01

Full Text Available Introduction:In liver fibrosis activation of hepatic stellate cells (HSC comprises phenotypical change into profibrotic and myofibroplastic cells with increased contraction and secretion of extracellular matrix (ECM proteins. The small GTPase RhoA orchestrates cytoskeleton formation, migration and mobility via non-receptor tyrosine-protein kinase c-SRC (cellular sarcoma in different cells. Furthermore, RhoA and its downstream effector Rho-kinase also play a crucial role in hepatic stellate cells and hepatic fibrogenesis. Matrix stiffness promotes HSC activation via cytoskeleton modulation. This study investigated the interaction of c-SRC and RhoA under different matrix stiffness conditions.Methods:Liver fibrosis was induced in rats using bile duct ligation (BDL, thioacetamide (TAA or carbon tetrachloride (CCl4 models. mRNA levels of albumin, PDGF-R, RHOA, COL1A1 and αSMA were analyzed via qRT-PCR. Western Blots using phospho-specific antibodies against p-c-SRC418 and p-c-SRC530 analyzed the levels of activating and inactivating c-SRC respectively. LX2 cells and hepatocytes were cultured on acrylamide gels of 1kPa and 12kPa or on plastic to mimic non-fibrotic, fibrotic or cirrhotic environments, then exposed to SRC-inhibitor PP2. Overexpression of RhoA was performed by transfection using RhoA-plasmids. Additionally, samples from cirrhotic patients and controls were collected at liver transplantations and tumor resections were analyzed for RhoA and c-SRC protein expression by Western Blot.Results:Transcription of albumin and RhoA was decreased, whereas transcription and activation of c-SRC was increased in hepatocytes cultured on 12kPa compared to 1kPa gels. LX2 cells cultured on 12kPa gels showed upregulation of RHOA, COL1A1 and αSMA mRNA levels. Inhibition of c-SRC by PP2 in LX2 cells led to an increase in COL1A1 and αSMA most prominently in 12kPa gels. In LX2 cells with RhoA overexpression, c-SRC inhibition by PP2 failed to improve fibrosis

Antiangiogenic and Antitumor Effects of Src Inhibition in Ovarian Carcinoma

Science.gov (United States)

Han, Liz Y.; Landen, Charles N.; Trevino, Jose G.; Halder, Jyotsnabaran; Lin, Yvonne G.; Kamat, Aparna A.; Kim, Tae-Jin; Merritt, William M.; Coleman, Robert L.; Gershenson, David M.; Shakespeare, William C.; Wang, Yihan; Sundaramoorth, Raji; Metcalf, Chester A.; Dalgarno, David C.; Sawyer, Tomi K.; Gallick, Gary E.; Sood, Anil K.

2011-01-01

Src, a nonreceptor tyrosine kinase, is a key mediator for multiple signaling pathways that regulate critical cellular functions and is often aberrantly activated in a number of solid tumors, including ovarian carcinoma. The purpose of this study was to determine the role of activated Src inhibition on tumor growth in an orthotopic murine model of ovarian carcinoma. In vitro studies on HeyA8 and SKOV3ip1 cell lines revealed that Src inhibition by the Src-selective inhibitor, AP23846, occurred within 1 hour and responded in a dose-dependent manner. Furthermore, Src inhibition enhanced the cytotoxicity of docetaxel in both chemosensitive and chemoresistant ovarian cancer cell lines, HeyA8 and HeyA8-MDR, respectively. In vivo, Src inhibition by AP23994, an orally bioavailable analogue of AP23846, significantly decreased tumor burden in HeyA8 (P = 0.02), SKOV3ip1 (P = 0.01), as well as HeyA8-MDR (P < 0.03) relative to the untreated controls. However, the greatest effect on tumor reduction was observed in combination therapy with docetaxel (P < 0.001, P = 0.002, and P = 0.01, for the above models, respectively). Proliferating cell nuclear antigen staining showed that Src inhibition alone (P = 0.02) and in combination with docetaxel (P = 0.007) significantly reduced tumor proliferation. In addition, Src inhibition alone and in combination with docetaxel significantly down-regulated tumoral production of vascular endothelial growth factor and interleukin 8, whereas combination therapy decreased the microvessel density (P = 0.02) and significantly affected vascular permeability (P < 0.05). In summary, Src inhibition with AP23994 has potent antiangiogenic effects and significantly reduces tumor burden in preclinical ovarian cancer models. Thus, Src inhibition may be an attractive therapeutic approach for patients with ovarian carcinoma. PMID:16951177

Valsartan regulates the interaction of angiotensin II type 1 receptor and endothelial nitric oxide synthase via Src/PI3K/Akt signalling.

Science.gov (United States)

Su, Kuo-Hui; Tsai, Jin-Yi; Kou, Yu Ru; Chiang, An-Na; Hsiao, Sheng-Huang; Wu, Yuh-Lin; Hou, Hsin-Han; Pan, Ching-Chian; Shyue, Song-Kun; Lee, Tzong-Shyuan

2009-06-01

Valsartan, a selective angiotensin II type 1 receptor (AT1R) blocker, has beneficial effects in the cardiovascular system in part by its increase of nitric oxide (NO) bioavailability, yet the mechanisms are unclear. We investigated the molecular mechanisms underlying this effect in endothelial cells (ECs). NO production was examined by Griess reagent assay, DAF-2 DA fluorescence staining and cGMP ELISA kits. Protein interaction was determined by western blotting and immunoprecipitation. Treating bovine or human aortic ECs with valsartan increased NO production, as evidenced by elevated level of stable NO metabolites and intracellular cGMP. Valsartan increased the phosphorylation but not the protein level of endothelial NO synthase (eNOS). Inhibition of phosphoinositide-3 kinase (PI3K)/Akt and Src pathways by specific inhibitors suppressed valsartan-induced NO release. In addition, valsartan increased the tyrosine residue phosphorylation of AT1R, which was attenuated by inhibition of Src but not PI3K activities. Valsartan also suppressed the interaction of eNOS and AT1R, which was blocked by Src or PI3K inhibition. Valsartan-induced NO production in ECs is mediated through Src/PI3K/Akt-dependent phosphorylation of eNOS. Valsartan-induced AT1R phosphorylation depends on Src but not PI3K, whereas valsartan-induced suppression of AT1R-eNOS interaction depends on Src/PI3K/Akt signalling. These results indicate a novel vasoprotective mechanism of valsartan in upregulating NO production in ECs.

Novel Role of Src in Priming Pyk2 Phosphorylation.

Directory of Open Access Journals (Sweden)

Ming Zhao

Full Text Available Proline-rich tyrosine kinase 2 (Pyk2 is a member of the focal adhesion kinase (FAK family of non-receptor tyrosine kinases and plays an important role in diverse cellular events downstream of the integrin-family of receptors, including cell migration, proliferation and survival. Here, we have identified a novel role for Src kinase in priming Pyk2 phosphorylation and subsequent activation upon cell attachment on the integrin-ligand fibronectin. By using complementary methods, we show that Src activity is indispensable for the initial Pyk2 phosphorylation on the Y402 site observed in response to cell attachment. In contrast, the initial fibronectin-induced autophosphorylation of FAK in the homologous Y397 site occurs in a Src-independent manner. We demonstrate that the SH2-domain of Src is required for Src binding to Pyk2 and for Pyk2 phosphorylation at sites Y402 and Y579. Moreover, Y402 phosphorylation is a prerequisite for the subsequent Y579 phosphorylation. While this initial phosphorylation of Pyk2 by Src is independent of Pyk2 kinase activity, subsequent autophosphorylation of Pyk2 in trans is required for full Pyk2 phosphorylation and activation. Collectively, our studies reveal a novel function of Src in priming Pyk2 (but not FAK phosphorylation and subsequent activation downstream of integrins, and shed light on the signaling events that regulate the function of Pyk2.

Elevated c-Src and c-Yes expression in malignant skin cancers

Directory of Open Access Journals (Sweden)

Lee Jang

2010-08-01

Full Text Available Abstracts Background Src family kinases (SFKs play an important role in cancer proliferation, survival, motility, invasiveness, metastasis, and angiogenesis. Among the SFKs, c-Src and c-Yes are particularly over-expressed or hyper-activated in many human epithelial cancers. However, only a few studies have attempted to define the expression and role of c-Src and c-Yes in cutaneous carcinomas. Objectives To investigate the expression of c-Src and c-Yes in cutaneous carcinomas to include malignant melanoma (MM, squamous cell carcinoma (SCC and basal cell carcinoma (BCC. Methods We examined 6 normal skin tissues and 18 malignant skin tumor tissues using western blotting for the expression of c-Src and c-Yes. In another set, 16 specimens of MM, 16 SCCs and 16 BCCs were analyzed for the expression of c-Src and c-Yes using immunohistochemical staining. Results Western blotting showed that c-Src was expressed in all malignant skin tumors, but not in normal skin, while c-Yes was expressed in MM and SCC, but not in BCC and normal skin. Immunohistochemical staining results of c-Src and c-Yes in MM, SCC, and BCC mirrored those of the western blot analysis. Conclusions c-Src, rather than c-Yes, plays a key role in the proliferation and progression of malignant skin cancers.

Study on Progressive Collapse Behavior of SRC Column-Steel Beam Hybrid Frame Based on Pushdown Analysis

Directory of Open Access Journals (Sweden)

Liusheng Chu

2017-01-01

Full Text Available To investigate the progressive collapse behavior of Steel Reinforced Concrete (SRC column-steel beam hybrid frame after the failure of key structural elements, a PQ-Fiber model for an 8-storey structure is established in ABAQUS program. Nonlinear dynamic and static pushdown analysis are carried out after the failure and removal of the bottom-middle and bottom-corner columns. Numerical results of both methods agree well with each other. Results show that SRC column-steel frame has good resistance to progressive collapse under dynamic instantaneous load. After sudden removal of a bottom middle column, the development of structural collapse exhibits two mechanisms, the beam mechanism and the catenary mechanism. When the structure is within small deformation range, the collapse resistance of the residual frame is provided by the beam bending moment capacity, which is beam mechanism. For large deformation situation, the collapse resistance is mainly provided by the beam tensile strength, which is catenary mechanism. However, with the removal of a bottom corner column, the residual structure only undergoes the beam mechanism even for large deformations. For future practical applications, the influence of the steel ratio, steel section size, and the vertical position of the removed key components are investigated through a detailed parametric study.

EG-1 interacts with c-Src and activates its signaling pathway.

Science.gov (United States)

Lu, Ming; Zhang, Liping; Sartippour, Maryam R; Norris, Andrew J; Brooks, Mai N

2006-10-01

EG-1 is significantly elevated in breast, colorectal, and prostate cancers. Overexpression of EG-1 stimulates cellular proliferation, and targeted inhibition blocks mouse xenograft tumor growth. To further clarify the function of EG-1, we investigated its role in c-Src activation. We observed that EG-1 overexpression results in activation of c-Src, but found no evidence that EG-1 is a direct Src substrate. EG-1 also binds to other members of the Src family. Furthermore, EG-1 shows interaction with multiple other SH3- and WW-containing molecules involved in various signaling pathways. These observations suggest that EG-1 may be involved in signaling pathways including c-Src activation.

4-Hydroxynonenal activates Src through a non-canonical pathway that involves EGFR/PTP1B

Science.gov (United States)

Zhang, Hongqiao; Forman, Henry Jay

2015-01-01

Src, a non-receptor protein tyrosine kinase involved in many biological processes, can be activated through both redox-dependent and independent mechanisms. 4-Hydroxy-2-nonenal (HNE) is a lipid peroxidation product that is increased in pathophysiological conditions associated with Src activation. This study examined how HNE activates human c-Src. In the canonical pathway Src activation is initiated by dephosphorylation of pTyr530 followed by conformational change that causes Src auto-phosphorylation at Tyr419 and its activation. HNE increased Src activation in both dose- and time-dependent manner, while it also increased Src phosphorylation at Tyr530 (pTyr530 Src), suggesting that HNE activated Src via a non-canonical mechanism. Protein tyrosine phosphatase 1B inhibitor (539741), at concentrations that increased basal pTyr530 Src, also increased basal Src activity and significantly reduced HNE-mediated Src activation. The EGFR inhibitor, AG1478, and EGFR silencing, abrogated HNE-mediated EGFR activation and inhibited basal and HNE-induced Src activity. In addition, AG1478 also eliminated the increase of basal Src activation by a PTP1B inhibitor. Taken together these data suggest that HNE can activate Src partly through a non-canonical pathway involving activation of EGFR and inhibition of PTP1B. PMID:26453921

c-Src activity is differentially required by cancer cell motility modes.

Science.gov (United States)

Logue, Jeremy S; Cartagena-Rivera, Alexander X; Chadwick, Richard S

2018-04-01

Cancer cell migration requires that cells respond and adapt to their surroundings. In the absence of extracellular matrix cues, cancer cells will undergo a mesenchymal to ameboid transition, whereas a highly confining space will trigger a switch to "leader bleb-based" migration. To identify oncogenic signaling pathways mediating these transitions, we undertook a targeted screen using clinically useful inhibitors. Elevated Src activity was found to change actin and focal adhesion dynamics, whereas inhibiting Src triggered focal adhesion disassembly and blebbing. On non-adherent substrates and in collagen matrices, amoeboid-like, blebbing cells having high Src activity formed protrusions of the plasma membrane. To evaluate the role of Src in confined cells, we use a novel approach that places cells under a slab of polydimethylsiloxane (PDMS), which is held at a defined height. Using this method, we find that leader bleb-based migration is resistant to Src inhibition. High Src activity was found to markedly change the architecture of cortical actomyosin, reduce cell mechanical properties, and the percentage of cells that undergo leader bleb-based migration. Thus, Src is a signal transducer that can potently influence transitions between migration modes with implications for the rational development of metastasis inhibitors.

SRC: marker or actor in prostate cancer aggressiveness.

Science.gov (United States)

Vlaeminck-Guillem, Virginie; Gillet, Germain; Rimokh, Ruth

2014-01-01

A key question for urologic practitioners is whether an apparently organ-confined prostate cancer (PCa) is actually aggressive or not. The dilemma is to specifically identify among all prostate tumors the very aggressive high-grade cancers that will become life-threatening by developing extra-prostatic invasion and metastatic potential and the indolent cancers that will never modify a patient's life expectancy. A choice must be made between several therapeutic options to achieve the optimal personalized management of the disease that causes as little harm as possible to patients. Reliable clinical, biological, or pathological markers that would enable distinctions to be made between aggressive and indolent PCas in routine practice at the time of initial diagnosis are still lacking. The molecular mechanisms that explain why a PCa is aggressive or not are also poorly understood. Among the potential markers and/or actors in PCa aggressiveness, Src and other members of the Src kinase family, are valuable candidates. Activation of Src-dependent intracellular pathways is frequently observed in PCa. Indeed, Src is at the cross-roads of several pathways [including androgen receptor (AR), TGFbeta, Bcl-2, Akt/PTEN or MAPK, and ERK …], and is now known to influence some of the cellular and tissular events that accompany tumor progression: cell proliferation, cell motility, invasion, epithelial-to-mesenchymal transition, resistance to apoptosis, angiogenesis, neuroendocrine differentiation, and metastatic spread. Recent work even suggests that Src could also play a part in PCa initiation in coordination with the AR. The aim of this review is to gather data that explore the links between the Src kinase family and PCa progression and aggressiveness.

Src: marker or actor of prostate cancer aggressiveness

Directory of Open Access Journals (Sweden)

Virginie eVlaeminck-Guillem

2014-08-01

Full Text Available A key question for urologic practitioners is whether an apparently organ-confined prostate cancer is actually aggressive or not. The dilemma is to specifically identify among all prostate tumors the very aggressive high-grade cancers that will become life-threatening by developing extra-prostatic invasion and metastatic potential and the indolent cancers that will never modify a patient’s life expectancy. A choice must be made between several therapeutic options to achieve the optimal personalized management of the disease that causes as little harm as possible to patients. Reliable clinical, biological or pathological markers that would enable distinctions to be made between aggressive and indolen prostate cancers in routine practice at the time of initial diagnosis are still lacking. The molecular mechanisms that explain why a prostate cancer is aggressive or not are also poorly understood. Among the potential markers and/or actors in prostate cancer aggressiveness, Src and other members of the Src kinase family, are valuable candidates. Activation of Src-dependent intracellular pathways is frequently observed in prostate cancer. Indeed, Src is at the cross-roads of several pathways (including androgen receptor, TGFbeta, Bcl-2, Akt/PTEN or MAPK and ERK …, and is now known to influence some of the cellular and tissular events that accompany tumor progression: cell proliferation, cell motility, invasion, epithelial-to-mesenchymal transition, resistance to apoptosis, angiogenesis, neuroendocrine differentiation, and metastatic spread. Recent work even suggests that Src could also play a part in prostate cancer initiation in coordination with the androgen receptor. The aim of this review is to gather data that explores the links between the Src kinase family and prostate cancer progression and aggressiveness.

Roles of Raft-Anchored Adaptor Cbp/PAG1 in Spatial Regulation of c-Src Kinase

Science.gov (United States)

Oneyama, Chitose; Suzuki, Takashi; Okada, Masato

2014-01-01

The tyrosine kinase c-Src is upregulated in numerous human cancers, implying a role for c-Src in cancer progression. Previously, we have shown that sequestration of activated c-Src into lipid rafts via a transmembrane adaptor, Cbp/PAG1, efficiently suppresses c-Src-induced cell transformation in Csk-deficient cells, suggesting that the transforming activity of c-Src is spatially regulated via Cbp in lipid rafts. To dissect the molecular mechanisms of the Cbp-mediated regulation of c-Src, a combined analysis was performed that included mathematical modeling and in vitro experiments in a c-Src- or Cbp-inducible system. c-Src activity was first determined as a function of c-Src or Cbp levels, using focal adhesion kinase (FAK) as a crucial c-Src substrate. Based on these experimental data, two mathematical models were constructed, the sequestration model and the ternary model. The computational analysis showed that both models supported our proposal that raft localization of Cbp is crucial for the suppression of c-Src function, but the ternary model, which includes a ternary complex consisting of Cbp, c-Src, and FAK, also predicted that c-Src function is dependent on the lipid-raft volume. Experimental analysis revealed that c-Src activity is elevated when lipid rafts are disrupted and the ternary complex forms in non-raft membranes, indicating that the ternary model accurately represents the system. Moreover, the ternary model predicted that, if Cbp enhances the interaction between c-Src and FAK, Cbp could promote c-Src function when lipid rafts are disrupted. These findings underscore the crucial role of lipid rafts in the Cbp-mediated negative regulation of c-Src-transforming activity, and explain the positive role of Cbp in c-Src regulation under particular conditions where lipid rafts are perturbed. PMID:24675741

Densification of chipper harvested SRC using on-farm machinery

Energy Technology Data Exchange (ETDEWEB)

Paulson, M.

2003-07-01

This report gives details of a project to density wood chips using on-farm machinery in order to avoid problems encountered in bulk handling and storage of low density short rotation cultivation (SRC) wood chips - especially as some customers can only accept baled material. Trials using different lengths of chips produced by a standard SRC harvester are described, and the failure to produce acceptable bales is reported. The potential cost of modifying equipment is deemed to make the baling of SRC chips using standard farm machinery currently not viable.

Specific Inhibition of SRC Kinase Impairs Malignant Glioma Growth In Vitro and In Vivo

Directory of Open Access Journals (Sweden)

Hanna Stedt

2012-01-01

Full Text Available Malignant glioma is a severe cancer with a poor prognosis. Local occurrence and rare metastases of malignant glioma make it a suitable target for gene therapy. Several studies have demonstrated the importance of Src kinase in different cancers. However, these studies have focused mainly on Src-deficient mice or pharmacological inhibitors of Src. In this study we have used Src small hairpin RNAs (shRNAs in a lentiviral backbone to mimic a long-term stable treatment and determined the role of Src in tumor tissues. Efficacy of Src shRNAs was confirmed in vitro demonstrating up to 90% target gene inhibition. In a mouse malignant glioma model, Src shRNA tumors were almost 50-fold smaller in comparison to control tumors and had significantly reduced vascularity. In a syngenic rat intracranial glioma model, Src shRNA-transduced tumors were smaller and these rats had a survival benefit over the control rats. In vivo treatment was enhanced by chemotherapy and histone deacetylase inhibition. Our results emphasise the importance of Src in tumorigenesis and demonstrate that it can be efficiently inhibited in vitro and in vivo in two independent malignant glioma models. In conclusion, Src is a potential target for RNA interference-mediated treatment of malignant glioma.

A Protein Scaffold Coordinates SRC-Mediated JNK Activation in Response to Metabolic Stress.

Science.gov (United States)

Kant, Shashi; Standen, Claire L; Morel, Caroline; Jung, Dae Young; Kim, Jason K; Swat, Wojciech; Flavell, Richard A; Davis, Roger J

2017-09-19

Obesity is a major risk factor for the development of metabolic syndrome and type 2 diabetes. How obesity contributes to metabolic syndrome is unclear. Free fatty acid (FFA) activation of a non-receptor tyrosine kinase (SRC)-dependent cJun NH 2 -terminal kinase (JNK) signaling pathway is implicated in this process. However, the mechanism that mediates SRC-dependent JNK activation is unclear. Here, we identify a role for the scaffold protein JIP1 in SRC-dependent JNK activation. SRC phosphorylation of JIP1 creates phosphotyrosine interaction motifs that bind the SH2 domains of SRC and the guanine nucleotide exchange factor VAV. These interactions are required for SRC-induced activation of VAV and the subsequent engagement of a JIP1-tethered JNK signaling module. The JIP1 scaffold protein, therefore, plays a dual role in FFA signaling by coordinating upstream SRC functions together with downstream effector signaling by the JNK pathway. Copyright © 2017 The Author(s). Published by Elsevier Inc. All rights reserved.

A Protein Scaffold Coordinates SRC-Mediated JNK Activation in Response to Metabolic Stress

Directory of Open Access Journals (Sweden)

Shashi Kant

2017-09-01

Full Text Available Obesity is a major risk factor for the development of metabolic syndrome and type 2 diabetes. How obesity contributes to metabolic syndrome is unclear. Free fatty acid (FFA activation of a non-receptor tyrosine kinase (SRC-dependent cJun NH2-terminal kinase (JNK signaling pathway is implicated in this process. However, the mechanism that mediates SRC-dependent JNK activation is unclear. Here, we identify a role for the scaffold protein JIP1 in SRC-dependent JNK activation. SRC phosphorylation of JIP1 creates phosphotyrosine interaction motifs that bind the SH2 domains of SRC and the guanine nucleotide exchange factor VAV. These interactions are required for SRC-induced activation of VAV and the subsequent engagement of a JIP1-tethered JNK signaling module. The JIP1 scaffold protein, therefore, plays a dual role in FFA signaling by coordinating upstream SRC functions together with downstream effector signaling by the JNK pathway.

Activation of Src kinase by protein-tyrosine phosphatase-PEST in osteoclasts: comparative analysis of the effects of bisphosphonate and protein-tyrosine phosphatase inhibitor on Src activation in vitro.

Science.gov (United States)

Chellaiah, Meenakshi A; Schaller, Michael D

2009-08-01

PTP-PEST is involved in the regulation of sealing ring formation in osteoclasts. In this article, we have shown a regulatory role for PTP-PEST on dephosphorylation of c-Src at Y527 and phosphorylation at Y418 in the catalytic site. Activation of Src in osteoclasts by over-expression of PTP-PEST resulted in the phosphorylation of cortactin at Y421 and WASP at Y294. Also enhanced as a result, is the interaction of Src, cortactin, and Arp2 with WASP. Moreover, the number of osteoclasts displaying sealing ring and bone resorbing activity was increased in response to PTP-PEST over-expression as compared with control osteoclasts. Cells expressing constitutively active-Src (527YDeltaF) simulate the effects mediated by PTP-PEST. Treatment of osteoclasts with a bisphosphonate alendronate or a potent PTP inhibitor PAO decreased the activity and phosphorylation of Src at Y418 due to reduced dephosphorylation state at Y527. Therefore, Src-mediated phosphorylation of cortactin and WASP as well as the formation of WASP.cortactin.Arp2 complex and sealing ring were reduced in these osteoclasts. Similar effects were observed in osteoclasts treated with an Src inhibitor PP2. We have shown that bisphosphonates could modulate the function of osteoclasts by inhibiting downstream signaling mediated by PTP-PEST/Src, in addition to its effect on the inhibition of the post-translational modification of small GTP-binding proteins such as Rab, Rho, and Rac as shown by others. The promising effects of the inhibitors PP2 and PAO on osteoclast function suggest a therapeutic approach for patients with bone metastases and osteoporosis as an alternative to bisphosphonates.

The role of Src kinase in the biology and pathogenesis of Acanthamoeba castellanii

Directory of Open Access Journals (Sweden)

Siddiqui Ruqaiyyah

2012-06-01

Full Text Available Abstract Background Acanthamoeba species are the causative agents of fatal granulomatous encephalitis in humans. Haematogenous spread is thought to be a primary step, followed by blood–brain barrier penetration, in the transmission of Acanthmaoeba into the central nervous system, but the associated molecular mechanisms remain unclear. Here, we evaluated the role of Src, a non-receptor protein tyrosine kinase in the biology and pathogenesis of Acanthamoeba. Methods Amoebistatic and amoebicidal assays were performed by incubating amoeba in the presence of Src kinase-selective inhibitor, PP2 (4-amino-5-(4-chlorophenyl-7-(t-butylpyrazolo[3,4-d]pyrimidine and its inactive analog, PP3 (4-amino-7-phenylpyrazolo[3,4-d]pyrimidine. Using this inhibitor, the role of Src kinase in A. castellanii interactions with Escherichia coli was determined. Zymographic assays were performed to study effects of Src kinase on extracellular proteolytic activities of A. castellanii. The human brain microvascular endothelial cells were used to determine the effects of Src kinase on A. castellanii adhesion to and cytotoxicity of host cells. Results Inhibition of Src kinase using a specific inhibitor, PP2 (4-amino-5-(4 chlorophenyl-7-(t-butylpyrazolo [3,4-d] pyrimidine but not its inactive analog, PP3 (4-amino-7-phenylpyrazolo[3,4-d] pyrimidine, had detrimental effects on the growth of A. castellanii (keratitis isolate, belonging to the T4 genotype. Interestingly, inhibition of Src kinase hampered the phagocytic ability of A. castellanii, as measured by the uptake of non-invasive bacteria, but, on the contrary, invasion by pathogenic bacteria was enhanced. Zymographic assays revealed that inhibition of Src kinases reduced extracellular protease activities of A. castellanii. Src kinase inhibition had no significant effect on A. castellanii binding to and cytotoxicity of primary human brain microvascular endothelial cells, which constitute the blood–brain barrier

Mechanism of c-Src Synergy with the EGFR in Breast Cancer

National Research Council Canada - National Science Library

Tice, David

1997-01-01

.... To gain further insights into the mechanism of c-Src synergy with the EGFR, stable cell lines containing various c-Src mutants and overexpressed wt EGFR were generated and examined for tumorigenic...

Reciprocally coupled residues crucial for protein kinase Pak2 activity calculated by statistical coupling analysis.

Directory of Open Access Journals (Sweden)

Yuan-Hao Hsu

2010-03-01

Full Text Available Regulation of Pak2 activity involves at least two mechanisms: (i phosphorylation of the conserved Thr(402 in the activation loop and (ii interaction of the autoinhibitory domain (AID with the catalytic domain. We collected 482 human protein kinase sequences from the kinome database and globally mapped the evolutionary interactions of the residues in the catalytic domain with Thr(402 by sequence-based statistical coupling analysis (SCA. Perturbation of Thr(402 (34.6% suggests a communication pathway between Thr(402 in the activation loop, and Phe(387 (DeltaDeltaE(387F,402T = 2.80 in the magnesium positioning loop, Trp(427 (DeltaDeltaE(427W,402T = 3.12 in the F-helix, and Val(404 (DeltaDeltaE(404V,402T = 4.43 and Gly(405 (DeltaDeltaE(405G,402T = 2.95 in the peptide positioning loop. When compared to the cAMP-dependent protein kinase (PKA and Src, the perturbation pattern of threonine phosphorylation in the activation loop of Pak2 is similar to that of PKA, and different from the tyrosine phosphorylation pattern of Src. Reciprocal coupling analysis by SCA showed the residues perturbed by Thr(402 and the reciprocal coupling pairs formed a network centered at Trp(427 in the F-helix. Nine pairs of reciprocal coupling residues crucial for enzymatic activity and structural stabilization were identified. Pak2, PKA and Src share four pairs. Reciprocal coupling residues exposed to the solvent line up as an activation groove. This is the inhibitor (PKI binding region in PKA and the activation groove for Pak2. This indicates these evolutionary conserved residues are crucial for the catalytic activity of PKA and Pak2.

The dual kinase complex FAK-Src as a promising therapeutic target in cancer

Science.gov (United States)

Bolós, Victoria; Gasent, Joan Manuel; López-Tarruella, Sara; Grande, Enrique

2010-01-01

Focal adhesion kinase (FAK) and steroid receptor coactivator (Src) are intracellular (nonreceptor) tyrosine kinases that physically and functionally interact to promote a variety of cellular responses. Plenty of reports have already suggested an additional central role for this complex in cancer through its ability to promote proliferation and anoikis resistance in tumor cells. An important role for the FAK/Src complex in tumor angiogenesis has also been established. Furthermore, FAK and Src have been associated with solid tumor metastasis through their ability to promote the epithelial mesenchymal transition. In fact, a strong correlation between increased FAK/Src expression/phosphorylation and the invasive phenotype in human tumors has been found. Additionally, an association for FAK/Src with resistances to the current anticancer therapies has already been established. Currently, novel anticancer agents that target FAK or Src are under development in a broad variety of solid tumors. In this article we will review the normal cellular functions of the FAK/Src complex as an effector of integrin and/or tyrosine kinase receptor signaling. We will also collect data about their role in cancer and we will summarize the most recent data from the FAK and Src inhibitors under clinical and preclinical development. Furthermore, the association of both these proteins with chemotherapy and hormonal therapy resistances, as a rationale for new combined therapeutic approaches with these novel agents, to abrogate treatment associated resistances, will also be reviewed. PMID:20616959

Integrated crop management of SRC plantations to maximise crop value, wildlife benefits and other added value opportunities

Energy Technology Data Exchange (ETDEWEB)

Sage, R; Tucker, K

1998-07-01

This report summaries the results of a study aiming to develop an integrated approach to pest management (IPM) for the short rotation cultivation (SRC) of willows and poplars. Details are given of crop and site characteristics, non-destructive assessment of SRC biomass, the quantification of crop shadiness, and the effects of wind exposure on crop growth. The section on invertebrates covers invertebrates colonising UK SRC plantations, invertebrates which are or can become pests, natural control agents of SRC pests, the abundance and distribution of chrysomelids between sites, preferences exhibited by chrysomelids for different varieties, overwintering and dispersal of chrysomelids into SRC, and IPM of insects. The section on vertebrate fauna addresses birds in winter, the breeding birds of SRC, gamebird use of SRC, and mammals and other vertebrates of SRC. A section on ground flora deals with changes in ground flora with time, ground flora introductions, the effects of weeds on the growth of SRC, and an overview of integrated crop management in SRC plantations.

The dual kinase complex FAK-Src as a promising therapeutic target in cancer

Directory of Open Access Journals (Sweden)

Victoria Bolós

2010-06-01

Full Text Available Victoria Bolós1,*, Joan Manuel Gasent2,*, Sara López-Tarruella3, Enrique Grande1,#1Pfizer Oncology, Madrid, Spain; 2Hospital Gral. Universitario Marina Alta, Oncology Department, Denia Alicante, 3,#Hospital Clínico San Carlos, Oncology Department, ∗These authors contributed equally to this work, #Center affiliated to the Red Temática de Investigación Cooperativa (RD06/0020/0021. Instituto de Salud Carlos III (ISCIII, Spanish Ministry of Science and InnovationAbstract: Focal adhesion kinase (FAK and steroid receptor coactivator (Src are intracellular (nonreceptor tyrosine kinases that physically and functionally interact to promote a variety of cellular responses. Plenty of reports have already suggested an additional central role for this complex in cancer through its ability to promote proliferation and anoikis resistance in tumor cells. An important role for the FAK/Src complex in tumor angiogenesis has also been established. Furthermore, FAK and Src have been associated with solid tumor metastasis through their ability to promote the epithelial mesenchymal transition. In fact, a strong correlation between increased FAK/Src expression/phosphorylation and the invasive phenotype in human tumors has been found. Additionally, an association for FAK/Src with resistances to the current anticancer therapies has already been established. Currently, novel anticancer agents that target FAK or Src are under development in a broad variety of solid tumors. In this article we will review the normal cellular functions of the FAK/Src complex as an effector of integrin and/or tyrosine kinase receptor signaling. We will also collect data about their role in cancer and we will summarize the most recent data from the FAK and Src inhibitors under clinical and preclinical development. Furthermore, the association of both these proteins with chemotherapy and hormonal therapy resistances, as a rationale for new combined therapeutic approaches with these novel

76 FR 57763 - Alaska Region's Subsistence Resource Commission (SRC) Program

Science.gov (United States)

2011-09-16

... Hunting Plan Recommendation 10-01 12. New Business 13. Public and other Agency Comments 14. SRC Work... the meeting dates and location are changed, a notice will be published in local newspapers and announced on local radio stations prior to the meeting date. SRC meeting locations and dates may need to be...

Phosphopeptide occupancy and photoaffinity cross-linking of the v-Src SH2 domain attenuates tyrosine kinase activity.

Science.gov (United States)

Garcia, P; Shoelson, S E; Drew, J S; Miller, W T

1994-12-02

Phosphorylation of c-Src at carboxyl-terminal Tyr-527 suppresses tyrosine kinase activity and transforming potential, presumably by facilitating the intramolecular interaction of the C terminus of Src with its SH2 domain. In addition, it has been shown previously that occupancy of the c-Src SH2 domain with a phosphopeptide stimulates c-Src kinase catalytic activity. We have performed analogous studies with v-Src, the transforming protein from Rous sarcoma virus, which has extensive homology with c-Src. v-Src lacks an autoregulatory phosphorylation site, and its kinase domain is constitutively active. Phosphopeptides corresponding to the sequences surrounding c-Src Tyr-527 and a Tyr-Glu-Glu-Ile motif from the hamster polyoma virus middle T antigen inhibit tyrosine kinase activity of baculovirus-expressed v-Src 2- and 4-fold, respectively. To determine the mechanism of this regulation, the Tyr-527 phosphopeptide was substituted with the photoactive amino acid p-benzoylphenylalanine at the adjacent positions (N- and C-terminal) to phosphotyrosine. These peptides photoinactivate the v-Src tyrosine kinase 5-fold in a time- and concentration-dependent manner. Furthermore, the peptides cross-link an isolated Src SH2 domain with similar rates and specificity. These data indicate that occupancy of the v-Src SH2 domain induces a conformational change that is transmitted to the kinase domain and attenuates tyrosine kinase activity.

Inhibition of SRC-3 enhances sensitivity of human cancer cells to histone deacetylase inhibitors

Energy Technology Data Exchange (ETDEWEB)

Zou, Zhengzhi, E-mail: zouzhengzhi@m.scnu.edu.cn [MOE Key Laboratory of Laser Life Science and Institute of Laser Life Science, College of Biophotonics, South China Normal University, Guangzhou 510000 (China); Luo, Xiaoyong [Department of Oncology, The Affiliated Luoyang Central Hospital of Zhengzhou University, Luoyang 471000 (China); Nie, Peipei [KingMed Diagnostics and KingMed School of Laboratory Medicine, Guangzhou Medical University, Guangzhou 510000 (China); Wu, Baoyan; Zhang, Tao; Wei, Yanchun [MOE Key Laboratory of Laser Life Science and Institute of Laser Life Science, College of Biophotonics, South China Normal University, Guangzhou 510000 (China); Wang, Wenyi [Xiamen Cancer Center, Department of Medical Oncology, The First Affiliated Hospital of Xiamen University, Xiamen 361000 (China); Geng, Guojun; Jiang, Jie [Xiamen Cancer Center, Department of Thoracic Surgery, The First Affiliated Hospital of Xiamen University, Xiamen 361000 (China); Mi, Yanjun, E-mail: myjgj_77@163.com [Xiamen Cancer Center, Department of Medical Oncology, The First Affiliated Hospital of Xiamen University, Xiamen 361000 (China)

2016-09-09

SRC-3 is widely expressed in multiple tumor types and involved in cancer cell proliferation and apoptosis. Histone deacetylase (HDAC) inhibitors are promising antitumor drugs. However, the poor efficacy of HDAC inhibitors in solid tumors has restricted its further clinical application. Here, we reported the novel finding that depletion of SRC-3 enhanced sensitivity of breast and lung cancer cells to HDAC inhibitors (SAHA and romidepsin). In contrast, overexpression of SRC-3 decreased SAHA-induced cancer cell apoptosis. Furthermore, we found that SRC-3 inhibitor bufalin increased cancer cell apoptosis induced by HDAC inhibitors. The combination of bufalin and SAHA was particular efficient in attenuating AKT activation and reducing Bcl-2 levels. Taken together, these accumulating data might guide development of new breast and lung cancer therapies. - Highlights: • Depletion of SRC-3 enhanced sensitivity of breast and lung cancer cells to HDAC inhibitors. • Overexpression of SRC-3 enhanced cancer cell resistance to HDAC inhibitors. • SRC-3 inhibitor bufalin increased cancer cell apoptosis induced by HDAC inhibitors. • Bufalin synergized with HDAC inhibitor attenuated AKT activation and reduced Bcl-2 levels in human cancer cell.

Identification of a functional interaction between Kv4.3 channels and c-Src tyrosine kinase.

Science.gov (United States)

Gomes, Pedro; Saito, Tomoaki; Del Corsso, Cris; Alioua, Abderrahmane; Eghbali, Mansoureh; Toro, Ligia; Stefani, Enrico

2008-10-01

Voltage-gated K(+) (Kv) channels are key determinants of cardiac and neuronal excitability. A substantial body of evidence has accumulated in support of a role for Src family tyrosine kinases in the regulation of Kv channels. In this study, we examined the possibility that c-Src tyrosine kinase participates in the modulation of the transient voltage-dependent K(+) channel Kv4.3. Supporting a mechanistic link between Kv4.3 and c-Src, confocal microscopy analysis of HEK293 cells stably transfected with Kv4.3 showed high degree of co-localization of the two proteins at the plasma membrane. Our results further demonstrate an association between Kv4.3 and c-Src by co-immunoprecipitation and GST pull-down assays, this interaction being mediated by the SH2 and SH3 domains of c-Src. Furthermore, we show that Kv4.3 is tyrosine phosphorylated under basal conditions. The functional relevance of the observed interaction between Kv4.3 and c-Src was established in patch-clamp experiments, where application of the Src inhibitor PP2 caused a decrease in Kv4.3 peak current amplitude, but not the inactive structural analogue PP3. Conversely, intracellular application of recombinant c-Src kinase or the protein tyrosine phosphatase inhibitor bpV(phen) increased Kv4.3 peak current amplitude. In conclusion, our findings provide evidence that c-Src-induced Kv4.3 channel activation involves their association in a macromolecular complex and suggest a role for c-Src-Kv4.3 pathway in regulating cardiac and neuronal excitability.

Fertilization of SRC willow. II

DEFF Research Database (Denmark)

Sevel, L; Ingerslev, Morten; Nord-Larsen, Thomas

2014-01-01

impacts of different doses of mineral fertilizer, manure and sewage sludge in a commercially grown SRC willow stand. We examined macro nutrient and heavy metal leaching rates and calculated element balances to evaluate the environmental impact. Growth responses were reported in a former paper (Sevel et al...

Preliminary study of semi-refined carrageenan (SRC) as secondary gelling agent in natural rubber (NR) latex foam

Science.gov (United States)

Norhazariah, S.; Azura, A. R.; Azahari, B.; Sivakumar, R.

2017-12-01

Semi-refined carrageenan (SRC) product is considerably cheaper and easier to produce as a natural polysaccharide, which was utilized in food and other product application. However, the application in latex is limited. The aim of this work is to evaluate the SRC produced from low industrial grade seaweed (LIGS) in the latex foam application. The FTIR spectra showed the SRC produced as kappa type carrageenan with lower sulfur content compared to native LIGS. NR latex foam is produced by using the Dunlop method with some modifications. The effect of SRC loading as a secondary gelling agent in NR latex foam is investigated. The density and morphology of the NR latex foam with the addition of the SRC are analyzed. NR latex foam density increased with SRC loading and peaked at 1.8 phr SRC. The addition of SRC has induced the bigger cell size compared to the cell size of the control NR latex foam, as shown in the optical micrograph. It can be concluded that SRC LIGS could be acted as secondary gelling agent in NR latex foam.

SH2 Ligand-Like Effects of Second Cytosolic Domain of Na/K-ATPase α1 Subunit on Src Kinase.

Science.gov (United States)

Banerjee, Moumita; Duan, Qiming; Xie, Zijian

2015-01-01

Our previous studies have suggested that the α1 Na/K-ATPase interacts with Src to form a receptor complex. In vitro binding assays indicate an interaction between second cytosolic domain (CD2) of Na/K-ATPase α1 subunit and Src SH2 domain. Since SH2 domain targets Src to specific signaling complexes, we expressed CD2 as a cytosolic protein and studied whether it could act as a Src SH2 ligand in LLC-PK1 cells. Co-immunoprecipitation analyses indicated a direct binding of CD2 to Src, consistent with the in vitro binding data. Functionally, CD2 expression increased basal Src activity, suggesting a Src SH2 ligand-like property of CD2. Consistently, we found that CD2 expression attenuated several signaling pathways where Src plays an important role. For instance, although it increased surface expression of Na/K-ATPase, it decreased ouabain-induced activation of Src and ERK by blocking the formation of Na/K-ATPase/Src complex. Moreover, it also attenuated cell attachment-induced activation of Src/FAK. Consequently, CD2 delayed cell spreading, and inhibited cell proliferation. Furthermore, these effects appear to be Src-specific because CD2 expression had no effect on EGF-induced activation of EGF receptor and ERK. Hence, the new findings indicate the importance of Na/K-ATPase/Src interaction in ouabain-induced signal transduction, and support the proposition that the CD2 peptide may be utilized as a Src SH2 ligand capable of blocking Src-dependent signaling pathways via a different mechanism from a general Src kinase inhibitor.

Predicting Sensitivity of Breast Tumors to Src-Targeted Therapies through Assessment of Cas/Src/BCAR3 Activity

Science.gov (United States)

2016-10-01

GFP-BCAR3 or R171V/L744E/R748E GFP-BCAR3 and lysed in a non- denaturing buffer 24 h post transfection. Total cell protein and Cas immune complexes...assess the role of a signaling pathway comprised of the protein tyrosine kinase c-Src (Src) and two adaptor molecules, Cas and BCAR3, in promoting...1. Introduction …………………………………………………..……….4 2. Keywords………………………………………………………..…….4 3. Accomplishments………..…………………………………….……...4 4

Significance of ERa and c-Src Interaction in the Progression of Hormone Independent Breast Cancer

Science.gov (United States)

2005-12-01

defects in estrogen signaling [1]. Because of global defects in estrogen signaling observed in these c-Src deficient mice, we have recently generated...1998). Interestingly, the region of the kinase domain of ErbB-2 that correlates with c-Src association, referred to as TK2 (Segatto et al., 1991...ductive organs that are dependent on ERa in c-Src- deficient mice. We show that the loss of the c-Src tyrosine kinase correlates with defects in ductal

v-Src causes delocalization of Mklp1, Aurora B, and INCENP from the spindle midzone during cytokinesis failure

Energy Technology Data Exchange (ETDEWEB)

Soeda, Shuhei [Department of Molecular Cell Biology, Graduate School of Pharmaceutical Sciences, Chiba University, Inohana 1-8-1, Chuo-ku, Chiba 260-8675 (Japan); Nakayama, Yuji, E-mail: nakayama@mb.kyoto-phu.ac.jp [Department of Molecular Cell Biology, Graduate School of Pharmaceutical Sciences, Chiba University, Inohana 1-8-1, Chuo-ku, Chiba 260-8675 (Japan); Department of Biochemistry and Molecular Biology, Kyoto Pharmaceutical University, 5 Nakauchi-cho, Misasagi, Yamashina-ku, Kyoto 607-8414 (Japan); Honda, Takuya; Aoki, Azumi; Tamura, Naoki; Abe, Kohei; Fukumoto, Yasunori [Department of Molecular Cell Biology, Graduate School of Pharmaceutical Sciences, Chiba University, Inohana 1-8-1, Chuo-ku, Chiba 260-8675 (Japan); Yamaguchi, Naoto, E-mail: nyama@faculty.chiba-u.jp [Department of Molecular Cell Biology, Graduate School of Pharmaceutical Sciences, Chiba University, Inohana 1-8-1, Chuo-ku, Chiba 260-8675 (Japan)

2013-06-10

Src-family tyrosine kinases are aberrantly activated in cancers, and this activation is associated with malignant tumor progression. v-Src, encoded by the v-src transforming gene of the Rous sarcoma virus, is a mutant variant of the cellular proto-oncogene c-Src. Although investigations with temperature sensitive mutants of v-Src have shown that v-Src induces many oncogenic processes, the effects on cell division are unknown. Here, we show that v-Src inhibits cellular proliferation of HCT116, HeLa S3 and NIH3T3 cells. Flow cytometry analysis indicated that inducible expression of v-Src results in an accumulation of 4N cells. Time-lapse analysis revealed that binucleation is induced through the inhibition of cytokinesis, a final step of cell division. The localization of Mklp1, which is essential for cytokinesis, to the spindle midzone is inhibited in v-Src-expressing cells. Intriguingly, Aurora B, which regulates Mklp1 localization at the midzone, is delocalized from the spindle midzone and the midbody but not from the metaphase chromosomes upon v-Src expression. Mklp2, which is responsible for the relocation of Aurora B from the metaphase chromosomes to the spindle midzone, is also lost from the spindle midzone. These results suggest that v-Src inhibits cytokinesis through the delocalization of Mklp1 and Aurora B from the spindle midzone, resulting in binucleation. -- Highlights: • v-Src inhibits cell proliferation of HCT116, HeLa S3 and NIH3T3 cells. • v-Src induces binucleation together with cytokinesis failure. • v-Src causes delocalization of Mklp1, Aurora B and INCENP from the spindle midzone.

SH2 Ligand-Like Effects of Second Cytosolic Domain of Na/K-ATPase α1 Subunit on Src Kinase.

Directory of Open Access Journals (Sweden)

Moumita Banerjee

Full Text Available Our previous studies have suggested that the α1 Na/K-ATPase interacts with Src to form a receptor complex. In vitro binding assays indicate an interaction between second cytosolic domain (CD2 of Na/K-ATPase α1 subunit and Src SH2 domain. Since SH2 domain targets Src to specific signaling complexes, we expressed CD2 as a cytosolic protein and studied whether it could act as a Src SH2 ligand in LLC-PK1 cells. Co-immunoprecipitation analyses indicated a direct binding of CD2 to Src, consistent with the in vitro binding data. Functionally, CD2 expression increased basal Src activity, suggesting a Src SH2 ligand-like property of CD2. Consistently, we found that CD2 expression attenuated several signaling pathways where Src plays an important role. For instance, although it increased surface expression of Na/K-ATPase, it decreased ouabain-induced activation of Src and ERK by blocking the formation of Na/K-ATPase/Src complex. Moreover, it also attenuated cell attachment-induced activation of Src/FAK. Consequently, CD2 delayed cell spreading, and inhibited cell proliferation. Furthermore, these effects appear to be Src-specific because CD2 expression had no effect on EGF-induced activation of EGF receptor and ERK. Hence, the new findings indicate the importance of Na/K-ATPase/Src interaction in ouabain-induced signal transduction, and support the proposition that the CD2 peptide may be utilized as a Src SH2 ligand capable of blocking Src-dependent signaling pathways via a different mechanism from a general Src kinase inhibitor.

Two amino acid residues confer different binding affinities of Abelson family kinase SRC homology 2 domains for phosphorylated cortactin.

Science.gov (United States)

Gifford, Stacey M; Liu, Weizhi; Mader, Christopher C; Halo, Tiffany L; Machida, Kazuya; Boggon, Titus J; Koleske, Anthony J

2014-07-11

The closely related Abl family kinases, Arg and Abl, play important non-redundant roles in the regulation of cell morphogenesis and motility. Despite similar N-terminal sequences, Arg and Abl interact with different substrates and binding partners with varying affinities. This selectivity may be due to slight differences in amino acid sequence leading to differential interactions with target proteins. We report that the Arg Src homology (SH) 2 domain binds two specific phosphotyrosines on cortactin, a known Abl/Arg substrate, with over 10-fold higher affinity than the Abl SH2 domain. We show that this significant affinity difference is due to the substitution of arginine 161 and serine 187 in Abl to leucine 207 and threonine 233 in Arg, respectively. We constructed Abl SH2 domains with R161L and S187T mutations alone and in combination and find that these substitutions are sufficient to convert the low affinity Abl SH2 domain to a higher affinity "Arg-like" SH2 domain in binding to a phospho-cortactin peptide. We crystallized the Arg SH2 domain for structural comparison to existing crystal structures of the Abl SH2 domain. We show that these two residues are important determinants of Arg and Abl SH2 domain binding specificity. Finally, we expressed Arg containing an "Abl-like" low affinity mutant Arg SH2 domain (L207R/T233S) and find that this mutant, although properly localized to the cell periphery, does not support wild type levels of cell edge protrusion. Together, these observations indicate that these two amino acid positions confer different binding affinities and cellular functions on the distinct Abl family kinases. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

Antitumor Effect of KX-01 through Inhibiting Src Family Kinases and Mitosis.

Science.gov (United States)

Kim, Seongyeong; Min, Ahrum; Lee, Kyung-Hun; Yang, Yaewon; Kim, Tae-Yong; Lim, Jee Min; Park, So Jung; Nam, Hyun-Jin; Kim, Jung Eun; Song, Sang-Hyun; Han, Sae-Won; Oh, Do-Youn; Kim, Jee Hyun; Kim, Tae-You; Hangauer, David; Lau, Johnson Yiu-Nam; Im, Kyongok; Lee, Dong Soon; Bang, Yung-Jue; Im, Seock-Ah

2017-07-01

KX-01 is a novel dual inhibitor of Src and tubulin. Unlike previous Src inhibitors that failed to show clinical benefit during treatment of breast cancer, KX-01 can potentially overcome the therapeutic limitations of current Src inhibitors through inhibition of both Src and tubulin. The present study further evaluates the activity and mechanism of KX-01 in vitro and in vivo . The antitumor effect of KX-01 in triple negative breast cancer (TNBC) cell lines was determined by MTT assay. Wound healing and immunofluorescence assays were performed to evaluate the action mechanisms of KX-01. Changes in the cell cycle and molecular changes induced by KX-01 were also evaluated. A MDA-MB-231 mouse xenograft model was used to demonstrate the in vivo effects. KX-01 effectively inhibited the growth of breast cancer cell lines. The expression of phospho-Src and proliferative-signaling molecules were down-regulated in KX-01-sensitive TNBC cell lines. In addition, migration inhibition was observed by wound healing assay. KX-01-induced G2/M cell cycle arrest and increased the aneuploid cell population in KX-01-sensitive cell lines. Multi-nucleated cells were significantly increased after KX-01 treatment. Furthermore, KX-01 effectively delayed tumor growth in a MDA-MB-231 mouse xenograft model. KX-01 effectively inhibited cell growth and migration of TNBC cells. Moreover, this study demonstrated that KX-01 showed antitumor effects through the inhibition of Src signaling and the induction of mitotic catastrophe. The antitumor effects of KX-01 were also demonstrated in vivo using a mouse xenograft model.

Dihydrotestosterone Potentiates EGF-Induced ERK Activation by Inducing SRC in Fetal Lung Fibroblasts

Science.gov (United States)

Smith, Susan M.; Murray, Sandy; Pham, Lucia D.; Minoo, Parviz; Nielsen, Heber C.

2014-01-01

Lung maturation is regulated by interactions between mesenchymal and epithelial cells, and is delayed by androgens. Fibroblast–Type II cell communications are dependent on extracellular signal-regulated kinases (ERK) 1/2 activation by the ErbB receptor ligands epidermal growth factor (EGF), transforming growth factor (TGF)-α, and neuregulin (Nrg). In other tissues, dihydrotestosterone (DHT) has been shown to activate SRC by a novel nontranscriptional mechanism, which phosphorylates EGF receptors to potentiate EGF-induced ERK1/2 activation. This study sought to determine if DHT potentiates EGFR signaling by a nontranscriptional mechanism. Embryonic day (E)17 fetal lung cells were isolated from dams treated with or without DHT since E12. Cells were exposed to 30 ng/ml DHT for periods of 30 minutes to 3 days before being stimulated with 100 ng/ml EGF, TGF-α, or Nrg for up to 30 minutes. Lysates were immunoblotted for ErbB and SRC pathway signaling intermediates. DHT increased ERK1/2 activation by EGF, TGF-α, and Nrg in fibroblasts and Type II cells. Characterization in fibroblasts showed that potentiation of the EGF pathway was significant after 60 minutes of DHT exposure and persisted in the presence of the translational inhibitor cycloheximide. SRC and EGF receptor phosphorylation was increased by DHT, as was EGF-induced SHC1 phosphorylation and subsequent association with GRB2. Finally, SRC silencing, SRC inhibition with PP2, and overexpression of a dominant-negative SRC each prevented DHT from increasing EGF-induced ERK1/2 phosphorylation. These results suggest that DHT activates SRC to potentiate the signaling pathway leading from the EGF receptor to ERK activation in primary fetal lung fibroblasts. PMID:24484548

Src mediates cigarette smoke-induced resistance to tyrosine kinase inhibitors in NSCLC cells.

Science.gov (United States)

Filosto, Simone; Baston, David S; Chung, Samuel; Becker, Cathleen R; Goldkorn, Tzipora

2013-08-01

The EGF receptor (EGFR) is a proto-oncogene commonly dysregulated in several cancers including non-small cell lung carcinoma (NSCLC) and, thus, is targeted for treatment using tyrosine kinase inhibitors (TKI) such as erlotinib. However, despite the efficacy observed in patients with NSCLC harboring oncogenic variants of the EGFR, general ineffectiveness of TKIs in patients with NSCLC who are current and former smokers necessitates identification of novel mechanisms to overcome this phenomenon. Previously, we showed that NSCLC cells harboring either wild-type (WT) EGFR or oncogenic mutant (MT) L858R EGFR become resistant to the effects of TKIs when exposed to cigarette smoke, evidenced by their autophosphorylation and prolonged downstream signaling. Here, we present Src as a target mediating cigarette smoke-induced resistance to TKIs in both WT EGFR- and L858R MT EGFR-expressing NSCLC cells. First, we show that cigarette smoke exposure of A549 cells leads to time-dependent activation of Src, which then abnormally binds to the WT EGFR causing TKI resistance, contrasting previous observations of constitutive binding between inactive Src and TKI-sensitive L858R MT EGFR. Next, we show that Src inhibition restores TKI sensitivity in cigarette smoke-exposed NSCLC cells, preventing EGFR autophosphorylation in the presence of erlotinib. Furthermore, we show that overexpression of a dominant-negative Src (Y527F/K295R) restores TKI sensitivity to A549 exposed to cigarette smoke. Importantly, the TKI resistance that emerges even in cigarette smoke-exposed L858R EGFR-expressing NSCLC cells could be eliminated with Src inhibition. Together, these findings offer new rationale for using Src inhibitors for treating TKI-resistant NSCLC commonly observed in smokers.

DMPD: Regulation of phagocyte migration and recruitment by Src-family kinases. [Dynamic Macrophage Pathway CSML Database

Lifescience Database Archive (English)

Full Text Available 18385944 Regulation of phagocyte migration and recruitment by Src-family kinases. B...how Regulation of phagocyte migration and recruitment by Src-family kinases. PubmedID 18385944 Title Regulat...ion of phagocyte migration and recruitment by Src-family kinases. Authors Baruzzi

Acetylation within the N- and C-Terminal Domains of Src Regulates Distinct Roles of STAT3-Mediated Tumorigenesis.

Science.gov (United States)

Huang, Chao; Zhang, Zhe; Chen, Lihan; Lee, Hank W; Ayrapetov, Marina K; Zhao, Ting C; Hao, Yimei; Gao, Jinsong; Yang, Chunzhang; Mehta, Gautam U; Zhuang, Zhengping; Zhang, Xiaoren; Hu, Guohong; Chin, Y Eugene

2018-06-01

Posttranslational modifications of mammalian c-Src N-terminal and C-terminal domains regulate distinct functions. Myristoylation of G 2 controls its cell membrane association and phosphorylation of Y419/Y527 controls its activation or inactivation, respectively. We provide evidence that Src-cell membrane association-dissociation and catalytic activation-inactivation are both regulated by acetylation. In EGF-treated cells, CREB binding protein (CBP) acetylates an N-terminal lysine cluster (K5, K7, and K9) of c-Src to promote dissociation from the cell membrane. CBP also acetylates the C-terminal K401, K423, and K427 of c-Src to activate intrinsic kinase activity for STAT3 recruitment and activation. N-terminal domain phosphorylation (Y14, Y45, and Y68) of STAT3 by c-Src activates transcriptionally active dimers of STAT3. Moreover, acetyl-Src translocates into nuclei, where it forms the Src-STAT3 enhanceosome for gene regulation and cancer cell proliferation. Thus, c-Src acetylation in the N-terminal and C-terminal domains play distinct roles in Src activity and regulation. Significance: CBP-mediated acetylation of lysine clusters in both the N-terminal and C-terminal regions of c-Src provides additional levels of control over STAT3 transcriptional activity. Cancer Res; 78(11); 2825-38. ©2018 AACR . ©2018 American Association for Cancer Research.

Erratum to: Fertilization of SRC Willow

DEFF Research Database (Denmark)

Sevel, L; Ingerslev, Morten; Nord-Larsen, Thomas

2014-01-01

impacts of different doses of mineral fertilizer, manure and sewage sludge in a commercially grown SRC willow stand. We examined macro nutrient and heavy metal leaching rates and calculated element balances to evaluate the environmental impact. Growth responses were reported in a former paper (Sevel et al...

76 FR 3653 - Alaska Region's Subsistence Resource Commission (SRC) Program; Public Meeting

Science.gov (United States)

2011-01-20

... subsistence management issues. The NPS SRC program is authorized under Title VIII, Section 808 of the Alaska...: 1. Call to order. 2. SRC Roll Call and Confirmation of Quorum. 3. Welcome and Introductions. 4... Board of Game Update. 12. Old Business. a. Subsistence Uses of Horns, Antlers, Bones and Plants EA...

Regulation of mTORC1 Signaling by Src Kinase Activity Is Akt1-Independent in RSV-Transformed Cells

Directory of Open Access Journals (Sweden)

Martina Vojtěchová

2008-02-01

Full Text Available Increased activity of the Src tyrosine protein kinase that has been observed in a large number of human malignancies appears to be a promising target for drug therapy. In the present study, a critical role of the Src activity in the deregulation of mTOR signaling pathway in Rous sarcoma virus (RSV-transformed hamster fibroblasts, H19 cells, was shown using these cells treated with the Src-specific inhibitor, SU6656, and clones of fibroblasts expressing either the active Src or the dominant-negative Src kinase-dead mutant. Disruption of the Src kinase activity results in substantial reduction of the phosphorylation and activity of the Akt/protein kinase B (PKB, phosphorylation of tuberin (TSC2, mammalian target of rapamycin (mTOR, S6K1, ribosomal protein S6, and eukaryotic initiation factor 4E-binding protein 4E-BP1. The ectopic, active Akt1 that was expressed in Src-deficient cells significantly enhanced phosphorylation of TSC2 in these cells, but it failed to activate the inhibited components of the mTOR pathway that are downstream of TSC2. The data indicate that the Src kinase activity is essential for the activity of mTOR-dependent signaling pathway and suggest that mTOR targets may be controlled by Src independently of Akt1/TSC2 cascade in cells expressing hyperactive Src protein. These observations might have an implication in drug resistance to mTOR inhibitor-based cancer therapy in certain cell types.

SLAP displays tumour suppressor functions in colorectal cancer via destabilization of the SRC substrate EPHA2

Science.gov (United States)

Naudin, Cécile; Sirvent, Audrey; Leroy, Cédric; Larive, Romain; Simon, Valérie; Pannequin, Julie; Bourgaux, Jean-François; Pierre, Josiane; Robert, Bruno; Hollande, Frédéric; Roche, Serge

2014-01-01

The adaptor SLAP is a negative regulator of receptor signalling in immune cells but its role in human cancer is ill defined. Here we report that SLAP is abundantly expressed in healthy epithelial intestine but strongly downregulated in 50% of colorectal cancer. SLAP overexpression suppresses cell tumorigenicity and invasiveness while SLAP silencing enhances these transforming properties. Mechanistically, SLAP controls SRC/EPHA2/AKT signalling via destabilization of the SRC substrate and receptor tyrosine kinase EPHA2. This activity is independent from CBL but requires SLAP SH3 interaction with the ubiquitination factor UBE4A and SLAP SH2 interaction with pTyr594-EPHA2. SRC phosphorylates EPHA2 on Tyr594, thus creating a feedback loop that promotes EPHA2 destruction and thereby self-regulates its transforming potential. SLAP silencing enhances SRC oncogenicity and sensitizes colorectal tumour cells to SRC inhibitors. Collectively, these data establish a tumour-suppressive role for SLAP in colorectal cancer and a mechanism of SRC oncogenic induction through stabilization of its cognate substrates.

Structure-based design of an osteoclast-selective, nonpeptide Src homology 2 inhibitor with in vivo antiresorptive activity

Science.gov (United States)

Shakespeare, William; Yang, Michael; Bohacek, Regine; Cerasoli, Franklin; Stebbins, Karin; Sundaramoorthi, Raji; Azimioara, Mihai; Vu, Chi; Pradeepan, Selvi; Metcalf, Chester; Haraldson, Chad; Merry, Taylor; Dalgarno, David; Narula, Surinder; Hatada, Marcos; Lu, Xiaode; van Schravendijk, Marie Rose; Adams, Susan; Violette, Shelia; Smith, Jeremy; Guan, Wei; Bartlett, Catherine; Herson, Jay; Iuliucci, John; Weigele, Manfred; Sawyer, Tomi

2000-01-01

Targeted disruption of the pp60src (Src) gene has implicated this tyrosine kinase in osteoclast-mediated bone resorption and as a therapeutic target for the treatment of osteoporosis and other bone-related diseases. Herein we describe the discovery of a nonpeptide inhibitor (AP22408) of Src that demonstrates in vivo antiresorptive activity. Based on a cocrystal structure of the noncatalytic Src homology 2 (SH2) domain of Src complexed with citrate [in the phosphotyrosine (pTyr) binding pocket], we designed 3′,4′-diphosphonophenylalanine (Dpp) as a pTyr mimic. In addition to its design to bind Src SH2, the Dpp moiety exhibits bone-targeting properties that confer osteoclast selectivity, hence minimizing possible undesired effects on other cells that have Src-dependent activities. The chemical structure AP22408 also illustrates a bicyclic template to replace the post-pTyr sequence of cognate Src SH2 phosphopeptides such as Ac-pTyr-Glu-Glu-Ile (1). An x-ray structure of AP22408 complexed with Lck (S164C) SH2 confirmed molecular interactions of both the Dpp and bicyclic template of AP22408 as predicted from molecular modeling. Relative to the cognate phosphopeptide, AP22408 exhibits significantly increased Src SH2 binding affinity (IC50 = 0.30 μM for AP22408 and 5.5 μM for 1). Furthermore, AP22408 inhibits rabbit osteoclast-mediated resorption of dentine in a cellular assay, exhibits bone-targeting properties based on a hydroxyapatite adsorption assay, and demonstrates in vivo antiresorptive activity in a parathyroid hormone-induced rat model. PMID:10944210

Distinct protease pathways control cell shape and apoptosis in v-src-transformed quail neuroretina cells

International Nuclear Information System (INIS)

Neel, Benjamin D.; Aouacheria, Abdel; Nouvion, Anne-Laure; Ronot, Xavier; Gillet, Germain

2005-01-01

Intracellular proteases play key roles in cell differentiation, proliferation and apoptosis. In nerve cells, little is known about their relative contribution to the pathways which control cell physiology, including cell death. Neoplastic transformation of avian neuroretina cells by p60 v-src tyrosine kinase results in dramatic morphological changes and deregulation of apoptosis. To identify the proteases involved in the cellular response to p60 v-src , we evaluated the effect of specific inhibitors of caspases, calpains and the proteasome on cell shape changes and apoptosis induced by p60 v-src inactivation in quail neuroretina cells transformed by tsNY68, a thermosensitive strain of Rous sarcoma virus. We found that the ubiquitin-proteasome pathway is recruited early after p60 v-src inactivation and is critical for morphological changes, whereas caspases are essential for cell death. This study provides evidence that distinct intracellular proteases are involved in the control of the morphology and fate of v-src-transformed cells

From Provenance Standards and Tools to Queries and Actionable Provenance

Science.gov (United States)

Ludaescher, B.

2017-12-01

The W3C PROV standard provides a minimal core for sharing retrospective provenance information for scientific workflows and scripts. PROV extensions such as DataONE's ProvONE model are necessary for linking runtime observables in retrospective provenance records with conceptual-level prospective provenance information, i.e., workflow (or dataflow) graphs. Runtime provenance recorders, such as DataONE's RunManager for R, or noWorkflow for Python capture retrospective provenance automatically. YesWorkflow (YW) is a toolkit that allows researchers to declare high-level prospective provenance models of scripts via simple inline comments (YW-annotations), revealing the computational modules and dataflow dependencies in the script. By combining and linking both forms of provenance, important queries and use cases can be supported that neither provenance model can afford on its own. We present existing and emerging provenance tools developed for the DataONE and SKOPE (Synthesizing Knowledge of Past Environments) projects. We show how the different tools can be used individually and in combination to model, capture, share, query, and visualize provenance information. We also present challenges and opportunities for making provenance information more immediately actionable for the researchers who create it in the first place. We argue that such a shift towards "provenance-for-self" is necessary to accelerate the creation, sharing, and use of provenance in support of transparent, reproducible computational and data science.

Trihydrophobin 1 Phosphorylation by c-Src Regulates MAPK/ERK Signaling and Cell Migration

Science.gov (United States)

Wu, Weibin; Sun, Zhichao; Wu, Jingwen; Peng, Xiaomin; Gan, Huacheng; Zhang, Chunyi; Ji, Lingling; Xie, Jianhui; Zhu, Haiyan; Ren, Shifang

2012-01-01

c-Src activates Ras-MAPK/ERK signaling pathway and regulates cell migration, while trihydrophobin 1 (TH1) inhibits MAPK/ERK activation and cell migration through interaction with A-Raf and PAK1 and inhibiting their kinase activities. Here we show that c-Src interacts with TH1 by GST-pull down assay, coimmunoprecipitation and confocal microscopy assay. The interaction leads to phosphorylation of TH1 at Tyr-6 in vivo and in vitro. Phosphorylation of TH1 decreases its association with A-Raf and PAK1. Further study reveals that Tyr-6 phosphorylation of TH1 reduces its inhibition on MAPK/ERK signaling, enhances c-Src mediated cell migration. Moreover, induced tyrosine phosphorylation of TH1 has been found by EGF and estrogen treatments. Taken together, our findings demonstrate a novel mechanism for the comprehensive regulation of Ras/Raf/MEK/ERK signaling and cell migration involving tyrosine phosphorylation of TH1 by c-Src. PMID:22238675

SP600125 Induces Src and Type I IGF Receptor Phosphorylation Independent of JNK

Directory of Open Access Journals (Sweden)

Qingbin Kong

2014-09-01

Full Text Available c-Jun N-terminal kinases (JNK are members of the mitogen-activated protein kinase (MAPK family that have important roles in signal transduction. The small molecule SP600125 is widely used in biochemical studies as a JNK inhibitor. However, recent studies indicate that SP600125 may also act independent of JNK. Here, we report that SP600125 can induce Src, type I insulin-like growth factor receptor (IGF-IR, Akt and Erk1/2 phosphorylation. Notably, these effects are independent of its inhibition of JNK. Inhibition of Src abrogates the stimulation of IGF-IR, Akt and Erk1/2 phosphorylation. IGF-IR knockdown blunts the induction of both Akt and Erk1/2 phosphorylation by SP600125. Moreover, combination of SP600125 and the Src inhibitor saracatinib synergistically inhibits cell proliferation. We conclude that SP600125 can activate Src-IGF-IR-Akt/Erk1/2 signaling pathways independent of JNK.

Adaptor protein GRB2 promotes Src tyrosine kinase activation and podosomal organization by protein-tyrosine phosphatase ϵ in osteoclasts.

Science.gov (United States)

Levy-Apter, Einat; Finkelshtein, Eynat; Vemulapalli, Vidyasiri; Li, Shawn S-C; Bedford, Mark T; Elson, Ari

2014-12-26

The non-receptor isoform of protein-tyrosine phosphatase ϵ (cyt-PTPe) supports adhesion of bone-resorbing osteoclasts by activating Src downstream of integrins. Loss of cyt-PTPe reduces Src activity in osteoclasts, reduces resorption of mineralized matrix both in vivo and in cell culture, and induces mild osteopetrosis in young female PTPe KO mice. Activation of Src by cyt-PTPe is dependent upon this phosphatase undergoing phosphorylation at its C-terminal Tyr-638 by partially active Src. To understand how cyt-PTPe activates Src, we screened 73 Src homology 2 (SH2) domains for binding to Tyr(P)-638 of cyt-PTPe. The SH2 domain of GRB2 bound Tyr(P)-638 of cyt-PTPe most prominently, whereas the Src SH2 domain did not bind at all, suggesting that GRB2 may link PTPe with downstream molecules. Further studies indicated that GRB2 is required for activation of Src by cyt-PTPe in osteoclast-like cells (OCLs) in culture. Overexpression of GRB2 in OCLs increased activating phosphorylation of Src at Tyr-416 and of cyt-PTPe at Tyr-638; opposite results were obtained when GRB2 expression was reduced by shRNA or by gene inactivation. Phosphorylation of cyt-PTPe at Tyr-683 and its association with GRB2 are integrin-driven processes in OCLs, and cyt-PTPe undergoes autodephosphorylation at Tyr-683, thus limiting Src activation by integrins. Reduced GRB2 expression also reduced the ability of bone marrow precursors to differentiate into OCLs and reduced the fraction of OCLs in which podosomal adhesion structures assume organization typical of active, resorbing cells. We conclude that GRB2 physically links cyt-PTPe with Src and enables cyt-PTPe to activate Src downstream of activated integrins in OCLs. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

Src kinases in chondrosarcoma chemoresistance and migration: dasatinib sensitises to doxorubicin in TP53 mutant cells

Science.gov (United States)

van Oosterwijk, J G; van Ruler, M A J H; Briaire-de Bruijn, I H; Herpers, B; Gelderblom, H; van de Water, B; Bovée, J V M G

2013-01-01

Background: Chondrosarcomas are malignant cartilage-forming tumours of bone. Because of their resistance to conventional chemotherapy and radiotherapy, currently no treatment strategies exist for unresectable and metastatic chondrosarcoma. Previously, PI3K/AKT/GSK3β and Src kinase pathways were shown to be activated in chondrosarcoma cell lines. Our aim was to investigate the role of these kinases in chemoresistance and migration in chondrosarcoma in relation to TP53 mutation status. Methods: We used five conventional and three dedifferentiated chondrosarcoma cell lines and investigated the effect of PI3K/AKT/GSK3β pathway inhibition (enzastaurin) and Src pathway inhibition (dasatinib) in chemoresistance using WST assay and live cell imaging with AnnexinV staining. Immunohistochemistry on tissue microarrays (TMAs) containing 157 cartilaginous tumours was performed for Src family members. Migration assays were performed with the RTCA xCelligence System. Results: Src inhibition was found to overcome chemoresistance, to induce apoptosis and to inhibit migration. Cell lines with TP53 mutations responded better to combination therapy than wild-type cell lines (P=0.002). Tissue microarray immunohistochemistry confirmed active Src (pSrc) signalling, with Fyn being most abundantly expressed (76.1%). Conclusion: These results strongly indicate Src family kinases, in particular Fyn, as a potential target for the treatment of inoperable and metastatic chondrosarcomas, and to sensitise for doxorubicin especially in the presence of TP53 mutations. PMID:23922104

FUS-CHOP Promotes Invasion in Myxoid Liposarcoma through a SRC/FAK/RHO/ROCK-Dependent Pathway

Directory of Open Access Journals (Sweden)

Juan Tornin

2018-01-01

Full Text Available Deregulated SRC/FAK signaling leads to enhanced migration and invasion in many types of tumors. In myxoid and round cell liposarcoma (MRCLS, an adipocytic tumor characterized by the expression of the fusion oncogene FUS-CHOP, SRC have been found as one of the most activated kinases. Here we used a cell-of-origin model of MRCLS and an MRCLS cell line to thoroughly characterize the mechanisms of cell invasion induced by FUS-CHOP using in vitro (3D spheroid invasion assays and in vivo (chicken chorioallantoic membrane model approaches. FUS-CHOP expression activated SRC-FAK signaling and increased the invasive ability of MRCLS cells. In addition, FAK expression was found to significantly correlate with tumor aggressiveness in sarcoma patient samples. The involvement of SRC/FAK activation in FUS-CHOP–mediated invasion was further confirmed using the SRC inhibitor dasatinib, the specific FAK inhibitor PF-573228, and FAK siRNA. Notably, dasatinib and PF573228 could also efficiently block the invasion of cancer stem cell subpopulations. Downstream of SRC/FAK signaling, we found that FUS-CHOP expression increases the levels of the RHO/ROCK downstream effector phospho-MLC2 (T18/S19 and that this activation was prevented by dasatinib or PF573228. Moreover, the ROCK inhibitor RKI-1447 was able to completely abolish invasion in FUS-CHOP–expressing cells. These data uncover the involvement of SRC/FAK/RHO/ROCK signaling axis in FUS-CHOP–mediated invasion, thus providing a rationale for testing inhibitors of this pathway as potential novel antimetastatic agents for MRCLS treatment.

The Role of C-SRC Activation in Prostate Tumor Progression

Science.gov (United States)

2006-07-01

cancer cell line PANC -1 and prostrate cancer cell line PC-3 (B2-fold increase relative to control in both cell lines), while the Src inhibitory PP2 blocks...at normoxia in PANC -1 and PC-3 cells, its levels significantly increase in response to hypoxia (B4.5–8-fold induction). Inhibition of endo- genous c...Src activation in PANC -1 and PC-3 cells by PP2 drastically reduced HIF-1a levels to below those levels observed at normoxia (Figure 1a). STAT3 has

Digitalis-induced cell signaling by the sodium pump: on the relation of Src to Na(+)/K(+)-ATPase.

Science.gov (United States)

Gable, Marjorie E; Abdallah, Simon L; Najjar, Sonia M; Liu, Lijun; Askari, Amir

2014-04-18

In addition to performing its essential transport function, the sodium pump also activates multiple cell signaling pathways in response to digitalis drugs such as ouabain. Based mainly on cell-free studies with mixtures of purified Src kinase and Na(+)/K(+)-ATPase, a well-advocated hypothesis on how ouabain initiates the activation of signaling pathways is that there is a preexisting physiological complex of inactive Src bound to the α-subunit of Na(+)/K(+)-ATPase, and that ouabain binding to this subunit disrupts the bound Src and activates it. Because of the published disagreements of the results of such cell-free experiments of two other laboratories, our aim was to attempt the resolution of these discrepancies. We reexamined the effects of ouabain, vanadate, and oligomycin on mixtures of Src, Na(+)/K(+)-ATPase, Mg(2+), and ATP as specified in prior studies; and assayed for Src-418 autophosphorylation as the measure of Src activation. In contrast to the findings of the proponents of the above hypothesis, our results showed similar effects of the three inhibitors of Na(+)/K(+)-ATPase; indicating that Src activation in such experiments is primarily due to the ATP-sparing effect of the ATPase inhibitor on the mixture of two enzymes competing for ATP. We conclude that there is no solid evidence for direct molecular interaction of Src with Na(+)/K(+)-ATPase under physiological conditions. Copyright © 2014 The Authors. Published by Elsevier Inc. All rights reserved.

Src kinase conformational activation: thermodynamics, pathways, and mechanisms.

Directory of Open Access Journals (Sweden)

Sichun Yang

2008-03-01

Full Text Available Tyrosine kinases of the Src-family are large allosteric enzymes that play a key role in cellular signaling. Conversion of the kinase from an inactive to an active state is accompanied by substantial structural changes. Here, we construct a coarse-grained model of the catalytic domain incorporating experimental structures for the two stable states, and simulate the dynamics of conformational transitions in kinase activation. We explore the transition energy landscapes by constructing a structural network among clusters of conformations from the simulations. From the structural network, two major ensembles of pathways for the activation are identified. In the first transition pathway, we find a coordinated switching mechanism of interactions among the alphaC helix, the activation-loop, and the beta strands in the N-lobe of the catalytic domain. In a second pathway, the conformational change is coupled to a partial unfolding of the N-lobe region of the catalytic domain. We also characterize the switching mechanism for the alphaC helix and the activation-loop in detail. Finally, we test the performance of a Markov model and its ability to account for the structural kinetics in the context of Src conformational changes. Taken together, these results provide a broad framework for understanding the main features of the conformational transition taking place upon Src activation.

Specific oncogenic activity of the Src-family tyrosine kinase c-Yes in colon carcinoma cells.

Directory of Open Access Journals (Sweden)

Florence Sancier

Full Text Available c-Yes, a member of the Src tyrosine kinase family, is found highly activated in colon carcinoma but its importance relative to c-Src has remained unclear. Here we show that, in HT29 colon carcinoma cells, silencing of c-Yes, but not of c-Src, selectively leads to an increase of cell clustering associated with a localisation of β-catenin at cell membranes and a reduction of expression of β-catenin target genes. c-Yes silencing induced an increase in apoptosis, inhibition of growth in soft-agar and in mouse xenografts, inhibition of cell migration and loss of the capacity to generate liver metastases in mice. Re-introduction of c-Yes, but not c -Src, restores transforming properties of c-Yes depleted cells. Moreover, we found that c-Yes kinase activity is required for its role in β-catenin localisation and growth in soft agar, whereas kinase activity is dispensable for its role in cell migration. We conclude that c-Yes regulates specific oncogenic signalling pathways important for colon cancer progression that is not shared with c-Src.

Src-family kinases negatively regulate NFAT signaling in resting human T cells.

Directory of Open Access Journals (Sweden)

Alan Baer

Full Text Available T cell signaling is required for activation of both natural and therapeutic T cells including chimeric antigen receptor (CAR T cells. Identification of novel factors and pathways regulating T cell signaling may aid in development of effective T cell therapies. In resting human T cells, the majority of Src-family of tyrosine kinases (SFKs are inactive due to phosphorylation of a conserved carboxy-terminal tyrosine residue. Recently, a pool of enzymatically active SFKs has been identified in resting T cells; however, the significance of these is incompletely understood. Here, we characterized the role of active SFKs in resting human T cells. Pharmacologic inhibition of active SFKs enhanced distal TCR signaling as measured by IL-2 release and CD25 surface expression following TCR-independent activation. Mechanistically, inhibition of the active pool of SFKs induced nuclear translocation of NFAT1, and enhanced NFAT1-dependent signaling in resting T cells. The negative regulation of NFAT1 signaling was in part mediated by the Src-kinase Lck as human T cells lacking Lck had increased levels of nuclear NFAT1 and demonstrated enhanced NFAT1-dependent gene expression. Inhibition of active SFKs in resting primary human T cells also increased nuclear NFAT1 and enhanced NFAT1-dependent signaling. Finally, the calcineurin inhibitor FK506 and Cyclosporin A reversed the effect of SFKs inhibition on NFAT1. Together, these data identified a novel role of SFKs in preventing aberrant NFAT1 activation in resting T cells, and suggest that maintaining this pool of active SFKs in therapeutic T cells may increase the efficacy of T cell therapies.

Src Kinase becomes preferentially associated with the VEGFR, KDR/Flk-1, following VEGF stimulation of vascular endothelial cells

Directory of Open Access Journals (Sweden)

Wang Jing

2002-12-01

Full Text Available Abstract Background The cytoplasmic tyrosine kinase, Src, has been found to play a crucial role in VEGF (vascular endothelial growth factor – dependent vascular permeability involved in angiogenesis. The two main VEGFRs present on vascular endothelial cells are KDR/Flk-1 (kinase insert domain-containing receptor/fetal liver kinase-1 and Flt-1 (Fms-like tyrosine kinase-1. However, to date, it has not been determined which VEGF receptor (VEGFR is involved in binding to and activating Src kinase following VEGF stimulation of the receptors. Results In this report, we demonstrate that Src preferentially associates with KDR/Flk-1 rather than Flt-1 in human umbilical vein endothelial cells (HUVECs, and that VEGF stimulation resulted in an increase of Src activity associated with activated KDR/Flk-1. These findings were determined through immunoprecipitation-kinase experiments and coimmunoprecipitation studies, and were further confirmed by GST-pull-down assays and Far Western studies. However, Fyn and Yes, unlike Src, were found to associate preferentially with Flt-1. Conclusions Thus, Src preferentially associates with KDR/Flk-1, rather than with Flt-1, upon VEGF stimulation in endothelial cells. Our findings further highlight the potential significance of upregulated KDR/Flk-1-associated Src activity in the process of angiogenesis, and help to elucidate more clearly the specific roles and mechanisms involving Src family tyrosine kinase in VEGF-stimulated signal transduction events.

Aggregation by Provenance Types: A Technique for Summarising Provenance Graphs

Directory of Open Access Journals (Sweden)

Luc Moreau

2015-04-01

Full Text Available As users become confronted with a deluge of provenance data, dedicated techniques are required to make sense of this kind of information. We present Aggregation by Provenance Types, a provenance graph analysis that is capable of generating provenance graph summaries. It proceeds by converting provenance paths up to some length k to attributes, referred to as provenance types, and by grouping nodes that have the same provenance types. The summary also includes numeric values representing the frequency of nodes and edges in the original graph. A quantitative evaluation and a complexity analysis show that this technique is tractable; with small values of k, it can produce useful summaries and can help detect outliers. We illustrate how the generated summaries can further be used for conformance checking and visualization.

Dynamically Coupled Residues within the SH2 Domain of FYN Are Key to Unlocking Its Activity.

Science.gov (United States)

Huculeci, Radu; Cilia, Elisa; Lyczek, Agatha; Buts, Lieven; Houben, Klaartje; Seeliger, Markus A; van Nuland, Nico; Lenaerts, Tom

2016-11-01

Src kinase activity is controlled by various mechanisms involving a coordinated movement of kinase and regulatory domains. Notwithstanding the extensive knowledge related to the backbone dynamics, little is known about the more subtle side-chain dynamics within the regulatory domains and their role in the activation process. Here, we show through experimental methyl dynamic results and predicted changes in side-chain conformational couplings that the SH2 structure of Fyn contains a dynamic network capable of propagating binding information. We reveal that binding the phosphorylated tail of Fyn perturbs a residue cluster near the linker connecting the SH2 and SH3 domains of Fyn, which is known to be relevant in the regulation of the activity of Fyn. Biochemical perturbation experiments validate that those residues are essential for inhibition of Fyn, leading to a gain of function upon mutation. These findings reveal how side-chain dynamics may facilitate the allosteric regulation of the different members of the Src kinase family. Copyright © 2016 Elsevier Ltd. All rights reserved.

The Haemophilus ducreyi LspA1 protein inhibits phagocytosis by using a new mechanism involving activation of C-terminal Src kinase.

Science.gov (United States)

Dodd, Dana A; Worth, Randall G; Rosen, Michael K; Grinstein, Sergio; van Oers, Nicolai S C; Hansen, Eric J

2014-05-20

Haemophilus ducreyi causes chancroid, a sexually transmitted infection. A primary means by which this pathogen causes disease involves eluding phagocytosis; however, the molecular basis for this escape mechanism has been poorly understood. Here, we report that the LspA virulence factors of H. ducreyi inhibit phagocytosis by stimulating the catalytic activity of C-terminal Src kinase (Csk), which itself inhibits Src family protein tyrosine kinases (SFKs) that promote phagocytosis. Inhibitory activity could be localized to a 37-kDa domain (designated YL2) of the 456-kDa LspA1 protein. The YL2 domain impaired ingestion of IgG-opsonized targets and decreased levels of active SFKs when expressed in mammalian cells. YL2 contains tyrosine residues in two EPIYG motifs that are phosphorylated in mammalian cells. These tyrosine residues were essential for YL2-based inhibition of phagocytosis. Csk was identified as the predominant mammalian protein interacting with YL2, and a dominant-negative Csk rescued phagocytosis in the presence of YL2. Purified Csk phosphorylated the tyrosines in the YL2 EPIYG motifs. Phosphorylated YL2 increased Csk catalytic activity, resulting in positive feedback, such that YL2 can be phosphorylated by the same kinase that it activates. Finally, we found that the Helicobacter pylori CagA protein also inhibited phagocytosis in a Csk-dependent manner, raising the possibility that this may be a general mechanism among diverse bacteria. Harnessing Csk to subvert the Fcγ receptor (FcγR)-mediated phagocytic pathway represents a new bacterial mechanism for circumventing a crucial component of the innate immune response and may potentially affect other SFK-involved cellular pathways. Phagocytosis is a critical component of the immune system that enables pathogens to be contained and cleared. A number of bacterial pathogens have developed specific strategies to either physically evade phagocytosis or block the intracellular signaling required for

Focal adhesion kinase-dependent focal adhesion recruitment of SH2 domains directs SRC into focal adhesions to regulate cell adhesion and migration.

Science.gov (United States)

Wu, Jui-Chung; Chen, Yu-Chen; Kuo, Chih-Ting; Wenshin Yu, Helen; Chen, Yin-Quan; Chiou, Arthur; Kuo, Jean-Cheng

2015-12-18

Directed cell migration requires dynamical control of the protein complex within focal adhesions (FAs) and this control is regulated by signaling events involving tyrosine phosphorylation. We screened the SH2 domains present in tyrosine-specific kinases and phosphatases found within FAs, including SRC, SHP1 and SHP2, and examined whether these enzymes transiently target FAs via their SH2 domains. We found that the SRC_SH2 domain and the SHP2_N-SH2 domain are associated with FAs, but only the SRC_SH2 domain is able to be regulated by focal adhesion kinase (FAK). The FAK-dependent association of the SRC_SH2 domain is necessary and sufficient for SRC FA targeting. When the targeting of SRC into FAs is inhibited, there is significant suppression of SRC-mediated phosphorylation of paxillin and FAK; this results in an inhibition of FA formation and maturation and a reduction in cell migration. This study reveals an association between FAs and the SRC_SH2 domain as well as between FAs and the SHP2_N-SH2 domains. This supports the hypothesis that the FAK-regulated SRC_SH2 domain plays an important role in directing SRC into FAs and that this SRC-mediated FA signaling drives cell migration.

Structural Analysis of DFG-in and DFG-out Dual Src-Abl Inhibitors Sharing a Common Vinyl Purine Template

Energy Technology Data Exchange (ETDEWEB)

Zhou, Tianjun; Commodore, Lois; Huang, Wei-Sheng; Wang, Yihan; Sawyer, Tomi K.; Shakespeare, William C.; Clackson, Tim; Zhu, Xiaotian; Dalgarno, David C. (ARIAD)

2010-09-30

Bcr-Abl is the oncogenic protein tyrosine kinase responsible for chronic myeloid leukemia (CML). Treatment of the disease with imatinib (Gleevec) often results in drug resistance via kinase mutations at the advanced phases of the disease, which has necessitated the development of new mutation-resistant inhibitors, notably against the T315I gatekeeper mutation. As part of our efforts to discover such mutation resistant Abl inhibitors, we have focused on optimizing purine template kinase inhibitors, leading to the discovery of potent DFG-in and DFG-out series of Abl inhibitors that are also potent Src inhibitors. Here we present crystal structures of Abl bound by two such inhibitors, based on a common N9-arenyl purine, and that represent both DFG-in and -out binding modes. In each structure the purine template is bound deeply in the adenine pocket and the novel vinyl linker forms a non-classical hydrogen bond to the gatekeeper residue, Thr315. Specific template substitutions promote either a DFG-in or -out binding mode, with the kinase binding site adjusting to optimize molecular recognition. Bcr-Abl T315I mutant kinase is resistant to all currently marketed Abl inhibitors, and is the focus of intense drug discovery efforts. Notably, our DFG-out inhibitor, AP24163, exhibits modest activity against this mutant, illustrating that this kinase mutant can be inhibited by DFG-out class inhibitors. Furthermore our DFG-out inhibitor exhibits dual Src-Abl activity, absent from the prototypical DFG-out inhibitor, imatinib as well as its analog, nilotinib. The data presented here provides structural guidance for the further design of novel potent DFG-out class inhibitors against Src, Abl and Abl T315I mutant kinases.

c-Src activation through a TrkA and c-Src interaction is essential for cell proliferation and hematological malignancies

International Nuclear Information System (INIS)

Kim, Min Soo; Kim, Gyoung Mi; Choi, Yun-Jeong; Kim, Hye Joung; Kim, Yoo-Jin; Jin, Wook

2013-01-01

Highlights: •TrkA was mainly present in other types of leukemia including AML. •TrkA enhances the survival of leukemia by activation of PI3K/Akt pathway. •TrkA induced significant hematological malignancies by inducing PLK-1 and Twist-1. •TrkA acted as a key regulator of leukemogenesis and survival through c-Src activation. -- Abstract: Although the kinase receptor TrkA may play an important role in acute myeloid leukemia (AML), its involvement in other types of leukemia has not been reported. Furthermore, how it contributes to leukemogenesis is unknown. Here, we describe a molecular network that is important for TrkA function in leukemogenesis. We found that TrkA is frequently overexpressed in other types of leukemia such as acute lymphoblastic leukemia (ALL), chronic myelogenous leukemia (CML), and myelodysplastic syndrome (MDS) including AML. In addition, TrkA was overexpressed in patients with MDS or secondary AML evolving from MDS. TrkA induced significant hematological malignancies by inducing PLK-1 and Twist-1, and enhanced survival and proliferation of leukemia, which was correlated with activation of the phosphatidylinositol 3-kinase/Akt/mTOR pathway. Moreover, endogenous TrkA associated with c-Src complexes was detected in leukemia. Suppression of c-Src activation by TrkA resulted in markedly decreased expression of PLK-1 and Twist-1 via suppressed activation of Akt/mTOR cascades. These data suggest that TrkA plays a key role in leukemogenesis and reveal an unexpected physiological role for TrkA in the pathogenesis of leukemia. These data have important implications for understanding various hematological malignancies

c-Src activation through a TrkA and c-Src interaction is essential for cell proliferation and hematological malignancies

Energy Technology Data Exchange (ETDEWEB)

Kim, Min Soo; Kim, Gyoung Mi; Choi, Yun-Jeong [Department of Molecular Medicine, School of Medicine, Gachon University, Incheon 406-840 (Korea, Republic of); Kim, Hye Joung [Department of Hematology, Catholic Blood and Marrow Transplantation Center, Seoul St. Mary’s Hospital, College of Medicine, The Catholic University of Korea, Seoul 137-701 (Korea, Republic of); Kim, Yoo-Jin, E-mail: yoojink@catholic.ac.kr [Department of Hematology, Catholic Blood and Marrow Transplantation Center, Seoul St. Mary’s Hospital, College of Medicine, The Catholic University of Korea, Seoul 137-701 (Korea, Republic of); Jin, Wook, E-mail: jinwo@gachon.ac.kr [Department of Molecular Medicine, School of Medicine, Gachon University, Incheon 406-840 (Korea, Republic of); Gachon Medical Research Institute, Gil Medical Center, Incheon 405-760 (Korea, Republic of)

2013-11-15

Highlights: •TrkA was mainly present in other types of leukemia including AML. •TrkA enhances the survival of leukemia by activation of PI3K/Akt pathway. •TrkA induced significant hematological malignancies by inducing PLK-1 and Twist-1. •TrkA acted as a key regulator of leukemogenesis and survival through c-Src activation. -- Abstract: Although the kinase receptor TrkA may play an important role in acute myeloid leukemia (AML), its involvement in other types of leukemia has not been reported. Furthermore, how it contributes to leukemogenesis is unknown. Here, we describe a molecular network that is important for TrkA function in leukemogenesis. We found that TrkA is frequently overexpressed in other types of leukemia such as acute lymphoblastic leukemia (ALL), chronic myelogenous leukemia (CML), and myelodysplastic syndrome (MDS) including AML. In addition, TrkA was overexpressed in patients with MDS or secondary AML evolving from MDS. TrkA induced significant hematological malignancies by inducing PLK-1 and Twist-1, and enhanced survival and proliferation of leukemia, which was correlated with activation of the phosphatidylinositol 3-kinase/Akt/mTOR pathway. Moreover, endogenous TrkA associated with c-Src complexes was detected in leukemia. Suppression of c-Src activation by TrkA resulted in markedly decreased expression of PLK-1 and Twist-1 via suppressed activation of Akt/mTOR cascades. These data suggest that TrkA plays a key role in leukemogenesis and reveal an unexpected physiological role for TrkA in the pathogenesis of leukemia. These data have important implications for understanding various hematological malignancies.

Differential Requirements for Src-Family Kinases in SYK or ZAP70-Mediated SLP-76 Phosphorylation in Lymphocytes

Directory of Open Access Journals (Sweden)

Frank Fasbender

2017-07-01

Full Text Available In a synthetic biology approach using Schneider (S2 cells, we show that SLP-76 is directly phosphorylated at tyrosines Y113 and Y128 by SYK in the presence of ITAM-containing adapters such as CD3ζ, DAP12, or FcεRγ. This phosphorylation was dependent on at least one functional ITAM and a functional SH2 domain within SYK. Inhibition of Src-kinases by inhibitors PP1 and PP2 did not reduce SLP-76 phosphorylation in S2 cells, suggesting an ITAM and SYK dependent, but Src-kinase independent signaling pathway. This direct ITAM/SYK/SLP-76 signaling pathway therefore differs from previously described ITAM signaling. However, the SYK-family kinase ZAP70 required the additional co-expression of the Src-family kinases Fyn or Lck to efficiently phosphorylate SLP-76 in S2 cells. This difference in Src-family kinase dependency of SYK versus ZAP70-mediated ITAM-based signaling was further demonstrated in human lymphocytes. ITAM signaling in ZAP70-expressing T cells was dependent on the activity of Src-family kinases. In contrast, Src-family kinases were partially dispensable for ITAM signaling in SYK-expressing B cells or in natural killer cells, which express SYK and ZAP70. This demonstrates that SYK can signal using a Src-kinase independent ITAM-based signaling pathway, which may be involved in calibrating the threshold for lymphocyte activation.

Coupled motions in the SH2 and kinase domains of Csk control Src phosphorylation.

Science.gov (United States)

Wong, Lilly; Lieser, Scot A; Miyashita, Osamu; Miller, Meghan; Tasken, Kjetil; Onuchic, Josè N; Adams, Joseph A; Woods, Virgil L; Jennings, Patricia A

2005-08-05

The C-terminal Src kinase (Csk) phosphorylates and down-regulates Src family tyrosine kinases. The Csk-binding protein (Cbp) localizes Csk close to its substrates at the plasma membrane, and increases the specific activity of the kinase. To investigate this long-range catalytic effect, the phosphorylation of Src and the conformation of Csk were investigated in the presence of a high-affinity phosphopeptide derived from Cbp. This peptide binds tightly to the SH2 domain and enhances Src recognition (lowers K(m)) by increasing the apparent phosphoryl transfer rate in the Csk active site, a phenomenon detected in rapid quench flow experiments. Previous studies demonstrated that the regulation of Csk activity is linked to conformational changes in the enzyme that can be probed with hydrogen-deuterium exchange methods. We show that the Cbp peptide impacts deuterium incorporation into its binding partner (the SH2 domain), and into the SH2-kinase linker and several sequences in the kinase domain, including the glycine-rich loop in the active site. These findings, along with computational data from normal mode analyses, suggest that the SH2 domain moves in a cantilever fashion with respect to the small lobe of the kinase domain, ordering the active site for catalysis. The binding of a small Cbp-derived peptide to the SH2 domain of Csk modifies these motions, enhancing Src recognition.

Atypical protein kinase C activity is required for extracellular matrix degradation and invasion by Src-transformed cells.

Science.gov (United States)

Rodriguez, Elena M; Dunham, Elizabeth E; Martin, G Steven

2009-10-01

Atypical protein kinase C (aPKC) isoforms have been shown to mediate Src-dependent signaling in response to growth factor stimulation. To determine if aPKC activity contributes to the transformed phenotype of cells expressing oncogenic Src, we have examined the activity and function of aPKCs in 3T3 cells expressing viral Src (v-Src). aPKC activity and tyrosine phosphorylation were found to be elevated in some but not all clones of mouse fibroblasts expressing v-Src. aPKC activity was inhibited either by addition of a membrane-permeable pseudosubstrate, by expression of a dominant-negative aPKC, or by RNAi-mediated knockdown of specific aPKC isoforms. aPKC activity contributes to morphological transformation and stress fiber disruption, and is required for migration of Src-transformed cells and for their ability to polarize at the edge of a monolayer. The lambda isoform of aPKC is specifically required for invasion through extracellular matrix in Boyden chamber assays and for degradation of the extracellular matrix in in situ zymography assays. Tyrosine phosphorylation of aPKClambda is required for its ability to promote cell invasion. The defect in invasion upon aPKC inhibition appears to result from a defect in the assembly and/or function of podosomes, invasive adhesions on the ventral surface of the cell that are sites of protease secretion. aPKC was also found to localize to podosomes of v-Src transformed cells, suggesting a direct role for aPKC in podosome assembly and/or function. We conclude that basal or elevated aPKC activity is required for the ability of Src-transformed cells to degrade and invade the extracellular matrix. Copyright 2009 Wiley-Liss, Inc.

Selective Targeting of SH2 Domain–Phosphotyrosine Interactions of Src Family Tyrosine Kinases with Monobodies

Energy Technology Data Exchange (ETDEWEB)

Kükenshöner, Tim; Schmit, Nadine Eliane; Bouda, Emilie; Sha, Fern; Pojer, Florence; Koide, Akiko; Seeliger, Markus; Koide, Shohei; Hantschel, Oliver

2017-05-01

The binding of Src-homology 2 (SH2) domains to phosphotyrosine (pY) sites is critical for the autoinhibition and substrate recognition of the eight Src family kinases (SFKs). The high sequence conservation of the 120 human SH2 domains poses a significant challenge to selectively perturb the interactions of even the SFK SH2 family against the rest of the SH2 domains. We have developed synthetic binding proteins, termed monobodies, for six of the SFK SH2 domains with nanomolar affinity. Most of these monobodies competed with pY ligand binding and showed strong selectivity for either the SrcA (Yes, Src, Fyn, Fgr) or SrcB subgroup (Lck, Lyn, Blk, Hck). Interactome analysis of intracellularly expressed monobodies revealed that they bind SFKs but no other SH2-containing proteins. Three crystal structures of monobody–SH2 complexes unveiled different and only partly overlapping binding modes, which rationalized the observed selectivity and enabled structure-based mutagenesis to modulate inhibition mode and selectivity. In line with the critical roles of SFK SH2 domains in kinase autoinhibition and T-cell receptor signaling, monobodies binding the Src and Hck SH2 domains selectively activated respective recombinant kinases, whereas an Lck SH2-binding monobody inhibited proximal signaling events downstream of the T-cell receptor complex. Our results show that SFK SH2 domains can be targeted with unprecedented potency and selectivity using monobodies. They are excellent tools for dissecting SFK functions in normal development and signaling and to interfere with aberrant SFK signaling networks in cancer cells.

Estrogen Receptor Folding Modulates cSrc Kinase SH2 Interaction via a Helical Binding Mode

NARCIS (Netherlands)

Nieto, Lidia; Tharun, Inga M; Balk, Mark; Wienk, Hans; Boelens, Rolf; Ottmann, Christian; Milroy, Lech-Gustav; Brunsveld, Luc

2015-01-01

The estrogen receptors (ERs) feature, next to their transcriptional role, important nongenomic signaling actions, with emerging clinical relevance. The Src Homology 2 (SH2) domain mediated interaction between cSrc kinase and ER plays a key role in this; however the molecular determinants of this

Estrogen receptor folding modulates cSrc kinase SH2 interaction via a helical binding mode

NARCIS (Netherlands)

Nieto, L.; Tharun, I.M.; Balk, M.; Wienk, H.; Boelens, R.; Ottmann, C.; Milroy, L.-G.; Brunsveld, L.

2015-01-01

The estrogen receptors (ERs) feature, next to their transcriptional role, important nongenomic signaling actions, with emerging clinical relevance. The Src Homology 2 (SH2) domain mediated interaction between cSrc kinase and ER plays a key role in this; however the molecular determinants of this

The Role of Src in Mammary Epithelial Tumorigenesis

National Research Council Canada - National Science Library

Kusdra, Leonard

2007-01-01

...') similar to the physiological lobular-aveoli structures found in the mammary tissue. Additionally, more invasive carcinoma cells (MDA-MB-231 cells) whereby Src signaling was pharmacologically or genetically inhibited were unable to form actin-rich invasive structures in 3D-rBM culture.

Roles of the SH2 and SH3 domains in the regulation of neuronal Src kinase functions.

Science.gov (United States)

Groveman, Bradley R; Xue, Sheng; Marin, Vedrana; Xu, Jindong; Ali, Mohammad K; Bienkiewicz, Ewa A; Yu, Xian-Min

2011-02-01

Previous studies demonstrated that intra-domain interactions between Src family kinases (SFKs), stabilized by binding of the phosphorylated C-terminus to the SH2 domain and/or binding of the SH2 kinase linker to the SH3 domain, lock the molecules in a closed conformation, disrupt the kinase active site, and inactivate SFKs. Here we report that the up-regulation of N-methyl-D-aspartate receptors (NMDARs) induced by expression of constitutively active neuronal Src (n-Src), in which the C-terminus tyrosine is mutated to phenylalanine (n-Src/Y535F), is significantly reduced by dysfunctions of the SH2 and/or SH3 domains of the protein. Furthermore, we found that dysfunctions of SH2 and/or SH3 domains reduce auto-phosphorylation of the kinase activation loop, depress kinase activity, and decrease NMDAR phosphorylation. The SH2 domain plays a greater regulatory role than the SH3 domain. Our data also show that n-Src binds directly to the C-terminus of the NMDAR NR2A subunit in vitro, with a K(D) of 108.2 ± 13.3 nM. This binding is not Src kinase activity-dependent, and dysfunctions of the SH2 and/or SH3 domains do not significantly affect the binding. These data indicate that the SH2 and SH3 domains may function to promote the catalytic activity of active n-Src, which is important in the regulation of NMDAR functions. © 2010 The Authors Journal compilation © 2010 FEBS.

Report of the SRC working party on databases and database management systems

International Nuclear Information System (INIS)

Crennell, K.M.

1980-10-01

An SRC working party, set up to consider the subject of support for databases within the SRC, were asked to identify interested individuals and user communities, establish which features of database management systems they felt were desirable, arrange demonstrations of possible systems and then make recommendations for systems, funding and likely manpower requirements. This report describes the activities and lists the recommendations of the working party and contains a list of databses maintained or proposed by those who replied to a questionnaire. (author)

Complexes of γ-tubulin with nonreceptor protein tyrosine kinases Src and Fyn in differentiating P19 embryonal carcinoma cells

International Nuclear Information System (INIS)

Kukharskyy, Vitaliy; Sulimenko, Vadym; Macurek, Libor; Sulimenko, Tetyana; Draberova, Eduarda; Draber, Pavel

2004-01-01

Nonreceptor protein tyrosine kinases of the Src family have been shown to play an important role in signal transduction as well as in regulation of microtubule protein interactions. Here we show that γ-tubulin (γ-Tb) in P19 embryonal carcinoma cells undergoing neuronal differentiation is phosphorylated and forms complexes with protein tyrosine kinases of the Src family, Src and Fyn. Elevated expression of both kinases during differentiation corresponded with increased level of proteins phosphorylated on tyrosine. Immunoprecipitation experiments with antibodies against Src, Fyn, γ-tubulin, and with anti-phosphotyrosine antibody revealed that γ-tubulin appeared in complexes with these kinases. In vitro kinase assays showed tyrosine phosphorylation of proteins in γ-tubulin complexes isolated from differentiated cells. Pretreatment of cells with Src family selective tyrosine kinase inhibitor PP2 reduced the amount of phosphorylated γ-tubulin in the complexes. Binding experiments with recombinant SH2 and SH3 domains of Src and Fyn kinases revealed that protein complexes containing γ-tubulin bound to SH2 domains and that these interactions were of SH2-phosphotyrosine type. The combined data suggest that Src family kinases might have an important role in the regulation of γ-tubulin interaction with tubulin dimers or other proteins during neurogenesis

Ror2-Src signaling in metastasis of mouse melanoma cells is inhibited by NRAGE.

Science.gov (United States)

Lai, Shan-Shan; Xue, Bin; Yang, Yang; Zhao, Li; Chu, Chao-Shun; Hao, Jia-Yin; Wen, Chuan-Jun

2012-11-01

The receptor tyrosine kinase (RTK) Ror2 plays important roles in developmental morphogenesis and mediates the filopodia formation in Wnt5a-induced cell migration. However, the function of Ror2 in noncanonical Wnt signaling resulting in cancer metastasis is largely unknown. Here, we show that Ror2 expression is higher in the highly metastatic murine B16-BL6 melanoma cells than in the low metastatic variant B16 cells. Overexpression of Ror2 increases the metastasis ability of B16 cells, and knockdown of Ror2 reduces the migration ability of B16-BL6 cells. Furthermore, the inhibition of Src kinase activity is critical for the Ror2-mediated cell migration upon Wnt5a treatment. The C-terminus of Ror2, which is deleted in brachydactyly type B (BDB), is essential for the mutual interaction with the SH1 domain of Src. Intriguingly, the Neurotrophin receptor-interacting MAGE homologue (NRAGE), which, as we previously reported, can remodel the cellular skeleton and inhibit cell-cell adhesion and metastasis of melanoma and pancreatic cancer, sharply blocks the interaction between Src and Ror2 and inhibits Ror2-mediated B16 cell migration by decreasing the activity of Src and focal adhesion kinase (FAK). Our data show that Ror2 is a potential factor in the tumorigenesis and metastasis in a Src-dependent manner that is negatively regulated by NRAGE. Copyright © 2012. Published by Elsevier Inc.

v-Src oncogene product increases sphingosine kinase 1 expression through mRNA stabilization: alteration of AU-rich element-binding proteins.

Science.gov (United States)

Sobue, S; Murakami, M; Banno, Y; Ito, H; Kimura, A; Gao, S; Furuhata, A; Takagi, A; Kojima, T; Suzuki, M; Nozawa, Y; Murate, T

2008-10-09

Sphingosine kinase 1 (SPHK1) is overexpressed in solid tumors and leukemia. However, the mechanism of SPHK1 overexpression by oncogenes has not been defined. We found that v-Src-transformed NIH3T3 cells showed a high SPHK1 mRNA, SPHK1 protein and SPHK enzyme activity. siRNA of SPHK1 inhibited the growth of v-Src-NIH3T3, suggesting the involvement of SPHK1 in v-Src-induced oncogenesis. v-Src-NIH3T3 showed activations of protein kinase C-alpha, signal transducers and activators of transcription 3 and c-Jun NH(2)-terminal kinase. Their inhibition suppressed SPHK1 expression in v-Src-NIH3T3, whereas their overexpression increased SPHK1 mRNA in NIH3T3. Unexpectedly, the nuclear run-on assay and the promoter analysis using 5'-promoter region of mouse SPHK1 did not show any significant difference between mock- and v-Src-NIH3T3. Furthermore, the half-life of SPHK1 mRNA in mock-NIH3T3 was nearly 15 min, whereas that of v-Src-NIH3T3 was much longer. Examination of two AU-rich region-binding proteins, AUF1 and HuR, that regulate mRNA decay reciprocally, showed decreased total AUF1 protein associated with increased tyrosine-phosphorylated form and increased serine-phosphorylated HuR protein in v-Src-NIH3T3. Modulation of AUF1 and HuR by their overexpression or siRNA revealed that SPHK1 mRNA in v-Src- and mock-NIH3T3 was regulated reciprocally by these factors. Our results showed, for the first time, a novel mechanism of v-Src-induced SPHK1 overexpression.

Flow-Based Provenance

Directory of Open Access Journals (Sweden)

Sabah Al-Fedaghi

2017-02-01

Full Text Available Aim/Purpose: With information almost effortlessly created and spontaneously available, current progress in Information and Communication Technology (ICT has led to the complication that information must be scrutinized for trustworthiness and provenance. Information systems must become provenance-aware to be satisfactory in accountability, reproducibility, and trustworthiness of data. Background:\tMultiple models for abstract representation of provenance have been proposed to describe entities, people, and activities involved in producing a piece of data, including the Open Provenance Model (OPM and the World Wide Web Consortium. These models lack certain concepts necessary for specifying workflows and encoding the provenance of data products used and generated. Methodology: Without loss of generality, the focus of this paper is on OPM depiction of provenance in terms of a directed graph. We have redrawn several case studies in the framework of our proposed model in order to compare and evaluate it against OPM for representing these cases. Contribution: This paper offers an alternative flow-based diagrammatic language that can form a foundation for modeling of provenance. The model described here provides an (abstract machine-like representation of provenance. Findings: The results suggest a viable alternative in the area of diagrammatic representation for provenance applications. Future Research: Future work will seek to achieve more accurate comparisons with current models in the field.

Selective Targeting of SH2 Domain-Phosphotyrosine Interactions of Src Family Tyrosine Kinases with Monobodies.

Science.gov (United States)

Kükenshöner, Tim; Schmit, Nadine Eliane; Bouda, Emilie; Sha, Fern; Pojer, Florence; Koide, Akiko; Seeliger, Markus; Koide, Shohei; Hantschel, Oliver

2017-05-05

The binding of Src-homology 2 (SH2) domains to phosphotyrosine (pY) sites is critical for the autoinhibition and substrate recognition of the eight Src family kinases (SFKs). The high sequence conservation of the 120 human SH2 domains poses a significant challenge to selectively perturb the interactions of even the SFK SH2 family against the rest of the SH2 domains. We have developed synthetic binding proteins, termed monobodies, for six of the SFK SH2 domains with nanomolar affinity. Most of these monobodies competed with pY ligand binding and showed strong selectivity for either the SrcA (Yes, Src, Fyn, Fgr) or SrcB subgroup (Lck, Lyn, Blk, Hck). Interactome analysis of intracellularly expressed monobodies revealed that they bind SFKs but no other SH2-containing proteins. Three crystal structures of monobody-SH2 complexes unveiled different and only partly overlapping binding modes, which rationalized the observed selectivity and enabled structure-based mutagenesis to modulate inhibition mode and selectivity. In line with the critical roles of SFK SH2 domains in kinase autoinhibition and T-cell receptor signaling, monobodies binding the Src and Hck SH2 domains selectively activated respective recombinant kinases, whereas an Lck SH2-binding monobody inhibited proximal signaling events downstream of the T-cell receptor complex. Our results show that SFK SH2 domains can be targeted with unprecedented potency and selectivity using monobodies. They are excellent tools for dissecting SFK functions in normal development and signaling and to interfere with aberrant SFK signaling networks in cancer cells. Copyright © 2017 The Author(s). Published by Elsevier Ltd.. All rights reserved.

Src is activated by the nuclear receptor peroxisome proliferator-activated receptor β/δ in ultraviolet radiation-induced skin cancer.

Science.gov (United States)

Montagner, Alexandra; Delgado, Maria B; Tallichet-Blanc, Corinne; Chan, Jeremy S K; Sng, Ming K; Mottaz, Hélén; Degueurce, Gwendoline; Lippi, Yannick; Moret, Catherine; Baruchet, Michael; Antsiferova, Maria; Werner, Sabine; Hohl, Daniel; Saati, Talal Al; Farmer, Pierre J; Tan, Nguan S; Michalik, Liliane; Wahli, Walter

2014-01-01

Although non-melanoma skin cancer (NMSC) is the most common human cancer and its incidence continues to rise worldwide, the mechanisms underlying its development remain incompletely understood. Here, we unveil a cascade of events involving peroxisome proliferator-activated receptor (PPAR) β/δ and the oncogene Src, which promotes the development of ultraviolet (UV)-induced skin cancer in mice. UV-induced PPARβ/δ activity, which directly stimulated Src expression, increased Src kinase activity and enhanced the EGFR/Erk1/2 signalling pathway, resulting in increased epithelial-to-mesenchymal transition (EMT) marker expression. Consistent with these observations, PPARβ/δ-null mice developed fewer and smaller skin tumours, and a PPARβ/δ antagonist prevented UV-dependent Src stimulation. Furthermore, the expression of PPARβ/δ positively correlated with the expression of SRC and EMT markers in human skin squamous cell carcinoma (SCC), and critically, linear models applied to several human epithelial cancers revealed an interaction between PPARβ/δ and SRC and TGFβ1 transcriptional levels. Taken together, these observations motivate the future evaluation of PPARβ/δ modulators to attenuate the development of several epithelial cancers.

A role for Pyk2 and Src in linking G-protein-coupled receptors with MAP kinase activation.

Science.gov (United States)

Dikic, I; Tokiwa, G; Lev, S; Courtneidge, S A; Schlessinger, J

1996-10-10

The mechanisms by which mitogenic G-protein-coupled receptors activate the MAP kinase signalling pathway are poorly understood. Candidate protein tyrosine kinases that link G-protein-coupled receptors with MAP kinase include Src family kinases, the epidermal growth factor receptor, Lyn and Syk. Here we show that lysophosphatidic acid (LPA) and bradykinin induce tyrosine phosphorylation of Pyk2 and complex formation between Pyk2 and activated Src. Moreover, tyrosine phosphorylation of Pyk2 leads to binding of the SH2 domain of Src to tyrosine 402 of Pyk2 and activation of Src. Transient overexpression of a dominant interfering mutant of Pyk2 or the protein tyrosine kinase Csk reduces LPA- or bradykinin-induced activation of MAP kinase. LPA- or bradykinin-induced MAP kinase activation was also inhibited by overexpression of dominant interfering mutants of Grb2 and Sos. We propose that Pyk2 acts with Src to link Gi- and Gq-coupled receptors with Grb2 and Sos to activate the MAP kinase signalling pathway in PC12 cells.

Interaction with the Src homology (SH3-SH2) region of the Src-family kinase Hck structures the HIV-1 Nef dimer for kinase activation and effector recruitment.

Science.gov (United States)

Alvarado, John Jeff; Tarafdar, Sreya; Yeh, Joanne I; Smithgall, Thomas E

2014-10-10

HIV-1 Nef supports high titer viral replication in vivo and is essential for AIDS progression. Nef function depends on interactions with multiple host cell effectors, including Hck and other Src-family kinases. Here we describe the x-ray crystal structure of Nef in complex with the Hck SH3-SH2 regulatory region to a resolution of 1.86 Å. The complex crystallized as a dimer of complexes, with the conserved Nef PXXPXR motif engaging the Hck SH3 domain. A new intercomplex contact was found between SH3 Glu-93, and Nef Arg-105. Mutagenesis of Hck SH3 Glu-93 interfered with Nef·Hck complex formation and kinase activation in cells. The Hck SH2 domains impinge on the N-terminal region of Nef to stabilize a dimer conformation that exposes Asp-123, a residue critical for Nef function. Our results suggest that in addition to serving as a kinase effector for Nef, Hck binding may reorganize the Nef dimer for functional interaction with other signaling partners. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

Interaction with the Src Homology (SH3-SH2) Region of the Src-family Kinase Hck Structures the HIV-1 Nef Dimer for Kinase Activation and Effector Recruitment*

Science.gov (United States)

Alvarado, John Jeff; Tarafdar, Sreya; Yeh, Joanne I.; Smithgall, Thomas E.

2014-01-01

HIV-1 Nef supports high titer viral replication in vivo and is essential for AIDS progression. Nef function depends on interactions with multiple host cell effectors, including Hck and other Src-family kinases. Here we describe the x-ray crystal structure of Nef in complex with the Hck SH3-SH2 regulatory region to a resolution of 1.86 Å. The complex crystallized as a dimer of complexes, with the conserved Nef PXXPXR motif engaging the Hck SH3 domain. A new intercomplex contact was found between SH3 Glu-93, and Nef Arg-105. Mutagenesis of Hck SH3 Glu-93 interfered with Nef·Hck complex formation and kinase activation in cells. The Hck SH2 domains impinge on the N-terminal region of Nef to stabilize a dimer conformation that exposes Asp-123, a residue critical for Nef function. Our results suggest that in addition to serving as a kinase effector for Nef, Hck binding may reorganize the Nef dimer for functional interaction with other signaling partners. PMID:25122770

v-src induction of the TIS10/PGS2 prostaglandin synthase gene is mediated by an ATF/CRE transcription response element.

Science.gov (United States)

Xie, W; Fletcher, B S; Andersen, R D; Herschman, H R

1994-10-01

We recently reported the cloning of a mitogen-inducible prostaglandin synthase gene, TIS10/PGS2. In addition to growth factors and tumor promoters, the v-src oncogene induces TIS10/PGS2 expression in 3T3 cells. Deletion analysis, using luciferase reporters, identifies a region between -80 and -40 nucleotides 5' of the TIS10/PGS2 transcription start site that mediates pp60v-src induction in 3T3 cells. This region contains the sequence CGTCACGTG, which includes overlapping ATF/CRE (CGTCA) and E-box (CACGTG) sequences. Gel shift-oligonucleotide competition experiments with nuclear extracts from cells stably transfected with a temperature-sensitive v-src gene demonstrate that the CGTCACGTG sequence can bind proteins at both the ATF/CRE and E-box sequences. Dominant-negative CREB and Myc proteins that bind DNA, but do not transactivate, block v-src induction of a luciferase reporter driven by the first 80 nucleotides of the TIS10/PGS2 promoter. Mutational analysis distinguishes which TIS10/PGS2 cis-acting element mediates pp60v-src induction. E-box mutation has no effect on the fold induction in response to pp60v-src. In contrast, ATF/CRE mutation attenuates the pp60v-src response. Antibody supershift and methylation interference experiments demonstrate that CREB and at least one other ATF transcription factor in these extracts bind to the TIS10/PGS2 ATF/CRE element. Expression of a dominant-negative ras gene also blocks TIS10/PGS2 induction by v-src. Our data suggest that Ras mediates pp60v-src activation of an ATF transcription factor, leading to induced TIS10/PGS2 expression via the ATF/CRE element of the TIS10/PGS2 promoter. This is the first description of v-src activation of gene expression via an ATF/CRE element.

Isthmin is a novel vascular permeability inducer that functions through cell-surface GRP78-mediated Src activation.

Science.gov (United States)

Venugopal, Shruthi; Chen, Mo; Liao, Wupeng; Er, Shi Yin; Wong, Wai-Shiu Fred; Ge, Ruowen

2015-07-01

Isthmin (ISM) is a recently identified 60 kDa secreted angiogenesis inhibitor. Two cell-surface receptors for ISM have been defined, the high-affinity glucose-regulated protein 78 kDa (GRP78) and the low-affinity αvβ5 integrin. As αvβ5 integrin plays an important role in pulmonary vascular permeability (VP) and ISM is highly expressed in mouse lung, we sought to clarify the role of ISM in VP. Recombinant ISM (rISM) dose-dependently enhances endothelial monolayer permeability in vitro and local dermal VP when administered intradermally in mice. Systemic rISM administration through intravenous injection leads to profound lung vascular hyperpermeability but not in other organs. Mechanistic investigations using molecular, biochemical approaches and specific chemical inhibitors revealed that ISM-GRP78 interaction triggers a direct interaction between GRP78 and Src, leading to Src activation and subsequent phosphorylation of adherens junction proteins and loss of junctional proteins from inter-endothelial junctions, resulting in enhanced VP. Dynamic studies of Src activation, VP and apoptosis revealed that ISM induces VP directly via Src activation while apoptosis contributes indirectly only after prolonged treatment. Furthermore, ISM is significantly up-regulated in lipopolysaccharide (LPS)-treated mouse lung. Blocking cell-surface GRP78 by systemic infusion of anti-GRP78 antibody significantly attenuates pulmonary vascular hyperpermeability in LPS-induced acute lung injury (ALI) in mice. ISM is a novel VP inducer that functions through cell-surface GRP78-mediated Src activation as well as induction of apoptosis. It induces a direct GRP78-Src interaction, leading to cytoplasmic Src activation. ISM contributes to pulmonary vascular hyperpermeability of LPS-induced ALI in mice. Published on behalf of the European Society of Cardiology. All rights reserved. © The Author 2015. For permissions please email: journals.permissions@oup.com.

Src kinases regulate de novo actin polymerization during exocytosis in neuroendocrine chromaffin cells.

Directory of Open Access Journals (Sweden)

María José Olivares

Full Text Available The cortical actin network is dynamically rearranged during secretory processes. Nevertheless, it is unclear how de novo actin polymerization and the disruption of the preexisting actin network control transmitter release. Here we show that in bovine adrenal chromaffin cells, both formation of new actin filaments and disruption of the preexisting cortical actin network are induced by Ca2+ concentrations that trigger exocytosis. These two processes appear to regulate different stages of exocytosis; whereas the inhibition of actin polymerization with the N-WASP inhibitor wiskostatin restricts fusion pore expansion, thus limiting the release of transmitters, the disruption of the cortical actin network with cytochalasin D increases the amount of transmitter released per event. Further, the Src kinase inhibitor PP2, and cSrc SH2 and SH3 domains also suppress Ca2+-dependent actin polymerization, and slow down fusion pore expansion without disturbing the cortical F-actin organization. Finally, the isolated SH3 domain of c-Src prevents both the disruption of the actin network and the increase in the quantal release induced by cytochalasin D. These findings support a model where a rise in the cytosolic Ca2+ triggers actin polymerization through a mechanism that involves Src kinases. The newly formed actin filaments would speed up the expansion of the initial fusion pore, whereas the preexisting actin network might control a different step of the exocytosis process.

Melatonin regulates CRE-dependent gene transcription underlying osteoblast proliferation by activating Src and PKA in parallel.

Science.gov (United States)

Tao, Lin; Zhu, Yue

2018-01-01

Several studies have indicated a relationship between melatonin and idiopathic scoliosis, including our previous work which demonstrated that melatonin can inhibit osteoblast proliferation; however, the mechanism remains unclear. Here, we utilized a MTT assay to show that melatonin significantly reduces osteoblast proliferation in a concentration-and time-dependent manner. Through a combination of techniques, including real-time PCR, MTT assays, immunofluorescence, and luciferase assays, we confirmed that melatonin-induced changes in phosphorylated cAMP response element-binding protein (CREB) reduced transcriptional activity in a melatonin receptor-dependent manner. Surprisingly, treatment of osteoblasts with the mitogen-activated protein kinase/extracellular signal-regulated kinase kinase (MEK) inhibitor PD98059 up-regulated other cascades upstream of CREB. We next treated cells with PKA and Src inhibitors and observed that melatonin can also activate the protein kinase A (PKA) and Src pathways. To examine whether Src is upstream from the cAMP-PKA pathway, we measured cAMP levels in response to melatonin with and without a Src inhibitor (PP2) and found that PP2 had no additional effect. Therefore, the transcription-dependent mechanisms involved in CREB phosphorylation, along with melatonin, activated Src via a parallel signaling pathway that was separate from that of PKA. Finally, we transfected osteoblasts with lentiviral CREB short hairpin (sh) RNAs and found a decrease in the expression of proliferating cell nuclear antigen (PCNA) and osteoblast proliferation. These results suggest that CREB and PCNA are downstream targets of melatonin signaling, and that the down-regulation of CREB, which is regulated via PKA and Src pathways, contributes to the melatonin-induced inhibition of osteoblast proliferation.

A Discovery Strategy for Selective Inhibitors of c-Src in Complex with the Focal Adhesion Kinase SH3/SH2-binding Region.

Science.gov (United States)

Moroco, Jamie A; Baumgartner, Matthew P; Rust, Heather L; Choi, Hwan Geun; Hur, Wooyoung; Gray, Nathanael S; Camacho, Carlos J; Smithgall, Thomas E

2015-08-01

The c-Src tyrosine kinase co-operates with the focal adhesion kinase to regulate cell adhesion and motility. Focal adhesion kinase engages the regulatory SH3 and SH2 domains of c-Src, resulting in localized kinase activation that contributes to tumor cell metastasis. Using assay conditions where c-Src kinase activity required binding to a tyrosine phosphopeptide based on the focal adhesion kinase SH3-SH2 docking sequence, we screened a kinase-biased library for selective inhibitors of the Src/focal adhesion kinase peptide complex versus c-Src alone. This approach identified an aminopyrimidinyl carbamate compound, WH-4-124-2, with nanomolar inhibitory potency and fivefold selectivity for c-Src when bound to the phospho-focal adhesion kinase peptide. Molecular docking studies indicate that WH-4-124-2 may preferentially inhibit the 'DFG-out' conformation of the kinase active site. These findings suggest that interaction of c-Src with focal adhesion kinase induces a unique kinase domain conformation amenable to selective inhibition. © 2014 John Wiley & Sons A/S.

Harvester development for new high yielding SRC crops and markets

Energy Technology Data Exchange (ETDEWEB)

Paulson, Mark

2005-07-01

This report describes the development of harvesting equipment for short rotation cultivation (SRC) crops produced in the UK that can produce fuel to a required specification in a single pass at a cost that is profitable for the grower while minimising the cost of the product. Details are given of the manufacture and installation of new components for large crop harvesting, and production of fuel suitable for co-firing in a coal combustion system using pulverised fuel and fuel suitable for gasification. The development of the drive chain to cope with the higher yielding crops, field tests on SRC crops, and determination of the most economic harvesting system are discussed along with the remanufacture of the chipping drum, and production of market chip samples. Harvesting guidance and an economic analysis of harvesting systems are presented.

Interplay of Matrix Stiffness and c-SRC in Hepatic Fibrosis

DEFF Research Database (Denmark)

Görtzen, Jan; Schierwagen, Robert; Bierwolf, Jeanette

2015-01-01

. This study investigated the interaction of c-SRC and RhoA under different matrix stiffness conditions. METHODS: Liver fibrosis was induced in rats using bile duct ligation (BDL), thioacetamide (TAA) or carbon tetrachloride (CCl4) models. mRNA levels of albumin, PDGF-R, RHOA, COL1A1, and αSMA were analyzed......INTRODUCTION: In liver fibrosis activation of hepatic stellate cells (HSC) comprises phenotypical change into profibrotic and myofibroplastic cells with increased contraction and secretion of extracellular matrix (ECM) proteins. The small GTPase RhoA orchestrates cytoskeleton formation, migration......, and mobility via non-receptor tyrosine-protein kinase c-SRC (cellular sarcoma) in different cells. Furthermore, RhoA and its downstream effector Rho-kinase also play a crucial role in hepatic stellate cells and hepatic fibrogenesis. Matrix stiffness promotes HSC activation via cytoskeleton modulation...

Harvester development for new high yielding SRC crops and markets

International Nuclear Information System (INIS)

Paulson, Mark

2005-01-01

This report describes the development of harvesting equipment for short rotation cultivation (SRC) crops produced in the UK that can produce fuel to a required specification in a single pass at a cost that is profitable for the grower while minimising the cost of the product. Details are given of the manufacture and installation of new components for large crop harvesting, and production of fuel suitable for co-firing in a coal combustion system using pulverised fuel and fuel suitable for gasification. The development of the drive chain to cope with the higher yielding crops, field tests on SRC crops, and determination of the most economic harvesting system are discussed along with the remanufacture of the chipping drum, and production of market chip samples. Harvesting guidance and an economic analysis of harvesting systems are presented

1-o-acetylbritannilactone (ABL) inhibits angiogenesis and lung cancer cell growth through regulating VEGF-Src-FAK signaling

Energy Technology Data Exchange (ETDEWEB)

Zhengfu, He; Hu, Zhang; Huiwen, Miao; Zhijun, Li [Department of Thoracic Surgery, Sir Run Run Shaw Hospital of Zhejiang University School of Medicine, Hangzhou (China); Jiaojie, Zhou [Zhejiang University School of Medicine, Hangzhou (China); Xiaoyi, Yan, E-mail: xiaoyiyan163@163.com [Zhejiang University School of Medicine, Hangzhou (China); Xiujun, Cai, E-mail: xiujuncaomaj@163.com [Sir Run Run Shaw Hospital of Zhejiang University School of Medicine, Hangzhou (China)

2015-08-21

The search for safe, effective and affordable therapeutics against non-small cell lung cancer (NSCLC) and other lung cancers is important. Here we explored the potential effect of 1-o-acetylbritannilactone (ABL), a novel extract from Inula britannica-F, on angiogenesis and lung cancer cell growth. We demonstrated that ABL dose-dependently inhibited vascular endothelial growth factor (VEGF)-induced proliferation, migration, and capillary structure formation of cultured human umbilical vascular endothelial cells (HUVECs). In vivo, ABL administration suppressed VEGF-induced new vasculature formation in Matrigel plugs. For the mechanism investigations, we found that ABL largely inhibited VEGF-mediated activation of Src kinase and focal adhesion kinase (FAK) in HUVECs. Furthermore, treatment of A549 NSCLC cells with ABL resulted in cell growth inhibition and Src-FAK in-activation. Significantly, administration of a single dose of ABL (12 mg/kg/day) remarkably suppressed growth of A549 xenografts in nude mice. In vivo microvessels formation and Src activation were also significantly inhibited in ABL-treated xenograft tumors. Taken together, our findings suggest that ABL suppresses angiogenesis and lung cancer cell growth possibly via regulating the VEGFR-Src-FAK signaling. - Highlights: • 1-o-acetylbritannilactone (ABL) inhibits VEGF-induced angiogenesis in vivo. • ABL inhibits VEGF-induced HUVEC migration, proliferation, capillary tube formation. • ABL inhibits VEGF-mediated activation of Src and FAK in HUVECs. • ABL inhibits growth and Src-FAK activation in A549 cells. • ABL administration inhibits A549 tumor angiogenesis and growth in nude mice.

v-src Induction of the TIS10/PGS2 prostaglandin synthase gene is mediated by an ATF/CRE transcription response element

Energy Technology Data Exchange (ETDEWEB)

Xie, W.; Fletcher, B.S.; Andersen, R.D.; Herschman, H.R. [Univ. of California, Los Angeles, CA (United States)

1994-10-01

The authors recently reported the cloning of a mitogen-inducible prostaglandin synthase gene, TIS10/PGS2. In addition to growth factor and tumor promoters, the v-src oncogene induces TIS10/PGS2 expression in 3T3 cells. Deletion analysis, using luciferase reporters, identifies a region between -80 and -40 nucleotides 5{prime} of the TIS10/PGS2 transcription start site that mediates pp60{sup v-src} induction in 3T3 cells. This region contains the sequence CGTCACGTG, which includes overlapping ATF/CRE (CGTCA) and E-box (CACGTG) sequences. Gel shift-oligonucleotide competition experiments with nuclear extracts from cells stably transfected with a temperature-sensitive v-src gene demonstrate that the CGTCACGTG sequence can bind proteins at both the AFT/CRE and E-box sequences. Dominant-negative CREB and Myc proteins that bind DNA, but do not transactivate, block v-src induction of a luciferase reporter driven by the first 80 nucleotides of the TIS10/PGS2 promoter. Mutational analysis distinguishes which TIS10/PGS2 cis-acting element mediates pp60{sup v-src} induction. E-box mutation has no effect on the fold induction in response to pp60{sup v-src}. In contrast, ATF/CRE mutation attenuates the pp{sup v-src} response. Antibody supershift and methylation interference experiments demonstrate that CREB and at least one other ATF transcription factor in these extracts bind to the TIS10/PGS2 ATF/CRE element. Expression of a dominant-negative ras gene also blocks TIS10/PGS2 induction by v-src. The data suggest that Ras mediates pp60{sup v-src} activation of an ATF transcription factor, leading to induced TIS10/PGS2 expression via the ATF/CRE element of the TIS10/PGS2 promoter. This is the first description of v-src activation of gene expression via an ATF/CRE element. 64 refs., 8 figs.

Band 3 tyrosine kinase in avian erythrocyte plasma membrane is immunologically related to pp60c-src

International Nuclear Information System (INIS)

Hillsgrove, D.; Shores, C.G.; Parker, J.C.; Maness, P.F.

1987-01-01

The authors have identified in the plasma membrane of the chicken erythrocyte a 60-kDa tyrosine-specific protein kinase immunologically related to the transforming protein pp60 v-src of Rous sarcoma virus. The erythrocyte protein kinase phosphorylated heavy chains of tumor-bearing rabbit (TBR) antibodies reactive with pp60 c-src at tyrosine in immune complex protein kinase assays. The kinase was identified as a 60-kDa protein by [ 35 S]methionine labeling of erythrocytes and by autophosphorylation in immune complexes. The kinase migrated on two-dimensional gel electrophoresis with an apparent pI and molecular mass similar to pp60 c-src . A plasma membrane-enriched fraction isolated from chicken red cells contained the majority of the kinase activity. Incubation of the plasma membrane fraction with [ 32 P]ATP resulted in tyrosine phosphorylation of the anion transport protein band 3. Band 3 phosphorylation was blocked by TBR antibodies, indicting that the kinase recognized by pp60 c-src antibodies was responsible for band 3 phosphorylation. These results demonstrate that the avian erythrocyte plasma membrane contains a tightly bound tyrosine-specific protein kinase identical or closely related to pp60 c-src and that this kinase is responsible for band 3 phosphorylation in vitro

The v-Src and c-Src tyrosine kinases immunoprecipitated from Rous sarcoma virus-transformed cells display different peptide substrate specificities

Czech Academy of Sciences Publication Activity Database

Vojtěchová, Martina; Tuháčková, Zdena; Hlaváček, Jan; Velek, Jiří; Sovová, Vlasta

2004-01-01

Roč. 421, č. 2 (2004), s. 277-282 ISSN 0003-9861 R&D Projects: GA ČR GV312/96/K205; GA ČR GA301/00/0269 Institutional research plan: CEZ:AV0Z4055905; CEZ:AV0Z1003909 Keywords : Src kinase, in vitro phosphorylation, peptide substrate specificity Subject RIV: CE - Biochemistry Impact factor: 2.657, year: 2004

Micro-Environmental Stress Induces Src-Dependent Activation of Invadopodia and Cell Migration in Ewing Sarcoma

Directory of Open Access Journals (Sweden)

Kelly M. Bailey

2016-08-01

Full Text Available Metastatic Ewing sarcoma has a very poor prognosis and therefore new investigations into the biologic drivers of metastatic progression are key to finding new therapeutic approaches. The tumor microenvironment is highly dynamic, leading to exposure of different regions of a growing solid tumor to changes in oxygen and nutrient availability. Tumor cells must adapt to such stress in order to survive and propagate. In the current study, we investigate how Ewing sarcoma cells respond to the stress of growth factor deprivation and hypoxia. Our findings reveal that serum deprivation leads to a reversible change in Ewing cell cytoskeletal phenotypes. Using an array of migration and invasion techniques, including gelatin matrix degradation invadopodia assays, we show that exposure of Ewing sarcoma cells to serum deprivation and hypoxia triggers enhanced migration, invadopodia formation, matrix degradation and invasion. Further, these functional changes are accompanied by and dependent on activation of Src kinase. Activation of Src, and the associated invasive cell phenotype, were blocked by exposing hypoxia and serum-deprived cells to the Src inhibitor dasatinib. These results indicate that Ewing sarcoma cells demonstrate significant plasticity in response to rapidly changing micro-environmental stresses that can result from rapid tumor growth and from necrosis-causing therapies. In response to these stresses, Ewing cells transition to a more migratory and invasive state and our data show that Src is an important mediator of this stress response. Our data support exploration of clinically available Src inhibitors as adjuvant agents for metastasis prevention in Ewing sarcoma.

76 FR 1458 - Public Meeting for the National Park Service Alaska Region's Subsistence Resource Commission (SRC...

Science.gov (United States)

2011-01-10

... Plan Update. c. Subsistence Uses of Horns, Antlers, Bones and Plants EA Update. 13. New Business. 14... guarantee that we will be able to do so. Wrangell-St. Elias National Park SRC Meeting Date and Location: The... if all business is completed. For Further Information on the Gates of the Arctic National Park SRC...

Alterations in c-Src/HER1 and estrogen receptor α signaling pathways in mammary gland and tumors of hexachlorobenzene-treated rats

International Nuclear Information System (INIS)

Peña, Delfina; Pontillo, Carolina; García, María Alejandra; Cocca, Claudia; Alvarez, Laura; Chiappini, Florencia; Bourguignon, Nadia; Frahm, Isabel; Bergoc, Rosa; Kleiman de Pisarev, Diana; Randi, Andrea

2012-01-01

Hexachlorobenzene (HCB) is an organochlorine pesticide that acts as an endocrine disruptor in humans and rodents. The development of breast cancer strongly depends on endocrine conditions modulated by environmental factors. We have demonstrated that HCB is a tumor co-carcinogen in rats and an inducer of proliferation in MCF-7 cells, in an estrogen receptor α (ERα)-dependent manner, and of migration in MDA-MB-231 breast cancer cell line. In the present study, we examined HCB effect on c-Src/human epidermal growth factor receptor (HER1) and ERα signaling pathways in mammary glands and in N-nitroso-N-methylurea (NMU)-induced mammary tumors in rats. Furthermore, we evaluated histopathological changes and serum hormone levels. Rats were separated into four groups: control, HCB (100 mg/kg b.w.), NMU (50 mg/kg b.w.) and NMU-HCB. Our data show that HCB increases c-Src and HER1 activation, c-Src/HER1 association, and Y699-STAT5b and ERK1/2 phosphorylation in mammary glands. HCB also enhances Y537-ERα phosphorylation and ERα/c-Src physical interaction. In tumors, HCB also induces c-Src and HER1 activation, c-Src/HER1 association, as well as T308-Akt and Y699-STAT5b phosphorylation. In addition, the pesticide increases ERα protein content and decreases p-Y537-ERα levels and ERα/c-Src association in tumors. HCB increases serum 17-beta estradiol and prolactin contents and decreases progesterone, FSH and LH levels in rats without tumors, while the opposite effect was observed in rats with tumors. Taken together, our results indicate that HCB induces an estrogenic effect in mammary gland, increasing c-Src/HER1 and ERα signaling pathways. HCB stimulates c-Src/HER1 pathway, but decreases ERα activity in tumors, appearing to shift them towards a higher malignancy phenotype.

Csk-Induced Phosphorylation of Src at Tyrosine 530 is Essential for H2O2-Mediated Suppression of ERK1/2 in Human Umbilical Vein Endothelial Cells

Science.gov (United States)

Jeon, Bo Kyung; Kwon, Kihwan; Kang, Jihee Lee; Choi, Youn-Hee

2015-01-01

Mitogen-activated protein kinases (MAPKs) are key signal transducers involved in various cellular events such as growth, proliferation, and differentiation. Previous studies have reported that H2O2 leads to phosphorylation of extracellular signal-regulated kinase (ERK), one of the MAPKs in endothelial cells. The current study shows that H2O2 suppressed ERK1/2 activation and phosphorylation at specific concentrations and times in human umbilical vein endothelial cells but not in immortalized mouse aortic endothelial cells or human astrocytoma cell line CRT-MG. Phosphorylation of other MAPK family members (i.e., p38 and JNK) was not suppressed by H2O2. The decrease in ERK1/2 phosphorylation induced by H2O2 was inversely correlated with the level of phosphorylation of Src tyrosine 530. Using siRNA, it was found that H2O2-induced suppression of ERK1/2 was dependent on Csk. Physiological laminar flow abrogated, but oscillatory flow did not affect, the H2O2-induced suppression of ERK1/2 phosphorylation. In conclusion, H2O2-induced Csk translocation to the plasma membrane leads to phosphorylation of Src at the tyrosine 530 residue resulting in a reduction of ERK1/2 phosphorylation. Physiological laminar flow abrogates this effect of H2O2 by inducing phosphorylation of Src tyrosine 419. These findings broaden our understanding of signal transduction mechanisms in the endothelial cells against oxidative stress. PMID:26234813

Abnormal Cell Properties and Down-Regulated FAK-Src Complex Signaling in B Lymphoblasts of Autistic Subjects

Science.gov (United States)

Wei, Hongen; Malik, Mazhar; Sheikh, Ashfaq M.; Merz, George; Ted Brown, W.; Li, Xiaohong

2011-01-01

Recent studies suggest that one of the major pathways to the pathogenesis of autism is reduced cell migration. Focal adhesion kinase (FAK) has an important role in neural migration, dendritic morphological characteristics, axonal branching, and synapse formation. The FAK-Src complex, activated by upstream reelin and integrin β1, can initiate a cascade of phosphorylation events to trigger multiple intracellular pathways, including mitogen-activated protein kinase–extracellular signal–regulated kinase and phosphatidylinositol 3-kinase–Akt signaling. In this study, by using B lymphoblasts as a model, we tested whether integrin β1 and FAK-Src signaling are abnormally regulated in autism and whether abnormal FAK-Src signaling leads to defects in B-lymphoblast adhesion, migration, proliferation, and IgG production. To our knowledge, for the first time, we show that protein expression levels of both integrin β1 and FAK are significantly decreased in autistic lymphoblasts and that Src protein expression and the phosphorylation of an active site (Y416) are also significantly decreased. We also found that lymphoblasts from autistic subjects exhibit significantly decreased migration, increased adhesion properties, and an impaired capacity for IgG production. The overexpression of FAK in autistic lymphoblasts countered the adhesion and migration defects. In addition, we demonstrate that FAK mediates its effect through the activation of Src, phosphatidylinositol 3-kinase–Akt, and mitogen-activated protein kinase signaling cascades and that paxillin is also likely involved in the regulation of adhesion and migration in autistic lymphoblasts. PMID:21703394

On root class residuality of HNN-extensions

International Nuclear Information System (INIS)

Tieudjo, D.

2004-08-01

A sufficient condition or root-class residuality of HNN-extensions with root-class residual base group is proven; namely if G = -1 1Ht = K, φ> is the HNN-extension with base group A, stable letter t and associated subgroups H and K via the isomorphism φ, then G is root-class residual if group A is root-class residual and there exists a homomorphism σ of group G onto some group of a root-class such that σ is one-to-one on H. For the particular case when H = K and σ is the identical map, it is shown that G is root-class residual if and only if A is root-class residual and subgroup H of A is root-class separable. These results are generalized to multiple HNN-extensions. (author)

Andrographolide downregulates the v-Src and Bcr-Abl oncoproteins and induces Hsp90 cleavage in the ROS-dependent suppression of cancer malignancy.

Science.gov (United States)

Liu, Sheng-Hung; Lin, Chao-Hsiung; Liang, Fong-Ping; Chen, Pei-Fen; Kuo, Cheng-Deng; Alam, Mohd Mujahid; Maiti, Barnali; Hung, Shih-Kai; Chi, Chin-Wen; Sun, Chung-Ming; Fu, Shu-Ling

2014-01-15

Andrographolide is a diterpenoid compound isolated from Andrographis paniculata that exhibits anticancer activity. We previously reported that andrographolide suppressed v-Src-mediated cellular transformation by promoting the degradation of Src. In the present study, we demonstrated the involvement of Hsp90 in the andrographolide-mediated inhibition of Src oncogenic activity. Using a proteomics approach, a cleavage fragment of Hsp90α was identified in andrographolide-treated cells. The concentration- and time-dependent induction of Hsp90 cleavage that accompanied the reduction in Src was validated in RK3E cells transformed with either v-Src or a human truncated c-Src variant and treated with andrographolide. In cancer cells, the induction of Hsp90 cleavage by andrographolide and its structural derivatives correlated well with decreased Src levels, the suppression of transformation, and the induction of apoptosis. Moreover, the andrographolide-induced Hsp90 cleavage, Src degradation, inhibition of transformation, and induction of apoptosis were abolished by a ROS inhibitor, N-acetyl-cysteine. Notably, Hsp90 cleavage, decreased levels of Bcr-Abl (another known Hsp90 client protein), and the induction of apoptosis were also observed in human K562 leukemia cells treated with andrographolide or its active derivatives. Together, we demonstrated a novel mechanism by which andrographolide suppressed cancer malignancy that involved inhibiting Hsp90 function and reducing the levels of Hsp90 client proteins. Our results broaden the molecular basis of andrographolide-mediated anticancer activity. Copyright © 2013 Elsevier Inc. All rights reserved.

The N-terminus of survivin is a mitochondrial-targeting sequence and Src regulator

Science.gov (United States)

Dunajová, Lucia; Cash, Emily; Markus, Robert; Rochette, Sophie; Townley, Amelia R.

2016-01-01

ABSTRACT Survivin (also known as BIRC5) is a cancer-associated protein that exists in several locations in the cell. Its cytoplasmic residence in interphase cells is governed by CRM1 (also known as XPO1)-mediated nuclear exportation, and its localisation during mitosis to the centromeres and midzone microtubules is that of a canonical chromosomal passenger protein. In addition to these well-established locations, survivin is also a mitochondrial protein, but how it gets there and its function therein is presently unclear. Here, we show that the first ten amino acids at the N-terminus of survivin are sufficient to target GFP to the mitochondria in vivo, and ectopic expression of this decapeptide decreases cell adhesion and accelerates proliferation. The data support a signalling mechanism in which this decapeptide regulates the tyrosine kinase Src, leading to reduced focal adhesion plaques and disruption of F-actin organisation. This strongly suggests that the N-terminus of survivin is a mitochondrial-targeting sequence that regulates Src, and that survivin acts in concert with Src to promote tumorigenesis. PMID:27246243

1170-MW(t) HTGR-PS/C plant application study report: SRC-II process application

International Nuclear Information System (INIS)

Rao, R.; McMain, A.T. Jr.

1981-05-01

The solvent refined coal (SRC-II) process is an advanced process being developed by Gulf Mineral Resources Ltd. (a Gulf Oil Corporation subsidiary) to produce a clean, non-polluting liquid fuel from high-sulfur bituminous coals. The SRC-II commercial plant will process about 24,300 tonnes (26,800 tons) of feed coal per stream day, producing primarily fuel oil plus secondary fuel gases. This summary report describes the integration of a high-temperature gas-cooled reactor operating in a process steam/cogeneration mode (HTGR-PS/C) to provide the energy requirements for the SRC-II process. The HTGR-PS/C plant was developed by General Atomic Company (GA) specifically for industries which require energy in the form of both steam and electricity. General Atomic has developed an 1170-MW(t) HTGR-PS/C design which is particularly well suited to industrial applications and is expected to have excellent cost benefits over other sources of energy

The role of Na,K-ATPase/Src-kinase signaling pathway in the vascular wall contaction

DEFF Research Database (Denmark)

Bouzinova, Elena

Aim: Na,K-ATPase is essential for maintaining the transmembrane ion gradient and might initiate various intracellular signaling. These signals possibly act through a modification of the local ion concentrations or via Src-kinase activation. It is known that inhibition of the α-2 isoform of Na......,K-ATPase by ouabain elevates blood pressure. Consequently, ouabain was shown to potentiate arterial contraction in vitro. In contrast, we have demonstrated that siRNA-induced down-regulation of the α-2 isoform Na,K-ATPase expression reduced arterial sensitivity to agonist stimulation and prevented the effect......) phosphorylation assay. Down-regulation of the α-2 isoform Na,K-ATPase prevented the inhibitory effect of Src inhibitors on arterial contraction. Conclusions: The pro-contractile action of ouabain-sensitive Na,K-ATPase inhibition is associated with Src-kinase inhibition suggesting the role of this signaling...

Mechanism of c-Src Synergy with the EGFR In Breast Cancer

National Research Council Canada - National Science Library

Tice, David

1999-01-01

... on tumorigenicity and growth of breast tumor cells. Furthermore, we have discovered a mechanism of c-Src synergy with the EGFR and located specific points at which the pathway can be interdicted...

The Interaction of Src Kinase with beta 3 Integrin Tails : A Potential Therapeutic Target in Thrombosis and Cancer

NARCIS (Netherlands)

Huveneers, Stephan; Danen, Erik H. J.

2010-01-01

Activation of Src family kinases is an important event downstream of integrin adhesion signaling in many cell types. A particularly intriguing connection between an integrin and a Src family kinase was first discovered in platelets, where the selective direct interaction of alpha IIb beta 3

Shear stress induces cell apoptosis via a c-Src-phospholipase D-mTOR signaling pathway in cultured podocytes

Energy Technology Data Exchange (ETDEWEB)

Huang, Chunfa, E-mail: chunfa.huang@case.edu [Louis Stokes Cleveland Veteran Affairs Medical Center, Case Western Reserve University (United States); Department of Medicine, Case Western Reserve University (United States); Rammelkamp Center for Research and Education, MetroHealth System Campus, Cleveland, OH 44106 (United States); Bruggeman, Leslie A. [Department of Medicine, Case Western Reserve University (United States); Rammelkamp Center for Research and Education, MetroHealth System Campus, Cleveland, OH 44106 (United States); Hydo, Lindsey M. [Louis Stokes Cleveland Veteran Affairs Medical Center, Case Western Reserve University (United States); Miller, R. Tyler [Louis Stokes Cleveland Veteran Affairs Medical Center, Case Western Reserve University (United States); Department of Medicine, Case Western Reserve University (United States); Rammelkamp Center for Research and Education, MetroHealth System Campus, Cleveland, OH 44106 (United States)

2012-06-10

The glomerular capillary wall, composed of endothelial cells, the glomerular basement membrane and the podocytes, is continually subjected to hemodynamic force arising from tractional stress due to blood pressure and shear stress due to blood flow. Exposure of glomeruli to abnormal hemodynamic force such as hyperfiltration is associated with glomerular injury and progressive renal disease, and the conversion of mechanical stimuli to chemical signals in the regulation of the process is poorly understood in podocytes. By examining DNA fragmentation, apoptotic nuclear changes and cytochrome c release, we found that shear stress induced cell apoptosis in cultured podocytes. Meanwhile, podocytes exposed to shear stress also stimulated c-Src phosphorylation, phospholipase D (PLD) activation and mammalian target of rapamycin (mTOR) signaling. Using the antibodies against c-Src, PLD{sub 1}, and PLD{sub 2} to perform reciprocal co-immunoprecipitations and in vitro PLD activity assay, our data indicated that c-Src interacted with and activated PLD{sub 1} but not PLD{sub 2}. The inhibition of shear stress-induced c-Src phosphorylation by PP{sub 2} (a specific inhibitor of c-Src kinase) resulted in reduced PLD activity. Phosphatidic acid, produced by shear stress-induced PLD activation, stimulated mTOR signaling, and caused podocyte hypertrophy and apoptosis.

ER-bound protein tyrosine phosphatase PTP1B interacts with Src at the plasma membrane/substrate interface.

Directory of Open Access Journals (Sweden)

Melisa C Monteleone

Full Text Available PTP1B is an endoplasmic reticulum (ER anchored enzyme whose access to substrates is partly dependent on the ER distribution and dynamics. One of these substrates, the protein tyrosine kinase Src, has been found in the cytosol, endosomes, and plasma membrane. Here we analyzed where PTP1B and Src physically interact in intact cells, by bimolecular fluorescence complementation (BiFC in combination with temporal and high resolution microscopy. We also determined the structural basis of this interaction. We found that BiFC signal is displayed as puncta scattered throughout the ER network, a feature that was enhanced when the substrate trapping mutant PTP1B-D181A was used. Time-lapse and co-localization analyses revealed that BiFC puncta did not correspond to vesicular carriers; instead they localized at the tip of dynamic ER tubules. BiFC puncta were retained in ventral membrane preparations after cell unroofing and were also detected within the evanescent field of total internal reflection fluorescent microscopy (TIRFM associated to the ventral membranes of whole cells. Furthermore, BiFC puncta often colocalized with dark spots seen by surface reflection interference contrast (SRIC. Removal of Src myristoylation and polybasic motifs abolished BiFC. In addition, PTP1B active site and negative regulatory tyrosine 529 on Src were primary determinants of BiFC occurrence, although the SH3 binding motif on PTP1B also played a role. Our results suggest that ER-bound PTP1B dynamically interacts with the negative regulatory site at the C-terminus of Src at random puncta in the plasma membrane/substrate interface, likely leading to Src activation and recruitment to adhesion complexes. We postulate that this functional ER/plasma membrane crosstalk could apply to a wide array of protein partners, opening an exciting field of research.

Letrozole regulates actin cytoskeleton polymerization dynamics in a SRC-1 dependent manner in the hippocampus of mice.

Science.gov (United States)

Zhao, Yangang; Yu, Yanlan; Zhang, Yuanyuan; He, Li; Qiu, Linli; Zhao, Jikai; Liu, Mengying; Zhang, Jiqiang

2017-03-01

In the hippocampus, local estrogens (E 2 ) derived from testosterone that is catalyzed by aromatase play important roles in the regulation of hippocampal neural plasticity, but the underlying mechanisms remain unclear. The actin cytoskeleton contributes greatly to hippocampal synaptic plasticity; however, whether it is regulated by local E 2 and the related mechanisms remain to be elucidated. In this study, we first examined the postnatal developmental profiles of hippocampal aromatase and specific proteins responsible for actin cytoskeleton dynamics. Then we used aromatase inhibitor letrozole (LET) to block local E 2 synthesis and examined the changes of these proteins and steroid receptor coactivator-1 (SRC-1), the predominant coactivator for steroid nuclear receptors. Finally, SRC-1 specific RNA interference was used to examine the effects of SRC-1 on the expression of these actin remodeling proteins. The results showed a V-type profile for aromatase and increased profiles for actin cytoskeleton proteins in both male and female hippocampus without obvious sex differences. LET treatment dramatically decreased the F-actin/G-actin ratio, the expression of Rictor, phospho-AKT (ser473), Profilin-1, phospho-Cofilin (Ser3), and SRC-1 in a dose-dependent manner. In vitro studies demonstrated that LET induced downregulation of these proteins could be reversed by E 2 , and E 2 induced increase of these proteins were significantly suppressed by SRC-1 shRNA interference. These results for the first time clearly demonstrated that local E 2 inhibition could induce aberrant actin polymerization; they also showed an important role of SRC-1 in the mediation of local E 2 action on hippocampal synaptic plasticity by regulation of actin cytoskeleton dynamics. Copyright © 2016 Elsevier Ltd. All rights reserved.

77 FR 4581 - Alaska Region's Subsistence Resource Commission (SRC) Program

Science.gov (United States)

2012-01-30

... DEPARTMENT OF THE INTERIOR National Park Service [NPS-AKR-LACL; 9924-PYS] Alaska Region's... public meeting for the National Park Service (NPS) Alaska Region's Subsistence Resource Commission (SRC... Meeting Debora R. Cooper, Associate Regional Director, Resources and Subsistence, Alaska Region. [FR Doc...

Evidence for in vivo phosphorylation of the Grb2 SH2-domain binding site on focal adhesion kinase by Src-family protein-tyrosine kinases.

Science.gov (United States)

Schlaepfer, D D; Hunter, T

1996-10-01

Focal adhesion kinase (FAK) is a nonreceptor protein-tyrosine kinase (PTK) that associates with integrin receptors and participates in extracellular matrix-mediated signal transduction events. We showed previously that the c-Src nonreceptor PTK and the Grb2 SH2/SH3 adaptor protein bound directly to FAK after fibronectin stimulation (D. D. Schlaepfer, S.K. Hanks, T. Hunter, and P. van der Geer, Nature [London] 372:786-791, 1994). Here, we present evidence that c-Src association with FAK is required for Grb2 binding to FAK. Using a tryptic phosphopeptide mapping approach, the in vivo phosphorylation of the Grb2 binding site on FAK (Tyr-925) was detected after fibronectin stimulation of NIH 3T3 cells and was constitutively phosphorylated in v-Src-transformed NIH 3T3 cells. In vitro, c-Src phosphorylated FAK Tyr-925 in a glutathione S-transferase-FAK C-terminal domain fusion protein, whereas FAK did not. Using epitope-tagged FAK constructs, transiently expressed in human 293 cells, we determined the effect of site-directed mutations on c-Src and Grb2 binding to FAK. Mutation of FAK Tyr-925 disrupted Grb2 binding, whereas mutation of the c-Src binding site on FAK (Tyr-397) disrupted both c-Src and Grb2 binding to FAK in vivo. These results support a model whereby Src-family PTKs are recruited to FAK and focal adhesions following integrin-induced autophosphorylation and exposure of FAK Tyr-397. Src-family binding and phosphorylation of FAK at Tyr-925 creates a Grb2 SH2-domain binding site and provides a link to the activation of the Ras signal transduction pathway. In Src-transformed cells, this pathway may be constitutively activated as a result of FAK Tyr-925 phosphorylation in the absence of integrin stimulation.

77 FR 4579 - Alaska Region's Subsistence Resource Commission (SRC) Program

Science.gov (United States)

2012-01-30

... DEPARTMENT OF THE INTERIOR National Park Service [NPS-AKR-DENA; 9924-PYS] Alaska Region's... public meeting for the National Park Service (NPS) Alaska Region's Subsistence Resource Commission (SRC..., Associate Regional Director, Resources and Subsistence, Alaska Region. [FR Doc. 2012-1877 Filed 1-27-12; 8...

Cell Signaling Associated with Na+/K+-ATPase: Activation of Phosphatidylinositide 3-Kinase IA/Akt by Ouabain Is Independent of Src

Science.gov (United States)

2013-01-01

Exposure of intact cells to selective inhibitors of Na+/K+-ATPase such as ouabain activates several growth-related cell signaling pathways. It has been suggested that the initial event of these pathways is the binding of ouabain to a preexisting complex of Src with Na+/K+-ATPase of the plasma membrane. The aim of this work was to evaluate the role of Src in the ouabain-induced activation of phosphatidylinositide 3-kinase 1A (PI3K1A) and its downstream consequences. When fibroblasts devoid of Src (SYF cells) and controls (Src++ cells) were exposed to ouabain, PI3K1A, Akt, and proliferative growth were similarly stimulated in both cell lines. Ouabain-induced activation of Akt was not prevented by the Src inhibitor PP2. In contrast, ERK1/2 were not activated by ouabain in SYF cells but were stimulated in Src++ cells; this was prevented by PP2. In isolated adult mouse cardiac myocytes, where ouabain induces hypertrophic growth, PP2 also did not prevent ouabain-induced activation of Akt and the resulting hypertrophy. Ouabain-induced increases in the levels of co-immunoprecipitation of the α-subunit of Na+/K+-ATPase with the p85 subunit of PI3K1A were noted in SYF cells, Src++ cells, and adult cardiac myocytes. In conjunction with previous findings, the results presented here indicate that (a) if there is a preformed complex of Src and Na+/K+-ATPase, it is irrelevant to ouabain-induced activation of the PI3K1A/Akt pathway through Na+/K+-ATPase and (b) a more likely, but not established, mechanism of linkage of Na+/K+-ATPase to PI3K1A is the ouabain-induced interaction of a proline-rich domain of the α-subunit of Na+/K+-ATPase with the SH3 domain of the p85 subunit of PI3K1A. PMID:24266852

Fractional distillation as a strategy for reducing the genotoxic potential of SRC-II coal liquids: a status report

Energy Technology Data Exchange (ETDEWEB)

Pelroy, R.A.; Wilson, B.W.

1981-09-01

This report presents results of studies on the effects of fractional distillation on the genotoxic potential of Solvent Refined Coal (SRC-II) liquids. SRC-II source materials and distilled liquids were provided by Pittsburg and Midway Coal Mining Co. Fractional distillations were conducted on products from the P-99 process development unit operating under conditions approximating those anticipated at the SRC-II demonstration facility. Distillation cuts were subjected to chemical fractionation, in vitro bioassay and initial chemical analysis. Findings are discussed as they relate to the temperature at which various distillate cuts were produced. This document is the first of two status reports scheduled for 1981 describing these studies.

Computational Prediction of Hot Spot Residues

Science.gov (United States)

Morrow, John Kenneth; Zhang, Shuxing

2013-01-01

Most biological processes involve multiple proteins interacting with each other. It has been recently discovered that certain residues in these protein-protein interactions, which are called hot spots, contribute more significantly to binding affinity than others. Hot spot residues have unique and diverse energetic properties that make them challenging yet important targets in the modulation of protein-protein complexes. Design of therapeutic agents that interact with hot spot residues has proven to be a valid methodology in disrupting unwanted protein-protein interactions. Using biological methods to determine which residues are hot spots can be costly and time consuming. Recent advances in computational approaches to predict hot spots have incorporated a myriad of features, and have shown increasing predictive successes. Here we review the state of knowledge around protein-protein interactions, hot spots, and give an overview of multiple in silico prediction techniques of hot spot residues. PMID:22316154

Endothelial cell-derived matrix promotes the metabolic functional maturation of hepatocyte via integrin-Src signalling.

Science.gov (United States)

Guo, Xinyue; Li, Weihong; Ma, Minghui; Lu, Xin; Zhang, Haiyan

2017-11-01

The extracellular matrix (ECM) microenvironment is involved in the regulation of hepatocyte phenotype and function. Recently, the cell-derived extracellular matrix has been proposed to represent the bioactive and biocompatible materials of the native ECM. Here, we show that the endothelial cell-derived matrix (EC matrix) promotes the metabolic maturation of human adipose stem cell-derived hepatocyte-like cells (hASC-HLCs) through the activation of the transcription factor forkhead box protein A2 (FOXA2) and the nuclear receptors hepatocyte nuclear factor 4 alpha (HNF4α) and pregnane X receptor (PXR). Reducing the fibronectin content in the EC matrix or silencing the expression of α5 integrin in the hASC-HLCs inhibited the effect of the EC matrix on Src phosphorylation and hepatocyte maturation. The inhibition of Src phosphorylation using the inhibitor PP2 or silencing the expression of Src in hASC-HLCs also attenuated the up-regulation of the metabolic function of hASC-HLCs in a nuclear receptor-dependent manner. These data elucidate integrin-Src signalling linking the extrinsic EC matrix signals and metabolic functional maturation of hepatocyte. This study provides a model for studying the interaction between hepatocytes and non-parenchymal cell-derived matrix. © 2017 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

77 FR 4578 - Alaska Region's Subsistence Resource Commission (SRC) Program

Science.gov (United States)

2012-01-30

... DEPARTMENT OF THE INTERIOR National Park Service [NPS-AKR-ANIA; 9924-PYS] Alaska Region's... public meeting for the National Park Service (NPS) Alaska Region's Subsistence Resource Commission (SRC..., Alaska Region. [FR Doc. 2012-1860 Filed 1-27-12; 8:45 am] BILLING CODE 4310-HE-P ...

Evaluation of effects of phenol recovery on biooxidation and tertiary treatment of SRC-I wastewater. Final technical report

Energy Technology Data Exchange (ETDEWEB)

Mitchell, J.W.; Watt, J.C.; Cowan, W.F.; Schuyler, S.E.

1983-09-01

Addition of phenol recovery to the wastewater treatment scheme in the Baseline Design for the SRC-I Demonstration Plant was evaluated as a major post-Baseline effort. Phenol recovery affects many downstream processes, but this study was designed to assess primarily its effects on biooxidation and subsequent tertiary treatment. Two parallel treatment schemes were set up, one to treat dephenolated wastewaters and the other for processed nondephenolated wastewaters, a simulation of the Baseline Design. The study focused on comparisons of five areas: effluent quality; system stability; the need for continuous, high-dose powdered activated carbon (PAC) augmentation to the bioreactor; minimum bioreactor hydraulic residence time (HRT); and tertiary treatment requirements. The results show that phenol recovery improves the quality of the bioreactor effluent in terms of residual organics and color. With phenol recovery, PAC augmentation is not required; without phenol recovery, PAC is needed to produce a comparable effluent. Dephenolization also enhances the stability of biooxidation, and reduces the minimum HRT required. With tertiary treatment, both schemes can meet the effluent concentrations published in the SRC-I Final Envivornmental Impact Statement, as well as the anticipated effluent limits. However, phenol recovery does provide a wider safety margin and could eliminate the need for some of the tertiary treatment steps. Based solely on the technical merits observed in this study, phenol recovery is recommended. The final selection should, however, also consider economic tradeoffs and results of other studies such as toxicology testing of the effluents. 34 references, 30 figures and 26 tables.

Mutations in the catalytic loop HRD motif alter the activity and function of Drosophila Src64.

Directory of Open Access Journals (Sweden)

Taylor C Strong

Full Text Available The catalytic loop HRD motif is found in most protein kinases and these amino acids are predicted to perform functions in catalysis, transition to, and stabilization of the active conformation of the kinase domain. We have identified mutations in a Drosophila src gene, src64, that alter the three HRD amino acids. We have analyzed the mutants for both biochemical activity and biological function during development. Mutation of the aspartate to asparagine eliminates biological function in cytoskeletal processes and severely reduces fertility, supporting the amino acid's critical role in enzymatic activity. The arginine to cysteine mutation has little to no effect on kinase activity or cytoskeletal reorganization, suggesting that the HRD arginine may not be critical for coordinating phosphotyrosine in the active conformation. The histidine to leucine mutant retains some kinase activity and biological function, suggesting that this amino acid may have a biochemical function in the active kinase that is independent of its side chain hydrogen bonding interactions in the active site. We also describe the phenotypic effects of other mutations in the SH2 and tyrosine kinase domains of src64, and we compare them to the phenotypic effects of the src64 null allele.

Mechanism of c-Src Synergy with the EGFR in Breast Cancer

National Research Council Canada - National Science Library

1998-01-01

.... Specifically, we have shown that kinase-inactive c-Src is able to inhibit tumorigenicity of the IOT 1/2 mouse fibroblast model cells by not phosphorylating the receptor on Tyr 845 in the activation loop of the kinase...

Pro-contractile action of the Na,K-ATPase/Src-kinase signaling pathway in the vascular wall

DEFF Research Database (Denmark)

Bouzinova, Elena; Aalkjær, Christian; Matchkov, Vladimir

Aim: Na,K-ATPase is essential for maintaining the transmembrane ion gradient and might initiate various intracellular signaling. These signals possibly act through a modification of the local ion concentrations or via Src-kinase activation. It is known that inhibition of the α-2 isoform of Na......,K-ATPase by ouabain elevates blood pressure. Consequently, ouabain was shown to potentiate arterial contraction in vitro. In contrast, we have demonstrated that siRNA-induced down-regulation of the α-2 isoform Na,K-ATPase expression reduced arterial sensitivity to agonist stimulation and prevented the effect......) phosphorylation assay. Down-regulation of the α-2 isoform Na,K-ATPase prevented the inhibitory effect of Src inhibitors on arterial contraction. Conclusions: The pro-contractile action of ouabain-sensitive Na,K-ATPase inhibition is associated with Src-kinase inhibition suggesting the role of this signaling...

Drive and the Action Calculation of GetLLM, in Beta-Beat.src

CERN Document Server

Sherman, Alexander Charles

2013-01-01

The Beta-Beat.src program is used to analyze data collected by Beam Position Monitors (BPMs) in accelerators at CERN. The Beams department at CERN uses it to study the behaviour of a beam as it traverses an accelerator, and in particular the LHC. Two pieces of code in Beta-Beat.src are “drive”, a C/C++ program, and “GetLLM”, a python program. This report described the modification of the drive code to be compatible with windows and take advantage of elements of C++, as well as a change in the calculation of the action in GetLLM to reduce its relative uncertainty. The dynamic aperture is recalculated with the new action and sees a reduction in its uncertainty.

A novel disulfide bond in the SH2 Domain of the C-terminal Src kinase controls catalytic activity.

Science.gov (United States)

Mills, Jamie E; Whitford, Paul C; Shaffer, Jennifer; Onuchic, Jose N; Adams, Joseph A; Jennings, Patricia A

2007-02-02

The SH2 domain of the C-terminal Src kinase [Csk] contains a unique disulfide bond that is not present in other known SH2 domains. To investigate whether this unusual disulfide bond serves a novel function, the effects of disulfide bond formation on catalytic activity of the full-length protein and on the structure of the SH2 domain were investigated. The kinase activity of full-length Csk decreases by an order of magnitude upon formation of the disulfide bond in the distal SH2 domain. NMR spectra of the fully oxidized and fully reduced SH2 domains exhibit similar chemical shift patterns and are indicative of similar, well-defined tertiary structures. The solvent-accessible disulfide bond in the isolated SH2 domain is highly stable and far from the small lobe of the kinase domain. However, reduction of this bond results in chemical shift changes of resonances that map to a cluster of residues that extend from the disulfide bond across the molecule to a surface that is in direct contact with the small lobe of the kinase domain in the intact molecule. Normal mode analyses and molecular dynamics calculations suggest that disulfide bond formation has large effects on residues within the kinase domain, most notably within the active-site cleft. Overall, the data indicate that reversible cross-linking of two cysteine residues in the SH2 domain greatly impacts catalytic function and interdomain communication in Csk.

Role of miR-222-3p in c-Src-Mediated Regulation of Osteoclastogenesis

Directory of Open Access Journals (Sweden)

Shinya Takigawa

2016-02-01

Full Text Available MicroRNAs (miRNAs are small non-coding RNAs that play a mostly post-transcriptional regulatory role in gene expression. Using RAW264.7 pre-osteoclast cells and genome-wide expression analysis, we identified a set of miRNAs that are involved in osteoclastogenesis. Based on in silico analysis, we specifically focused on miR-222-3p and evaluated its role in osteoclastogenesis. The results show that the inhibitor of miR-222-3p upregulated the mRNA levels of nuclear factor of activated T-cells, cytoplasmic 1 (NFATc1 and tartrate-resistant acid phosphatase (TRAP, while its mimicking agent downregulated their mRNA levels. Western blot analysis showed that its inhibitor increased the protein levels of TRAP and cathepsin K, while its mimicking agent decreased their levels. Genome-wide mRNA expression analysis in the presence and absence of receptor activator of nuclear factor κ-B ligand (RANKL predicted c-Src as a potential regulatory target of miR-222-3p. Live cell imaging using a fluorescence resonance energy transfer (FRET technique revealed that miR-222-3p acted as an inhibitor of c-Src activity, and a partial silencing of c-Src suppressed RANKL-induced expression of TRAP and cathepsin K, as well as the number of multi-nucleated osteoclasts and their pit formation. Collectively, the study herein demonstrates that miR-222-3p serves as an inhibitor of osteoclastogenesis and c-Src mediates its inhibition of cathepsin K and TRAP.

The deleted in brachydactyly B domain of ROR2 is required for receptor activation by recruitment of Src.

Directory of Open Access Journals (Sweden)

Shiva Akbarzadeh

2008-03-01

Full Text Available The transmembrane receptor 'ROR2' resembles members of the receptor tyrosine kinase family of signalling receptors in sequence but its' signal transduction mechanisms remain enigmatic. This problem has particular importance because mutations in ROR2 are associated with two human skeletal dysmorphology syndromes, recessive Robinow Syndrome (RS and dominant acting Brachydactyly type B (BDB. Here we show, using a constitutive dimerisation approach, that ROR2 exhibits dimerisation-induced tyrosine kinase activity and the ROR2 C-terminal domain, which is deleted in BDB, is required for recruitment and activation of the non-receptor tyrosine kinase Src. Native ROR2 phosphorylation is induced by the ligand Wnt5a and is blocked by pharmacological inhibition of Src kinase activity. Eight sites of Src-mediated ROR2 phosphorylation have been identified by mass spectrometry. Activation via tyrosine phosphorylation of ROR2 receptor leads to its internalisation into Rab5 positive endosomes. These findings show that BDB mutant receptors are defective in kinase activation as a result of failure to recruit Src.

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...

Inhibition of Src by microRNA-23b increases the cisplatin sensitivity of chondrosarcoma cells.

Science.gov (United States)

Huang, Kai; Chen, Jun; Yang, Mo-Song; Tang, Yu-Jun; Pan, Feng

2017-01-01

Chondrosarcomas are malignant cartilage-forming tumors from low-grade to high-grade aggressive tumors characterized by metastasis. Cisplatin is an effective DNA-damaging anti-tumor agent for the treatment against a wide variety of solid tumors. However, chondrosarcomas are notorious for their resistance to conventional chemo- and radio- therapies. In this study, we report miR-23b acts as a tumor suppressor in chondrosarcoma. The expressions of miR-23b are down-regulated in chondrosarcoma patient samples and cell lines compared with adjacent normal tissues and human primary chondrocytes. In addition, overexpression of miR-23b suppresses chondrosarcoma cell proliferation. By comparison of the cisplatin resistant chondrosarcoma cells and parental cells, we observed miR-23b was significantly down regulated in cisplatin resistant cells. Moreover, we demonstrate here Src kinase is a direct target of miR-23b in chondrosarcoma cells. Overexpression of miR-23b suppresses Src-Akt pathway, leading to the sensitization of cisplatin resistant chondrosarcoma cells to cisplatin. This chemo-sensitivity effect by the miR-23b-mediated inhibition of Src-Akt pathway is verified with the restoration of Src kinase in miR-23b-overespressing chondrosarcoma cells, resulting in the acquirement of resistance to cisplatin. In summary, our study reveals a novel role of miR-23b in cisplatin resistance in chondrosarcoma and will contribute to the development of the microRNA-targeted anti-cancer therapeutics.

Empowering Provenance in Data Integration

Science.gov (United States)

Kondylakis, Haridimos; Doerr, Martin; Plexousakis, Dimitris

The provenance of data has recently been recognized as central to the trust one places in data. This paper presents a novel framework in order to empower provenance in a mediator based data integration system. We use a simple mapping language for mapping schema constructs, between an ontology and relational sources, capable to carry provenance information. This language extends the traditional data exchange setting by translating our mapping specifications into source-to-target tuple generating dependencies (s-t tgds). Then we define formally the provenance information we want to retrieve i.e. annotation, source and tuple provenance. We provide three algorithms to retrieve provenance information using information stored on the mappings and the sources. We show the feasibility of our solution and the advantages of our framework.

Potential benefits of commercial willow Short Rotation Coppice (SRC) for farm-scale plant and invertebrate communities in the agri-environment

Energy Technology Data Exchange (ETDEWEB)

Rowe, Rebecca L.; Hanley, Mick E.; Goulson, Dave; Clarke, Donna J.; Doncaster, C. Patrick; Taylor, Gail [University of Southampton, Faculty of Natural and Environmental Sciences, Life Sciences Building, Southampton, S017 1BJ (United Kingdom)

2011-01-15

The cultivation of bioenergy crops (BECs) represents a significant land-use change in agri-environments, but their deployment has raised important issues globally regarding possible impacts on biodiversity. Few studies however, have systematically examined the effect of commercial scale bioenergy plantations on biodiversity in agri-ecosystems. In this study we investigate how the abundance and diversity of two key components of farmland biodiversity (ground flora and winged invertebrates) varied between mature willow Short Rotation Coppice (SRC) and two alternative land-use options (arable crops and set-aside land). Although the abundance of winged invertebrates was similar across all land-uses, taxonomic composition varied markedly. Hymenoptera and large Hemiptera (>5 mm) were more abundant in willow SRC than in arable or set-aside. Similarly although plant species richness was greater in set-aside, our data show that willow SRC supports a different plant community to the other land-uses, being dominated by competitive perennial species such as Elytrigia repens and Urtica dioica. Our results suggest that under current management practices a mixed farming system incorporating willow SRC can benefit native farm-scale biodiversity. In particular the reduced disturbance in willow SRC allows the persistence of perennial plant species, potentially providing a stable refuge and food sources for invertebrates. In addition, increased Hymenoptera abundance in willow SRC could potentially have concomitant effects on ecosystem processes, as many members of this Order are important pollinators of crop plants or otherwise fulfil an important beneficial role as predators or parasites of crop pests. (author)

Vascular endothelial growth factor receptor-1 mediates migration of human colorectal carcinoma cells by activation of Src family kinases

Science.gov (United States)

Lesslie, D P; Summy, J M; Parikh, N U; Fan, F; Trevino, J G; Sawyer, T K; Metcalf, C A; Shakespeare, W C; Hicklin, D J; Ellis, L M; Gallick, G E

2006-01-01

Vascular endothelial growth factor (VEGF) is the predominant pro-angiogenic cytokine in human malignancy, and its expression correlates with disease recurrence and poor outcomes in patients with colorectal cancer. Recently, expression of vascular endothelial growth factor receptors (VEGFRs) has been observed on tumours of epithelial origin, including those arising in the colon, but the molecular mechanisms governing potential VEGF-driven biologic functioning in these tumours are not well characterised. In this report, we investigated the role of Src family kinases (SFKs) in VEGF-mediated signalling in human colorectal carcinoma (CRC) cell lines. Vascular endothelial growth factor specifically activated SFKs in HT29 and KM12L4 CRC cell lines. Further, VEGF stimulation resulted in enhanced cellular migration, which was effectively blocked by pharmacologic inhibition of VEGFR-1 or Src kinase. Correspondingly, migration studies using siRNA clones with reduced Src expression confirmed the requirement for Src in VEGF-induced migration in these cells. Furthermore, VEGF treatment enhanced VEGFR-1/SFK complex formation and increased tyrosine phosphorylation of focal adhesion kinase, p130 cas and paxillin. Finally, we demonstrate that VEGF-induced migration is not due, at least in part, to VEGF acting as a mitogen. These results suggest that VEGFR-1 promotes migration of tumour cells through a Src-dependent pathway linked to activation of focal adhesion components that regulate this process. PMID:16685275

Solvent refined coal (SRC) process: trace elements research and development report no. 53, interim report no. 34. Volume III. Pilot plant development work part 6: the fate of trace elements in the SRC process for the period, August 1, 1977 - February 28, 1979

International Nuclear Information System (INIS)

Filby, R.H.; Khalil, S.R.; Grimm, C.A.; Ekabaram, V.; Hunt, M.L.

1980-12-01

This work reports the results of neutron activation analysis determination of the fate of trace elements in the SRC II process. Six coals were studied for their behavior in material balance runs carried out at the Fort Lewis Pilot Plant. The distribution of trace elements among products and input streams was determined by thermal neutron activation analysis using thermal neutron flux of 8 x 10 12 neutrons cm -2 sec -1 followed by Ge(Li) gamma ray spectroscopy. National Bureau of Standards Standard Reference Materials (SRM) were used to evaluate the analytical precision and accuracy of the methods used. For each material balance study the trace element input stream was taken as ground coal and the output streams were vacuum bottoms, SRC II product oil, and process water. In addition to these major components, oils, sludges and waters from liquid-liquid separators, effluent waters, biosludges, and by-product sulfur were also analyzed. Concerning the distribution of trace elements in the SRC II process, it was found that the vacuum bottoms was the major sink for all trace element studied, with the exception of Hg. Much lower trace element concentrations (except for Hg) were found in the SRC II product oil relative to the vacuum bottoms or the feed coal, irrespective of coal type. The results indicate excellent balances for the elements studied, except for Hg. Except for Hg, Se, and C1, the SRC II product and process waters contributed less than 1% of the elemental balances for light oil fractions and process waters indicates that Hg, and to a lesser degree As, Se, and Sb, exhibited volatile behavior in the SRC II process but that the degree of volatility is strongly dependent on conditions or coal type

77 FR 4580 - Alaska Region's Subsistence Resource Commission (SRC) Program

Science.gov (United States)

2012-01-30

..., February 14, 2012. The meeting will start at 9 a.m. and conclude at 5 p.m. or until business is completed.... Old Business a. Subsistence Collections and Uses of Shed or Discarded Animal & Plants Environmental.... New Business 12. Public and other Agency Comments 13. SRC Work Session 14. Select Time and Location...

Logical provenance in data-oriented workflows?

KAUST Repository

Ikeda, R.

2013-04-01

We consider the problem of defining, generating, and tracing provenance in data-oriented workflows, in which input data sets are processed by a graph of transformations to produce output results. We first give a new general definition of provenance for general transformations, introducing the notions of correctness, precision, and minimality. We then determine when properties such as correctness and minimality carry over from the individual transformations\\' provenance to the workflow provenance. We describe a simple logical-provenance specification language consisting of attribute mappings and filters. We provide an algorithm for provenance tracing in workflows where logical provenance for each transformation is specified using our language. We consider logical provenance in the relational setting, observing that for a class of Select-Project-Join (SPJ) transformations, logical provenance specifications encode minimal provenance. We have built a prototype system supporting the features and algorithms presented in the paper, and we report a few preliminary experimental results. © 2013 IEEE.

Src Family Kinases and p38 Mitogen-Activated Protein Kinases Regulate Pluripotent Cell Differentiation in Culture.

Directory of Open Access Journals (Sweden)

Boon Siang Nicholas Tan

Full Text Available Multiple pluripotent cell populations, which together comprise the pluripotent cell lineage, have been identified. The mechanisms that control the progression between these populations are still poorly understood. The formation of early primitive ectoderm-like (EPL cells from mouse embryonic stem (mES cells provides a model to understand how one such transition is regulated. EPL cells form from mES cells in response to l-proline uptake through the transporter Slc38a2. Using inhibitors of cell signaling we have shown that Src family kinases, p38 MAPK, ERK1/2 and GSK3β are required for the transition between mES and EPL cells. ERK1/2, c-Src and GSK3β are likely to be enforcing a receptive, primed state in mES cells, while Src family kinases and p38 MAPK are involved in the establishment of EPL cells. Inhibition of these pathways prevented the acquisition of most, but not all, features of EPL cells, suggesting that other pathways are required. L-proline activation of differentiation is mediated through metabolism and changes to intracellular metabolite levels, specifically reactive oxygen species. The implication of multiple signaling pathways in the process suggests a model in which the context of Src family kinase activation determines the outcomes of pluripotent cell differentiation.

Src Family Kinases and p38 Mitogen-Activated Protein Kinases Regulate Pluripotent Cell Differentiation in Culture

Science.gov (United States)

Tan, Boon Siang Nicholas; Kwek, Joly; Wong, Chong Kum Edwin; Saner, Nicholas J.; Yap, Charlotte; Felquer, Fernando; Morris, Michael B.; Gardner, David K.; Rathjen, Peter D.; Rathjen, Joy

2016-01-01

Multiple pluripotent cell populations, which together comprise the pluripotent cell lineage, have been identified. The mechanisms that control the progression between these populations are still poorly understood. The formation of early primitive ectoderm-like (EPL) cells from mouse embryonic stem (mES) cells provides a model to understand how one such transition is regulated. EPL cells form from mES cells in response to l-proline uptake through the transporter Slc38a2. Using inhibitors of cell signaling we have shown that Src family kinases, p38 MAPK, ERK1/2 and GSK3β are required for the transition between mES and EPL cells. ERK1/2, c-Src and GSK3β are likely to be enforcing a receptive, primed state in mES cells, while Src family kinases and p38 MAPK are involved in the establishment of EPL cells. Inhibition of these pathways prevented the acquisition of most, but not all, features of EPL cells, suggesting that other pathways are required. L-proline activation of differentiation is mediated through metabolism and changes to intracellular metabolite levels, specifically reactive oxygen species. The implication of multiple signaling pathways in the process suggests a model in which the context of Src family kinase activation determines the outcomes of pluripotent cell differentiation. PMID:27723793

Special Section: The third provenance challenge on using the open provenance model for interoperability

NARCIS (Netherlands)

Simmhan, Y; Groth, P.T.; Moreau, L

2011-01-01

The third provenance challenge was organized to evaluate the efficacy of the Open Provenance Model (OPM) in representing and sharing provenance with the goal of improving the specification. A data loading scientific workflow that ingests data files into a relational database for the Pan-STARRS sky

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.

Tyrosine Kinase Inhibition in HPV-related Squamous Cell Carcinoma Reveals Beneficial Expression of cKIT and Src.

Science.gov (United States)

Kramer, Benedikt; Kneissle, Marcel; Birk, Richard; Rotter, Nicole; Aderhold, Christoph

2018-05-01

Therapeutic options of locally advanced or metastatic head and neck squamous cell carcinoma (HNSCC) are limited. Src and cKIT are key protein regulators for local tumor progression. The aim of the study was to investigate the therapeutic potential of targeted therapies in human squamous cell carcinoma (HNSCC) in vitro. Therefore, the influence of the selective tyrosine kinase inhibitors niotinib, dasatinib, erlotinib, gefitinib and afatinib on Src and cKIT expression in Human papilloma virus (HPV)-positive and HPV-negative squamous cancer cells (SCC) was analyzed in vitro. ELISA was performed to evaluate the expression of Src and cKIT under the influence of nilotinib, dasatinib, erlotinib, gefitinib and afatinib (10 μmol/l) in HPV-negative and HPV-positive SCC (24-96 h of incubation). Gefitinib significantly increased cKIT expression in HPV-positive and HPV-negative cells whereas nilotinib and afatinib decreased cKIT expression in HPV-positive SCC. The influence of tyrosine kinase inhibitors in HPV-negative SCC was marginal. Surprisingly, Src expression was significantly increased by all tested tyrosine kinase inhibitors in HPV-positive SCC. The results revealed beneficial and unexpected information concerning the interaction of selective tyrosine kinase inhibitors and the tumor biology of HNSCC. Copyright© 2018, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.

Computational prediction of protein hot spot residues.

Science.gov (United States)

Morrow, John Kenneth; Zhang, Shuxing

2012-01-01

Most biological processes involve multiple proteins interacting with each other. It has been recently discovered that certain residues in these protein-protein interactions, which are called hot spots, contribute more significantly to binding affinity than others. Hot spot residues have unique and diverse energetic properties that make them challenging yet important targets in the modulation of protein-protein complexes. Design of therapeutic agents that interact with hot spot residues has proven to be a valid methodology in disrupting unwanted protein-protein interactions. Using biological methods to determine which residues are hot spots can be costly and time consuming. Recent advances in computational approaches to predict hot spots have incorporated a myriad of features, and have shown increasing predictive successes. Here we review the state of knowledge around protein-protein interactions, hot spots, and give an overview of multiple in silico prediction techniques of hot spot residues.

The spatiotemporal pattern of Src activation at lipid rafts revealed by diffusion-corrected FRET imaging.

Directory of Open Access Journals (Sweden)

Shaoying Lu

2008-07-01

Full Text Available Genetically encoded biosensors based on fluorescence resonance energy transfer (FRET have been widely applied to visualize the molecular activity in live cells with high spatiotemporal resolution. However, the rapid diffusion of biosensor proteins hinders a precise reconstruction of the actual molecular activation map. Based on fluorescence recovery after photobleaching (FRAP experiments, we have developed a finite element (FE method to analyze, simulate, and subtract the diffusion effect of mobile biosensors. This method has been applied to analyze the mobility of Src FRET biosensors engineered to reside at different subcompartments in live cells. The results indicate that the Src biosensor located in the cytoplasm moves 4-8 folds faster (0.93+/-0.06 microm(2/sec than those anchored on different compartments in plasma membrane (at lipid raft: 0.11+/-0.01 microm(2/sec and outside: 0.18+/-0.02 microm(2/sec. The mobility of biosensor at lipid rafts is slower than that outside of lipid rafts and is dominated by two-dimensional diffusion. When this diffusion effect was subtracted from the FRET ratio images, high Src activity at lipid rafts was observed at clustered regions proximal to the cell periphery, which remained relatively stationary upon epidermal growth factor (EGF stimulation. This result suggests that EGF induced a Src activation at lipid rafts with well-coordinated spatiotemporal patterns. Our FE-based method also provides an integrated platform of image analysis for studying molecular mobility and reconstructing the spatiotemporal activation maps of signaling molecules in live cells.

The role of c-Src in the invasion and metastasis of hepatocellular carcinoma cells induced by association of cell surface GRP78 with activated α2M

International Nuclear Information System (INIS)

Zhao, Song; Li, Hongdan; Wang, Qingjun; Su, Chang; Wang, Guan; Song, Huijuan; Zhao, Liang; Luan, Zhidong; Su, Rongjian

2015-01-01

Emerging data have suggested that cell surface GRP78 is a multifunctional receptor and has been linked to proliferative and antiapoptotic signaling cascades. Activated α 2− macroglobin (α 2 M*) is a natural circulating ligand of cell surface GRP78. Association of cell surface GRP78 with α 2 M* is involved in the regulation of cell proliferation, survival and apoptosis in human cancers. The invasion and metastasis of HCC cells were examined using transwell and wound healing assay; Cell surface expression of GRP78 was detected by in cell western assay. Translocation of GRP78 from cytosol to cell surface was observed by transfection of GRP78-EGFP plus TRIRC-WGA staining. The levels of Src, phosphor-Src, FAK, phospho-FAK, EGFR, phospho-EGFR, phospho-Cortactin, phospho-Paxillin were determined by western blot. Cell surface expression of GRP78 in HCC tissue samples was observed by immunofluorescence. The distribution of Paxillin and Cortactin in HCC cells was also observed by immunofluorescence. The interaction between GRP78 and Src were detected by far-western blot, co-immunoprecipitation and GST pulldown. GRP78 mRNA was detected by RT-PCR. In the current study, we showed that association of cell surface GRP78 with α 2 M* stimulated the invasion and metastasis of HCC. Cell surface GRP78 could interact directly with c-Src, promoted the phosphorylation of c-Src at Y416. Inhibition of the tyrosine kinase activity of c-Src with PP2 reverted the stimulatory effect caused by association of cell surface GRP78 with α 2 M*. Moreover, association of cell surface GRP78 with α 2 M* facilitates the interaction between EGFR and c-Src and consequently phosphorylated EGFR at Y1101 and Y845, promoting the invasion and metastasis of HCCs. However, inhibition of the tyrosine kinase of c-Src do not affect the interaction between EGFR and Src. c-Src plays a critical role in the invasion and metastasis of HCC induced by association of cell surface GRP78 with α 2 M*. Cell surface GRP

Dendritic cells sensitize TCRs through self-MHC-mediated Src family kinase activation

Czech Academy of Sciences Publication Activity Database

Meraner, P.; Hořejší, Václav; Wolpl, A.; Fischer, G.F.; Stingl, G.; Maurer, D.

2007-01-01

Roč. 178, č. 4 (2007), s. 2262-2271 ISSN 0022-1767 Institutional research plan: CEZ:AV0Z50520514 Keywords : TCR * dendritic cells * Src kinases Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 6.068, year: 2007

A dynamically coupled allosteric network underlies binding cooperativity in Src kinase.

Science.gov (United States)

Foda, Zachariah H; Shan, Yibing; Kim, Eric T; Shaw, David E; Seeliger, Markus A

2015-01-20

Protein tyrosine kinases are attractive drug targets because many human diseases are associated with the deregulation of kinase activity. However, how the catalytic kinase domain integrates different signals and switches from an active to an inactive conformation remains incompletely understood. Here we identify an allosteric network of dynamically coupled amino acids in Src kinase that connects regulatory sites to the ATP- and substrate-binding sites. Surprisingly, reactants (ATP and peptide substrates) bind with negative cooperativity to Src kinase while products (ADP and phosphopeptide) bind with positive cooperativity. We confirm the molecular details of the signal relay through the allosteric network by biochemical studies. Experiments on two additional protein tyrosine kinases indicate that the allosteric network may be largely conserved among these enzymes. Our work provides new insights into the regulation of protein tyrosine kinases and establishes a potential conduit by which resistance mutations to ATP-competitive kinase inhibitors can affect their activity.

Solvent-refined-coal (SRC) process. Volume II. Sections V-XIV. Final report

Energy Technology Data Exchange (ETDEWEB)

1982-05-01

This report documents the completion of development work on the Solvent Refined Coal Process by The Pittsburgh and Midway Coal Mining Co. The work was initiated in 1966 under Office of Coal Research, US Department of Interior, Contract No. 14-01-0001-496 and completed under US Department of Energy Contract No. DE-AC05-79ET10104. This report discusses work leading to the development of the SRC-I and SRC-II processes, construction of the Fort Lewis Pilot Plant for the successful development of these processes, and results from the operation of this pilot plant. Process design data generated on a 1 ton-per-day Process Development Unit, bench-scale units and through numerous research projects in support of the design of major demonstration plants are also discussed in summary form and fully referenced in this report.

Inhibition of Src kinase activity attenuates amyloid associated microgliosis in a murine model of Alzheimer’s disease

Directory of Open Access Journals (Sweden)

Dhawan Gunjan

2012-07-01

Full Text Available Abstract Background Microglial activation is an important histologic characteristic of the pathology of Alzheimer’s disease (AD. One hypothesis is that amyloid beta (Aβ peptide serves as a specific stimulus for tyrosine kinase-based microglial activation leading to pro-inflammatory changes that contribute to disease. Therefore, inhibiting Aβ stimulation of microglia may prove to be an important therapeutic strategy for AD. Methods Primary murine microglia cultures and the murine microglia cell line, BV2, were used for stimulation with fibrillar Aβ1-42. The non-receptor tyrosine kinase inhibitor, dasatinib, was used to treat the cells to determine whether Src family kinase activity was required for the Aβ stimulated signaling response and subsequent increase in TNFα secretion using Western blot analysis and enzyme-linked immunosorbent assay (ELISA, respectively. A histologic longitudinal analysis was performed using an AD transgenic mouse model, APP/PS1, to determine an age at which microglial protein tyrosine kinase levels increased in order to administer dasatinib via mini osmotic pump diffusion. Effects of dasatinib administration on microglial and astroglial activation, protein phosphotyrosine levels, active Src kinase levels, Aβ plaque deposition, and spatial working memory were assessed via immunohistochemistry, Western blot, and T maze analysis. Results Aβ fibrils stimulated primary murine microglia via a tyrosine kinase pathway involving Src kinase that was attenuated by dasatinib. Dasatinib administration to APP/PS1 mice decreased protein phosphotyrosine, active Src, reactive microglia, and TNFα levels in the hippocampus and temporal cortex. The drug had no effect on GFAP levels, Aβ plaque load, or the related tyrosine kinase, Lyn. These anti-inflammatory changes correlated with improved performance on the T maze test in dasatinib infused animals compared to control animals. Conclusions These data suggest that amyloid

Global phosphotyrosine proteomics identifies PKCδ as a marker of responsiveness to Src inhibition in colorectal cancer.

Directory of Open Access Journals (Sweden)

Eliot T McKinley

Full Text Available Sensitive and specific biomarkers of protein kinase inhibition can be leveraged to accelerate drug development studies in oncology by associating early molecular responses with target inhibition. In this study, we utilized unbiased shotgun phosphotyrosine (pY proteomics to discover novel biomarkers of response to dasatinib, a small molecule Src-selective inhibitor, in preclinical models of colorectal cancer (CRC. We performed unbiased mass spectrometry shotgun pY proteomics to reveal the pY proteome of cultured HCT-116 colonic carcinoma cells, and then extended this analysis to HCT-116 xenograft tumors to identify pY biomarkers of dasatinib-responsiveness in vivo. Major dasatinib-responsive pY sites in xenograft tumors included sites on delta-type protein kinase C (PKCδ, CUB-domain-containing protein 1 (CDCP1, Type-II SH2-domain-containing inositol 5-phosphatase (SHIP2, and receptor protein-tyrosine phosphatase alpha (RPTPα. The pY313 site PKCδ was further supported as a relevant biomarker of dasatinib-mediated Src inhibition in HCT-116 xenografts by immunohistochemistry and immunoblotting with a phosphospecific antibody. Reduction of PKCδ pY313 was further correlated with dasatinib-mediated inhibition of Src and diminished growth as spheroids of a panel of human CRC cell lines. These studies reveal PKCδ pY313 as a promising readout of Src inhibition in CRC and potentially other solid tumors and may reflect responsiveness to dasatinib in a subset of colorectal cancers.

File Level Provenance Tracking in CMS

CERN Document Server

Jones, C D; Paterno, M; Sexton-Kennedy, L; Tanenbaum, W; Riley, D S

2009-01-01

The CMS off-line framework stores provenance information within CMS's standard ROOT event data files. The provenance information is used to track how each data product was constructed, including what other data products were read to do the construction. We will present how the framework gathers the provenance information, the efforts necessary to minimise the space used to store the provenance in the file and the tools that will be available to use the provenance.

Physical and functional interactions between SH2 and SH3 domains of the Src family protein tyrosine kinase p59fyn

NARCIS (Netherlands)

Panchamoorthy, G.; Fukazawa, T.; Stolz, L.; Payne, G.; Reedquist, K.; Shoelson, S.; Songyang, Z.; Cantley, L.; Walsh, C.; Band, H.

1994-01-01

The Src family protein tyrosine kinases participate in signalling through cell surface receptors that lack intrinsic tyrosine kinase domains. All nine members of this family possess adjacent Src homology (SH2 and SH3) domains, both of which are essential for repression of the enzymatic activity. The

Provenance management in Swift with implementation details.

Energy Technology Data Exchange (ETDEWEB)

Gadelha, L. M. R; Clifford, B.; Mattoso, M.; Wilde, M.; Foster, I. (Mathematics and Computer Science); ( CLS-CI); (Federal Univ. of Rio de Janeiro); (National Lab. for Scientific Computing, Brazil); (Univ. of Chicago)

2011-04-01

The Swift parallel scripting language allows for the specification, execution and analysis of large-scale computations in parallel and distributed environments. It incorporates a data model for recording and querying provenance information. In this article we describe these capabilities and evaluate interoperability with other systems through the use of the Open Provenance Model. We describe Swift's provenance data model and compare it to the Open Provenance Model. We also describe and evaluate activities performed within the Third Provenance Challenge, which consisted of implementing a specific scientific workflow, capturing and recording provenance information of its execution, performing provenance queries, and exchanging provenance information with other systems. Finally, we propose improvements to both the Open Provenance Model and Swift's provenance system.

Proven Weight Loss Methods

Science.gov (United States)

Fact Sheet Proven Weight Loss Methods What can weight loss do for you? Losing weight can improve your health in a number of ways. It can lower ... at www.hormone.org/Spanish . Proven Weight Loss Methods Fact Sheet www.hormone.org

CMOS-based active RC sinusoidal oscillator with four-phase quadrature outputs and single-resistance-controlled (SRC) tuning laws

OpenAIRE

Lahiri, Abhirup; Herencsár, Norbert

2012-01-01

This paper proposes a very compact CMOS realization of active RC sinusoidal oscillator capable of generating four quadrature voltage outputs. The oscillator is based on the cascade of lossless and lossy integrators in loop. The governing laws for the condition of oscillation (CO) and the frequency of oscillation (FO) are single-resistance-controlled (SRC) and which allow independent FO tuning. Unlike previously reported SRC-based sinusoidal oscillators based on the active building block (ABB)...

Active Provenance in Data-intensive Research

Science.gov (United States)

Spinuso, Alessandro; Mihajlovski, Andrej; Filgueira, Rosa; Atkinson, Malcolm

2017-04-01

Scientific communities are building platforms where the usage of data-intensive workflows is crucial to conduct their research campaigns. However managing and effectively support the understanding of the 'live' processes, fostering computational steering, sharing and re-use of data and methods, present several bottlenecks. These are often caused by the poor level of documentation on the methods and the data and how users interact with it. This work wants to explore how in such systems, flexibility in the management of the provenance and its adaptation to the different users and application contexts can lead to new opportunities for its exploitation, improving productivity. In particular, this work illustrates a conceptual and technical framework enabling tunable and actionable provenance in data-intensive workflow systems in support of reproducible science. It introduces the concept of Agile data-intensive systems to define the characteristic of our target platform. It shows a novel approach to the integration of provenance mechanisms, offering flexibility in the scale and in the precision of the provenance data collected, ensuring its relevance to the domain of the data-intensive task, fostering its rapid exploitation. The contributions address aspects of the scale of the provenance records, their usability and active role in the research life-cycle. We will discuss the use of dynamically generated provenance types as the approach for the integration of provenance mechanisms into a data-intensive workflow system. Enabling provenance can be transparent to the workflow user and developer, as well as fully controllable and customisable, depending from their expertise and the application's reproducibility, monitoring and validation requirements. The API that allows the realisation and adoption of a provenance type is presented, especially for what concerns the support of provenance profiling, contextualisation and precision. An actionable approach to provenance

Sustained expression of steroid receptor coactivator SRC-2/TIF-2 is associated with better prognosis in malignant pleural mesothelioma.

LENUS (Irish Health Repository)

Jennings, Cormac J

2012-02-01

INTRODUCTION: Estrogen receptor beta (ERbeta) overexpression by malignant pleural mesothelioma (MPM) tumor cells correlates with enhanced patient survival. ER-regulated transcription is mediated by the p160 family of steroid receptor coactivators (SRCs), and SRC isoform overexpression is associated with worse prognosis in many steroid-related malignancies. The aim of this study was to establish whether SRC isoform expression varied between individual MPM tumors with positive or negative prognostic significance. METHODS: Immunohistochemical analysis of tumor biopsies from 89 subjects with confirmed histological diagnosis of MPM and biopsies from 3 normal control subjects was performed to detect the expression of SRC-1, SRC-2 (TIF-2), SRC-3 (AIB-1), and ERbeta. Allred scores for expression of ERbeta and each of the SRCs were determined, and Kaplan-Meier survival curves were calculated to correlate biomarker expression, gender, and histology type with postdiagnosis survival. RESULTS: ERbeta and all the SRCs were expressed at high levels in normal pleural mesothelium, and expression of each biomarker was reduced or lost in a subset of the MPM subjects; however, postdiagnosis survival only significantly correlated with TIF-2 expression. Low or intermediate expression of TIF-2 correlated with reduced median postdiagnosis survival (9 months) compared with those subjects whose tumors highly expressed TIF-2 (20 months) (p = 0.036, log-rank test). CONCLUSIONS: Maintained high expression of TIF-2 in tumor cells is a positive prognostic indicator for postdiagnosis survival in patients with confirmed MPM. This is the first clinical study to correlate high TIF-2 expression with improved patient prognosis in any malignancy.

Membrane microdomain-associated uroplakin IIIa contributes to Src-dependent mechanisms of anti-apoptotic proliferation in human bladder carcinoma cells

Directory of Open Access Journals (Sweden)

Shigeru Kihira

2012-08-01

Our previous study demonstrated that tyrosine phosphorylation of p145met/β-subunit of hepatocyte growth factor receptor by epidermal growth factor receptor and Src contributes to the anti-apoptotic growth of human bladder carcinoma cell 5637 under serum-starved conditions. Here, we show that some other cell lines of human bladder carcinoma, but not other types of human cancer cells, also exhibit Src-dependent, anti-apoptotic proliferation under serum-starved conditions, and that low-density, detergent-insoluble membrane microdomains (MD serve as a structural platform for signaling events involving p145met, EGFR, and Src. As an MD-associated molecule that may contribute to bladder carcinoma-specific cellular function, we identified uroplakin IIIa (UPIIIa, an urothelium-specific protein. Results obtained so far revealed: 1 UPIIIa undergoes partial proteolysis in serum-starved cells; 2 a specific antibody to the extracellular domain of UPIIIa inhibits the proteolysis of UPIIIa and the activation of Src, and promotes apoptosis in serum-starved cells; and 3 knockdown of UPIIIa by short interfering RNA also promotes apoptosis in serum-starved cells. GM6001, a potent inhibitor of matrix metalloproteinase (MMP, inhibits the proteolysis of UPIIIa and promotes apoptosis in serum-starved cells. Furthermore, serum starvation promotes expression and secretion of the heparin-binding EGF-like growth factor in a manner that depends on the functions of MMP, Src, and UPIIIa. These results highlight a hitherto unknown signaling network involving a subset of MD-associated molecules in the anti-apoptotic mechanisms of human bladder carcinoma cells.

KAI1 suppresses HIF-1α and VEGF expression by blocking CDCP1-enhanced Src activation in prostate cancer

Directory of Open Access Journals (Sweden)

Park Jung-Jin

2012-03-01

Full Text Available Abstract Background KAI1 was initially identified as a metastasis-suppressor gene in prostate cancer. It is a member of the tetraspan transmembrane superfamily (TM4SF of membrane glycoproteins. As part of a tetraspanin-enriched microdomain (TEM, KAI1 inhibits tumor metastasis by negative regulation of Src. However, the underlying regulatory mechanism has not yet been fully elucidated. CUB-domain-containing protein 1 (CDCP1, which was previously known as tetraspanin-interacting protein in TEM, promoted metastasis via enhancement of Src activity. To better understand how KAI1 is involved in the negative regulation of Src, we here examined the function of KAI1 in CDCP1-mediated Src kinase activation and the consequences of this process, focusing on HIF-1 α and VEGF expression. Methods We used the human prostate cancer cell line PC3 which was devoid of KAI1 expression. Vector-transfected cells (PC3-GFP clone #8 and KAI1-expressing PC3 clones (PC3-KAI1 clone #5 and #6 were picked after stable transfection with KAI1 cDNA and selection in 800 μg/ml G418. Protein levels were assessed by immunoblotting and VEGF reporter gene activity was measured by assaying luciferase activitiy. We followed tumor growth in vivo and immunohistochemistry was performed for detection of HIF-1, CDCP1, and VHL protein level. Results We demonstrated that Hypoxia-inducible factor 1α (HIF-1α and VEGF expression were significantly inhibited by restoration of KAI1 in PC3 cells. In response to KAI1 expression, CDCP1-enhanced Src activation was down-regulated and the level of von Hippel-Lindau (VHL protein was significantly increased. In an in vivo xenograft model, KAI1 inhibited the expression of CDCP1 and HIF-1α. Conclusions These novel observations may indicate that KAI1 exerts profound metastasis-suppressor activity in the tumor malignancy process via inhibition of CDCP1-mediated Src activation, followed by VHL-induced HIF-1α degradation and, ultimately, decreased VEGF

76 FR 21404 - National Park Service Alaska Region's Subsistence Resource Commission (SRC) Program

Science.gov (United States)

2011-04-15

... National Park SRC will meet at the Shungnak Public School, 907-437-2151, in Shungnak, Alaska on Wednesday... changed, a notice will be published in local newspapers and announced on local radio stations prior to the...

Lipopolysaccharide induces VCAM-1 expression and neutrophil adhesion to human tracheal smooth muscle cells: Involvement of Src/EGFR/PI3-K/Akt pathway

International Nuclear Information System (INIS)

Lin, W.-N.; Luo, S.-F.; Wu, C.-B.; Lin, C.-C.; Yang, C.-M.

2008-01-01

In our previous study, LPS has been shown to induce vascular cell adhesion molecule-1(VCAM-1) expression through MAPKs and NF-κB in human tracheal smooth muscle cells (HTSMCs). In addition to these pathways, the non-receptor tyrosine kinases (Src), EGF receptor (EGFR), and phosphatidylinositol 3-kinase (PI3K) have been shown to be implicated in the expression of several inflammatory target proteins. Here, we reported that LPS-induced up-regulation of VCAM-1 enhanced the adhesion of neutrophils onto HTSMC monolayer, which was inhibited by LY294002 and wortmannin. LPS stimulated phosphorylation of protein tyrosine kinases including Src, PYK2, and EGFR, which were further confirmed using specific anti-phospho-Src, PYK2, or EGFR Ab, respectively, revealed by Western blotting. LPS-stimulated Src, PYK2, EGFR, and Akt phosphorylation and VCAM-1 expression were attenuated by the inhibitors of Src (PP1), EGFR (AG1478), PI3-K (LY294002 and wortmannin), and Akt (SH-5), respectively, or transfection with siRNAs of Src or Akt and shRNA of p110. LPS-induced VCAM-1 expression was also blocked by pretreatment with curcumin (a p300 inhibitor) or transfection with p300 siRNA. LPS-stimulated Akt activation translocated into nucleus and associated with p300 and VCAM-1 promoter region was further confirmed by immunofluorescence, immunoprecipitation, and chromatin immunoprecipitation assays. This association of Akt and p300 to VCAM-1 promoter was inhibited by pretreatment with PP1, AG1478, wortmannin, and SH-5. LPS-induced p300 activation enhanced VCAM-1 promoter activity and VCAM-1 mRNA expression. These results suggested that in HTSMCs, Akt phosphorylation mediated through transactivation of Src/PYK2/EGFR promoted the transcriptional p300 activity and eventually led to VCAM-1 expression induced by LPS

Role of c-Src inhibitor in the regulation of hepatocarcinoma cell ...

African Journals Online (AJOL)

SAM

2014-03-19

Mar 19, 2014 ... BEL-7402 cell line was used as HCC cell model for investigating the regulation of cell migration upon c-. Src inhibitors (PP2 and .... PDGF-BB were purchased from Enzo Life Sciences International,. USA; SU6656 Sigma (USA). .... Statistical analysis was done with Student's t-test for comparison of two ...

SRC-I solvent refined coal demonstration facility, Daviess County, Kentucky

Energy Technology Data Exchange (ETDEWEB)

1980-08-01

This volume of the Environmental Information Document on SRC-I contains appendices I-P. Information is provided for the preparation of the Environmental Impact Statement. Title listings of the appendices are: meteorology and air quality reports, December 1978 and March 1979; sound; economic, social, and cultural features; historical/architectural survey; archeological reports, 1979 and 1980; potential for burial and preservation of fossils; and alternate sites.

syk kinase activation by a src kinase-initiated activation loop phosphorylation chain reaction

Science.gov (United States)

El-Hillal, O.; Kurosaki, T.; Yamamura, H.; Kinet, J.-P.; Scharenberg, A. M.

1997-01-01

Activation of the syk tyrosine kinase occurs almost immediately following engagement of many types of antigen receptors, including Fc receptors, but the mechanism through which syk is activated is currently unclear. Here we demonstrate that Fc receptor-induced syk activation occurs as the result of phosphorylation of the syk activation loop by both src family kinases and other molecules of activated syk, suggesting that syk activation occurs as the result of a src kinase-initiated activation loop phosphorylation chain reaction. This type of activation mechanism predicts that syk activation would exhibit exponential kinetics, providing a potential explanation for its rapid and robust activation by even weak antigen receptor stimuli. We propose that a similar mechanism may be responsible for generating rapid activation of other cytoplasmic tyrosine kinases, such as those of the Bruton tyrosine kinase/tec family, as well. PMID:9050880

Fcγ-receptor IIa-mediated Src Signaling Pathway Is Essential for the Antibody-Dependent Enhancement of Ebola Virus Infection.

Directory of Open Access Journals (Sweden)

Wakako Furuyama

2016-12-01

Full Text Available Antibody-dependent enhancement (ADE of Ebola virus (EBOV infection has been demonstrated in vitro, raising concerns about the detrimental potential of some anti-EBOV antibodies. ADE has been described for many viruses and mostly depends on the cross-linking of virus-antibody complexes to cell surface Fc receptors, leading to enhanced infection. However, little is known about the molecular mechanisms underlying this phenomenon. Here we show that Fcγ-receptor IIa (FcγRIIa-mediated intracellular signaling through Src family protein tyrosine kinases (PTKs is required for ADE of EBOV infection. We found that deletion of the FcγRIIa cytoplasmic tail abolished EBOV ADE due to decreased virus uptake into cellular endosomes. Furthermore, EBOV ADE, but not non-ADE infection, was significantly reduced by inhibition of the Src family protein PTK pathway, which was also found to be important to promote phagocytosis/macropinocytosis for viral uptake into endosomes. We further confirmed a significant increase of the Src phosphorylation mediated by ADE. These data suggest that antibody-EBOV complexes bound to the cell surface FcγRIIa activate the Src signaling pathway that leads to enhanced viral entry into cells, providing a novel perspective for the general understanding of ADE of virus infection.

c-SRC mediates neurite outgrowth through recruitment of Crk to the scaffolding protein Sin/Efs without altering the kinetics of ERK activation

DEFF Research Database (Denmark)

Yang, Liang-Tung; Alexandropoulos, Konstantina; Sap, Jan

2002-01-01

moderate activation of endogenous SRC by receptor-protein-tyrosine phosphatase alpha (a physiological SRC activator). We show that such a qualitative change in the response to EGF is not accompanied by changes in the extent or kinetics of ERK induction in response to this factor. Instead, the pathway...

On locating steganographic payload using residuals

Science.gov (United States)

Quach, Tu-Thach

2011-02-01

Locating steganographic payload usingWeighted Stego-image (WS) residuals has been proven successful provided a large number of stego images are available. In this paper, we revisit this topic with two goals. First, we argue that it is a promising approach to locate payload by showing that in the ideal scenario where the cover images are available, the expected number of stego images needed to perfectly locate all load-carrying pixels is the logarithm of the payload size. Second, we generalize cover estimation to a maximum likelihood decoding problem and demonstrate that a second-order statistical cover model can be used to compute residuals to locate payload embedded by both LSB replacement and LSB matching steganography.

EFFECT OF COMMERCIALLY MANUFACTURED KAPPA SEMI-REFINED CARRAGEENAN (SRC WITH DIFFERENT PHOSPHATE SALTS ON YIELD, TEXTURAL AND SENSORY PROPERTIES OF BEEF MEAT

Directory of Open Access Journals (Sweden)

Marimuthu Murugesan

2016-06-01

Full Text Available The phosphate salts such as SDP, TSP, SHMP, DSP, TPPP, TKPP and STPP were blended with semi-refined kappa carrageenan (SRC individually and their aqueous solution was injected into the fresh beef meat. The highest weight gain (24% in fresh meat was obtained from injection of blend made with SRC and STPP as compared to other blends and control sample after forzen storage. Similarly, it also showed low cooking loss (24% with good testural and sensory proerties as compared to other blends. It can be concluded from the presnt study that blend of SRC and STPP can be used in beef meat for yield and quality improvement.

Inhibiting Src family tyrosine kinase activity blocks glutamate signalling to ERK1/2 and Akt/PKB but not JNK in cultured striatal neurones.

Science.gov (United States)

Crossthwaite, Andrew J; Valli, Haseeb; Williams, Robert J

2004-03-01

Glutamate receptor activation of mitogen-activated protein (MAP) kinase signalling cascades has been implicated in diverse neuronal functions such as synaptic plasticity, development and excitotoxicity. We have previously shown that Ca2+-influx through NMDA receptors in cultured striatal neurones mediates the phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2) and Akt/protein kinase B (PKB) through a phosphatidylinositol 3-kinase (PI 3-kinase)-dependent pathway. Exposing neurones to the Src family tyrosine kinase inhibitor PP2, but not the inactive analogue PP3, inhibited NMDA receptor-induced phosphorylation of ERK1/2 and Akt/PKB in a concentration-dependent manner, and reduced cAMP response element-binding protein (CREB) phosphorylation. To establish a link between Src family tyrosine kinase-mediated phosphorylation and PI 3-kinase signalling, affinity precipitation experiments were performed with the SH2 domains of the PI 3-kinase regulatory subunit p85. This revealed a Src-dependent phosphorylation of a focal adhesion kinase (FAK)-p85 complex on glutamate stimulation. Demonstrating that PI3-kinase is not ubiquitously involved in NMDA receptor signal transduction, the PI 3-kinase inhibitors wortmannin and LY294002 did not prevent NMDA receptor Ca2+-dependent phosphorylation of c-Jun N-terminal kinase 1/2 (JNK1/2). Further, inhibiting Src family kinases increased NMDA receptor-dependent JNK1/2 phosphorylation, suggesting that Src family kinase-dependent cascades may physiologically limit signalling to JNK. These results demonstrate that Src family tyrosine kinases and PI3-kinase are pivotal regulators of NMDA receptor signalling to ERK/Akt and JNK in striatal neurones.

Rapid activation of Rac GTPase in living cells by force is independent of Src.

Directory of Open Access Journals (Sweden)

Yeh-Chuin Poh

2009-11-01

Full Text Available It is well known that mechanical forces are crucial in regulating functions of every tissue and organ in a human body. However, it remains unclear how mechanical forces are transduced into biochemical activities and biological responses at the cellular and molecular level. Using the magnetic twisting cytometry technique, we applied local mechanical stresses to living human airway smooth muscle cells with a magnetic bead bound to the cell surface via transmembrane adhesion molecule integrins. The temporal and spatial activation of Rac, a small guanosine triphosphatase, was quantified using a fluorescent resonance energy transfer (FRET method that measures changes in Rac activity in response to mechanical stresses by quantifying intensity ratios of ECFP (enhanced cyan fluorescent protein as a donor and YPet (a variant yellow fluorescent protein as an acceptor of the Rac biosensor. The applied stress induced rapid activation (less than 300 ms of Rac at the cell periphery. In contrast, platelet derived growth factor (PDGF induced Rac activation at a much later time (>30 sec. There was no stress-induced Rac activation when a mutant form of the Rac biosensor (RacN17 was transfected or when the magnetic bead was coated with transferrin or with poly-L-lysine. It is known that PDGF-induced Rac activation depends on Src activity. Surprisingly, pre-treatment of the cells with specific Src inhibitor PP1 or knocking-out Src gene had no effects on stress-induced Rac activation. In addition, eliminating lipid rafts through extraction of cholesterol from the plasma membrane did not prevent stress-induced Rac activation, suggesting a raft-independent mechanism in governing the Rac activation upon mechanical stimulation. Further evidence indicates that Rac activation by stress depends on the magnitudes of the applied stress and cytoskeletal integrity. Our results suggest that Rac activation by mechanical forces is rapid, direct and does not depend on Src

Fisetin suppresses malignant proliferation in human oral squamous cell carcinoma through inhibition of Met/Src signaling pathways.

Science.gov (United States)

Li, Yan-Shu; Qin, Xing-Jun; Dai, Wei

2017-01-01

Fisetin (3,7,3',4'-tetrahydroxyflavone) is a dietary flavonoid and has been indicated as a novel anti-cancer agent in several types of cancer cells. However, the mechanisms underlying the effect of fisetin in human oral squamous cell carcinoma (OSCC) remain unclear. Here, we report that fisetin significantly inhibits tumor cell proliferation and induces apoptosis in OSCC (UM-SCC-23 and Tca-8113) cancer cell lines. Further analysis demonstrates that fisetin also inhibits Met/Src signaling pathways using the PathScan ® receptor tyrosine kinases (RTK) Signaling Antibody Array Kit. Fisetin resulted in decreased basal expression of Met and Src protein in UM-SCC-23 cancer cell lines, which validated by western blot. A student's t -test (two-tailed) was used to compare differences between groups. Furthermore, fisetin significantly inhibited the expression of a disintegrin and metalloproteinase 9 (ADAM9) protein in OSCC cells. Taken together, these results provide novel insights into the mechanism of fisetin and suggest potential therapeutic strategies for human OSCC by blocking the Met/Src signaling pathways.

Technical note: Equivalent genomic models with a residual polygenic effect.

Science.gov (United States)

Liu, Z; Goddard, M E; Hayes, B J; Reinhardt, F; Reents, R

2016-03-01

Routine genomic evaluations in animal breeding are usually based on either a BLUP with genomic relationship matrix (GBLUP) or single nucleotide polymorphism (SNP) BLUP model. For a multi-step genomic evaluation, these 2 alternative genomic models were proven to give equivalent predictions for genomic reference animals. The model equivalence was verified also for young genotyped animals without phenotypes. Due to incomplete linkage disequilibrium of SNP markers to genes or causal mutations responsible for genetic inheritance of quantitative traits, SNP markers cannot explain all the genetic variance. A residual polygenic effect is normally fitted in the genomic model to account for the incomplete linkage disequilibrium. In this study, we start by showing the proof that the multi-step GBLUP and SNP BLUP models are equivalent for the reference animals, when they have a residual polygenic effect included. Second, the equivalence of both multi-step genomic models with a residual polygenic effect was also verified for young genotyped animals without phenotypes. Additionally, we derived formulas to convert genomic estimated breeding values of the GBLUP model to its components, direct genomic values and residual polygenic effect. Third, we made a proof that the equivalence of these 2 genomic models with a residual polygenic effect holds also for single-step genomic evaluation. Both the single-step GBLUP and SNP BLUP models lead to equal prediction for genotyped animals with phenotypes (e.g., reference animals), as well as for (young) genotyped animals without phenotypes. Finally, these 2 single-step genomic models with a residual polygenic effect were proven to be equivalent for estimation of SNP effects, too. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Development and experimental test of support vector machines virtual screening method for searching Src inhibitors from large compound libraries

Directory of Open Access Journals (Sweden)

Han Bucong

2012-11-01

Full Text Available Abstract Background Src plays various roles in tumour progression, invasion, metastasis, angiogenesis and survival. It is one of the multiple targets of multi-target kinase inhibitors in clinical uses and trials for the treatment of leukemia and other cancers. These successes and appearances of drug resistance in some patients have raised significant interest and efforts in discovering new Src inhibitors. Various in-silico methods have been used in some of these efforts. It is desirable to explore additional in-silico methods, particularly those capable of searching large compound libraries at high yields and reduced false-hit rates. Results We evaluated support vector machines (SVM as virtual screening tools for searching Src inhibitors from large compound libraries. SVM trained and tested by 1,703 inhibitors and 63,318 putative non-inhibitors correctly identified 93.53%~ 95.01% inhibitors and 99.81%~ 99.90% non-inhibitors in 5-fold cross validation studies. SVM trained by 1,703 inhibitors reported before 2011 and 63,318 putative non-inhibitors correctly identified 70.45% of the 44 inhibitors reported since 2011, and predicted as inhibitors 44,843 (0.33% of 13.56M PubChem, 1,496 (0.89% of 168 K MDDR, and 719 (7.73% of 9,305 MDDR compounds similar to the known inhibitors. Conclusions SVM showed comparable yield and reduced false hit rates in searching large compound libraries compared to the similarity-based and other machine-learning VS methods developed from the same set of training compounds and molecular descriptors. We tested three virtual hits of the same novel scaffold from in-house chemical libraries not reported as Src inhibitor, one of which showed moderate activity. SVM may be potentially explored for searching Src inhibitors from large compound libraries at low false-hit rates.

Development and experimental test of support vector machines virtual screening method for searching Src inhibitors from large compound libraries.

Science.gov (United States)

Han, Bucong; Ma, Xiaohua; Zhao, Ruiying; Zhang, Jingxian; Wei, Xiaona; Liu, Xianghui; Liu, Xin; Zhang, Cunlong; Tan, Chunyan; Jiang, Yuyang; Chen, Yuzong

2012-11-23

Src plays various roles in tumour progression, invasion, metastasis, angiogenesis and survival. It is one of the multiple targets of multi-target kinase inhibitors in clinical uses and trials for the treatment of leukemia and other cancers. These successes and appearances of drug resistance in some patients have raised significant interest and efforts in discovering new Src inhibitors. Various in-silico methods have been used in some of these efforts. It is desirable to explore additional in-silico methods, particularly those capable of searching large compound libraries at high yields and reduced false-hit rates. We evaluated support vector machines (SVM) as virtual screening tools for searching Src inhibitors from large compound libraries. SVM trained and tested by 1,703 inhibitors and 63,318 putative non-inhibitors correctly identified 93.53%~ 95.01% inhibitors and 99.81%~ 99.90% non-inhibitors in 5-fold cross validation studies. SVM trained by 1,703 inhibitors reported before 2011 and 63,318 putative non-inhibitors correctly identified 70.45% of the 44 inhibitors reported since 2011, and predicted as inhibitors 44,843 (0.33%) of 13.56M PubChem, 1,496 (0.89%) of 168 K MDDR, and 719 (7.73%) of 9,305 MDDR compounds similar to the known inhibitors. SVM showed comparable yield and reduced false hit rates in searching large compound libraries compared to the similarity-based and other machine-learning VS methods developed from the same set of training compounds and molecular descriptors. We tested three virtual hits of the same novel scaffold from in-house chemical libraries not reported as Src inhibitor, one of which showed moderate activity. SVM may be potentially explored for searching Src inhibitors from large compound libraries at low false-hit rates.

Solvent Refined Coal (SRC) process: trace elements. Research and development report No. 53, interim report No. 31, August 1976-July 1977. Volume III. Pilot plant development work. Part 6

International Nuclear Information System (INIS)

1980-03-01

Results are presented on a study of the distribution and fate of 34 trace elements in the Solvent Refined Coal Process at the pilot plant located at Fort Lewis, Washington, and operated by The Pittsburg and Midway Coal Mining Co. under contract with the US Department of Energy. Neutron activation analysis was used to determine Ti, V, Ca, Mg, Al, Cl, Mn, As, Sb, Se, Hg, Br, Co, Ni, Cr, Fe, Na, Rb, Cs, K, Sc, Tb, Eu, Sm, Ce, La, Sr, Ba, Th, Hf, Ta, Ga, Zr, and Cu in feed coals, process solvent, Solvent Refined Coal (SRC), mineral residues, wet filter cake, by-product solvents, process and effluent waters and by-product sulfur. The sample points were chosen such that the major process streams were adequately described and that the major input and output materials were included. Atomic absorption spectrophotometry was used to measure the toxic elements Pb, Cd, Be in plant-derived solvents, effluent water and Hamer Marsh water. Specific methods were developed for analysis of a wide range of material compositions. The neutron activation analysis procedures were divided into short and long irradiation procedures for elements with short half lives (less than 3 hours) and intermediate to long half lives ( 8 hours to 5.2 years). Data are presented for a third equilibrium set of samples from the SRC-I process and compared to two similar sets analyzed previously. A material balance (or budget) was calculated for each element from the concentration data and the yields of each process fraction. Data are also presented on a study of carbon monoxide addition to the hydrogen stream and its effect on trace elements, and trace element data on a study of thirty-six plant effluent water samples taken during an SRC-I production run

Integrin αβ1, αvβ, α6β effectors p130Cas, Src and talin regulate carcinoma invasion and chemoresistance

International Nuclear Information System (INIS)

Sansing, Hope A.; Sarkeshik, Ali; Yates, John R.; Patel, Vyomesh; Gutkind, J. Silvio; Yamada, Kenneth M.; Berrier, Allison L.

2011-01-01

Research highlights: → Proteomics of clustered integrin αβ1, α v β, α 6 β receptors in oral carcinoma. → p130Cas, Dek, Src and talin regulate oral carcinoma invasion. → p130Cas, talin, Src and zyxin regulate oral carcinoma resistance to cisplatin. -- Abstract: Ligand engagement by integrins induces receptor clustering and formation of complexes at the integrin cytoplasmic face that controls cell signaling and cytoskeletal dynamics critical for adhesion-dependent processes. This study searches for a subset of integrin effectors that coordinates both tumor cell invasion and resistance to the chemotherapeutic drug cisplatin in oral carcinomas. Candidate integrin effectors were identified in a proteomics screen of proteins recruited to clustered integrin αβ1, α v β or α 6 β receptors in oral carcinomas. Proteins with diverse functions including microtubule and actin binding proteins, and factors involved in trafficking, transcription and translation were identified in oral carcinoma integrin complexes. Knockdown of effectors in the oral carcinoma HN12 cells revealed that p130Cas, Dek, Src and talin were required for invasion through Matrigel. Disruption of talin or p130Cas by RNA interference increased resistance to cisplatin, whereas targeting Dek, Src or zyxin reduced HN12 resistance to cisplatin. Analysis of the spreading of HN12 cells on collagen I and laminin I revealed that a decrease in p130Cas or talin expression inhibited spreading on both matrices. Interestingly, a reduction in zyxin expression enhanced spreading on laminin I and inhibited spreading on collagen I. Reduction of Dek, Src, talin or zyxin expression reduced HN12 proliferation by 30%. Proliferation was not affected by a reduction in p130Cas expression. We conclude that p130Cas, Src and talin function in both oral carcinoma invasion and resistance to cisplatin.

Part I---Evaluating Effects of Oligomer Formation on Cytochrome P450 2C9 Electron Transfer and Drug Metabolism, Part II---Utilizing Molecular Modeling Techniques to Study the Src-Interacting Proteins Actin Filament Associated Protein of 110 kDa (AFAP-110) and Cortactin

Science.gov (United States)

Jett, John Edward, Jr.

nanopillars, the immobilization of CYP2C9 enzymes to those nanopillars, and the utilization of the array to perform conductive probe atomic force microscopy experiments examining the electron transfer process of CYP2C9 in the absence and presence of substrate molecules. Part II. The Src protein has been known to play a role in cancer cell progression for over 30 years. The function of a non-receptor tyrosine kinase such as Src is to relay extracellular signals through intracellular tyrosine phosphorylation. As a tyrosine kinase, Src and the cellular signaling pathways it is involved in play many functional roles in the cell, both in cellular proliferation and in cytoskeletal dynamics, cell adhesion, motility and invasion. Two of the many proteins comprising Src cellular signaling pathways are actin filament associated protein of 110 kDa (AFAP-110) and cortactin. AFAP-110 is a known activator of Src; one mechanism to abrogate the AFAP-110-induced activation of Src is to inhibit their colocalization within the cell. This colocalization is expected to occur when the pleckstrin homology (PH1 and PH2) domains of AFAP-110 are allowed to interact with membrane-bound phospholipids. Cortactin, on the other hand, is a cytosolic protein capable of being phosphorylated on various tyrosine residues, activating it and allowing it to interact with actin. The Src homology 2 (SH2) domain of Src has been shown to be capable of interacting with cortactin, an association which will be probed here. This section of the dissertation will discuss the use of molecular modeling techniques to develop structural models of the AFAP-110 PH1 and PH2 domains and use them to make predictions about how the protein interacts with phospholipids in the plasma membrane and how they might be stabilized to interact with other proteins. Structural models were designed using homology modeling methods, docking programs were used to predict key residues of AFAP-110 involved in binding to phospholipids and mutational

Characterization of cortactin as an in vivo protein kinase D substrate: interdependence of sites and potentiation by Src.

Science.gov (United States)

De Kimpe, Line; Janssens, Katrien; Derua, Rita; Armacki, Milena; Goicoechea, Silvia; Otey, Carol; Waelkens, Etienne; Vandoninck, Sandy; Vandenheede, Jackie R; Seufferlein, Thomas; Van Lint, Johan

2009-02-01

Protein Kinase D (PKD) has been implicated in the regulation of actin turnover at the leading edge, invasion and migration. In particular, a complex between cortactin, paxillin and PKD in the invadopodia of invasive breast cancer cells has been described earlier, but so far this complex remained ill defined. Here we have investigated the possible role of PKD as a cortactin kinase. Using a mass spectrometric approach, we found that PKD phosphorylates cortactin on Ser 298 in the 6th cortactin repeat region and on Ser 348, right before the helical-proline rich domain of cortactin. We developed phosphospecific antibodies against these phosphorylated sequences, and used them as tools to follow the in vivo phosphorylation of cortactin by PKD. Examination of cortactin phosphorylation kinetics revealed that Ser 298 serves as a priming site for subsequent phosphorylation of Ser 348. Src, a well-known cortactin kinase, strongly potentiated the in vivo PKD mediated cortactin phosphorylation. This Src effect is neither mediated by pre-phosphorylation of cortactin nor by activation of PKD by Src. Phosphorylation of cortactin by PKD does not affect its subcellular localization, nor does it affect its translocation to podosomes or membrane ruffles. Moreover, there was no effect of PKD mediated cortactin phosphorylation on EGF receptor degradation and LPA induced migration. Taken together, these data establish cortactin as a novel PKD substrate and reveal a novel connection between Src and PKD.

Structure of the interleukin-2 tyrosine kinase Src homology 2 domain; comparison between X-ray and NMR-derived structures

International Nuclear Information System (INIS)

Joseph, Raji E.; Ginder, Nathaniel D.; Hoy, Julie A.; Nix, Jay C.; Fulton, D. Bruce; Honzatko, Richard B.; Andreotti, Amy H.

2012-01-01

The interleukin-2 tyrosine kinase Src homology 2 domain was crystallized and its structure was solved to 2.35 Å resolution. The structure reveals a domain-swapped dimer that is related to other dimeric SH2 domains solved previously. The cis–trans-prolyl isomerization that is evident from solution studies of Itk SH2 cannot be observed in the crystal structure. The crystal structure of the interleukin-2 tyrosine kinase Src homology domain (Itk SH2) is described and it is found that unlike in studies of this domain using NMR spectroscopy, cis–trans-prolyl isomerization is not readily detected in the crystal structure. Based on similarities between the Itk SH2 crystal form and the cis form of the Itk SH2 NMR structure, it is concluded that it is likely that the prolyl imide bond at least in part adopts the cis conformation in the crystal form. However, the lack of high-resolution data and the dynamic nature of the proline-containing loop mean that the precise imide-bond conformation cannot be determined and prolyl cis–trans isomerization in the crystal cannot be ruled out. Given the preponderance of structures that have been solved by X-ray crystallography in the Protein Data Bank, this result supports the notion that prolyl isomerization in folded proteins has been underestimated among known structures. Interestingly, while the precise status of the proline residue is ambiguous, Itk SH2 crystallizes as a domain-swapped dimer. The domain-swapped structure of Itk SH2 is similar to the domain-swapped SH2 domains of Grb2 and Nck, with domain swapping occurring at the β-meander region of all three SH2 domains. Thus, for Itk SH2 structural analysis by NMR spectroscopy and X-ray crystallography revealed very different structural features: proline isomerization versus domain-swapped dimerization, respectively

Inactivation of Src-to-Ezrin Pathway: A Possible Mechanism in the Ouabain-Mediated Inhibition of A549 Cell Migration

Directory of Open Access Journals (Sweden)

Hye Kyoung Shin

2015-01-01

Full Text Available Ouabain, a cardiac glycoside found in plants, is primarily used in the treatment of congestive heart failure and arrhythmia because of its ability to inhibit Na+/K+-ATPase pump. Recently ouabain has been shown to exert anticancer effects but the underlying mechanism is not clear. Here, we explored the molecular mechanism by which ouabain exerts anticancer effects in human lung adenocarcinoma. Employing proteomic techniques, we found 7 proteins downregulated by ouabain in A549 including p-ezrin, a protein associated with pulmonary cancer metastasis in a dose-dependent manner. In addition, when the relative phosphorylation levels of 39 intracellular proteins were compared between control and ouabain-treated A549 cells, p-Src (Y416 was also found to be downregulated by ouabain. Furthermore, western blot revealed the ouabain-mediated downregulation of p-FAK (Y925, p-paxillin (Y118, p130CAS, and Na+/K+-ATPase subunits that have been shown to be involved in the migration of cancer cells. The inhibitory effect of ouabain and Src inhibitor PP2 on the migration of A549 cells was confirmed by Boyden chamber assay. Anticancer effects of ouabain in A549 cells appear to be related to its ability to regulate and inactivate Src-to-ezrin signaling, and proteins involved in focal adhesion such as Src, FAK, and p130CAS axis are proposed here.

Barium promotes anchorage-independent growth and invasion of human HaCaT keratinocytes via activation of c-SRC kinase.

Science.gov (United States)

Thang, Nguyen Dinh; Yajima, Ichiro; Kumasaka, Mayuko Y; Ohnuma, Shoko; Yanagishita, Takeshi; Hayashi, Rumiko; Shekhar, Hossain U; Watanabe, Daisuke; Kato, Masashi

2011-01-01

Explosive increases in skin cancers have been reported in more than 36 million patients with arsenicosis caused by drinking arsenic-polluted well water. This study and previous studies showed high levels of barium as well as arsenic in the well water. However, there have been no reports showing a correlation between barium and cancer. In this study, we examined whether barium (BaCl(2)) may independently have cancer-related effects on human precancerous keratinocytes (HaCaT). Barium (5-50 µM) biologically promoted anchorage-independent growth and invasion of HaCaT cells in vitro. Barium (5 µM) biochemically enhanced activities of c-SRC, FAK, ERK and MT1-MMP molecules, which regulate anchorage-independent growth and/or invasion. A SRC kinase specific inhibitor, protein phosphatase 2 (PP2), blocked barium-mediated promotion of anchorage-independent growth and invasion with decreased c-SRC kinase activity. Barium (2.5-5 µM) also promoted anchorage-independent growth and invasion of fibroblasts (NIH3T3) and immortalized nontumorigenic melanocytes (melan-a), but not transformed cutaneous squamous cell carcinoma (HSC5 and A431) and malignant melanoma (Mel-ret) cells, with activation of c-SRC kinase. Taken together, our biological and biochemical findings newly suggest that the levels of barium shown in drinking well water independently has the cancer-promoting effects on precancerous keratinocytes, fibroblast and melanocytes in vitro.

Cysteine residues 244 and 458–459 within the catalytic subunit of Na,K-ATPase control the enzyme's hydrolytic and signaling function under hypoxic conditions

Directory of Open Access Journals (Sweden)

Irina Yu. Petrushanko

2017-10-01

Full Text Available Our previous findings suggested that reversible thiol modifications of cysteine residues within the actuator (AD and nucleotide binding domain (NBD of the Na,K-ATPase may represent a powerful regulatory mechanism conveying redox- and oxygen-sensitivity of this multifunctional enzyme. S-glutathionylation of Cys244 in the AD and Cys 454-458-459 in the NBD inhibited the enzyme and protected cysteines’ thiol groups from irreversible oxidation under hypoxic conditions. In this study mutagenesis approach was used to assess the role these cysteines play in regulation of the Na,K-ATPase hydrolytic and signaling functions. Several constructs of mouse α1 subunit of the Na,K-ATPase were produced in which Cys244, Cys 454-458-459 or Cys 244-454-458-459 were replaced by alanine. These constructs were expressed in human HEK293 cells. Non-transfected cells and those expressing murine α1 subunit were exposed to hypoxia or treated with oxidized glutathione (GSSG. Both conditions induced inhibition of the wild type Na,K-ATPase. Enzymes containing mutated mouse α1 lacking Cys244 or all four cysteines (Cys 244-454-458-459 were insensitive to hypoxia. Inhibitory effect of GSSG was observed for wild type murine Na,K-ATPase, but was less pronounced in Cys454-458-459Ala mutant and completely absent in the Cys244Ala and Cys 244-454-458-459Ala mutants. In cells, expressing wild type enzyme, ouabain induced activation of Src and Erk kinases under normoxic conditions, whereas under hypoxic conditions this effect was inversed. Cys454-458-459Ala substitution abolished Src kinase activation in response to ouabain treatment, uncoupled Src from Erk signaling, and interfered with O2-sensitivity of Na,K-ATPase signaling function. Moreover, modeling predicted that S-glutathionylation of Cys 458 and 459 should prevent inhibitory binding of Src to NBD. Our data indicate for the first time that cysteine residues within the AD and NBD influence hydrolytic as well as receptor

Multiple Kernel Sparse Representation based Orthogonal Discriminative Projection and Its Cost-Sensitive Extension.

Science.gov (United States)

Zhang, Guoqing; Sun, Huaijiang; Xia, Guiyu; Sun, Quansen

2016-07-07

Sparse representation based classification (SRC) has been developed and shown great potential for real-world application. Based on SRC, Yang et al. [10] devised a SRC steered discriminative projection (SRC-DP) method. However, as a linear algorithm, SRC-DP cannot handle the data with highly nonlinear distribution. Kernel sparse representation-based classifier (KSRC) is a non-linear extension of SRC and can remedy the drawback of SRC. KSRC requires the use of a predetermined kernel function and selection of the kernel function and its parameters is difficult. Recently, multiple kernel learning for SRC (MKL-SRC) [22] has been proposed to learn a kernel from a set of base kernels. However, MKL-SRC only considers the within-class reconstruction residual while ignoring the between-class relationship, when learning the kernel weights. In this paper, we propose a novel multiple kernel sparse representation-based classifier (MKSRC), and then we use it as a criterion to design a multiple kernel sparse representation based orthogonal discriminative projection method (MK-SR-ODP). The proposed algorithm aims at learning a projection matrix and a corresponding kernel from the given base kernels such that in the low dimension subspace the between-class reconstruction residual is maximized and the within-class reconstruction residual is minimized. Furthermore, to achieve a minimum overall loss by performing recognition in the learned low-dimensional subspace, we introduce cost information into the dimensionality reduction method. The solutions for the proposed method can be efficiently found based on trace ratio optimization method [33]. Extensive experimental results demonstrate the superiority of the proposed algorithm when compared with the state-of-the-art methods.

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

Estrogen Receptor Folding Modulates cSrc Kinase SH2 Interaction via a Helical Binding Mode.

Science.gov (United States)

Nieto, Lidia; Tharun, Inga M; Balk, Mark; Wienk, Hans; Boelens, Rolf; Ottmann, Christian; Milroy, Lech-Gustav; Brunsveld, Luc

2015-11-20

The estrogen receptors (ERs) feature, next to their transcriptional role, important nongenomic signaling actions, with emerging clinical relevance. The Src Homology 2 (SH2) domain mediated interaction between cSrc kinase and ER plays a key role in this; however the molecular determinants of this interaction have not been elucidated. Here, we used phosphorylated ER peptide and semisynthetic protein constructs in a combined biochemical and structural study to, for the first time, provide a quantitative and structural characterization of the cSrc SH2-ER interaction. Fluorescence polarization experiments delineated the SH2 binding motif in the ER sequence. Chemical shift perturbation analysis by nuclear magnetic resonance (NMR) together with molecular dynamics (MD) simulations allowed us to put forward a 3D model of the ER-SH2 interaction. The structural basis of this protein-protein interaction has been compared with that of the high affinity SH2 binding sequence GpYEEI. The ER features a different binding mode from that of the "two-pronged plug two-hole socket" model in the so-called specificity determining region. This alternative binding mode is modulated via the folding of ER helix 12, a structural element directly C-terminal of the key phosphorylated tyrosine. The present findings provide novel molecular entries for understanding nongenomic ER signaling and targeting the corresponding disease states.

Activity of dual SRC-ABL inhibitors highlights the role of BCR/ABL kinase dynamics in drug resistance

Science.gov (United States)

Azam, Mohammad; Nardi, Valentina; Shakespeare, William C.; Metcalf, Chester A.; Bohacek, Regine S.; Wang, Yihan; Sundaramoorthi, Raji; Sliz, Piotr; Veach, Darren R.; Bornmann, William G.; Clarkson, Bayard; Dalgarno, David C.; Sawyer, Tomi K.; Daley, George Q.

2006-01-01

Mutation in the ABL kinase domain is the principal mechanism of imatinib resistance in patients with chronic myelogenous leukemia. Many mutations favor active kinase conformations that preclude imatinib binding. Because the active forms of ABL and SRC resemble one another, we tested two dual SRC-ABL kinase inhibitors, AP23464 and PD166326, against 58 imatinib-resistant (IMR) BCR/ABL kinase variants. Both compounds potently inhibit most IMR variants, and in vitro drug selection demonstrates that active (AP23464) and open (PD166326) conformation-specific compounds are less susceptible to resistance than imatinib. Combinations of inhibitors suppressed essentially all resistance mutations, with the notable exception of T315I. Guided by mutagenesis studies and molecular modeling, we designed a series of AP23464 analogues to target T315I. The analogue AP23846 inhibited both native and T315I variants of BCR/ABL with submicromolar potency but showed nonspecific cellular toxicity. Our data illustrate how conformational dynamics of the ABL kinase accounts for the activity of dual SRC-ABL inhibitors against IMR-mutants and provides a rationale for combining conformation specific inhibitors to suppress resistance. PMID:16754879

Na,K-ATPase activity modulates Src activation: A role for ATP/ADP ratio.

NARCIS (Netherlands)

Weigand, K.M.; Swarts, H.G.P.; Fedosova, N.U.; Russel, F.G.M.; Koenderink, J.B.

2012-01-01

Digitalis-like compounds (DLCs), specific inhibitors of Na,K-ATPase, are implicated in cellular signaling. Exposure of cell cultures to ouabain, a well-known DLC, leads to up- or down regulation of various processes and involves activation of Src kinase. Since Na,K-ATPase is the only known target

Nuclear expression of Lyn, a Src family kinase member, is associated with poor prognosis in renal cancer patients.

Science.gov (United States)

Roseweir, Antonia K; Qayyum, Tahir; Lim, Zhi; Hammond, Rachel; MacDonald, Alasdair I; Fraser, Sioban; Oades, Grenville M; Aitchison, Michael; Jones, Robert J; Edwards, Joanne

2016-03-16

8000 cases of renal cancer are diagnosed each year in the UK, with a five-year survival rate of 50%. Treatment options are limited; a potential therapeutic target is the Src family kinases (SFKs). SFKs have roles in multiple oncogenic processes and promote metastases in solid tumours. The aim of this study was to investigate SFKs as potential therapeutic targets for clear cell renal cell carcinoma (ccRCC). SFKs expression was assessed in a tissue microarray consisting of 192 ccRCC patients with full clinical follow-up. SFK inhibitors, dasatinib and saracatinib, were assessed in early ccRCC cell lines, 786-O and 769-P and a metastatic ccRCC cell line, ACHN (± Src) for effects on protein expression, apoptosis, proliferation and wound healing. High nuclear expression of Lyn and the downstream marker of activation, paxillin, were associated with decreased patient survival. Conversely, high cytoplasmic expression of other SFK members and downstream marker of activation, focal adhesion kinase (FAK) were associated with increased patient survival. Treatment of non-metastatic 786-O and 769-P cells with dasatinib, dose dependently reduced SFK activation, shown via SFK (Y(419)) and FAK (Y(861)) phosphorylation, with no effect in metastatic ACHN cells. Dasatinib also increased apoptosis, while decreasing proliferation and migration in 786-O and 769-P cell lines, both in the presence and absence of Src protein. Our data suggests that nuclear Lyn is a potential therapeutic target for ccRCC and dasatinib affects cellular functions associated with cancer progression via a Src kinase independent mechanism.

FAK/src-family dependent activation of the Ste20-like kinase SLK is required for microtubule-dependent focal adhesion turnover and cell migration.

Directory of Open Access Journals (Sweden)

Simona Wagner

2008-04-01

Full Text Available Cell migration involves a multitude of signals that converge on cytoskeletal reorganization, essential for development, immune responses and tissue repair. Using knockdown and dominant negative approaches, we show that the microtubule-associated Ste20-like kinase SLK is required for focal adhesion turnover and cell migration downstream of the FAK/c-src complex. Our results show that SLK co-localizes with paxillin, Rac1 and the microtubules at the leading edge of migrating cells and is activated by scratch wounding. SLK activation is dependent on FAK/c-src/MAPK signaling, whereas SLK recruitment to the leading edge is src-dependent but FAK independent. Our results show that SLK represents a novel focal adhesion disassembly signal.

Recording Process Documentation for Provenance

NARCIS (Netherlands)

Groth, P.T.; Moreau, L

2009-01-01

Scientific and business communities are adopting large-scale distributed systems as a means to solve a wide range of resource-intensive tasks. These communities also have requirements in terms of provenance. We define the provenance of a result produced by a distributed system as the process that

Effects of Src Kinase Inhibition on Expression of Protein Tyrosine Phosphatase 1B after Brain Hypoxia in a Piglet Animal Model

Directory of Open Access Journals (Sweden)

Dimitrios Angelis

2017-01-01

Full Text Available Background. Protein tyrosine phosphatases (PTPs in conjunction with protein tyrosine kinases (PTKs regulate cellular processes by posttranslational modifications of signal transduction proteins. PTP nonreceptor type 1B (PTP-1B is an enzyme of the PTP family. We have previously shown that hypoxia induces an increase in activation of a class of nonreceptor PTK, the Src kinases. In the present study, we investigated the changes that occur in the expression of PTP-1B in the cytosolic component of the brain of newborn piglets acutely after hypoxia as well as long term for up to 2 weeks. Methods. Newborn piglets were divided into groups: normoxia, hypoxia, hypoxia followed by 1 day and 15 days in FiO2 0.21, and hypoxia pretreated with Src kinase inhibitor PP2, prior to hypoxia followed by 1 day and 15 days. Hypoxia was achieved by providing 7% FiO2 for 1 hour and PTP-1B expression was measured via immunoblotting. Results. PTP-1B increased posthypoxia by about 30% and persisted for 2 weeks while Src kinase inhibition attenuated the expected PTP-1B-increased expression. Conclusions. Our study suggests that Src kinase mediates a hypoxia-induced increased PTP-1B expression.

STEP activation by Gαq coupled GPCRs opposes Src regulation of NMDA receptors containing the GluN2A subunit

Science.gov (United States)

Tian, Meng; Xu, Jian; Lei, Gang; Lombroso, Paul J.; Jackson, Michael F.; MacDonald, John F.

2016-01-01

N-methyl-D-aspartate receptors (NMDARs) are necessary for the induction of synaptic plasticity and for the consolidation of learning and memory. NMDAR function is tightly regulated by functionally opposed families of kinases and phosphatases. Herein we show that the striatal-enriched protein tyrosine phosphatase (STEP) is recruited by Gαq-coupled receptors, including the M1 muscarinic acetylcholine receptor (M1R), and opposes the Src tyrosine kinase-mediated increase in the function of NMDARs composed of GluN2A. STEP activation by M1R stimulation requires IP3Rs and can depress NMDA-evoked currents with modest intracellular Ca2+ buffering. Src recruitment by M1R stimulation requires coincident NMDAR activation and can augment NMDA-evoked currents with high intracellular Ca2+ buffering. Our findings suggest that Src and STEP recruitment is contingent on differing intracellular Ca2+ dynamics that dictate whether NMDAR function is augmented or depressed following M1R stimulation. PMID:27857196

Integrin {alpha}{beta}1, {alpha}{sub v}{beta}, {alpha}{sub 6}{beta} effectors p130Cas, Src and talin regulate carcinoma invasion and chemoresistance

Energy Technology Data Exchange (ETDEWEB)

Sansing, Hope A. [Department of Oral and Craniofacial Biology, Louisiana State University Health Sciences Center-New Orleans, School of Dentistry, New Orleans, LA (United States); Sarkeshik, Ali; Yates, John R. [Department of Chemical Physiology, Scripps Research Institute, La Jolla, CA (United States); Patel, Vyomesh; Gutkind, J. Silvio [Oral and Pharyngeal Cancer Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD (United States); Yamada, Kenneth M. [Laboratory of Cell and Developmental Biology, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD (United States); Berrier, Allison L., E-mail: allison.berrier@gmail.com [Department of Oral and Craniofacial Biology, Louisiana State University Health Sciences Center-New Orleans, School of Dentistry, New Orleans, LA (United States)

2011-03-11

Research highlights: {yields} Proteomics of clustered integrin {alpha}{beta}1, {alpha}{sub v}{beta}, {alpha}{sub 6}{beta} receptors in oral carcinoma. {yields} p130Cas, Dek, Src and talin regulate oral carcinoma invasion. {yields} p130Cas, talin, Src and zyxin regulate oral carcinoma resistance to cisplatin. -- Abstract: Ligand engagement by integrins induces receptor clustering and formation of complexes at the integrin cytoplasmic face that controls cell signaling and cytoskeletal dynamics critical for adhesion-dependent processes. This study searches for a subset of integrin effectors that coordinates both tumor cell invasion and resistance to the chemotherapeutic drug cisplatin in oral carcinomas. Candidate integrin effectors were identified in a proteomics screen of proteins recruited to clustered integrin {alpha}{beta}1, {alpha}{sub v}{beta} or {alpha}{sub 6}{beta} receptors in oral carcinomas. Proteins with diverse functions including microtubule and actin binding proteins, and factors involved in trafficking, transcription and translation were identified in oral carcinoma integrin complexes. Knockdown of effectors in the oral carcinoma HN12 cells revealed that p130Cas, Dek, Src and talin were required for invasion through Matrigel. Disruption of talin or p130Cas by RNA interference increased resistance to cisplatin, whereas targeting Dek, Src or zyxin reduced HN12 resistance to cisplatin. Analysis of the spreading of HN12 cells on collagen I and laminin I revealed that a decrease in p130Cas or talin expression inhibited spreading on both matrices. Interestingly, a reduction in zyxin expression enhanced spreading on laminin I and inhibited spreading on collagen I. Reduction of Dek, Src, talin or zyxin expression reduced HN12 proliferation by 30%. Proliferation was not affected by a reduction in p130Cas expression. We conclude that p130Cas, Src and talin function in both oral carcinoma invasion and resistance to cisplatin.

Src family kinases in chronic kidney disease.

Science.gov (United States)

Wang, Jun; Zhuang, Shougang

2017-09-01

Src family kinases (SFKs) belong to nonreceptor protein tyrosine kinases and have been implicated in the regulation of numerous cellular processes, including cell proliferation, differentiation, migration and invasion, and angiogenesis. The role and mechanisms of SFKs in tumorgenesis have been extensively investigated, and some SFK inhibitors are currently under clinical trials for tumor treatment. Recent studies have also demonstrated the importance of SFKs in regulating the development of various fibrosis-related chronic diseases (e.g., idiopathic pulmonary fibrosis, liver fibrosis, renal fibrosis, and systemic sclerosis). In this article, we summarize the roles of SFKs in various chronic kidney diseases, including glomerulonephritis, diabetic nephropathy, human immunodeficiency virus-associated nephropathy, autosomal dominant form of polycystic kidney disease, and obesity-associated kidney disease, and discuss the mechanisms involved. Copyright © 2017 the American Physiological Society.

Data provenance assurance in the cloud using blockchain

Science.gov (United States)

Shetty, Sachin; Red, Val; Kamhoua, Charles; Kwiat, Kevin; Njilla, Laurent

2017-05-01

Ever increasing adoption of cloud technology scales up the activities like creation, exchange, and alteration of cloud data objects, which create challenges to track malicious activities and security violations. Addressing this issue requires implementation of data provenance framework so that each data object in the federated cloud environment can be tracked and recorded but cannot be modified. The blockchain technology gives a promising decentralized platform to build tamper-proof systems. Its incorruptible distributed ledger/blockchain complements the need of maintaining cloud data provenance. In this paper, we present a cloud based data provenance framework using block chain which traces data record operations and generates provenance data. We anchor provenance data records into block chain transactions, which provide validation on provenance data and preserve user privacy at the same time. Once the provenance data is uploaded to the global block chain network, it is extremely challenging to tamper the provenance data. Besides, the provenance data uses hashed user identifiers prior to uploading so the blockchain nodes cannot link the operations to a particular user. The framework ensures that the privacy is preserved. We implemented the architecture on ownCloud, uploaded records to blockchain network, stored records in a provenance database and developed a prototype in form of a web service.

Berberine Reduces the Metastasis of Chondrosarcoma by Modulating the αvβ3 Integrin and the PKCδ, c-Src, and AP-1 Signaling Pathways

Directory of Open Access Journals (Sweden)

Chi-Ming Wu

2013-01-01

Full Text Available Chondrosarcoma is a primary malignant bone cancer, with a potent capacity to invade locally and cause distant metastasis, especially to the lungs. Patients diagnosed with chondrosarcoma have poor prognosis. Berberine, an active component of the Ranunculaceae and Papaveraceae families of plant, has been proven to induce tumor apoptosis and to prevent the metastasis of cancer cells. However, the effects of berberine in human chondrosarcoma are largely unknown. In this study, we found that berberine did not induce cell apoptosis in human primary chondrocytes and chondrosarcoma cells. However, at noncytotoxic concentrations, berberine reduced the migration and invasion of chondrosarcoma cancer cells. Integrins are the major adhesive molecules in mammalian cells and have been associated with the metastasis of cancer cells. We also found that incubation of chondrosarcoma cells with berberine reduced mRNA transcription for, and cell surface expression of, the αvβ3 integrin, with additional inhibitory effects on PKCδ, c-Src, and NF-κB activation. Thus, berberine may be a novel antimetastasis agent for the treatment of metastatic chondrosarcoma.

Logical provenance in data-oriented workflows?

KAUST Repository

Ikeda, R.; Das Sarma, Akash; Widom, J.

2013-01-01

for general transformations, introducing the notions of correctness, precision, and minimality. We then determine when properties such as correctness and minimality carry over from the individual transformations' provenance to the workflow provenance. We

Advanced new technologies for residue upgrading

International Nuclear Information System (INIS)

Gillis, D.

1997-01-01

Viewgraphs summarizing UOP technologies for residue are provided. The upgrading technologies include: (1) Aquaconversion, (2) the Discriminatory Destructive Distillation process (3D), and (3) the RCD uniflex process. The Aquaconversion process is based on catalytic hydrovisbreaking. It makes use of a homogeneous (liquid phase) catalyst. The hydrogen is derived from water. The advantages of this process are improved residue stability and quality at higher conversion levels, high synthetic crude yields, low operational complexity, reduced transportation costs. The 3D process is a unique carbon rejection contaminant removal process which can process whole crudes through viscous residues. FCC type equipment is used. Performance characteristics and advantages of the process were highlighted. The RCD uniflex process makes use of proven fixed bed and ebullated bed technologies in a new process configuration in which the fixed bed system is located upstream of the ebullated bed system. In this process, the catalyst volume increases exponentially with increasing processing severity. Performance characteristics, design features, benefits and development progress to date are described. 1 tab., 21 figs

Facilitating Scientific Research through Workflows and Provenance on the DataONE Cyberinfrastructure (Invited)

Science.gov (United States)

Ludaescher, B.; Cuevas-Vicenttín, V.; Missier, P.; Dey, S.; Kianmajd, P.; Wei, Y.; Koop, D.; Chirigati, F.; Altintas, I.; Belhajjame, K.; Bowers, S.

2013-12-01

Provenance data has numerous applications in science. Two key ones are 1) replication: facilitate the repeatable derivation of results and 2) discovery: enable the location of data based on processing history and derivation relationships. The following scenario illustrates a typical use of provenance data. Alice, a climate scientist, has developed a VisTrails workflow to prepare Gross Primary Productivity (GPP) data. After verifying that the workflow generates data in the desired form, she uses the ReproZip tool to create a reproducible package that will enable other scientists to re-run the workflow without having to install and configure the particular libraries she is using. In addition, she exports the provenance information of the workflow execution and customizes it through a tool such as the ProvExplorer, in order to eliminate the information she regards as superfluous. She then creates and shares a DataONE data package containing the data she prepared, the ReproZip package, the customized provenance, and additional science/system metadata. Both the customized provenance and metadata are indexed by the DataONE Cyberinfrastructure (CI) for discovery purposes. Bob, another climate scientist, is looking for a benchmark GPP data to validate the Terrestrial Biosphere Model (TBM) he has developed. Searching the DataONE repository he finds Alice's data package. He retrieves its ReproZip package, customizes it (e.g. changing the spatial resolution), and re-runs it to generate the benchmark data in the form he desires. The newly generated data is then used as input for his own model evaluation workflow. His workflow generates residual maps and a Taylor diagram that enable him to evaluate the similarity between the results of his model and the benchmark data. At this point, Bob can also make use of the tools Alice used to publish his results as another discoverable and reproducible data package. In order to support these capabilities, we propose to extend the Data

Reactivity of Athabasca residue and of its SARA fractions during residue hydroconversion

Energy Technology Data Exchange (ETDEWEB)

Verstraete, J.; Danial-Fortain, P.; Gauthier, T.; Merdrignac, I. [IFP-Lyon, Vermaison (France); Budzinski, H. [Bordeaux Univ. (France). ISM-LPTC, UMR CNRS

2009-07-01

Residue conversion processes are becoming increasingly important because of the declining market for residual fuel oil and a greater demand for middle distillates. Ebullated-bed hydroconversion is a commercially proven technology for converting heavy feedstocks with high amounts of impurities. The process enables the conversion of atmospheric or vacuum residues at temperatures up to 440 degrees C, and at liquid hourly space velocity (LHSV) conditions in the range of 0.15 to 0.5 per hour. A 540 degrees C conversion of up to 80 weight per cent can be achieved under these conditions. This paper reported on a research study conducted at IFP Lyon in which the residue hydroconversion in a large-scale ebullated bed bench unit was investigated to determine the impact of operating conditions and feed properties on yield and product qualities. Hydrogen was added to the feed in the bench units to keep a high hydrogen partial pressure and favour the catalytic hydroconversion reactions. In a typical test, the reactor was fed with 50 g of feedstock and 0.45 g of crushed equilibrium industrial NiMo catalyst, pressurized hydrogen and quickly heated at the reaction temperature. This paper also discussed the conversion of Athabasca bitumen residue in the large-scale pilot plant and also in the small scale batch reactor. The effect of operating temperature and space velocity was examined. The reactivity of the saturates, aromatics, resins and asphaltenes (SARA) fractions of the bitumen was studied separately in order to better understand the conversion mechanisms and reactivities. The Athabasca bitumen feed and SARA fractions were also analyzed in terms of standard petroleum analysis, SARA fractionation, elemental analysis, size exclusion chromatography (SEC) and 13C NMR. Hydroconversion experiments were conducted in the batch unit at different reaction temperatures and reaction times. A comparison of small-scale batch results with those obtained with the continuous large-scale bench

Gallic acid abolishes the EGFR/Src/Akt/Erk-mediated expression of matrix metalloproteinase-9 in MCF-7 breast cancer cells.

Science.gov (United States)

Chen, Ying-Jung; Lin, Ku-Nan; Jhang, Li-Mei; Huang, Chia-Hui; Lee, Yuan-Chin; Chang, Long-Sen

2016-05-25

Several studies have revealed that natural compounds are valuable resources to develop novel agents against dysregulation of the EGF/EGFR-mediated matrix metalloproteinase-9 (MMP-9) expression in cancer cells. In view of the findings that EGF/EGFR-mediated MMP-9 expression is closely related to invasion and metastasis of breast cancer. To determine the beneficial effects of gallic acid on the suppression of breast cancer metastasis, we explored the effect of gallic acid on MMP-9 expression in EGF-treated MCF-7 breast cancer cells. Treatment with EGF up-regulated MMP-9 mRNA and protein levels in MCF-7 cells. EGF treatment induced phosphorylation of EGFR and elicited Src activation, subsequently promoting Akt/NFκB (p65) and ERK/c-Jun phosphorylation in MCF-7 cells. Activation of Akt/p65 and ERK/c-Jun was responsible for the MMP-9 up-regulation in EGF-treated cells. Gallic acid repressed the EGF-induced activation of EGFR and Src; furthermore, inactivation of Akt/p65 and ERK/c-Jun was a result of the inhibitory effect of gallic acid on the EGF-induced MMP-9 up-regulation. Over-expression of constitutively active Akt and MEK1 or over-expression of constitutively active Src eradicated the inhibitory effect of gallic acid on the EGF-induced MMP-9 up-regulation. A chromosome conformation capture assay showed that EGF induced a chromosomal loop formation in the MMP-9 promoter via NFκB/p65 and AP-1/c-Jun activation. Treatment with gallic acid, EGFR inhibitor, or Src inhibitor reduced DNA looping. Taken together, our data suggest that gallic acid inhibits the activation of EGFR/Src-mediated Akt and ERK, leading to reduced levels of p65/c-Jun-mediated DNA looping and thus inhibiting MMP-9 expression in EGF-treated MCF-7 cells. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

SEIS-PROV: Practical Provenance for Seismological Data

Science.gov (United States)

Krischer, L.; Smith, J. A.; Tromp, J.

2015-12-01

It is widely recognized that reproducibility is crucial to advance science, but at the same time it is very hard to actually achieve. This results in it being recognized but also mostly ignored by a large fraction of the community. A key ingredient towards full reproducibility is to capture and describe the history of data, an issue known as provenance. We present SEIS-PROV, a practical format and data model to store provenance information for seismological data. In a seismological context, provenance can be seen as information about the processes that generated and modified a particular piece of data. For synthetic waveforms the provenance information describes which solver and settings therein were used to generate it. When looking at processed seismograms, the provenance conveys information about the different time series analysis steps that led to it. Additional uses include the description of derived data types, such as cross-correlations and adjoint sources, enabling their proper storage and exchange. SEIS-PROV is based on W3C PROV (http://www.w3.org/TR/prov-overview/), a standard for generic provenance information. It then applies an additional set of constraints to make it suitable for seismology. We present a definition of the SEIS-PROV format, a way to check if any given file is a valid SEIS-PROV document, and two sample implementations: One in SPECFEM3D GLOBE (https://geodynamics.org/cig/software/specfem3d_globe/) to store the provenance information of synthetic seismograms and another one as part of the ObsPy (http://obspy.org) framework enabling automatic tracking of provenance information during a series of analysis and transformation stages. This, along with tools to visualize and interpret provenance graphs, offers a description of data history that can be readily tracked, stored, and exchanged.

Effects of cypermethrin on the ligand-independent interaction between androgen receptor and steroid receptor coactivator-1

International Nuclear Information System (INIS)

Pan, Chen; Liu, Ya-Peng; Li, Yan-Fang; Hu, Jin-Xia; Zhang, Jin-Peng; Wang, Hong-Mei; Li, Jing; Xu, Li-Chun

2012-01-01

The pyrethroid insecticide, cypermethrin has been considered as an environmental anti-androgen by interfering with the androgen receptor (AR) transactivation. In order to clarify the effects of cypermethrin on the ligand-independent interaction between the AR and SRC-1, the mammalian two-hybrid assay has been developed in the study. The AR N-terminal domain 1–660 amino acid residues were subcloned into the plasmid pVP16 to construct the vector pVP16-ARNTD. The SRC-1 C-terminal domain 989–1240 amino acid residues were subcloned into the plasmid pM to construct the vector pM-SRC-1. The fusion vectors pVP16-ARNTD, pM-SRC-1 and the pG5CAT Reporter Vector were cotransfected into the CV-1 cells. The AR AF1 interacted with SRC-1 in the absence of exogenous ligand 5α-dihydrotestosterone (DHT). Furthermore, DHT did not enhance the interaction between AR AF-1 and SRC-1 at the concentrations from 10 −10 M to 10 −8 M. Cypermethrin inhibited the interaction between the AR AF1 and SRC-1, and the significant reduction was detected at the concentration of 10 −5 M. It is suggested that the interaction between the AR AF1 and SRC-1 is ligand-independent. Cypermethrin inhibits AR activity by disrupting the ligand-independent AR–SRC-1 interaction.

Provenance trials of larch in Siberia

Energy Technology Data Exchange (ETDEWEB)

Milyutin, L.I. [V.N. Sukachev Inst. of Forest SB RAS, Krasnoyarsk (Russian Federation)

1995-12-31

Some results of provenance trials of larch in Siberia are given. These provenance trials were established in the last thirty years by efforts of V.N. Sukaczev Inst. of Forest. Provenances and species of larch were tested in some field trials distributed over Siberia between Lat. N 52 deg and 66 deg, Long. E 88 deg and 113 deg: near Krasnoyarsk, in Republic Khakasia (an altitudes of 800 and 1200 metres), in the Lower Yenisei near Turukhansk, in the west and south regions of Krasnoyarsk territory, in the Upper Lena, near Chita. 2 refs

Archives and societal provenance Australian essays

CERN Document Server

Piggott, Michael

2012-01-01

Records and archival arrangements in Australia are globally relevant because Australia's indigenous people represent the oldest living culture in the world, and because modern Australia is an ex-colonial society now heavily multicultural in outlook. Archives and Societal Provenance explores this distinctiveness using the theoretical concept of societal provenance as propounded by Canadian archival scholars led by Dr Tom Nesmith. The book's seventeen essays blend new writing and re-workings of earlier work, comprising the fi rst text to apply a societal provenance perspective to a national sett

Provenance trials of larch in Siberia

Energy Technology Data Exchange (ETDEWEB)

Milyutin, L I [V.N. Sukachev Inst. of Forest SB RAS, Krasnoyarsk (Russian Federation)

1996-12-31

Some results of provenance trials of larch in Siberia are given. These provenance trials were established in the last thirty years by efforts of V.N. Sukaczev Inst. of Forest. Provenances and species of larch were tested in some field trials distributed over Siberia between Lat. N 52 deg and 66 deg, Long. E 88 deg and 113 deg: near Krasnoyarsk, in Republic Khakasia (an altitudes of 800 and 1200 metres), in the Lower Yenisei near Turukhansk, in the west and south regions of Krasnoyarsk territory, in the Upper Lena, near Chita. 2 refs

HIV-1 Nef interaction influences the ATP-binding site of the Src-family kinase, Hck

Directory of Open Access Journals (Sweden)

Pene-Dumitrescu Teodora

2012-03-01

Full Text Available Abstract Background Nef is an HIV-1 accessory protein essential for viral replication and AIDS progression. Nef interacts with a multitude of host cell signaling partners, including members of the Src kinase family. Nef preferentially activates Hck, a Src-family kinase (SFK strongly expressed in macrophages and other HIV target cells, by binding to its regulatory SH3 domain. Recently, we identified a series of kinase inhibitors that preferentially inhibit Hck in the presence of Nef. These compounds also block Nef-dependent HIV replication, validating the Nef-SFK signaling pathway as an antiretroviral drug target. Our findings also suggested that by binding to the Hck SH3 domain, Nef indirectly affects the conformation of the kinase active site to favor inhibitor association. Results To test this hypothesis, we engineered a "gatekeeper" mutant of Hck with enhanced sensitivity to the pyrazolopyrimidine tyrosine kinase inhibitor, NaPP1. We also modified the RT loop of the Hck SH3 domain to enhance interaction of the kinase with Nef. This modification stabilized Nef:Hck interaction in solution-based kinase assays, as a way to mimic the more stable association that likely occurs at cellular membranes. Introduction of the modified RT loop rendered Hck remarkably more sensitive to activation by Nef, and led to a significant decrease in the Km for ATP as well as enhanced inhibitor potency. Conclusions These observations suggest that stable interaction with Nef may induce Src-family kinase active site conformations amenable to selective inhibitor targeting.

Src-independent ERK signaling through the rat α3 isoform of Na/K-ATPase.

Science.gov (United States)

Madan, Namrata; Xu, Yunhui; Duan, Qiming; Banerjee, Moumita; Larre, Isabel; Pierre, Sandrine V; Xie, Zijian

2017-03-01

The Na/K-ATPase α1 polypeptide supports both ion-pumping and signaling functions. The Na/K-ATPase α3 polypeptide differs from α1 in both its primary structure and its tissue distribution. The expression of α3 seems particularly important in neurons, and recent clinical evidence supports a unique role of this isoform in normal brain function. The nature of this specific role of α3 has remained elusive, because the ubiquitous presence of α1 has hindered efforts to characterize α3-specific functions in mammalian cell systems. Using Na/K-ATPase α1 knockdown pig kidney cells (PY-17), we generated the first stable mammalian cell line expressing a ouabain-resistant form of rat Na/K-ATPase α3 in the absence of endogenous pig α1 detectable by Western blotting. In these cells, Na/K-ATPase α3 formed a functional ion-pumping enzyme and rescued the expression of Na/K-ATPase β1 and caveolin-1 to levels comparable with those observed in PY-17 cells rescued with a rat Na/K-ATPase α1 (AAC-19). The α3-containing enzymes had lower Na + affinity and lower ouabain-sensitive transport activity than their α1-containing counterparts under basal conditions, but showed a greater capacity to be activated when intracellular Na + was increased. In contrast to Na/K-ATPase α1, α3 could not regulate Src. Upon exposure to ouabain, Src activation did not occur, yet ERK was activated through Src-independent pathways involving PI3K and PKC. Hence, α3 expression confers signaling and pumping properties that are clearly distinct from that of cells expressing Na/K-ATPase α1. Copyright © 2017 the American Physiological Society.

v-src induction of the TIS10/PGS2 prostaglandin synthase gene is mediated by an ATF/CRE transcription response element.

OpenAIRE

Xie, W; Fletcher, B S; Andersen, R D; Herschman, H R

1994-01-01

We recently reported the cloning of a mitogen-inducible prostaglandin synthase gene, TIS10/PGS2. In addition to growth factors and tumor promoters, the v-src oncogene induces TIS10/PGS2 expression in 3T3 cells. Deletion analysis, using luciferase reporters, identifies a region between -80 and -40 nucleotides 5' of the TIS10/PGS2 transcription start site that mediates pp60v-src induction in 3T3 cells. This region contains the sequence CGTCACGTG, which includes overlapping ATF/CRE (CGTCA) and E...

SRC Inhibition Reduces NR2B Surface Expression and Synaptic Plasticity in the Amygdala

Science.gov (United States)

Sinai, Laleh; Duffy, Steven; Roder, John C.

2010-01-01

The Src protein tyrosine kinase plays a central role in the regulation of N-methyl-d-aspartate receptor (NMDAR) activity by regulating NMDAR subunit 2B (NR2B) surface expression. In the amygdala, NMDA-dependent synaptic plasticity resulting from convergent somatosensory and auditory inputs contributes to emotional memory; however, the role of Src…

Targeting Src family kinases inhibits bevacizumab-induced glioma cell invasion.

Directory of Open Access Journals (Sweden)

Deborah Huveldt

Full Text Available Anti-VEGF antibody therapy with bevacizumab provides significant clinical benefit in patients with recurrent glioblastoma multiforme (GBM. Unfortunately, progression on bevacizumab therapy is often associated with a diffuse disease recurrence pattern, which limits subsequent therapeutic options. Therefore, there is an urgent need to understand bevacizumab's influence on glioma biology and block it's actions towards cell invasion. To explore the mechanism(s of GBM cell invasion we have examined a panel of serially transplanted human GBM lines grown either in short-term culture, as xenografts in mouse flank, or injected orthotopically in mouse brain. Using an orthotopic xenograft model that exhibits increased invasiveness upon bevacizumab treatment, we also tested the effect of dasatinib, a broad spectrum SFK inhibitor, on bevacizumab-induced invasion.We show that 1 activation of Src family kinases (SFKs is common in GBM, 2 the relative invasiveness of 17 serially transplanted GBM xenografts correlates strongly with p120 catenin phosphorylation at Y228, a Src kinase site, and 3 SFK activation assessed immunohistochemically in orthotopic xenografts, as well as the phosphorylation of downstream substrates occurs specifically at the invasive tumor edge. Further, we show that SFK signaling is markedly elevated at the invasive tumor front upon bevacizumab administration, and that dasatinib treatment effectively blocked the increased invasion induced by bevacizumab.Our data are consistent with the hypothesis that the increased invasiveness associated with anti-VEGF therapy is due to increased SFK signaling, and support testing the combination of dasatinib with bevacizumab in the clinic.

Diallyl disulfide suppresses SRC/Ras/ERK signaling-mediated proliferation and metastasis in human breast cancer by up-regulating miR-34a.

Directory of Open Access Journals (Sweden)

Xiangsheng Xiao

Full Text Available Diallyl disulfide (DADS is one of the major volatile components of garlic oil. DADS has various biological properties, including anticancer, antiangiogenic, and antioxidant effects. However, the anticancer mechanisms of DADS in human breast cancer have not been elucidated, particularly in vivo. In this study, we demonstrated that the expression of miR-34a was up-regulated in DADS-treated MDA-MB-231 cells. miR-34a not only inhibited breast cancer growth but also enhanced the antitumor effect of DADS, both in vitro and in vivo. Furthermore, Src was identified as a target of miR-34a, with miR-34a inhibiting SRC expression and consequently triggering the suppression of the SRC/Ras/ERK pathway. These results suggest that DADS could be a promising anticancer agent for breast cancer. miR-34a may also demonstrate a potential gene therapy agent that could enhance the antitumor effects of DADS.

Lightweight Provenance Service for High-Performance Computing

Energy Technology Data Exchange (ETDEWEB)

Dai, Dong; Chen, Yong; Carns, Philip; Jenkins, John; Ross, Robert

2017-09-09

Provenance describes detailed information about the history of a piece of data, containing the relationships among elements such as users, processes, jobs, and workflows that contribute to the existence of data. Provenance is key to supporting many data management functionalities that are increasingly important in operations such as identifying data sources, parameters, or assumptions behind a given result; auditing data usage; or understanding details about how inputs are transformed into outputs. Despite its importance, however, provenance support is largely underdeveloped in highly parallel architectures and systems. One major challenge is the demanding requirements of providing provenance service in situ. The need to remain lightweight and to be always on often conflicts with the need to be transparent and offer an accurate catalog of details regarding the applications and systems. To tackle this challenge, we introduce a lightweight provenance service, called LPS, for high-performance computing (HPC) systems. LPS leverages a kernel instrument mechanism to achieve transparency and introduces representative execution and flexible granularity to capture comprehensive provenance with controllable overhead. Extensive evaluations and use cases have confirmed its efficiency and usability. We believe that LPS can be integrated into current and future HPC systems to support a variety of data management needs.

Data Provenance and Data Management in eScience

CERN Document Server

Bai, Quan; Giugni, Stephen; Williamson, Darrell; Taylor, John

2013-01-01

eScience allows scientific research to be carried out in highly distributed environments. The complex nature of the interactions in an eScience infrastructure, which often involves a range of instruments, data, models, applications, people and computational facilities, suggests there is a need for data provenance and data management (DPDM). The W3C Provenance Working Group defines the provenance of a resource as a “record that describes entities and processes involved in producing and delivering or otherwise influencing that resource”. It has been widely recognised that provenance is a critical issue to enable sharing, trust, authentication and reproducibility of eScience process.   Data Provenance and Data Management in eScience identifies the gaps between DPDM foundations and their practice within eScience domains including clinical trials, bioinformatics and radio astronomy. The book covers important aspects of fundamental research in DPDM including provenance representation and querying. It also expl...

PD-1 immunoreceptor inhibits B cell receptor-mediated signaling by recruiting src homology 2-domain-containing tyrosine phosphatase 2 to phosphotyrosine

Science.gov (United States)

Okazaki, Taku; Maeda, Akito; Nishimura, Hiroyuki; Kurosaki, Tomohiro; Honjo, Tasuku

2001-01-01

PD-1 is an immunoreceptor that belongs to the immunoglobulin (Ig) superfamily and contains two tyrosine residues in the cytoplasmic region. Studies on PD-1-deficient mice have shown that PD-1 plays critical roles in establishment and/or maintenance of peripheral tolerance, but the mode of action is totally unknown. To study the molecular mechanism for negative regulation of lymphocytes through the PD-1 receptor, we generated chimeric molecules composed of the IgG Fc receptor type IIB (FcγRIIB) extracellular region and the PD-1 cytoplasmic region and expressed them in a B lymphoma cell line, IIA1.6. Coligation of the cytoplasmic region of PD-1 with the B cell receptor (BCR) in IIA1.6 transformants inhibited BCR-mediated growth retardation, Ca2+ mobilization, and tyrosine phosphorylation of effector molecules, including Igβ, Syk, phospholipase C-γ2 (PLCγ2), and ERK1/2, whereas phosphorylation of Lyn and Dok was not affected. Mutagenesis studies indicated that these inhibitory effects do not require the N-terminal tyrosine in the immunoreceptor tyrosine-based inhibitory motif-like sequence, but do require the other tyrosine residue in the C-terminal tail. This tyrosine was phosphorylated and recruited src homology 2-domain-containing tyrosine phosphatase 2 (SHP-2) on coligation of PD-1 with BCR. These results show that PD-1 can inhibit BCR signaling by recruiting SHP-2 to its phosphotyrosine and dephosphorylating key signal transducers of BCR signaling. PMID:11698646

SRC-I demonstration plant analytical laboratory methods manual. Final technical report

Energy Technology Data Exchange (ETDEWEB)

Klusaritz, M.L.; Tewari, K.C.; Tiedge, W.F.; Skinner, R.W.; Znaimer, S.

1983-03-01

This manual is a compilation of analytical procedures required for operation of a Solvent-Refined Coal (SRC-I) demonstration or commercial plant. Each method reproduced in full includes a detailed procedure, a list of equipment and reagents, safety precautions, and, where possible, a precision statement. Procedures for the laboratory's environmental and industrial hygiene modules are not included. Required American Society for Testing and Materials (ASTM) methods are cited, and ICRC's suggested modifications to these methods for handling coal-derived products are provided.

Harvester development for new high yielding SRC crops and markets

International Nuclear Information System (INIS)

2005-12-01

Details are given of a project to develop a harvesting system that can produce fuel economically in a single pass to a required specification at a cost that is profitable for the grower while minimising the cost of the product. The project objectives listed include the development of a harvester drive chain and feeding systems to allow harvesting of the higher yielding crops now produced in the UK, determination of the most economic harvesting cycle for SRC crops, and production of fuels suitable for co-firing with coal in pulverised fuel systems or for gasification. The work programme and project conclusions are discussed

Irradiation sensibility of different provenances of Jatropha curcas L. seeds

International Nuclear Information System (INIS)

Yang Qing; Xu Congheng; Peng Daiping; Duan Zhubiao; Han Lei; Sun Qixiang; Peng Zhenhua

2007-01-01

The irradiation sensibility of 10 provenances of Jatropha curcas L. seeds to 60 Co γ-rays was studied. The results showed that the relationship between relative germination rate of the seeds and the doses of irradiation was negative correlation, and the difference of relative germination rate among different doses treatment was significant at 5% probability level or highly significant at 1% probability level. For seeds of different provenances, the correlation coefficient of linear regression was from -0.89--0.96, and the medial lethal doses (LD 50) of 10 provinces was from 127 Gy to 184 Gy. According to the LD 50, we could divided 10 provinces of J. curcas L. into sensitive provenance, transitional provenances and obtuse provenances. The provenances of Yuanjiang , Yunan (184 Gy) belonged to sensitive provenance; the provenances of Zhenfeng, Guizhou (127 Gy) and the provenances of Yuedong, Hainan (141 Gy) belonged to obtuse provenance; other 7 provenances belonged to transitional provenances. The results provided important experiment basis for germ plasma resources innovation of J. curcas L. (authors)

HBx induced AFP receptor expressed to activate PI3K/AKT signal to promote expression of Src in liver cells and hepatoma cells

International Nuclear Information System (INIS)

Zhu, Mingyue; Guo, Junli; Li, Wei; Xia, Hua; Lu, Yan; Dong, Xu; Chen, Yi; Xie, Xieju; Fu, Shigan; Li, Mengsen

2015-01-01

Hepatitis B virus (HBV)-X protein(HBx) is a transactivator of host several cellular genes including alpha-fetoprotein(AFP) and AFP receptor(AFPR) which contributes to HBV-associated tumor development. The expression of AFP/AFPR are correlated with hepatocellular carcinoma(HCC)-initial cells. But the role of AFP and AFPR in promoting occurrence of HBV-related HCC were still unclear. A total of 71 clinical patients’ liver specimens, normal human liver cells L-02 and HCC cell lines, PLC/PRF/5 were selected for analyzing the effects of HBx on expression of AFP, AFPR and Src. The expression of goal proteins were detected by Immunohistochemical stained and Western blotting; HBx-expressed vectors were constructed and transfected into L-02 cells, laser confocal microscopy was applied to observe expression and location of AFP, AFPR and Src in the normal liver cells and HCC cells, soft agar colony formation assay was used to observe colonies formed of the cells. We confirmed HBx gives preference to promote the expression of AFP and AFPR; HBx priors to up-regulate the expression of AFPR and AFP in L-02 cells and in normal liver specimens; AFPR signal been able to stimulate Src expression. The results also indicated that phosphatidylinositol 3-kinase(PI3K) inhibitors Ly294002 and GDC0941 effectively suppress AFPR mediated up-regulation expression of Src in AFPR positive HCC lines. HBx priors to drive the expression of AFP and AFPR to promote expression of Src in normal liver cells and hepatoma cells; AFP and AFPR maybe play pivotal role in HBV-related hepatocarcinogenesis; Targeting AFPR is an available therapeutic strategy of HCC. The online version of this article (doi:10.1186/s12885-015-1384-9) contains supplementary material, which is available to authorized users

Multisite tyrosine phosphorylation of the N-terminus of Mint1/X11α by Src kinase regulates the trafficking of amyloid precursor protein.

Science.gov (United States)

Dunning, Christopher J R; Black, Hannah L; Andrews, Katie L; Davenport, Elizabeth C; Conboy, Michael; Chawla, Sangeeta; Dowle, Adam A; Ashford, David; Thomas, Jerry R; Evans, Gareth J O

2016-05-01

Mint/X11 is one of the four neuronal trafficking adaptors that interact with amyloid precursor protein (APP) and are linked with its cleavage to generate β-amyloid peptide, a key player in the pathology of Alzheimer's disease. How APP switches between adaptors at different stages of the secretory pathway is poorly understood. Here, we show that tyrosine phosphorylation of Mint1 regulates the destination of APP. A canonical SH2-binding motif ((202) YEEI) was identified in the N-terminus of Mint1 that is phosphorylated on tyrosine by C-Src and recruits the active kinase for sequential phosphorylation of further tyrosines (Y191 and Y187). A single Y202F mutation in the Mint1 N-terminus inhibits C-Src binding and tyrosine phosphorylation. Previous studies observed that co-expression of wild-type Mint1 and APP causes accumulation of APP in the trans-Golgi. Unphosphorylatable Mint1 (Y202F) or pharmacological inhibition of Src reduced the accumulation of APP in the trans-Golgi of heterologous cells. A similar result was observed in cultured rat hippocampal neurons where Mint1(Y202F) permitted the trafficking of APP to more distal neurites than the wild-type protein. These data underline the importance of the tyrosine phosphorylation of Mint1 as a critical switch for determining the destination of APP. The regulation of amyloid precursor protein (APP) trafficking is poorly understood. We have discovered that the APP adapter, Mint1, is phosphorylated by C-Src kinase. Mint1 causes APP accumulation in the trans-Golgi network, whereas inhibition of Src or mutation of Mint1-Y202 permits APP recycling. The phosphorylation status of Mint1 could impact on the pathological trafficking of APP in Alzheimer's disease. © 2016 The Authors. Journal of Neurochemistry published by John Wiley & Sons Ltd on behalf of International Society for Neurochemistry.

Provenance tracking in the ViroLab Virtual Laboratory

NARCIS (Netherlands)

Baliś, B.; Bubak, M.; Wach, J.

2008-01-01

Provenance describes the process which led to the creation of a piece of data. Tracking provenance of experiment results is essential in modern environments which support conducting of in silico experiments. We present a provenance tracking approach developed as part of the virtual laboratory of the

Secure Location Provenance for Mobile Devices

Science.gov (United States)

2015-07-01

A malicious user should not be able to hide a temporary off-track movement from the claimed location provenance. • A malicious user may want to...Seltzer, “Provenance-aware storage systems,” in Proc. of USENIX ATC. USENIX Association, May 2006, pp. 43–56. [48] D. Bhagwat, L. Chiticariu, W.-C. Tan

Provenance Usage in the OceanLink Project

Science.gov (United States)

Narock, T.; Arko, R. A.; Carbotte, S. M.; Chandler, C. L.; Cheatham, M.; Fils, D.; Finin, T.; Hitzler, P.; Janowicz, K.; Jones, M.; Krisnadhi, A.; Lehnert, K. A.; Mickle, A.; Raymond, L. M.; Schildhauer, M.; Shepherd, A.; Wiebe, P. H.

2014-12-01

A wide spectrum of maturing methods and tools, collectively characterized as the Semantic Web, is helping to vastly improve thedissemination of scientific research. The OceanLink project, an NSF EarthCube Building Block, is utilizing semantic technologies tointegrate geoscience data repositories, library holdings, conference abstracts, and funded research awards. Provenance is a vital componentin meeting both the scientific and engineering requirements of OceanLink. Provenance plays a key role in justification and understanding when presenting users with results aggregated from multiple sources. In the engineering sense, provenance enables the identification of new data and the ability to determine which data sources to query. Additionally, OceanLink will leverage human and machine computation for crowdsourcing, text mining, and co-reference resolution. The results of these computations, and their associated provenance, will be folded back into the constituent systems to continually enhance precision and utility. We will touch on the various roles provenance is playing in OceanLink as well as present our use of the PROV Ontology and associated Ontology Design Patterns.

Src inhibitor herbimycin A prevents 132.7 kDa tyrosine phosphatase activity in Ramos Burkitt's lymphoma B cell line

International Nuclear Information System (INIS)

Hristov, K.; Mitev, V.; Knox, K.

2006-01-01

Reversible tyrosine phosphorylation, regulation of expression and proteolytic cleavage control tyrosine phosphatase contribution for the signalling pathways of B-cell antigen receptor (BCR), and CD40 during B cell selection. We used Ramos-BL B cell line to determine whether BCR and CD40 stimulation, or inhibition of the Src - tyrosine kinase, tyrosine phosphatase and caspase activity have an effect on the tyrosine phosphatase activities determined on in-gel phosphatase assay. The tyrosine phosphatase activities present in whole cell lysates of Ramos-BL B cells following treatment with 20 μg/ml anti-IgM, 1 μg/ml anti-CD40, 10 μM herbimycin A, 178 μM vanadate,100 μM phenylarsine oxide and 10 μM zVAD-fmk were detected with an in-gel phosphatase assay. Seven major tyrosine phosphatase activities with approximate molecular weight of 132.7, 63.9, 60.3, 54.2, 49.7, 44.6, and 39 kDa are present in whole cell lysates of Ramos-BL B cells. Treatment with Src-PTK inhibitor herbimycin A prevents 132.7 kDa tyrosine phosphatase activity. We conclude that the catalytic activity of Src-PTK in Ramos-BL B cells is critical for the presence of this 132.7 kDa tyrosine phosphatase activity. (authors)

Hydrogen Sulfide Recruits Macrophage Migration by Integrin β1-Src-FAK/Pyk2-Rac Pathway in Myocardial Infarction

Science.gov (United States)

Miao, Lei; Xin, Xiaoming; Xin, Hong; Shen, Xiaoyan; Zhu, Yi-Zhun

2016-03-01

Myocardial infarction (MI) triggers an inflammatory reaction, in which macrophages are of key importance for tissue repairing. Infiltration and/or migration of macrophages into the infarct area early after MI is critical for infarct healing, vascularization, and cardiac function. Hydrogen sulfide (H2S) has been demonstrated to possess cardioprotective effects post MI and during the progress of cardiac remodeling. However, the specific molecular and cellular mechanisms involved in macrophage recruitment by H2S remain to be identified. In this study, the NaHS (exogenous sources of H2S) treatment exerted an increased infiltration of macrophages into the infarcted myocardium at early stage of MI cardiac tissues in both wild type (WT) and cystathionine-γ-lyase-knockout (CSE-KO) mice. And NaHS accelerated the migration of macrophage cells in vitro. While, the inhibitors not only significantly diminished the migratory ability in response to NaHS, but also blocked the activation of phospho-Src, -Pyk2, -FAK397, and -FAK925. Furthermore, NaHS induced the internalization of integrin β1 on macrophage surface, but, integrin β1 silencing inhibited macrophage migration and Src signaling activation. These results indicate that H2S may have the potential as an anti-infarct of MI by governing macrophage migration, which was achieved by accelerating internalization of integrin β1 and activating downstream Src-FAK/Pyk2-Rac pathway.

The Src family kinase inhibitor dasatinib delays pain-related behaviour and conserves bone in a rat model of cancer-induced bone pain

DEFF Research Database (Denmark)

Appel, Camilla Kristine; Gallego-Pedersen, Simone; Andersen, Line

2017-01-01

Pain is a severe and debilitating complication of metastatic bone cancer. Current analgesics do not provide sufficient pain relief for all patients, creating a great need for new treatment options. The Src kinase, a non-receptor protein tyrosine kinase, is implicated in processes involved in cancer......-induced bone pain, including cancer growth, osteoclastic bone degradation and nociceptive signalling. Here we investigate the role of dasatinib, an oral Src kinase family and Bcr-Abl tyrosine kinase inhibitor, in an animal model of cancer-induced bone pain. Daily administration of dasatinib (15 mg/kg, p...

A Provenance Tracking Model for Data Updates

Directory of Open Access Journals (Sweden)

Gabriel Ciobanu

2012-08-01

Full Text Available For data-centric systems, provenance tracking is particularly important when the system is open and decentralised, such as the Web of Linked Data. In this paper, a concise but expressive calculus which models data updates is presented. The calculus is used to provide an operational semantics for a system where data and updates interact concurrently. The operational semantics of the calculus also tracks the provenance of data with respect to updates. This provides a new formal semantics extending provenance diagrams which takes into account the execution of processes in a concurrent setting. Moreover, a sound and complete model for the calculus based on ideals of series-parallel DAGs is provided. The notion of provenance introduced can be used as a subjective indicator of the quality of data in concurrent interacting systems.

LabelFlow Framework for Annotating Workflow Provenance

Directory of Open Access Journals (Sweden)

Pinar Alper

2018-02-01

Full Text Available Scientists routinely analyse and share data for others to use. Successful data (reuse relies on having metadata describing the context of analysis of data. In many disciplines the creation of contextual metadata is referred to as reporting. One method of implementing analyses is with workflows. A stand-out feature of workflows is their ability to record provenance from executions. Provenance is useful when analyses are executed with changing parameters (changing contexts and results need to be traced to respective parameters. In this paper we investigate whether provenance can be exploited to support reporting. Specifically; we outline a case-study based on a real-world workflow and set of reporting queries. We observe that provenance, as collected from workflow executions, is of limited use for reporting, as it supports queries partially. We identify that this is due to the generic nature of provenance, its lack of domain-specific contextual metadata. We observe that the required information is available in implicit form, embedded in data. We describe LabelFlow, a framework comprised of four Labelling Operators for decorating provenance with domain-specific Labels. LabelFlow can be instantiated for a domain by plugging it with domain-specific metadata extractors. We provide a tool that takes as input a workflow, and produces as output a Labelling Pipeline for that workflow, comprised of Labelling Operators. We revisit the case-study and show how Labels provide a more complete implementation of reporting queries.

Src Kinase Dependent Rapid Non-genomic Modulation of Hippocampal Spinogenesis Induced by Androgen and Estrogen

Directory of Open Access Journals (Sweden)

Mika Soma

2018-05-01

Full Text Available Dendritic spine is a small membranous protrusion from a neuron's dendrite that typically receives input from an axon terminal at the synapse. Memories are stored in synapses which consist of spines and presynapses. Rapid modulations of dendritic spines induced by hippocampal sex steroids, including dihydrotestosterone (DHT, testosterone (T, and estradiol (E2, are essential for synaptic plasticity. Molecular mechanisms underlying the rapid non-genomic modulation through synaptic receptors of androgen (AR and estrogen (ER as well as its downstream kinase signaling, however, have not been well understood. We investigated the possible involvement of Src tyrosine kinase in rapid changes of dendritic spines in response to androgen and estrogen, including DHT, T, and E2, using hippocampal slices from adult male rats. We found that the treatments with DHT (10 nM, T (10 nM, and E2 (1 nM increased the total density of spines by ~1.22 to 1.26-fold within 2 h using super resolution confocal imaging of Lucifer Yellow-injected CA1 pyramidal neurons. We examined also morphological changes of spines in order to clarify differences between three sex steroids. From spine head diameter analysis, DHT increased middle- and large-head spines, whereas T increased small- and middle-head spines, and E2 increased small-head spines. Upon application of Src tyrosine kinase inhibitor, the spine increases induced through DHT, T, and E2 treatments were completely blocked. These results imply that Src kinase is essentially involved in sex steroid-induced non-genomic modulation of the spine density and morphology. These results also suggest that rapid effects of exogenously applied androgen and estrogen can occur in steroid-depleted conditions, including “acute” hippocampal slices and the hippocampus of gonadectomized animals.

Provenance-based refresh in data-oriented workflows

KAUST Repository

Ikeda, Robert; Salihoglu, Semih; Widom, Jennifer

2011-01-01

We consider a general workflow setting in which input data sets are processed by a graph of transformations to produce output results. Our goal is to perform efficient selective refresh of elements in the output data, i.e., compute the latest values of specific output elements when the input data may have changed. We explore how data provenance can be used to enable efficient refresh. Our approach is based on capturing one-level data provenance at each transformation when the workflow is run initially. Then at refresh time provenance is used to determine (transitively) which input elements are responsible for given output elements, and the workflow is rerun only on that portion of the data needed for refresh. Our contributions are to formalize the problem setting and the problem itself, to specify properties of transformations and provenance that are required for efficient refresh, and to provide algorithms that apply to a wide class of transformations and workflows. We have built a prototype system supporting the features and algorithms presented in the paper. We report preliminary experimental results on the overhead of provenance capture, and on the crossover point between selective refresh and full workflow recomputation. © 2011 ACM.

Optimized bacterial expression and purification of the c-Src catalytic domain for solution NMR studies

International Nuclear Information System (INIS)

Piserchio, Andrea; Ghose, Ranajeet; Cowburn, David

2009-01-01

Progression of a host of human cancers is associated with elevated levels of expression and catalytic activity of the Src family of tyrosine kinases (SFKs), making them key therapeutic targets. Even with the availability of multiple crystal structures of active and inactive forms of the SFK catalytic domain (CD), a complete understanding of its catalytic regulation is unavailable. Also unavailable are atomic or near-atomic resolution information about their interactions, often weak or transient, with regulating phosphatases and downstream targets. Solution NMR, the biophysical method best suited to tackle this problem, was previously hindered by difficulties in bacterial expression and purification of sufficient quantities of soluble, properly folded protein for economically viable labeling with NMR-active isotopes. Through a choice of optimal constructs, co-expression with chaperones and optimization of the purification protocol, we have achieved the ability to bacterially produce large quantities of the isotopically-labeled CD of c-Src, the prototypical SFK, and of its activating Tyr-phosphorylated form. All constructs produce excellent spectra allowing solution NMR studies of this family in an efficient manner

Epithelial membrane protein-2 promotes endometrial tumor formation through activation of FAK and Src.

Directory of Open Access Journals (Sweden)

Maoyong Fu

Full Text Available Endometrial cancer is the most common gynecologic malignancy diagnosed among women in developed countries. One recent biomarker strongly associated with disease progression and survival is epithelial membrane protein-2 (EMP2, a tetraspan protein known to associate with and modify surface expression of certain integrin isoforms. In this study, we show using a xenograft model system that EMP2 expression is necessary for efficient endometrial tumor formation, and we have started to characterize the mechanism by which EMP2 contributes to this malignant phenotype. In endometrial cancer cells, the focal adhesion kinase (FAK/Src pathway appears to regulate migration as measured through wound healing assays. Manipulation of EMP2 levels in endometrial cancer cells regulates the phosphorylation of FAK and Src, and promotes their distribution into lipid raft domains. Notably, cells with low levels of EMP2 fail to migrate and poorly form tumors in vivo. These findings reveal the pivotal role of EMP2 in endometrial cancer carcinogenesis, and suggest that the association of elevated EMP2 levels with endometrial cancer prognosis may be causally linked to its effect on integrin-mediated signaling.

Extending eScience Provenance with User-Submitted Semantic Annotations

Science.gov (United States)

Michaelis, J.; Zednik, S.; West, P.; Fox, P. A.; McGuinness, D. L.

2010-12-01

eScience based systems generate provenance of their data products, related to such things as: data processing, data collection conditions, expert evaluation, and data product quality. Recent advances in web-based technology offer users the possibility of making annotations to both data products and steps in accompanying provenance traces, thereby expanding the utility of such provenance for others. These contributing users may have varying backgrounds, ranging from system experts to outside domain experts to citizen scientists. Furthermore, such users may wish to make varying types of annotations - ranging from documenting the purpose of a provenance step to raising concerns about the quality of data dependencies. Semantic Web technologies allow for such kinds of rich annotations to be made to provenance through the use of ontology vocabularies for (i) organizing provenance, and (ii) organizing user/annotation classifications. Furthermore, through Linked Data practices, Semantic linkages may be made from provenance steps to external data of interest. A desire for Semantically-annotated provenance has been motivated by data management issues in the Mauna Loa Solar Observatory’s (MLSO) Advanced Coronal Observing System (ACOS). In ACOS, photomoeter-based readings are taken of solar activity and subsequently processed into final data products consumable by end users. At intermediate stages of ACOS processing, factors such as evaluations by human experts and weather conditions are logged, which could impact data product quality. If such factors are linked via user-submitted annotations to provenance, it could be significantly beneficial for other users. Likewise, the background of a user could impact the credibility of their annotations. For example, an annotation made by a citizen scientist describing the purpose of a provenance step may not be as reliable as a similar annotation made by an ACOS project member. For this work, we have developed a software package that

The PBase Scientific Workflow Provenance Repository

Directory of Open Access Journals (Sweden)

Víctor Cuevas-Vicenttín

2014-10-01

Full Text Available Scientific workflows and their supporting systems are becoming increasingly popular for compute-intensive and data-intensive scientific experiments. The advantages scientific workflows offer include rapid and easy workflow design, software and data reuse, scalable execution, sharing and collaboration, and other advantages that altogether facilitate “reproducible science”. In this context, provenance – information about the origin, context, derivation, ownership, or history of some artifact – plays a key role, since scientists are interested in examining and auditing the results of scientific experiments. However, in order to perform such analyses on scientific results as part of extended research collaborations, an adequate environment and tools are required. Concretely, the need arises for a repository that will facilitate the sharing of scientific workflows and their associated execution traces in an interoperable manner, also enabling querying and visualization. Furthermore, such functionality should be supported while taking performance and scalability into account. With this purpose in mind, we introduce PBase: a scientific workflow provenance repository implementing the ProvONE proposed standard, which extends the emerging W3C PROV standard for provenance data with workflow specific concepts. PBase is built on the Neo4j graph database, thus offering capabilities such as declarative and efficient querying. Our experiences demonstrate the power gained by supporting various types of queries for provenance data. In addition, PBase is equipped with a user friendly interface tailored for the visualization of scientific workflow provenance data, making the specification of queries and the interpretation of their results easier and more effective.

ROS mediates interferon gamma induced phosphorylation of Src, through the Raf/ERK pathway, in MCF-7 human breast cancer cell line.

Science.gov (United States)

Zibara, Kazem; Zeidan, Asad; Bjeije, Hassan; Kassem, Nouhad; Badran, Bassam; El-Zein, Nabil

2017-03-01

Interferon gamma (IFN-ɣ) is a pleiotropic cytokine which plays dual contrasting roles in cancer. Although IFN-ɣ has been clinically used to treat various malignancies, it was recently shown to have protumorigenic activities. Reactive oxygen species (ROS) are overproduced in cancer cells, mainly due to NADPH oxidase activity, which results into several changes in signaling pathways. In this study, we examined IFN-ɣ effect on the phosphorylation levels of key signaling proteins, through ROS production, in the human breast cancer cell line MCF-7. After treatment by IFN-ɣ, results showed a significant increase in the phosphorylation of STAT1, Src, raf, AKT, ERK1/2 and p38 signaling molecules, in a time specific manner. Src and Raf were found to be involved in early stages of IFN-ɣ signaling since their phosphorylation increased very rapidly. Selective inhibition of Src-family kinases resulted in an immediate significant decrease in the phosphorylation status of Raf and ERK1/2, but not p38 and AKT. On the other hand, IFN-ɣ resulted in ROS generation, through H 2 O 2 production, whereas pre-treatment with the ROS inhibitor NAC caused ROS inhibition and a significant decrease in the phosphorylation levels of AKT, ERK1/2, p38 and STAT1. Moreover, pretreatment with a selective NOX1 inhibitor resulted in a significant decrease of AKT phosphorylation. Finally, no direct relationship was found between ROS production and calcium mobilization. In summary, IFN-ɣ signaling in MCF-7 cell line is ROS-dependent and follows the Src/Raf/ERK pathway whereas its signaling through the AKT pathway is highly dependent on NOX1.

PAV ontology: provenance, authoring and versioning.

Science.gov (United States)

Ciccarese, Paolo; Soiland-Reyes, Stian; Belhajjame, Khalid; Gray, Alasdair Jg; Goble, Carole; Clark, Tim

2013-11-22

Provenance is a critical ingredient for establishing trust of published scientific content. This is true whether we are considering a data set, a computational workflow, a peer-reviewed publication or a simple scientific claim with supportive evidence. Existing vocabularies such as Dublin Core Terms (DC Terms) and the W3C Provenance Ontology (PROV-O) are domain-independent and general-purpose and they allow and encourage for extensions to cover more specific needs. In particular, to track authoring and versioning information of web resources, PROV-O provides a basic methodology but not any specific classes and properties for identifying or distinguishing between the various roles assumed by agents manipulating digital artifacts, such as author, contributor and curator. We present the Provenance, Authoring and Versioning ontology (PAV, namespace http://purl.org/pav/): a lightweight ontology for capturing "just enough" descriptions essential for tracking the provenance, authoring and versioning of web resources. We argue that such descriptions are essential for digital scientific content. PAV distinguishes between contributors, authors and curators of content and creators of representations in addition to the provenance of originating resources that have been accessed, transformed and consumed. We explore five projects (and communities) that have adopted PAV illustrating their usage through concrete examples. Moreover, we present mappings that show how PAV extends the W3C PROV-O ontology to support broader interoperability. The initial design of the PAV ontology was driven by requirements from the AlzSWAN project with further requirements incorporated later from other projects detailed in this paper. The authors strived to keep PAV lightweight and compact by including only those terms that have demonstrated to be pragmatically useful in existing applications, and by recommending terms from existing ontologies when plausible. We analyze and compare PAV with related

Evidence for in vivo phosphorylation of the Grb2 SH2-domain binding site on focal adhesion kinase by Src-family protein-tyrosine kinases.

OpenAIRE

Schlaepfer, D D; Hunter, T

1996-01-01

Focal adhesion kinase (FAK) is a nonreceptor protein-tyrosine kinase (PTK) that associates with integrin receptors and participates in extracellular matrix-mediated signal transduction events. We showed previously that the c-Src nonreceptor PTK and the Grb2 SH2/SH3 adaptor protein bound directly to FAK after fibronectin stimulation (D. D. Schlaepfer, S.K. Hanks, T. Hunter, and P. van der Geer, Nature [London] 372:786-791, 1994). Here, we present evidence that c-Src association with FAK is req...

Big Data Provenance: Challenges, State of the Art and Opportunities.

Science.gov (United States)

Wang, Jianwu; Crawl, Daniel; Purawat, Shweta; Nguyen, Mai; Altintas, Ilkay

2015-01-01

Ability to track provenance is a key feature of scientific workflows to support data lineage and reproducibility. The challenges that are introduced by the volume, variety and velocity of Big Data, also pose related challenges for provenance and quality of Big Data, defined as veracity. The increasing size and variety of distributed Big Data provenance information bring new technical challenges and opportunities throughout the provenance lifecycle including recording, querying, sharing and utilization. This paper discusses the challenges and opportunities of Big Data provenance related to the veracity of the datasets themselves and the provenance of the analytical processes that analyze these datasets. It also explains our current efforts towards tracking and utilizing Big Data provenance using workflows as a programming model to analyze Big Data.

The Symbiotic Relationship between Scientific Workflow and Provenance (Invited)

Science.gov (United States)

Stephan, E.

2010-12-01

The purpose of this presentation is to describe the symbiotic nature of scientific workflows and provenance. We will also discuss the current trends and real world challenges facing these two distinct research areas. Although motivated differently, the needs of the international science communities are the glue that binds this relationship together. Understanding and articulating the science drivers to these communities is paramount as these technologies evolve and mature. Originally conceived for managing business processes, workflows are now becoming invaluable assets in both computational and experimental sciences. These reconfigurable, automated systems provide essential technology to perform complex analyses by coupling together geographically distributed disparate data sources and applications. As a result, workflows are capable of higher throughput in a shorter amount of time than performing the steps manually. Today many different workflow products exist; these could include Kepler and Taverna or similar products like MeDICI, developed at PNNL, that are standardized on the Business Process Execution Language (BPEL). Provenance, originating from the French term Provenir “to come from”, is used to describe the curation process of artwork as art is passed from owner to owner. The concept of provenance was adopted by digital libraries as a means to track the lineage of documents while standards such as the DublinCore began to emerge. In recent years the systems science community has increasingly expressed the need to expand the concept of provenance to formally articulate the history of scientific data. Communities such as the International Provenance and Annotation Workshop (IPAW) have formalized a provenance data model. The Open Provenance Model, and the W3C is hosting a provenance incubator group featuring the Proof Markup Language. Although both workflows and provenance have risen from different communities and operate independently, their mutual

Qualitative screening method for pesticide residues detection in fruits and vegetables

Directory of Open Access Journals (Sweden)

Iván Mauricio Huérfano Barco

2018-01-01

Full Text Available Because of the importance of developing methodologies that allow agricultural residues analysis, a rapid screening qualitative method for the determination of pesticides residues in fruits and vegetables was validated. The methodology was based on the European QuEChERS extraction method with an additional cleaning step by gel permeation chromatography (GPC, which helped to reduce the number of matrix components in the final extract. The analysis was carried out by gas chromatography coupled to mass spectrometry with a single quadrupole analyzer. The methodology was appropriate for the qualitative analysis of 31 pesticides at their respective maximum residue limits. Consistent results were obtained with respect to a quantitative routine methodology in the analysis of real samples, hence the methodology was proven to be a good alternative for the fast analysis of these contaminants in fruits and vegetables.

Type I collagen synergistically enhances PDGF-induced smooth muscle cell proliferation through pp60src-dependent crosstalk between the α2β1 integrin and PDGFβ receptor

International Nuclear Information System (INIS)

Hollenbeck, Scott T.; Itoh, Hiroyuki; Louie, Otway; Faries, Peter L.; Liu Bo; Kent, K. Craig

2004-01-01

Smooth muscle cells (SMCs) are exposed to both platelet-derived growth factor (PDGF) and type I collagen (CNI) at the time of arterial injury. In these studies we explore the individual and combined effects of these agonists on human saphenous vein SMC proliferation. PDGF-BB produced a 5.5-fold increase in SMC DNA synthesis whereas CNI stimulated DNA synthesis to a much lesser extent (1.6-fold increase). Alternatively, we observed an 8.3-fold increase in DNA synthesis when SMCs were co-incubated with CNI and PDGF-BB. Furthermore, stimulation of SMCs with PDGF-BB produced a significant increase in ERK-2 activity whereas CNI alone had no effect. Co-incubation of SMCs with PDGF-BB and CNI resulted in ERK-2 activity that was markedly greater than that produced by PDGF-BB alone. In a similar fashion, PDGF-BB induced phosphorylation of the PDGF receptor β (PDGFRβ) and CNI did not, whereas concurrent agonist stimulation produced a synergistic increase in receptor activity. Blocking antibodies to the α2 and β1 subunits eliminated this synergistic interaction, implicating the α2β1 integrin as the mediator of this effect. Immunoprecipitation of the α2β1 integrin in unstimulated SMCs followed by immunoblotting for the PDGFRβ as well as Src family members, pp60 src , Fyn, Lyn, and Yes demonstrated coassociation of α2β1 and the PDGFRβ as well as pp60 src . Incubation of cells with CNI and/or PDGF-BB did not change the degree of association. Finally, inhibition of Src activity with SU6656 eliminated the synergistic effect of CNI on PDGF-induced PDGFRβ phosphorylation suggesting an important role for pp60 src in the observed receptor crosstalk. Together, these data demonstrate that CNI synergistically enhances PDGF-induced SMC proliferation through Src-dependent crosstalk between the α2β1 integrin and the PDGFRβ

Model-based Abstraction of Data Provenance

DEFF Research Database (Denmark)

Probst, Christian W.; Hansen, René Rydhof

2014-01-01

to bigger models, and the analyses adapt accordingly. Our approach extends provenance both with the origin of data, the actors and processes involved in the handling of data, and policies applied while doing so. The model and corresponding analyses are based on a formal model of spatial and organisational......Identifying provenance of data provides insights to the origin of data and intermediate results, and has recently gained increased interest due to data-centric applications. In this work we extend a data-centric system view with actors handling the data and policies restricting actions....... This extension is based on provenance analysis performed on system models. System models have been introduced to model and analyse spatial and organisational aspects of organisations, to identify, e.g., potential insider threats. Both the models and analyses are naturally modular; models can be combined...

Provenance-Based Debugging and Drill-Down in Data-Oriented Workflows

KAUST Repository

Ikeda, Robert

2012-04-01

Panda (for Provenance and Data) is a system that supports the creation and execution of data-oriented workflows, with automatic provenance generation and built-in provenance tracing operations. Workflows in Panda are arbitrary a cyclic graphs containing both relational (SQL) processing nodes and opaque processing nodes programmed in Python. For both types of nodes, Panda generates logical provenance - provenance information stored at the processing-node level - and uses the generated provenance to support record-level backward tracing and forward tracing operations. In our demonstration we use Panda to integrate, process, and analyze actual education data from multiple sources. We specifically demonstrate how Panda\\'s provenance generation and tracing capabilities can be very useful for workflow debugging, and for drilling down on specific results of interest. © 2012 IEEE.

Syk/Src Pathway-Targeted Inhibition of Skin Inflammatory Responses by Carnosic Acid

Directory of Open Access Journals (Sweden)

Jueun Oh

2012-01-01

Full Text Available Carnosic acid (CA is a diterpene compound exhibiting antioxidative, anticancer, anti-angiogenic, anti-inflammatory, anti-metabolic disorder, and hepatoprotective and neuroprotective activities. In this study, the effect of CA on various skin inflammatory responses and its inhibitory mechanism were examined. CA strongly suppressed the production of IL-6, IL-8, and MCP-1 from keratinocyte HaCaT cells stimulated with sodium lauryl sulfate (SLS and retinoic acid (RA. In addition, CA blocked the release of nitric oxide (NO, tumor necrosis factor (TNF-α, and prostaglandin E2 (PGE2 from RAW264.7 cells activated by the toll-like receptor (TLR-2 ligands, Gram-positive bacterium-derived peptidoglycan (PGN and pam3CSK, and the TLR4 ligand, Gram-negative bacterium-derived lipopolysaccharide (LPS. CA arrested the growth of dermatitis-inducing Gram-positive and Gram-negative microorganisms such Propionibacterium acnes, Pseudomonas aeruginosa, and Staphylococcus aureus. CA also blocked the nuclear translocation of nuclear factor (NF-κB and its upstream signaling including Syk/Src, phosphoinositide 3-kinase (PI3K, Akt, inhibitor of κBα (IκBα kinase (IKK, and IκBα for NF-κB activation. Kinase assays revealed that Syk could be direct enzymatic target of CA in its anti-inflammatory action. Therefore, our data strongly suggest the potential of CA as an anti-inflammatory drug against skin inflammatory responses with Src/NF-κB inhibitory properties.

Power generation with technology innovation of residual biomass utilization; Geracao de energia com inovacao tecnologica de aproveitamento de biomassa residual

Energy Technology Data Exchange (ETDEWEB)

Moura, Johnson Pontes de; Selvam, P.V. Pannir [Universidade Federal do Rio Grande do Norte (UFRN), Natal, RN (Brazil)

2006-07-01

In the present work, the introduction of alternative energy of biogas in agricultural communities for the sustainable development was studied through exploitation of residual biomass and also getting as by-product the biological fertilizer. A fast composting of the domestic residue with the organic was made possible where part of this residue after processing was taken together with effluent to the biodigester. The bibliographical research on the processes of generation of biogas, about composting and the equipment for processing had been carried through. The projects engineering with the use of computational tools had been developed with the software Super Pro 4,9 Design and ORC GPEC 2004 by our research group. Five case studies had been elaborated, where different scenes related with our innovation, that uses of the residue for the composting together with domestic effluent for digestion. Several economic parameters were obtained and our work proved the viability about the use of biogas for drying of the fruits banana. A economic feasibility study was carried where it was proven that the project with the innovation of the use of residues from the fruits possess more advantages than the conventional system of drying using electric energy. Considering the viability of this process and the use solar energy, it is intended to apply this technology in rural agricultural communities providing them an energy source of low cost in substitution of the conventional energy. (author)

Provenance an introduction to PROV

CERN Document Server

Moreau, Luc

2013-01-01

The World Wide Web is now deeply intertwined with our lives, and has become a catalyst for a data deluge, making vast amounts of data available online, at a click of a button. With Web 2.0, users are no longer passive consumers, but active publishers and curators of data. Hence, from science to food manufacturing, from data journalism to personal well-being, from social media to art, there is a strong interest in provenance, a description of what influenced an artifact, a data set, a document, a blog, or any resource on the Web and beyond. Provenance is a crucial piece of information that can

Particle diffusion from resonance islands in Aladdin at SRC

International Nuclear Information System (INIS)

Liu, J.; Crosbie, E.; Teng, L.; Bridges, J.; Ciarlette, D.; Kustom, R.; Voss, D.; Mills, F.; Borland, M.; Symon, K.

1993-01-01

The dynamics of the beam in the resonance islands was studied on the electron storage ring Aladdin at the Synchrotron Radiation Center (SRC). The authors especially studied the horizontal third- and fourth-integral resonances driven by sextupole fields in the first and second order. A fast kicker was fired to kick the beam into one of the outboard stable islands. The beam took on a quasi-Gaussian distribution and slowly diffused out of the island. The diffusion rate and its dependence on the strengths of the driving sextupoles and the chromaticity sextupoles were measured by tracing the resonance peak of the betatron oscillation on the spectrum analyzer. Beam positions were also recorded through the data acquisition device which was locked by a pulse-delay circuitry. Interesting results are shown and compared with numerical calculations

Particle diffusion from resonance islands in Aladdin at SRC

International Nuclear Information System (INIS)

Liu, J.; Crosbie, E.; Teng, L.; Bridges, J.; Ciarlette, D.; Kustom, R.; Voss, D.; Mills, F.; Borland, M.; Symon, K.

1993-01-01

The dynamics of the beam in the resonance islands was studied on the electron storage ring Aladdin at the Synchrotron Radiation Center (SRC). The authors especially studied the horizontal third- and fourth-integral resonances driven by sextupole fields in the first and second order. A fast kicker was fired to kick the beam into one of the outboard stable islands. The beam took on a quasi-Gaussian distribution and slowly diffused out of the island. The diffusion rate and its dependence on the strengths of the driving sextupoles and the chromaticity sextupoles were measured by tracing the resonance peak of the betatron oscillation on the spectrum analyzer. Beam positions were also recorded through the data acquisition device which was clocked by a pulse-delay circuitry. Interesting results are shown and compared with numerical calculations

Activation of AMPA receptor promotes TNF-α release via the ROS-cSrc-NFκB signaling cascade in RAW264.7 macrophages

Energy Technology Data Exchange (ETDEWEB)

Cheng, Xiu-Li [Department of Physiology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, School of Basic Medicine, Peking Union Medical College, Beijing (China); Ding, Fan [Office of Scientific R& D, Tsinghua University, Beijing (China); Li, Hui; Tan, Xiao-Qiu [Department of Physiology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, School of Basic Medicine, Peking Union Medical College, Beijing (China); Liu, Xiao [Department of Pathophysiology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, School of Basic Medicine, Peking Union Medical College, Beijing (China); Cao, Ji-Min, E-mail: caojimin@126.com [Department of Physiology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, School of Basic Medicine, Peking Union Medical College, Beijing (China); Gao, Xue, E-mail: longlongnose@163.com [Department of Pathophysiology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, School of Basic Medicine, Peking Union Medical College, Beijing (China)

2015-05-29

The relationship between glutamate signaling and inflammation has not been well defined. This study aimed to investigate the role of AMPA receptor (AMPAR) in the expression and release of tumor necrosis factor-alpha (TNF-α) from macrophages and the underlying mechanisms. A series of approaches, including confocal microscopy, immunofluorescency, flow cytometry, ELISA and Western blotting, were used to estimate the expression of AMPAR and downstream signaling molecules, TNF-α release and reactive oxygen species (ROS) generation in the macrophage-like RAW264.7 cells. The results demonstrated that AMPAR was expressed in RAW264.7 cells. AMPA significantly enhanced TNF-α release from RAW264.7 cells, and this effect was abolished by CNQX (AMPAR antagonist). AMPA also induced elevation of ROS production, phosphorylation of c-Src and activation of nuclear factor (NF)-κB in RAW264.7 cells. Blocking c-Src by PP2, scavenging ROS by glutathione (GSH) or inhibiting NF-κB activation by pyrrolidine dithiocarbamate (PDTC) decreased TNF-α production from RAW264.7 cells. We concluded that AMPA promotes TNF-α release in RAW264.7 macrophages likely through the following signaling cascade: AMPAR activation → ROS generation → c-Src phosphorylation → NF-κB activation → TNF-α elevation. The study suggests that AMPAR may participate in macrophage activation and inflammation. - Highlights: • AMPAR is expressed in RAW264.7 macrophages and is upregulated by AMPA stimulation. • Activation of AMPAR stimulates TNF-α release in macrophages through the ROS-cSrc-NFκB signaling cascade. • Macrophage AMPAR signaling may play an important role in inflammation.

Solvent refined coal (SRC) process. Quarterly technical progress report, January 1980-March 1980. [In process streams

Energy Technology Data Exchange (ETDEWEB)

1981-01-01

This report summarizes the progress of the Solvent Refined Coal (SRC) project at the SRC Pilot Plant in Fort Lewis, Wahsington, and the Process Development Unit (P-99) in Harmarville, Pennsylvania. After the remaining runs of the slurry preheater survey test program were completed January 14, the Fort Lewis Pilot Plant was shut down to inspect Slurry Preheater B and to insulate the coil for future testing at higher rates of heat flux. Radiographic inspection of the coil showed that the welds at the pressure taps and the immersion thermowells did not meet design specifications. Slurry Preheater A was used during the first 12 days of February while weld repairs and modifications to Slurry Preheater B were completed. Two attempts to complete a material balance run on Powhatan No. 6 Mine coal were attempted but neither was successful. Slurry Preheater B was in service the remainder of the quarter. The start of a series of runs at higher heat flux was delayed because of plugging in both the slurry and the hydrogen flow metering systems. Three baseline runs and three slurry runs of the high heat flux program were completed before the plant was shut down March 12 for repair of the Inert Gas Unit. Attempts to complete a fourth slurry run at high heat flux were unsuccessful because of problems with the coal feed handling and the vortex mix systems. Process Development Unit (P-99) completed three of the four runs designed to study the effect of dissolver L/D ratio. The fourth was under way at the end of the period. SRC yield correlations have been developed that include coal properties as independent variables. A preliminary ranking of coals according to their reactivity in PDU P-99 has been made. Techniques for studying coking phenomenona are now in place.

Role of pp60(c-src) and p(44/42) MAPK in ANG II-induced contraction of rat tonic gastrointestinal smooth muscles.

Science.gov (United States)

Puri, Rajinder N; Fan, Ya-Ping; Rattan, Satish

2002-08-01

We examined the role of mitogen-activated protein kinase (p(44/42) MAPK) in ANG II-induced contraction of lower esophageal sphincter (LES) and internal anal sphincter (IAS) smooth muscles. Studies were performed in the isolated smooth muscles and cells (SMC). ANG II-induced changes in the levels of phosphorylation of different signal transduction and effector proteins were determined before and after selective inhibitors. ANG II-induced contraction of the rat LES and IAS SMC was inhibited by genistein, PD-98059 [a specific inhibitor of MAPK kinases (MEK 1/2)], herbimycin A (a pp60(c-src) inhibitor), and antibodies to pp60(c-src) and p(120) ras GTPase-activating protein (p(120) rasGAP). ANG II-induced contraction of the tonic smooth muscles was accompanied by an increase in tyrosine phosphorylation of p(120) rasGAP. These were attenuated by genistein but not by PD-98059. ANG II-induced increase in phosphorylations of p(44/42) MAPKs and caldesmon was attenuated by both genistein and PD-98059. We conclude that pp60(c-src) and p(44/42) MAPKs play an important role in ANG II-induced contraction of LES and IAS smooth muscles.

The impact of seeds provenance and nursery provenance method on Austrian pine (Pinus nigra Arn. seedlings quality

Directory of Open Access Journals (Sweden)

Vladan Ivetić

2013-12-01

Full Text Available The influence of seed provenances and seedling production methods on quality of one and two years old seedlings of Austrian pine were investigated. Seeds from three provenances of Austrian pine (Goc, Studenica and Sargan were used for seedlings production, combined with three production methods: (i the modified seedbeds (bare-root, (ii the container type Plantagrah II and (iii the container type Gocko. Provenance, as well as the combined influence of provenance and production method had minimal influence on the variability of one and two years old Austrian pine seedlings. Nevertheless, the production method had the highest influence. The production system, besides its importance in nursery, will have a high influence on seedlings growth during the first year after planting. Considering the results of this study (e.g. the highest values of the diameter, number of lateral roots, shoot and root dry weight, and quality index and the lowest value of SQ and satisfactory value of S:R, we can conclude that the seedlings produced in container type Gocko led to the highest seedlings quality, recommended especially for afforestation on hard sites.     

Provenance research: investigation of genetic diversity associated with geography

Science.gov (United States)

Robert Z. Callaham

1963-01-01

Provenance in forestry refers to the population of trees growing at n particular place of origin. Provenance research defines the genetic and environmental components of phenotypic variation associated with geographic source. Information on provenance is important in assuring sources of seed to give well-adapted, productive trees and in directing breeding of...

Provenance Store Evaluation

Energy Technology Data Exchange (ETDEWEB)

Paulson, Patrick R.; Gibson, Tara D.; Schuchardt, Karen L.; Stephan, Eric G.

2008-03-01

Requirements for the provenance store and access API are developed. Existing RDF stores and APIs are evaluated against the requirements and performance benchmarks. The team’s conclusion is to use MySQL as a database backend, with a possible move to Oracle in the near-term future. Both Jena and Sesame’s APIs will be supported, but new code will use the Jena API

Utilizing Provenance in Reusable Research Objects

Directory of Open Access Journals (Sweden)

Zhihao Yuan

2018-03-01

Full Text Available Science is conducted collaboratively, often requiring the sharing of knowledge about computational experiments. When experiments include only datasets, they can be shared using Uniform Resource Identifiers (URIs or Digital Object Identifiers (DOIs. An experiment, however, seldom includes only datasets, but more often includes software, its past execution, provenance, and associated documentation. The Research Object has recently emerged as a comprehensive and systematic method for aggregation and identification of diverse elements of computational experiments. While a necessary method, mere aggregation is not sufficient for the sharing of computational experiments. Other users must be able to easily recompute on these shared research objects. Computational provenance is often the key to enable such reuse. In this paper, we show how reusable research objects can utilize provenance to correctly repeat a previous reference execution, to construct a subset of a research object for partial reuse, and to reuse existing contents of a research object for modified reuse. We describe two methods to summarize provenance that aid in understanding the contents and past executions of a research object. The first method obtains a process-view by collapsing low-level system information, and the second method obtains a summary graph by grouping related nodes and edges with the goal to obtain a graph view similar to application workflow. Through detailed experiments, we show the efficacy and efficiency of our algorithms.

Area 3, SRC-II coal slurry preheater studies report for the technical data analysis program

Energy Technology Data Exchange (ETDEWEB)

1984-08-01

This report reviews the raw data gathered from the Preheater B test runs at Ft. Lewis, and also the Preheater B results presented in the Solvent Refined Coal (SRC) Process Final Report, Volumes 1 and 2 of Slurry Preheater Design, SRC-II Process and the Ft. Lewis Slurry Preheater Data Analysis, 1 1/2 Inch Coil by Gulf Science and Technology Corporation of Pittsburgh, Pennsylvania. attempts were made to correlate several variables not previously considered with slurry viscosity and thermal conductivity. Only partial success was realized. However, in the process of attempting to correlate these variables an understanding of why some variables could not be correlated was achieved. An attempt was also made, using multiple linear regression, to correlate coal slurry viscosity and thermal conductivity with several independent variables among which were temperature, coal concentration, total solids, coal type, slurry residence time, shear rate, and unit size. The final correlations included some, but not all, of these independent variables. This report is not a stand alone document and should be considered a supplement to work already done. It should be read in conjunction with the reports referenced above.

Combustion quality of poplar and willow clones grown as SRC at four sites in Brandenburg, Germany

DEFF Research Database (Denmark)

Liu, Na; Ugilt Larsen, Søren; Jørgensen, Uffe

2017-01-01

The fuel quality was assessed for nine poplar clones (AF2, Androscoggin, Max1, Max3, Max4, Monviso, Muhle-Larsen, NE42, Weser6) and one willow clone (Inger) grown as short rotation coppice (SRC) on four sites in the Brandenburg area in Germany. Fuel quality was analysed in 3-year old shoots in te...

Systems for harvesting and handling cotton plant residue

Energy Technology Data Exchange (ETDEWEB)

Coates, W. [Univ. of Arizona, Tucson, AZ (United States)

1993-12-31

In the warmer regions of the United States, cotton plant residue must be buried to prevent it from serving as an overwintering site for insect pests such as the pink bollworm. Most of the field operations used to bury the residue are high energy consumers and tend to degrade soil structure, thereby increasing the potential for erosion. The residue is of little value as a soil amendment and consequently is considered a negative value biomass. A commercial system to harvest cotton plant residue would be of both economic and environmental benefit to cotton producers. Research has been underway at the University of Arizona since the spring of 1991 to develop a commercially viable system for harvesting cotton plant residue. Equipment durability, degree of densification, energy required, cleanliness of the harvested material, and ease of product handling and transport are some of the performance variables which have been measured. Two systems have proven superior. In both, the plants are pulled from the ground using an implement developed specifically for the purpose. In one system, the stalks are baled using a large round baler, while in the other the stalks are chopped with a forage harvester, and then made into packages using a cotton module maker. Field capacities, energy requirements, package density and durability, and ease of handling with commercially available equipment have been measured for both systems. Selection of an optimum system for a specific operation depends upon end use of the product, and upon equipment availability.

Bridging the provenance gap: opportunities and challenges tracking in and ex silico provenance in sUAS workflows

Science.gov (United States)

Thomer, A.

2017-12-01

Data provenance - the record of the varied processes that went into the creation of a dataset, as well as the relationships between resulting data objects - is necessary to support the reusability, reproducibility and reliability of earth science data. In sUAS-based research, capturing provenance can be particularly challenging because of the breadth and distributed nature of the many platforms used to collect, process and analyze data. In any given project, multiple drones, controllers, computers, software systems, sensors, cameras, imaging processing algorithms and data processing workflows are used over sometimes long periods of time. These platforms and processing result in dozens - if not hundreds - of data products in varying stages of readiness-for-analysis and sharing. Provenance tracking mechanisms are needed to make the relationships between these many data products explicit, and therefore more reusable and shareable. In this talk, I discuss opportunities and challenges in tracking provenance in sUAS-based research, and identify gaps in current workflow-capture technologies. I draw on prior work conducted as part of the IMLS-funded Site-Based Data Curation project in which we developed methods of documenting in and ex silico (that is, computational and non-computation) workflows, and demonstrate this approaches applicability to research with sUASes. I conclude with a discussion of ontologies and other semantic technologies that have potential application in sUAS research.

Provenance-Based Debugging and Drill-Down in Data-Oriented Workflows

KAUST Repository

Ikeda, Robert; Cho, Junsang; Fang, Charlie; Salihoglu, Semih; Torikai, Satoshi; Widom, Jennifer

2012-01-01

Panda (for Provenance and Data) is a system that supports the creation and execution of data-oriented workflows, with automatic provenance generation and built-in provenance tracing operations. Workflows in Panda are arbitrary a cyclic graphs

HCV NS5A protein containing potential ligands for both Src homology 2 and 3 domains enhances autophosphorylation of Src family kinase Fyn in B cells.

Science.gov (United States)

Nakashima, Kenji; Takeuchi, Kenji; Chihara, Kazuyasu; Horiguchi, Tomoko; Sun, Xuedong; Deng, Lin; Shoji, Ikuo; Hotta, Hak; Sada, Kiyonao

2012-01-01

Hepatitis C virus (HCV) infects B lymphocytes and induces mixed cryoglobulinemia and B cell non-Hodgkin's lymphoma. The molecular mechanism for the pathogenesis of HCV infection-mediated B cell disorders remains obscure. To identify the possible role for HCV nonstructural 5A (NS5A) protein in B cells, we generated the stable B cell lines expressing Myc-His tagged NS5A. Immunoprecipitation study in the presence or absence of pervanadate (PV) implied that NS5A was tyrosine phosphorylated by pervanadate (PV) treatment of the cells. Therefore we examined pull-down assay by using glutathione S-transferase (GST)-fusion proteins of various Src homology 2 (SH2) domains, which associates with phosphotyrosine within a specific amino acid sequence. The results showed that NS5A specifically bound to SH2 domain of Fyn from PV-treated B cells in addition to Src homology 3 (SH3) domain. Substitution of Arg(176) to Lys in the SH2 domain of Fyn abrogated this interaction. Deletion mutational analysis demonstrated that N-terminal region of NS5A was not required for the interaction with the SH2 domain of Fyn. Tyr(334) was identified as a tyrosine phosphorylation site in NS5A. Far-western analysis revealed that SH2 domain of Fyn directly bound to NS5A. Fyn and NS5A were colocalized in the lipid raft. These results suggest that NS5A directly binds to the SH2 domain of Fyn in a tyrosine phosphorylation-dependent manner. Lastly, we showed that the expression of NS5A in B cells increased phosphorylation of activation loop tyrosine in the kinase domain of Fyn. NS5A containing ligand for both SH2 and SH3 domains enhances an aberrant autophosphorylation and kinase activity of Fyn in B cells.

HCV NS5A protein containing potential ligands for both Src homology 2 and 3 domains enhances autophosphorylation of Src family kinase Fyn in B cells.

Directory of Open Access Journals (Sweden)

Kenji Nakashima

Full Text Available Hepatitis C virus (HCV infects B lymphocytes and induces mixed cryoglobulinemia and B cell non-Hodgkin's lymphoma. The molecular mechanism for the pathogenesis of HCV infection-mediated B cell disorders remains obscure. To identify the possible role for HCV nonstructural 5A (NS5A protein in B cells, we generated the stable B cell lines expressing Myc-His tagged NS5A. Immunoprecipitation study in the presence or absence of pervanadate (PV implied that NS5A was tyrosine phosphorylated by pervanadate (PV treatment of the cells. Therefore we examined pull-down assay by using glutathione S-transferase (GST-fusion proteins of various Src homology 2 (SH2 domains, which associates with phosphotyrosine within a specific amino acid sequence. The results showed that NS5A specifically bound to SH2 domain of Fyn from PV-treated B cells in addition to Src homology 3 (SH3 domain. Substitution of Arg(176 to Lys in the SH2 domain of Fyn abrogated this interaction. Deletion mutational analysis demonstrated that N-terminal region of NS5A was not required for the interaction with the SH2 domain of Fyn. Tyr(334 was identified as a tyrosine phosphorylation site in NS5A. Far-western analysis revealed that SH2 domain of Fyn directly bound to NS5A. Fyn and NS5A were colocalized in the lipid raft. These results suggest that NS5A directly binds to the SH2 domain of Fyn in a tyrosine phosphorylation-dependent manner. Lastly, we showed that the expression of NS5A in B cells increased phosphorylation of activation loop tyrosine in the kinase domain of Fyn. NS5A containing ligand for both SH2 and SH3 domains enhances an aberrant autophosphorylation and kinase activity of Fyn in B cells.

Data Provenance and Trust

Directory of Open Access Journals (Sweden)

Stratis D Viglas

2013-07-01

Full Text Available The Oxford Dictionary defines provenance as “the place of origin, or earliest known history of something.” The term, when transferred to its digital counterpart, has morphed into a more general meaning. It is not only used to refer to the origin of a digital artefact but also to its changes over time. By changes in this context we may not only refer to its digital snapshots but also to the processes that caused and materialised the change. As an example, consider a database record r created at point in time t0; an update u to that record at time t1 causes it to have a value r’. In terms of provenance, we do not only want to record the snapshots (t0, r and (t1, r’ but also the transformation u that when applied to (t0, r results in (t1, r’, that is u(t0, r = (t1, r’.

TGFbeta Induces Binucleation/Polyploidization in Hepatocytes through a Src-Dependent Cytokinesis Failure.

Science.gov (United States)

De Santis Puzzonia, Marco; Cozzolino, Angela Maria; Grassi, Germana; Bisceglia, Francesca; Strippoli, Raffaele; Guarguaglini, Giulia; Citarella, Franca; Sacchetti, Benedetto; Tripodi, Marco; Marchetti, Alessandra; Amicone, Laura

2016-01-01

In all mammals, the adult liver shows binucleated as well as mononucleated polyploid hepatocytes. The hepatic polyploidization starts after birth with an extensive hepatocyte binucleation and generates hepatocytes of several ploidy classes. While the functional significance of hepatocyte polyploidy is becoming clearer, how it is triggered and maintained needs to be clarified. Aim of this study was to identify a major inducer of hepatocyte binucleation/polyploidization and the cellular and molecular mechanisms involved. We found that, among several cytokines analyzed, known to be involved in early liver development and/or mass control, TGFbeta1 was capable to induce, together with the expected morphological changes, binucleation in hepatocytes in culture. Most importantly, the pharmacological inhibition of TGFbeta signaling in healthy mice during weaning, when the physiological binucleation occurs, induced a significant decrease of hepatocyte binucleation rate, without affecting cell proliferation and hepatic index. The TGFbeta-induced hepatocyte binucleation resulted from a cytokinesis failure, as assessed by video microscopy, and is associated with a delocalization of the cytokinesis regulator RhoA-GTPase from the mid-body of dividing cells. The use of specific chemical inhibitors demonstrated that the observed events are Src-dependent. Finally, the restoration of a fully epithelial phenotype by TGFbeta withdrawal gave rise to a cell progeny capable to maintain the polyploid state. In conclusion, we identified TGFbeta as a major inducer of hepatocyte binucleation both in vitro and in vivo, thus ascribing a novel role to this pleiotropic cytokine. The production of binucleated/tetraploid hepatocytes is due to a cytokinesis failure controlled by the molecular axis TGFbeta/Src/RhoA.

Astrocyte-to-neuron communication through integrin-engaged Thy-1/CBP/Csk/Src complex triggers neurite retraction via the RhoA/ROCK pathway.

Science.gov (United States)

Maldonado, H; Calderon, C; Burgos-Bravo, F; Kobler, O; Zuschratter, W; Ramirez, O; Härtel, S; Schneider, P; Quest, A F G; Herrera-Molina, R; Leyton, L

2017-02-01

Two key proteins for cellular communication between astrocytes and neurons are αvβ3 integrin and the receptor Thy-1. Binding of these molecules in the same (cis) or on adjacent (trans) cellular membranes induces Thy-1 clustering, triggering actin cytoskeleton remodeling. Molecular events that could explain how the Thy-1-αvβ3 integrin interaction signals have only been studied separately in different cell types, and the detailed transcellular communication and signal transduction pathways involved in neuronal cytoskeleton remodeling remain unresolved. Using biochemical and genetic approaches, single-molecule tracking, and high-resolution nanoscopy, we provide evidence that upon binding to αvβ3 integrin, Thy-1 mobility decreased while Thy-1 nanocluster size increased. This occurred concomitantly with inactivation and exclusion of the non-receptor tyrosine kinase Src from the Thy-1/C-terminal Src kinase (Csk)-binding protein (CBP)/Csk complex. The Src inactivation decreased the p190Rho GTPase activating protein phosphorylation, promoting RhoA activation, cofilin, and myosin light chain II phosphorylation and, consequently, neurite shortening. Finally, silencing the adaptor CBP demonstrated that this protein was a key transducer in the Thy-1 signaling cascade. In conclusion, these data support the hypothesis that the Thy-1-CBP-Csk-Src-RhoA-ROCK axis transmitted signals from astrocytic integrin-engaged Thy-1 (trans) to the neuronal actin cytoskeleton. Importantly, the β3 integrin in neurons (cis) was not found to be crucial for neurite shortening. This is the first study to detail the signaling pathway triggered by αvβ3, the endogenous Thy-1 ligand, highlighting the role of membrane-bound integrins as trans acting ligands in astrocyte-neuron communication. Copyright © 2016 Elsevier B.V. All rights reserved.

Autocrine VEGF and IL-8 Promote Migration via Src/Vav2/Rac1/PAK1 Signaling in Human Umbilical Vein Endothelial Cells.

Science.gov (United States)

Ju, Li; Zhou, Zhiwen; Jiang, Bo; Lou, Yue; Guo, Xirong

2017-01-01

Pro-angiogenic factors VEGF and IL-8 play a major role in modulating the migratory potential of endothelial cells. The goal of this study was to investigate the effect of autocrine VEGF and IL-8 in the form of self-conditioned medium (CM) on human umbilical vein endothelial cells (HUVECs). Enzyme-linked immunosorbent assay (ELISA) examined the automatic secretion of VEGF and IL-8 protein by HUVECs. Western blot, small interfering RNA (siRNA), pulldown and Transwell assays were used to explore the role and the mechanism of autocrine VEGF and IL-8 in migration of HUVECs. Neutralizing VEGF and IL-8 in CM significantly abrogated CM-induced migration of HUVECs. Autocrine VEGF and IL-8 increased Src phosphorylation, Rac1 activity and PAK1 phosphorylation in a time dependent manner. Additionally, blocking Rac1 activity with Rac1 siRNA largely abolished autocrine VEGF and IL-8-induced cell migration. Vav2 siRNA suppressed autocrine VEGF and IL-8-induced Rac1 activation and cell migration. Furthermore, blocking Src signaling with PP2, a specific inhibitor for Src, markedly prevented autocrine VEGF and IL-8-induced Vav2 and Rac1 activation as well as consequently cell migration. PAK1 siRNA also significantly abolished autocrine VEGF and IL-8-induced cell migration. We demonstrated for the first time that autocrine VEGF and IL-8 promoted endothelial cell migration via the Src/Vav2/Rac1/PAK1 signaling pathway. This finding reveals the molecular mechanism in the increase of endothelial cell migration induced by autocrine growth factors and cytokines, which is expected to provide a novel therapeutic target in vascular diseases. © 2017 The Author(s)Published by S. Karger AG, Basel.

Crystal structure of the Src family kinase Hck SH3-SH2 linker regulatory region supports an SH3-dominant activation mechanism.

Science.gov (United States)

Alvarado, John J; Betts, Laurie; Moroco, Jamie A; Smithgall, Thomas E; Yeh, Joanne I

2010-11-12

Most mammalian cell types depend on multiple Src family kinases (SFKs) to regulate diverse signaling pathways. Strict control of SFK activity is essential for normal cellular function, and loss of kinase regulation contributes to several forms of cancer and other diseases. Previous x-ray crystal structures of the SFKs c-Src and Hck revealed that intramolecular association of their Src homology (SH) 3 domains and SH2 kinase linker regions has a key role in down-regulation of kinase activity. However, the amino acid sequence of the Hck linker represents a suboptimal ligand for the isolated SH3 domain, suggesting that it may form the polyproline type II helical conformation required for SH3 docking only in the context of the intact structure. To test this hypothesis directly, we determined the crystal structure of a truncated Hck protein consisting of the SH2 and SH3 domains plus the linker. Despite the absence of the kinase domain, the structures and relative orientations of the SH2 and SH3 domains in this shorter protein were very similar to those observed in near full-length, down-regulated Hck. However, the SH2 kinase linker adopted a modified topology and failed to engage the SH3 domain. This new structure supports the idea that these noncatalytic regions work together as a "conformational switch" that modulates kinase activity in a manner unique to the SH3 domain and linker topologies present in the intact Hck protein. Our results also provide fresh structural insight into the facile induction of Hck activity by HIV-1 Nef and other Hck SH3 domain binding proteins and implicate the existence of innate conformational states unique to individual Src family members that "fine-tune" their sensitivities to activation by SH3-based ligands.

Big Data Provenance: Challenges, State of the Art and Opportunities

OpenAIRE

Wang, Jianwu; Crawl, Daniel; Purawat, Shweta; Nguyen, Mai; Altintas, Ilkay

2015-01-01

Ability to track provenance is a key feature of scientific workflows to support data lineage and reproducibility. The challenges that are introduced by the volume, variety and velocity of Big Data, also pose related challenges for provenance and quality of Big Data, defined as veracity. The increasing size and variety of distributed Big Data provenance information bring new technical challenges and opportunities throughout the provenance lifecycle including recording, querying, sharing and ut...

Catalytic Filtration: A proven technology for Dioxin emission control from waste incinerators

International Nuclear Information System (INIS)

Wong, K.T.; Xu, Zhengtian

2010-01-01

Polychlorinated dibenzo-p-dioxins and di benzofurans (PCD/ Fs), in a family of dioxin derivatives with high toxicity, often associated with environmental pollution are the most toxic man made substances, emitted in gas and solid phases during incineration of waste. The threat of dioxin is drawing increasing attention around the world. Governments around the world are phasing in more stringent dioxin emission regulations, and reports about dioxin levels in food products have generated widespread concerns among the public. Issues related to dioxin emissions and disposals are moving up the environmental agenda demanding the most effective and environmentally sound technologies. With heightened public awareness, more stringent regulations, and potential penalties for non-compliance, its more important than ever to avoid the risks associated with inadequate dioxin control. The permissible dioxin emission in most industrial nations is less than 0.1 ng (TEQ)/ Nm 3 and permissible dust emission is from less than 10 to less than 50 mg/ Nm 3 . The common system to remove dioxin is installing an injection process for powdered activated carbon (PAC). This was seen as a proven and widely used technology to control dioxin. This sorbent based system moves dioxin and furan molecules from the gas stream to the solid residue. There are new concerns about existing or future landfill restrictions on the amount and toxicity of sorbent levels in fly ash. Other alternatives are non-flammable additives and catalytic technologies. The non-flammable additives are not proven to control dioxin at temperatures above 200 degree Celsius. Catalytic filter technology can be high initial investment but gaining popularity for operational benefits and reduction of solid residues for landfill. Several criteria are being considered to compare the initial cost of the catalytic filter system and the cost reduction of exhaust gas treatment that can pay for the return of the investment. Field experiences

Innocent Until Proven Guilty

Science.gov (United States)

Case, Catherine; Whitaker, Douglas

2016-01-01

In the criminal justice system, defendants accused of a crime are presumed innocent until proven guilty. Statistical inference in any context is built on an analogous principle: The null hypothesis--often a hypothesis of "no difference" or "no effect"--is presumed true unless there is sufficient evidence against it. In this…

Nonredundant roles of Src-family kinases and Syk in the initiation of B-cell antigen receptor signaling

Czech Academy of Sciences Publication Activity Database

Štěpánek, Ondřej; Dráber, Peter; Drobek, Aleš; Hořejší, Václav; Brdička, Tomáš

2013-01-01

Roč. 190, č. 4 (2013), s. 1807-1818 ISSN 0022-1767 R&D Projects: GA ČR(CZ) GBP302/12/G101; GA ČR GAP302/12/1712 Institutional support: RVO:68378050 Keywords : BCR signaling * Src family kinases * Syk Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 5.362, year: 2013

Regulation of Src family kinases involved in T cell receptor signaling by protein-tyrosine phosphatase CD148

Czech Academy of Sciences Publication Activity Database

Štěpánek, Ondřej; Kalina, T.; Dráber, Peter; Skopcová, Tereza; Svojgr, K.; Angelisová, Pavla; Hořejší, Václav; Weiss, A.; Brdička, Tomáš

2011-01-01

Roč. 286, č. 25 (2011), s. 22101-22112 ISSN 0021-9258 R&D Projects: GA MŠk 2B06064; GA MŠk 1M0506 Institutional research plan: CEZ:AV0Z50520514 Keywords : CD148 * tyrosine phosphatase * Src family kinases Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 4.773, year: 2011

ZINC-INDUCED EGF RECEPTOR SIGNALING REQUIRES SRC-MEDIATED PHOSPHORYLATION OF THE EGF RECEPTOR ON TYROSINE 845 (Y845)

Science.gov (United States)

ZINC-INDUCED EGF RECEPTOR SIGNALING REQUIRES Src-MEDIATED PHOSPHORYLATION OF THE EGF RECEPTOR ON TYROSINE 845 (Y845)Weidong Wu1, Lee M. Graves2, Gordon N. Gill3 and James M. Samet4 1Center for Environmental Medicine and Lung Biology; 2Department of Pharmacology, University o...

Paeonol Suppresses Chondrosarcoma Metastasis through Up-Regulation of miR-141 by Modulating PKCδ and c-Src Signaling Pathway

Science.gov (United States)

Horng, Chi-Ting; Shieh, Po-Chuen; Tan, Tzu-Wei; Yang, Wei-Hung; Tang, Chih-Hsin

2014-01-01

Chondrosarcoma, a primary malignant bone cancer, has potential for local invasion and distant metastasis, especially to the lungs. Patients diagnosed with it show poor prognosis. Paeonol (2'-hydroxy-4'-methoxyacetophenone), the main active compound of traditional Chinese remedy Paeonia lactiflora Pallas, exhibits anti-inflammatory and anti-tumor activity; whether paeonol regulates metastatic chondrosarcoma is largely unknown. Here, we find paeonol do not increase apoptosis. By contrast, at non-cytotoxic concentrations, paeonol suppresses migration and invasion of chondrosarcoma cells. We also demonstrate paeonol enhancing miR-141 expression and miR-141 inhibitor reversing paeonol-inhibited cell motility; paeonol also reduces protein kinase C (PKC)δ and c-Src kinase activity. Since paeonol inhibits migration and invasion of human chondrosarcoma via up-regulation of miR-141 via PKCδ and c-Src pathways, it thus might be a novel anti-metastasis agent for treatment of metastatic chondrosarcoma. PMID:24992595

SRC-willow (Salix viminalis) as a resource for flower-visiting insects

Energy Technology Data Exchange (ETDEWEB)

Reddersen, J. [National Environmental Research Institute, Ronde (Denmark). Dept. of Landscape Ecology

2001-07-01

The potential habitat value of commercial short rotation coppice (SRC)-willow plantations for flower-visiting insects was investigated. During 1998-2000, at a single typical intensive Danish farmland site, 11 Salix viminalis plantations were sampled by late April to quantify willow catkin abundance and flower sex. Mean plantation size was 1.1 ha and included one or more of clones: orm, rapp, ulv, jorr, christina and jorrun. Plot-year means of catkin abundance and of proportion of willows flowering were related to the coppicing cycle, i.e. the number of growth years since last harvest of plot ('year' 0-4). In 1998, the ground layer vegetation was sampled. Monitoring flower-visiting insects by means of line-transect counts failed due to the local scarcity of bees. At the plantation scale, flowering was discontinuous across the harvest cycle as it was totally absent in the year immediately following harvest. In successive years (1-4), individual willows flowered frequently and, occasionally, at high abundances, and catkin abundance increased with time. Within 3-4 year of harvest cycle, all plots flowered in most years with most plots exhibiting at least some flowering in any 1 year. Thus, willow catkin abundance was generally high in the total area due to: high frequency of flowering in plots, occasional high flowering abundance, plots not being harvested simultaneously and large total number of willows within plots and landscape. Similarly, flower sex ratio, and thus flower value, varied greatly between plots while variation was damped across plots. Alternative simultaneous flower resources in ground layer vegetation were few except for Dandelion. SRC willow may constitute an important resource for bees, even under the stress of the harvest cycle, and recommendations are given for improving this biodiversity aspect. (author)

Tracing where and who provenance in Linked Data: A calculus

OpenAIRE

Dezani-Ciancaglini, Mariangiola; Horne, Ross; Sassone, Vladimiro

2012-01-01

Linked Data provides some sensible guidelines for publishing and consuming data on the Web. Data published on the Web has no inherent truth, yet its quality can often be assessed based on its provenance. This work introduces a new approach to provenance for Linked Data. The simplest notion of provenance-viz., a named graph indicating where the data is now-is extended with a richer provenance format. The format reflects the behaviour of processes interacting with Linked Data, tracing where the...

Restful Implementation of Catalogue Service for Geospatial Data Provenance

Science.gov (United States)

Jiang, L. C.; Yue, P.; Lu, X. C.

2013-10-01

Provenance, also known as lineage, is important in understanding the derivation history of data products. Geospatial data provenance helps data consumers to evaluate the quality and reliability of geospatial data. In a service-oriented environment, where data are often consumed or produced by distributed services, provenance could be managed by following the same service-oriented paradigm. The Open Geospatial Consortium (OGC) Catalogue Service for the Web (CSW) is used for the registration and query of geospatial data provenance by extending ebXML Registry Information Model (ebRIM). Recent advance of the REpresentational State Transfer (REST) paradigm has shown great promise for the easy integration of distributed resources. RESTful Web Service aims to provide a standard way for Web clients to communicate with servers based on REST principles. The existing approach for provenance catalogue service could be improved by adopting the RESTful design. This paper presents the design and implementation of a catalogue service for geospatial data provenance following RESTful architecture style. A middleware named REST Converter is added on the top of the legacy catalogue service to support a RESTful style interface. The REST Converter is composed of a resource request dispatcher and six resource handlers. A prototype service is developed to demonstrate the applicability of the approach.

Valorization of agroindustrial solid residues and residues from biofuel production chains by thermochemical conversion: a review, citing Brazil as a case study

Directory of Open Access Journals (Sweden)

E. Virmond

2013-06-01

Full Text Available Besides high industrial development, Brazil is also an agribusiness country. Each year about 330 million metrics tons (Mg of biomass residues are generated, requiring tremendous effort to develop biomass systems in which production, conversion and utilization of bio-based products are carried out efficiently and under environmentally sustainable conditions. For the production of biofuels, organic chemicals and materials, it is envisaged to follow a biorefinery model which includes modern and proven green chemical technologies such as bioprocessing, pyrolysis, gasification, Fischer-Tropsch synthesis and other catalytic processes in order to make more complex molecules and materials on which a future sustainable society will be based. This paper presents promising options for valorization of Brazilian agroindustrial biomass sources and residues originating from the biofuel production chains as renewable energy sources and addresses the main aspects of the thermochemical technologies which have been applied.

2D QSAR studies of the inhibitory activity of a series of substituted purine derivatives against c-Src tyrosine kinase

Directory of Open Access Journals (Sweden)

Mukesh C. Sharma

2016-07-01

Full Text Available A series of 34 substituted purine analogues derivatives were subjected to quantitative structure-activity relationship analyses as inhibitors of c-Src tyrosine kinase. Partial least squares regression was applied to derive QSAR models, which were further validated for statistical significance by internal and external validation. The best QSAR model developed had a good predictive correlation coefficient (r2 of 0.8319, a significant cross-validated correlation coefficient (q2 of 0.7550, and an r2 for the external test set (pred_r2 of 0.7983. It was developed from the PLS method with descriptors including the SsCH3E-index, H-Donor Count, T_2_Cl_3, and negative correlation with SsOHcount. The current study provides better insight into the future design of more potent c-Src tyrosine kinase inhibitors prior to synthesis.

Growth and provenance variation of Pinus caribaea var ...

African Journals Online (AJOL)

CAMCORE has visited 33 populations of Pinus caribaea var. hondurensis in Belize, Guatemala, Honduras, El Salvador, Nicaragua, and Quintana Roo, Mexico. Seed collections have been made in 29 provenances from 1, 325 mother trees. A total of 21 provenances and sources of Pinus caribaea var. hondurensis were ...

Heterozygous Null Bone Morphogenetic Protein Receptor Type 2 Mutations Promote SRC Kinase-dependent Caveolar Trafficking Defects and Endothelial Dysfunction in Pulmonary Arterial Hypertension*

Science.gov (United States)

Prewitt, Allison R.; Ghose, Sampa; Frump, Andrea L.; Datta, Arumima; Austin, Eric D.; Kenworthy, Anne K.; de Caestecker, Mark P.

2015-01-01

Hereditary pulmonary arterial hypertension (HPAH) is a rare, fatal disease of the pulmonary vasculature. The majority of HPAH patients inherit mutations in the bone morphogenetic protein type 2 receptor gene (BMPR2), but how these promote pulmonary vascular disease is unclear. HPAH patients have features of pulmonary endothelial cell (PEC) dysfunction including increased vascular permeability and perivascular inflammation associated with decreased PEC barrier function. Recently, frameshift mutations in the caveolar structural protein gene Caveolin-1 (CAV-1) were identified in two patients with non-BMPR2-associated HPAH. Because caveolae regulate endothelial function and vascular permeability, we hypothesized that defects in caveolar function might be a common mechanism by which BMPR2 mutations promote pulmonary vascular disease. To explore this, we isolated PECs from mice carrying heterozygous null Bmpr2 mutations (Bmpr2+/−) similar to those found in the majority of HPAH patients. We show that Bmpr2+/− PECs have increased numbers and intracellular localization of caveolae and caveolar structural proteins CAV-1 and Cavin-1 and that these defects are reversed after blocking endocytosis with dynasore. SRC kinase is also constitutively activated in Bmpr2+/− PECs, and localization of CAV-1 to the plasma membrane is restored after treating Bmpr2+/− PECs with the SRC kinase inhibitor 3-(4-chlorophenyl)-1-(1,1-dimethylethyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (PP2). Late outgrowth endothelial progenitor cells isolated from HPAH patients show similar increased activation of SRC kinase. Moreover, Bmpr2+/− PECs have impaired endothelial barrier function, and barrier function is restored after treatment with PP2. These data suggest that heterozygous null BMPR2 mutations promote SRC-dependent caveolar trafficking defects in PECs and that this may contribute to pulmonary endothelial barrier dysfunction in HPAH patients. PMID:25411245

Hydrotreating catalyst deactivation by coke from SRC-II oil

Energy Technology Data Exchange (ETDEWEB)

Yamamoto, Y.; Kumata, F.; Massoth, F.E.

1988-10-01

Samples of a CoMo/Al/sub 2/O/sub 3/ catalyst were partially deactivated with SRC-II feed in an autoclave reactor to give coked samples of 5 to 18% C. The coked catalysts were analyzed for surface area, pore volume, coronene adsorption and diffusivity, and their catalytic activity determined for hydrodesulfurization (HDS), hydrodeoxygenation (HDO) and C-N hydrogenolysis (CNH) using model compounds. All of the above measurements decreased with increase in coke content. Property data indicate that some pores are blocked by coke and diffusivity results show narrowing of pore mouths with increasing coke content. Catalyst deactivation versus coke level was identical for HDS and HDO, but less for CNH. A simple model of coke deactivation was developed to relate activity to coke content. Coke is envisioned as forming wedge-like deposits in the catalyst pores. 11 refs., 5 figs., 3 tabs.

Src Is a Prime Target Inhibited by Celtis choseniana Methanol Extract in Its Anti-Inflammatory Action

Directory of Open Access Journals (Sweden)

Han Gyung Kim

2018-01-01

Full Text Available Celtis choseniana is the traditional plant used at Korea as a herbal medicine to ameliorate inflammatory responses. Although Celtis choseniana has been traditionally used as a herbal medicine at Korea, no systemic research has been conducted on its anti-inflammatory activity. Therefore, the present study explored an anti-inflammatory effect and its underlying molecular mechanism using Celtis choseniana methanol extract (Cc-ME in macrophage-mediated inflammatory responses. In vitro anti-inflammatory activity of Cc-ME was evaluated using RAW264.7 cells and peritoneal macrophages stimulated by lipopolysaccharide (LPS, pam3CSK4 (Pam3, or poly(I:C. In vivo anti-inflammatory activity of Cc-ME was investigated using acute inflammatory disease mouse models, such as LPS-induced peritonitis and HCl/EtOH-induced gastritis. The molecular mechanism of Cc-ME-mediated anti-inflammatory activity was examined by Western blot analysis and immunoprecipitation using whole cell and nuclear fraction prepared from the LPS-stimulated RAW264.7 cells and HEK293 cells. Cc-ME inhibited NO production and mRNA expression of inducible nitric oxide synthase (iNOS, cyclooxygenase (COX-2, and tumor necrosis factor-alpha (TNF-α in the RAW264.7 cells and peritoneal macrophages induced by LPS, pam3, or poly(I:C without cytotoxicity. High-performance liquid chromatography (HPLC analysis showed that Cc-ME contained anti-inflammatory flavonoids quercetin, luteolin, and kaempferol. Among those, the content of luteolin, which showed an inhibitory effect on NO production, was highest. Cc-ME suppressed the NF-κB signaling pathway by targeting Src and interrupting molecular interactions between Src and p85, its downstream kinase. Moreover, Cc-ME ameliorated the morphological finding of peritonitis and gastritis in the mouse disease models. Therefore, these results suggest that Cc-ME exerted in vitro and in vivo anti-inflammatory activity in LPS-stimulated macrophages and mouse models of

TGFbeta Induces Binucleation/Polyploidization in Hepatocytes through a Src-Dependent Cytokinesis Failure.

Directory of Open Access Journals (Sweden)

Marco De Santis Puzzonia

Full Text Available In all mammals, the adult liver shows binucleated as well as mononucleated polyploid hepatocytes. The hepatic polyploidization starts after birth with an extensive hepatocyte binucleation and generates hepatocytes of several ploidy classes. While the functional significance of hepatocyte polyploidy is becoming clearer, how it is triggered and maintained needs to be clarified. Aim of this study was to identify a major inducer of hepatocyte binucleation/polyploidization and the cellular and molecular mechanisms involved. We found that, among several cytokines analyzed, known to be involved in early liver development and/or mass control, TGFbeta1 was capable to induce, together with the expected morphological changes, binucleation in hepatocytes in culture. Most importantly, the pharmacological inhibition of TGFbeta signaling in healthy mice during weaning, when the physiological binucleation occurs, induced a significant decrease of hepatocyte binucleation rate, without affecting cell proliferation and hepatic index. The TGFbeta-induced hepatocyte binucleation resulted from a cytokinesis failure, as assessed by video microscopy, and is associated with a delocalization of the cytokinesis regulator RhoA-GTPase from the mid-body of dividing cells. The use of specific chemical inhibitors demonstrated that the observed events are Src-dependent. Finally, the restoration of a fully epithelial phenotype by TGFbeta withdrawal gave rise to a cell progeny capable to maintain the polyploid state. In conclusion, we identified TGFbeta as a major inducer of hepatocyte binucleation both in vitro and in vivo, thus ascribing a novel role to this pleiotropic cytokine. The production of binucleated/tetraploid hepatocytes is due to a cytokinesis failure controlled by the molecular axis TGFbeta/Src/RhoA.

Regulation of mTORC1 signaling by Src kinase activity is Akt1-independent in RSV-transformed cells

Czech Academy of Sciences Publication Activity Database

Vojtěchová, Martina; Turečková, Jolana; Kučerová, Dana; Šloncová, Eva; Vachtenheim, J.; Tuháčková, Zdena

2008-01-01

Roč. 10, č. 2 (2008), s. 99-107 ISSN 1522-8002 R&D Projects: GA ČR GA301/04/0550 Institutional research plan: CEZ:AV0Z50520514 Keywords : Akt/PKB * mTOR C1 signaling pathway * Src Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 5.191, year: 2008

Clonorchis sinensis excretory-secretory products promote the migration and invasion of cholangiocarcinoma cells by activating the integrin β4-FAK/Src signaling pathway.

Science.gov (United States)

Pak, Jhang Ho; Bashir, Qudsia; Kim, In Ki; Hong, Sung-Jong; Maeng, Sejung; Bahk, Young Yil; Kim, Tong-Soo

2017-06-01

Cholangiocarcinoma (CCA) is a slow-growing but highly metastatic cancer. Its metastatic potential largely explains its high mortality rate. A recognized risk factor for CCA development is infection with the liver flukes Opisthorchis viverrini and Clonorchis sinensis. We previously reported that the excretory-secretory products (ESPs) of C. sinensis promoted the three-dimensional aggregation and invasion of CCA cells. In the present study, a quantitative real-time PCR array of extracellular matrix (ECM) and adhesion molecules was used to examine the regulatory mechanism of ESP-mediated CCA cell migration and invasion. In particular, the expression levels of integrin α isoforms and β4 were upregulated in response to ESPs. Increased expression of integrin β4 was probably correlated with activation of focal adhesion kinase (FAK) and the steroid receptor coactivator (Src) family kinase and the subsequent activation of two downstream focal adhesion molecules, paxillin and vinculin. Moreover, inhibition of FAK/Src activation reduced paxillin and vinculin phosphorylation and attenuated ESP-induced CCA cell migration and invasion. These findings suggest that the integrin β4-FAK/Src signaling axis may play a crucial role in clonorchiasis-associated CCA metastasis during tumor progression. Copyright © 2017 Elsevier B.V. All rights reserved.

Querying Provenance Information: Basic Notions and an Example from Paleoclimate Reconstruction

Science.gov (United States)

Stodden, V.; Ludaescher, B.; Bocinsky, K.; Kintigh, K.; Kohler, T.; McPhillips, T.; Rush, J.

2016-12-01

Computational models are used to reconstruct and explain past environments and to predict likely future environments. For example, Bocinsky and Kohler have performed a 2,000-year reconstruction of the rain-fed maize agricultural niche in the US Southwest. The resulting academic publications not only contain traditional method descriptions, figures, etc. but also links to code and data for basic transparency and reproducibility. Examples include ResearchCompendia.org and the new project "Merging Science and Cyberinfrastructure Pathways: The Whole Tale." Provenance information provides a further critical element to understand a published study and to possibly extend or challenge the findings of the original authors. We present different notions and uses of provenance information using a computational archaeology example, e.g., the common use of "provenance for others" (for transparency and reproducibility), but also the more elusive but equally important use of "provenance for self". To this end, we distinguish prospective provenance (a.k.a. workflow) from retrospective provenance (a.k.a. data lineage) and show how combinations of both forms of provenance can be used to answer different kinds of important questions about a workflow and its execution. Since many workflows are developed using scripting or special purpose languages such as Python and R, we employ an approach and toolkit called YesWorkflow that brings provenance modeling, capture, and querying into the realm of scripting. YesWorkflow employs the basic W3C PROV standard, as well as the ProvONE extension for sharing and exchanging retrospective and prospective provenance information, respectively. Finally, we argue that the utility of provenance information should be maximized by developing different kinds provenance questions and queries during the early phases of computational workflow design and implementation.

Provenance variation in Eucalyptus camaldulensis Dehnh. in California

Science.gov (United States)

B.M. Emery; F. Thomas Ledig

1987-01-01

In California, the Lake Albacutya provenance of river red gum was clearly superior in volume growth to 22 other provenances collected throughout the range of the species in Australia. It had at least 2.5 times the volume of the plantation mean at 5.5 years, consistent with its performance in other countries with Mediterranean climates like California's. Other...

Diverging Drought Resistance of Scots Pine Provenances Revealed by Infrared Thermography

Directory of Open Access Journals (Sweden)

Hannes Seidel

2016-08-01

Full Text Available With recent climate changes, Scots pine (Pinus sylvestris L. forests have been affected by die-off events. Assisted migration of adapted provenances mitigates drought impacts and promotes forest regeneration. Although suitable provenances are difficult to identify by traditional ecophysiological techniques, which are time consuming and invasive, plant water status can be easily assessed by infrared thermography. Thus, we examined the stress responses of 2-year-old potted Scots pine seedlings from six provenances (Bulgaria, France, Germany, Italy, Poland, and Spain based on two thermal indices (crop water stress index and stomatal conductance index. Both indices were derived from infrared images during a six-week drought/control treatment in a greenhouse in the summer of 2013. The pines were monitored during the stress and subsequent recovery period. After controlling for fluctuating environmental conditions, soil moisture or treatment-specific water supply was the most important driver of drought stress. The stress magnitude and response to soil water deficit depended on provenance. Under moderate drought conditions, pines from western and eastern Mediterranean provenances (Bulgaria, France, and Spain expressed lower stress levels than those from both continental provenances (Germany and Poland. In pines from the Spanish and Bulgarian provenances, the stress level differences were significantly lower than in continental pines. Moreover, pines from continental provenances were less resilient (showed less recovery after the stress period than Mediterranean pines. Under extreme drought, all provenances were equally stressed with almost no significant differences in their thermal indices. Provenance-specific differences in drought resistance, which are associated with factors such as summer precipitation at the origin of Scots pine seedlings, may offer promising tracks of adaptation to future drought risks.

P120-GAP associated with syndecan-2 to function as an active switch signal for Src upon transformation with oncogenic ras

International Nuclear Information System (INIS)

Huang, J.-W.; Chen, C.-L.; Chuang, N.-N.

2005-01-01

BALB/3T3 cells transfected with plasmids pcDNA3.1-[S-ras(Q 61 K)] of shrimp Penaeus japonicus were applied to reveal a complex of p120-GAP/syndecan-2 being highly expressed upon transformation. Of interest, most of the p120-GAP/syndecan-2 complex was localized at caveolae, a membrane microdomain enriched with caveolin-1. To confirm the molecular interaction between syndecan-2 and p120-GAP, we further purified p120-GAP protein from mouse brains by using an affinity column of HiTrap-RACK1 and expressed mouse RACK1-encoded fusion protein and mouse syndecan-2-encoded fusion protein in bacteria. We report molecular affinities exist between p120-GAP and RACK1, syndecan-2 and RACK1 as well as p120-GAP and syndecan-2. The selective affinity between p120-GAP and syndecan-2 was found to be sufficient to detach RACK1. The p120-GAP/syndecan-2 complex was demonstrated to keep Src tyrosine kinase in an activated form. On the other hand, the syndecan-2/RACK1 complex was found to have Src in an inactivated form. These data indicate that the p120-GAP/syndecan-2 complex at caveolae could provide a docking site for Src to transmit tyrosine signaling, implying that syndecan-2/p120-GAP functions as a tumor promoter upon transformation with oncogenic ras of shrimp P. japonicus

Inhibition of tumor metastasis by a growth factor receptor bound protein 2 Src homology 2 domain-binding antagonist.

Science.gov (United States)

Giubellino, Alessio; Gao, Yang; Lee, Sunmin; Lee, Min-Jung; Vasselli, James R; Medepalli, Sampath; Trepel, Jane B; Burke, Terrence R; Bottaro, Donald P

2007-07-01

Metastasis, the primary cause of death in most forms of cancer, is a multistep process whereby cells from the primary tumor spread systemically and colonize distant new sites. Blocking critical steps in this process could potentially inhibit tumor metastasis and dramatically improve cancer survival rates; however, our understanding of metastasis at the molecular level is still rudimentary. Growth factor receptor binding protein 2 (Grb2) is a widely expressed adapter protein with roles in epithelial cell growth and morphogenesis, as well as angiogenesis, making it a logical target for anticancer drug development. We have previously shown that a potent antagonist of Grb2 Src homology-2 domain-binding, C90, blocks growth factor-driven cell motility in vitro and angiogenesis in vivo. We now report that C90 inhibits metastasis in vivo in two aggressive tumor models, without affecting primary tumor growth rate. These results support the potential efficacy of this compound in reducing the metastatic spread of primary solid tumors and establish a critical role for Grb2 Src homology-2 domain-mediated interactions in this process.

Growth and Survival Variation among Scots Pine (Pinus sylvestris L. Provenances

Directory of Open Access Journals (Sweden)

Süleyman Gülcü

2017-01-01

Full Text Available Tree height, basal diameter, and survival were examined in thirteen-year-old provenance test established by 30 seed sources of Scots pine (Pinus sylvestris L. at two exotic sites of the species in Southern part of Turkey. Variations within provenance and among provenances and relations among the traits were estimated to compare Scots pine provenance and two other native species. Averages of tree height and basal diameter were 350 cm and 52.7 mm in Aydogmus site and 385 cm and 51.2 mm in Kemer site, respectively. There were large differences within and among provenances for the characters. Sites were similar (p>0.05 for the characters, while there were significant differences (p≤0.05 among provenances within site according to results of variance analysis (ANOVA. Scots pine provenances were higher and had more thickness than that of black pine (Pinus nigra Arnold and Taurus cedar (Cedrus libani A. Rich. which were natural species of the region. There were positive and significant (p<0.05 correlations between height and basal diameter in the species. Average survivals were 56% and 35% of the provenances in the sites. They were 71% and 11% in black pine and 53% in Taurus cedar for the sites respectively.

Molecular characterization of c-Abl/c-Src kinase inhibitors targeted against murine tumour progenitor cells that express stem cell markers.

Directory of Open Access Journals (Sweden)

Thomas Kruewel

Full Text Available BACKGROUND: The non-receptor tyrosine kinases c-Abl and c-Src are overexpressed in various solid human tumours. Inhibition of their hyperactivity represents a molecular rationale in the combat of cancerous diseases. Here we examined the effects of a new family of pyrazolo [3,4-d] pyrimidines on a panel of 11 different murine lung tumour progenitor cell lines, that express stem cell markers, as well as on the human lung adenocarcinoma cell line A549, the human hepatoma cell line HepG2 and the human colon cancer cell line CaCo2 to obtain insight into the mode of action of these experimental drugs. METHODOLOGY/PRINCIPAL FINDINGS: Treatment with the dual kinase inhibitors blocked c-Abl and c-Src kinase activity efficiently in the nanomolar range, induced apoptosis, reduced cell viability and caused cell cycle arrest predominantly at G0/G1 phase while western blot analysis confirmed repressed protein expression of c-Abl and c-Src as well as the interacting partners p38 mitogen activated protein kinase, heterogenous ribonucleoprotein K, cyclin dependent kinase 1 and further proteins that are crucial for tumour progression. Importantly, a significant repression of the epidermal growth factor receptor was observed while whole genome gene expression analysis evidenced regulation of many cell cycle regulated genes as well integrin and focal adhesion kinase (FAK signalling to impact cytoskeleton dynamics, migration, invasion and metastasis. CONCLUSIONS/SIGNIFICANCE: Our experiments and recently published in vivo engraftment studies with various tumour cell lines revealed the dual kinase inhibitors to be efficient in their antitumour activity.

PDGF-induced migration of synthetic vascular smooth muscle cells through c-Src-activated L-type Ca2+ channels with full-length CaV1.2 C-terminus.

Science.gov (United States)

Guo, Xiaoguang; Kashihara, Toshihide; Nakada, Tsutomu; Aoyama, Toshifumi; Yamada, Mitsuhiko

2018-06-01

In atherosclerosis, vascular smooth muscle cells (VSMC) migrate from the media toward the intima of the arteries in response to cytokines, such as platelet-derived growth factor (PDGF). However, molecular mechanism underlying the PDGF-induced migration of VSMCs remains unclear. The migration of rat aorta-derived synthetic VSMCs, A7r5, in response to PDGF was potently inhibited by a Ca V 1.2 channel inhibitor, nifedipine, and a Src family tyrosine kinase (SFK)/Abl inhibitor, bosutinib, in a less-than-additive manner. PDGF significantly increased Ca V 1.2 channel currents without altering Ca V 1.2 protein expression levels in A7r5 cells. This reaction was inhibited by C-terminal Src kinase, a selective inhibitor of SFKs. In contractile VSMCs, the C-terminus of Ca V 1.2 is proteolytically cleaved into proximal and distal C-termini (PCT and DCT, respectively). Clipped DCT is noncovalently reassociated with PCT to autoinhibit the channel activity. Conversely, in synthetic A7r5 cells, full-length Ca V 1.2 (Ca V 1.2FL) is expressed much more abundantly than truncated Ca V 1.2. In a heterologous expression system, c-Src activated Ca V 1.2 channels composed of Ca V 1.2FL but not truncated Ca V 1.2 (Ca V 1.2Δ1763) or Ca V 1.2Δ1763 plus clipped DCT. Further, c-Src enhanced the coupling efficiency between the voltage-sensing domain and activation gate of Ca V 1.2FL channels by phosphorylating Tyr1709 and Tyr1758 in PCT. Compared with Ca V 1.2Δ1763, c-Src could more efficiently bind to and phosphorylate Ca V 1.2FL irrespective of the presence or absence of clipped DCT. Therefore, in atherosclerotic lesions, phenotypic switching of VSMCs may facilitate pro-migratory effects of PDGF on VSMCs by suppressing posttranslational Ca V 1.2 modifications.

Data Provenance for Agent-Based Models in a Distributed Memory

Directory of Open Access Journals (Sweden)

Delmar B. Davis

2018-04-01

Full Text Available Agent-Based Models (ABMs assist with studying emergent collective behavior of individual entities in social, biological, economic, network, and physical systems. Data provenance can support ABM by explaining individual agent behavior. However, there is no provenance support for ABMs in a distributed setting. The Multi-Agent Spatial Simulation (MASS library provides a framework for simulating ABMs at fine granularity, where agents and spatial data are shared application resources in a distributed memory. We introduce a novel approach to capture ABM provenance in a distributed memory, called ProvMASS. We evaluate our technique with traditional data provenance queries and performance measures. Our results indicate that a configurable approach can capture provenance that explains coordination of distributed shared resources, simulation logic, and agent behavior while limiting performance overhead. We also show the ability to support practical analyses (e.g., agent tracking and storage requirements for different capture configurations.

DataONE: A Data Federation with Provenance Support

Energy Technology Data Exchange (ETDEWEB)

Cao, Yang [University of Illinois at Urbana-Champaign; Jones, Christopher [National Center for Ecological Analysis and Synthesis, Santa Barbara; Cuevas-Vicenttin, Victor [ Universidad Popular Autónoma del Estado de Puebla, Puebla, Mexico; Jones, Matthew B. [National Center for Ecological Analysis and Synthesis, Santa Barbara; Ludascher, Bertram [University of Illinois at Urbana-Champaign; McPhillips, Timothy [University of Illinois at Urbana-Champaign; Missier, Paolo [Newcastle University, UK; Schwalm, Christopher [Woods Hole Research Center (WHRC), Massachusetts; Slaughter, Peter [National Center for Ecological Analysis and Synthesis, Santa Barbara; Vieglais, Dave [University of Kansas; Walker, Lauren [National Center for Ecological Analysis and Synthesis, Santa Barbara

2016-01-01

DataONE is a federated data network focusing on earth and environmental science data. We present the provenance and search features of DataONE by means of an example involving three earth scientists who interact through a DataONE Member Node. DataONE provenance systems enable reproducible research and facilitate proper attribution of scientific results transitively across generations of derived data products.

Polymeric immunoglobulin receptor-mediated invasion of Streptococcus pneumoniae into host cells requires a coordinate signaling of SRC family of protein-tyrosine kinases, ERK, and c-Jun N-terminal kinase.

Science.gov (United States)

Agarwal, Vaibhav; Asmat, Tauseef M; Dierdorf, Nina I; Hauck, Christof R; Hammerschmidt, Sven

2010-11-12

Streptococcus pneumoniae are commensals of the human nasopharynx with the capacity to invade mucosal respiratory cells. PspC, a pneumococcal surface protein, interacts with the human polymeric immunoglobulin receptor (pIgR) to promote bacterial adherence to and invasion into epithelial cells. Internalization of pneumococci requires the coordinated action of actin cytoskeleton rearrangements and the retrograde machinery of pIgR. Here, we demonstrate the involvement of Src protein-tyrosine kinases (PTKs), focal adhesion kinase (FAK), extracellular signal-regulated kinase (ERK), and c-Jun N-terminal kinase (JNK) but not p38 mitogen-activated protein kinases (MAPK) in pneumococcal invasion via pIgR. Pharmacological inhibitors of PTKs and MAPKs and genetic interference with Src PTK and FAK functions caused a significant reduction of pIgR-mediated pneumococcal invasion but did not influence bacterial adhesion to host cells. Furthermore, pneumococcal ingestion by host cells induces activation of ERK1/2 and JNK. In agreement with activated JNK, its target molecule and DNA-binding protein c-Jun was phosphorylated. We also show that functionally active Src PTK is essential for activation of ERK1/2 upon pneumococcal infections. In conclusion, these data illustrate the importance of a coordinated signaling between Src PTKs, ERK1/2, and JNK during PspC-pIgR-mediated uptake of pneumococci by host epithelial cells.

Mobility of TOAC spin-labelled peptides binding to the Src SH3 domain studied by paramagnetic NMR

Energy Technology Data Exchange (ETDEWEB)

Lindfors, Hanna E. [Leiden University, Leiden Institute of Chemistry, Gorlaeus Laboratories (Netherlands); Koning, Peter E. de; Wouter Drijfhout, Jan [Leiden University Medical Centre, Department of Immunohematology and Blood Transfusion (Netherlands); Venezia, Brigida; Ubbink, Marcellus [Leiden University, Leiden Institute of Chemistry, Gorlaeus Laboratories (Netherlands)], E-mail: m.ubbink@chem.leidenuniv.nl

2008-07-15

Paramagnetic relaxation enhancement provides a tool for studying the dynamics as well as the structure of macromolecular complexes. The application of side-chain coupled spin-labels is limited by the mobility of the free radical. The cyclic, rigid amino acid spin-label TOAC (2,2,6,6-Tetramethylpiperidine-1-oxyl-4-amino-4-carboxylic acid), which can be incorporated straightforwardly by peptide synthesis, provides an attractive alternative. In this study, TOAC was incorporated into a peptide derived from focal adhesion kinase (FAK), and the interaction of the peptide with the Src homology 3 (SH3) domain of Src kinase was studied, using paramagnetic NMR. Placing TOAC within the binding motif of the peptide has a considerable effect on the peptide-protein binding, lowering the affinity substantially. When the TOAC is positioned just outside the binding motif, the binding constant remains nearly unaffected. Although the SH3 domain binds weakly and transiently to proline-rich peptides from FAK, the interaction is not very dynamic and the relative position of the spin-label to the protein is well-defined. It is concluded that TOAC can be used to generate reliable paramagnetic NMR restraints.

Mobility of TOAC spin-labelled peptides binding to the Src SH3 domain studied by paramagnetic NMR

International Nuclear Information System (INIS)

Lindfors, Hanna E.; Koning, Peter E. de; Wouter Drijfhout, Jan; Venezia, Brigida; Ubbink, Marcellus

2008-01-01

Paramagnetic relaxation enhancement provides a tool for studying the dynamics as well as the structure of macromolecular complexes. The application of side-chain coupled spin-labels is limited by the mobility of the free radical. The cyclic, rigid amino acid spin-label TOAC (2,2,6,6-Tetramethylpiperidine-1-oxyl-4-amino-4-carboxylic acid), which can be incorporated straightforwardly by peptide synthesis, provides an attractive alternative. In this study, TOAC was incorporated into a peptide derived from focal adhesion kinase (FAK), and the interaction of the peptide with the Src homology 3 (SH3) domain of Src kinase was studied, using paramagnetic NMR. Placing TOAC within the binding motif of the peptide has a considerable effect on the peptide-protein binding, lowering the affinity substantially. When the TOAC is positioned just outside the binding motif, the binding constant remains nearly unaffected. Although the SH3 domain binds weakly and transiently to proline-rich peptides from FAK, the interaction is not very dynamic and the relative position of the spin-label to the protein is well-defined. It is concluded that TOAC can be used to generate reliable paramagnetic NMR restraints

Combining biophysical methods to analyze the disulfide bond in SH2 domain of C-terminal Src kinase.

Science.gov (United States)

Liu, Dongsheng; Cowburn, David

2016-01-01

The Src Homology 2 (SH2) domain is a structurally conserved protein domain that typically binds to a phosphorylated tyrosine in a peptide motif from the target protein. The SH2 domain of C-terminal Src kinase (Csk) contains a single disulfide bond, which is unusual for most SH2 domains. Although the global motion of SH2 domain regulates Csk function, little is known about the relationship between the disulfide bond and binding of the ligand. In this study, we combined X-ray crystallography, solution NMR, and other biophysical methods to reveal the interaction network in Csk. Denaturation studies have shown that disulfide bond contributes significantly to the stability of SH2 domain, and crystal structures of the oxidized and C122S mutant showed minor conformational changes. We further investigated the binding of SH2 domain to a phosphorylated peptide from Csk-binding protein upon reduction and oxidation using both NMR and fluorescence approaches. This work employed NMR, X-ray cryptography, and other biophysical methods to study a disulfide bond in Csk SH2 domain. In addition, this work provides in-depth understanding of the structural dynamics of Csk SH2 domain.

The effect of temperature and time of extraction on the quality of Semi Refined Carrageenan (SRC

Directory of Open Access Journals (Sweden)

Heriyanto Heri

2018-01-01

Full Text Available Euchema cottonii is a good source of kappa-carrageenan and can be found cultivated in the Indonesia coastal areas in which one of them is in Banten Province. Carrageenans have many applications and are utilized in human food and pet-food industry. Carrageenans are also utilized in non-food industry such as pharmaceuticals, cosmetics, printing and textile formulations. Hence, the present study features on the cooking process cooking time and cooking temperature. The effects of these parameters on carrageenan quality such as gel viscosity and gel strength were studied. The process of extraction of carrageenan was conducted with variations temperature: 60, 70, and 80 °C and the variation of time: 1, 2, and 3 hours. Alkaline substance used was KOH with 8% concentration and the ratio of solvent to dry seaweed 8:1. From the present investigation, it was observed that SRC extraction reached the best condition at temperature 70 °C for 2 hours with the value of yield 30.20%, 5.90% moisture content, 18.34% ash content, sulfate content of 6.94%, viscosity of 190 cP, and the gel strength 714.45 g / cm2. The treatment of temperature and extraction time significantly affected the quality of the SRC yield parameter, viscosity and gel strength.

Optimal Couple Projections for Domain Adaptive Sparse Representation-based Classification.

Science.gov (United States)

Zhang, Guoqing; Sun, Huaijiang; Porikli, Fatih; Liu, Yazhou; Sun, Quansen

2017-08-29

In recent years, sparse representation based classification (SRC) is one of the most successful methods and has been shown impressive performance in various classification tasks. However, when the training data has a different distribution than the testing data, the learned sparse representation may not be optimal, and the performance of SRC will be degraded significantly. To address this problem, in this paper, we propose an optimal couple projections for domain-adaptive sparse representation-based classification (OCPD-SRC) method, in which the discriminative features of data in the two domains are simultaneously learned with the dictionary that can succinctly represent the training and testing data in the projected space. OCPD-SRC is designed based on the decision rule of SRC, with the objective to learn coupled projection matrices and a common discriminative dictionary such that the between-class sparse reconstruction residuals of data from both domains are maximized, and the within-class sparse reconstruction residuals of data are minimized in the projected low-dimensional space. Thus, the resulting representations can well fit SRC and simultaneously have a better discriminant ability. In addition, our method can be easily extended to multiple domains and can be kernelized to deal with the nonlinear structure of data. The optimal solution for the proposed method can be efficiently obtained following the alternative optimization method. Extensive experimental results on a series of benchmark databases show that our method is better or comparable to many state-of-the-art methods.

Combination Therapy with c-Met and Src Inhibitors Induces Caspase-Dependent Apoptosis of Merlin-Deficient Schwann Cells and Suppresses Growth of Schwannoma Cells.

Science.gov (United States)

Fuse, Marisa A; Plati, Stephani Klingeman; Burns, Sarah S; Dinh, Christine T; Bracho, Olena; Yan, Denise; Mittal, Rahul; Shen, Rulong; Soulakova, Julia N; Copik, Alicja J; Liu, Xue Zhong; Telischi, Fred F; Chang, Long-Sheng; Franco, Maria Clara; Fernandez-Valle, Cristina

2017-11-01

Neurofibromatosis type 2 (NF2) is a nervous system tumor disorder caused by inactivation of the merlin tumor suppressor encoded by the NF2 gene. Bilateral vestibular schwannomas are a diagnostic hallmark of NF2. Mainstream treatment options for NF2-associated tumors have been limited to surgery and radiotherapy; however, off-label uses of targeted molecular therapies are becoming increasingly common. Here, we investigated drugs targeting two kinases activated in NF2-associated schwannomas, c-Met and Src. We demonstrated that merlin-deficient mouse Schwann cells (MD-MSC) treated with the c-Met inhibitor, cabozantinib, or the Src kinase inhibitors, dasatinib and saracatinib, underwent a G 1 cell-cycle arrest. However, when MD-MSCs were treated with a combination of cabozantinib and saracatinib, they exhibited caspase-dependent apoptosis. The combination therapy also significantly reduced growth of MD-MSCs in an orthotopic allograft mouse model by greater than 80% of vehicle. Moreover, human vestibular schwannoma cells with NF2 mutations had a 40% decrease in cell viability when treated with cabozantinib and saracatinib together compared with the vehicle control. This study demonstrates that simultaneous inhibition of c-Met and Src signaling in MD-MSCs triggers apoptosis and reveals vulnerable pathways that could be exploited to develop NF2 therapies. Mol Cancer Ther; 16(11); 2387-98. ©2017 AACR . ©2017 American Association for Cancer Research.

Association of coatomer proteins with the beta-receptor for platelet-derived growth factor

DEFF Research Database (Denmark)

Hansen, Klaus; Rönnstrand, L; Rorsman, C

1997-01-01

The nonreceptor tyrosine kinase Src binds to and is activated by the beta-receptor for platelet-derived growth factor (PDGF). The interaction leads to Src phosphorylation of Tyr934 in the kinase domain of the receptor. In the course of the functional characterization of this phosphorylation, we...... of intracellular vesicle transport. In order to explore the functional significance of the interaction between alpha- and beta'-COP and the PDGF receptor, a receptor mutant was made in which the conserved histidine residue 928 was mutated to an alanine residue. The mutant receptor, which was unable to bind alpha...

Chemical and biological effects of heavy distillate recycle in the SRC-II process

Energy Technology Data Exchange (ETDEWEB)

Wilson, B.W.; Pelroy, R.A.; Anderson, R.P.; Freel, J.

1983-12-01

Recent work from the Merriam Laboratory continuous coal liquefaction units shows that heavy distillate from the SRC-II process can be recycled to extinction, and hence a distillate product boiling entirely below 310/sup 0/C (590/sup 0/F) (or other selected boiling points) is feasible. In these runs distillate yield was not reduced; gas make was unaffected; and hydrogen consumption was increased only slightly, in keeping with the generally higher hydrogen content of lighter end products. Total distillate yield (C/sub 5/-590/sup 0/F) was 56 wt %, MAF coal in runs with subbituminous coal from the Amax Belle Ayr mine. Product endpoint is well below 371/sup 0/C (700/sup 0/F), the temperature above which coal distillates appear to become genotoxic; and the product was shown to be free of mutagenic activity in the Ames test. Chemical analyses showed both the < 270/sup 0/C (< 518/sup 0/F) and the < 310/sup 0/C (< 590/sup 0/F) distillates to be essentially devoid of several reference polycyclic compounds known to be carcinogenic in laboratory animals. Tests for tumorigenic or carcinogenic activity were not carried out on these materials. However, a comparison of chemical data from the Merriam heavy distillate samples with data on the other SRC-II distillates where carcinogenesis or tumorigenesis data is available leads to the expectation that < 371/sup 0/C (< 700/sup 0/F) materials from the Merriam Laboratory will have greatly reduced tumorigenic and carcinogenic activity in skin painting tests. Other studies suggest the product should be more readily upgraded than full-range (C/sub 5/-900/sup 0/F) distillate.

Suppressed invasive and migratory behaviors of SW1353 chondrosarcoma cells through the regulation of Src, Rac1 GTPase, and MMP13.

Science.gov (United States)

Xu, Wenxiao; Wan, Qiaoqiao; Na, Sungsoo; Yokota, Hiroki; Yan, Jing-Long; Hamamura, Kazunori

2015-12-01

Chondrosarcoma is the second frequent type of primary bone cancer. In response to stress to the endoplasmic reticulum, activation of eIF2α-mediated signaling is reported to induce apoptosis. However, its effects on invasive and migratory behaviors of chondrosarcoma have not been understood. Focusing on potential roles of Src kinase, Rac1 GTPase, and MMP13, we investigated eIF2α-driven regulation of SW1353 chondrosarcoma cells. In particular, we employed two chemical agents (salubrinal, Sal; and guanabenz, Gu) that elevate the level of eIF2α phosphorylation. The result revealed that both Sal and Gu reduced invasion and motility of SW1353 chondrosarcoma cells in a dose dependent manner. Live imaging using a fluorescent resonance energy transfer (FRET) technique showed that Sal and Gu downregulated activities of Src kinase as well as Rac1 GTPase in an eIF2α dependent manner. RNA interference experiments supported an eIF2α-mediated regulatory network in the inhibitory role of Sal and Gu. Partial silencing of MMP13 also suppressed malignant phenotypes of SW1353 chondrosarcoma cells. However, MMP13 was not regulated via eIF2α since administration of Sal but not Gu reduced expression of MMP13. In summary, we demonstrate that eIF2α dependent and independent pathways regulate invasion and motility of SW1353 chondrosarcoma cells, and inactivation of Src, Rac1, and MMP13 by Sal could provide a potential adjuvant therapy for combating metastatic chondrosarcoma cells. Copyright © 2015 Elsevier Inc. All rights reserved.

Spontaneous regression of residual low-grade cerebellar pilocytic astrocytomas in children

International Nuclear Information System (INIS)

Gunny, Roxana S.; Saunders, Dawn E.; Hayward, Richard D.; Phipps, Kim P.; Harding, Brian N.

2005-01-01

Cerebellar low-grade astrocytomas (CLGAs) of childhood are benign tumours and are usually curable by surgical resection alone or combined with adjuvant radiotherapy. To undertake a retrospective study of our children with CLGA to determine the optimum schedule for surveillance imaging following initial surgery. In this report we describe the phenomenon of spontaneous regression of residual tumour and discuss its prognostic significance regarding future imaging. A retrospective review was conducted of children treated for histologically proven CLGA at Great Ormond Street Hospital from 1988 to 1998. Of 83 children with CLGA identified, 13 (15.7%) had incomplete resections. Two children with large residual tumours associated with persistent symptoms underwent additional treatment. Eleven children were followed by surveillance imaging alone for a mean of 6.83 years (range 2-13.25 years). Spontaneous tumour regression was seen in 5 (45.5%) of the 11 children. There were no differences in age, gender, symptomatology, histological grade or Ki-67 fractions between those with spontaneous tumour regression and those with progression. There was a non-significant trend that larger volume residual tumours progressed. Residual tumour followed by surveillance imaging may either regress or progress. For children with residual disease we recommend surveillance imaging every 6 months for the first 2 years, every year for years 3, 4 and 5, then every second year if residual tumour is still present 5 years after initial surgery. This would detect not only progressive or recurrent disease, but also spontaneous regression which can occur later than disease progression. (orig.)

Climate Data Provenance Tracking for Just-In-Time Computation

Science.gov (United States)

Fries, S.; Nadeau, D.; Doutriaux, C.; Williams, D. N.

2016-12-01

The "Climate Data Management System" (CDMS) was created in 1996 as part of the Climate Data Analysis Tools suite of software. It provides a simple interface into a wide variety of climate data formats, and creates NetCDF CF-Compliant files. It leverages the NumPy framework for high performance computation, and is an all-in-one IO and computation package. CDMS has been extended to track manipulations of data, and trace that data all the way to the original raw data. This extension tracks provenance about data, and enables just-in-time (JIT) computation. The provenance for each variable is packaged as part of the variable's metadata, and can be used to validate data processing and computations (by repeating the analysis on the original data). It also allows for an alternate solution for sharing analyzed data; if the bandwidth for a transfer is prohibitively expensive, the provenance serialization can be passed in a much more compact format and the analysis rerun on the input data. Data provenance tracking in CDMS enables far-reaching and impactful functionalities, permitting implementation of many analytical paradigms.

The Emerging and Diverse Roles of Src-Like Adaptor Proteins in Health and Disease

Directory of Open Access Journals (Sweden)

Nikolett Marton

2015-01-01

Full Text Available Although Src-like adaptor proteins (SLAP-1 and SLAP-2 were mainly studied in lymphocytes, where they act as negative regulators and provide fine control of receptor signaling, recently, several other functions of these proteins were discovered. In addition to the well-characterized immunoregulatory functions, SLAP proteins appear to have an essential role in the pathogenesis of type I hypersensitivity, osteoporosis, and numerous malignant diseases. Both adaptor proteins are expressed in a wide variety of tissues, where they have mostly inhibitory effects on multiple intracellular signaling pathways. In this review, we summarize the diverse effects of SLAP proteins.

Vest-over-Pant Method for Closure of Residual Cavity of Liver Hydatid Cyst

Directory of Open Access Journals (Sweden)

Saba Behdad

2012-01-01

Full Text Available Objective. Although several therapeutic strategies have proven to be effective for hydatid cyst of liver, but surgery is still the most common therapy despite its morbidity and mortality. Furthermore, a variety of technique has been recommended for managing the residual cavity after cystectomy. We report here a new technical method for the reconstruction of hydatid cyst residual cavity with using overlapping flaps of liver edges (Vest over Pant. Methods. In this technique after removing the cyst, the edges of one side of cyst cavity were sutured to the base of the cavity using three to four mattress sutures, and edges of other side of liver was overlapped on the dorsal part of previous layer using four to five mattress sutures. Therefore residual cavity dead space was obliterated with two surfaces of cavity. Results. Fifty males were treated by our method. The average cyst volume was 423±110 mL. There was no intraabdominal sepsis, bile leakage, or hepatic necrosis. In follow-up ultrasound study, residual cavities were disappeared one month after operation. Conclusion. Overlapping flaps of liver edges (Vest over Pant provides easy, safe closure of cyst with preservation of the liver anatomy.

Sudetic larch in Germany - Results of provenance and progeny research

Energy Technology Data Exchange (ETDEWEB)

Weisgerber, H. [Forest Centre for Management Planning, Research and Ecology, Hann Muenden (Germany)

1995-12-31

There are only a few older sources of Sudetic larch in Germany. They distinguish themselves by outstanding growth and low susceptibility to canker. This impression was confirmed by results of provenance research. The Sudetic larch tested in comparison with numerous other provenances proved to be fast-growing, site-tolerant, to a large extent insusceptible to canker, with straight but also slightly to moderately curved stems. The Sudetic provenances behave remarkably uniformly as regards these characteristics. In addition to provenance research investigations have been going on for a long time in Germany into individual differences within the Sudetic larch populations. A report is given on the results of progeny tests from free and controlled pollination, using the example of a seed orchard consisting of 54 clones. We point also to possibilities for improving stem quality by selection steps. The results of provenance and progeny research on Sudetic larch are in the meantime being put to use to a large extent in practical forestry. The forest administrations of various federal lands recommend the use of reproductive material of Sudetic origin and from seed orchards. 20 refs, 3 figs

Sudetic larch in Germany - Results of provenance and progeny research

Energy Technology Data Exchange (ETDEWEB)

Weisgerber, H [Forest Centre for Management Planning, Research and Ecology, Hann Muenden (Germany)

1996-12-31

There are only a few older sources of Sudetic larch in Germany. They distinguish themselves by outstanding growth and low susceptibility to canker. This impression was confirmed by results of provenance research. The Sudetic larch tested in comparison with numerous other provenances proved to be fast-growing, site-tolerant, to a large extent insusceptible to canker, with straight but also slightly to moderately curved stems. The Sudetic provenances behave remarkably uniformly as regards these characteristics. In addition to provenance research investigations have been going on for a long time in Germany into individual differences within the Sudetic larch populations. A report is given on the results of progeny tests from free and controlled pollination, using the example of a seed orchard consisting of 54 clones. We point also to possibilities for improving stem quality by selection steps. The results of provenance and progeny research on Sudetic larch are in the meantime being put to use to a large extent in practical forestry. The forest administrations of various federal lands recommend the use of reproductive material of Sudetic origin and from seed orchards. 20 refs, 3 figs

Requirement of ERα and basal activities of EGFR and Src kinase in Cd-induced activation of MAPK/ERK pathway in human breast cancer MCF-7 cells

Energy Technology Data Exchange (ETDEWEB)

Song, Xiulong, E-mail: songxiulong@hotmail.com; Wei, Zhengxi; Shaikh, Zahir A., E-mail: zshaikh@uri.edu

2015-08-15

Cadmium (Cd) is a common environmental toxicant and an established carcinogen. Epidemiological studies implicate Cd with human breast cancer. Low micromolar concentrations of Cd promote proliferation of human breast cancer cells in vitro. The growth promotion of breast cancer cells is associated with the activation of MAPK/ERK pathway. This study explores the mechanism of Cd-induced activation of MAPK/ERK pathway. Specifically, the role of cell surface receptors ERα, EGFR, and Src kinase was evaluated in human breast cancer MCF-7 cells treated with 1–3 μM Cd. The activation of ERK was studied using a serum response element (SRE) luciferase reporter assay. Receptor phosphorylation was detected by Western blot analyses. Cd treatment increased both the SRE reporter activity and ERK1/2 phosphorylation in a concentration-dependent manner. Cd treatment had no effect on reactive oxygen species (ROS) generation. Also, blocking the entry of Cd into the cells with manganese did not diminish Cd-induced activation of MAPK/ERK. These results suggest that the effect of Cd was likely not caused by intracellular ROS generation, but through interaction with the membrane receptors. While Cd did not appear to activate either EGFR or Src kinase, their inhibition completely blocked the Cd-induced activation of ERK as well as cell proliferation. Similarly, silencing ERα with siRNA or use of ERα antagonist blocked the effects of Cd. Based on these results, it is concluded that not only ERα, but also basal activities of EGFR and Src kinase are essential for Cd-induced signal transduction and activation of MAPK/ERK pathway for breast cancer cell proliferation. - Highlights: • Low micromolar concentrations of Cd rapidly activate ERK1/2 in MCF-7 cells. • Signal transduction and resulting cell proliferation require EGFR, ERα, and Src. • These findings implicate Cd in promotion of breast cancer.

Structural and Biochemical Characterization of SrcA, a Multi-cargo Type III Secretion Chaperone in Salmonella Required for Pathogenic Association with a Host

Energy Technology Data Exchange (ETDEWEB)

Cooper, C.; Zhang, K; Andres, S; Fnag, Y; Kaniuk, N; Hannemann, M; Brumell, J; Foster, L; Junop, M; Coombes, B

2010-01-01

Many Gram-negative bacteria colonize and exploit host niches using a protein apparatus called a type III secretion system (T3SS) that translocates bacterial effector proteins into host cells where their functions are essential for pathogenesis. A suite of T3SS-associated chaperone proteins bind cargo in the bacterial cytosol, establishing protein interaction networks needed for effector translocation into host cells. In Salmonella enterica serovar Typhimurium, a T3SS encoded in a large genomic island (SPI-2) is required for intracellular infection, but the chaperone complement required for effector translocation by this system is not known. Using a reverse genetics approach, we identified a multi-cargo secretion chaperone that is functionally integrated with the SPI-2-encoded T3SS and required for systemic infection in mice. Crystallographic analysis of SrcA at a resolution of 2.5 {angstrom} revealed a dimer similar to the CesT chaperone from enteropathogenic E. coli but lacking a 17-amino acid extension at the carboxyl terminus. Further biochemical and quantitative proteomics data revealed three protein interactions with SrcA, including two effector cargos (SseL and PipB2) and the type III-associated ATPase, SsaN, that increases the efficiency of effector translocation. Using competitive infections in mice we show that SrcA increases bacterial fitness during host infection, highlighting the in vivo importance of effector chaperones for the SPI-2 T3SS.

Structural and biochemical characterization of SrcA, a multi-cargo type III secretion chaperone in Salmonella required for pathogenic association with a host.

Directory of Open Access Journals (Sweden)

Colin A Cooper

2010-02-01

Full Text Available Many Gram-negative bacteria colonize and exploit host niches using a protein apparatus called a type III secretion system (T3SS that translocates bacterial effector proteins into host cells where their functions are essential for pathogenesis. A suite of T3SS-associated chaperone proteins bind cargo in the bacterial cytosol, establishing protein interaction networks needed for effector translocation into host cells. In Salmonella enterica serovar Typhimurium, a T3SS encoded in a large genomic island (SPI-2 is required for intracellular infection, but the chaperone complement required for effector translocation by this system is not known. Using a reverse genetics approach, we identified a multi-cargo secretion chaperone that is functionally integrated with the SPI-2-encoded T3SS and required for systemic infection in mice. Crystallographic analysis of SrcA at a resolution of 2.5 A revealed a dimer similar to the CesT chaperone from enteropathogenic E. coli but lacking a 17-amino acid extension at the carboxyl terminus. Further biochemical and quantitative proteomics data revealed three protein interactions with SrcA, including two effector cargos (SseL and PipB2 and the type III-associated ATPase, SsaN, that increases the efficiency of effector translocation. Using competitive infections in mice we show that SrcA increases bacterial fitness during host infection, highlighting the in vivo importance of effector chaperones for the SPI-2 T3SS.

Enhanced Prediction of Src Homology 2 (SH2) Domain Binding Potentials Using a Fluorescence Polarization-derived c-Met, c-Kit, ErbB, and Androgen Receptor Interactome*

Science.gov (United States)

Leung, Kin K.; Hause, Ronald J.; Barkinge, John L.; Ciaccio, Mark F.; Chuu, Chih-Pin; Jones, Richard B.

2014-01-01

Many human diseases are associated with aberrant regulation of phosphoprotein signaling networks. Src homology 2 (SH2) domains represent the major class of protein domains in metazoans that interact with proteins phosphorylated on the amino acid residue tyrosine. Although current SH2 domain prediction algorithms perform well at predicting the sequences of phosphorylated peptides that are likely to result in the highest possible interaction affinity in the context of random peptide library screens, these algorithms do poorly at predicting the interaction potential of SH2 domains with physiologically derived protein sequences. We employed a high throughput interaction assay system to empirically determine the affinity between 93 human SH2 domains and phosphopeptides abstracted from several receptor tyrosine kinases and signaling proteins. The resulting interaction experiments revealed over 1000 novel peptide-protein interactions and provided a glimpse into the common and specific interaction potentials of c-Met, c-Kit, GAB1, and the human androgen receptor. We used these data to build a permutation-based logistic regression classifier that performed considerably better than existing algorithms for predicting the interaction potential of several SH2 domains. PMID:24728074

Seamless Provenance Representation and Use in Collaborative Science Scenarios

Science.gov (United States)

Missier, P.; Ludaescher, B.; Bowers, S.; Altintas, I.; Anand, M. K.; Dey, S.; Sarkar, A.; Shrestha, B.; Goble, C.

2010-12-01

The notion of sharing scientific data has only recently begun to gain ground in science, where data is still considered a private asset. There is growing evidence, however, that the benefits of scientific collaboration through early data sharing during the course of a science project may outgrow the risk of losing exclusive ownership of the data. As exemplar success stories are making the headlines[1], principles of effective information sharing have become the subject of e-science research. In particular, any piece of published data should be self-describing, to the extent necessary for consumers to determine its suitability for reuse in their own projects. This is accomplished by associating a body of formally specified and machine-processable metadata to the data. When data is produced and reused by independent groups, however, metadata interoperability issues emerge. This is the case for provenance, a form of metadata that describes the history of a data product, Y. Provenance is typically expressed as a graph-structured set of dependencies that account for the sequence of computational or interactive steps that led to Y, often starting from some primary, observational data. Traversing dependency graphs is one of the mechanisms used to answer questions on data reliability. In the context of the NSF DataONE project[2], we have been studying issues of provenance interoperability in scientific collaboration scenarios. Consider a first scientist, Alice, who publishes a data product X along with its provenance, and a second scientist who further transforms X into a new product Y, also along with its provenance. A third scientist, who is interested in Y, expects to be able to trace Y's history up to the inputs used by Alice. This is only possible, however, if provenance accumulates into a single, uniform graph that can be seamlessly traversed. This becomes problematic when provenance is captured using different tools and computational models (i.e. workflow systems

Chaperone-like activity of β-casein and its effect on residual in vitro activity of horseradish peroxidase

DEFF Research Database (Denmark)

Sulewska, Anna Maria; Olsen, Karsten; Sørensen, Jens Christian

2014-01-01

, as similar experiment with bovine serum albumin resulted in residual activity of horseradish peroxidase that was significantly lower than without any addition. The effect of β-casein on HRP disappears when pH is below the isoelectric point of β-casein. It was also proven by light scattering studies that β...... proteins. Incubating HRP (0.1 mg mL-1) for 10 min at 72 °C resulted in residual activity of 59 ± 5%, while addition of 1 mg mL-1 β-casein resulted in increase in residual activity up to 85 ± 1%. Increased residual activity is not merely attributed to an effect of higher total protein concentration......-casein interacts with horseradish peroxidase when the temperature was increased from 25 to 70 °C whereas interactions seem to cease when temperature was lowered back to 25 °C. This study highlights how specific proteins can influence enzyme activity, which is of potential importance for various industries...

Complexes of gamma-tubulin with nonreceptor protein tyrosine kinases Src and Fyn in differentiating P19 embryonal carcinoma cells

Czech Academy of Sciences Publication Activity Database

Kukharskyy, Vitaliy; Sulimenko, Vadym; Macůrek, Libor; Sulimenko, Tetyana; Dráberová, Eduarda; Dráber, Pavel

2004-01-01

Roč. 298, - (2004), s. 218-228 ISSN 0014-4827 R&D Projects: GA AV ČR IAA5052004; GA ČR GA304/00/0553; GA ČR GA304/04/1273; GA MŠk LN00A026 Keywords : gamma-tubulin * P19 cells * Fyn and Src kinase Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 4.007, year: 2004

Determination of marble provenance: limits of isotopic analysis

International Nuclear Information System (INIS)

Germann, K.; Holzmann, G.; Winkler, F.J.

1980-01-01

Provenance determination of Thessalian stelae marbles using the C/O isotopic analysis proved to be misleading, as the isotopic composition even in very small quarrying areas is heterogeneous and isotopic coincidence of marbles from very distant sources occurs. Therefore additional geological features must be taken into consideration and preference should be given to combinations of both petrographical and geochemical properties. Geological field work to establish the range of possible marble sources and the variability within these sources is one of the prerequisites of provenance studies. (author)

Fagus sylvatica L. provenances maintain different leaf metabolic profiles and functional response

Science.gov (United States)

Aranda, Ismael; Sánchez-Gómez, David; de Miguel, Marina; Mancha, Jose Antonio; Guevara, María Angeles; Cadahía, Estrella; Fernández de Simón, María Brígida

2017-07-01

Most temperate forest tree species will suffer important environmental changes as result of the climate change. Adaptiveness to local conditions could change at different sites in the future. In this context, the study of intra-specific variability is important to clarify the singularity of different local populations. Phenotypic differentiation between three beech provenances covering a wide latitudinal range (Spain/ES, Germany/DE and Sweden/SE), was studied in a greenhouse experiment. Non-target leaf metabolite profiles and ecophysiological response was analyzed in well-watered and water stressed seedlings. There was a provenance-specific pattern in the relative concentrations of some leaf metabolites regardless watering treatment. The DE and SE from the center and north of the distribution area of the species showed a clear differentiation from the ES provenance in the relative concentration of some metabolites. Thus the ES provenance from the south maintained larger relative concentration of some organic and amino acids (e.g. fumaric and succinic acids or valine and isoleucine), and in some secondary metabolites (e.g. kaempferol, caffeic and ferulic acids). The ecophysiological response to mild water stress was similar among the three provenances as a consequence of the moderate water stress applied to seedlings, although leaf N isotope composition (δ15N) and leaf C:N ratio were higher and lower respectively in DE than in the other two provenances. This would suggest potential differences in the capacity to uptake and post-process nitrogen according to provenance. An important focus of the study was to address for the first time inter-provenance leaf metabolic diversity in beech from a non-targeted metabolic profiling approach that allowed differentiation of the three studied provenances.

A HIF-regulated VHL-PTP1B-Src signaling axis identifies a therapeutic target in Renal Cell Carcinoma

OpenAIRE

Suwaki, Natsuko; Vanhecke, Elsa; Atkins, Katelyn M.; Graf, Manuela; Swabey, Katherine; Huang, Paul; Schraml, Peter; Moch, Holger; Cassidy, Amy; Brewer, Daniel; Al-Lazikani, Bissan; Workman, Paul; De-Bono, Johann; Kaye, Stan B.; Larkin, James

2011-01-01

Metastatic renal cell carcinoma (RCC) is a molecularly heterogeneous disease that is intrinsically resistant to chemotherapy and radiotherapy. While VEGF and mTOR targeted therapies have shown clinical activity, their effects are variable and short-lived, underscoring the need for improved treatment strategies for RCC. Here, we used quantitative phosphoproteomics and immunohistochemical profiling of 346 RCC specimens to determine that Src kinase signaling is elevated in RCC cells that retain ...

Pharmacists' Perception of the Sale of Non-Clinically Proven Health ...

African Journals Online (AJOL)

Pharmacists' Perception of the Sale of Non-Clinically Proven Health Supplements in Penang, Malaysia. ... A total of 10.7 % respondents indicated that the sale of non-clinically proven products result in high profit. Only 25.0 % of the pharmacists ... Keywords: Perception, Health promotion, Urban poor, Health supplements.

MT1-MMP promotes cell growth and ERK activation through c-Src and paxillin in three-dimensional collagen matrix

International Nuclear Information System (INIS)

Takino, Takahisa; Tsuge, Hisashi; Ozawa, Terumasa; Sato, Hiroshi

2010-01-01

Membrane-type 1 matrix metalloproteinase (MT1-MMP) is essential for tumor invasion and growth. We show here that MT1-MMP induces extracellular signal-regulated kinase (ERK) activation in cancer cells cultured in collagen gel, which is indispensable for their proliferation. Inhibition of MT1-MMP by MMP inhibitor or small interfering RNA suppressed activation of focal adhesion kinase (FAK) and ERK in MT1-MMP-expressing cancer cells, which resulted in up-regulation of p21 WAF1 and suppression of cell growth in collagen gel. Cell proliferation was also abrogated by the inhibitor against ERK pathway without affecting FAK phosphorylation. MT1-MMP and integrin α v β 3 were shown to be involved in c-Src activation, which induced FAK and ERK activation in collagen gel. These MT1-MMP-mediated signal transductions were paxillin dependent, as knockdown of paxillin reduced cell growth and ERK activation, and co-expression of MT1-MMP with paxillin induced ERK activation. The results suggest that MT1-MMP contributes to proliferation of cancer cells in the extracellular matrix by activating ERK through c-Src and paxillin.

Using Nd-Sr isotopes and rare earth elements to study sediment provenance of the modern radial sand ridges in the southwestern Yellow Sea

International Nuclear Information System (INIS)

Rao, Wenbo; Mao, Changping; Wang, Yigang; Huang, Huiming; Ji, Junfeng

2017-01-01

The radial sand ridges (RSRs) in the southwestern Yellow Sea off the Jiangsu Coast, East China have been intensively studied at least since 1975. Despite decades of studies, the provenance of the RSR sediments remains uncertain. In this study, the Nd-Sr isotopic and REE geochemical compositions of residual sediments (i.e., the acid-insoluble fractions) were investigated to determine the provenance of the RSR sediments. The Nd isotopic composition, PAAS-normalized REE patterns and characteristic parameters (e.g., Sm/Nd, (La/Sm)_N, (Gd/Yb)_N) were merely associated with source rocks but not with particle sorting while the Sr isotopic composition and REE contents of residual sediments were affected by particle sorting in addition to source rocks. The onshore RSR sediments originated mainly from mixing of the fine-grained sediments from various parts of the offshore RSR in terms of REE geochemical and isotopic analyses. Isotopic and REE geochemical comparison further reveals that the RSRs off the Jiangsu Coast were fed chiefly by the dispersal of surface sediments from the Yangtze River Mouth. Surface sediments from the Yangtze River Mouth were directly dispersed to the RSRs along the Jiangsu Coast and significantly affected the seaward part of the offshore RSR and the old Yellow River Delta area by a northward branch of the Changjiang Diluted Freshwater Plume. Only minor quantities of surface sediments from the modern Yellow River Mouth were introduced into the RSRs by the Jiangsu Coastal Current and mainly contaminated the landward part of the offshore RSR area. Our findings highlighted the potential of the Nd isotopes with REE geochemistry to trace the provenance of coastal sediments. - Highlights: • Nd isotopic and REE geochemical compositions of coastal and estuary sediments are mainly associated with source rocks. • Onshore RSR sediments originate from mixing of fine-grained sediments from various parts of the offshore RSR. • The RSRs off the Jiangsu Coast

Model-based Abstraction of Data Provenance

NARCIS (Netherlands)

Probst, Christian W.; Hansen, René Rydhof

Identifying provenance of data provides insights to the origin of data and intermediate results, and has recently gained increased interest due to data-centric applications. In this work we extend a data-centric system view with actors handling the data and policies restricting actions. This

Residual nilpotence and residual solubility of groups

International Nuclear Information System (INIS)

Mikhailov, R V

2005-01-01

The properties of the residual nilpotence and the residual solubility of groups are studied. The main objects under investigation are the class of residually nilpotent groups such that each central extension of these groups is also residually nilpotent and the class of residually soluble groups such that each Abelian extension of these groups is residually soluble. Various examples of groups not belonging to these classes are constructed by homological methods and methods of the theory of modules over group rings. Several applications of the theory under consideration are presented and problems concerning the residual nilpotence of one-relator groups are considered.

The Src family kinase inhibitor dasatinib delays pain-related behaviour and conserves bone in a rat model of cancer-induced bone pain

DEFF Research Database (Denmark)

Appel, Camilla Kristine; Gallego-Pedersen, Simone; Andersen, Line

2017-01-01

-induced bone pain, including cancer growth, osteoclastic bone degradation and nociceptive signalling. Here we investigate the role of dasatinib, an oral Src kinase family and Bcr-Abl tyrosine kinase inhibitor, in an animal model of cancer-induced bone pain. Daily administration of dasatinib (15 mg/kg, p...

Growth dynamics variation of different larch provenances under the mountain conditions in Poland

Energy Technology Data Exchange (ETDEWEB)

Kulej, M. [Univ. of Agriculture, Cracow (Poland). Section of Seed Production and Selection

1995-12-31

The results of 25-year investigations based on measurements and statistical analysis concerning the growth dynamics variation of larch provenances from the entire area of Poland are reported in this paper. This is the first larch provenance experiment in Poland under mountain conditions. The results obtained showed a significant variability among the provenances tested as regards the basic growth characters (height, d.b.h., growth index) at the age of 5, 8, 11, 15, 20 and 25 years. The larch from Klodzko and Proszkow turned out to be the best in respect of growth during the entire 25-years period. Decidedly bad were provenances from Marcule, Grojec, Rawa mazowiecka and Kroscienko. We cannot forecast the future growth of larch when trees are 5-years old since such prognosis may carry an error. However, on the basis of the results obtained it may be concluded that when trees are about 8 years old the stabilization of the position of individual provenances as regards growth takes place. The height growth curves for the individual provenances during the 25-years period (with exception of the provenance from Marcule) fall within the interval {+-} 0,5S from the compensated curve for the entire population studied. All larch provenances in the experiment had reached the height growth culmination. A greatest differentiation in respect of this character occurred in case of the provenances from Sudetes. 27 refs, 4 figs, 8 tabs

Growth dynamics variation of different larch provenances under the mountain conditions in Poland

Energy Technology Data Exchange (ETDEWEB)

Kulej, M [Univ. of Agriculture, Cracow (Poland). Section of Seed Production and Selection

1996-12-31

The results of 25-year investigations based on measurements and statistical analysis concerning the growth dynamics variation of larch provenances from the entire area of Poland are reported in this paper. This is the first larch provenance experiment in Poland under mountain conditions. The results obtained showed a significant variability among the provenances tested as regards the basic growth characters (height, d.b.h., growth index) at the age of 5, 8, 11, 15, 20 and 25 years. The larch from Klodzko and Proszkow turned out to be the best in respect of growth during the entire 25-years period. Decidedly bad were provenances from Marcule, Grojec, Rawa mazowiecka and Kroscienko. We cannot forecast the future growth of larch when trees are 5-years old since such prognosis may carry an error. However, on the basis of the results obtained it may be concluded that when trees are about 8 years old the stabilization of the position of individual provenances as regards growth takes place. The height growth curves for the individual provenances during the 25-years period (with exception of the provenance from Marcule) fall within the interval {+-} 0,5S from the compensated curve for the entire population studied. All larch provenances in the experiment had reached the height growth culmination. A greatest differentiation in respect of this character occurred in case of the provenances from Sudetes. 27 refs, 4 figs, 8 tabs

Energy generation for sustainable development with innovation technology and utilization of biomass residue; Geracao de energia para o desenvolvimento rural sustentavel com inovacao tecnologica de aproveitamento de biomassa residual

Energy Technology Data Exchange (ETDEWEB)

Fernandes, Maria Roseane de Pontes; Lopes, Carlos Eduardo Bezerra; Costa Neto, Manoel Bezerra da; Selvam, P.V. Pannir [Universidade Federal do Rio Grande do Norte (UFRN), Natal, RN (Brazil)

2004-07-01

In the present work, the introduction of alternative energy of biogas in agricultural communities for the sustainable development was studied through exploitation of residual biomass and also getting as by-product the biological fertilizer. A fast composting of the domestic residue with the organic was made possible where part of this residue after processing was taken together with effluent to the biodigestor. The bibliographical research on the processes of generation of biogas, about composting and the equipment for processing had been carried through. The projects Engineering with the use of computational tools had been developed with the Software Super Pro 4,9 Design and ORC GPEC 2004 by our research group. Five case studies had been elaborated, where different scenes related with our innovation, that uses of the residue for the composting together with domestic effluent for digestion. Several economic parameters were obtained and our work proved the viability about the use of biogas for drying of the fruits banana. A economic feasibility study was carried where it was proven that the project with the innovation of the use of residues from the fruits possesses more advantages than the conventional system of drying using electric energy. Considering the viability of this process and the use solar energy, it is intended to apply this technology in rural agricultural communities providing them an energy source of low cost in substitution of the conventional energy. (author)

On the importance of a funneled energy landscape for the assembly and regulation of multidomain Src tyrosine kinases

OpenAIRE

Faraldo-Gómez, José D.; Roux, Benoît

2007-01-01

Regulation of signaling pathways in the cell often involves multidomain allosteric enzymes that are able to adopt alternate active or inactive conformations in response to specific stimuli. It is therefore of great interest to elucidate the energetic and structural determinants that govern the conformational plasticity of these proteins. In this study, free-energy computations have been used to address this fundamental question, focusing on one important family of signaling enzymes, the Src t...

PI3K/Akt Activated by GPR30 and Src Regulates 17β-Estradiol-Induced Cultured Immature Boar Sertoli Cells Proliferation.

Science.gov (United States)

Yang, Wei-Rong; Zhu, Feng-Wei; Zhang, Jiao-Jiao; Wang, Yi; Zhang, Jia-Hua; Lu, Cheng; Wang, Xian-Zhong

2016-05-24

Sertoli cell (SC) is a key element in the process of spermatogenesis. Accumulating research show that estrogen plays an important role in regulating boar SC proliferation. However, it is unclear whether phosphatidylinositol-4,5-bisphosphate 3-kinase/protein kinase B (PI3K/Akt) is involved in this process. In the present study, the role of PI3K/Akt on the 17β-estradiol-induced piglet SC proliferation was explored. In addition, we also explained the roles of G-protein-coupled estrogen receptor (GPR30) and Sarcoma protein (Src) in this process. Our study demonstrated that, 17β-estradiol induced activation of PI3K in a time-dependent manner. Both G-15 (an antagonist of GPR30, 0.1 μmol/L) and PP2 (an inhibitor of Src, 2.0 μmol/L) inhibited 17β-estradiol-induced activation of PI3K, reduced SC proliferation, and decreased messenger RNA (mRNA) and protein expression of S-phase kinase-associated protein 2 (Skp2). We also found that 17β-estradiol induced activation of Akt in a time-dependent manner. Both LY294002 (an inhibitor of PI3K) and 10-DEBC (an inhibitor of Akt) significantly reduced 17β-estradiol-induced SC proliferation and reduced mRNA and protein expression of Skp2. In addition, LY294002 inhibited 17β-estradiol-induced activation of Akt. The results indicated that 17β-estradiol regulates SC proliferation by activating PI3K/Akt. Both GPR30 and Src are involved in 17β-estradiol-induced phosphorylation of PI3K/Akt. Activation of PI3K/Akt enhances the expression of Skp2, which promotes SC proliferation. © The Author(s) 2016.

V-src oncogene-specific carboxy-terminal peptide is immunoprotective against Rous sarcoma growth in chickens with MHC class I allele B-F12

Czech Academy of Sciences Publication Activity Database

Hofmann, A.; Plachý, Jiří; Hunt, L.; Kaufman, J.; Hála, K.

2003-01-01

Roč. 2003, č. 21 (2003), s. 4694-4699 ISSN 0264-410X Institutional research plan: CEZ:AV0Z5052915 Keywords : Rous sarcoma * v-src peptide * chicken MHC class I allele Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.007, year: 2003

Presence of an SH2 domain in the actin-binding protein tensin.

Science.gov (United States)

Davis, S; Lu, M L; Lo, S H; Lin, S; Butler, J A; Druker, B J; Roberts, T M; An, Q; Chen, L B

1991-05-03

The molecular cloning of the complementary DNA coding for a 90-kilodalton fragment of tensin, an actin-binding component of focal contacts and other submembraneous cytoskeletal structures, is reported. The derived amino acid sequence revealed the presence of a Src homology 2 (SH2) domain. This domain is shared by a number of signal transduction proteins including nonreceptor tyrosine kinases such as Abl, Fps, Src, and Src family members, the transforming protein Crk, phospholipase C-gamma 1, PI-3 (phosphatidylinositol) kinase, and guanosine triphosphatase-activating protein (GAP). Like the SH2 domain found in Src, Crk, and Abl, the SH2 domain of tensin bound specifically to a number of phosphotyrosine-containing proteins from v-src-transformed cells. Tensin was also found to be phosphorylated on tyrosine residues. These findings suggest that by possessing both actin-binding and phosphotyrosine-binding activities and being itself a target for tyrosine kinases, tensin may link signal transduction pathways with the cytoskeleton.

Automated Generation of Technical Documentation and Provenance for Reproducible Research

Science.gov (United States)

Jolly, B.; Medyckyj-Scott, D.; Spiekermann, R.; Ausseil, A. G.

2017-12-01

Data provenance and detailed technical documentation are essential components of high-quality reproducible research, however are often only partially addressed during a research project. Recording and maintaining this information during the course of a project can be a difficult task to get right as it is a time consuming and often boring process for the researchers involved. As a result, provenance records and technical documentation provided alongside research results can be incomplete or may not be completely consistent with the actual processes followed. While providing access to the data and code used by the original researchers goes some way toward enabling reproducibility, this does not count as, or replace, data provenance. Additionally, this can be a poor substitute for good technical documentation and is often more difficult for a third-party to understand - particularly if they do not understand the programming language(s) used. We present and discuss a tool built from the ground up for the production of well-documented and reproducible spatial datasets that are created by applying a series of classification rules to a number of input layers. The internal model of the classification rules required by the tool to process the input data is exploited to also produce technical documentation and provenance records with minimal additional user input. Available provenance records that accompany input datasets are incorporated into those that describe the current process. As a result, each time a new iteration of the analysis is performed the documentation and provenance records are re-generated to provide an accurate description of the exact process followed. The generic nature of this tool, and the lessons learned during its creation, have wider application to other fields where the production of derivative datasets must be done in an open, defensible, and reproducible way.

Lowering the Barrier to Reproducible Research by Publishing Provenance from Common Analytical Tools

Science.gov (United States)

Jones, M. B.; Slaughter, P.; Walker, L.; Jones, C. S.; Missier, P.; Ludäscher, B.; Cao, Y.; McPhillips, T.; Schildhauer, M.

2015-12-01

Scientific provenance describes the authenticity, origin, and processing history of research products and promotes scientific transparency by detailing the steps in computational workflows that produce derived products. These products include papers, findings, input data, software products to perform computations, and derived data and visualizations. The geosciences community values this type of information, and, at least theoretically, strives to base conclusions on computationally replicable findings. In practice, capturing detailed provenance is laborious and thus has been a low priority; beyond a lab notebook describing methods and results, few researchers capture and preserve detailed records of scientific provenance. We have built tools for capturing and publishing provenance that integrate into analytical environments that are in widespread use by geoscientists (R and Matlab). These tools lower the barrier to provenance generation by automating capture of critical information as researchers prepare data for analysis, develop, test, and execute models, and create visualizations. The 'recordr' library in R and the `matlab-dataone` library in Matlab provide shared functions to capture provenance with minimal changes to normal working procedures. Researchers can capture both scripted and interactive sessions, tag and manage these executions as they iterate over analyses, and then prune and publish provenance metadata and derived products to the DataONE federation of archival repositories. Provenance traces conform to the ProvONE model extension of W3C PROV, enabling interoperability across tools and languages. The capture system supports fine-grained versioning of science products and provenance traces. By assigning global identifiers such as DOIs, reseachers can cite the computational processes used to reach findings. And, finally, DataONE has built a web portal to search, browse, and clearly display provenance relationships between input data, the software

Targeting Self-Binding Peptides as a Novel Strategy To Regulate Protein Activity and Function: A Case Study on the Proto-oncogene Tyrosine Protein Kinase c-Src.

Science.gov (United States)

Bai, Zhengya; Hou, Shasha; Zhang, Shilei; Li, Zhongyan; Zhou, Peng

2017-04-24

Previously, we have reported a new biomolecular phenomenon spanning between protein folding and binding, termed as self-binding peptides (SBPs), where a short peptide segment in monomeric protein functions as a molecular switch by dynamically binding to/unbinding from its cognate domain in the monomer (Yang et al. J. Chem. Inf. 2015, 55, 329-342). Here, we attempt to raise the SBP as a new class of druggable targets to regulate the biological activity and function of proteins. A case study was performed on the proto-oncogene nonreceptor tyrosine kinase, c-Src, which contains two SBPs that bind separately to SH3 and SH2 domains of the kinase. State-of-the-art molecular dynamics (MD) simulations and post binding energetics analysis revealed that disrupting the kinase-intramolecular interactions of SH3 and SH2 domains with their cognate SBP ligands can result in totally different effects on the structural dynamics of c-Src kinase architecture; targeting the SH2 domain unlocks the autoinhibitory form of the kinase-this is very similar to the pTyr527 dephosphorylation that functionally activates the kinase, whereas targeting the SH3 domain can only release the domain from the tightly packed kinase but has a moderate effect on the kinase activity. Subsequently, based on the cognate SBP sequence we computationally designed a number of SH2-binding phosphopeptides using a motif grafting strategy. Fluorescence polarization (FP) assay observed that most of the designed phosphopeptides have higher binding affinity to SH2 domain as compared to the native SBP segment (K d = 53 nM). Kinase assay identified a typical dose-response relationship of phosphopeptides against kinase activation, substantiating that disruption of SH2-SBP interaction can mimic c-Src dephosphorylation and activate the kinase. Two rationally designed phosphopeptides, namely EPQpYEEIEN and EPQpYEELEN, were determined as strong binders of SH2 domain (K d = 8.3 and 15 nM, respectively) and potent activators of

Wood structural differences between northern and southern beech provenances growing at a moderate site.

Science.gov (United States)

Eilmann, B; Sterck, F; Wegner, L; de Vries, S M G; von Arx, G; Mohren, G M J; den Ouden, J; Sass-Klaassen, U

2014-08-01

Planting provenances originating from southern to northern locations has been discussed as a strategy to speed up species migration and mitigate negative effects of climate change on forest stability and productivity. Especially for drought-susceptible species such as European beech (Fagus sylvatica L.), the introduction of drought-tolerant provenances from the south could be an option. Yet, beech has been found to respond plastically to environmental conditions, suggesting that the climate on the plantation site might be more important for tree growth than the genetic predisposition of potentially drought-adapted provenances. In this study, we compared the radial growth, wood-anatomical traits and leaf phenology of four beech provenances originating from southern (Bulgaria, France) and northern locations (Sweden, the Netherlands) and planted in a provenance trial in the Netherlands. The distribution of precipitation largely differs between the sites of origin. The northern provenances experience a maximum and the southern provenances experience a minimum of rainfall in summer. We compared tree productivity and the anatomy of the water-conducting system for the period from 2000 to 2010, including the drought year 2003. In addition, tree mortality and the timing of leaf unfolding in spring were analysed for the years 2001, 2007 and 2012. Comparison of these traits in the four beech provenances indicates the influence of genetic predisposition and local environmental factors on the performance of these provenances under moderate site conditions. Variation in radial growth was controlled by environment, although the growth level slightly differed due to genetic background. The Bulgarian provenance had an efficient water-conducting system which was moreover unaffected by the drought in 2003, pointing to a high ability of this provenance to cope well with dry conditions. In addition, the Bulgarian provenance showed up as most productive in terms of height and radial

The constitutive activation of the CEF-4/9E3 chemokine gene depends on C/EBPbeta in v-src transformed chicken embryo fibroblasts

DEFF Research Database (Denmark)

Gagliardi, M; Maynard, S; Bojovic, B

2001-01-01

The CEF-4/9E3 chemokine gene is expressed constitutively in chicken embryo fibroblasts (CEF) transformed by the Rous sarcoma virus (RSV). This aberrant induction is controlled at the transcriptional and post-transcriptional levels. Transcriptional activation depends on multiple elements of the CEF....../EBPbeta binds to a second element located in proximity of the TRE. A mutation of this distal CAAT box impaired the activation of the CEF-4 promoter by pp60(v-src) indicating that this element is also part of the SRU. Using the RCASBP retroviral vector, we expressed a dominant negative mutant of C....../EBPbeta (designated Delta184-C/EBPbeta) in RSV-transformed CEF. Delta184-C/EBPbeta decreased the accumulation of the CEF-4 mRNA and activation of the CEF-4 promoter by pp60(v-src). The induction of the Cox-2 gene (CEF-147) was also reduced by Delta184-C/EBPbeta. The effect of the dominant negative mutant was observed...

Insect Residue Contamination on Wing Leading Edge Surfaces: A Materials Investigation for Mitigation

Science.gov (United States)

Lorenzi, Tyler M.; Wohl, Christopher J.; Penner, Ronald K.; Smith, Joseph G.; Siochi, Emilie J.

2011-01-01

Flight tests have shown that residue from insect strikes on aircraft wing leading edge surfaces may induce localized transition of laminar to turbulent flow. The highest density of insect populations have been observed between ground level and 153 m during light winds (2.6 -- 5.1 m/s), high humidity, and temperatures from 21 -- 29 C. At a critical residue height, dependent on the airfoil and Reynolds number, boundary layer transition from laminar to turbulent results in increased drag and fuel consumption. Although this represents a minimal increase in fuel burn for conventional transport aircraft, future aircraft designs will rely on maintaining laminar flow across a larger portion of wing surfaces to reduce fuel burn during cruise. Thus, insect residue adhesion mitigation is most critical during takeoff and initial climb to maintain laminar flow in fuel-efficient aircraft configurations. Several exterior treatments investigated to mitigate insect residue buildup (e.g., paper, scrapers, surfactants, flexible surfaces) have shown potential; however, implementation has proven to be impractical. Current research is focused on evaluation of wing leading edge surface coatings that may reduce insect residue adhesion. Initial work under NASA's Environmentally Responsible Aviation Program focused on evaluation of several commercially available products (commercial off-the-shelf, COTS), polymers, and substituted alkoxy silanes that were applied to aluminum (Al) substrates. Surface energies of these coatings were determined from contact angle data and were correlated to residual insect excrescence on coated aluminum substrates using a custom-built "bug gun." Quantification of insect excrescence surface coverage was evaluated by a series of digital photographic image processing techniques.

Ganoderiol A-enriched extract suppresses migration and adhesion of MDA-MB-231 cells by inhibiting FAK-SRC-paxillin cascade pathway.

Directory of Open Access Journals (Sweden)

Guo-Sheng Wu

Full Text Available Cell adhesion, migration and invasion are critical steps for carcinogenesis and cancer metastasis. Ganoderma lucidum, also called Lingzhi in China, is a traditional Chinese medicine, which exhibits anti-proliferation, anti-inflammation and anti-metastasis properties. Herein, GAEE, G. lucidum extract mainly contains ganoderiol A (GA, dihydrogenated GA and GA isomer, was shown to inhibit the abilities of adhesion and migration, while have a slight influence on that of invasion in highly metastatic breast cancer MDA-MB-231 cells at non-toxic doses. Further investigation revealed that GAEE decreased the active forms of focal adhesion kinase (FAK and disrupted the interaction between FAK and SRC, which lead to deactivating of paxillin. Moreover, GAEE treatment downregulated the expressions of RhoA, Rac1, and Cdc42, and decreased the interaction between neural Wiskott-Aldrich Syndrome protein (N-WASP and Cdc42, which impair cell migration and actin assembly. To our knowledge, this is the first report to show that G.lucidum triterpenoids could suppress cell migration and adhesion through FAK-SRC-paxillin signaling pathway. Our study also suggests that GAEE may be a potential agent for treatment of breast cancer.

Water use efficiency studies of Acacia senegal (L.) Willd provenances in Sudan

International Nuclear Information System (INIS)

Mustafa, A.F.; Elamin, K.H.; Salih, A.A.

1996-01-01

An experiment was conducted in 1989 to screen Acacia senegal L. Willd provenances collected from within the natural gum belt for high water use efficiency. Thirteen provenances were tested for water use efficiency and consequently 6 out of them were selected for further screening. The selection was based on their performance in the preliminary screening. Both the preliminary and the detailed study revealed that provenances 7, 3 and 11 combine high dry matter production with high water use efficiency. Water use efficiency and dry matter production appears to be negatively correlated with root length density and root/shoot ratios. Provenances 7 which exhibited the highest water use efficiency and dry matter yield had the lowest root/shoot ratio and also a low root length density. Based on these studies provenance 7 can be considered a suitable candidate for introduction into gum-belt of Sudan through for rehabilitation of this region. (author). 5 refs, 1 fig., 3 tabs

Water use efficiency studies of Acacia senegal (L.) Willd provenances in Sudan

Energy Technology Data Exchange (ETDEWEB)

Mustafa, A F; Elamin, K H [Forestry Research Section, Wad Medani (Sudan); Salih, A A [Soil Science Section, Wad Medani (Sudan)

1996-07-01

An experiment was conducted in 1989 to screen Acacia senegal L. Willd provenances collected from within the natural gum belt for high water use efficiency. Thirteen provenances were tested for water use efficiency and consequently 6 out of them were selected for further screening. The selection was based on their performance in the preliminary screening. Both the preliminary and the detailed study revealed that provenances 7, 3 and 11 combine high dry matter production with high water use efficiency. Water use efficiency and dry matter production appears to be negatively correlated with root length density and root/shoot ratios. Provenances 7 which exhibited the highest water use efficiency and dry matter yield had the lowest root/shoot ratio and also a low root length density. Based on these studies provenance 7 can be considered a suitable candidate for introduction into gum-belt of Sudan through for rehabilitation of this region. (author). 5 refs, 1 fig., 3 tabs.

Nonalbumin proteinuria predominates in biopsy-proven tenofovir nephrotoxicity.

Science.gov (United States)

Sise, Meghan E; Hirsch, Jamie S; Canetta, Pietro A; Herlitz, Leal; Mohan, Sumit

2015-05-15

Tenofovir disoproxil fumarate (TDF) nephrotoxicity is characterized by proximal renal tubular injury and dysmorphic mitochondria resulting in proteinuria, orthoglycemic glycosuria, and other markers of proximal tubular dysfunction. The objective of this study was to determine the pattern of proteinuria in patients with biopsy-proven TDF nephrotoxicity. Retrospective chart review. Patients with biopsy-proven TDF nephrotoxicity were identified and their medical charts and biopsy reports were reviewed. Comparison was made with HIV-infected patients not on TDF who underwent kidney biopsy. We identified 43 biopsy-proven cases of TDF nephrotoxicity; mean age 54.7 ± 0.4 years, 53% men, 42% whites. Thirty-seven cases reported proteinuria by dipstick of which only 60% had at least 2+ proteinuria. Twenty-seven patients had urine protein quantified by either 24-h collection or spot urine protein-to-creatinine ratio; median proteinuria was 1742 mg/day [interquartile range (IQR) 1200-2000 mg] and 1667 mg/g creatinine (IQR 851-1967 mg/g), respectively. Ten patients had concurrent urinary albumin measured, with a median 236 mg/g creatinine (IQR 137-343 mg/g). The mean urine albumin-to-urine protein ratio (uAPR) was 0.17 (IQR 0.14-0.19), confirming that TDF nephrotoxicity is primarily associated with nonalbumin proteinuria. Control cases had a uAPR of 0.65 (IQR 0.55-0.79) P < 0.001. Histopathology showed the predominance of proximal tubular injury with characteristic mitochondrial abnormalities. In the largest published cohort of patients with biopsy-proven TDF nephrotoxicity, we show that low uAPR is a reliable feature of this disease. Because of the predominance of nonalbumin proteinuria, dipstick urinalysis may be unreliable in TDF nephrotoxicity.

Identification of QTLs for resistance to sclerotinia stem rot and BnaC.IGMT5.a as a candidate gene of the major resistant QTL SRC6 in Brassica napus.

Directory of Open Access Journals (Sweden)

Jian Wu

Full Text Available Stem rot caused by Sclerotinia sclerotiorum in many important dicotyledonous crops, including oilseed rape (Brassica napus, is one of the most devastating fungal diseases and imposes huge yield loss each year worldwide. Currently, breeding for Sclerotinia resistance in B. napus, as in other crops, can only rely on germplasms with quantitative resistance genes. Thus, the identification of quantitative trait locus (QTL for S. sclerotiorum resistance/tolerance in this crop holds immediate promise for the genetic improvement of the disease resistance. In this study, ten QTLs for stem resistance (SR at the mature plant stage and three QTLs for leaf resistance (LR at the seedling stage in multiple environments were mapped on nine linkage groups (LGs of a whole genome map for B. napus constructed with SSR markers. Two major QTLs, LRA9 on LG A9 and SRC6 on LG C6, were repeatedly detected across all environments and explained 8.54-15.86% and 29.01%-32.61% of the phenotypic variations, respectively. Genotypes containing resistant SRC6 or LRA9 allele showed a significant reduction in disease lesion after pathogen infection. Comparative mapping with Arabidopsis and data mining from previous gene profiling experiments identified that the Arabidopsis homologous gene of IGMT5 (At1g76790 was related to the SRC6 locus. Four copies of the IGMT5 gene in B. napus were isolated through homologous cloning, among which, only BnaC.IGMT5.a showed a polymorphism between parental lines and can be associated with the SRC6. Furthermore, two parental lines exhibited a differential expression pattern of the BnaC.IGMT5.a gene in responding to pathogen inoculation. Thus, our data suggested that BnaC.IGMT5.a was very likely a candidate gene of this major resistance QTL.

Changing provenance of late Cenozoic sediments in the Jianghan Basin

Directory of Open Access Journals (Sweden)

Lei Shao

2015-07-01

Full Text Available The Yangtze River is one of the most important components of the East Asia river system. In this study, sediments in the Jianghan Basin, middle Yangtze River, were selected for trace element and rare earth element (REE measurements, in order to decipher information on the change of sediment provenance and evolution of the Yangtze River. According to the elemental variations, the late Cenozoic sediments of the Jianghan Basin could be divided into four parts. During 2.68–2.28 Ma and 1.25–0 Ma, provenance of the sediments was consistent, whereas sediments were derived from variable sources during 2.28–1.25 Ma. Comparison of the elemental compositions between the Pliocene and Quaternary sediments revealed a change in sediment source from a more felsic source area to a more basic source area around the Pliocene–Quaternary boundary. Input from the Emeishan LIP should account for this provenance change. Based on the provenance analysis of sediments in the Jianghan Basin, we infer that the Yangtze River developed into a large river with its drainage basin extended to the Emeishan LIP no later than the Pliocene–Quaternary boundary.

Mechanical Behavior Analysis of Y-Type S-SRC Column in a Large-Space Vertical Hybrid Structure Using Local Fine Numerical Simulation Method

Directory of Open Access Journals (Sweden)

Jianguang Yue

2018-01-01

Full Text Available In a large spatial structure, normally the important members are of special type and are the safety key for the global structure. In order to study the mechanical behavior details of the local member, it is difficult for the common test method to realize the complex spatial loading state of the local member. Therefore, a local-fine finite element model was proposed and a large-space vertical hybrid structure was numerically simulated. The seismic responses of the global structure and the Y-type S-SRC column were analyzed under El Centro seismic motions with the peak acceleration of 35 gal and 220 gal. The numerical model was verified with the results of the seismic shaking table test of the structure model. The failure mechanism and stiffness damage evolution of the Y-type S-SRC column were analyzed. The calculated results agreed well with the test results. It indicates that the local-fine FEM could reflect the mechanical details of the local members in a large spatial structure.

MBNL142 and MBNL143 gene isoforms, overexpressed in DM1-patient muscle, encode for nuclear proteins interacting with Src family kinases.

Science.gov (United States)

Botta, A; Malena, A; Tibaldi, E; Rocchi, L; Loro, E; Pena, E; Cenci, L; Ambrosi, E; Bellocchi, M C; Pagano, M A; Novelli, G; Rossi, G; Monaco, H L; Gianazza, E; Pantic, B; Romeo, V; Marin, O; Brunati, A M; Vergani, L

2013-08-15

Myotonic dystrophy type-1 (DM1) is the most prevalent form of muscular dystrophy in adults. This disorder is an RNA-dominant disease, caused by expansion of a CTG repeat in the DMPK gene that leads to a misregulation in the alternative splicing of pre-mRNAs. The longer muscleblind-like-1 (MBNL1) transcripts containing exon 5 and the respective protein isoforms (MBNL142-43) were found to be overexpressed in DM1 muscle and localized exclusively in the nuclei. In vitro assays showed that MBNL142-43 bind the Src-homology 3 domain of Src family kinases (SFKs) via their proline-rich motifs, enhancing the SFK activity. Notably, this association was also confirmed in DM1 muscle and myotubes. The recovery, mediated by an siRNA target to Ex5-MBNL142-43, succeeded in reducing the nuclear localization of both Lyn and MBNL142-43 proteins and in decreasing the level of tyrosine phosphorylated proteins. Our results suggest an additional molecular mechanism in the DM1 pathogenesis, based on an altered phosphotyrosine signalling pathway.

An Inference-Based Framework to Manage Data Provenance in Geoscience Applications

NARCIS (Netherlands)

Huq, M.R.; Apers, Peter M.G.; Wombacher, Andreas

2013-01-01

Data provenance allows scientists to validate their model as well as to investigate the origin of an unexpected value. Furthermore, it can be used as a replication recipe for output data products. However, capturing provenance requires enormous effort by scientists in terms of time and training.

Cell surface expression of channel catfish leukocyte immune-type receptors (IpLITRs) and recruitment of both Src homology 2 domain-containing protein tyrosine phosphatase (SHP)-1 and SHP-2.

Science.gov (United States)

Montgomery, Benjamin C S; Mewes, Jacqueline; Davidson, Chelsea; Burshtyn, Deborah N; Stafford, James L

2009-04-01

Channel catfish leukocyte immune-type receptors (IpLITRs) are immunoglobulin superfamily (IgSF) members believed to play a role in the control and coordination of cellular immune responses in teleost. Putative stimulatory and inhibitory IpLITRs are co-expressed by different types of catfish immune cells (e.g. NK cells, T cells, B cells, and macrophages) but their signaling potential has not been determined. Following cationic polymer-mediated transfections into human cell lines we examined the surface expression, tyrosine phosphorylation, and phosphatase recruitment potential of two types of putative inhibitory IpLITRs using 'chimeric' expression constructs and an epitope-tagged 'native' IpLITR. We also cloned and expressed the teleost Src homology 2 domain-containing protein tyrosine phosphatases (SHP)-1 and SHP-2 and examined their expression in adult tissues and developing zebrafish embryos. Co-immunoprecipitation experiments support the inhibitory signaling potential of distinct IpLITR-types that bound both SHP-1 and SHP-2 following the phosphorylation of tyrosine residues within their cytoplasmic tail (CYT) regions. Phosphatase recruitment by IpLITRs represents an important first step in understanding their influence on immune cell effector functions and suggests that certain inhibitory signaling pathways are conserved among vertebrates.

Provenance for Runtime Workflow Steering and Validation in Computational Seismology

Science.gov (United States)

Spinuso, A.; Krischer, L.; Krause, A.; Filgueira, R.; Magnoni, F.; Muraleedharan, V.; David, M.

2014-12-01

Provenance systems may be offered by modern workflow engines to collect metadata about the data transformations at runtime. If combined with effective visualisation and monitoring interfaces, these provenance recordings can speed up the validation process of an experiment, suggesting interactive or automated interventions with immediate effects on the lifecycle of a workflow run. For instance, in the field of computational seismology, if we consider research applications performing long lasting cross correlation analysis and high resolution simulations, the immediate notification of logical errors and the rapid access to intermediate results, can produce reactions which foster a more efficient progress of the research. These applications are often executed in secured and sophisticated HPC and HTC infrastructures, highlighting the need for a comprehensive framework that facilitates the extraction of fine grained provenance and the development of provenance aware components, leveraging the scalability characteristics of the adopted workflow engines, whose enactment can be mapped to different technologies (MPI, Storm clusters, etc). This work looks at the adoption of W3C-PROV concepts and data model within a user driven processing and validation framework for seismic data, supporting also computational and data management steering. Validation needs to balance automation with user intervention, considering the scientist as part of the archiving process. Therefore, the provenance data is enriched with community-specific metadata vocabularies and control messages, making an experiment reproducible and its description consistent with the community understandings. Moreover, it can contain user defined terms and annotations. The current implementation of the system is supported by the EU-Funded VERCE (http://verce.eu). It provides, as well as the provenance generation mechanisms, a prototypal browser-based user interface and a web API built on top of a NoSQL storage

Scientific Reproducibility in Biomedical Research: Provenance Metadata Ontology for Semantic Annotation of Study Description.

Science.gov (United States)

Sahoo, Satya S; Valdez, Joshua; Rueschman, Michael

2016-01-01

Scientific reproducibility is key to scientific progress as it allows the research community to build on validated results, protect patients from potentially harmful trial drugs derived from incorrect results, and reduce wastage of valuable resources. The National Institutes of Health (NIH) recently published a systematic guideline titled "Rigor and Reproducibility " for supporting reproducible research studies, which has also been accepted by several scientific journals. These journals will require published articles to conform to these new guidelines. Provenance metadata describes the history or origin of data and it has been long used in computer science to capture metadata information for ensuring data quality and supporting scientific reproducibility. In this paper, we describe the development of Provenance for Clinical and healthcare Research (ProvCaRe) framework together with a provenance ontology to support scientific reproducibility by formally modeling a core set of data elements representing details of research study. We extend the PROV Ontology (PROV-O), which has been recommended as the provenance representation model by World Wide Web Consortium (W3C), to represent both: (a) data provenance, and (b) process provenance. We use 124 study variables from 6 clinical research studies from the National Sleep Research Resource (NSRR) to evaluate the coverage of the provenance ontology. NSRR is the largest repository of NIH-funded sleep datasets with 50,000 studies from 36,000 participants. The provenance ontology reuses ontology concepts from existing biomedical ontologies, for example the Systematized Nomenclature of Medicine Clinical Terms (SNOMED CT), to model the provenance information of research studies. The ProvCaRe framework is being developed as part of the Big Data to Knowledge (BD2K) data provenance project.

Provenance variations of scots pine (Pinus sylvestris L.) in the Southern PART of Turkey

International Nuclear Information System (INIS)

Gulcu, S.; Bilir, N.

2015-01-01

Tree height, basal diameter, stem form, number, angle and diameter of branches were assessed in eight-year-old provenance test established by 30 seed sources of Scots pine (Pinus sylvestris L.) at Aydogmus and Kemer experimental sites of Southern part of Turkey. Growth of the provenances was also compared to two native species (Taurus cedar- Cedrus libani A. Rich and Black pine-Pinus nigra Arnold.) of the region. Variations within provenance and among provenances, and relations among the traits were estimated. There were large differences (p <= 0.05) within provenance and among provenances for the traits, while sites showed similar (0.05 <= p) performance for tree height and stem form. For instance, average of tree height was 181 cm and varied between 138.3 cm and 229.8 cm in provenances of Aydogmus site, it was 184 cm and ranged from 130 cm to 246.1 cm in that of Kemer site. Averages of tree height of a provenance were 144.4 cm in Aydogmus and 194.5 cm in Kemer. Individual tree height of the provenance varied between 69 cm and 267 cm, and ranged from 51 cm to 280 cm in sites. Averages of tree height were 143.2 cm in Black pine 145.6 cm in Taurus cedar which were natural species of the region. There were mostly positive and significant (p <= 0.05) correlations among the traits. Results of the study were discussed for new plantations and breeding of the species. (author)

PIXE study on the provenance of Chinese ancient porcelain

International Nuclear Information System (INIS)

Zhu, D.; Cheng, H.S.; Lin, J.W.; Yang, F.J.

2006-01-01

This paper reports the PIXE study on the provenance of Chinese ancient porcelain made in 7-10th century. The chemical compositions of Jun celadon samples made at Juntai, Linru and Hunyuan kilns, the white-glazed porcelain samples made at Ding, Huangye and Dangyangyu kilns, and the Ru celadon samples made at Qiangliang Temple were measured by external-beam PIXE, and the factor analysis was applied for identifying their provenances. Experimental results show that the porcelain products made at different kilns can be distinguished according to the compositional differences measured by PIXE

PIXE study on the provenance of Chinese ancient porcelain

Energy Technology Data Exchange (ETDEWEB)

Zhu, D. [Institute of Modern Physics, Fudan University, 220 Handan Road, Shanghai 200433 (China); Cheng, H.S. [Institute of Modern Physics, Fudan University, 220 Handan Road, Shanghai 200433 (China)]. E-mail: hscheng@fudan.edu.cn; Lin, J.W. [Institute of Modern Physics, Fudan University, 220 Handan Road, Shanghai 200433 (China); Yang, F.J. [Institute of Modern Physics, Fudan University, 220 Handan Road, Shanghai 200433 (China)

2006-08-15

This paper reports the PIXE study on the provenance of Chinese ancient porcelain made in 7-10th century. The chemical compositions of Jun celadon samples made at Juntai, Linru and Hunyuan kilns, the white-glazed porcelain samples made at Ding, Huangye and Dangyangyu kilns, and the Ru celadon samples made at Qiangliang Temple were measured by external-beam PIXE, and the factor analysis was applied for identifying their provenances. Experimental results show that the porcelain products made at different kilns can be distinguished according to the compositional differences measured by PIXE.

PIXE study on the provenance of Chinese ancient porcelain

Science.gov (United States)

Zhu, D.; Cheng, H. S.; Lin, J. W.; Yang, F. J.

2006-08-01

This paper reports the PIXE study on the provenance of Chinese ancient porcelain made in 7-10th century. The chemical compositions of Jun celadon samples made at Juntai, Linru and Hunyuan kilns, the white-glazed porcelain samples made at Ding, Huangye and Dangyangyu kilns, and the Ru celadon samples made at Qiangliang Temple were measured by external-beam PIXE, and the factor analysis was applied for identifying their provenances. Experimental results show that the porcelain products made at different kilns can be distinguished according to the compositional differences measured by PIXE.

Quadratic residues and non-residues selected topics

CERN Document Server

Wright, Steve

2016-01-01

This book offers an account of the classical theory of quadratic residues and non-residues with the goal of using that theory as a lens through which to view the development of some of the fundamental methods employed in modern elementary, algebraic, and analytic number theory. The first three chapters present some basic facts and the history of quadratic residues and non-residues and discuss various proofs of the Law of Quadratic Reciprosity in depth, with an emphasis on the six proofs that Gauss published. The remaining seven chapters explore some interesting applications of the Law of Quadratic Reciprocity, prove some results concerning the distribution and arithmetic structure of quadratic residues and non-residues, provide a detailed proof of Dirichlet’s Class-Number Formula, and discuss the question of whether quadratic residues are randomly distributed. The text is a valuable resource for graduate and advanced undergraduate students as well as for mathematicians interested in number theory.

Using Blockchain and smart contracts for secure data provenance management

OpenAIRE

Ramachandran, Aravind; Kantarcioglu, Dr. Murat

2017-01-01

Blockchain technology has evolved from being an immutable ledger of transactions for cryptocurrencies to a programmable interactive the environment for building distributed reliable applications. Although, blockchain technology has been used to address various challenges, to our knowledge none of the previous work focused on using blockchain to develop a secure and immutable scientific data provenance management framework that automatically verifies the provenance records. In this work, we le...

Provenance Representation in the Global Change Information System (GCIS)

Science.gov (United States)

Tilmes, Curt

2012-01-01

Global climate change is a topic that has become very controversial despite strong support within the scientific community. It is common for agencies releasing information about climate change to be served with Freedom of Information Act (FOIA) requests for everything that led to that conclusion. Capturing and presenting the provenance, linking to the research papers, data sets, models, analyses, observation instruments and satellites, etc. supporting key findings has the potential to mitigate skepticism in this domain. The U.S. Global Change Research Program (USGCRP) is now coordinating the production of a National Climate Assessment (NCA) that presents our best understanding of global change. We are now developing a Global Change Information System (GCIS) that will present the content of that report and its provenance, including the scientific support for the findings of the assessment. We are using an approach that will present this information both through a human accessible web site as well as a machine readable interface for automated mining of the provenance graph. We plan to use the developing W3C PROV Data Model and Ontology for this system.

Elevated expression of the cellular src gene in tumors of differing etiologies in Xiphophorus

Energy Technology Data Exchange (ETDEWEB)

Schartl, M.; Schmidt, C.R.; Anders, A.; Barnekow, A.

1985-08-15

In the fish Xiphophorus the authors have detected elevated levels of pp60c-src kinase activity in a variety of tumors (n = 34) of neurogenic, epithelial, and mesenchymal origin either of hereditary etiology or induced by carcinogens. This elevation ranged from 2-fold up to 50-fold compared to the corresponding non-tumorous tissue and up to 6-fold compared to the highest activities found in any of the normal organs. The level of elevation parallels the degree of malignancy in melanoma and in tumors of mesenchymal origin. In fish bearing tumors of hereditary etiology kinase activity was also elevated in the non-tumorous brain, while in fish bearing induced tumors, kinase activity was elevated only in the cells of the neoplasia.

Towards an Ontology-Driven Blockchain Design for Supply Chain Provenance

OpenAIRE

Kim, Henry M.; Laskowski, Marek

2016-01-01

An interesting research problem in our age of Big Data is that of determining provenance. Granular evaluation of provenance of physical goods--e.g. tracking ingredients of a pharmaceutical or demonstrating authenticity of luxury goods--has often not been possible with today's items that are produced and transported in complex, inter-organizational, often internationally-spanning supply chains. Recent adoption of Internet of Things and Blockchain technologies give promise at better supply chai...

Residual stress measurements in thick structural weldments by means of neutron diffraction

International Nuclear Information System (INIS)

Ohms, C.; Youtsos, A.G.; Idsert, P. v.d.; Timke, T.

2000-01-01

Welding residual stresses in large structural components are a major concern with respect to their performance and lifetime. In large structures reasonable thermal stress relief treatment is usually impossible due to the component size. On the other hand, prediction of welding stresses by numerical modelling has not yet proven to be generally reliable, while the experimental determination of such stresses remains a demanding task. At the high flux reactor (HFR), Petten, a new residual stress diffractometer has been installed recently capable of handling of components up to 1000 kg - the large component neutron diffraction facility (LCNDF). It has facilitated residual stress measurements in two large welded components, of which results are presented here. The first component represents a bi-metallic weld in form of a pipe of 25 mm wall thickness. Three dimensional measurements of residual stress are discussed in detail. The second specimen is a 66 mm wall thickness austenitic steel nuclear piping weld. Results on relief of strain within the weld through post weld heat treatment (PWHT) are presented. Additionally results obtained earlier at former CRNL (CAN) on a section of a thick nuclear piping weld are presented in order to illustrate the variation in the reference lattice parameter trough the weld and the heat affected zone (HAZ). These results clearly show the necessity to determine the reference parameters for each location in all measurement directions by means of measurements in small coupons free of macro-stresses. (orig.)

Biomarker-driven trial in metastatic pancreas cancer: feasibility in a multicenter study of saracatinib, an oral Src inhibitor, in previously treated pancreatic cancer

International Nuclear Information System (INIS)

Arcaroli, John; Quackenbush, Kevin; Dasari, Arvind; Powell, Rebecca; McManus, Martine; Tan, Aik-Choon; Foster, Nathan R; Picus, Joel; Wright, John; Nallapareddy, Sujatha; Erlichman, Charles; Hidalgo, Manuel; Messersmith, Wells A

2012-01-01

Src tyrosine kinases are overexpressed in pancreatic cancers, and the oral Src inhibitor saracatinib has shown antitumor activity in preclinical models of pancreas cancer. We performed a CTEP-sponsored Phase II clinical trial of saracatinib in previously treated pancreas cancer patients, with a primary endpoint of 6-month survival. A Simon MinMax two-stage phase II design was used. Saracatinib (175 mg/day) was administered orally continuously in 28-day cycles. In the unselected portion of the study, 18 patients were evaluable. Only two (11%) patients survived for at least 6 months, and three 6-month survivors were required to move to second stage of study as originally designed. The study was amended as a biomarker-driven trial (leucine rich repeat containing protein 19 [LRRC19] > insulin-like growth factor-binding protein 2 [IGFBP2] “top scoring pairs” polymerase chain reaction [PCR] assay, and PIK3CA mutant) based on preclinical data in a human pancreas tumor explant model. In the biomarker study, archival tumor tissue or fresh tumor biopsies were tested. Biomarker-positive patients were eligible for the study. Only one patient was PIK3CA mutant in a 3′ untranslated region (UTR) portion of the gene. This patient was enrolled in the study and failed to meet the 6-month survival endpoint. As the frequency of biomarker-positive patients was very low (<3%), the study was closed. Although we were unable to conclude whether enriching for a subset of second/third line pancreatic cancer patients treated with a Src inhibitor based on a biomarker would improve 6-month survival, we demonstrate that testing pancreatic tumor samples for a biomarker-driven, multicenter study in metastatic pancreas cancer is feasible

NGF-Dependent neurite outgrowth in PC12 cells overexpressing the Src homology 2-domain protein shb requires activation of the Rap1 pathway

NARCIS (Netherlands)

Lu, L.; Annerén, C.; Reedquist, K. A.; Bos, J. L.; Welsh, M.

2000-01-01

The Src homology 2 (SH2) domain adaptor protein Shb has been shown to transmit NGF- and FGF-2-dependent differentiation signals in PC12 cells. To study if this involves signaling through the small GTPase Rap1, Rap1 activity was assessed in Shb-overexpressing PC12 cells. We demonstrate that NGF and

Amphiregulin enhances VEGF-A production in human chondrosarcoma cells and promotes angiogenesis by inhibiting miR-206 via FAK/c-Src/PKCδ pathway.

Science.gov (United States)

Wang, Chao-Qun; Huang, Yu-Wen; Wang, Shih-Wei; Huang, Yuan-Li; Tsai, Chun-Hao; Zhao, Yong-Ming; Huang, Bi-Fei; Xu, Guo-Hong; Fong, Yi-Chin; Tang, Chih-Hsin

2017-01-28

Chondrosarcoma is the second most common primary malignancy of bone after myeloma and osteosarcoma. Chondrosarcoma development may be linked to angiogenesis, which is principally elicited by vascular endothelial growth factor-A (VEGF-A). The expression of VEGF-A has been recognized as a prognostic marker in angiogenesis. Amphiregulin (AR), an epidermal growth factor receptor ligand, promotes tumor proliferation, metastasis and angiogenesis. However, the role of AR in VEGF-A expression and angiogenesis in human chondrosarcoma remains largely unknown. This current study shows that AR promoted VEGF-A production and induced angiogenesis of human endothelial progenitor cells. Moreover, AR-enhanced VEGF-A expression and angiogenesis involved the FAK, c-Src and PKCδ signaling pathways, while miR-206 expression was negatively mediated by AR via the FAK, c-Src and PKCδ pathways. Our results illustrate the clinical significance between AR, VEGF-A and miR-206, as well as tumor stage, in human chondrosarcoma. AR may represent a novel therapeutic target in the metastasis and angiogenesis of chondrosarcoma. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

A New Cryogenic Sample Manipulator For SRC's Scienta 2002 System

International Nuclear Information System (INIS)

Gundelach, Chad T.; Fisher, Mike V.; Hoechst, Hartmut

2004-01-01

We discuss the first bench tests of a sample manipulator which was recently designed at SRC for the Scienta 2002 User system. The manipulator concept utilizes the 10 deg. angular window of the Scienta in the horizontal plane (angle dispersion) by rotating the sample normal around the vertical axis while angular scans along the vertical axis (energy dispersion) are continuous within ±30 deg. relative to the electron lens by rotating the sample around the horizontal axis. With this concept it is possible to precisely map the entire two-dimensional k-space of a crystal by means of stitching together 10 deg. wide stripes centered +15 deg. to -50 deg. relative to the sample normal. Three degrees of translational freedom allow positioning the sample surface at the focal point of the analyzer. Two degrees of rotational freedom are available at this position for manipulating the sample. Samples are mounted to a standard holder and transferred to the manipulator via a load-lock system attached to a prep chamber. The manipulator is configured with a cryogenic cold head, an electrical heater, and a temperature sensor permitting continuous closed-loop operation for 20-380 K

High basal Wnt signaling is further induced by PI3K/mTor inhibition but sensitive to cSRC inhibition in mammary carcinoma cell lines with HER2/3 overexpression

International Nuclear Information System (INIS)

Timmermans-Sprang, Elpetra P. M.; Gracanin, Ana; Mol, Jan A.

2015-01-01

Elevated basal, ligand-independent, Wnt signaling in some canine breast cancer cells is not caused by classical mutations in APC, β-Catenin or GSK3β but, at least partially, by enhanced LEF1 expression. We examined the expression and function of EGFR/HER-regulated pathways on the ligand-independent Wnt signaling. Twelve canine mammary tumor cell lines with previously reported differential basal Wnt activity were used. The expression levels of genes related to EGF-signaling were analyzed by cluster analysis. Cell lines with a combined overexpression of EGF-related genes and enhanced basal Wnt activity were treated with PI3K/mTor or cSRC inhibitors or transfected with a construct expressing wild-type PTEN. Subsequently, effects were measured on Wnt activity, cell proliferation, gene expression and protein level. High basal Wnt/LEF1 activity was associated with overexpression of HER2/3, ID1, ID2, RAC1 and HSP90 together with low to absent cMET and PTEN mRNA expression, suggesting a connection between Wnt- and HER-signaling pathways. Inhibition of the HER-regulated PI3K/mTor pathway using the dual PI3K/mTor inhibitor BEZ235 or the mTor inhibitor Everolimus® resulted in reduced cell proliferation. In the cell line with high basal Wnt activity, however, an unexpected further increased Wnt activity was found that could be greatly reduced after inhibition of the HER-regulated cSRC activity. Inhibition of the PI3K/mTor pathway was associated with enhanced expression of β-Catenin, Axin2, MUC1, cMET, EGFR and HER2 and a somewhat increased β-Catenin protein content, whereas cSRC inhibition was associated with slightly enhanced HER3 and SLUG mRNA expression. A high protein expression of HER3 was found only in a cell line with high basal Wnt activity. High basal Wnt activity in some mammary cancer cell lines is associated with overexpression of HER-receptor related genes and HER3 protein, and the absence of PTEN. Inhibition of the PI3K/mTor pathway further stimulated

2D QSAR studies of the inhibitory activity of a series of substituted purine derivatives against c-Src tyrosine kinase

OpenAIRE

Mukesh C. Sharma

2016-01-01

A series of 34 substituted purine analogues derivatives were subjected to quantitative structure-activity relationship analyses as inhibitors of c-Src tyrosine kinase. Partial least squares regression was applied to derive QSAR models, which were further validated for statistical significance by internal and external validation. The best QSAR model developed had a good predictive correlation coefficient (r2) of 0.8319, a significant cross-validated correlation coefficient (q2) of 0.7550, and ...

Data Provenance Inference in Logic Programming: Reducing Effort of Instance-driven Debugging

NARCIS (Netherlands)

Huq, M.R.; Mileo, Alessandra; Wombacher, Andreas

Data provenance allows scientists in different domains validating their models and algorithms to find out anomalies and unexpected behaviors. In previous works, we described on-the-fly interpretation of (Python) scripts to build workflow provenance graph automatically and then infer fine-grained

Non-destructive Inspection of Top-Down Construction Joints of Column in SRC Structure using Ultrasonic Method

International Nuclear Information System (INIS)

Park, Seok Kyun; Baek, Un Chan; Lee, Han Bum; Kim, Myoung Mo

2000-01-01

The joint treatment of concrete is one of the technical problems in top down construction method. Joints created with the top down construction result in serious weakness from the aspects of both structural and water-barrier function. Ultrasonic method was used for the inspection of top down construction joints of a various column in SRC structure in this study. The advantages and limitations of this method for non-destructive inspection in top down construction joints are investigated. As a result, it has been verified that the semi-direct measurement method is more effective than the other methods for detecting the voids of construction joints using ultrasonic method

Evolution of Src Homology 2 (SH2) Domain to Recognize Sulfotyrosine.

Science.gov (United States)

Ju, Tong; Niu, Wei; Guo, Jiantao

2016-09-16

Protein tyrosine O-sulfation is considered as the most common type of post-translational tyrosine modification in nature and plays important roles in extracellular biomolecular interactions. To facilitate the mapping, biological study, and medicinal application of this type of post-translational modification, we seek to evolve a small protein scaffold that recognizes sulfotyrosine with high affinity. We focused our efforts on the engineering of the Src Homology 2 (SH2) domain, which represents the largest class of known phosphotyrosine-recognition domain in nature and has a highly evolvable binding pocket. By using phage display, we successfully engineered the SH2 domain to recognize sulfotyrosine with high affinity. The best mutant, SH2-60.1, displayed more than 1700 fold higher sulfotyrosine-binding affinity than that of the wild-type SH2 domain. We also demonstrated that the evolved SH2 domain mutants could be used to detect sulfoprotein levels on the cell surface. These evolved SH2 domain mutants can be potentially applied to the study of protein tyrosine O-sulfation with proper experimental designs.

Provenance data in social media

CERN Document Server

Barbier, Geoffrey; Gundecha, Pritam

2013-01-01

Social media shatters the barrier to communicate anytime anywhere for people of all walks of life. The publicly available, virtually free information in social media poses a new challenge to consumers who have to discern whether a piece of information published in social media is reliable. For example, it can be difficult to understand the motivations behind a statement passed from one user to another, without knowing the person who originated the message. Additionally, false information can be propagated through social media, resulting in embarrassment or irreversible damages. Provenance data

Investigating Steroid Receptor Coactivator 3 (SRC3) as a Potential Therapeutic Target for Treating Advanced Prostate Cancer

Science.gov (United States)

2013-04-01

different kinases including MAPK, IKK, GSK3a , GSK3b , and CK1d. SRC-3 is also a target of ABL tyrosine kinase which can be activated by estrogen and...differentiated as evidenced by higher levels of Fkbp5, an AR-responsive gene that inhibits Akt signaling. These tumors also had lower levels of some...34 castrationCresistant" prostate" cancer,"we" found" that" although" androgen" deprivation" shrunk" the" size" of" the"tumor,"the"reduced" level "of"testosterone

Na,K-ATPase regulates intercellular communication in the vascular wall via cSrc kinase dependent connexin43 phosphorylation

DEFF Research Database (Denmark)

Hangaard, Lise; Bouzinova, Elena; Stæhr, Christian Albeck

2017-01-01

Communication between vascular smooth muscle cells (VSMCs) is dependent on gap junctions and is regulated by the Na-K-ATPase. The Na-K-ATPase is therefore important for synchronized VSMC oscillatory activity, i.e., vasomotion. The signaling between the Na-K-ATPase and gap junctions is unknown. We...... coupling in rat mesenteric small arteries in vitro. Phosphorylation of cSrc kinase and connexin43 (Cx43) were semiquantified by Western blotting. Micromole concentration of ouabain reduced the amplitude of norepinephrine-induced vasomotion and desynchronized Ca2+ transients in VSMC in the arterial wall...

Persistent identifiers for web service requests relying on a provenance ontology design pattern

Science.gov (United States)

Car, Nicholas; Wang, Jingbo; Wyborn, Lesley; Si, Wei

2016-04-01

Delivering provenance information for datasets produced from static inputs is relatively straightforward: we represent the processing actions and data flow using provenance ontologies and link to stored copies of the inputs stored in repositories. If appropriate detail is given, the provenance information can then describe what actions have occurred (transparency) and enable reproducibility. When web service-generated data is used by a process to create a dataset instead of a static inputs, we need to use sophisticated provenance representations of the web service request as we can no longer just link to data stored in a repository. A graph-based provenance representation, such as the W3C's PROV standard, can be used to model the web service request as a single conceptual dataset and also as a small workflow with a number of components within the same provenance report. This dual representation does more than just allow simplified or detailed views of a dataset's production to be used where appropriate. It also allow persistent identifiers to be assigned to instances of a web service requests, thus enabling one form of dynamic data citation, and for those identifiers to resolve to whatever level of detail implementers think appropriate in order for that web service request to be reproduced. In this presentation we detail our reasoning in representing web service requests as small workflows. In outline, this stems from the idea that web service requests are perdurant things and in order to most easily persist knowledge of them for provenance, we should represent them as a nexus of relationships between endurant things, such as datasets and knowledge of particular system types, as these endurant things are far easier to persist. We also describe the ontology design pattern that we use to represent workflows in general and how we apply it to different types of web service requests. We give examples of specific web service requests instances that were made by systems

Obsidian provenance research in the Americas.

Science.gov (United States)

Glascock, Michael D

2002-08-01

The characterization of archaeological materials to support provenance research has grown rapidly over the past few decades. Volcanic obsidian has several unique properties that make it the ideal archaeological material for studying prehistoric trade and exchange. This Account describes our laboratory's development of a systematic methodology for the characterization of obsidian sources and artifacts from Mesoamerica and other regions of North and South America in support of archaeological research.

Identification of provenance rocks based on EPMA analyses of heavy minerals

Science.gov (United States)

Shimizu, M.; Sano, N.; Ueki, T.; Yonaga, Y.; Yasue, K. I.; Masakazu, N.

2017-12-01

Information on mountain building is significant in the field of geological disposal of high-level radioactive waste, because this affects long-term stability in groundwater flow system. Provenance analysis is one of effective approaches for understanding building process of mountains. Chemical compositions of heavy minerals, as well as their chronological data, can be an index for identification of provenance rocks. The accurate identification requires the measurement of as many grains as possible. In order to achieve an efficient provenance analysis, we developed a method for quick identification of heavy minerals using an Electron Probe Micro Analyzer (EPMA). In this method, heavy mineral grains extracted from a sample were aligned on a glass slide and mounted in a resin. Concentration of 28 elements was measured for 300-500 grains per sample using EPMA. To measure as many grains as possible, we prioritized swiftness of measurement over precision, configuring measurement time of about 3.5 minutes for each grain. Identification of heavy minerals was based on their chemical composition. We developed a Microsoft® Excel® spread sheet input criteria of mineral identification using a typical range of chemical compositions for each mineral. The grains of 110 wt.% total were rejected. The criteria of mineral identification were revised through the comparison between mineral identification by optical microscopy and chemical compositions of grains classified as "unknown minerals". Provenance rocks can be identified based on abundance ratio of identified minerals. If no significant difference of the abundance ratio was found among source rocks, chemical composition of specific minerals was used as another index. This method was applied to the sediments of some regions in Japan where provenance rocks had lithological variations but similar formation ages. Consequently, the provenance rocks were identified based on chemical compositions of heavy minerals resistant to

Duox, Flotillin-2, and Src42A are required to activate or delimit the spread of the transcriptional response to epidermal wounds in Drosophila.

Directory of Open Access Journals (Sweden)

Michelle T Juarez

2011-12-01

Full Text Available The epidermis is the largest organ of the body for most animals, and the first line of defense against invading pathogens. A breach in the epidermal cell layer triggers a variety of localized responses that in favorable circumstances result in the repair of the wound. Many cellular and genetic responses must be limited to epidermal cells that are close to wounds, but how this is regulated is still poorly understood. The order and hierarchy of epidermal wound signaling factors are also still obscure. The Drosophila embryonic epidermis provides an excellent system to study genes that regulate wound healing processes. We have developed a variety of fluorescent reporters that provide a visible readout of wound-dependent transcriptional activation near epidermal wound sites. A large screen for mutants that alter the activity of these wound reporters has identified seven new genes required to activate or delimit wound-induced transcriptional responses to a narrow zone of cells surrounding wound sites. Among the genes required to delimit the spread of wound responses are Drosophila Flotillin-2 and Src42A, both of which are transcriptionally activated around wound sites. Flotillin-2 and constitutively active Src42A are also sufficient, when overexpressed at high levels, to inhibit wound-induced transcription in epidermal cells. One gene required to activate epidermal wound reporters encodes Dual oxidase, an enzyme that produces hydrogen peroxide. We also find that four biochemical treatments (a serine protease, a Src kinase inhibitor, methyl-ß-cyclodextrin, and hydrogen peroxide are sufficient to globally activate epidermal wound response genes in Drosophila embryos. We explore the epistatic relationships among the factors that induce or delimit the spread of epidermal wound signals. Our results define new genetic functions that interact to instruct only a limited number of cells around puncture wounds to mount a transcriptional response, mediating

Validation of quantitative method for azoxystrobin residues in green beans and peas.

Science.gov (United States)

Abdelraheem, Ehab M H; Hassan, Sayed M; Arief, Mohamed M H; Mohammad, Somaia G

2015-09-01

This study presents a method validation for extraction and quantitative analysis of azoxystrobin residues in green beans and peas using HPLC-UV and the results confirmed by GC-MS. The employed method involved initial extraction with acetonitrile after the addition of salts (magnesium sulfate and sodium chloride), followed by a cleanup step by activated neutral carbon. Validation parameters; linearity, matrix effect, LOQ, specificity, trueness and repeatability precision were attained. The spiking levels for the trueness and the precision experiments were (0.1, 0.5, 3 mg/kg). For HPLC-UV analysis, mean recoveries ranged between 83.69% to 91.58% and 81.99% to 107.85% for green beans and peas, respectively. For GC-MS analysis, mean recoveries ranged from 76.29% to 94.56% and 80.77% to 100.91% for green beans and peas, respectively. According to these results, the method has been proven to be efficient for extraction and determination of azoxystrobin residues in green beans and peas. Copyright © 2015 Elsevier Ltd. All rights reserved.

Physiological response to drought stress in Camptotheca acuminata seedlings from two provenances

Directory of Open Access Journals (Sweden)

Yeqing eYing

2015-05-01

Full Text Available Drought stress is a key environmental factor limiting the growth and productivity of plants. The purpose of this study was to investigate the physiological responses of Camptotheca acuminata (C. acuminata to different drought stresses and compare the drought tolerance between the provenances Kunming (KM and Nanchang (NC, which are naturally distributed in different rainfall zones with annual rainfalls of 1000-1100 mm and 1600-1700 mm, respectively. We determined relative water content (RWC, chlorophyll content (Chl(a+b, net photosynthesis (Pn, gas exchange parameters, relative leakage conductivity (REC, malondialdehyde (MDA content and superoxide dismutase (SOD and peroxidase (POD activities of C. acuminata seedlings under both moderate (50% of maximum field capacity and severe drought stress (30% of maximum field capacity. As the degree of water stress increased, RWC, Chl(a+b content, Pn, stomatal conductance (Gs, transpiration rate (Tr and intercellular CO2 concentration (Ci values decreased, but water use efficiency (WUE, REC, MDA content and SOD and POD activities increased in provenances KM and NC. Under moderate and severe drought stress, provenance KM had higher RWC, Chl(a+b, Pn, WUE, SOD and POD and lower Gs, Tr, Ci and REC in leaves than provenance NC. The results indicated that provenance KM may maintain stronger drought tolerance via improvements in water-retention capacity, antioxidant enzyme activity and membrane integrity.

Performance of Australian provenances of Eucalyptus grandis and Eucalyptus saligna in Hawaii

Science.gov (United States)

Roger G. Skolmen

1986-01-01

Australian provenances of Eucalyptus grandis and E. saligna were compared at four locations on the island of Hawaii to seek seed sources better than those in current use which were introduced earlier from unrecorded locations in Australia. A broad range of latitude and elevation was represented among the provenances. At all four...

The Src inhibitor dasatinib accelerates the differentiation of human bone marrow-derived mesenchymal stromal cells into osteoblasts

International Nuclear Information System (INIS)

Id Boufker, Hichame; Lagneaux, Laurence; Najar, Mehdi; Piccart, Martine; Ghanem, Ghanem; Body, Jean-Jacques; Journé, Fabrice

2010-01-01

The proto-oncogene Src is an important non-receptor protein tyrosine kinase involved in signaling pathways that control cell adhesion, growth, migration and differentiation. It negatively regulates osteoblast activity, and, as such, its inhibition is a potential means to prevent bone loss. Dasatinib is a new dual Src/Bcr-Abl tyrosine kinase inhibitor initially developed for the treatment of chronic myeloid leukemia. It has also shown promising results in preclinical studies in various solid tumors. However, its effects on the differentiation of human osteoblasts have never been examined. We evaluated the effects of dasatinib on bone marrow-derived mesenchymal stromal cells (MSC) differentiation into osteoblasts, in the presence or absence of a mixture of dexamethasone, ascorbic acid and β-glycerophosphate (DAG) for up to 21 days. The differentiation kinetics was assessed by evaluating mineralization of the extracellular matrix, alkaline phosphatase (ALP) activity, and expression of osteoblastic markers (receptor activator of nuclear factor kappa B ligand [RANKL], bone sialoprotein [BSP], osteopontin [OPN]). Dasatinib significantly increased the activity of ALP and the level of calcium deposition in MSC cultured with DAG after, respectively, 7 and 14 days; it upregulated the expression of BSP and OPN genes independently of DAG; and it markedly downregulated the expression of RANKL gene and protein (decrease in RANKL/OPG ratio), the key factor that stimulates osteoclast differentiation and activity. Our results suggest a dual role for dasatinib in both (i) stimulating osteoblast differentiation leading to a direct increase in bone formation, and (ii) downregulating RANKL synthesis by osteoblasts leading to an indirect inhibition of osteoclastogenesis. Thus, dasatinib is a potentially interesting candidate drug for the treatment of osteolysis through its dual effect on bone metabolism

Distal loop flexibility of a regulatory domain modulates dynamics and activity of C-terminal SRC kinase (csk.

Directory of Open Access Journals (Sweden)

Sulyman Barkho

Full Text Available The Src family of tyrosine kinases (SFKs regulate numerous aspects of cell growth and differentiation and are under the principal control of the C-terminal Src Kinase (Csk. Csk and SFKs share a modular design with the kinase domain downstream of the N-terminal SH2 and SH3 domains that regulate catalytic function and membrane localization. While the function of interfacial segments in these multidomain kinases are well-investigated, little is known about how surface sites and long-range, allosteric coupling control protein dynamics and catalytic function. The SH2 domain of Csk is an essential component for the down-regulation of all SFKs. A unique feature of the SH2 domain of Csk is the tight turn in place of the canonical CD loop in a surface site far removed from kinase domain interactions. In this study, we used a combination of experimental and computational methods to probe the importance of this difference by constructing a Csk variant with a longer SH2 CD loop to mimic the flexibility found in homologous kinase SH2 domains. Our results indicate that while the fold and function of the isolated domain and the full-length kinase are not affected by loop elongation, native protein dynamics that are essential for efficient catalysis are perturbed. We also identify key motifs and routes through which the distal SH2 site might influence catalysis at the active site. This study underscores the sensitivity of intramolecular signaling and catalysis to native protein dynamics that arise from modest changes in allosteric regions while providing a potential strategy to alter intrinsic activity and signaling modulation.

Novel autophosphorylation sites of Src family kinases regulate kinase activity and SH2 domain-binding capacity.

Science.gov (United States)

Weir, Marion E; Mann, Jacqueline E; Corwin, Thomas; Fulton, Zachary W; Hao, Jennifer M; Maniscalco, Jeanine F; Kenney, Marie C; Roman Roque, Kristal M; Chapdelaine, Elizabeth F; Stelzl, Ulrich; Deming, Paula B; Ballif, Bryan A; Hinkle, Karen L

2016-04-01

Src family tyrosine kinases (SFKs) are critical players in normal and aberrant biological processes. While phosphorylation importantly regulates SFKs at two known tyrosines, large-scale phosphoproteomics have revealed four additional tyrosines commonly phosphorylated in SFKs. We found these novel tyrosines to be autophosphorylation sites. Mimicking phosphorylation at the C-terminal site to the activation loop decreased Fyn activity. Phosphomimetics and direct phosphorylation at the three SH2 domain sites increased Fyn activity while reducing phosphotyrosine-dependent interactions. While 68% of human SH2 domains exhibit conservation of at least one of these tyrosines, few have been found phosphorylated except when found in cis to a kinase domain. © 2016 Federation of European Biochemical Societies.

Leaching characteristics of calcium-based compounds in MSWI Residues: From the viewpoint of clogging risk.

Science.gov (United States)

Xia, Yi; Zhang, Hua; Phoungthong, Khamphe; Shi, Dong-Xiao; Shen, Wen-Hui; Shao, Li-Ming; He, Pin-Jing

2015-08-01

Leachate collection system (LCS) clogging caused by calcium precipitation would be disadvantageous to landfill stability and operation. Meanwhile, calcium-based compounds are the main constituents in both municipal solid waste incineration bottom ash (MSWIBA) and stabilized air pollution control residues (SAPCR), which would increase the risk of LCS clogging once these calcium-rich residues were disposed in landfills. The leaching behaviors of calcium from the four compounds and municipal solid waste incineration (MSWI) residues were studied, and the influencing factors on leaching were discussed. The results showed that pH was the crucial factor in the calcium leaching process. CaCO3 and CaSiO3 began leaching when the leachate pH decreased to less than 7 and 10, respectively, while Ca3(PO4)2 leached at pHleaching rate for the different calcium-based compounds is as follows: CaSiO3>Ca3(PO4)2>CaCO3. The calcium leaching from the MSWIBA and SAPCR separately started from pHleaching respectively, which was proven by the X-ray diffraction results. Based on the leaching characteristics of the different calcium compounds and the mineral phase of calcium in the incineration residues, simulated computation of their clogging potential was conducted, providing the theoretical basis for the risk assessment pertaining to LCS clogging in landfills. Copyright © 2015 Elsevier Ltd. All rights reserved.

Detection of residual disease after neoadjuvant chemoradiotherapy for oesophageal cancer (preSANO): a prospective multicentre, diagnostic cohort study.

Science.gov (United States)

Noordman, Bo Jan; Spaander, Manon C W; Valkema, Roelf; Wijnhoven, Bas P L; van Berge Henegouwen, Mark I; Shapiro, Joël; Biermann, Katharina; van der Gaast, Ate; van Hillegersberg, Richard; Hulshof, Maarten C C M; Krishnadath, Kausilia K; Lagarde, Sjoerd M; Nieuwenhuijzen, Grard A P; Oostenbrug, Liekele E; Siersema, Peter D; Schoon, Erik J; Sosef, Meindert N; Steyerberg, Ewout W; van Lanschot, J Jan B

2018-05-31

After neoadjuvant chemoradiotherapy for oesophageal cancer, roughly half of the patients with squamous cell carcinoma and a quarter of those with adenocarcinoma have a pathological complete response of the primary tumour before surgery. Thus, the necessity of standard oesophagectomy after neoadjuvant chemoradiotherapy should be reconsidered for patients who respond sufficiently to neoadjuvant treatment. In this study, we aimed to establish the accuracy of detection of residual disease after neoadjuvant chemoradiotherapy with different diagnostic approaches, and the optimal combination of diagnostic techniques for clinical response evaluations. The preSANO trial was a prospective, multicentre, diagnostic cohort study at six centres in the Netherlands. Eligible patients were aged 18 years or older, had histologically proven, resectable, squamous cell carcinoma or adenocarcinoma of the oesophagus or oesophagogastric junction, and were eligible for potential curative therapy with neoadjuvant chemoradiotherapy (five weekly cycles of carboplatin [area under the curve 2 mg/mL per min] plus paclitaxel [50 mg/m 2 of body-surface area] combined with 41·4 Gy radiotherapy in 23 fractions) followed by oesophagectomy. 4-6 weeks after completion of neoadjuvant chemoradiotherapy, patients had oesophagogastroduodenoscopy with biopsies and endoscopic ultrasonography with measurement of maximum tumour thickness. Patients with histologically proven locoregional residual disease or no-pass during endoscopy and without distant metastases underwent immediate surgical resection. In the remaining patients a second clinical response evaluation was done (PET-CT, oesophagogastroduodenoscopy with biopsies, endoscopic ultrasonography with measurement of maximum tumour thickness, and fine-needle aspiration of suspicious lymph nodes), followed by surgery 12-14 weeks after completion of neoadjuvant chemoradiotherapy. The primary endpoint was the correlation between clinical response during

Structural and Functional Impacts of ER Coactivator Sequential Recruitment.

Science.gov (United States)

Yi, Ping; Wang, Zhao; Feng, Qin; Chou, Chao-Kai; Pintilie, Grigore D; Shen, Hong; Foulds, Charles E; Fan, Guizhen; Serysheva, Irina; Ludtke, Steven J; Schmid, Michael F; Hung, Mien-Chie; Chiu, Wah; O'Malley, Bert W

2017-09-07

Nuclear receptors recruit multiple coactivators sequentially to activate transcription. This "ordered" recruitment allows different coactivator activities to engage the nuclear receptor complex at different steps of transcription. Estrogen receptor (ER) recruits steroid receptor coactivator-3 (SRC-3) primary coactivator and secondary coactivators, p300/CBP and CARM1. CARM1 recruitment lags behind the binding of SRC-3 and p300 to ER. Combining cryo-electron microscopy (cryo-EM) structure analysis and biochemical approaches, we demonstrate that there is a close crosstalk between early- and late-recruited coactivators. The sequential recruitment of CARM1 not only adds a protein arginine methyltransferase activity to the ER-coactivator complex, it also alters the structural organization of the pre-existing ERE/ERα/SRC-3/p300 complex. It induces a p300 conformational change and significantly increases p300 HAT activity on histone H3K18 residues, which, in turn, promotes CARM1 methylation activity on H3R17 residues to enhance transcriptional activity. This study reveals a structural role for a coactivator sequential recruitment and biochemical process in ER-mediated transcription. Copyright © 2017 Elsevier Inc. All rights reserved.

Towards Provenance and Traceability in CRISTAL for HEP

CERN Document Server

Shamdasani, Jetendr; McClatchey, Richard

2014-01-01

This paper discusses the CRISTAL object lifecycle management system and its use in provenance data management and the traceability of system events. This software was initially used to capture the construction and calibration of the CMS ECAL detector at CERN for later use by physicists in their data analysis. Some further uses of CRISTAL in different projects (CMS, neuGRID and N4U) are presented as examples of its flexible data model. From these examples, applications are drawn for the High Energy Physics domain and some initial ideas for its use in data preservation HEP are outlined in detail in this paper. Currently investigations are underway to gauge the feasibility of using the N4U Analysis Service or a derivative of it to address the requirements of data and analysis logging and provenance capture within the HEP long term data analysis environment.

Surface plasmon resonance thermodynamic and kinetic analysis as a strategic tool in drug design. Distinct ways for phosphopeptides to plug into Src- and Grb2 SH2 domains

NARCIS (Netherlands)

de Mol, Nico J; Dekker, Frank J; Broutin, Isabel; Fischer, Marcel J E; Liskamp, Rob M J; Dekker, Frank

2005-01-01

Thermodynamic and kinetic studies of biomolecular interactions give insight into specificity of molecular recognition processes and advance rational drug design. Binding of phosphotyrosine (pY)-containing peptides to Src- and Grb2-SH2 domains was investigated using a surface plasmon resonance

Solvent refined coal (SRC) process. Flashing of SRC-II slurry in the vacuum column on Process Development Unit P-99. Interim report, February-June 1980

Energy Technology Data Exchange (ETDEWEB)

Gray, J. A.; Mathias, S. T.

1980-10-01

This report presents the results of 73 tests on the vacuum flash system of Process Development Unit P-99 performed during processing of three different coals; the second batch, fourth shipment (low ash batch) of Powhatan No. 5 Mine (LR-27383), Powhatan No. 6 Mine (LR-27596) and Ireland Mine (LR-27987). The objective of this work was to obtain experimental data for use in confirming and improving the design of the vacuum distillation column for the 6000 ton/day SRC-II Demonstration Plant. The 900/sup 0/F distillate content of the bottoms and the percent of feed flashed overhead were correlated with flash zone operating conditions for each coal, and the observed differences in performance were attributed to differences in the feed compositions. Retrogressive reactions appeared to be occurring in the 900/sup 0/F+ pyridine soluble material leading to an increase in the quantity of pyridine insoluble organic matter. Stream physical properties determined include specific gravity, viscosity and melting point. Elemental, distillation and solvent analyses were used to calculate component material balances. The Technology and Materials Department has used these results in a separate study comparing experimental K-values and vapor/liquid split with CHAMP computer program design predictions.

Proven Alternatives for Aboveground Treatment of Arsenic in Groundwater

Science.gov (United States)

This issue paper, developed for EPA's Engineering Forum, identifies and summarizes experiences with proven aboveground treatment alternatives for arsenic in groundwater, and provides information on their relative effectiveness and cost.

Residual stresses

International Nuclear Information System (INIS)

Sahotra, I.M.

2006-01-01

The principal effect of unloading a material strained into the plastic range is to create a permanent set (plastic deformation), which if restricted somehow, gives rise to a system of self-balancing within the same member or reaction balanced by other members of the structure., known as residual stresses. These stresses stay there as locked-in stresses, in the body or a part of it in the absence of any external loading. Residual stresses are induced during hot-rolling and welding differential cooling, cold-forming and extruding: cold straightening and spot heating, fabrication and forced fitting of components constraining the structure to a particular geometry. The areas which cool more quickly develop residual compressive stresses, while the slower cooling areas develop residual tensile stresses, and a self-balancing or reaction balanced system of residual stresses is formed. The phenomenon of residual stresses is the most challenging in its application in surface modification techniques determining endurance mechanism against fracture and fatigue failures. This paper discusses the mechanism of residual stresses, that how the residual stresses are fanned and what their behavior is under the action of external forces. Such as in the case of a circular bar under limit torque, rectangular beam under limt moment, reclaiming of shafts welds and peening etc. (author)

Bushen Huoxue Attenuates Diabetes-Induced Cognitive Impairment by Improvement of Cerebral Microcirculation: Involvement of RhoA/ROCK/moesin and Src Signaling Pathways

Directory of Open Access Journals (Sweden)

Yuan Li

2018-05-01

Full Text Available Type 2 Diabetes mellitus (T2DM is closely correlated with cognitive impairment and neurodegenerative disease. Bushen Huoxue (BSHX is a compound Chinese medicine used clinically to treat diabetes-induced cognitive impairment. However, its underlying mechanisms remain unclear. In the present study, KKAy mice, a genetic model of type 2 diabetes with obesity and insulin resistant hyperglycemia, received a daily administration of BSHX for 12 weeks. Blood glucose was measured every 4 weeks. After 12 weeks, BSHX treatment significantly ameliorated the T2DM related insults, including the increased blood glucose, the impaired spatial memory, decreased cerebral blood flow (CBF, occurrence of albumin leakage, leukocyte adhesion and opening capillary rarefaction. Meanwhile, the downregulation of the tight junction proteins (TJ claudin-5, occludin, zonula occluden-1 (ZO-1 and JAM-1 between endothelial cells, amyloid-β (Aβ accumulation in hippocampus, increased AGEs and RAGE, and expression of RhoA/ROCK/moesin signaling pathway and phosphorylation of Src kinase in KKAy mice were significantly protected by BSHX treatment. These results indicate that the protective effect of BSHX on T2DM-induced cognitive impairment involves regulation of RhoA/ROCK1/moesin signaling pathway and phosphorylation of Src kinase.

Proven commercial reactor types: an introduction to their principal advantages and disadvantages

International Nuclear Information System (INIS)

Alesso, H.P.

1981-01-01

This study deals with the principal advantages and disadvantages of the five types of proven commercial reactors. A description of each class of commercial reactor (light water, gas-cooled, and heavy water) and their proven reactors is followed by a comparison of reactor types on the basis of technical merit, economics of operation, availability of technology, and associated political issues. (author)

Solvent refined coal (SRC) process. Annual technical progress report, January 1979-December 1979

Energy Technology Data Exchange (ETDEWEB)

1980-11-01

This report discusses the effects on SRC yields of seven process variables (reactor temperature, SRT, hydrogen partial pressure, recycle ash and coal concentrations, gas velocity and coal type) predicted by second-order regression models developed from a data base containing pilot plant data with both Kentucky and Powhatan coals. The only effect of coal type in the model is a shift in each yield by a constant factor. Although some differences were found between the models developed from the Kentucky data base (1) (which we call Kentucky models) and the pooled coal models, the general conclusions of the previous report are confirmed by the new models and the assumption of similar behavior of the two coals appears to be justified. In some respects the dependence of the yields (MAF coal basis) on variables such as pressure and temperature are clearer than in the previous models. The principal trends which emerge are discussed.

Towards Automatic Capturing of Manual Data Processing Provenance

NARCIS (Netherlands)

Wombacher, Andreas; Huq, M.R.

2011-01-01

Often data processing is not implemented by a work ow system or an integration application but is performed manually by humans along the lines of a more or less specified procedure. Collecting provenance information during manual data processing can not be automated. Further, manual collection of

Scab susceptibility of a provenance collection of pecan

Science.gov (United States)

Scab (caused by Fusicladium effusum) is the most economically destructive disease of pecan in the Southeast US. Epidemics are favored by rainfall and high humidity. A provenance collection of ~950 pecan trees from 19 locations representing the native range of the species is located in Byron, Georgia...

EFFECTS OF COLD STRATIFICATION AND GA3 ON GERMINATION OF ARBUTUS UNEDO SEEDS OF THREE PROVENANCES.

Science.gov (United States)

Pipinis, Elias; Stampoulidis, Athanasios; Milios, Elias; Kitikidou, Kyriaki; Radoglou, Kalliopi

2017-01-01

Arbutus unedo is a valuable Mediterranean shrub as an ornamental plant as well as fruit tree. Fresh fruits of A. unedo are a good source of antioxidants, of vitamins C, E and carotenoids and also are characterized by the high content of mineral elements. The effects of gibberellic acid (GA 3 ) and cold stratification (CS) on seed germination performance were investigated in A. unedo seeds collected from three provenances in the Northern part of Greece. Seeds of each provenance were soaked in solutions of GA 3 (500, 1000 or 2000 ppm) for 24 h and subsequently were subjected to CS at 3 - 5°C for 0, 1, 2, and 3 months. Non-stratified seeds of the three A. unedo provenances which were not treated with GA 3 solutions exhibited very low germination. However, seed germination was significantly improved after a one-month period of CS. Similarly, the non-stratified seeds of all three provenances became non-dormant after the treatment with 2000 ppm GA 3 and they germinated at high percentages. However, in untreated seeds with GA3, after a one-month CS period the seeds of the Pieria provenance exhibited higher germination percentage than that of Rodopi provenance seeds. Furthermore, in non-stratified seeds, the Pieria provenance seeds treated with GA3 germinated at higher percentages and more rapidly than those of the other two provenances. The results indicated that untreated seeds exhibited very low germination at 20/25°C. However, in all three provenances seed germinability was significantly improved by a one-month period of CS or treatment of seeds with 2000 ppm GA3. Furthermore, there was a considerable variability among seed provenances in response to the treatments which were applied.

Growth and quality of Grevillea robusta provenances in Ruhande ...

African Journals Online (AJOL)

user

used as sawn timber for furniture industry, for construction and packaging. ... seven Queensland (Australia) provenances and one land race on an oxisol at Ruhande .... Statistical analyses were carried out using Statistical Analysis Systems.

Streptomyces thermoviolaceus SRC3 strain as a novel source of the antibiotic adjuvant streptazolin: A statistical approach toward the optimized production.

Science.gov (United States)

Djinni, Ibtissem; Djoudi, Warda; Souagui, Samiha; Rabia, Farida; Rahmouni, Sihem; Mancini, Ines; Kecha, Mouloud

2018-04-14

Streptomyces thermoviolaceus SRC3, a newly isolated actinobacterial strain from Algerian river sediments, exhibited a broad activity against various bacterial and yeast human pathogens (Salmonella Typhi ATCC 14028, Vibrio cholerae ATCC 14035, MRSA ATCC 43300 and Candida albicans ATCC 10231). The strain SRC3 was selected from thirty nine actinobacterial isolates and identified as S. thermoviolaceus based on morphology, cultural properties, physiological analyses and 16S rRNA gene sequencing. Culture parameters for the antibiotic production were optimized by sequential statistical strategy including Plackett-Burman design (PBD) and Response Surface Methodology (RSM). In PBD experiments, KCl, K 2 HPO 4 , MgSO 4 ·7H 2 O, pH value and incubation time emerged as the most significant in affecting the output of antimicrobial activities. These factors were further optimized using Central Composite Design (CCD). The best achieved conditions were: KCl (0.01%), K 2 HPO 4 (0.1%), MgSO 4 ·7H 2 O (0.02%) and 9 days incubation for anti-S. Typhi compounds, KCl (0.051%), MgSO 4 ·7H 2 O (0.05%) and 5 days incubation for C. albicans inhibitors. The metabolite responsible for the bioactivities was purified, structurally characterized (by NMR, MS, UV and IR analyses) and identified as streptazolin, recently reported as a promising antibiotic adjuvant. Copyright © 2018 Elsevier B.V. All rights reserved.

Providing Global Change Information for Decision-Making: Capturing and Presenting Provenance

Science.gov (United States)

Ma, Xiaogang; Fox, Peter; Tilmes, Curt; Jacobs, Katherine; Waple, Anne

2014-01-01

Global change information demands access to data sources and well-documented provenance to provide evidence needed to build confidence in scientific conclusions and, in specific applications, to ensure the information's suitability for use in decision-making. A new generation of Web technology, the Semantic Web, provides tools for that purpose. The topic of global change covers changes in the global environment (including alterations in climate, land productivity, oceans or other water resources, atmospheric composition and or chemistry, and ecological systems) that may alter the capacity of the Earth to sustain life and support human systems. Data and findings associated with global change research are of great public, government, and academic concern and are used in policy and decision-making, which makes the provenance of global change information especially important. In addition, since different types of decisions benefit from different types of information, understanding how to capture and present the provenance of global change information is becoming more of an imperative in adaptive planning.

Visualisation methods for large provenance collections in data-intensive collaborative platforms

Science.gov (United States)

Spinuso, Alessandro; Fligueira, Rosa; Atkinson, Malcolm; Gemuend, Andre

2016-04-01

This work investigates improving the methods of visually representing provenance information in the context of modern data-driven scientific research. It explores scenarios where data-intensive workflows systems are serving communities of researchers within collaborative environments, supporting the sharing of data and methods, and offering a variety of computation facilities, including HPC, HTC and Cloud. It focuses on the exploration of big-data visualization techniques aiming at producing comprehensive and interactive views on top of large and heterogeneous provenance data. The same approach is applicable to control-flow and data-flow workflows or to combinations of the two. This flexibility is achieved using the W3C-PROV recommendation as a reference model, especially its workflow oriented profiles such as D-PROV (Messier et al. 2013). Our implementation is based on the provenance records produced by the dispel4py data-intensive processing library (Filgueira et al. 2015). dispel4py is an open-source Python framework for describing abstract stream-based workflows for distributed data-intensive applications, developed during the VERCE project. dispel4py enables scientists to develop their scientific methods and applications on their laptop and then run them at scale on a wide range of e-Infrastructures (Cloud, Cluster, etc.) without making changes. Users can therefore focus on designing their workflows at an abstract level, describing actions, input and output streams, and how they are connected. The dispel4py system then maps these descriptions to the enactment platforms, such as MPI, Storm, multiprocessing. It provides a mechanism which allows users to determine the provenance information to be collected and to analyze it at runtime. For this work we consider alternative visualisation methods for provenance data, from infinite lists and localised interactive graphs, to radial-views. The latter technique has been positively explored in many fields, from text

Residual stresses

International Nuclear Information System (INIS)

Macherauch, E.

1978-01-01

Residual stresses are stresses which exist in a material without the influence of external powers and moments. They come into existence when the volume of a material constantly changes its form as a consequence of mechanical, thermal, and/or chemical processes and is hindered by neighbouring volumes. Bodies with residual stress are in mechanical balance. These residual stresses can be manifested by means of all mechanical interventions disturbing this balance. Acoustical, optical, radiological, and magnetical methods involving material changes caused by residual stress can also serve for determining residual stress. Residual stresses have an ambivalent character. In technical practice, they are feared and liked at the same time. They cause trouble because they can be the cause for unexpected behaviour of construction elements. They are feared since they can cause failure, in the worst case with catastrophical consequences. They are appreciated, on the other hand, because, in many cases, they can contribute to improvements of the material behaviour under certain circumstances. But they are especially liked for their giving convenient and (this is most important) mostly uncontrollable explanations. For only in very few cases we have enough knowledge and possibilities for the objective evaluation of residual stresses. (orig.) [de

Endothelium-Dependent Relaxation Effect of Apocynum venetum Leaf Extract via Src/PI3K/Akt Signalling Pathway

Directory of Open Access Journals (Sweden)

Yeh Siang Lau

2015-06-01

Full Text Available Botanical herbs are consumed globally not only as an essential diet but also as medicines or as functional/recreational food supplements. The extract of the Apocynum venetum leaves (AVLE, also known as Luobuma, exerts its antihypertensive effect via dilating the blood vessels in an endothelium- and concentration-dependent manner with optimal effect seen at as low as 10 µg/mL. A commercial Luoboma “antihypertensive tea” is available commercially in the western province of China. The present study seeks to investigate the underlying cellular mechanisms of the nitric oxide (NO-releasing property of AVLE in rat aortas and human umbilical vein endothelial cells (HUVECs. Endothelium-dependent relaxation induced by AVLE was assessed in organ chambers in the presence or absence of polyethyleneglycol catalase (PP2, 20 µM; inhibitor of Src kinase, wortmannin (30 nM and LY294002 (20 µM; PI3 (phosphatidylinositol3-Kinase inhibitor, NG-nitro-l-arginine (L-NAME, 100 µM; endothelial NO synthase inhibitor (eNOS and ODQ (1 µM; soluble guanylyl cyclase inhibitor. Total nitrite and nitrate (NOx level and protein expression of p-Akt and p-eNOS were measured. AVLE-induced endothelium-dependent relaxation was reduced by PP2, wortmannin and LY294002 and abolished by L-NAME and ODQ. AVLE significantly increased total NOx level in rat aortas and in HUVECs compared to control. It also instigated phosphorylation of Akt and eNOS in cultured HUVECs in a concentration-dependent manner and this was markedly suppressed by PP2, wortmannin and LY294002. AVLE also inhibited superoxide generated from both NADPH oxidase and xanthine/xanthine oxidase system. Taken together, AVLE causes endothelium-dependent NO mediated relaxations of rat aortas through Src/PI3K/Akt dependent NO signalling pathway and possesses superoxide scavenging activity.

Evaluation of residue-residue contact predictions in CASP9

KAUST Repository

Monastyrskyy, Bohdan

2011-01-01

This work presents the results of the assessment of the intramolecular residue-residue contact predictions submitted to CASP9. The methodology for the assessment does not differ from that used in previous CASPs, with two basic evaluation measures being the precision in recognizing contacts and the difference between the distribution of distances in the subset of predicted contact pairs versus all pairs of residues in the structure. The emphasis is placed on the prediction of long-range contacts (i.e., contacts between residues separated by at least 24 residues along sequence) in target proteins that cannot be easily modeled by homology. Although there is considerable activity in the field, the current analysis reports no discernable progress since CASP8.

Autumn frost hardiness in Norway spruce plus tree progeny and trees of the local and transferred provenances in central Sweden.

Science.gov (United States)

Hannerz, Mats; Westin, Johan

2005-09-01

Reforestation with provenances from locations remote from the planting site (transferred provenances) or the progeny of trees of local provenances selected for superior form and vigor (plus trees) offer alternative means to increase yield over that obtained by the use of seed from unselected trees of the local provenance. Under Swedish conditions, Norway spruce (Picea abies (L.) Karst.) of certain transferred provenances generally has an advantage in productivity relative to the local provenance comparable to that of progeny of plus trees. The aim of this study was to explore the extent to which productivity gains achieved by provenance transfer or the use of plus tree progeny are associated with reductions in autumn frost hardiness, relative to that of trees of the local provenance. In a field trial with 19-year-old trees in central Sweden, bud hardiness was tested on four occasions during the autumn of 2002. Trees of the local provenance were compared with trees of a south Swedish provenance originating 3 degrees of latitude to the south, a Belarusian provenance and the progeny of plus trees of local origin. The Belarusian provenance was the least hardy and the local provenance the most hardy, with plus tree progeny and the south Swedish provenance being intermediate in hardiness. Both the Belarusian provenance and the plus tree progeny were significantly taller than trees of the other populations. Within provenances, tree height was negatively correlated with autumn frost hardiness. Among the plus tree progeny, however, no such correlation between tree height and autumn frost hardiness was found. It is concluded that although the gain in productivity achieved by provenance transfer from Belarus was comparable to that achieved by using the progeny of plus trees of the local provenance, the use of trees of the Belarus provenance involved an increased risk of autumn frost damage because of later hardening.

Determined analysis of safety, viability and residual service life on criteria of crack mechanics

International Nuclear Information System (INIS)

Matvienko, Yu.G.

1997-01-01

Unified methods used in analysis of reliability, vulnerability, and residual lifetime of equipment with crack damage are considered, an increase in the desired lifetime is proven in the framework of vulnerability concept that allows crack developing with regard to the given level of reliability. The problem of reliability, vulnerability, and the lifetime is shown to be an interrelated problem. Optimal combination of the strength value, plasticity and resistance to crack developing results from the criteria of reliability and vulnerability based, in turn, on the principles of the mechanics of cracks. Structural features of technical systems can hinder the crack developing and prevent drastic damages of the equipment thus increasing the lifetime

Opening Up Climate Research: A Linked Data Approach to Publishing Data Provenance

Directory of Open Access Journals (Sweden)

Arif Shaon

2012-03-01

Full Text Available Traditionally, the formal scientific output in most fields of natural science has been limited to peer-reviewed academic journal publications, with less attention paid to the chain of intermediate data results and their associated metadata, including provenance. In effect, this has constrained the representation and verification of the data provenance to the confines of the related publications. Detailed knowledge of a dataset’s provenance is essential to establish the pedigree of the data for its effective re-use, and to avoid redundant re-enactment of the experiment or computation involved. It is increasingly important for open-access data to determine their authenticity and quality, especially considering the growing volumes of datasets appearing in the public domain. To address these issues, we present an approach that combines the Digital Object Identifier (DOI – a widely adopted citation technique – with existing, widely adopted climate science data standards to formally publish detailed provenance of a climate research dataset as an associated scientific workflow. This is integrated with linked-data compliant data re-use standards (e.g. OAI-ORE to enable a seamless link between a publication and the complete trail of lineage of the corresponding dataset, including the dataset itself.

Neutron activation analysis: a powerful tool in provenance investigations

International Nuclear Information System (INIS)

Meloni, Sandro; Oddone, Massimo

2002-01-01

It is well known that neutron activation analysis (NAA), both instrumental and destructive, allows the simultaneous determination of a number of elements, mostly trace elements, with high levels of precision and accuracy. These peculiar properties of NAA are very useful when applied to provenance studies, i.e. to the identification of the origin of raw materials with which artifacts had been manufactured in ancient times. Data reduction by statistical procedures, especially multivariate analysis techniques, provides a statistical 'fingerprint' of investigated materials, both raw materials and archaeological artifacts, that, upon comparison, allows the identification of the provenance of prime matters used for artifact manufacturing. Thus information on quarries and flows exploitation in the antiquity, on technological raw materials processing, on trade routes and about the circulation of fakes, can be obtained. In the present paper two case studies are reported. The first one deals with the identification of the provenance of clay used to make ceramic materials, mostly bricks and tiles, recovered from the excavation of a Roman 'villa' in Lomello (Roman name Laumellum) and of Roman settlings in Casteggio (Roman name Clastidium). Both sites are located in the Province of Pavia in areas called Lomellina and Oltrepo respectively. The second one investigates the origin of the white marble used to build medieval arks, Carolingian age, located in the church of San Felice, now property of the University of Pavia. Experimental set-up, analytical results and data reduction procedures are presented and discussed. (author)

Defining the "proven technology" technical criterion in the reactor technology assessment for Malaysia's nuclear power program

Science.gov (United States)

Anuar, Nuraslinda; Kahar, Wan Shakirah Wan Abdul; Manan, Jamal Abdul Nasir Abd

2015-04-01

Developing countries that are considering the deployment of nuclear power plants (NPPs) in the near future need to perform reactor technology assessment (RTA) in order to select the most suitable reactor design. The International Atomic Energy Agency (IAEA) reported in the Common User Considerations (CUC) document that "proven technology" is one of the most important technical criteria for newcomer countries in performing the RTA. The qualitative description of five desired features for "proven technology" is relatively broad and only provides a general guideline to its characterization. This paper proposes a methodology to define the "proven technology" term according to a specific country's requirements using a three-stage evaluation process. The first evaluation stage screens the available technologies in the market against a predefined minimum Technology Readiness Level (TRL) derived as a condition based on national needs and policy objectives. The result is a list of technology options, which are then assessed in the second evaluation stage against quantitative definitions of CUC desired features for proven technology. The potential technology candidates produced from this evaluation is further narrowed down to obtain a list of proven technology candidates by assessing them against selected risk criteria and the established maximum allowable total score using a scoring matrix. The outcome of this methodology is the proven technology candidates selected using an accurate definition of "proven technology" that fulfills the policy objectives, national needs and risk, and country-specific CUC desired features of the country that performs this assessment. A simplified assessment for Malaysia is carried out to demonstrate and suggest the use of the proposed methodology. In this exercise, ABWR, AP1000, APR1400 and EPR designs assumed the top-ranks of proven technology candidates according to Malaysia's definition of "proven technology".

Adjuvant Maneuvers for Residual Curvature Correction During Penile Prosthesis Implantation in Men with Peyronie's Disease.

Science.gov (United States)

Berookhim, Boback M; Karpman, Edward; Carrion, Rafael

2015-11-01

The surgical treatment of comorbid erectile dysfunction and Peyronie's disease has long included the implantation of an inflatable penile prosthesis as well as a number of adjuvant maneuvers to address residual curvature after prosthesis placement. To review the various surgical options for addressing curvature after prosthesis placement, with specific attention paid to an original article by Wilson et al. reporting on modeling over a penile prosthesis for the management of Peyronie's disease. A literature review was performed analyzing articles reporting the management of penile curvature in patients undergoing implantation of an inflatable penile prosthesis. Reported improvement in Peyronie's deformity as well as the complication rate associated with the various surgical techniques described. Modeling is a well-established treatment modality among patients with Peyronie's disease undergoing penile prosthesis implantation. A variety of other adjuvant maneuvers to address residual curvature when modeling alone is insufficient has been presented in the literature. Over 20 years of experience with modeling over a penile prosthesis have proven the efficacy and safety of this treatment option, providing the surgeon a simple initial step for the management of residual curvature after penile implantation which allows for the use of additional adjuvant maneuvers in those with significant deformities. © 2015 International Society for Sexual Medicine.

Characterization of automotive shredder residues before and five years after landfill disposal

Directory of Open Access Journals (Sweden)

Lucian Ionel Cioca

2015-04-01

Full Text Available The paper illustrates the results of an extensive analytical characterization study of automotive shredder residues (ASR, also known as "fluff”. The analyses concerned material fractions and their content, with special reference to heavy metals (e.g. Cd, Cr, Hg, Pb, Cu and arsenic. Elution tests on the original materials were also conducted. Moreover, chemical concentrations of ASR samples after about five years' landfill residence was assessed, in order to verify possible changes resulting from both in-situ leaching and organic matter degradation phenomena. Results show that lead seems to be the most critical element in view of possible ASR acceptance in non-hazardous waste landfills because of its high concentration in raw waste and, especially, of its proven leachability characteristics.

Isotopic provenance analysis and terrane tectonics: a warning about sediment transport distances

International Nuclear Information System (INIS)

Bassett, K.N.

1999-01-01

Full text: In the last 10 years the field of provenance analysis has undergone a revolution with the development of single-crystal isotopic dating techniques, the most common being U/Pb zircon and 40Ar/39Ar techniques. These have allowed age determination of single crystals thus providing more detail about probable provenance of each individual grain rather than an averaged population of grains. The usefulness for resolving complex terrane accretion and translation histories was immediately obvious and there have been many studies in many different regions aimed at tracking terrane motions by provenance of individual grains upward through the stratigraphy of a basin. Recent research in the North American Cordilleran terranes and in the New Zealand Torlesse Superterrane show how widely used and powerful these provenance analysis techniques are. However, isotopic provenance analysis has often been presented as key information to resolve controversies around terrane translation histories with very little discussion of the context of sedimentary facies and sediment transport mechanisms. An example is the recent use of U/Pb detrital zircon ages as the supposedly controversy-ending evidence for the amount of lateral translation of the Insular Superterrane in British Columbia (Baja BC) (Mahoney et al., 1999). The zircon grains were separated from fine-grained turbidite deposits and could easily have been transported over very large distances by a variety of mechanisms; yet they were presented as definitively resolving the Baja BC controversy. Modern examples illustrate the problem of using the provenance of fine grained sediment to constrain terrane tectonics. Sediment in the tip of the Bengal submarine fan was transported ∼3000 km from source, first by fluvial processes then by sediment gravity flow in the submarine fan. The detrital isotopic ages of single grains are the same as the depositional ages indicating a very rapid unroofing and transport rate with minimal

The impact of oil price on additions to US proven reserves

International Nuclear Information System (INIS)

Farzin, Y.H.

2001-01-01

Departing from Hotelling's assumption of fixed and known reserves, this paper develops an economic model of additions to proven reserves that explicitly incorporates the effects of expected resource price, cumulative reserves development, and technological progress on reserve additions. The model treats additions to proven oil reserves as output of a production process in which drilling wells is a primary input to transform some of oil-in-place into the economic category of proven reserves. Application of the model to US data for the 1950-1995 period provides strong statistical support for the existence of all the three salient effects. We obtain an estimate of the price elasticity of reserve additions (absent from previous studies) which, although statistically highly significant, is rather small. Using this price elasticity estimate, it is shown that if in the face of steady economic growth, and hence, oil consumption, US oil import dependence is to be kept from rising in the future, ceteris paribus, a steady oil price increase in the range of 1.5-4.5% a year is essential

A further evaluation of herbicides for post-emergence use in short rotation coppice

Energy Technology Data Exchange (ETDEWEB)

Turnbull, D.J.

2002-07-01

This report describes a study of the safety and efficacy of a range of herbicides and mixtures of herbicides (with both contact and residual activity) for the post-emergence control of weeds in newly planted willow short rotation coppice (SRC). Severe competition from weeds that have not been controlled adequately by pre-emergence herbicide application is one of the commonest causes of SRC crop failure. In the study, the effects of 11 herbicide treatments currently recommendation for weed control with cereals, legumes or potatoes were compared with an untreated control. There was minimal crop death from any treatment, though most of the treatments caused varying degrees of phytotoxicity. Two commercial products, Reflex T and Impuls, gave the best overall crop safety and weed control results. The report provides growers of SRC and their advisors with some information on how to achieve improved weed control in SRC fields, and recommends that British Biogen (the trade industry body) should consider the compilation of a technical register of herbicide applications based on information supplied by growers and advisers, including field treatment details.

Solvent refined coal studies: effects and characterization of treated solvent refined coal effluent. Progress report, FY 1977

Energy Technology Data Exchange (ETDEWEB)

Becker, C.D.; Woodfield, W.G.; Strand, J.A.

1978-07-01

The Freshwater Sciences Section of PNL has initiated biologically oriented studies at the P and M solvent refined coal (SRC) pilot plant on the Fort Lewis Reservation in western Washington. Essentially, the study objectives are to identify residual components in the treated SRC process and assess potential for adverse impact on water quality and aquatic biota. Since inception of research in mid-1976, six static toxicity tests with treated SRC process effluent have been conducted. Toxic components, not yet specifically identified, sometimes occur in the effluent. It is believed these components involve organic hydrocarbons of the phenol and cresol groups. Analyses have been obtained on inorganic and organic constituents in partially-treated and treated process effluent. Concentrations of inorganics identified in the effluent did not differ greatly from their concentrations in Lake Sequalitchew or SRC plant tap water, but the low concentrations may be due primarily to dilution with freshwater before discharge. Organics identified in the effluent are similar to those found in samples contaminated with petroleum, and involve many complex hydrocarbons.

Modification by covalent reaction or oxidation of cysteine residues in the tandem-SH2 domains of ZAP-70 and Syk can block phosphopeptide binding.

Science.gov (United States)

Visperas, Patrick R; Winger, Jonathan A; Horton, Timothy M; Shah, Neel H; Aum, Diane J; Tao, Alyssa; Barros, Tiago; Yan, Qingrong; Wilson, Christopher G; Arkin, Michelle R; Weiss, Arthur; Kuriyan, John

2015-01-01

Zeta-chain associated protein of 70 kDa (ZAP-70) and spleen tyrosine kinase (Syk) are non-receptor tyrosine kinases that are essential for T-cell and B-cell antigen receptor signalling respectively. They are recruited, via their tandem-SH2 (Src-homology domain 2) domains, to doubly phosphorylated immunoreceptor tyrosine-based activation motifs (ITAMs) on invariant chains of immune antigen receptors. Because of their critical roles in immune signalling, ZAP-70 and Syk are targets for the development of drugs for autoimmune diseases. We show that three thiol-reactive small molecules can prevent the tandem-SH2 domains of ZAP-70 and Syk from binding to phosphorylated ITAMs. We identify a specific cysteine residue in the phosphotyrosine-binding pocket of each protein (Cys39 in ZAP-70, Cys206 in Syk) that is necessary for inhibition by two of these compounds. We also find that ITAM binding to ZAP-70 and Syk is sensitive to the presence of H2O2 and these two cysteine residues are also necessary for inhibition by H2O2. Our findings suggest a mechanism by which the reactive oxygen species generated during responses to antigen could attenuate signalling through these kinases and may also inform the development of ZAP-70 and Syk inhibitors that bind covalently to their SH2 domains.

Drought effects on root and needle terpenoid content of a coastal and an interior Douglas fir provenance.

Science.gov (United States)

Kleiber, Anita; Duan, Qiuxiao; Jansen, Kirstin; Verena Junker, Laura; Kammerer, Bernd; Rennenberg, Heinz; Ensminger, Ingo; Gessler, Arthur; Kreuzwieser, Jürgen

2017-12-01

Douglas fir (Pseudotsuga menziesii) is a conifer species that stores large amounts of terpenoids, mainly monoterpenoids in resin ducts of various tissues. The effects of drought on stored leaf terpenoid concentrations in trees are scarcely studied and published data are partially controversial, since reduced, unaffected or elevated terpenoid contents due to drought have been reported. Even less is known on the effect of drought on root terpenoids. In the present work, we investigated the effect of reduced water availability on the terpenoid content in roots and needles of Douglas fir seedlings. Two contrasting Douglas fir provenances were studied: an interior provenance (var. glauca) with assumed higher drought resistance, and a coastal provenance (var. menziesii) with assumed lower drought resistance. We tested the hypothesis that both provenances show specific patterns of stored terpenoids and that the patterns will change in response to drought in both, needles and roots. We further expected stronger changes in the less drought tolerant coastal provenance. For this purpose, we performed an experiment under controlled conditions, in which the trees were exposed to moderate and severe drought stress. According to our expectations, the study revealed clear provenance-specific terpenoid patterns in needles. However, such patterns were not detected in the roots. Drought slightly increased the needle terpenoid contents of the coastal but not of the interior provenance. We also observed increased terpenoid abundance mainly in roots of the moderately stressed coastal provenance. Overall, from the observed provenance-specific reactions with increased terpenoid levels in trees of the coastal origin in response to drought, we conclude on functions of terpenoids for abiotic stress tolerance that might be fulfilled by other, constitutively expressed mechanisms in drought-adapted interior provenances. © The Author 2017. Published by Oxford University Press. All rights

SRC family kinase inhibitor SU6656 enhances antiangiogenic effect of irradiation

International Nuclear Information System (INIS)

Cuneo, Kyle C.; Geng Ling; Tan Jiahuai; Brousal, Jeffrey; Shinohara, Eric T.; Osusky, Katherine; Fu, Allie; Shyr, Yu; Wu Huiyun; Hallahan, Dennis E.

2006-01-01

Purpose: Src family kinases (SFK) have been identified as molecular targets. SU6656 is a small-molecule indolinone that specifically inhibits this family of kinases. Methods and Materials: Human umbilical vein endothelial cells were used to study the effects of SFK inhibition. Western blot analysis was performed to determine the effect of SFK inhibition on the PI3K/Akt pathway and caspase cleavage. Apoptosis was studied by propidium iodide staining of nuclei. Angiogenesis was examined using capillary tubule formation in Matrigel. Tumor response was further studied in vivo using Lewis lung carcinoma cells implanted into the dorsal skin fold of mice in the window model and in the hind limb in the tumor volume model. Results: Clonogenic survival of endothelial cells was decreased after the combined therapy of SU6656 and radiation compared with radiotherapy alone. Furthermore, SFK inhibition by SU6656 attenuated radiation-induced Akt phosphorylation and increased radiation-induced apoptosis and vascular endothelium destruction. In vivo, SU6656 administered before irradiation significantly enhanced radiation-induced destruction of blood vessels within the tumor windows and enhanced tumor growth delay when administered during fractionated irradiation. Conclusions: This study demonstrates the potential use of SFK inhibition to enhance the effects of ionizing radiation during radiotherapy

Residual stress measurements in a ferritic steel/In625 superalloy dissimilar metal weldment using neutron diffraction and deep-hole drilling

International Nuclear Information System (INIS)

Skouras, A.; Paradowska, A.; Peel, M.J.; Flewitt, P.E.J.; Pavier, M.J.

2013-01-01

This paper reports the use of non-invasive and semi-invasive techniques to measure the residual stresses in a large dissimilar weldment. This took the form of a butt weld between two sections of a P92 steel pipe, joined using an In625 welding consumable. Residual stress measurements have been carried out on the 30 mm thick welded pipe using the deep-hole drilling technique to characterise the through wall section residual stress distribution for the weld metal, HAZ and parent material. In addition, neutron diffraction measurements have been carried out within the weld zone. Diffraction patterns presented a high intensity and sharp peaks for the base P92 steel material. However measurements in the weld superalloy material were proven problematic as very weak diffraction patterns were observed. A thorough examination of the weld material suggested that the likely cause of this phenomenon was texture in the weld material created during the solidification phase of the welding procedure. This paper discusses the challenges in the execution and interpretation of the neutron diffraction results and demonstrates that realistic measurements of residual stresses can be achieved, in complex dissimilar metal weldments. Highlights: ► One of the few papers to measure residual stresses on dissimilar metal welds. ► Paper managed to provide realistic measurements of residual stresses using the DHD and ND technique. ► Results of this study have demonstrated the effect of texture during the ND measurements.

Provenance metadata gathering and cataloguing of EFIT++ code execution

Energy Technology Data Exchange (ETDEWEB)

Lupelli, I., E-mail: ivan.lupelli@ccfe.ac.uk [CCFE, Culham Science Centre, Abingdon, Oxon OX14 3DB (United Kingdom); Muir, D.G.; Appel, L.; Akers, R.; Carr, M. [CCFE, Culham Science Centre, Abingdon, Oxon OX14 3DB (United Kingdom); Abreu, P. [Instituto de Plasmas e Fusão Nuclear, Instituto Superior Técnico, Universidade de Lisboa, 1049-001 Lisboa (Portugal)

2015-10-15

Highlights: • An approach for automatic gathering of provenance metadata has been presented. • A provenance metadata catalogue has been created. • The overhead in the code runtime is less than 10%. • The metadata/data size ratio is about ∼20%. • A visualization interface based on Gephi, has been presented. - Abstract: Journal publications, as the final product of research activity, are the result of an extensive complex modeling and data analysis effort. It is of paramount importance, therefore, to capture the origins and derivation of the published data in order to achieve high levels of scientific reproducibility, transparency, internal and external data reuse and dissemination. The consequence of the modern research paradigm is that high performance computing and data management systems, together with metadata cataloguing, have become crucial elements within the nuclear fusion scientific data lifecycle. This paper describes an approach to the task of automatically gathering and cataloguing provenance metadata, currently under development and testing at Culham Center for Fusion Energy. The approach is being applied to a machine-agnostic code that calculates the axisymmetric equilibrium force balance in tokamaks, EFIT++, as a proof of principle test. The proposed approach avoids any code instrumentation or modification. It is based on the observation and monitoring of input preparation, workflow and code execution, system calls, log file data collection and interaction with the version control system. Pre-processing, post-processing, and data export and storage are monitored during the code runtime. Input data signals are captured using a data distribution platform called IDAM. The final objective of the catalogue is to create a complete description of the modeling activity, including user comments, and the relationship between data output, the main experimental database and the execution environment. For an intershot or post-pulse analysis (∼1000

Provenance metadata gathering and cataloguing of EFIT++ code execution

International Nuclear Information System (INIS)

Lupelli, I.; Muir, D.G.; Appel, L.; Akers, R.; Carr, M.; Abreu, P.

2015-01-01

Highlights: • An approach for automatic gathering of provenance metadata has been presented. • A provenance metadata catalogue has been created. • The overhead in the code runtime is less than 10%. • The metadata/data size ratio is about ∼20%. • A visualization interface based on Gephi, has been presented. - Abstract: Journal publications, as the final product of research activity, are the result of an extensive complex modeling and data analysis effort. It is of paramount importance, therefore, to capture the origins and derivation of the published data in order to achieve high levels of scientific reproducibility, transparency, internal and external data reuse and dissemination. The consequence of the modern research paradigm is that high performance computing and data management systems, together with metadata cataloguing, have become crucial elements within the nuclear fusion scientific data lifecycle. This paper describes an approach to the task of automatically gathering and cataloguing provenance metadata, currently under development and testing at Culham Center for Fusion Energy. The approach is being applied to a machine-agnostic code that calculates the axisymmetric equilibrium force balance in tokamaks, EFIT++, as a proof of principle test. The proposed approach avoids any code instrumentation or modification. It is based on the observation and monitoring of input preparation, workflow and code execution, system calls, log file data collection and interaction with the version control system. Pre-processing, post-processing, and data export and storage are monitored during the code runtime. Input data signals are captured using a data distribution platform called IDAM. The final objective of the catalogue is to create a complete description of the modeling activity, including user comments, and the relationship between data output, the main experimental database and the execution environment. For an intershot or post-pulse analysis (∼1000

A history-tracing XML-based provenance framework for workflows

NARCIS (Netherlands)

Gerhards, M; Belloum, A.; Berretz, F.; Sander, V.; Skorupa, S.

2010-01-01

The importance of validating and reproducing the outcome of computational processes is fundamental to many application domains. Assuring the provenance of workflows will likely become even more important with respect to the incorporation of human tasks to standard workflows by emerging standards

Effect of process conditions on the gel viscosity and gel strength of semi-refined carrageenan (SRC produced from seaweed (Kappaphycus alvarezii

Directory of Open Access Journals (Sweden)

Awang Bono

2014-01-01

Full Text Available Kappaphycus alvarezii or commonly known Euchema cottonii is a good source of kappa-carrageenan and can be found cultivated in the coastal areas of Malaysia, Philippines and Indonesia. Carrageenans have many applications and are utilized in human food and pet-food industry. Carrageenans are also utilized in non-food industry such as pharmaceuticals, cosmetics, printing and textile formulations. Currently, the Southeast Asian region is producing semi refined carrageenan (SRC. There are various works in producing SRC; however, there are limited efforts to develop the optimization of cooking process parameters. Hence, the present study features on the cooking process (alkaline treatment where the parameters (concentration of potassium hydroxide solution, cooking time and cooking temperature and the ranges are identified experimentally. The effects of these parameters on carrageenan quality such as gel viscosity and gel strength were studied. The optimization of cooking process parameters and the experimental design was conducted based on the Central Composite Design (CCD of Response Surface Methodology (RSM. The experimental result showed that gel viscosity increases with the decrease of cooking time, cooking temperature and potassium hydroxide (KOH concentration (% w/w. In contrast, gel strength increases as cooking time, cooking temperature and KOH concentration (% w/w increases. From the optimization, the best conditions for alkaline treatment found were cooking temperature 80 °C, cooking time 30 min and KOH concentration 10 (% w/w which are similar to current practice in industry.

S-ProvFlow: provenance model and tools for scalable and adaptive analysis pipelines in geoscience.

Science.gov (United States)

Spinuso, A.; Mihajlovski, A.; Atkinson, M.; Filgueira, R.; Klampanos, I.; Sanchez, S.

2017-12-01

The reproducibility of scientific findings is essential to improve the quality and application of modern data-driven research. Delivering such reproducibility is challenging in the context of systems handling large data-streams with sophisticated computational methods. Similarly, the SKA (Square Kilometer Array) will collect an unprecedented volume of radio-wave signals that will have to be reduced and transformed into derived products, with impact on space-weather research. This highlights the importance of having cross-disciplines mechanisms at the producer's side that rely on usable lineage data to support validation and traceability of the new artifacts. To be informative, provenance has to describe each methods' abstractions and their implementation as mappings onto distributed platforms and their concurrent execution, capturing relevant internal dependencies at runtime. Producers and intelligent toolsets should be able to exploit the produced provenance, steering real-time monitoring activities and inferring adaptations of methods at runtime.We present a model of provenance (S-PROV) that extends W3C PROV and ProvONE, broadening coverage of provenance to aspects related to distribution, scale-up and steering of stateful streaming operators in analytic pipelines. This is supported by a technical framework for tuneable and actionable lineage, ensuring its relevance to the users' interests, fostering its rapid exploitation to facilitate research practices. By applying concepts such as provenance typing and profiling, users define rules to capture common provenance patterns and activate selective controls based on domain-metadata. The traces are recorded in a document-store with index optimisation and a web API serves advanced interactive tools (S-ProvFlow, https://github.com/KNMI/s-provenance). These allow different classes of consumers to rapidly explore the provenance data. The system, which contributes to the SKA-Link initiative, within technology and

EGFR/Src/Akt signaling modulates Sox2 expression and self-renewal of stem-like side-population cells in non-small cell lung cancer.

Science.gov (United States)

Singh, Sandeep; Trevino, Jose; Bora-Singhal, Namrata; Coppola, Domenico; Haura, Eric; Altiok, Soner; Chellappan, Srikumar P

2012-09-25

Cancer stem cells are thought to be responsible for the initiation and progression of cancers. In non-small cell lung cancers (NSCLCs), Hoechst 33342 dye effluxing side population (SP) cells are shown to have stem cell like properties. The oncogenic capacity of cancer stem-like cells is in part due to their ability to self-renew; however the mechanistic correlation between oncogenic pathways and self-renewal of cancer stem-like cells has remained elusive. Here we characterized the SP cells at the molecular level and evaluated its ability to generate tumors at the orthotopic site in the lung microenvironment. Further, we investigated if the self-renewal of SP cells is dependent on EGFR mediated signaling. SP cells were detected and isolated from multiple NSCLC cell lines (H1650, H1975, A549), as well as primary human tumor explants grown in nude mice. SP cells demonstrated stem-like properties including ability to self-renew and grow as spheres; they were able to generate primary and metastatic tumors upon orthotopic implantation into the lung of SCID mice. In vitro study revealed elevated expression of stem cell associated markers like Oct4, Sox2 and Nanog as well as demonstrated intrinsic epithelial to mesenchymal transition features in SP cells. Further, we show that abrogation of EGFR, Src and Akt signaling through pharmacological or genetic inhibitors suppresses the self-renewal growth and expansion of SP-cells and resulted in specific downregulation of Sox2 protein expression. siRNA mediated depletion of Sox2 significantly blocked the SP phenotype as well as its self-renewal capacity; whereas other transcription factors like Oct4 and Nanog played a relatively lesser role in regulating self-renewal. Interestingly, Sox2 was elevated in metastatic foci of human NSCLC samples. Our findings suggest that Sox2 is a novel target of EGFR-Src-Akt signaling in NSCLCs that modulates self-renewal and expansion of stem-like cells from NSCLC. Therefore, the outcome of the

EGFR/Src/Akt signaling modulates Sox2 expression and self-renewal of stem-like side-population cells in non-small cell lung cancer

Directory of Open Access Journals (Sweden)

Singh Sandeep

2012-09-01

Full Text Available Abstract Background Cancer stem cells are thought to be responsible for the initiation and progression of cancers. In non-small cell lung cancers (NSCLCs, Hoechst 33342 dye effluxing side population (SP cells are shown to have stem cell like properties. The oncogenic capacity of cancer stem-like cells is in part due to their ability to self-renew; however the mechanistic correlation between oncogenic pathways and self-renewal of cancer stem-like cells has remained elusive. Here we characterized the SP cells at the molecular level and evaluated its ability to generate tumors at the orthotopic site in the lung microenvironment. Further, we investigated if the self-renewal of SP cells is dependent on EGFR mediated signaling. Results SP cells were detected and isolated from multiple NSCLC cell lines (H1650, H1975, A549, as well as primary human tumor explants grown in nude mice. SP cells demonstrated stem-like properties including ability to self-renew and grow as spheres; they were able to generate primary and metastatic tumors upon orthotopic implantation into the lung of SCID mice. In vitro study revealed elevated expression of stem cell associated markers like Oct4, Sox2 and Nanog as well as demonstrated intrinsic epithelial to mesenchymal transition features in SP cells. Further, we show that abrogation of EGFR, Src and Akt signaling through pharmacological or genetic inhibitors suppresses the self-renewal growth and expansion of SP-cells and resulted in specific downregulation of Sox2 protein expression. siRNA mediated depletion of Sox2 significantly blocked the SP phenotype as well as its self-renewal capacity; whereas other transcription factors like Oct4 and Nanog played a relatively lesser role in regulating self-renewal. Interestingly, Sox2 was elevated in metastatic foci of human NSCLC samples. Conclusions Our findings suggest that Sox2 is a novel target of EGFR-Src-Akt signaling in NSCLCs that modulates self-renewal and expansion of

Golden Trail: Retrieving the Data History that Matters from a Comprehensive Provenance Repository

Directory of Open Access Journals (Sweden)

Paolo Missier

2012-03-01

Full Text Available Experimental science can be thought of as the exploration of a large research space, in search of a few valuable results. While it is this “Golden Data” that gets published, the history of the exploration is often as valuable to the scientists as some of its outcomes. We envision an e-research infrastructure that is capable of systematically and automatically recording such history – an assumption that holds today for a number of workflow management systems routinely used in e-science. In keeping with our gold rush metaphor, the provenance of a valuable result is a “Golden Trail”. Logically, this represents a detailed account of how the Golden Data was arrived at, and technically it is a sub-graph in the much larger graph of provenance traces that collectively tell the story of the entire research (or of some of it.In this paper we describe a model and architecture for a repository dedicated to storing provenance traces and selectively retrieving Golden Trails from it. As traces from multiple experiments over long periods of time are accommodated, the trails may be sub-graphs of one trace, or they may be the logical representation of a virtual experiment obtained by joining together traces that share common data.The project has been carried out within the Provenance Working Group of the Data Observation Network for Earth (DataONE NSF project. Ultimately, our longer-term plan is to integrate the provenance repository into the data preservation architecture currently being developed by DataONE.

An evaluation of herbicides for post-emergence use in short rotation coppice

Energy Technology Data Exchange (ETDEWEB)

Turnbull, D.J.

2000-07-01

The objective of the project was to evaluate the safety and efficacy of a range of herbicides and mixtures of herbicides, with both contact and residual activity, for the post-emergence control of weeds in newly planted willow short rotation coppice (SRC). This report provides growers and advisers of short rotation coppice with important (but still limited) information on how to achieve improved weed control of problem weeds increasingly prevalent in SRC fields. This may provide guidance towards often-essential emergency treatments when the crop establishment is under considerable pressure and the potential safety, or otherwise, of certain weed-specific herbicides. (author)

Variability in growth of Vachellia nilotica provenances tested in the Sudano-Sahelian zone of Niger

DEFF Research Database (Denmark)

Larwanou, Mahamane; Issa, Rabiou; Saadou, Mahamane

2014-01-01

A provenance trial of Vachellia nilotica (Acacia nilotica) was conducted in Niger in order to assess variability in growth among 10 provenances from Africa (subsp. adstringens from Niger, Senegal and Cameroun) and outside the continent (subsp. indica from Pakistan and Yemen). Tree height, diameter...... at breast height and crown diameter were measured 15 years after trial establishment. Comparison of blocks located at slightly different elevation showed that trees had better survival and growth at the lower sites. The African provenances had better survival and total basal area than provenances from Yemen...

Features of Scots pine radial growth in conditions of provenance trial.

Science.gov (United States)

Kuzmin, Sergey; Kuzmina, Nina

2013-04-01

Provenance trial of Scots pine in Boguchany forestry of Krasnoyarsk krai is conducted on two different soils - dark-grey loam forest soil and sod-podzol sandy soil. Complex of negative factors for plant growth and development appears in dry conditions of sandy soil. It could results in decrease of resistance to diseases. Sandy soils in different climatic zones have such common traits as low absorbing capacity, poorness of elemental nutrition, low microbiological activity and moisture capacity, very high water permeability. But Scots pine trees growing in such conditions could have certain advantages and perspectives of use. In the scope of climate change (global warming) the study of Scots pine growth on sandy soil become urgent because of more frequent appearance of dry seasons. Purpose of the work is revelation of radial growth features of Scots pine with different origin in dry conditions of sandy soil and assessment of external factors influence. The main feature of radial growth of majority of studied pine provenances in conditions of sandy soil is presence of significant variation of increment with distinct decline in 25-years old with loss of tree rings in a number of cases. The reason of it is complex of factors: deficit of June precipitation and next following outbreak of fungal disease. Found «frost rings» for all trees of studied clymatypes in 1992 are the consequence of temperature decline from May 21 to June 2 - from 23 down to 2 degree Celsius. Perspective climatypes with biggest radial increments and least sensitivity to fungal disease were revealed. Eniseysk and Vikhorevka (from Krasnoyarsk krai and Irkutsk oblast)provenances of pine have the biggest radial increments, the least sensitivity to Cenangium dieback and smallest increments decline. These climatypes are in the group of perspective provenances and in present time they are recommended for wide trial in the region for future use in plantation forest growing. Kandalaksha (Murmansk oblast