Sample records for residual proven src

  1. Residue-specific description of non-native transient structures in the ensemble of acid-denatured structures of the all-beta protein c-src SH3

    Rösner, Heike I; Poulsen, Flemming Martin


    Secondary chemical shift analysis has been used to characterize the unfolded state of acid-denatured c-src SH3. Even though native c-src SH3 adopts an all-beta fold, we found evidence of transient helicity in regions corresponding to native loops. In particular, residues 40-46, connecting the n......-src loop to the third beta-strand, exhibited an apparent helicity of nearly 45%. Furthermore, the RT loop and the diverging turn appeared to adopt non-native-like helical conformations. Interestingly, none of the residues found in transient helical conformations exhibited significant varphi-values [Riddle......, D. S., et al. (1999) Nat. Struct. Biol. 6, 1016-1024]. This indicated that the transient helicity has no influence or only a weak influence on the actual protein folding reaction. The residual structural propensities were compared to those of other SH3 domains, revealing heterogeneity...

  2. Provenance Traces

    Cheney, James; Ahmed, Amal


    Provenance is information about the origin, derivation, ownership, or history of an object. It has recently been studied extensively in scientific databases and other settings due to its importance in helping scientists judge data validity, quality and integrity. However, most models of provenance have been stated as ad hoc definitions motivated by informal concepts such as "comes from", "influences", "produces", or "depends on". These models lack clear formalizations describing in what sense the definitions capture these intuitive concepts. This makes it difficult to compare approaches, evaluate their effectiveness, or argue about their validity. We introduce provenance traces, a general form of provenance for the nested relational calculus (NRC), a core database query language. Provenance traces can be thought of as concrete data structures representing the operational semantics derivation of a computation; they are related to the traces that have been used in self-adjusting computation, but differ in impor...

  3. Effects of Src on Proliferation and Invasion of Lung Cancer Cells

    Rui ZHENG


    Full Text Available Background and objective It has been proven that Src played pivotal roles in carcinogenesis, cancer progression and metastasis. The aim of this study is to explore the roles of Src phosphorylation on lung cancer cells. Methods Western blot and immunoprecipitation was used to detect the expression and phosphorylation of Src in lung cancer cells. MTT and Boyden chamber assay was used to examine the effects of inhibition of Src phosphorylation on proliferation and invasion of lung cancer cells in vitro, respectively. Results pp60src was expressed in all lung cancer cell lines in this study. All 5 non-small cell lung cancer (NSCLC cell lines had increased autophosphorylated tyrosine-418, while nearly no phosphorylated Src in small cell lung cancer SBC5 cell line was detected. The effect of inhibition of Src tyrosine kinase on cell proliferation varied among the lung cancer cell lines. Submicromolar Src tyrosine kinase inhibitor (≤1 μM remarkably suppressed the proliferation of PC-9 and A549 cells in a dose dependent manner (P < 0.05, while the same concentration of Src tyrosine kinase inhibitor had no significant effect on proliferation of H226, PC14PE6 and RERFLCOK cells. Invasiveness of lung cancer cells was significantly suppressed by Src tyrosine kinase in a dose-dependent manner (P < 0.05. Conclusion Phosphorylation of Src, but not over-expression, plays a pivotal role in proliferation and invasion of NSCLC cell lines in vitro.

  4. The SRC-II process

    Schmid, B. K.; Jackson, D. M.


    The Solvent Refined Coal (SRC-II) process which produces low-sulfur distillate fuel oil from coal is discussed. The process dissolves coal in a process-derived solvent at elevated temperature and pressure in the presence of hydrogen, separates the undissolved mineral residue, then recovers the original solvent by vacuum distillation. The distillate fuel oil produced is for use largely as a nonpolluting fuel for generating electrical power and steam and is expected to be competitive with petroleum fuels during the 1980s. During this period, the SRC-II fuel oil is expected to be attractive compared with combustion of coal with flue gas desulfurization in U.S. East Coast oil-burning power plants, as well as in small and medium-sized industrial boilers. The substantial quantities of methane, light hydrocarbons and naphtha produced by the process have value as feedstocks for preparation of pipeline gas, ethylene and high-octane unleaded gasoline, and can replace petroleum fractions in many applications. The liquid and gas products from a future large-scale plant, such as the 6000 t/day plant planned for Morgantown, West Virginia, are expected to have an overall selling price of $4.25 to $4.75/GJ.

  5. Identification of c-Src tyrosine kinase substrates using mass spectrometry and peptide microarrays

    Amanchy, Ramars; Zhong, Jun; Molina, Henrik;


    c-Src tyrosine kinase plays a critical role in signal transduction downstream of growth factor receptors, integrins and G protein-coupled receptors. We used stable isotope labeling with amino acids in cell culture (SILAC) approach to identify additional substrates of c-Src tyrosine kinase in human...... embryonic kidney 293T cells. We have identified 10 known substrates and interactors of c-Src and Src family kinases along with 26 novel substrates. We have experimentally validated 4 of the novel proteins (NICE-4, RNA binding motif 10, FUSE-binding protein 1 and TRK-fused gene) as direct substrates of c......-Src using in vitro kinase assays and cotransfection experiments. Significantly, using a c-Src specific inhibitor, we were also able to implicate 3 novel substrates (RNA binding motif 10, EWS1 and Bcl-2 associated transcription factor) in PDGF signaling. Finally, to identify the exact tyrosine residues...

  6. Fertilization of SRC willow. I

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


    Short rotation coppice (SRC) willow is often regarded as one of the most promising crops to increase biomass production and thereby meet the growing demand for renewable energy. This study is based on the hypotheses that biomass production of SRC willow responds positively to increasing doses...

  7. Environmental Stress Testing of the Single Sample Cylinder: A Proven Consensus Standard for Internal Gas Analysis (IGA) or Residual Gas Analysis (RGA)

    Schuessler, Philipp WH


    In August 2008, Schuessler Consulting was contracted by NASA GSFC in support of the NASA Electronic Parts and Packaging (NEPP) program to perform two separate studies on moisture laden air in a stainless steel cylinder that had been designed to become a consensus standard for Test Method 1018. This Test Method was originally released for hybrids under Mil. Std. 883 but was quickly utilized on other microelectronic devices under the auspice of Mil. Std. 750. The cylinder had subsequently been fabricated for the 750 community. It was back-filled with moist air and subsequently analyzed over a period of time under a previous NASA contract. It had been shown that moisture in the 4000 - 5000 ppm range could be analyzed rather precisely with a mass spectrometer, commonly referred to as a Residual Gas Analyzer (RGA). The scope of this study was to ascertain if the composition and precision varied as a function of thermal shock at sub-zero temperatures and whether there was consensus when the standard was submitted to other RGA units. It was demonstrated and published that the consensus standard would yield precise RGA data for moisture within +/- 1% when optimized for a given RGA unit. It has been subsequently shown in this study at Oneida Research Services, that sub-zero storage did not affect that precision when a well-defined protocol for the analysis was followed. The consensus standard was taken to a second facility for analysis where it was found that moisture adsorption on the transfer lines caused precision to drop to +/- 12%. The Single Sample Cylinder (SSC) is a one liter stainless steel cylinder with associated sampling valves and has considerable weight and volume. But this considerable size allows for approximately 300 gas samples of the same composition to be delivered to any RGA unit. Lastly, a smaller cylinder, approximately 75 cc, of a second consensus standard was fabricated and tested with a different mix of fixed gases where moisture was kept in the

  8. Dynamin Reduces Pyk2 Y402 Phosphorylation and Src Binding in Osteoclasts ▿ †

    Bruzzaniti, Angela; Neff, Lynn; Sandoval, Amanda; Du, Liping; Horne, William C.; Baron, Roland


    Signaling via the Pyk2-Src-Cbl complex downstream of integrins contributes to the assembly, organization, and dynamics of podosomes, which are the transient adhesion complexes of highly motile cells such as osteoclasts and dendritic cells. We previously demonstrated that the GTPase dynamin is associated with podosomes, regulates actin flux in podosomes, and promotes bone resorption by osteoclasts. We report here that dynamin associates with Pyk2, independent of dynamin's GTPase activity, and reduces Pyk2 Y402 phosphorylation in a GTPase-dependent manner, leading to decreased Src binding to Pyk2. Overexpressing dynamin decreased the macrophage colony-stimulating factor- and adhesion-induced phosphorylation of Pyk2 in osteoclastlike cells, suggesting that dynamin is likely to regulate Src-Pyk2 binding downstream of integrins and growth factor receptors with important cellular consequences. Furthermore, catalytically active Src promotes dynamin-Pyk2 association, and mutating specific Src-phosphorylated tyrosine residues in dynamin blunts the dynamin-induced decrease in Pyk2 phosphorylation. Thus, since Src binds to Pyk2 through its interaction with phospho-Y402, our results suggest that Src activates a negative-feedback loop downstream of integrin engagement and other stimuli by promoting both the binding of dynamin to Pyk2-containing complexes and the dynamin-dependent decrease in Pyk2 Y402 phosphorylation, ultimately leading to the dissociation of Src from Pyk2. PMID:19380485

  9. Dynamin reduces Pyk2 Y402 phosphorylation and SRC binding in osteoclasts.

    Bruzzaniti, Angela; Neff, Lynn; Sandoval, Amanda; Du, Liping; Horne, William C; Baron, Roland


    Signaling via the Pyk2-Src-Cbl complex downstream of integrins contributes to the assembly, organization, and dynamics of podosomes, which are the transient adhesion complexes of highly motile cells such as osteoclasts and dendritic cells. We previously demonstrated that the GTPase dynamin is associated with podosomes, regulates actin flux in podosomes, and promotes bone resorption by osteoclasts. We report here that dynamin associates with Pyk2, independent of dynamin's GTPase activity, and reduces Pyk2 Y402 phosphorylation in a GTPase-dependent manner, leading to decreased Src binding to Pyk2. Overexpressing dynamin decreased the macrophage colony-stimulating factor- and adhesion-induced phosphorylation of Pyk2 in osteoclastlike cells, suggesting that dynamin is likely to regulate Src-Pyk2 binding downstream of integrins and growth factor receptors with important cellular consequences. Furthermore, catalytically active Src promotes dynamin-Pyk2 association, and mutating specific Src-phosphorylated tyrosine residues in dynamin blunts the dynamin-induced decrease in Pyk2 phosphorylation. Thus, since Src binds to Pyk2 through its interaction with phospho-Y402, our results suggest that Src activates a negative-feedback loop downstream of integrin engagement and other stimuli by promoting both the binding of dynamin to Pyk2-containing complexes and the dynamin-dependent decrease in Pyk2 Y402 phosphorylation, ultimately leading to the dissociation of Src from Pyk2.

  10. Ecological fate and effects of solvent-refined-coal (SRC) materials: a status report

    Strand, J.A. III; Vaughan, B.E. (eds.)


    Non-occupational health effects associated with SRC operation will be determined by environmental factors governing the form, transport, and persistence of SRC materials and wastes - factors which also mediate exposure to man. Accordingly, the research described is an attempt to determine the fate of disposed solid wastes and spilled SRC materials, and it necessarily focuses on water soluble, persistent materials with greatest potential for mobility and incorporation into water and food supplies. Initially, aqueous equilibrations of SRC-II liquid material and SRC-I nongasified mineral residue were subjected to chemical characterization. Subsequently, laboratory studies were performed on the interaction of aqueous equilibrates of SRC-II liquid and SRC-I non-gasified mineral residue with soil materials isolated suspended sediments, and bottom sediments. These studies were designed to identify effects of specific sorption reactions ion or induced-ion exchange reactions, and toxicity of water soluble, biologically active materials derived from liquid and solid wastes. Results of these experiments have applicability to the environmental fate and effects of biologically active compounds released under different scenarios from product spills and solid waste disposal.

  11. Activation of BTK by a Phosphorylation Mechanism Initiated by SRC Family Kinases

    Rawlings, David J.; Scharenberg, Andrew M.; Park, Hyunsun; Wahl, Matthew I.; Lin, Siqi; Kato, Roberta M.; Fluckiger, Anne-Catherine; Witte, Owen N.; Kinet, Jean-Pierre


    Bruton's tyrosine kinase (BTK) is pivotal in B cell activation and development through its participation in the signaling pathways of multiple hematopoietic receptors. The mechanisms controlling BTK activation were studied here by examination of the biochemical consequences of an interaction between BTK and SRC family kinases. This interaction of BTK with SRC kinases transphosphorylated BTK on tyrosine at residue 551, which led to BTK activation. BTK then autophosphorylated at a second site. The same two sites were phosphorylated upon B cell antigen receptor cross-linking. The activated BTK was pre-dominantly membrane-associated, which suggests that BTK integrates distinct receptor signals resulting in SRC kinase activation and BTK membrane targeting.

  12. Fertilization of SRC willow. II

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


    Short rotation coppice (SRC) willow is an emerging cropping system in focus for production of biomass for energy. To increase production, the willow is commonly fertilized, but studies have shown differing effects of fertilization on biomass production, ranging from almost no response...... 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....... “Fertilization of SRC Willow, I: Biomass Production Response” Bioenergy Research, 2013). Nitrogen leaching was generally low, between 1 and 7 kg N ha−1 year−1 when doses of up to 120 kg N ha−1 year−1 were applied. Higher doses of 240 and 360 kg N ha−1 as single applications caused leaching of 66 and 99 kg N ha−1...

  13. The importance of Src signaling in sarcoma

    Chen, Quanchi; Zhou, Zifei; Shan, Liancheng; Zeng, Hui; Hua, Yingqi; Cai, Zhengdong


    Src is a tyrosine kinase that is of significance in tumor biology. The present review focuses on Src, its molecular structure, and role in cancer, in addition to its expression and function in sarcoma. In addition, the feasibility of Src as a potential drug target for the treatment of sarcoma is also discussed. Previous studies have suggested that Src has essential functions in cell proliferation, apoptosis, invasion, metastasis and the tumor microenvironment. Thus, it may be a potential targ...

  14. Hydroprocessing SRC. Final technical report

    Bronfenbrenner, J.C.; Garg, D.; Harris, C.F.; Znaimer, S.


    Catalyst activity and aging rate were studied in ICRC's process development unit (PDU) and at the Wilsonville Advanced Coal Liquefaction Facility under SRC-I Demonstration Plant hydroprocessing conditions. Similar studies using both high- and low-conversion modes were conducted by The Lummus Company. The studies determined variations in SRC conversion, hydrocarbon gas production, hydrogen consumption, and heteroatom removal. Samples of spent catalyst were analyzed to ascertain the reasons for catalyst deactivation. Finally, the ICRC PDU hydroprocessing results were compared with those generated at Lummus and Wilsonville pilot plants.

  15. SRC-I quarterly technical report, July-September 1980


    Processes in the SRC-I Demonstration plant are considered with respect to engineering, alternatives and comparative evaluations as follows: hydrogen production by coal gasification and gasification of residues, optimium pressure of hydrogen produced, shift processes, solidification of solvent-refined coal, effect of variations in coal and degree of coal preparation on coal liquefaction, characterization of various waste waters and comparative evaluation of three waste water treatment processes (including zero discharge). Nine papers have been entered individually into EDB and ERA. (LTN)

  16. Connexin43 recruits PTEN and Csk to inhibit c-Src activity in glioma cells and astrocytes

    González-Sánchez, Ana; Jaraíz-Rodríguez, Myriam; Domínguez-Prieto, Marta; Herrero-González, Sandra; Medina, José M.; Tabernero, Arantxa


    Connexin43 (Cx43), the major protein forming gap junctions in astrocytes, is reduced in high-grade gliomas, where its ectopic expression exerts important effects, including the inhibition of the proto-oncogene tyrosine-protein kinase Src (c-Src). In this work we aimed to investigate the mechanism responsible for this effect. The inhibition of c-Src requires phosphorylation at tyrosine 527 mediated by C-terminal Src kinase (Csk) and dephosphorylation at tyrosine 416 mediated by phosphatases, such as phosphatase and tensin homolog (PTEN). Our results showed that the antiproliferative effect of Cx43 is reduced when Csk and PTEN are silenced in glioma cells, suggesting the involvement of both enzymes. Confocal microscopy and immunoprecipitation assays confirmed that Cx43, in addition to c-Src, binds to PTEN and Csk in glioma cells transfected with Cx43 and in astrocytes. Pull-down assays showed that region 266–283 in Cx43 is sufficient to recruit c-Src, PTEN and Csk and to inhibit the oncogenic activity of c-Src. As a result of c-Src inhibition, PTEN was increased with subsequent inactivation of Akt and reduction of proliferation of human glioblastoma stem cells. We conclude that the recruitment of Csk and PTEN to the region between residues 266 and 283 within the C-terminus of Cx43 leads to c-Src inhibition. PMID:27391443

  17. Provenance Tracking in UNICORE

    Giesler, André; Hagemeier, Björn; Czekala, Myriam


    The automated tracking and storage of provenance information allows users of scientific workflow systems to validate and reproduce results of their experiments. Until now, UNICORE has not been providing comprehensive provenance features. For this reason, we plan to equip UNICORE with a flexible provenance tracking mechanism. Our goal is to ensure a suitable traceability of job and workflow processes into a description format supporting query capabilities and interoperability. In a first step,...

  18. Flow-Based Provenance

    Sabah Al-Fedaghi


    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.

  19. Specific interactions outside the proline-rich core of two classes of Src homology 3 ligands.

    Feng, S; Kasahara, C; Rickles, R J; Schreiber, S L


    Two dodecapeptides belonging to distinct classes of Src homology 3 (SH3) ligands and selected from biased phage display libraries were used to investigate interactions between a specificity pocket in the Src SH3 domain and ligant residues flanking the proline-rich core. The solution structures of c-Src SH3 complexed with these peptides were solved by NMR. In addition to proline-rich, polyproline type II helix-forming core, the class I and II ligands each possesses a flanking sequence that occupies a large pocket between the RT and n-Src loops of the SH3 domain. Structural and mutational analyses illustrate how the two classes of SH3 ligands exploit a specificity pocket on the receptor differently to increase binding affinity and specificity. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 Fig. 5 Fig. 6 Fig. 7 PMID:8618911

  20. SRC kinase regulation in progressively invasive cancer.

    Weichen Xu

    Full Text Available Metastatic progression is a multistep process that involves tumor growth and survival, motility and invasion, and subsequent proliferation in an inappropriate environment. The Src protein tyrosine kinase has been implicated in many of the biochemical pathways that drive these behaviors. Although Src itself is only rarely mutated in human tumors, its aberrant activity has been noted in various cancers and suggested to serve as a barometer of metastatic potential. With these features in mind, we examined Src kinase regulation at the structural, enzymatic, and expression levels as a function of progressively invasive prostate cancer cell lines. Surprisingly, both total Src content and kinase activity decrease with increasing cell line aggressiveness, an observation that appears to be inconsistent with the well-documented role of Src in the signaling pathways that drive growth and invasion. However, we do observe a direct correlation between Src kinase specific activity (total Src kinase activity/total Src content and metastatic aggressiveness, possibly suggesting that in highly aggressive cell lines, key signaling enzymes are globally recruited to drive the cancerous phenotype. In addition, although the expected enhanced phosphorylation of Src at Tyr-416 (activation site is present in the most aggressive prostate cancer cell lines, unexpectedly high phosphorylation levels at the Tyr-527 inhibitory site are observed as well. The latter, rather than representative of inhibited enzyme, is more indicative of primed Src responsive to local phosphorylated binding partners.

  1. SRC burn test in 700-hp oil-designed boiler. Annex Volume B. DOE-Pittsburgh Energy Technology Center report. Final technical report


    Solvent Refined Coal (SRC) combustion tests were conducted at the U.S. Department of Energy's Pittsburgh Energy Technology Center. Combustion and flue-gas treatment of three different physical forms of SRC, as well as a No. 6 fuel oil, were evaluated. The three SRC fuels were (1) pulverized SRC Fuel; (2) SRC Residual Fuel Oil; and (3) SRC/Water Slurry. The SRC Residual Fuel Oil was a solution of SRC Fuel dissolved in heated process solvent. Approximately 500 tons of pulverized SRC Fuel and 30,000 gallons of SRC Residual Fuel Oil were combusted in a 700 hp (30 x 130 x 10/sup 6/ Btu/hr fuel input) oil-designed watertube package boiler. Sixty four-hour ASME combustion tests with three different SRC fuels were successfully concluded. The principal parameters evaluated were excess air levels and combustion air preheat temperature levels. Extensive data were collected on flue-gas levels of O/sub 2/, CO/sub 2/, CO, unburned hydrocarbons, SO/sub x/, NO/sub x/, uncontrolled particulates, uncontrolled opacity and carbon content of the flue-gas particulates. Boiler and combustion efficiencies were measured. The particulates were characterized via mass loadings, impactors, in-situ resistivity measurements, ultra-fine sampling, optical large particle sampling, five-stage cyclone sampling and chemical analysis of various cut sizes. A three-field pilot electrostatic precipitator (ESP) containing over 1000 square feet of plate collection area, a reverse air fabric filter pilot dust collector and a commercial pulse-jet fabric filter dust collector were operated at high collection efficiency. The results will be valuable in making recommendations for future tests and will provide a basis for conversion of industrial oil-fired boilers to SRC fuels. 11 references, 20 figures, 29 tables.

  2. SRC burn test in 700-hp oil-designed boiler. Volume 2. Engineering evaluation report. Final technical report. [Oil-fired boiler to solvent-refined coal


    Volume 2 of this report gives the results of an engineering evaluation study and economic analysis of converting an existing 560-MW residual (No. 6) oil-fired unit to burn solvent refined coal (SRC) fuel forms. Volume 1 represents an integrated overview of the test program conducted at the Pittsburgh Energy Technology Center. Three SRC forms (pulverized SRC, a solution of SRC dissolved in process-derived distillates, and a slurry of SRC and water) were examined. The scope of modifications necessary to convert the unit to each of the three SRC fuel forms was identified and a capital cost of the necessary modifications estimated. A fuel conversion feasibility study of the boiler was performed wherein boiler modifications and performance effects of each fuel on the boiler were identified. An economic analysis of the capital and operating fuel expenses of conversion of the unit was performed. It was determined that conversion of the unit to any one of the three SRC fuel forms was feasible where appropriate modifications were made. It also was determined that the conversion of the unit can be economically attractive if SRC fuel forms can be manufactured and sold at prices discounted somewhat from the price of No. 16 Fuel Oil. As expected, greater discounts are required for the pulverized SRC and the slurry than for the solution of SRC dissolved in process-derived distillates.

  3. The Properties of Sulfur Rubber Concrete (SRC)


    The mix designs and specimen preparation for the dry process and wet process of sulfur rubber concrete (SRC) were investigated.The compressive strength, corrosion-resistance and toughness were studied and discussed.The results show that SRC is corrosion-resistanct.Although the compressive strength of SRC decreases with increasing rubber content, the toughness increases instead.Adding micro-filler will improve the compressive strength of SRC. There is a threshold value for the sulfur content, at which the compressive strength and the work ability of SRC reach an optimum balanc e.The bond between rubber particles and surrounding sulfur is strong due to the vulcanization process that generates cross-links through S-C bonds.

  4. Convergence of eicosanoid and integrin biology: Role of Src in 12-LOX activation.

    Dilly, Ashok-Kumar; Tang, Keqin; Guo, Yande; Joshi, Sangeeta; Ekambaram, Prasanna; Maddipati, Krishna Rao; Cai, Yinlong; Tucker, Stephanie C; Honn, Kenneth V


    12-Lipoxygenase (12-LOX) metabolizes arachidonic acid to 12(S)-hydroxyeicosatetraenoic acid, or 12(S)-HETE, a proinflammatory bioactive lipid implicated in tumor angiogenesis, growth, and metastasis. The mechanisms underlying 12-LOX-mediated signaling in cancer progression are still ill-defined. In the present study we demonstrate that 12-LOX phosphorylation and subsequent enzymatic activity occurs after integrin β4 stimulation and Src kinase recruitment to the integrin subunit. Inhibition of Src activity by PP2 or Src dominant-negative mutants reduced 12-LOX tyrosine phosphorylation and 12(S)-HETE production in response to integrin β4 stimulation in A431 cells. The pertinent Src-targeted residues for 12-LOX activity were mapped to Y19 and Y614, where 12-LOX mutants Y19F and Y614F showed 70% less enzymatic activity. Furthermore, we have shown that the 12-LOX activity modulated by these residues impacts migration. To our knowledge, this is the first report that c-Src kinase activity is required for β4-integrin-mediated phosphorylation of 12-LOX.

  5. Innocent Until Proven Guilty

    Case, Catherine; Whitaker, Douglas


    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…

  6. Proven Weight Loss Methods

    Fact Sheet Proven Weight Loss Methods What can weight loss do for you? Losing weight can improve your health in a number of ways. ... limiting calories) usually isn’t enough to cause weight loss. But exercise plays an important part in helping ...

  7. P-glycoprotein attenuates DNA repair activity in multidrug-resistant cells by acting through the Cbp-Csk-Src cascade.

    Lin, Li-Fang; Wu, Ming-Hsi; Pidugu, Vijaya Kumar; Ho, I-Ching; Su, Tsann-Long; Lee, Te-Chang


    Recent studies have demonstrated that P-glycoprotein (P-gp) expression impairs DNA interstrand cross-linking agent-induced DNA repair efficiency in multidrug-resistant (MDR) cells. To date, the detailed molecular mechanisms underlying how P-gp interferes with Src activation and subsequent DNA repair activity remain unclear. In this study, we determined that the C-terminal Src kinase-binding protein (Cbp) signaling pathway involved in the negative control of Src activation is enhanced in MDR cells. We also demonstrated that cells that ectopically express P-gp exhibit reduced activation of DNA damage response regulators, such as ATM, Chk2, Braca1 and Nbs1 and hence attenuated DNA double-strand break repair capacity and become more susceptible than vector control cells to DNA interstrand cross-linking (ICL) agents. Moreover, we demonstrated that P-gp can not only interact with Cbp and Src but also enhance the formation of inhibitory C-terminal Src kinase (Csk)-Cbp complexes that reduce phosphorylation of the Src activation residue Y416 and increase phosphorylation of the Src negative regulatory residue Y527. Notably, suppression of Cbp expression in MDR cells restores cisplatin-induced Src activation, improves DNA repair capacity, and increases resistance to ICL agents. Ectopic expression of Cbp attenuates cisplatin-induced Src activation and increases the susceptibility of cells to ICL agents. Together, the current results indicate that P-gp inhibits DNA repair activity by modulating Src activation via Cbp-Csk-Src cascade. These results suggest that DNA ICL agents are likely to have therapeutic potential against MDR cells with P-gp-overexpression.

  8. Data Provenance and Trust

    Stratis D Viglas


    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’.

  9. Provenance Store Evaluation

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


    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

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

    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.


    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).

  11. GRIM-19 inhibits v-Src-induced cell motility by interfering with cytoskeletal restructuring

    Sun, Peng; Nallar, Shreeram C.; Kalakonda, Sudhakar; Lindner, Daniel J.; Martin, Stuart S.; Kalvakolanu, Dhananjaya V.


    GRIM-19 (Gene associated with Retinoid-Interferon-induced Mortality 19) is a novel tumor suppressor regulated by Interferon/retinoid combination. We have recently shown that GRIM-19 inhibits v-Src-induced oncogenic transformation and metastatic behavior of cells. Oncogenic v-Src induces cell motility by cytoskeletal remodeling especially the formation of podosomes and. Here we show that GRIM-19 inhibited the v-Src-induced cell motility by inhibiting cytoskeletal remodeling i.e., podosome formation. We also show that the N-terminus of GRIM-19 played a major role in this process and identified critical residues in this region. More importantly, we show that tumor-associated GRIM-19 mutations disrupted its ability to inhibit v-Src-induced cell motility. These actions appear to occur independently of STAT3, a known target of GRIM-19-mediated inhibition. Lastly, tumor-associated GRIM-19 mutants significantly lost their ability to control v-Src-induced metastases in vivo, indicating the biological and pathological significance of these observations. PMID:19151760

  12. The Src family kinases: distinct functions of c-Src, Yes, and Fyn in the liver.

    Reinehr, Roland; Sommerfeld, Annika; Häussinger, Dieter


    The Src family kinases Yes, Fyn, and c-Src play a pivotal role in regulating diverse liver functions such as bile flow, proteolysis, apoptosis, and proliferation and are regulated by anisoosmotic cell volume changes, death receptor ligands, and bile acids. For example, cell swelling leads to an integrin-sensed and focal adhesion kinase-mediated activation of c-Src-triggering choleresis, proteolysis inhibition, regulatory volume decrease via p38MAPK and proliferation via the activation of the epidermal growth factor receptor and extracellular regulated kinases 1 and 2. In contrast, hepatocyte shrinkage generates an almost instantaneous oxidative stress response that triggers the activation of c-Jun N-terminal kinase and the Src family kinases Fyn and Yes. Whereas Fyn activation mediates cholestasis, Yes triggers CD95 activation and apoptosis. This review will discuss the role of Src family kinases in the regulation of liver function with emphasis on their role in osmo-signaling and bile acid signaling.

  13. Sewage sludge and wastewater fertilisation of Short Rotation Coppice (SRC) for increased bioenergy production - Biological and economic potential

    Dimitriou, I. [Department of Crop Production Ecology, Swedish University of Agricultural Sciences, P.O. Box 7043, SE 750 07 Uppsala (Sweden); Rosenqvist, H. [Department of Agriculture-Farming Systems, Technology and Product Quality, Swedish University of Agricultural Sciences, P.O. Box 17, SE-261 21 Billeberga (Sweden)


    Application of municipal residues, e.g. wastewater or sewage sludge, to Short Rotation Coppice (SRC) is among the most attractive methods for attaining environmental and energy goals set for Europe. At current woodchip prices in Sweden, the gross margin for SRC cultivation is positive only if biomass production is >9 t DM/ha yr. The gross profit margin increases (by 39 and 199 EUR/GJ, respectively) if sewage sludge and wastewater are applied to SRC. Application of residues to SRC has proved to be an acceptable alternative treatment method, and the farmer's profit can be markedly increased if compensation is paid for waste treatment. If all available sludge and wastewater were applied to SRC plantations, they could be grown on large agricultural areas in Europe, and c. 6000 PJ of renewable energy could be produced annually. However, a more economical landuse strategy, e.g. the use of more P-rich residues, appears more rational, and is biologically justifiable. (author)

  14. Difluoro analogue of UCS15A triggers activation of exogenously expressed c-Src in HCT 116 human colorectal carcinoma cells.

    Atatreh, Noor; Barraclough, Jane; Welman, Arkadiusz; Cawthorne, Christopher; Bryce, Richard A; Dive, Caroline; Freeman, Sally


    UCS 15A, an antibiotic produced by Streptomyces sp., has been reported to specifically disrupt SH3 domain-mediated interactions in eukaryotic cells. Interestingly, in the case of the non-receptor tyrosine kinase Src, UCS15A was effective in suppressing the SH3 domain-mediated intermolecular rather than intramolecular interactions, and thus prevented Src interactions with certain downstream effectors without affecting Src kinase activity. Here the synthesis of a novel difluoro analogue of UCS15A is described. The effects of this compound (8) on Src activity were tested in HCT 116 colorectal carcinoma cells engineered for inducible expression of c-Src. The presence of compound (8) resulted in the increased activity of the induced c-Src implicating that (8) acts as a c-Src activator in vivo. These observations are supported by computer modelling studies which suggest that the aldehyde group of (8) may covalently bind to a lysine residue in the SH2-kinase linker region situated in the proximity of the SH3 domain, which could promote a conformational change resulting in increased Src activity.

  15. Aggregation by Provenance Types: A Technique for Summarising Provenance Graphs

    Luc Moreau


    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.

  16. A switch role of Src in the biphasic EGF signaling of ER-negative breast cancer cells.

    XinTian Zhang

    Full Text Available It is well established that epidermal growth factor (EGF is a potent mitogen in cells expressing EGF receptor (EGFR. However, a body of evidence indicated that the effects of mitogenic EGF signaling exhibit a non-monotonic, or biphasic dose response curve; EGF at low concentrations elicits a mitogenic signaling pathway to stimulate cell proliferation while at high concentrations, EGF inhibits cell growth. However, the molecular mechanism underlying this paradoxical effect of EGF on cell proliferation remains largely unknown. Here, we investigated the molecular mechanisms underlying the biphasic EGF signaling in ER-negative breast cancer MDA-MB-231 and MDA-MB-436 cells, both of which express endogenous EGFR. We found that EGF at low concentrations induced the phosphorylation of the Src-Y416 residue, an event to activate Src, while at high concentrations allowed Src-Y527 phosphorylation that inactivates Src. EGF at 10 ng/ml also induced phosphorylation of the MAPK/ERK and activated cyclin D1 promoter activity through the Src/EGFR/STAT5 pathways but not at a higher concentration (500 ng/ml. Our results thus demonstrated that Src functions as a switch of EGF signaling depending on concentrations of EGF.

  17. Erratum to: Fertilization of SRC Willow

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


    Short rotation coppice (SRC) willow is an emerging cropping system in focus for production of biomass for energy. To increase production, the willow is commonly fertilized, but studies have shown differing effects of fertilization on biomass production, ranging from almost no response...... 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....... “Fertilization of SRC Willow, I: Biomass Production Response” Bioenergy Research, 2013). Nitrogen leaching was generally low, between 1 and 7 kg N ha−1 year−1 when doses of up to 120 kg N ha−1 year−1 were applied. Higher doses of 240 and 360 kg N ha−1 as single applications caused leaching of 66 and 99 kg N ha−1...

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

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


    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

  19. 76 FR 57763 - Alaska Region's Subsistence Resource Commission (SRC) Program


    ... Subsistence Resource Commission (SRC) program. SUMMARY: The Gates of the Arctic National Park SRC will meet to... locations and dates may need to be changed based on inclement weather or exceptional circumstances. Gates of the Arctic National Park SRC Meeting Dates and Location: The Gates of the Arctic National Park...

  20. Empowering Provenance in Data Integration

    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.

  1. Information Provenance in Social Media

    Barbier, Geoffrey; Liu, Huan

    Information appearing in social media provides a challenge for determining the provenance of the information. However, the same characteristics that make the social media environment challenging provide unique and untapped opportunities for solving the information provenance problem for social media. Current approaches for tracking provenance information do not scale for social media and consequently there is a gap in provenance methodologies and technologies providing exciting research opportunities for computer scientists and sociologists. This paper introduces a theoretical approach aimed guiding future efforts to realize a provenance capability for social media that is not available today. The guiding vision is the use of social media information itself to realize a useful amount provenance data for information in social media.

  2. Iron depletion results in Src kinase inhibition with associated cell cycle arrest in neuroblastoma cells.

    Siriwardana, Gamini; Seligman, Paul A


    Iron is required for cellular proliferation. Recently, using systematic time studies of neuroblastoma cell growth, we better defined the G1 arrest caused by iron chelation to a point in mid-G1, where cyclin E protein is present, but the cyclin E/CDK2 complex kinase activity is inhibited. In this study, we again used the neuroblastoma SKNSH cells lines to pinpoint the mechanism responsible for this G1 block. Initial studies showed in the presence of DFO, these cells have high levels of p27 and after reversal of iron chelation p27 is degraded allowing for CDK2 kinase activity. The initial activation of CDK2 kinase allows cells to exit G1 and enter S phase. Furthermore, we found that inhibition of p27 degradation by DFO is directly associated with inhibition of Src kinase activity measured by lack of phosphorylation of Src at the 416 residue. Activation of Src kinase occurs very early after reversal from the DFO G1 block and is temporally associated with initiation of cellular proliferation associated with entry into S phase. For the first time therefore we show that iron chelation inhibits Src kinase activity and this activity is a requirement for cellular proliferation.

  3. IVOA Provenance data model: hints from the CTA Provenance prototype

    Sanguillon, Michèle; Louys, Mireille; Bonnarel, François; Boisson, Catherine; Brégeon, Johan


    We present the last developments on the IVOA Provenance data model, mainly based on the W3C PROV concept. In the context of the Cherenkov astronomy, the data processing stages imply both assumptions and comparison to dedicated simulations. As a consequence, Provenance information is crucial to the end user in order to interpret the high level data products. The Cherenkov Telescope Array (CTA), currently in preparation, is thus a perfect test case for the development of an IVOA standard on Provenance information. We describe general use-cases for the computational Provenance in the CTA production pipeline and explore the proposed W3C notations like PROV-N formats, as well as Provenance access solutions.

  4. Dasatinib inhibits c-src phosphorylation and prevents the proliferation of Triple-Negative Breast Cancer (TNBC) cells which overexpress Syndecan-Binding Protein (SDCBP)

    Lang, Rong-Gang; Li, Wei-Dong; Sun, Hui; Liu, Fang-Fang; Guo, Xiao-Jing; Gu, Feng; Fu, Li


    Triple negative breast cancer (TNBC) progresses rapidly but lacks effective targeted therapies. Our previous study showed that downregulating syndecan-binding protein (SDCBP) in TNBC inhibits the proliferation of TNBC cells. Dasatinib is a new small-molecule inhibitor of c-src phosphorylation. The aim of this study was to investigate if SDCBP is a potential marker to indicate whether a TNBC is suitable for dasatinib therapy. This study applied co-immunoprecipitation to identify the interaction between SDCBP and c-src in TNBC cell lines. In addition, immunohistochemistry was used to investigate SDCBP and tyrosine-419 phosphorylated c-src (p-c-src-Y419) expression in TNBC tissues. SDCBP-overexpressing MDA-MB-231 cells were then constructed to evaluate the effects of dasatinib on SDCBP-induced TNBC progression in vitro and tumor formation in nude mice. We found wild-type SDCBP interacted with c-src and promoted the phosphorylation of c-src; this phosphorylation was completely blocked by dasatinib. SDCBP lacking the PDZ domain had no such effect. Among the 52 consecutive random TNBC cases examined, the expression of SDCBP was consistent with that of p-c-src-Y419, and positively correlated with histological grading or Ki-67 levels. SDCBP overexpression significantly accelerated the proliferation and cell cycle progression of the TNBC cell line MDA-MB-231; these effects were prevented by dasatinib treatment. However, the subsequent inhibition of p27 expression partially restored the proliferation and viability of the TNBC cells. The results of this study suggest that SDCBP interacts with c-src, regulates G1/S in TNBC cells, and enhances tumor cell proliferation by promoting the tyrosine phosphorylation of c-src at residue 419. Dasatinib inhibits such phosphorylation and blocks SDCBP-induced cell cycle progression. Therefore, SDCBP might be an important marker for identifying TNBC cases that are suitable for dasatinib therapy. PMID:28141839

  5. Src-like-adaptor protein (SLAP) differentially regulates normal and oncogenic c-Kit signaling.

    Kazi, Julhash U; Agarwal, Shruti; Sun, Jianmin; Bracco, Enrico; Rönnstrand, Lars


    The Src-like-adaptor protein (SLAP) is an adaptor protein sharing considerable structural homology with Src. SLAP is expressed in a variety of cells and regulates receptor tyrosine kinase signaling by direct association. In this report, we show that SLAP associates with both wild-type and oncogenic c-Kit (c-Kit-D816V). The association involves the SLAP SH2 domain and receptor phosphotyrosine residues different from those mediating Src interaction. Association of SLAP triggers c-Kit ubiquitylation which, in turn, is followed by receptor degradation. Although SLAP depletion potentiates c-Kit downstream signaling by stabilizing the receptor, it remains non-functional in c-Kit-D816V signaling. Ligand-stimulated c-Kit or c-Kit-D816V did not alter membrane localization of SLAP. Interestingly oncogenic c-Kit-D816V, but not wild-type c-Kit, phosphorylates SLAP on residues Y120, Y258 and Y273. Physical interaction between c-Kit-D816V and SLAP is mandatory for the phosphorylation to take place. Although tyrosine-phosphorylated SLAP does not affect c-Kit-D816V signaling, mutation of these tyrosine sites to phenylalanine can restore SLAP activity. Taken together the data demonstrate that SLAP negatively regulates wild-type c-Kit signaling, but not its oncogenic counterpart, indicating a possible mechanism by which the oncogenic c-Kit bypasses the normal cellular negative feedback control.

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



    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.

  7. A Noisy 10GB Provenance Database

    Cheah, You-Wei; Plale, Beth; Kendall-Morwick, Joey; Leake, David; Ramakrishnan, Lavanya


    Provenance of scientific data is a key piece of the metadata record for the data's ongoing discovery and reuse. Provenance collection systems capture provenance on the fly, however, the protocol between application and provenance tool may not be reliable. Consequently, the provenance record can be partial, partitioned, and simply inaccurate. We use a workflow emulator that models faults to construct a large 10GB database of provenance that we know is noisy (that is, has errors). We discuss the process of generating the provenance database, and show early results on the kinds of provenance analysis enabled by the large provenance.

  8. Provenance an introduction to PROV

    Moreau, Luc


    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

  9. Final technical report: SRC burn test in 700-hp oil-designed boiler. Annex Volume D. Electrostatic precipitator mass train and operating data


    Solvent Refined Coal (SRC) is one of the viable replacement fuels for No. 6 fuel oil in industrial and utility boilers. The Department of Energy funded the International Coal Refining Company (ICRC) to develop and to demonstrate the use of SRC as a practical fuel. Phase II of the project was to burn the SRC fuels in a 700 hp package boiler and to collect emission data from which air pollution control devices could be specified. Wheelabrator-Frye, Inc., APC Division was contracted by ICRC to supply and operate a pilot electrostatic precipitator (ESP). Mass emission testing was performed by WFI Sciences. Particle size tests, particle resistivity, SO/sub x/ measurements, and particulate counting tests were conducted by Southern Research Institute (SoRI). This report is a source document covering the ESP operating data and mass emission data. The data obtained by SoRI is used by SoRI in their computer model to specify full scale design criteria. The testing was performed with four fuel types; No. 6 fuel oil, SRC fuel, SRC residual fuel oil, and SRC-water slurry. All fuels were precipitated quite easily resulting in emission rates below the NSPS standards.

  10. Chemical, biomedical and ecological studies of SRC-I materials from the Fort Lewis Pilot Plant: a status report

    Mahlum, D.D. (ed.)


    This document discusses studies performed with solvent refined coal (SRC) materials obtained from the Fort Lewis Pilot Plant during operation in the SRC-I mode. The development of analytical methodology is presented as well as results obtained from the application of these methods to light oil (LO), wash solvent (WS) and process solvent (PS). Results of cellular and animal studies with LO, WS and PS are included, along with a description of methods for the generation and characterization of LO and PS aerosols, and for exposing rats, mice and guinea pigs to these aerosols. The effects of SRC-I product on seed germination and plant growth which have also been studied are discussed. The SRC-I product, feed coal and the mineral residue have been analyzed for organic and inorganic constituents. The higher-boiling-point material, PS, exhibited significant mutagenic activity in the Ames assay; LO and WS were inactive. Process solvent also caused transformation of cultured Syrian hamster embryo cells. Additional chemical fractionation studies suggest that primary aromatic amines are major determinants of the observed mutagenic activity. Skin-painting studies with SRC-II naphtha, heavy distillate, shale oil and petroleum crude indicate a good correlation between the results of the cellular assays and skin carcinogenesis in mice. Wash solvent was more toxic after oral administration to rats than was light oil or process solvent. The effects of LO, WS and PS on development were studied after administration to pregnant rats. The tissue distribution of a number of components of PS was studied after oral administration of PS to rats. The effect of SRC-I product on the germination and growth of barley was investigated by mixing or layering the product with soil and placing the mixture in a field lysimeter.

  11. Provenance Data in Social Media


    provenance on the web. Foundations and Trends in Web Science, 2:99–241, 2009. DOI: 10.1561/1800000010. 23 [48] N. Naveed , T. Gottron, J. Kunegis, and A. C...Computer Science and Engineering, Arizona State University (ASU). He obtained his Ph.D. in the Department of Systems and Industrial Engineering at the

  12. 77 FR 4580 - Alaska Region's Subsistence Resource Commission (SRC) Program


    ... 19, 2011), was canceled due to a lack of quorum caused by inclement Arctic weather conditions. The... weather or exceptional circumstances. Kobuk Valley National Park SRC Meeting Date and Location: The Kobuk Valley National Park SRC will meet at the National Park Service Northwest Arctic Heritage Center,...

  13. Non-cysteine linked MUC1 cytoplasmic dimers are required for Src recruitment and ICAM-1 binding induced cell invasion

    Gunasekara Nirosha


    Full Text Available Abstract Background The mucin MUC1, a type I transmembrane glycoprotein, is overexpressed in breast cancer and has been correlated with increased metastasis. We were the first to report binding between MUC1 and Intercellular adhesion molecule-1 (ICAM-1, which is expressed on stromal and endothelial cells throughout the migratory tract of a metastasizing breast cancer cell. Subsequently, we found that MUC1/ICAM-1 binding results in pro-migratory calcium oscillations, cytoskeletal reorganization, and simulated transendothelial migration. These events were found to involve Src kinase, a non-receptor tyrosine kinase also implicated in breast cancer initiation and progression. Here, we further investigated the mechanism of MUC1/ICAM-1 signalling, focusing on the role of MUC1 dimerization in Src recruitment and pro-metastatic signalling. Methods To assay MUC1 dimerization, we used a chemical crosslinker which allowed for the detection of dimers on SDS-PAGE. We then generated MUC1 constructs containing an engineered domain which allowed for manipulation of dimerization status through the addition of ligands to the engineered domain. Following manipulation of dimerization, we immunoprecipitated MUC1 to investigate recruitment of Src, or assayed for our previously observed ICAM-1 binding induced events. To investigate the nature of MUC1 dimers, we used both non-reducing SDS-PAGE and generated a mutant construct lacking cysteine residues. Results We first demonstrate that the previously observed MUC1/ICAM-1signalling events are dependent on the activity of Src kinase. We then report that MUC1 forms constitutive cytoplasmic domain dimers which are necessary for Src recruitment, ICAM-1 induced calcium oscillations and simulated transendothelial migration. The dimers are not covalently linked constitutively or following ICAM-1 binding. In contrast to previously published reports, we found that membrane proximal cysteine residues were not involved in

  14. 78 FR 51207 - Kobuk Valley National Park Subsistence Resource Commission (SRC) and the Denali National Park SRC...


    ... National Park Service Kobuk Valley National Park Subsistence Resource Commission (SRC) and the Denali National Park SRC; Meetings AGENCY: National Park Service, Interior. ACTION: Meeting notice. SUMMARY: As required by the Federal Advisory Committee Act (Public Law 92-463, 86 Stat. 770), the National Park...

  15. COOH-terminal association of human smooth muscle calcium channel Cav1.2b with Src kinase protein binding domains: effect of nitrotyrosylation

    Minho Kang; Gracious R. Ross; Hamid I. Akbarali


    ...) and the effect of nitrotyrosylation. Cotransfection of human embryonic kidney (HEK)-293 cells with hCav1.2b and c-Src resulted in tyrosine phosphorylation of the calcium channel, which was prevented by nitration of tyrosine residues by peroxynitrite...

  16. COOH-terminal association of human smooth muscle calcium channel Ca^sub v^1.2b with Src kinase protein binding domains: effect of nitrotyrosylation

    Minho Kang; Gracious R Ross; Hamid I Akbarali


    ...) and the effect of nitrotyrosylation. Cotransfection of human embryonic kidney (HEK)-293 cells with hCa...1.2b and c-Src resulted in tyrosine phosphorylation of the calcium channel, which was prevented by nitration of tyrosine residues by peroxynitrite...

  17. COOH-terminal association of human smooth muscle calcium channel Ca(v)1.2b with Src kinase protein binding domains: effect of nitrotyrosylation

    Kang, Minho; Ross, Gracious R; Akbarali, Hamid I


    ...) and the effect of nitrotyrosylation. Cotransfection of human embryonic kidney (HEK)-293 cells with hCa(v)1.2b and c-Src resulted in tyrosine phosphorylation of the calcium channel, which was prevented by nitration of tyrosine residues by peroxynitrite...

  18. ADAM12 localizes with c-Src to actin-rich structures at the cell periphery and regulates Src kinase activity

    Stautz, Dorte; Sanjay, Archana; Hansen, Matilde Thye;


    to enhance Src kinase activity in response to external signals, such as integrin engagement. Thus, we suggest that activated c-Src binds, phosphorylates, and redistributes ADAM12-L to specific sites at the cell periphery, which may in turn promote signalling mechanisms regulating cellular processes...... partners and signalling proteins. We demonstrate here a c-Src-dependent redistribution of ADAM12-L from perinuclear areas to actin-rich Src-positive structures at the cell periphery, and identified two separate c-Src binding sites in the cytoplasmic tail of ADAM12-L that interact with the SH3 domain of c......-Src with different binding affinities. The association between ADAM12-L and c-Src is transient, but greatly stabilized when the c-Src kinase activity is disrupted. In agreement with this observation, kinase-active forms of c-Src induce ADAM12-L tyrosine phosphorylation. Interestingly, ADAM12-L was also found...

  19. Provenance management in Swift with implementation details.

    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)


    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.

  20. Logical provenance in data-oriented workflows?

    Ikeda, R.


    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.

  1. File level provenance tracking in CMS

    Jones, C.D.; Kowalkowski, J.; Paterno, M.; Sexton-Kennedy, L.; Tanenbaum, W.; /Fermilab; Riley, D.S.; /Cornell U., LEPP


    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 minimize the space used to store the provenance in the file and the tools that will be available to use the provenance.

  2. File Level Provenance Tracking in CMS

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


    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.

  3. File level provenance tracking in CMS

    Jones, C D; Kowalkowski, J; Paterno, M; Sexton-Kennedy, E; Tanenbaum, W [Fermi National Accelerator Laboratory, PO Box 500, Batavia IL 60510-5011 (United States); Riley, D S, E-mail: cdj@fnal.go [Wilson Laboratory, Cornell University, Ithaca NY 14853-8001 (United States)


    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.

  4. SRC-3/AIB1:transcriptional coactivator in oncogenesis

    Jun YAN; Sophia Y TSAI; Ming-jer TSAI


    Steroid receptor coactivator-3 (SRC-3,also known as NCoA3,AIB1,p/CIP,RAC3,ACTR,and TRAM1) ,localized on a frequently amplified region,20q12,has been associated with multiple cancers,including breast,gastric and prostate cancers.Although SRC-3 has been implicated as an oncogene,compelling evidence has only recently emerged implicating it as a causal factor in the genesis of human cancers.Here,we summarize recent evidence that indicates aberrant SRC-3 expression is important in hormone-sensitive and -insensitive human cancers.

  5. Provenance data in social media

    Barbier, Geoffrey; Gundecha, Pritam


    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

  6. Lipids Regulate Lck Protein Activity through Their Interactions with the Lck Src Homology 2 Domain.

    Sheng, Ren; Jung, Da-Jung; Silkov, Antonina; Kim, Hyunjin; Singaram, Indira; Wang, Zhi-Gang; Xin, Yao; Kim, Eui; Park, Mi-Jeong; Thiagarajan-Rosenkranz, Pallavi; Smrt, Sean; Honig, Barry; Baek, Kwanghee; Ryu, Sungho; Lorieau, Justin; Kim, You-Me; Cho, Wonhwa


    Lymphocyte-specific protein-tyrosine kinase (Lck) plays an essential role in T cell receptor (TCR) signaling and T cell development, but its activation mechanism is not fully understood. To explore the possibility that plasma membrane (PM) lipids control TCR signaling activities of Lck, we measured the membrane binding properties of its regulatory Src homology 2 (SH2) and Src homology 3 domains. The Lck SH2 domain binds anionic PM lipids with high affinity but with low specificity. Electrostatic potential calculation, NMR analysis, and mutational studies identified the lipid-binding site of the Lck SH2 domain that includes surface-exposed basic, aromatic, and hydrophobic residues but not the phospho-Tyr binding pocket. Mutation of lipid binding residues greatly reduced the interaction of Lck with the ζ chain in the activated TCR signaling complex and its overall TCR signaling activities. These results suggest that PM lipids, including phosphatidylinositol 4,5-bisphosphate and phosphatidylinositol 3,4,5-trisphosphate, modulate interaction of Lck with its binding partners in the TCR signaling complex and its TCR signaling activities in a spatiotemporally specific manner via its SH2 domain. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  7. The imaging performance of the SRC on Mars Express

    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.


    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.

  8. Differential gene regulation by the SRC family of coactivators

    HuaZhang; XiaYi; Xiaojingsun; NaYin; BinShi; HuijianWu; DanWang; GeWu; YongfengShang


    SRCs (steroid receptor coactivatorsl are required for nuclear receptor-mediated transcription and are also implicated in the transcription initiation by other transcription factors, such as STATs and NFKB. Despite phenotypic manifestations in gene knockout mice for SRC-1, GRIP1, and AIB1 of the SRC (Steroid Receptor Coactivator) family indicating their differential roles in animal physiology, there is no clear evidence, at the molecular level, to support a functional specificity for these proteins. We demonstrated in this report that two species of SRC coactivators, either as AIBI:GRIP1 or as AIBI:SRC-1 are recruited, possibly through heterodimerization, on the promoter of genes that contain a classical hormone responsive element (HRE). In contrast, on non-HRE-containing gene promoters, on which steroid receptors bind indirectly, either GRIP1 orSRC-1 is recruited as a monomer, depending on the cellular abundance of the protein. Typically, non-HRE-containing genes are early genes activated by steroid receptors, whereas HRE-containing genes are activated later. Our results also showed that SRC proteins contribute to the temporal regulation of gene transcription. In addition, our experiments revealed a positive correlation between AIB1/c-myc overexpression in ER+ breast carcinoma samples, suggesting a possible mechanism for AIB1/n breast cancer carcinogenesis.

  9. Inhibition of N1-Src kinase by a specific SH3 peptide ligand reveals a role for N1-Src in neurite elongation by L1-CAM

    Keenan, Sarah; Wetherill, Sarah J.; Ugbode, Christopher I.; Chawla, Sangeeta; Brackenbury, William J.; Evans, Gareth J. O.


    In the mammalian brain the ubiquitous tyrosine kinase, C-Src, undergoes splicing to insert short sequences in the SH3 domain to yield N1- and N2-Src. We and others have previously shown that the N-Srcs have altered substrate specificity and kinase activity compared to C-Src. However, the exact functions of the N-Srcs are unknown and it is likely that N-Src signalling events have been misattributed to C-Src because they cannot be distinguished by conventional Src inhibitors that target the kinase domain. By screening a peptide phage display library, we discovered a novel ligand (PDN1) that targets the unique SH3 domain of N1-Src and inhibits N1-Src in cells. In cultured neurons, PDN1 fused to a fluorescent protein inhibited neurite outgrowth, an effect that was mimicked by shRNA targeting the N1-Src microexon. PDN1 also inhibited L1-CAM-dependent neurite elongation in cerebellar granule neurons, a pathway previously shown to be disrupted in Src−/− mice. PDN1 therefore represents a novel tool for distinguishing the functions of N1-Src and C-Src in neurons and is a starting point for the development of a small molecule inhibitor of N1-Src. PMID:28220894

  10. Understanding collaborative studies through interoperable workflow provenance

    Altintas, I.; Anand, M.K.; Crawl, D.; Bowers, S.; Belloum, A.; Missier, P.; Ludäscher, B.; Goble, C.A.; Sloot, P.M.A.


    The provenance of a data product contains information about how the product was derived, and is crucial for enabling scientists to easily understand, reproduce, and verify scientific results. Currently, most provenance models are designed to capture the provenance related to a single run, and mostly

  11. BOREAS AFM-07 SRC Surface Meteorological Data

    Osborne, Heather; Hall, Forrest G. (Editor); Newcomer, Jeffrey A. (Editor); Young, Kim; Wittrock, Virginia; Shewchuck, Stan; Smith, David E. (Technical Monitor)


    The Saskatchewan Research Council (SRC) collected surface meteorological and radiation data from December 1993 until December 1996. The data set comprises Suite A (meteorological and energy balance measurements) and Suite B (diffuse solar and longwave measurements) components. Suite A measurements were taken at each of ten sites, and Suite B measurements were made at five of the Suite A sites. The data cover an approximate area of 500 km (North-South) by 1000 km (East-West) (a large portion of northern Manitoba and northern Saskatchewan). The measurement network was designed to provide researchers with a sufficient record of near-surface meteorological and radiation measurements. The data are provided in tabular ASCII files, and were collected by Aircraft Flux and Meteorology (AFM)-7. The surface meteorological and radiation data are available from the Earth Observing System Data and Information System (EOSDIS) Oak Ridge National Laboratory (ORNL) Distributed Active Archive Center (DAAC). The data files are available on a CD-ROM (see document number 20010000884).

  12. Characterization of an Engineered Src Kinase to Study Src Signaling and Biology

    Gentry, Leanna R.; Karginov, Andrei V.; Hahn, Klaus M.; Der, Channing J.


    Summary Pharmacologic inhibitors of protein kinases comprise the vast majority of approved signal transduction inhibitors for cancer treatment. An important facet of their clinical development is the identification of the key substrates critical for their driver role in cancer. One approach for substrate identification involves evaluating the phosphorylation events associated with stable expression of an activated protein kinase. Another involves genetic or pharmacologic inhibition of protein kinase expression or activity. However, both approaches are limited by the dynamic nature of signaling, complicating whether phosphorylation changes are primary or secondary activities of kinase function. We have developed rapamycin-regulated (RapR) protein kinases as molecular tools that allow for the study of spatiotemporal regulation of signaling. Here we describe the application of this technology to the Src tyrosine kinase and oncoprotein (RapR-Src). We describe how to achieve stable expression of this tool in cell lines and how to subsequently activate the tool and determine its function in signaling and morphology. PMID:26501909

  13. Activation of Src and Src-associated signaling pathways in relation to hypoxia in human cancer xenograft models.

    Pham, Nhu-An; Magalhaes, Joao M M M; Do, Trevor; Schwock, Joerg; Dhani, Neesha; Cao, Ping-Jiang; Hill, Richard P; Hedley, David W


    The hypoxic response in vitro involves alterations in signaling proteins, including Src, STAT3 and AKT that are considered to be broadly pro-survival. The involvement of these signaling proteins in the hypoxic microenviroments that occur in solid tumors was investigated by the use of multicolor fluorescence image analysis to colocalize signaling proteins and regions of hypoxia in 4 human tumor xenografts, pancreatic carcinoma BxPC3 and PANC1 and cervical squamous cell carcinoma ME180 and SiHa. Expression levels of total Src protein (mean intensity x labeled region fraction) were higher in hypoxic regions, identified using the nitroimidazole probe EF5, relative to non-EF5 regions in all 4 tumor models. This was associated with higher levels of phosphorylated (p-) Y419p-Src and its substrate Y861p-FAK in EF5 positive regions of BxPC3 tumors. This effect was also seen in tumor-bearing mice continuously breathing 7% oxygen for 3 hr which markedly increased the extent of EF5 positive labeling. In contrast, the hypoxia treatment resulted in a significant decrease in S727p-STAT3 in BxPC3 xenografts and suggested that STAT3 activity is responsive to acute hypoxia, whereas Src-FAK signaling is associated with predominantly chronically hypoxic EF5 positive regions. Src activity in both hypoxic and nonhypoxic BxPC3 tumor regions was suppressed when mice were treated with the Src inhibitor AZD0530 (25 mg/kg/day, 5 days), suggesting that both hypoxic and normoxic tumor regions are accessible to pharmacological Src inhibition. These results show that signaling pathways are responsive to tumor hypoxia in vivo, although the effects appear to differ between individual tumor types. Copyright (c) 2008 Wiley-Liss, Inc.

  14. Tyrosine Phosphorylation of Tau by the Src Family Kinases Lck and Fyn

    Bird Ian N


    Full Text Available Abstract Background Tau protein is the principal component of the neurofibrillary tangles found in Alzheimer's disease, where it is hyperphosphorylated on serine and threonine residues, and recently phosphotyrosine has been demonstrated. The Src-family kinase Fyn has been linked circumstantially to the pathology of Alzheimer's disease, and shown to phosphorylate Tyr18. Recently another Src-family kinase, Lck, has been identified as a genetic risk factor for this disease. Results In this study we show that Lck is a tau kinase. In vitro, comparison of Lck and Fyn showed that while both kinases phosphorylated Tyr18 preferentially, Lck phosphorylated other tyrosines somewhat better than Fyn. In co-transfected COS-7 cells, mutating any one of the five tyrosines in tau to phenylalanine reduced the apparent level of tau tyrosine phosphorylation to 25-40% of that given by wild-type tau. Consistent with this, tau mutants with only one remaining tyrosine gave poor phosphorylation; however, Tyr18 was phosphorylated better than the others. Conclusions Fyn and Lck have subtle differences in their properties as tau kinases, and the phosphorylation of tau is one mechanism by which the genetic risk associated with Lck might be expressed pathogenically.

  15. ATP-mediated transactivation of the epidermal growth factor receptor in airway epithelial cells involves DUOX1-dependent oxidation of Src and ADAM17.

    Derek Sham

    Full Text Available The respiratory epithelium is subject to continuous environmental stress and its responses to injury or infection are largely mediated by transactivation of the epidermal growth factor receptor (EGFR and downstream signaling cascades. Based on previous studies indicating involvement of ATP-dependent activation of the NADPH oxidase homolog DUOX1 in epithelial wound responses, the present studies were performed to elucidate the mechanisms by which DUOX1-derived H(2O(2 participates in ATP-dependent redox signaling and EGFR transactivation. ATP-mediated EGFR transactivation in airway epithelial cells was found to involve purinergic P2Y(2 receptor stimulation, and both ligand-dependent mechanisms as well as ligand-independent EGFR activation by the non-receptor tyrosine kinase Src. Activation of Src was also essential for ATP-dependent activation of the sheddase ADAM17, which is responsible for liberation and activation of EGFR ligands. Activation of P2Y(2R results in recruitment of Src and DUOX1 into a signaling complex, and transient siRNA silencing or stable shRNA transfection established a critical role for DUOX1 in ATP-dependent activation of Src, ADAM17, EGFR, and downstream wound responses. Using thiol-specific biotin labeling strategies, we determined that ATP-dependent EGFR transactivation was associated with DUOX1-dependent oxidation of cysteine residues within Src as well as ADAM17. In aggregate, our findings demonstrate that DUOX1 plays a central role in overall epithelial defense responses to infection or injury, by mediating oxidative activation of Src and ADAM17 in response to ATP-dependent P2Y(2R activation as a proximal step in EGFR transactivation and downstream signaling.

  16. Evidence for involvement of c-Src in the anti-apoptotic action of nitric oxide in serum-deprived RINm5F cells.

    Tejedo, J R; Ramírez, R; Cahuana, G M; Rincón, P; Sobrino, F; Bedoya, F J


    The mechanism by which nitric oxide (NO) protects from apoptosis is a matter of debate. We have shown previously that phosphorylation of tyrosine residues participates in the protection from apoptosis in insulin-producing RINm5F cells (Inorg. Chem. Commun. 3 (2000) 32). Since NO has been reported to activate the tyrosine kinase c-Src and this kinase is involved in the activation of protein kinase G (PKG) in some cell systems, we aimed at studying the contribution of c-Src and PKG systems in anti-apoptotic actions of NO in serum-deprived RINm5F cells. Here we report that exposure of serum-deprived cells to 10 microM DETA/NO results in protection from degradation of the anti-apoptotic protein Bcl-2, together with a reduction of cytochrome c release from mitochondria and caspase-3 inhibition. Studies with the inhibitors ODQ and KT-5823 revealed that these actions are dependent on both activation of guanylate cyclase and PKG. DETA/NO was also able to induce autophosphorylation and activation c-Src protein both in vivo and in vitro and active c-Src was able to induce tyrosine phosphorylation of Bcl-2 in vitro. The c-Src kinase inhibitor PP1 abrogated the actions of DETA/NO on cGMP formation, PKG activation, caspase activation, cytochrome c release from mitochondria, and Bcl-2 phosphorylation and degradation in serum-deprived cells. We thus propose that activation of c-Src is an early step in the chain of events that signal cGMP-dependent anti-apoptotic actions of NO in mitocohondria.

  17. SRC-GAZEL: a full-scale pilot project

    Schenkel, Y.; Grulois, C. [Centre de Recherches Agronomiques, Gembloux (Belgium); Martin, J.; Bourgois, F.; Jossart, J.-M. [Universite Catholique de Louvain (France); Meekers, E. [Facultes Universitaires Notre Dame de la Paix (France)


    In the context of the European agricultural crisis and of set-aside land, the biomass cultivation alternative for energy generation increasingly interests the agricultural world. The Short Rotation Coppice-GAZEL project studies the feasibility of generating electricity by the gasification of short rotation coppice of willows or poplars. It perfectly fits in the Belgian general energy plan: one purpose of this plan is to set up small electric generation units feeding directly the low-voltage grid in order to bring a flexible component to the massive production of nuclear power plants. The project consists of three major parts: production of short rotation coppice (SRC); mechanisation of the culture of SRC and conditioning of the wood products for gasification; and gasification and electricity production. In addition, socio-economic and environmental impacts will be analysed in detail. In this paper, we focus on the choices that have to be made regarding SRC harvesting and fuel conditioning. (author)

  18. Väike SRC Grupp ähvardab BLRT-le kandadele astuda / Agnes Ojala

    Ojala, Agnes


    SRC Grupp ja rahvusvaheline laevanduskontsern Aker Yard sõlmisid koostöölepingu, et pakkuda Läänemere põhjaregioonis tegutsevatele laevandusettevõtetele tipptasemel laevaremonditeenust. Diagramm: SRC ja BLRT käive

  19. SGProv: Summarization Mechanism for Multiple Provenance Graphs

    El-Jaick, Daniele; Mattoso, Marta; Lima, Alexandre A. B.


    Scientific workflow management systems (SWfMS) are powerful tools in the automation of scientific experiments. Several workflow executions are necessary to accomplish one scientific experiment. Data provenance, typically collected by SWfMS during workflow execution, is important to understand, reproduce and analyze scientific experiments. Provenance is about data derivation, thus it is typically represented in the form of a directed acyclic graph. For each workflow execution, a provenance gra...

  20. Special Issue: The First Provenance Challenge

    Moreau, Luc; Ludaescher, Bertram T.; Altintas, Ilkay; Barga, Roger S.; Bowers, Shawn; Callahan, Steven P.; Chin, George; Clifford, Ben; Cohen, Shirley; Cohen-Boulakia, Sarah; Davidson, Susan; Deelman, Ewa; digiampietri, Luciano; Foster, Ian T.; Freire, Juliana; Frew, James; Futrelle, Joe; Gibson, Tara D.; Gil, Yolanda; Goble, Carole; Golbeck, Jennifer; Groth, Paul; Holland, David A.; Jiang, Sheng; Kim, Jihie; Koop, David; Krenek, Ales; McPhillips, Timothy; Mehta, Gaurang; Miles, Simon; Metzger, Dominic; Munroe, Steve; Myers, James D.; Plale, Beth A.; Podhorszki, norbert; Ratnakar, Varun; Emanuele , Santos; scheidegger, Carlos E.; Schuchardt, Karen L.; Seltzer, Margo I.; Simmhan, Yogesh L.; Claudio, Silva T.; Slaughter, Peter; Stephan, Eric G.; Stevens, Robert; Turi, Daniele; Vo, Huy T.; Wilde, Mike J.; Zhao, Jun; Zhao, Yong


    The first Provenance Challenge was set up in order to provide a forum for the community to help understand the capabilities of different provenance systems and the expressiveness of their provenance representations. To this end, a Functional Magnetic Resonance Imaging workflow was defined, which participants had to either simulate or run in order to produce some provenance representation, from which a set of identified queries had to be implemented and executed. Sixteen teams responded to the challenge, and submitted their inputs. In this paper, we present the challenge workflow and queries, and summarise the participants contributions.

  1. Src activity increases and Yes activity decreases during mitosis of human colon carcinoma cells.

    Park, J.; Cartwright, C A


    Src and Yes protein-tyrosine kinase activities are elevated in malignant and premalignant tumors of the colon. To determine whether Src activity is elevated throughout the human colon carcinoma cell cycle as it is in polyomavirus middle T antigen- or F527 Src-transformed cells, and whether Yes activity, which is lower than that of Src in the carcinoma cells, is regulated differently, we measured their activities in cycling cells. We observed that the activities of both kinases were higher thr...

  2. SRC-RC结构的push-over分析

    王丹宁; 殷小溦; 王丹



  3. Launch Services, a Proven Model

    Trafton, W. C.; Simpson, J.


    From a commercial perspective, the ability to justify "leap frog" technology such as reusable systems has been difficult to justify because the estimated 5B to 10B investment is not supported in the current flat commercial market coupled with an oversupply of launch service suppliers. The market simply does not justify investment of that magnitude. Currently, next generation Expendable Launch Systems, including Boeing's Delta IV, Lockheed Martin's Atlas 5, Ariane V ESCA and RSC's H-IIA are being introduced into operations signifying that only upgrades to proven systems are planned to meet the changes in anticipated satellite demand (larger satellites, more lifetime, larger volumes, etc.) in the foreseeable future. We do not see a new fleet of ELVs emerging beyond that which is currently being introduced, only continuous upgrades of the fleet to meet the demands. To induce a radical change in the provision of launch services, a Multinational Government investment must be made and justified by World requirements. The commercial market alone cannot justify such an investment. And if an investment is made, we cannot afford to repeat previous mistakes by relying on one system such as shuttle for commercial deployment without having any back-up capability. Other issues that need to be considered are national science and security requirements, which to a large extent fuels the Japanese, Chinese, Indian, Former Soviet Union, European and United States space transportation entries. Additionally, this system must support or replace current Space Transportation Economies with across-the-board benefits. For the next 10 to 20 years, Multinational cooperation will be in the form of piecing together launch components and infrastructure to supplement existing launch systems and reducing the amount of non-recurring investment while meeting the future requirements of the End-User. Virtually all of the current systems have some form of multinational participation: Sea Launch

  4. COOH-terminal association of human smooth muscle calcium channel Ca(v)1.2b with Src kinase protein binding domains: effect of nitrotyrosylation.

    Kang, Minho; Ross, Gracious R; Akbarali, Hamid I


    The carboxyl terminus of the calcium channel plays an important role in the regulation of calcium entry, signal transduction, and gene expression. Potential protein-protein interaction sites within the COOH terminus of the L-type calcium channel include those for the SH3 and SH2 binding domains of c-Src kinase that regulates calcium currents in smooth muscle. In this study, we examined the binding sites involved in Src kinase-mediated phosphorylation of the human voltage-gated calcium channel (Ca(v)) 1.2b (hCav1.2b) and the effect of nitrotyrosylation. Cotransfection of human embryonic kidney (HEK)-293 cells with hCa(v)1.2b and c-Src resulted in tyrosine phosphorylation of the calcium channel, which was prevented by nitration of tyrosine residues by peroxynitrite. Whole cell calcium currents were reduced by 58 + 5% by the Src kinase inhibitor PP2 and 64 + 6% by peroxynitrite. Nitrotyrosylation prevented Src-mediated regulation of the currents. Glutathione S-transferase fusion protein of the distal COOH terminus of hCa(v)1.2b (1809-2138) bound to SH2 domain of Src following tyrosine phosphorylation, while binding to SH3 required the presence of the proline-rich motif. Site-directed mutation of Y(2134) prevented SH2 binding and resulted in reduced phosphorylation of hCa(v)1.2b. Within the distal COOH terminus, single, double, or triple mutations of Y(1837), Y(1861), and Y(2134) were constructed and expressed in HEK-293 cells. The inhibitory effects of PP2 and peroxynitrite on calcium currents were significantly reduced in the double mutant Y(1837-2134F). These data demonstrate that the COOH terminus of hCa(v)1.2b contains sites for the SH2 and SH3 binding of Src kinase. Nitrotyrosylation of these sites prevents Src kinase regulation and may be importantly involved in calcium influx regulation during inflammation.

  5. Active Provenance in Data-intensive Research

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


    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

  6. c-src activating mutation analysis in Chinese patients with colorectal cancer

    Ye-Xiong Tan; Han-Tao Wang; Peng Zhang; Zhong-Hua Yan; Guan-Long Dai; Meng-Chao Wu; Hong-Yang Wang


    AIM: To investigate the occurrence of cellular src (c-src)activating mutation at codon 531 in colorectal cancer patients from Chinese mainland.METHODS: Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay followed by sequencing and single-strand conformation polymorphism analysis were carried out to screen 110 samples of primary colorectal cancer and 20 colorectal liver metastases.RESULTS: Only one sample showed PCR-RFLP-positive results and carried somatic codon 531 mutations. No additional mutation of c-src exon 12 was found.CONCLUSION: c-src codon 531 mutation in colorectal cancer is not the cause of c-src activation.

  7. Generation and Validation of a Mouse Line with a Floxed SRC-3/AIB1 Allele for Conditional Knockout

    Zhaoliang Liu, Lan Liao, Suoling Zhou, Jianming Xu


    Full Text Available The steroid receptor coactivator-3 (SRC-3, also known as AIB1, ACTR, p/CIP and NCOA3, is a transcriptional coactivator for nuclear receptors and certain other transcription factors. SRC-3 is widely expressed and plays important physiological functions and pathogenic roles in breast and prostate cancers. SRC-3 knockout (SRC-3-/- mice display genetic background-dependent embryonic lethality and multiple local and systemic abnormalities. Since both the partial lethality and the systemic effects caused by global SRC-3 knockout interfere with downstream investigation of tissue-specific function of SRC-3, we have generated floxed SRC-3 (SRC-3f/f mice with conditional alleles carrying loxP sites in introns 10 and 12 by a gene-targeting strategy. The two SRC-3f/f mouse lines (A and B are indistinguishable from wild type mice. To test if deletion of the floxed exons 11 and 12 for SRC-3 nuclear receptor interaction domains and disruption of its downstream sequence for transcriptional activation domains would inactivate SRC-3 function, SRC-3f/f mice were crossbred with EIIa-Cre mice to generate SRC-3d/d mice with germ line deletion of the floxed SRC-3 gene. Both lines of SRC-3d/d mice exhibited growth retardation and low IGF-I levels, which was similar to that observed in SRC-3-/- mice. The line A SRC-3d/d mice showed normal viability, while line B SRC-3d/d mice showed partial lethality similar to SRC-3-/- mice, probably due to variable distributions of genetic background during breeding. These results demonstrate that the floxed SRC-3 mouse lines have been successfully established. These mice will be useful for investigating the cell type- and developmental stage-specific functions of SRC-3.

  8. Cytokinesis Failure Leading to Chromosome Instability in v-Src-Induced Oncogenesis.

    Nakayama, Yuji; Soeda, Shuhei; Ikeuchi, Masayoshi; Kakae, Keiko; Yamaguchi, Naoto


    v-Src, an oncogene found in Rous sarcoma virus, is a constitutively active variant of c-Src. Activation of Src is observed frequently in colorectal and breast cancers, and is critical in tumor progression through multiple processes. However, in some experimental conditions, v-Src causes growth suppression and apoptosis. In this review, we highlight recent progress in our understanding of cytokinesis failure and the attenuation of the tetraploidy checkpoint in v-Src-expressing cells. v-Src induces cell cycle changes-such as the accumulation of the 4N cell population-and increases the number of binucleated cells, which is accompanied by an excess number of centrosomes. Time-lapse analysis of v-Src-expressing cells showed that cytokinesis failure is caused by cleavage furrow regression. Microscopic analysis revealed that v-Src induces delocalization of cytokinesis regulators including Aurora B and Mklp1. Tetraploid cell formation is one of the causes of chromosome instability; however, tetraploid cells can be eliminated at the tetraploidy checkpoint. Interestingly, v-Src weakens the tetraploidy checkpoint by inhibiting the nuclear exclusion of the transcription coactivator YAP, which is downstream of the Hippo pathway and its nuclear exclusion is critical in the tetraploidy checkpoint. We also discuss the relationship between v-Src-induced chromosome instability and growth suppression in v-Src-induced oncogenesis.

  9. Archives and societal provenance Australian essays

    Piggott, Michael


    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

  10. Provenance trials of larch in Siberia

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


    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

  11. Provenance Storage, Querying, and Visualization in PBase

    Kianmajd, Parisa [University of California, Davis; Ludascher, Bertram [University of California, Davis; Missier, Paolo [Newcastle University, UK; Chirigati, Fernando [New York University; Wei, Yaxing [ORNL; Koop, David [New York University; Dey, Saumen [University of California, Davis


    We present PBase, a repository for scientific workflows and their corresponding provenance information that facilitates the sharing of experiments among the scientific community. PBase is interoperable since it uses ProvONE, a standard provenance model for scientific workflows. Workflows and traces are stored in RDF, and with the support of SPARQL and the tree cover encoding, the repository provides a scalable infrastructure for querying the provenance data. Furthermore, through its user interface, it is possible to: visualize workflows and execution traces; visualize reachability relations within these traces; issue SPARQL queries; and visualize query results.

  12. Subcellular localization of total and activated Src kinase in African American and Caucasian breast cancer.

    Muralidharan Anbalagan

    Full Text Available BACKGROUND: Src, a non-receptor tyrosine kinase is elevated in cancer with expression and activity correlated with cell proliferation, adhesion, survival, motility, metastasis and angiogenesis. There is limited data on Src expression and subcellular localization in breast cancer and no information about expression in racial/ethnic groups. METHODOLOGY/PRINCIPAL FINDINGS: The present study evaluated Src expression, activity, and subcellular localization in triple negative breast cancer (TNBC and ERα positive breast cancer (ER+BC, cancer tissue and adjacent normal epithelial ducts, and Caucasian and African American cases. 79 paraffin embedded breast carcinoma cases were obtained from Tulane University Hospital between 2007-2009. 39 cases represented TNBC (33-African Americans, 4-Caucasians, 2-unknowns and 40 cases represented ER+BC (21-African Americans, 16-Caucasians, 3-unknowns. Immunohistochemistry was used to measure staining distribution and intensity of total Src and activated phospho-SrcY416 (p-Y416Src in carcinoma tissue and adjacent normal mammary ducts. In TNBC and ER+BC, total Src was significantly higher in cancer compared to adjacent normal ducts (P<0.0001 in both cell membrane and cytoplasm. In membranes, p-Y416Src was elevated in cancer compared to normal ducts. Total Src in the tumor cytoplasm was significantly higher in TNBC compared to ER+BC (P = 0.0028; conversely, p-Y416Src in the tumor cell membranes was higher in TNBC compared to ER+BC (P = 0.0106. Comparison between African American (n = 21 and Caucasian ER+BC (n = 16 revealed no significant difference in expression and localization of total Src and p-Y416Src. TNBC cases positive for lymph node metastasis showed elevated membrane p-Y416Src compared to lymph node negative TNBC (P = 0.027. CONCLUSION/SIGNIFICANCE: Total Src and p-Y416Src were expressed higher in cancer compared to adjacent normal ducts. Cytoplasmic total Src and membrane p-Y416Src were

  13. Causality and the Semantics of Provenance

    James Cheney


    Full Text Available Provenance, or information about the sources, derivation, custody or history of data, has been studied recently in a number of contexts, including databases, scientific workflows and the Semantic Web. Many provenance mechanisms have been developed, motivated by informal notions such as influence, dependence, explanation and causality. However, there has been little study of whether these mechanisms formally satisfy appropriate policies or even how to formalize relevant motivating concepts such as causality. We contend that mathematical models of these concepts are needed to justify and compare provenance techniques. In this paper we review a theory of causality based on structural models that has been developed in artificial intelligence, and describe work in progress on using causality to give a semantics to provenance graphs.

  14. A Provenance Tracking Model for Data Updates

    Gabriel Ciobanu


    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.

  15. Secure Location Provenance for Mobile Devices


    SECURE LOCATION PROVENANCE FOR MOBILE DEVICES UNIVERSITY OF ALABAMA AT BIRMINGHAM JULY 2015 FINAL TECHNICAL REPORT...PROVENANCE FOR MOBILE DEVICES 5a. CONTRACT NUMBER FA8750-12-2-0254 5b. GRANT NUMBER N/A 5c. PROGRAM ELEMENT NUMBER 69220K 6. AUTHOR(S) Ragib Hasan...based services allow mobile device users to access various services based on the users’ current physical location information. Path-critical applications

  16. v-Src Causes Chromosome Bridges in a Caffeine-Sensitive Manner by Generating DNA Damage.

    Ikeuchi, Masayoshi; Fukumoto, Yasunori; Honda, Takuya; Kuga, Takahisa; Saito, Youhei; Yamaguchi, Naoto; Nakayama, Yuji


    An increase in Src activity is commonly observed in epithelial cancers. Aberrant activation of the kinase activity is associated with malignant progression. However, the mechanisms that underlie the Src-induced malignant progression of cancer are not completely understood. We show here that v-Src, an oncogene that was first identified from a Rous sarcoma virus and a mutant variant of c-Src, leads to an increase in the number of anaphase and telophase cells having chromosome bridges. v-Src increases the number of γH2AX foci, and this increase is inhibited by treatment with PP2, a Src kinase inhibitor. v-Src induces the phosphorylation of KAP1 at Ser824, Chk2 at Thr68, and Chk1 at Ser345, suggesting the activation of the ATM/ATR pathway. Caffeine decreases the number of cells having chromosome bridges at a concentration incapable of inhibiting Chk1 phosphorylation at Ser345. These results suggest that v-Src induces chromosome bridges via generation of DNA damage and the subsequent DNA damage response, possibly by homologous recombination. A chromosome bridge gives rise to the accumulation of DNA damage directly through chromosome breakage and indirectly through cytokinesis failure-induced multinucleation. We propose that v-Src-induced chromosome bridge formation is one of the causes of the v-Src-induced malignant progression of cancer cells.

  17. Src promotes cutaneous wound healing by regulating MMP-2 through the ERK pathway.

    Wu, Xue; Yang, Longlong; Zheng, Zhao; Li, Zhenzhen; Shi, Jihong; Li, Yan; Han, Shichao; Gao, Jianxin; Tang, Chaowu; Su, Linlin; Hu, Dahai


    Wound healing is a highly orchestrated, multistep process, and delayed wound healing is a significant symptomatic clinical problem. Keratinocyte migration and re-epithelialization play the most important roles in wound healing, as they determine the rate of wound healing. In our previous study, we found that Src, one of the oldest proto‑oncogenes encoding a membrane-associated, non-receptor protein tyrosine kinase, promotes keratinocyte migration. We therefore hypothesized that Src promotes wound healing through enhanced keratinocyte migration. In order to test this hypothesis, vectors for overexpressing Src and small interfering RNAs (siRNAs) for silencing of Src were used in the present study. We found that the overexpression of Src accelerated keratinocyte migration in vitro and promoted wound healing in vivo without exerting a marked effect on cell proliferation. The extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK) signaling pathways play important roles in Src-accelerated keratinocyte migration. Further experiments demonstrated that Src induced the protein expression of matrix metalloproteinase-2 (MMP-2) and decreased the protein expression of E-cadherin. We suggest that ERK signaling is involved in the Src-mediated regulation of MMP-2 expression. The present study provided evidence that Src promotes keratinocyte migration and cutaneous wound healing, in which the regulation of MMP-2 through the ERK pathway plays an important role, and thus we also demonstrated a potential therapeutic role for Src in cutaneous wound healing.

  18. SRC-3 Has a Role in Cancer Other Than as a Nuclear Receptor Coactivator

    Gang Ma, Yu Ren, Ke Wang, Jianjun He


    Full Text Available Steroid receptor coactivator-3 (SRC-3, also known as AIB1, is a member of the p160 steroid receptor coactivator family. Since SRC-3 was found to be amplified in breast cancer in 1997, the role of SRC-3 in cancer has been broadly investigated. SRC-3 initially was identified as a transcriptional coactivator for nuclear receptors such as the estrogen receptor (ER, involved in the proliferation of hormone-dependent cancers. However, increasing clinical evidence shows that dysregulation of SRC-3 expression in several human hormone-independent cancers is correlated with pathological factors and clinical prognosis. Recently, both in vivo and in vitro studies demonstrate that SRC-3 may influence a number of cancer cellular processes in several ways independent of nuclear receptor signaling. In addition, an SRC-3 transgenic mice model shows that SRC-3 induces tumors in several mouse tissues. These results indicate that the role of SRC-3 in cancer is not just as a nuclear receptor coactivator. The focus of this review is to examine possible SRC-3 roles in cancer, other than as a nuclear receptor coactivator.

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

    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


    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.

  20. Harvester development for new high yielding SRC crops and markets

    Paulson, Mark


    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. Revised SRC-I project baseline. Volume 1


    International Coal Refining Company (ICRC), in cooperation with the Commonwealth of Kentucky has contracted with the United States Department of Energy (DOE) to design, build and operate a first-of-its-kind plant demonstrating the economic, environmental, socioeconomic and technical feasibility of the direct coal liquefaction process known as SRC-I. ICRC has made a massive commitment of time and expertise to design processes, plan and formulate policy, schedules, costs and technical drawings for all plant systems. These fully integrated plans comprise the Project Baseline and are the basis for all future detailed engineering, plant construction, operation, and other work set forth in the contract between ICRC and the DOE. Volumes I and II of the accompanying documents constitute the updated Project Baseline for the SRC-I two-stage liquefaction plant. International Coal Refining Company believes this versatile plant design incorporates the most advanced coal liquefaction system available in the synthetic fuels field. SRC-I two-stage liquefaction, as developed by ICRC, is the way of the future in coal liquefaction because of its product slate flexibility, high process thermal efficiency, and low consumption of hydrogen. The SRC-I Project Baseline design also has made important state-of-the-art advances in areas such as environmental control systems. Because of a lack of funding, the DOE has curtailed the total project effort without specifying a definite renewal date. This precludes the development of revised accurate and meaningful schedules and, hence, escalated project costs. ICRC has revised and updated the original Design Baseline to include in the technical documentation all of the approved but previously non-incorporated Category B and C and new Post-Baseline Engineering Change Proposals.

  2. In Medium Nucleon Structure Functions, SRC, and the EMC effect

    Hen, O; Gilad, S; Wood, S A


    A proposal approved by the Jefferson Lab PAC to study semi-inclusive deep inelastic scattering (DIS) off the deuteron, tagged with high momentum recoiling protons or neutrons emitted at large angle relative to the momentum transfer. This experiment aims at studying the virtuality dependence of the bound nucleon structure function as a possible cause to the EMC effect and the EMC-SRC correlations. The experiment was approved in 2011 for a total run time of 40 days.

  3. An R package for statistical provenance analysis

    Vermeesch, Pieter; Resentini, Alberto; Garzanti, Eduardo


    This paper introduces provenance, a software package within the statistical programming environment R, which aims to facilitate the visualisation and interpretation of large amounts of sedimentary provenance data, including mineralogical, petrographic, chemical and isotopic provenance proxies, or any combination of these. provenance comprises functions to: (a) calculate the sample size required to achieve a given detection limit; (b) plot distributional data such as detrital zircon U-Pb age spectra as Cumulative Age Distributions (CADs) or adaptive Kernel Density Estimates (KDEs); (c) plot compositional data as pie charts or ternary diagrams; (d) correct the effects of hydraulic sorting on sandstone petrography and heavy mineral composition; (e) assess the settling equivalence of detrital minerals and grain-size dependence of sediment composition; (f) quantify the dissimilarity between distributional data using the Kolmogorov-Smirnov and Sircombe-Hazelton distances, or between compositional data using the Aitchison and Bray-Curtis distances; (e) interpret multi-sample datasets by means of (classical and nonmetric) Multidimensional Scaling (MDS) and Principal Component Analysis (PCA); and (f) simplify the interpretation of multi-method datasets by means of Generalised Procrustes Analysis (GPA) and 3-way MDS. All these tools can be accessed through an intuitive query-based user interface, which does not require knowledge of the R programming language. provenance is free software released under the GPL-2 licence and will be further expanded based on user feedback.

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

    Sichun Yang


    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.

  5. Sediment generation and provenance: processes and pathways

    Caracciolo, L.; Garzanti, E.; von Eynatten, H.; Weltje, G. J.


    The ability to trace sediments from their sources to sedimentary basins is a prerequisite for quantitative analysis of Earth-surface dynamics. The comparatively recent revival of sedimentary provenance analysis goes hand-in-hand with the ever expanding range of analytical tools available for quantifying sediment properties (isotopic, mineral, chemical, and petrographic composition, grain-size and shape distributions, age spectra, etc.), and for interpreting such data in paleo-geographic, -tectonic and -climatic terms. The breakdown of sediment budgets into source-specific contributions - one of the most important tasks of provenance analysis - permits quantification of rates of surface processes in the geological past ("deep time"), even in cases where the source areas themselves have been destroyed by global tectonics. Quantitative sedimentary provenance analysis is therefore crucial to the reconstruction of ancient sediment-routing systems, the fundamental units of mass transfer at the Earth's surface.

  6. Src蛋白研究进展%Progress in Src protein

    谢捷; 龚兴国; 曾冬云


    Src is a non - receptor protein tyrosine kinase activated by a number of extracellular signal moleculars. It is recruited to peripheral sites through myristoylation and the SH3 domain. Src initiates intracel-lular signal trandsduction pathways that influence cell adhesion, migration, growth, differentiation and survival though catalytic domain. Src is normally maintained in an inactive conformation because of carboxy terminal Src kinase, but can be activated transiently during cellular events such as mitosis or constitutively by abnormal events such as mutation and some cancers. In additions, c - Sre protein is found to be highly activated and the Src gene is frequently over- expressed in many cancers. These findings suggest that the relationship between c- Src activation/over - expression and cancer progression appears to be significant.

  7. Cullin 3 mediates SRC-3 ubiquitination and degradation to control the retinoic acid response.

    Ferry, Christine; Gaouar, Samia; Fischer, Benoit; Boeglin, Marcel; Paul, Nicodeme; Samarut, Eric; Piskunov, Aleksandr; Pankotai-Bodo, Gabriella; Brino, Laurent; Rochette-Egly, Cecile


    SRC-3 is an important coactivator of nuclear receptors including the retinoic acid (RA) receptor α. Most of SRC-3 functions are facilitated by changes in the posttranslational code of the protein that involves mainly phosphorylation and ubiquitination. We recently reported that SRC-3 is degraded by the proteasome in response to RA. Here, by using an RNAi E3-ubiquitin ligase entry screen, we identified CUL-3 and RBX1 as components of the E3 ubiquitin ligase involved in the RA-induced ubiquitination and subsequent degradation of SRC-3. We also show that the RA-induced ubiquitination of SRC-3 depends on its prior phosphorylation at serine 860 that promotes binding of the CUL-3-based E3 ligase in the nucleus. Finally, phosphorylation, ubiquitination, and degradation of SRC-3 cooperate to control the dynamics of transcription. In all, this process participates to the antiproliferative effect of RA.

  8. Model-based Abstraction of Data Provenance

    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 exte......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...

  9. Identificaiton of Shc Src Homology 2 Domain-Binding Peptoid – Peptide Hybrids

    Choi, Won Jun; Kim, Sung Eun; Stephen, Andrew G.; Weidlich, Iwona; Giubellino, Alessio; Liu, Fa; Worthy, Karen M.; Bindu, Lakshman; Fivash, Matthew J.; Nicklaus, Marc C.; Bottaro, Donald P.; Fisher, Robert J.; Burke, Terrence R.


    A fluorescence anisotropy (FA) competition – based Shc Src homology 2 (SH2) domain-binding was established using the high affinity fluorescein isothiocyanate (FITC)-containing peptide, FITC-NH-(CH2)4-CO-pY-Q-G-L-S-amide (8; Kd = 0.35 μM). Examination of a series of open – chain bis-alkenylamide containing peptides, prepared as ring – closing metathesis precursors, showed that the highest affinities were obtained by replacement of the original Gly residue with Nα-substituted Gly (NSG) “peptoid” residues. This provided peptoid-peptide hybrids of the form, “Ac-pY-Q-[NSG]-L-amide.” Depending on the NSG substituent, certain of these hybrids exhibited up to 40 – fold higher Shc SH2 domain binding affinity than the parent Gly-containing peptide (IC50 = 248 μM), (for example, N-homo-allyl analogue 50; IC50 = 6 μM). To our knowledge, this work represents the first successful example of the application of peptoid-peptide hybrids in the design of SH2 domain-binding antagonists. These results could provide a foundation for further structural optimization of Shc SH2 domain-binding peptide mimetics. PMID:19226165

  10. Identification of Shc Src homology 2 domain-binding peptoid-peptide hybrids.

    Choi, Won Jun; Kim, Sung-Eun; Stephen, Andrew G; Weidlich, Iwona; Giubellino, Alessio; Liu, Fa; Worthy, Karen M; Bindu, Lakshman; Fivash, Matthew J; Nicklaus, Marc C; Bottaro, Donald P; Fisher, Robert J; Burke, Terrence R


    A fluorescence anisotropy (FA) competition-based Shc Src homology 2 (SH2) domain-binding was established using the high affinity fluorescein isothiocyanate (FITC) containing peptide, FITC-NH-(CH2)4-CO-pY-Q-G-L-S-amide (8; Kd = 0.35 microM). Examination of a series of open-chain bis-alkenylamide containing peptides, prepared as ring-closing metathesis precursors, showed that the highest affinities were obtained by replacement of the original Gly residue with N alpha-substituted Gly (NSG) "peptoid" residues. This provided peptoid-peptide hybrids of the form "Ac-pY-Q-[NSG]-L-amide." Depending on the NSG substituent, certain of these hybrids exhibited up to 40-fold higher Shc SH2 domain-binding affinity than the parent Gly-containing peptide (IC50 = 248 microM) (for example, for N-homoallyl analogue 50, IC50 = 6 microM). To our knowledge, this work represents the first successful example of the application of peptoid-peptide hybrids in the design of SH2 domain-binding antagonists. These results could provide a foundation for further structural optimization of Shc SH2 domain-binding peptide mimetics.

  11. Crk adaptor protein-induced phosphorylation of Gab1 on tyrosine 307 via Src is important for organization of focal adhesions and enhanced cell migration

    Takuya Watanabe; Masumi Tsuda; Yoshinori Makino; Tassos Konstantinou; Hiroshi Nishihara; Tokifumi Majima; Akio Minami; Stephan M Feller; Shinya Tanaka


    Upon growth factor stimulation, the scaffold protein, Gabl, is tyrosine phosphorylated and subsequently the adaptor protein, Crk, transmits signals from Gabl. We have previously shown that Crk overexpression, which is detectable in various human cancers, induces tyrosine phosphorylation of Gab1 without extraceilular stimuli. In the present study, the underlying mechanisms were further investigated. Mutational analyses of Crkll demonstrated that the SH2 domain, but not the SH3(N) or the regulatory Y221 residue of Crkll, is critical for the induction of Gabl-Y307 phosphorylation. SH2 mutation of Crkll also decreased the interaction with Gab1. In GST pull-down assay, Crk-SH2 bound to wild-type Gabl, whereas Crk-SH3(N) interacted with the Gabl mutant, which lacks the clus-tered tyrosine region (residues 242-410). Tyrosine phosphorylation of Gabl was induced by all Crk family proteins, but not other SH2-containing signalling adaptors. Src-family kinase inhibitor, PP2, abrogates Crk-induced tyrosine phosphorylations of Gabl. Y307 phosphorylation was undetectable in fibroblasts lacking Src, Yes, and Fyn, even upon overexpression of Crk, whereas cells lacking only Yes and Fyn still contained Gabl with phosphorylated Y307. Furthermore, Crk induced the phosphorylation of Src-Y416; accordingly the interaction between Crk and Csk was increased. The GabI-Y307F mutant failed to localize near the plasma membrane even upon HGF stimulation and decreased cell migration. Moreover, Gabl-Y307F disturbed the localization of Crk, FAK, and paxiilin, which are the typical components of focal adhesions. Taken together, these results indicate that Crk facilitates tyrosine phosphory-lation of Gabl-Y307 through Src, contributing to the organization of focal adhesions and enhanced cell migration, thereby possibly promoting human cancer development.

  12. The Role of c-Src Activation in Prostate Tumor Progression


    expressing a neomycin - resistance gene) that we have used in the past in other cell types (Windharn et al., 2002). G418- resistant colonies were isolated...ectopically expressed chicken Src; with an antibody specific fordirectly leads to increased VEGF phosphorylated tyr 416 to estimate Src activation, and with...antibody, specific for chicken c-Src. We first designed and selected clones with the activating Y527F mutation, as they would be expected to be more

  13. Berberine reduces Toll-like receptor-mediated macrophage migration by suppression of Src enhancement.

    Cheng, Wei-Erh; Ying Chang, Miao; Wei, Jyun-Yan; Chen, Yen-Jen; Maa, Ming-Chei; Leu, Tzeng-Horng


    Berberine is an isoquinoline with anti-inflammatory activity. We previously demonstrated that there was a loop of signal amplification between nuclear factor kappa B and Src for macrophage mobility triggered by the engagement of Toll-like receptors (TLRs). The simultaneous suppression of lipopolysaccharide (LPS)-mediated upregulation of inducible nitric oxide synthase, cyclooxygenase 2, and cell mobility in berberine-treated macrophages suggested Src might be a target of berberine. Indeed, th reduced migration, greatly suppressed Src induction in both protein and RNA transcript by berberine were observed in macrophages exposed to LPS, peptidoglycan, polyinosinic-polycytidylic acid, and CpG-oligodeoxynucleotides. In addition to Src induction, berberine also inhibited LPS-mediated Src activation in Src overexpressing macrophages and S-nitroso-N-acetylpenicillamine (a nitric oxide donor) could partly restore it. Moreover, berberine suppressed Src activity in fibronectin-stimulated macrophages and in v-Src transformed cells. These results implied that by effectively reducing Src expression and activity, berberine inhibited TLR-mediated cell motility in macrophages.

  14. Coactivator SRC-2-dependent metabolic reprogramming mediates prostate cancer survival and metastasis.

    Dasgupta, Subhamoy; Putluri, Nagireddy; Long, Weiwen; Zhang, Bin; Wang, Jianghua; Kaushik, Akash K; Arnold, James M; Bhowmik, Salil K; Stashi, Erin; Brennan, Christine A; Rajapakshe, Kimal; Coarfa, Cristian; Mitsiades, Nicholas; Ittmann, Michael M; Chinnaiyan, Arul M; Sreekumar, Arun; O'Malley, Bert W


    Metabolic pathway reprogramming is a hallmark of cancer cell growth and survival and supports the anabolic and energetic demands of these rapidly dividing cells. The underlying regulators of the tumor metabolic program are not completely understood; however, these factors have potential as cancer therapy targets. Here, we determined that upregulation of the oncogenic transcriptional coregulator steroid receptor coactivator 2 (SRC-2), also known as NCOA2, drives glutamine-dependent de novo lipogenesis, which supports tumor cell survival and eventual metastasis. SRC-2 was highly elevated in a variety of tumors, especially in prostate cancer, in which SRC-2 was amplified and overexpressed in 37% of the metastatic tumors evaluated. In prostate cancer cells, SRC-2 stimulated reductive carboxylation of α-ketoglutarate to generate citrate via retrograde TCA cycling, promoting lipogenesis and reprogramming of glutamine metabolism. Glutamine-mediated nutrient signaling activated SRC-2 via mTORC1-dependent phosphorylation, which then triggered downstream transcriptional responses by coactivating SREBP-1, which subsequently enhanced lipogenic enzyme expression. Metabolic profiling of human prostate tumors identified a massive increase in the SRC-2-driven metabolic signature in metastatic tumors compared with that seen in localized tumors, further implicating SRC-2 as a prominent metabolic coordinator of cancer metastasis. Moreover, SRC-2 inhibition in murine models severely attenuated the survival, growth, and metastasis of prostate cancer. Together, these results suggest that the SRC-2 pathway has potential as a therapeutic target for prostate cancer.

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

    Lee Jang


    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.

  16. Elastic-plastic study on high building with SRC transferring story


    A new type of transferring structure for steel reinforced concrete (SRC) beams is used in high building. The pushover analysis method was used to study the failure mechanism and ductility of SRC transferring structure through consulting pseudo-static test results for the structure. And, the occurrence order and position of the plastic hinge, the weak story and seismic capacity of high building with SRC transferring story were also studied through consulting shaking table test results for the high building, showing that the seismic behavior of high building with SRC transferring story is good.

  17. geographic characterisation of african provenances of

    Polymerase Chain Reaction (PCR) analysis based on Internal Franscribed Spacers ... d'origine de l'espèce et (ii) déterminer l'ampleur de la diversité génétique au sein de F. ... of basic knowledge on the amount of genetic ... variation in genetic structure within provenances ... DNA especiallytheribosomal RNA(rRNA) genes.

  18. The PBase Scientific Workflow Provenance Repository

    Víctor Cuevas-Vicenttín


    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.

  19. Provenance Context Entity (PaCE): Scalable Provenance Tracking for Scientific RDF Data.

    Sahoo, Satya S; Bodenreider, Olivier; Hitzler, Pascal; Sheth, Amit; Thirunarayan, Krishnaprasad

    The Resource Description Framework (RDF) format is being used by a large number of scientific applications to store and disseminate their datasets. The provenance information, describing the source or lineage of the datasets, is playing an increasingly significant role in ensuring data quality, computing trust value of the datasets, and ranking query results. Current provenance tracking approaches using the RDF reification vocabulary suffer from a number of known issues, including lack of formal semantics, use of blank nodes, and application-dependent interpretation of reified RDF triples. In this paper, we introduce a new approach called Provenance Context Entity (PaCE) that uses the notion of provenance context to create provenance-aware RDF triples. We also define the formal semantics of PaCE through a simple extension of the existing RDF(S) semantics that ensures compatibility of PaCE with existing Semantic Web tools and implementations. We have implemented the PaCE approach in the Biomedical Knowledge Repository (BKR) project at the US National Library of Medicine. The evaluations demonstrate a minimum of 49% reduction in total number of provenance-specific RDF triples generated using the PaCE approach as compared to RDF reification. In addition, performance for complex queries improves by three orders of magnitude and remains comparable to the RDF reification approach for simpler provenance queries.

  20. Titian: Data Provenance Support in Spark.

    Interlandi, Matteo; Shah, Kshitij; Tetali, Sai Deep; Gulzar, Muhammad Ali; Yoo, Seunghyun; Kim, Miryung; Millstein, Todd; Condie, Tyson


    Debugging data processing logic in Data-Intensive Scalable Computing (DISC) systems is a difficult and time consuming effort. Today's DISC systems offer very little tooling for debugging programs, and as a result programmers spend countless hours collecting evidence (e.g., from log files) and performing trial and error debugging. To aid this effort, we built Titian, a library that enables data provenance-tracking data through transformations-in Apache Spark. Data scientists using the Titian Spark extension will be able to quickly identify the input data at the root cause of a potential bug or outlier result. Titian is built directly into the Spark platform and offers data provenance support at interactive speeds-orders-of-magnitude faster than alternative solutions-while minimally impacting Spark job performance; observed overheads for capturing data lineage rarely exceed 30% above the baseline job execution time.

  1. Obsidian provenance research in the Americas.

    Glascock, Michael D


    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.

  2. Leveraging Data Provenance to Enhance Cyber Resilience


    1 Leveraging Data Provenance to Enhance Cyber Resilience Thomas Moyer∗, Patrick Cable∗, Karishma Chadha∗, Robert Cunningham∗, Nabil Schear∗, Warren...emphasize this. It is untenable to assume that a system even with designed-in security can successfully repel all attacks. The next generation of secure must also be able to withstand successful attacks using cyber resilience. Cyber re- silience broadly encompasses many areas including traditional

  3. Src mutation induces acquired lapatinib resistance in ERBB2-amplified human gastroesophageal adenocarcinoma models.

    Yong Sang Hong

    Full Text Available ERBB2-directed therapy is now a routine component of therapy for ERBB2-amplified metastatic gastroesophageal adenocarcinomas. However, there is little knowledge of the mechanisms by which these tumors develop acquired resistance to ERBB2 inhibition. To investigate this question we sought to characterize cell line models of ERBB2-amplified gastroesophageal adenocarcinoma with acquired resistance to ERBB2 inhibition. We generated lapatinib-resistant (LR subclones from an initially lapatinib-sensitive ERBB2-amplified esophageal adenocarcinoma cell line, OE19. We subsequently performed genomic characterization and functional analyses of resistant subclones with acquired lapatinib resistance. We identified a novel, acquired SrcE527K mutation in a subset of LR OE19 subclones. Cells with this mutant allele harbour increased Src phosphorylation. Genetic and pharmacologic inhibition of Src resensitized these subclones to lapatinib. Biochemically, Src mutations could activate both the phosphatidylinositol 3-kinase and mitogen activated protein kinase pathways in the lapatinib-treated LR OE19 cells. Ectopic expression of SrcE527K mutation also was sufficient to induce lapatinib resistance in drug-naïve cells. These results indicate that pathologic activation of Src is a potential mechanism of acquired resistance to ERBB2 inhibition in ERBB2-amplified gastroesophageal cancer. Although Src mutation has not been described in primary tumor samples, we propose that the Src hyperactivation should be investigated in the settings of acquired resistance to ERBB2 inhibition in esophageal and gastric adenocarcinoma.

  4. 76 FR 21404 - National Park Service Alaska Region's Subsistence Resource Commission (SRC) Program


    ... National Park Service National Park Service Alaska Region's Subsistence Resource Commission (SRC) Program AGENCY: National Park Service, Interior. ACTION: Notice of public meeting for the National Park Service... Park SRC will meet to develop and continue work on National Park Service (NPS) subsistence...

  5. Inhibition of epithelial to mesenchymal transition in metastatic breast carcinoma cells by c-Src suppression.

    Liu, Xiang; Feng, Renqing


    The aberrant activation of c-Src regulates multiple functions during tumor progression. This study was conducted to investigate the role of c-Src suppression in epithelial to mesenchymal transition (EMT) process in human breast carcinoma cells. c-Src suppression by PP2 (a Src-family kinase inhibitor) or small interfering RNA (siRNA) was carried out in MCF-7 and MDA-MB-231 cells. Cell migration was analyzed by wound-healing assay. The transcription and protein levels of EMT markers and transcription factors were evaluated by reverse transcription-PCR and Western blot analysis, respectively. The changed cell morphology was photographed by light microscope. c-Src suppression by PP2 or siRNA reversed the mesenchymal-like phenotype in MDA-MB-231 cells. E-cadherin was upregulated in MCF-7 and MDA-MB-231 cells after c-Src suppression, whereas vimentin was downregulated in MDA-MB-231 cells. Slug and SIP1 were downregulated after c-Src suppression in MCF-7 and MDA-MB-231 cells, whereas Twist was unchanged. These results suggest that c-Src suppression by PP2 or siRNA may inhibit EMT through regulation of different transcription factors in breast carcinoma cells that have different metastatic potential.

  6. SRC-1 quarterly technical report, April-June 1981. [Review of analytical methods needed in SRC Demonstration plants


    Twenty-three papers involving the design, materials and equipment for the SRC-1 demonstration coal liquefaction plant near Newman, Daviess County, Kentucky, have been entered individually into EDB and ERA. A number of the papers deal also with the analytical methodology required for the plant, including a rather detailed evaluation of the accuracy requirements and careful evaluation of several methods such as gas chromatography, mass spectroscopy, nuclear magnetic resonance, etc. Flexibility of design is stressed so that products can be optimized for the market and charged if the market requires different products. (LTN)

  7. Src regulates membrane trafficking of the Kv3.1b channel.

    Bae, Seong Han; Kim, Dong Hyun; Shin, Seok Kyo; Choi, Jin Sung; Park, Kang-Sik


    The Kv3.1 channel plays a crucial role in regulating the high-frequency firing properties of neurons. Here, we determined whether Src regulates the subcellular distributions of the Kv3.1b channel. Co-expression of active Src induced a dramatic redistribution of Kv3.1b to the endoplasmic reticulum. Furthermore, co-expression of the Kv3.1b channel with active Src induced a remarkable decrease in the pool of Kv3.1b at the cell surface. Moreover, the co-expression of active Src results in a significant decrease in the peak current densities of the Kv3.1b channel, and a substantial alteration in the voltage dependence of its steady-state inactivation. Taken together, these results indicate that Src kinase may play an important role in regulating membrane trafficking of Kv3.1b channels.

  8. Seismic cyclic loading test of SRC columns confined with 5-spirals


    Presented herein is an experimental study on seismic resistance of rectangular steel reinforced concrete (SRC) columns confined with a new type of multi-spiral cage. The multi-spiral cage is a device of five interconnected spirals, named "5-spirals", with a large spiral at the center and four small ones at the corners. The innovation of applying the 5-spirals to SRC column is to take its superiority in concrete confinement and efficiency in automatic production for the precast construction industry. Four full-scale SRC columns were tested under horizontal cyclic loading. All of the tested columns were capable of sustaining a drift angle up to 6% radians. The hysteresis loops observed from the cyclic loading tests indicated that the spirally confined SRC columns demonstrated excellent performances in both strength and ductility. The test results suggested that with significant saving of the confinement steel, the newly innovated 5-spirals can be successfully applied to the precast rectangular SRC columns.

  9. Src and PI3 K inhibitors affect the virulence factors of Entamoeba histolytica.

    López-Contreras, L; Hernández-Ramírez, V I; Flores-García, Y; Chávez-Munguía, B; Talamás-Rohana, P


    Protein kinases (PKs) of parasitic protozoa are being evaluated as drug targets. A large number of protein kinases within the protein kinome of Entamoeba histolytica strongly suggest that protein phosphorylation is a key component of pathogenesis regulation by this parasite. PI3 K and Src are kinases previously described in this parasite, but their role is poorly understood. Here, the effect of Src-1-inhibitor and PI3 K inhibitor (Wortmannin) on the virulence factors of E. histolytica was evaluated. Results show that both inhibitors affect the actin cytoskeleton and the amoebic movement. Also, the proteolytic activity is diminished by Wortmannin, but not by Src-inhibitor-1; however, the phagocytic capacity is diminished by Wortmannin and Src-1-inhibitor. Finally, we found that the virulence in vivo of E. histolytica is affected by Wortmannin but not by Src-1-inhibitor. This study opens the way for the design of anti-amoebic drugs based on kinase inhibition.

  10. The afatinib resistance of in vivo generated H1975 lung cancer cell clones is mediated by SRC/ERBB3/c-KIT/c-MET compensatory survival signaling.

    Booth, Laurence; Roberts, Jane L; Tavallai, Mehrad; Webb, Timothy; Leon, Daniel; Chen, Jesse; McGuire, William P; Poklepovic, Andrew; Dent, Paul


    We generated afatinib resistant clones of H1975 lung cancer cells by transient exposure of established tumors to the drug and collected the re-grown tumors. Afatinib resistant H1975 clones did not exhibit any additional mutations in proto-oncogenes when compared to control clones. Afatinib resistant H1975 tumor clones expressed less PTEN than control clones and in afatinib resistant clones this correlated with increased basal SRC Y416, ERBB3 Y1289, AKT T308 and mTOR S2448 phosphorylation, decreased expression of ERBB1, ERBB2 and ERBB3 and increased total expression of c-MET, c-KIT and PDGFRβ. Afatinib resistant clones were selectively killed by knock down of [ERBB3 + c-MET + c-KIT] but not by the individual or doublet knock down combinations. The combination of the ERBB1/2/4 inhibitor afatinib with the SRC family inhibitor dasatinib killed afatinib resistant H1975 cells in a greater than additive fashion; other drugs used in combination with dasatinib such as sunitinib, crizotinib and amufatinib were less effective. [Afatinib + dasatinib] treatment profoundly inactivated ERBB3, AKT and mTOR in the H1975 afatinib resistant clones and increased ATG13 S318 phosphorylation. Knock down of ATG13, Beclin1 or eIF2α strong suppressed killing by [ERBB3 + c-MET + c-KIT] knock down, but were only modestly protective against [afatinib + dasatinib] lethality. Thus afatinib resistant H1975 NSCLC cells rely on ERBB1- and SRC-dependent hyper-activation of residual ERBB3 and elevated signaling, due to elevated protein expression, from wild type c-MET and c-KIT to remain alive. Inhibition of ERBB3 signaling via both blockade of SRC and ERBB1 results in tumor cell death.

  11. Redox-sensitive Akt and Src regulate coronary collateral growth in metabolic syndrome.

    Reed, Ryan; Potter, Barry; Smith, Erika; Jadhav, Rashmi; Villalta, Patricia; Jo, Hanjoong; Rocic, Petra


    We have recently shown that the inability of repetitive ischemia (RI) to activate p38 MAPK (p38) and Akt in metabolic syndrome [JCR:LA-cp (JCR)] rats was associated with impaired coronary collateral growth (CCG). Furthermore, Akt and p38 activation correlated with optimal O(2)(-). levels and were altered in JCR rats, and redox-sensitive p38 activation was required for CCG. Here, we determined whether the activation of Src, a possible upstream regulator, was altered in JCR rats and whether redox-dependent Src and Akt activation were required for CCG. CCG was assessed by myocardial blood flow (microspheres) and kinase activation was assessed by Western blot analysis in the normal zone and collateral-dependent zone (CZ). RI induced Src activation (approximately 3-fold) in healthy [Wistar-Kyoto (WKY)] animals but not in JCR animals. Akt inhibition decreased (approximately 50%), and Src inhibition blocked RI-induced CCG in WKY rats. Src inhibition decreased p38 and Akt activation. Myocardial oxidative stress (O(2)(-). and oxidized/reduced thiols) was measured quantitatively (X-band electron paramagnetic resonance). An antioxidant, apocynin, reduced RI-induced oxidative stress in JCR rats to levels induced by RI in WKY rats versus the reduction in WKY rats to very low levels. This resulted in a significant restoration of p38 (approximately 80%), Akt (approximately 65%), and Src (approximately 90%) activation in JCR rats but decreased the activation in WKY rats (p38: approximately 45%, Akt: approximately 65%, and Src: approximately 100%), correlating with reduced CZ flow in WKY rats (approximately 70%), but significantly restored CZ flow in JCR rats (approximately 75%). We conclude that 1) Akt and Src are required for CCG, 2) Src is a redox-sensitive upstream regulator of RI-induced p38 and Akt activation, and 3) optimal oxidative stress levels are required for RI-induced p38, Akt, and Src activation and CCG.

  12. Regulation of N-Formyl Peptide Receptor Signaling and Trafficking by Arrestin-Src Kinase Interaction.

    Wagener, Brant M; Marjon, Nicole A; Prossnitz, Eric R


    Arrestins were originally described as proteins recruited to ligand-activated, phosphorylated G protein-coupled receptors (GPCRs) to attenuate G protein-mediated signaling. It was later revealed that arrestins also mediate GPCR internalization and recruit a number of signaling proteins including, but not limited to, Src family kinases, ERK1/2, and JNK3. GPCR-arrestin binding and trafficking control the spatial and temporal activity of these multi-protein complexes. In previous reports, we concluded that N-formyl peptide receptor (FPR)-mediated apoptosis, which occurs upon receptor stimulation in the absence of arrestins, is associated with FPR accumulation in perinuclear recycling endosomes. Under these conditions, inhibition of Src kinase and ERK1/2 prevented FPR-mediated apoptosis. To better understand the role of Src kinase in this process, in the current study we employed a previously described arrestin-2 (arr2) mutant deficient in Src kinase binding (arr2-P91G/P121E). Unlike wild type arrestin, arr2-P91G/P121E did not inhibit FPR-mediated apoptosis, suggesting that Src binding to arrestin-2 prevents apoptotic signaling. However, in cells expressing this mutant, FPR-mediated apoptosis was still blocked by inhibition of Src kinase activity, suggesting that activation of Src independent of arrestin-2 binding is involved in FPR-mediated apoptosis. Finally, while Src kinase inhibition prevented FPR-mediated-apoptosis in the presence of arr2-P91G/P121E, it did not prevent FPR-arr2-P91G/P121E accumulation in the perinuclear recycling endosome. On the contrary, inhibition of Src kinase activity mediated the accumulation of activated FPR-wild type arrestin-2 in recycling endosomes without initiating FPR-mediated apoptosis. Based on these observations, we conclude that Src kinase has two independent roles following FPR activation that regulate both FPR-arrestin-2 signaling and trafficking.

  13. Regulation of N-Formyl Peptide Receptor Signaling and Trafficking by Arrestin-Src Kinase Interaction.

    Brant M Wagener

    Full Text Available Arrestins were originally described as proteins recruited to ligand-activated, phosphorylated G protein-coupled receptors (GPCRs to attenuate G protein-mediated signaling. It was later revealed that arrestins also mediate GPCR internalization and recruit a number of signaling proteins including, but not limited to, Src family kinases, ERK1/2, and JNK3. GPCR-arrestin binding and trafficking control the spatial and temporal activity of these multi-protein complexes. In previous reports, we concluded that N-formyl peptide receptor (FPR-mediated apoptosis, which occurs upon receptor stimulation in the absence of arrestins, is associated with FPR accumulation in perinuclear recycling endosomes. Under these conditions, inhibition of Src kinase and ERK1/2 prevented FPR-mediated apoptosis. To better understand the role of Src kinase in this process, in the current study we employed a previously described arrestin-2 (arr2 mutant deficient in Src kinase binding (arr2-P91G/P121E. Unlike wild type arrestin, arr2-P91G/P121E did not inhibit FPR-mediated apoptosis, suggesting that Src binding to arrestin-2 prevents apoptotic signaling. However, in cells expressing this mutant, FPR-mediated apoptosis was still blocked by inhibition of Src kinase activity, suggesting that activation of Src independent of arrestin-2 binding is involved in FPR-mediated apoptosis. Finally, while Src kinase inhibition prevented FPR-mediated-apoptosis in the presence of arr2-P91G/P121E, it did not prevent FPR-arr2-P91G/P121E accumulation in the perinuclear recycling endosome. On the contrary, inhibition of Src kinase activity mediated the accumulation of activated FPR-wild type arrestin-2 in recycling endosomes without initiating FPR-mediated apoptosis. Based on these observations, we conclude that Src kinase has two independent roles following FPR activation that regulate both FPR-arrestin-2 signaling and trafficking.

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

    Jan eGörtzen


    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

  15. Residuation theory

    Blyth, T S; Sneddon, I N; Stark, M


    Residuation Theory aims to contribute to literature in the field of ordered algebraic structures, especially on the subject of residual mappings. The book is divided into three chapters. Chapter 1 focuses on ordered sets; directed sets; semilattices; lattices; and complete lattices. Chapter 2 tackles Baer rings; Baer semigroups; Foulis semigroups; residual mappings; the notion of involution; and Boolean algebras. Chapter 3 covers residuated groupoids and semigroups; group homomorphic and isotone homomorphic Boolean images of ordered semigroups; Dubreil-Jacotin and Brouwer semigroups; and loli

  16. Data Provenance in Photogrammetry Through Documentation Protocols

    Carboni, N.; Bruseker, G.; Guillem, A.; Bellido Castañeda, D.; Coughenour, C.; Domajnko, M.; de Kramer, M.; Ramos Calles, M. M.; Stathopoulou, E. K.; Suma, R.


    Documenting the relevant aspects in digitisation processes such as photogrammetry in order to provide a robust provenance for their products continues to present a challenge. The creation of a product that can be re-used scientifically requires a framework for consistent, standardised documentation of the entire digitisation pipeline. This article provides an analysis of the problems inherent to such goals and presents a series of protocols to document the various steps of a photogrammetric workflow. We propose this pipeline, with descriptors to track all phases of digital product creation in order to assure data provenance and enable the validation of the operations from an analytic and production perspective. The approach aims to support adopters of the workflow to define procedures with a long term perspective. The conceptual schema we present is founded on an analysis of information and actor exchanges in the digitisation process. The metadata were defined through the synthesis of previous proposals in this area and were tested on a case study. We performed the digitisation of a set of cultural heritage artefacts from an Iron Age burial in Ilmendorf, Germany. The objects were captured and processed using different techniques, including a comparison of different imaging tools and algorithms. This augmented the complexity of the process allowing us to test the flexibility of the schema for documenting complex scenarios. Although we have only presented a photogrammetry digitisation scenario, we claim that our schema is easily applicable to a multitude of 3D documentation processes.

  17. Provenance of sediments from Sumatra, Indonesia

    Liebermann, Christof; Hall, Robert; Gough, Amy


    The island of Sumatra is situated at the south-western margin of the Indonesian archipelago. Sumatra is affected by active continental margin volcanism along the Sunda Trench, west of Sumatra as a result of active northeast subduction of the Indian plate under the Eurasian plate. Exposures of the Palaeozoic meta-sedimentary basement are mainly limited in extent to the northeast-southwest trending Barisan Mountain chain. The younger Cenozoic rocks are widespread across Sumatra, but can be grouped into structurally subdivided 'fore-arc', 'intramontane', and 'back-arc' basins. However, the formation of the basins pre-dates the current magmatic arc, thus a classical arc-related generation model can not be applied. The Cenozoic formations are well studied due to hydrocarbon enrichment, but little is known about their provenance history. A comprehensive sedimentary provenance study of the Cenozoic formations can aid in the wider understanding of Sumatran petroleum plays, can contribute to palaeographic reconstruction of western SE Asia, and might help to simplify the overall stratigraphy of Sumatra. This work represents a multi-proxy provenance study of sedimentary rocks from the main Cenozoic basins of Sumatra, alongside sediment from present-day river systems. The project refines the provenance in two ways: first, by studying the heavy mineral assemblages of the targeted formations, and secondly, by U-Pb detrital zircon dating using LA-ICP-MS to identify the age-range of the potential sediment sources. Preliminary U-Pb zircon age-data of >1500 concordant grains (10% discordant cut-off), heavy mineral compositions, and thin section analysis from two fieldwork seasons indicate a mixed provenance model, with a proximal igneous source, and mature basement rocks. An increase of the proximal signature in Lower-Miocene strata indicated by the occurrence of unstable heavy mineral phases such as apatite, and clinopyroxene suggests a major change of the source at the Oligocene

  18. The coactivator SRC-1 is an essential coordinator of hepatic glucose production.

    Louet, Jean-Francois; Chopra, Atul R; Sagen, Jorn V; An, Jie; York, Brian; Tannour-Louet, Mounia; Saha, Pradip K; Stevens, Robert D; Wenner, Brett R; Ilkayeva, Olga R; Bain, James R; Zhou, Suoling; DeMayo, Franco; Xu, Jianming; Newgard, Christopher B; O'Malley, Bert W


    Gluconeogenesis makes a major contribution to hepatic glucose production, a process critical for survival in mammals. In this study, we identify the p160 family member, SRC-1, as a key coordinator of the hepatic gluconeogenic program in vivo. SRC-1-null mice displayed hypoglycemia secondary to a deficit in hepatic glucose production. Selective re-expression of SRC-1 in the liver restored blood glucose levels to a normal range. SRC-1 was found induced upon fasting to coordinate in a cell-autonomous manner, the gene expression of rate-limiting enzymes of the gluconeogenic pathway. At the molecular level, the main role of SRC-1 was to modulate the expression and the activity of C/EBPα through a feed-forward loop in which SRC-1 used C/EBPα to transactivate pyruvate carboxylase, a crucial gene for initiation of the gluconeogenic program. We propose that SRC-1 acts as a critical mediator of glucose homeostasis in the liver by adjusting the transcriptional activity of key genes involved in the hepatic glucose production machinery. Copyright © 2010 Elsevier Inc. All rights reserved.

  19. Androgen receptor non-nuclear regulation of prostate cancer cell invasion mediated by Src and matriptase.

    Zarif, Jelani C; Lamb, Laura E; Schulz, Veronique V; Nollet, Eric A; Miranti, Cindy K


    Castration-resistant prostate cancers still depend on nuclear androgen receptor (AR) function despite their lack of dependence on exogenous androgen. Second generation anti-androgen therapies are more efficient at blocking nuclear AR; however resistant tumors still develop. Recent studies indicate Src is highly active in these resistant tumors. By manipulating AR activity in several different prostate cancer cell lines through RNAi, drug treatment, and the use of a nuclear-deficient AR mutant, we demonstrate that androgen acting on cytoplasmic AR rapidly stimulates Src tyrosine kinase via a non-genomic mechanism. Cytoplasmic AR, acting through Src enhances laminin integrin-dependent invasion. Active Matriptase, which cleaves laminin, is elevated within minutes after androgen stimulation, and is subsequently shed into the medium. Matriptase activation and shedding induced by cytoplasmic AR is dependent on Src. Concomitantly, CDCP1/gp140, a Matriptase and Src substrate that controls integrin-based migration, is activated. However, only inhibition of Matriptase, but not CDCP1, suppresses the AR/Src-dependent increase in invasion. Matriptase, present in conditioned medium from AR-stimulated cells, is sufficient to enhance invasion in the absence of androgen. Thus, invasion is stimulated by a rapid but sustained increase in Src activity, mediated non-genomically by cytoplasmic AR, leading to rapid activation and shedding of the laminin protease Matriptase.

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

    Zou, Zhengzhi, E-mail: [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: [Xiamen Cancer Center, Department of Medical Oncology, The First Affiliated Hospital of Xiamen University, Xiamen 361000 (China)


    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.

  1. Clinical implications of the coexpression of SRC1 and NANOG in HER-2-overexpressing breast cancers

    Jin CY


    Full Text Available Chengyan Jin,1 Xingyi Zhang,1 Mei Sun,2 Yifan Zhang,1 Guangxin Zhang,1 Bin Wang1 1Department of Thoracic Surgery, 2Department of Pathology, The Second Affiliated Hospital of Jilin University, Changchun, People’s Republic of China Objective: Given the lack of clarity on the expression status of SRC1 protein in breast cancer, we attempted to ascertain the clinical implications of the expression of this protein in breast cancer.Methods: Samples from 312 breast cancer patients who were followed up for 5 years were analyzed in this study. The associations of SRC1 expression and clinicopathological factors with the prognosis of breast cancer were determined.Results: The 312 breast cancer patients underwent radical resection, and 155 (49.68% of them demonstrated high expression of SRC1 protein. No significant differences were found for tumor size, estrogen receptor expression, or progesterone receptor expression (P=0.191, 0.888, or 0.163, respectively. It is noteworthy that SRC1 expression was found to be related to HER-2 and Ki-67 expression (P=0.044 and P=0.001, respectively. According to logistic regression analysis, SRC1 expression was also significantly correlated with Ki-67 and HER-2 expression (P=0.032 and P=0.001, respectively. Survival analysis showed that patients with a high expression of SRC1 and NANOG and those with SRC1 and NANOG coexpression had significantly poorer postoperative disease-specific survival than those with no expression in the HER-2-positive group (P=0.032, 0.01, and P=0.01, respectively.Conclusion: High SRC1 protein expression was related to the prognosis of HER-2-overexpressing breast cancers. Keywords: breast cancer, prognosis, molecular classification, SRC1, NANOG

  2. Drosophila actin-Capping Protein limits JNK activation by the Src proto-oncogene.

    Fernández, B G; Jezowska, B; Janody, F


    The Src family kinases c-Src, and its downstream effectors, the Rho family of small GTPases RhoA and Jun N-terminal kinase (JNK) have a significant role in tumorigenesis. In this report, using the Drosophila wing disc epithelium as a model system, we demonstrate that the actin-Capping Protein (CP) αβ heterodimer, which regulates actin filament (F-actin) polymerization, limits Src-induced apoptosis or tissue overgrowth by restricting JNK activation. We show that overexpressing Src64B drives JNK-independent loss of epithelial integrity and JNK-dependent apoptosis via Btk29A, p120ctn and Rho1. However, when cells are kept alive with the Caspase inhibitor P35, JNK acts as a potent inducer of proliferation via activation of the Yorkie oncogene. Reducing CP levels direct apoptosis of overgrowing Src64B-overexpressing tissues. Conversely, overexpressing capping protein inhibits Src64B and Rho1, but not Rac1-induced JNK signaling. CP requires the actin-binding domain of the α-subunit to limit Src64B-induced apoptosis, arguing that the control of F-actin mediates this effect. In turn, JNK directs F-actin accumulation. Moreover, overexpressing capping protein also prevents apoptosis induced by ectopic JNK expression. Our data are consistent with a model in which the control of F-actin by CP limits Src-induced apoptosis or tissue overgrowth by acting downstream of Btk29A, p120ctn and Rho1, but upstream of JNK. In turn, JNK may counteract the effect of CP on F-actin, providing a positive feedback, which amplifies JNK activation. We propose that cytoskeletal changes triggered by misregulation of F-actin modulators may have a significant role in Src-mediated malignant phenotypes during the early stages of cellular transformation.

  3. Particle diffusion from resonance islands in Aladdin at SRC

    Liu, J.; Crosbie, E.; Teng, L.; Bridges, J.; Ciarlette, D.; Kustom, R.; Voss, D.; Mills, F.; Borland, M. [Argonne National Lab., IL (United States); Symon, K. [Univ. of Wisconsin, Madison, WI (US). Dept. of Physics; Trzeciak, W. [Synchrotron Radiation Center, Stoughton, WI (US)


    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.

  4. Dynamic modulation of the Kv2.1 channel by Src-dependent tyrosine phosphorylation

    Song, Min-Young; Hong, Chansik; Bae, Seong Han; So, Insuk; Park, Kang-Sik


    The voltage-gated K+ channel Kv2.1 is expressed as a highly phosphorylated protein in most central neurons, where it plays a key role in regulating neuronal membrane excitability. Previous studies have shown that Kv2.1 channel activity is upregulated by Src-mediated phosphorylation through an unknown mechanism. However, a systematic analysis of the molecular mechanism of Kv2.1 channel phosphorylation by Src is lacking. Here we show that tyrosine phosphorylation by Src plays a fundamental role...

  5. Provenance studies of amber by PIXE

    Lowe, L.; Ruvalcaba S, J.L. [Centro de Estudios Mayas, Instituto de Investigaciones Filologicas, UNAM C.P. 04510 Mexico D.F. (Mexico)


    Analyses by Infrared Spectroscopy and {sup 13} C Nuclear Magnetic Resonance are suitable to determine the paleobotanic source of amber, but cannot differentiate between beds of the same paleobotanic source. Particle Induced X-ray Emission (PIXE) using an external set-up is presented as a new and non-destructive semiquantitative method for provenance studies of amber. PIXE analysis is focused at inorganic contents of amber, considering that the composition of microscopic inclusions depends on the sedimentation environment and it can be used to determine similarities and differences between amber samples and correlate them with amber beds. Results of analyses on amber samples from several world regions and a group of archaeological samples from Chiapas, Mexico, are presented. Amber from different regions have specific inorganic elemental contents; archaeological samples can be associated with beds, even if they have the same paleobotanic origin. (Author)

  6. Giant CBM Field Proven in Shanxi

    Li Mingzhai; Yang Jian


    @@ Qinshui Coalbed Methane Field, the largest one in China, was recently found in the southern part of Qinshui basin in Shanxi Province. Based on the appraisal by experts organized by Ministry of State Land and Resources, the field has a CBM-bearing area of 164.2 square kilometers with the proven geological reserves of 40.210 billion cubic meters and the recoverable reserves of 21.839 billion cubic meters. The field can establish a production capacity for 1 billion cubic meters per year. The CBM field has a stable distribution of CBM layers and abundant reserves. Currently,a total of 29 well have been drilled, of which the highest single-well production is 16000 cubic meters per day. The tested well group is under trial production.

  7. Confocal Microscopy in Biopsy Proven Argyrosis

    Melis Palamar


    Full Text Available Purpose. To evaluate the confocal microscopy findings of a 46-year-old male with bilateral biopsy proven argyrosis. Materials and Methods. Besides routine ophthalmologic examination, anterior segment photography and confocal microscopy with cornea Rostoch module attached to HRT II (Heidelberg Engineering GmbH, Heidelberg, Germany were performed. Findings. Squamous metaplastic changes on conjunctival epithelium and intense highly reflective extracellular punctiform deposits in conjunctival substantia propria were detected. Corneal epithelium was normal. Highly reflective punctiform deposits starting from anterior to mid-stroma and increasing through Descemet’s membrane were evident. Corneal endothelium could not be evaluated due to intense stromal deposits. Conclusion. Confocal microscopy not only supports diagnosis in ocular argyrosis, but also demonstrates the intensity of the deposition in these patients.

  8. A unified framework for managing provenance information in translational research

    Sahoo Satya S


    Full Text Available Abstract Background A critical aspect of the NIH Translational Research roadmap, which seeks to accelerate the delivery of "bench-side" discoveries to patient's "bedside," is the management of the provenance metadata that keeps track of the origin and history of data resources as they traverse the path from the bench to the bedside and back. A comprehensive provenance framework is essential for researchers to verify the quality of data, reproduce scientific results published in peer-reviewed literature, validate scientific process, and associate trust value with data and results. Traditional approaches to provenance management have focused on only partial sections of the translational research life cycle and they do not incorporate "domain semantics", which is essential to support domain-specific querying and analysis by scientists. Results We identify a common set of challenges in managing provenance information across the pre-publication and post-publication phases of data in the translational research lifecycle. We define the semantic provenance framework (SPF, underpinned by the Provenir upper-level provenance ontology, to address these challenges in the four stages of provenance metadata: (a Provenance collection - during data generation (b Provenance representation - to support interoperability, reasoning, and incorporate domain semantics (c Provenance storage and propagation - to allow efficient storage and seamless propagation of provenance as the data is transferred across applications (d Provenance query - to support queries with increasing complexity over large data size and also support knowledge discovery applications We apply the SPF to two exemplar translational research projects, namely the Semantic Problem Solving Environment for Trypanosoma cruzi (T.cruzi SPSE and the Biomedical Knowledge Repository (BKR project, to demonstrate its effectiveness. Conclusions The SPF provides a unified framework to effectively manage provenance

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

    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: [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: [Department of Molecular Cell Biology, Graduate School of Pharmaceutical Sciences, Chiba University, Inohana 1-8-1, Chuo-ku, Chiba 260-8675 (Japan)


    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.

  10. Variability of Physiological Parameters of European Beech Provenances in International Provenance Trials in Serbia

    STOJNIĆ, Srdjan


    Full Text Available In this study, the variability of physiological parameters of five provenances of Europeanbeech (Fagus sylvatica, which were planted at two locations with different ecological conditions atFruška Gora and Debeli Lug, was estimated. Provenance trials were established in the framework ofCOST Action E52: "Evaluation of Beech Genetic Resources for Sustainable Forestry". 2-3 years oldseedlings originating from Croatia, Germany, Bosnia, Austria and Serbia were planted in blocks offifty plants with a spacing of 2 x 1 m. Physiological parameters such as net photosynthesis, rate oftranspiration and stomatal conductance were measured with a portable gas analysis system. Generally,provenances from Fruška Gora Mountain showed higher intensity of all physiological parameters thanprovenances located at site Debeli Lug. High correlations among rates of net photosynthesis andtranspiration, on one side, and stomatal conductance, on the other side, were found. ANOVA testindicates that variability of net photosynthesis, transpiration and stomatal conductance of investigatedprovenances, at the two locations, was influenced both by environmental conditions of sites andgenetic constitution of provenances.

  11. Growth and quality of Grevillea robusta provenances in Ruhande ...


    evaluate the performance of seven provenances and one landrace of G. robusta in terms of ... Key words: Grevillea robusta, land race, provenance, agroecosystems, ..... natural populations and attribute this to be a result of very small founder.

  12. Scientific Workflows + Provenance = Better (Meta-)Data Management

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


    The origin and processing history of an artifact is known as its provenance. Data provenance is an important form of metadata that explains how a particular data product came about, e.g., how and when it was derived in a computational process, which parameter settings and input data were used, etc. Provenance information provides transparency and helps to explain and interpret data products. Other common uses and applications of provenance include quality control, data curation, result debugging, and more generally, 'reproducible science'. Scientific workflow systems (e.g. Kepler, Taverna, VisTrails, and others) provide controlled environments for developing computational pipelines with built-in provenance support. Workflow results can then be explained in terms of workflow steps, parameter settings, input data, etc. using provenance that is automatically captured by the system. Scientific workflows themselves provide a user-friendly abstraction of the computational process and are thus a form of ('prospective') provenance in their own right. The full potential of provenance information is realized when combining workflow-level information (prospective provenance) with trace-level information (retrospective provenance). To this end, the DataONE Provenance Working Group (ProvWG) has developed an extension of the W3C PROV standard, called D-PROV. Whereas PROV provides a 'least common denominator' for exchanging and integrating provenance information, D-PROV adds new 'observables' that described workflow-level information (e.g., the functional steps in a pipeline), as well as workflow-specific trace-level information ( timestamps for each workflow step executed, the inputs and outputs used, etc.) Using examples, we will demonstrate how the combination of prospective and retrospective provenance provides added value in managing scientific data. The DataONE ProvWG is also developing tools based on D-PROV that allow scientists to get more mileage from provenance metadata

  13. Expression, purification, and bioactivity of GST-fused v-Src from a bacterial expression system

    GONG Xing-guo; JI Jing; XIE Jie; ZHOU Yuan; ZHANG Jun-yan; ZHONG Wen-tao


    v-Src is a non-receptor protein tyrosine kinase involved in many signal transduction pathways and closely related to the activation and development of cancers. We present herethe expression, purification, and bioactivity of a GST (glutathione S-transferase)-fused v-Src from a bacterial expression system. Different culture conditions were examined in an isopropyl β-D-thiogalactopyranoside (IPTG)-regulated expression, and the fused protein was purified using GSH (glutathione) affinity chromatography. ELISA (enzyme-linked immunosorbent assay) was employed to determine the phosphorylation kinase activity of the GST-fused v-Src. This strategy seems to be more promising than the insect cell system or other eukaryotic systems employed in earlier Src expression.

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

    Siddiqui Ruqaiyyah


    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

  15. Src Family Kinases and Receptors: Analysis of Three Activation Mechanisms by Dynamic Systems Modeling

    Fuß, Hendrik; Dubitzky, Werner; Downes, C. Stephen; Kurth, Mary Jo


    Src family kinases (SFKs) interact with a number of cellular receptors. They participate in diverse signaling pathways and cellular functions. Most of the receptors involved in SFK signaling are characterized by similar modes of regulation. This computational study discusses a general kinetic model of SFK-receptor interaction. The analysis of the model reveals three major ways of SFK activation: release of inhibition by C-terminal Src kinase, weakening of the inhibitory intramolecular phospho...

  16. Short Rotation Coppice (SRC) Plantations Provide Additional Habitats for Vascular Plant Species in Agricultural Mosaic Landscapes

    BAUM, Sarah; Bolte, Andreas; Weih, Martin


    Increasing loss of biodiversity in agricultural landscapes is often debated in the bioenergy context, especially with respect to non-traditional crops that can be grown for energy production in the future. As promising renewable energy source and additional landscape element, the potential role of short rotation coppice (SRC) plantations to biodiversity is of great interest. We studied plant species richness in eight landscapes (225 km2) containing willow and poplar SRC plantations (1,600 m2)...

  17. Solvent Refined Coal-II (SRC-II) detailed environmental plan


    This document describes environmental research which will: aid in the development of an environmentally acceptable SRC-II process; and provide data for environmental assessment of the process. The SRC-II process is described, criteria for selection of samples to undergo environmental analyses are given, and approximate timelines are presented for obtaining pertinent samples. At this time, the SRC-II process is at the pilot-plant stage of development and a demonstration facility is scheduled to begin operation in 1984. Since design criteria may change, the environmental research described in this document is organized in four phases which correlate with and will provide information early in process development. Phase I research (screening) evaluates samples from existing SRC-II facilities (pilot, process demonstration unit (PDU), bench) which may bracket potential demonstration/commercial practice in terms of physical and chemical criteria. The samples are being subjected to a battery of short-term biomedical and ecological assays. Chemical fractionation and analysis are being performed to determine compounds and compound classes of potential concern. Phase II (baseline) research will evaluate SRC-II materials which are considered most representative of potential demonstration/commercial practice. These materials will be subjected to longer-term, more-extensive biological and ecological analyses relative to effects and environmental fate. Phase III research will examine effects of process modification, control technologies and changing operational conditions on potential environmental properties of SRC-II materials. Phase IV research (onsite monitoring) will develop methods and initiate environmental monitoring for effects at the SRC-II demonstration facility and potential commercial sites. This document also describes industrial hygiene programs which must occur throughout SRC-II process development.

  18. Do Src Kinase and Caveolin Interact Directly with Na,K-ATPase?

    Yosef, Eliyahu; Katz, Adriana; Peleg, Yoav; Mehlman, Tevie; Karlish, Steven J D


    Much evidence points to a role of Na,K-ATPase in ouabain-dependent signal transduction. Based on experiments with different cell lines and native tissue membranes, a current hypothesis postulates direct interactions between the Na,K-ATPase and Src kinase (non-receptor tyrosine kinase). Na,K-ATPase is proposed to bind Src kinase and inhibit its activity, whereas ouabain, the specific Na,K-ATPase inhibitor, binds and stabilizes the E2 conformation, thus exposing the Src kinase domain and its active site Tyr-418 for activation. Ouabain-dependent signaling is thought to be mediated within caveolae by a complex consisting of Na,K-ATPase, caveolin, and Src kinase. In the current work, we have looked for direct interactions utilizing purified recombinant Na,K-ATPase (human α1β1FXYD1 or porcine α1D369Nβ1FXYD1) and purified human Src kinase and human caveolin 1 or interactions between these proteins in native membrane vesicles isolated from rabbit kidney. By several independent criteria and techniques, no stable interactions were detected between Na,K-ATPase and purified Src kinase. Na,K-ATPase was found to be a substrate for Src kinase phosphorylation at Tyr-144. Clear evidence for a direct interaction between purified human Na,K-ATPase and human caveolin was obtained, albeit with a low molar stoichiometry (1:15-30 caveolin 1/Na,K-ATPase). In native renal membranes, a specific caveolin 14-5 oligomer (95 kDa) was found to be in direct interaction with Na,K-ATPase. We inferred that a small fraction of the renal Na,K-ATPase molecules is in a ∼1:1 complex with a caveolin 14-5 oligomer. Thus, overall, whereas a direct caveolin 1/Na,K-ATPase interaction is confirmed, the lack of direct Src kinase/Na,K-ATPase binding requires reassessment of the mechanism of ouabain-dependent signaling.

  19. Effects of gestrinone on uterine leiomyoma and expression of c-Src in a guinea pig model

    Yah ZHU; Xiao-yan QIU; Lei WANG; Jian-hui WU; Gui-ming LIU; Gui-lin HE; Xiu-rong JIANG; Zu-yue SUN; Lin CAO


    Aim: To investigate the effect of gestrinone on uterine leiomyomas and the ex-pression of c-Src, estradiol receptors (ER), and progesterone receptors (PR) in a guinea pig model. Methods: After being oophorectomized, the guinea pigs were allocated into random groups. The model group was treated with estradiol ben-zoate (E2) for 16 weeks. In the gestrinone-treated groups, the animals were treated with E2 for 6 weeks in advance, and then in combination with gestrinone for 10 weeks. Histological examination was performed to evaluate whether there were leiomyoma features in the animals. The protein levels of c-Src, phospho-416Src,ER, and PR were assayed by Western blotting and an immunohistochemical method.Results: Morphological changes were observed in the myometrium of the guinea pig model, including an increase of uterine weights, proliferation of uterine smooth muscles, and the formation of nodules. High protein levels of c-Src, phospho-4'6Src, ER, and PR were observed in the myometrium of the guinea pig model. In the gestrinone-treated group, there were no nodules observed. The histological features of the myometrium were similar to that of the control group. Low protein levels of c-Src, phospho-416Src, ER, and PR were observed in the gestrinone-treated group. Conclusion: The upregulation of c-Src and phospho-416Src indi-cated that the activity of c-Src is augmented in the uterine leiomyoma model, c-Src was associated with the formation of uterine leiomyomas in the model, and gestrinone markedly suppressed the growth of uterine leiomyomas in the model.Gestrinone inhibited not only the protein expression of ER and PR, but also c-Src and the autophosphorylation of c-Src in the guinea pig leiomyoma model.

  20. Genetic and Environmental Models of Circadian Disruption Link SRC-2 Function to Hepatic Pathology.

    Fleet, Tiffany; Stashi, Erin; Zhu, Bokai; Rajapakshe, Kimal; Marcelo, Kathrina L; Kettner, Nicole M; Gorman, Blythe K; Coarfa, Cristian; Fu, Loning; O'Malley, Bert W; York, Brian


    Circadian rhythmicity is a fundamental process that synchronizes behavioral cues with metabolic homeostasis. Disruption of daily cycles due to jet lag or shift work results in severe physiological consequences including advanced aging, metabolic syndrome, and even cancer. Our understanding of the molecular clock, which is regulated by intricate positive feedforward and negative feedback loops, has expanded to include an important metabolic transcriptional coregulator, Steroid Receptor Coactivator-2 (SRC-2), that regulates both the central clock of the suprachiasmatic nucleus (SCN) and peripheral clocks including the liver. We hypothesized that an environmental uncoupling of the light-dark phases, termed chronic circadian disruption (CCD), would lead to pathology similar to the genetic circadian disruption observed with loss of SRC-2 We found that CCD and ablation of SRC-2 in mice led to a common comorbidity of metabolic syndrome also found in humans with circadian disruption, non-alcoholic fatty liver disease (NAFLD). The combination of SRC-2(-/-) and CCD results in a more robust phenotype that correlates with human non-alcoholic steatohepatitis (NASH) and hepatocellular carcinoma (HCC) gene signatures. Either CCD or SRC-2 ablation produces an advanced aging phenotype leading to increased mortality consistent with other circadian mutant mouse models. Collectively, our studies demonstrate that SRC-2 provides an essential link between the behavioral activities influenced by light cues and the metabolic homeostasis maintained by the liver.

  1. Depletion of insulin receptor substrate 2 reverses oncogenic transformation induced by v-src

    Hong-zhi SUN; Lin XU; Bo ZHOU; Wei-jin ZANG; Shu-fang WU


    Aim: To investigate the role of insulin receptor substrate 2 (IRS-2) in oncogenic transformation induced by v-src. Methods: IRS-2 gene was silenced using small interfering RNAs (siRNAs). Nuclear translocation and interaction of IRS-2 with v-src was determined using subcellular fractionation, confocal microscopy, and immunoprecipitation. The activity of the cyclin D1 promoter and r-DNA promoter was measured with a luciferase assay.Results: Depletion of IRS-2 inhibited R-/v-src cell growth and reverse the oncogenic transformation. IRS-2 bound to src via its two PI3-K binding sites, which are critical for activities involved in the transformation. Nuclear IRS-2 occupied the cyclin D1 and rDNA promoters. The combination of IRS-2 and v-src increased the activity of the two promoters, especially the rDNA promoter.Conclusion: Depletion of insulin receptor substrate 2 could reverse oncogenic transformation induced by v-src.

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

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


    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.

  3. SRC-2 Is an Essential Coactivator for Orchestrating Metabolism and Circadian Rhythm

    Erin Stashi


    Full Text Available Synchrony of the mammalian circadian clock is achieved by complex transcriptional and translational feedback loops centered on the BMAL1:CLOCK heterodimer. Modulation of circadian feedback loops is essential for maintaining rhythmicity, yet the role of transcriptional coactivators in driving BMAL1:CLOCK transcriptional networks is largely unexplored. Here, we show diurnal hepatic steroid receptor coactivator 2 (SRC-2 recruitment to the genome that extensively overlaps with the BMAL1 cistrome during the light phase, targeting genes that enrich for circadian and metabolic processes. Notably, SRC-2 ablation impairs wheel-running behavior, alters circadian gene expression in several peripheral tissues, alters the rhythmicity of the hepatic metabolome, and deregulates the synchronization of cell-autonomous metabolites. We identify SRC-2 as a potent coregulator of BMAL1:CLOCK and find that SRC-2 targets itself with BMAL1:CLOCK in a feedforward loop. Collectively, our data suggest that SRC-2 is a transcriptional coactivator of the BMAL1:CLOCK oscillators and establish SRC-2 as a critical positive regulator of the mammalian circadian clock.

  4. Cell invasion by Neisseria meningitidis requires a functional interplay between the focal adhesion kinase, Src and cortactin.

    Heiko Slanina

    Full Text Available Entry of Neisseria meningitidis (the meningococcus into human brain microvascular endothelial cells (HBMEC is mediated by fibronectin or vitronectin bound to the surface protein Opc forming a bridge to the respective integrins. This interaction leads to cytoskeletal rearrangement and uptake of meningococci. In this study, we determined that the focal adhesion kinase (FAK, which directly associates with integrins, is involved in integrin-mediated internalization of N. meningitidis in HBMEC. Inhibition of FAK activity by the specific FAK inhibitor PF 573882 reduced Opc-mediated invasion of HBMEC more than 90%. Moreover, overexpression of FAK mutants that were either impaired in the kinase activity or were not capable of autophosphorylation or overexpression of the dominant-negative version of FAK (FRNK blocked integrin-mediated internalization of N. meningitidis. Importantly, FAK-deficient fibroblasts were significantly less invaded by N. meningitidis. Furthermore, N. meningitidis induced tyrosine phosphorylation of several host proteins including the FAK/Src complex substrate cortactin. Inhibition of cortactin expression by siRNA silencing and mutation of critical amino acid residues within cortactin, that encompass Arp2/3 association and dynamin binding, significantly reduced meningococcal invasion into eukaryotic cells suggesting that both domains are critical for efficient uptake of N. meningitidis into eukaryotic cells. Together, these results indicate that N. meningitidis exploits the integrin signal pathway for its entry and that FAK mediates the transfer of signals from activated integrins to the cytoskeleton. A cooperative interplay between FAK, Src and cortactin then enables endocytosis of N. meningitidis into host cells.

  5. Tracking Provenance in ORNL's Flexible Research Platforms

    Hensley, Zachary P [ORNL; Sanyal, Jibonananda [ORNL; New, Joshua Ryan [ORNL


    Provenance is dened as information about the origin of objects, a concept that applies to both physical and digital objects and often overlaps both. The use of provenance in systems designed for research is an important but forgotten feature. Provenance allows for proper and exact tracking of information, its use, its lineage, its derivations and other metadata that are important for correctly adhering to the scien- tic method. In our project's prescribed use of provenance, researchers can determine detailed information about the use of sensor data in their experiments on ORNL's Flexible Research Platforms (FRPs). Our project's provenance system, Provenance Data Management System (ProvDMS), tracks information starting with the creation of information by an FRP sensor. The system determines station information, sensor information, and sensor channel information. The system allows researchers to derive generations of experiments from the sensor data and tracks their hierarchical flow. Key points can be seen in the history of the information as part of the information's workflow. The concept of provenance and its usage in science is relatively new and while used in other cases around the world, our project's provenance diers in a key area. To keep track of provenance, most systems must be designed or redesigned around the new provenance system. Our system is designed as a cohesive but sepa- rate entity and allows for researchers to continue using their own methods of analysis without being constrained in their ways in order to track the provenance. We have designed ProvDMS using a lightweight provenance library, Core Provenance Library (CPL) v.6 In addition to keeping track of sensor data experiments and its provenance, ProvDMS also provides a web-enabled visualization of the inheritance.

  6. Data Provenance Architecture for the Geosciences

    Murphy, F.; Irving, D. H.


    The pace at which geoscientific insights inform societal development quickens with time and these insights drive decisions and actions of ever-increasing human and economic significance. Until recently academic, commercial and government bodies have maintained distinct bodies of knowledge to support scientific enquiry as well as societal development. However, it has become clear that the curation of the body of data is an activity of equal or higher social and commercial value. We address the community challenges in the curation of, access to, and analysis of scientific data including: the tensions between creators, providers and users; incentives and barriers to sharing; ownership and crediting. We also discuss the technical and financial challenges in maximising the return on the effort made in generating geoscientific data. To illustrate how these challenges might be addressed in the broader geoscientific domain, we describe the high-level data governance and analytical architecture in the upstream Oil Industry. This domain is heavily dependent on costly and highly diverse geodatasets collected and assimilated over timeframes varying from seconds to decades. These data must support both operational decisions at the minute-hour timefame and strategic and economic decisions of enterprise or national scale, and yet be sufficiently robust to last the life of a producing field. We develop three themes around data provenance, data ownership and business models for data curation. 1/ The overarching aspiration is to ensure that data provenance and quality is maintained along the analytical workflow. Hence if data on which a publication or report changes, the report and its publishers can be notified and we describe a mechanism by which dependent knowledge products can be flagged. 2/ From a cost and management point of view we look at who "owns" data especially in cases where the cost of curation and stewardship is significant compared to the cost of acquiring the data

  7. Src-like adaptor protein 2 (SLAP2) binds to and inhibits FLT3 signaling

    Moharram, Sausan A.; Chougule, Rohit A.; Su, Xianwei; Li, Tianfeng; Sun, Jianmin; Zhao, Hui; Rönnstrand, Lars; Kazi, Julhash U.


    Fms-like tyrosine kinase (FLT3) is a frequently mutated oncogene in acute myeloid leukemia (AML). FLT3 inhibitors display promising results in a clinical setting, but patients relapse after short-term treatment due to the development of resistant disease. Therefore, a better understanding of FLT3 downstream signal transduction pathways will help to identify an alternative target for the treatment of AML patients carrying oncogenic FLT3. Activation of FLT3 results in phosphorylation of FLT3 on several tyrosine residues that recruit SH2 domain-containing signaling proteins. We screened a panel of SH2 domain-containing proteins and identified SLAP2 as a potent interacting partner of FLT3. We demonstrated that interaction occurs when FLT3 is activated, and also, an intact SH2 domain of SLAP2 is required for binding. SLAP2 binding sites in FLT3 mainly overlap with those of SRC. SLAP2 over expression in murine proB cells or myeloid cells inhibited oncogenic FLT3-ITD-mediated cell proliferation and colony formation in vitro, and tumor formation in vivo. Microarray analysis suggests that higher SLAP2 expression correlates with a gene signature similar to that of loss of oncogene function. Furthermore, FLT3-ITD positive AML patients with higher SLAP2 expression displayed better prognosis compared to those with lower expression of SLAP2. Expression of SLAP2 blocked FLT3 downstream signaling cascades including AKT, ERK, p38 and STAT5. Finally, SLAP2 accelerated FLT3 degradation through enhanced ubiquitination. Collectively, our data suggest that SLAP2 acts as a negative regulator of FLT3 signaling and therefore, modulation of SLAP2 expression levels may provide an alternative therapeutic approach for FLT3-ITD positive AML. PMID:27458164

  8. Revised SRC-I project baseline. Volume 2


    The SRC Process Area Design Baseline consists of six volumes. The first four were submitted to DOE on 9 September 1981. The fifth volume, summarizing the Category A Engineering Change Proposals (ECPs), was not submitted. The sixth volume, containing proprietary information on Kerr-McGee's Critical Solvent Deashing System, was forwarded to BRHG Synthetic Fuels, Inc. for custody, according to past instructions from DOE, and is available for perusal by authorized DOE representatives. DOE formally accepted the Design Baseline under ICRC Release ECP 4-1001, at the Project Configuration Control Board meeting in Oak Ridge, Tennessee on 5 November 1981. The documentation was then revised by Catalytic, Inc. to incorporate the Category B and C and Post-Baseline Engineering Change Proposals. Volumes I through V of the Revised Design Baseline, dated 22 October 1982, are nonproprietary and they were issued to the DOE via Engineering Change Notice (ECN) 4-1 on 23 February 1983. Volume VI again contains proprieary information on Kerr-McGee Critical Solvent Deashing System; it was issued to Burns and Roe Synthetic Fuels, Inc. Subsequently, updated process descriptions, utility summaries, and errata sheets were issued to the DOE and Burns and Roe Synthetic Fuels, Inc. on nonproprietary Engineering Change Notices 4-2 and 4-3 on 24 May 1983.

  9. Data provenance assurance in the cloud using blockchain

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


    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.

  10. Automatic Provenance Recording for Scientific Data using Trident

    Simmhan, Y.; Barga, R.; van Ingen, C.


    Provenance is increasingly recognized as being critical to the understanding and reuse of scientific datasets. Given the rapid generation of scientific data from sensors and computational model results, it is not practical to manually record provenance for data and automated techniques for provenance capture are essential. Scientific workflows provide a framework for representing computational models and complex transformations of scientific data, and present a means for tracking the operations performed to derive a dataset. The Trident Scientific Workbench is a workflow system that natively incorporates provenance capture of data derived as part of the workflow execution. The applications used as part of a Trident workflow can execute on a remote computational cluster, such as a supercomputing center on in the Cloud, or on the local desktop of the researcher, and provenance on data derived by the applications is seamlessly captured. Scientists also have the option to annotate the provenance metadata using domain specific tags such as, for example, GCMD keywords. The provenance records thus captured can be exported in the Open Provenance Model* XML format that is emerging as a provenance standard in the eScience community or visualized as a graph of data and applications. The Trident workflow system and provenance recorded by it has been successfully applied in the Neptune oceanography project and is presently being tested in the Pan-STARRS astronomy project. *

  11. Short amino acid stretches can mediate amyloid formation in globular proteins: the Src homology 3 (SH3) case.

    Ventura, Salvador; Zurdo, Jesús; Narayanan, Saravanakumar; Parreño, Matilde; Mangues, Ramón; Reif, Bernd; Chiti, Fabrizio; Giannoni, Elisa; Dobson, Christopher M; Aviles, Francesc X; Serrano, Luis


    Protein misfolding and deposition underlie an increasing number of debilitating human disorders. We have shown that model proteins unrelated to disease, such as the Src homology 3 (SH3) domain of the p58alpha subunit of bovine phosphatidyl-inositol-3'-kinase (PI3-SH3), can be converted in vitro into assemblies with structural and cytotoxic properties similar to those of pathological aggregates. By contrast, homologous proteins, such as alpha-spectrin-SH3, lack the capability of forming amyloid fibrils at a measurable rate under any of the conditions we have so far examined. However, transplanting a small sequence stretch (6 aa) from PI3-SH3 to alpha-spectrin-SH3, comprising residues of the diverging turn and adjacent RT loop, creates an amyloidogenic protein closely similar in its behavior to the original PI3-SH3. Analysis of specific PI3-SH3 mutants further confirms the involvement of this region in conferring amyloidogenic properties to this domain. Moreover, the inclusion in this stretch of two consensus residues favored in SH3 sequences substantially inhibits aggregation. These findings show that short specific amino acid stretches can act as mediators or facilitators in the incorporation of globular proteins into amyloid structures, and they support the suggestion that natural protein sequences have evolved in part to code for structural characteristics other than those included in the native fold, such as avoidance of aggregation.

  12. v-Src-induced nuclear localization of YAP is involved in multipolar spindle formation in tetraploid cells.

    Kakae, Keiko; Ikeuchi, Masayoshi; Kuga, Takahisa; Saito, Youhei; Yamaguchi, Naoto; Nakayama, Yuji


    The protein-tyrosine kinase, c-Src, is involved in a variety of signaling events, including cell division. We have reported that v-Src, which is a mutant variant of the cellular proto-oncogene, c-Src, causes delocalization of Aurora B kinase, resulting in a furrow regression in cytokinesis and the generation of multinucleated cells. However, the effect of v-Src on mitotic spindle formation is unknown. Here we show that v-Src-expressing HCT116 and NIH3T3 cells undergo abnormal cell division, in which cells separate into more than two cells. Upon v-Src expression, the proportion of multinucleated cells is increased in a time-dependent manner. Flow cytometry analysis revealed that v-Src increases the number of cells having a ≥4N DNA content. Microscopic analysis showed that v-Src induces the formation of multipolar spindles with excess centrosomes. These results suggest that v-Src induces multipolar spindle formation by generating multinucleated cells. Tetraploidy activates the tetraploidy checkpoint, leading to a cell cycle arrest of tetraploid cells at the G1 phase, in which the nuclear exclusion of the transcription co-activator YAP plays a critical role. In multinucleated cells that are induced by cytochalasin B and the Plk1 inhibitor, YAP is excluded from the nucleus. However, v-Src prevents this nuclear exclusion of YAP through a decrease in the phosphorylation of YAP at Ser127 in multinucleated cells. Furthermore, v-Src decreases the expression level of p53, which also plays a critical role in the cell cycle arrest of tetraploid cells. These results suggest that v-Src promotes abnormal spindle formation in at least two ways: generation of multinucleated cells and a weakening of the tetraploidy checkpoint. Copyright © 2016 Elsevier Inc. All rights reserved.

  13. The Cbl Proto-Oncogene Product Negatively Regulates the Src-Family Tyrosine Kinase Fyn by Enhancing Its Degradation


    Fyn is a prototype Src-family tyrosine kinase that plays specific roles in neural development, keratinocyte differentiation, and lymphocyte activation, as well as roles redundant with other Src-family kinases. Similar to other Src-family kinases, efficient regulation of Fyn is achieved through intramolecular binding of its SH3 and SH2 domains to conserved regulatory regions. We have investigated the possibility that the tyrosine kinase regulatory protein Cbl provides a complementary mechanism...

  14. Everything is autoimmune until proven otherwise.

    Shoenfeld, Yehuda


    diseases that have either an inflammatory and/or specific autoimmune response, we have to keep an open eye because everything is potentially autoimmune until proven otherwise.

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

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


    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

  16. Protein-tyrosine Phosphatase and Kinase Specificity in Regulation of SRC and Breast Tumor Kinase* ♦

    Fan, Gaofeng; Aleem, Saadat; Yang, Ming; Miller, W. Todd; Tonks, Nicholas K.


    Despite significant evidence to the contrary, the view that phosphatases are “nonspecific” still pervades the field. Systems biology approaches to defining how signal transduction pathways are integrated at the level of whole organisms also often downplay the contribution of phosphatases, defining them as “erasers” that serve merely to restore the system to its basal state. Here, we present a study that counteracts the idea of “nonspecific phosphatases.” We have characterized two structurally similar and functionally related kinases, BRK and SRC, which are regulated by combinations of activating autophosphorylation and inhibitory C-terminal sites of tyrosine phosphorylation. We demonstrated specificity at the level of the kinases in that SRMS phosphorylated the C terminus of BRK, but not SRC; in contrast, CSK is the kinase responsible for C-terminal phosphorylation of SRC, but not BRK. For the phosphatases, we observed that RNAi-mediated suppression of PTP1B resulted in opposing effects on the activity of BRK and SRC and have defined the mechanisms underlying this specificity. PTP1B inhibited BRK by directly dephosphorylating the Tyr-342 autophosphorylation site. In contrast, PTP1B potentiated SRC activity, but not by dephosphorylating SRC itself directly; instead, PTP1B regulated the interaction between CBP/PAG and CSK. SRC associated with, and phosphorylated, the transmembrane protein CBP/PAG at Tyr-317, resulting in CSK recruitment. We identified PAG as a substrate of PTP1B, and dephosphorylation abolished recruitment of the inhibitory kinase CSK. Overall, these findings illustrate how the combinatorial effects of PTKs and PTPs may be integrated to regulate signaling, with both classes of enzymes displaying exquisite specificity. PMID:25897081

  17. Protein-tyrosine Phosphatase and Kinase Specificity in Regulation of SRC and Breast Tumor Kinase.

    Fan, Gaofeng; Aleem, Saadat; Yang, Ming; Miller, W Todd; Tonks, Nicholas K


    Despite significant evidence to the contrary, the view that phosphatases are "nonspecific" still pervades the field. Systems biology approaches to defining how signal transduction pathways are integrated at the level of whole organisms also often downplay the contribution of phosphatases, defining them as "erasers" that serve merely to restore the system to its basal state. Here, we present a study that counteracts the idea of "nonspecific phosphatases." We have characterized two structurally similar and functionally related kinases, BRK and SRC, which are regulated by combinations of activating autophosphorylation and inhibitory C-terminal sites of tyrosine phosphorylation. We demonstrated specificity at the level of the kinases in that SRMS phosphorylated the C terminus of BRK, but not SRC; in contrast, CSK is the kinase responsible for C-terminal phosphorylation of SRC, but not BRK. For the phosphatases, we observed that RNAi-mediated suppression of PTP1B resulted in opposing effects on the activity of BRK and SRC and have defined the mechanisms underlying this specificity. PTP1B inhibited BRK by directly dephosphorylating the Tyr-342 autophosphorylation site. In contrast, PTP1B potentiated SRC activity, but not by dephosphorylating SRC itself directly; instead, PTP1B regulated the interaction between CBP/PAG and CSK. SRC associated with, and phosphorylated, the transmembrane protein CBP/PAG at Tyr-317, resulting in CSK recruitment. We identified PAG as a substrate of PTP1B, and dephosphorylation abolished recruitment of the inhibitory kinase CSK. Overall, these findings illustrate how the combinatorial effects of PTKs and PTPs may be integrated to regulate signaling, with both classes of enzymes displaying exquisite specificity. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  18. CFTR impairment upregulates c-Src activity through IL-1β autocrine signaling.

    Massip-Copiz, María Macarena; Clauzure, Mariángeles; Valdivieso, Ángel Gabriel; Santa-Coloma, Tomás Antonio


    Cystic Fibrosis (CF) is a disease caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. Previously, we found several genes showing a differential expression in CFDE cells (epithelial cells derived from a CF patient). One corresponded to c-Src; its expression and activity was found increased in CFDE cells, acting as a signaling molecule between the CFTR activity and MUC1 overexpression. Here we report that bronchial IB3-1 cells (CF cells) also showed increased c-Src activity compared to 'CFTR-corrected' S9 cells. In addition, three different Caco-2 cell lines, each stably transfected with a different CFTR-specific shRNAs, displayed increased c-Src activity. The IL-1β receptor antagonist IL1RN reduced the c-Src activity of Caco-2/pRS26 cells (expressing a CFTR-specific shRNA). In addition, increased mitochondrial and cellular ROS levels were detected in Caco-2/pRS26 cells. ROS levels were partially reduced by incubation with PP2 (c-Src inhibitor) or IL1RN, and further reduced by using the NOX1/4 inhibitor GKT137831. Thus, IL-1β→c-Src and IL-1β→NOX signaling pathways appear to be responsible for the production of cellular and mitochondrial ROS in CFTR-KD cells. In conclusion, IL-1β constitutes a new step in the CFTR signaling pathway, located upstream of c-Src, which is stimulated in cells with impaired CFTR activity.

  19. Analysis of the c-src gene product structure, abundance, and protein kinase activity in human neuroblastoma and glioblastoma cells.

    O'Shaughnessy, J; Deseau, V; Amini, S; Rosen, N; Bolen, J B


    We have compared in different human neuroblastoma cell lines and human glioblastoma cells the expression level, structure, and tyrosine-specific protein kinase activity of pp60c-src. Our results show that not all human neuroblastoma cell lines express pp60c-src molecules with amino-terminal structural alterations. In neuroblastoma cells which possess pp60c-src with altered gel migration, the diminished polyacrylamide gel mobility of pp60c-src was found not to be dependent upon amino-terminal phosphorylations since extensive treatment of these molecules with phosphatase did not significantly change their gel migration properties. Similar differences in gel migration were observed when RNA from the various neuroblastoma and glioblastoma cells was translated in vitro using either rabbit reticulocyte or wheat germ lysates. White the level of c-src mRNA in the different cells analyzed was found to be similar, the abundance of pp60c-src in these same cells was found to vary by as much as 12-fold. This suggests that the abundance of pp60c-src in human neuroendocrine tumors is regulated through post-transcriptional and/or post-translational events which may be related to the stage of neuronal differentiation of the cells. Based upon determination of pp60c-src abundance by immunoblot analysis, we demonstrate that pp60c-src molecules derived from human neuroblastoma and glioblastoma cells have very similar in vitro protein kinase activities.

  20. c-Src Inhibition Improves Cardiovascular Function but not Remodeling or Fibrosis in Angiotensin II-Induced Hypertension.

    Callera, Glaucia E; Antunes, Tayze T; He, Ying; Montezano, Augusto C; Yogi, Alvaro; Savoia, Carmine; Touyz, Rhian M


    c-Src plays an important role in angiotensin II (Ang II) signaling. Whether this member of the Src family kinases is involved in the development of Ang II-induced hypertension and associated cardiovascular damage in vivo remains unknown. Here, we studied Ang II-infused (400 ng/kg/min) mice in which c-Src was partially deleted (c-Src(+/-)) and in wild-type (WT, c-Src(+/+)) mice treated with a c-Src inhibitor (CGP077675; 25 mg/kg/d). Ang II increased blood pressure and induced endothelial dysfunction in WT mice, responses that were ameliorated in c-Src(+/-) and CGP077675-treated mice. Vascular wall thickness and cross-sectional area were similarly increased by Ang II in WT and c-Src(+/-) mice. CGP077675 further increased cross-sectional area in hypertensive mice. Cardiac dysfunction (ejection fraction and fractional shortening) in Ang II-infused WT mice was normalized in c-Src(+/-) mice. Increased oxidative stress (plasma thiobarbituric acid-reactive substances, hydrogen peroxide, and vascular superoxide generation) in Ang II-infused WT mice was attenuated in c-Src-deficient and CGP077675-treated mice. Hyperactivation of vascular c-Src, ERK1/2 (extracellular signal-regulated kinase 1/2), and JNK (c-Jun N-terminal kinase) in hypertensive mice was normalized in CGP077675-treated and c-Src(+/-) mice. Vascular fibronectin was increased by Ang II in all groups and further augmented by CGP077675. Cardiac fibrosis and inflammation induced by Ang II were amplified in c-Src(+/-) and CGP-treated mice. Our data indicate that although c-Src downregulation attenuates development of hypertension, improves endothelial and cardiac function, reduces oxidative stress, and normalizes vascular signaling, it has little beneficial effect on fibrosis. These findings suggest a divergent role for c-Src in Ang II-dependent hypertension, where c-Src may be more important in regulating redox-sensitive cardiac and vascular function than fibrosis and remodeling. © 2016 American Heart Association

  1. Src1 is a Protein of the Inner Nuclear Membrane Interacting with the Dictyostelium Lamin NE81

    Petros Batsios


    Full Text Available The nuclear envelope (NE consists of the outer and inner nuclear membrane (INM, whereby the latter is bound to the nuclear lamina. Src1 is a Dictyostelium homologue of the helix-extension-helix family of proteins, which also includes the human lamin-binding protein MAN1. Both endogenous Src1 and GFP-Src1 are localized to the NE during the entire cell cycle. Immuno-electron microscopy and light microscopy after differential detergent treatment indicated that Src1 resides in the INM. FRAP experiments with GFP-Src1 cells suggested that at least a fraction of the protein could be stably engaged in forming the nuclear lamina together with the Dictyostelium lamin NE81. Both a BioID proximity assay and mis-localization of soluble, truncated mRFP-Src1 at cytosolic clusters consisting of an intentionally mis-localized mutant of GFP-NE81 confirmed an interaction of Src1 and NE81. Expression GFP-Src11–646, a fragment C-terminally truncated after the first transmembrane domain, disrupted interaction of nuclear membranes with the nuclear lamina, as cells formed protrusions of the NE that were dependent on cytoskeletal pulling forces. Protrusions were dependent on intact microtubules but not actin filaments. Our results indicate that Src1 is required for integrity of the NE and highlight Dictyostelium as a promising model for the evolution of nuclear architecture.

  2. Data Provenance and Data Management in eScience

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


    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...

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

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


    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.

  4. Src family kinase inhibitor PP2 efficiently inhibits cervical cancer cell proliferation through down-regulating phospho-Src-Y416 and phospho-EGFR-Y1173.

    Kong, Lu; Deng, Zhihong; Shen, Haiying; Zhang, Yuxiang


    Tyrosine (Y) kinases inhibitors have been approved for targeted treatment of cancer. However, their clinical use is limited to some cancers and the mechanism of their action remains unclear. Previous study has indicated that PP2, a selective inhibitor of the Src family of non-receptor tyrosine kinases (nRTK), efficiently repressed cervical cancer growth in vitro and in vivo. In this regard, our aims are to explore the mechanism of PP2 on cervical cancer cell growth inhibition by investigating the suppressive divergence among PP1, PP2, and a negative control compound PP3. MTT results showed that three compounds had different inhibitory effects on proliferation of two cervical cancer cells, HeLa and SiHa, and PP2 was most efficient in a time- and dose-dependent manner. Moreover, we found 10 μM PP2 down-regulated pSrc-Y416 (P < 0.05), pEGFR-Y845 (P < 0.05), and -Y1173 (P < 0.05) expression levels, while 10 μM PP1 down-regulated pSrc-Y416 (P < 0.05) and pEGFR-Y845 (P < 0.05), but not pEGFR-Y1173; 10 μM PP3 down-regulated only pEGFR-Y1173 (P < 0.05). PP2 could modulate cell cycle arrest by up-regulating p21(Cip1) and p27(Kip1) in both HeLa and SiHa cells and down-regulating expression of cyclin A, and cyclin dependent kinase-2, -4 (Cdk-2, -4) in HeLa and of cyclin B and Cdk-2 in SiHa. Our results indicate that Src pathway and EGFR pathway play different roles in the proliferation of cervical cancer cells and PP2 efficiently reduces cervical cancer cell proliferation by reduction of both Src and EGFR activity.

  5. Defining the specificity space of the human SRC homology 2 domain.

    Huang, Haiming; Li, Lei; Wu, Chenggang; Schibli, David; Colwill, Karen; Ma, Sucan; Li, Chengjun; Roy, Protiva; Ho, Krystina; Songyang, Zhou; Pawson, Tony; Gao, Youhe; Li, Shawn S-C


    Src homology 2 (SH2) domains are the largest family of interaction modules encoded by the human genome to recognize tyrosine-phosphorylated sequences and thereby play pivotal roles in transducing and controlling cellular signals emanating from protein-tyrosine kinases. Different SH2 domains select for distinct phosphopeptides, and the function of a given SH2 domain is often dictated by the specific motifs that it recognizes. Therefore, deciphering the phosphotyrosyl peptide motif recognized by an SH2 domain is the key to understanding its cellular function. Here we cloned all 120 SH2 domains identified in the human genome and determined the phosphotyrosyl peptide binding properties of 76 SH2 domains by screening an oriented peptide array library. Of these 76, we defined the selectivity for 43 SH2 domains and refined the binding motifs for another 33 SH2 domains. We identified a number of novel binding motifs, which are exemplified by the BRDG1 SH2 domain that selects specifically for a bulky, hydrophobic residue at P + 4 relative to the Tyr(P) residue. Based on the oriented peptide array library data, we developed scoring matrix-assisted ligand identification (or SMALI), a Web-based program for predicting binding partners for SH2-containing proteins. When applied to SH2D1A/SAP (SLAM-associated protein), a protein whose mutation or deletion underlies the X-linked lymphoproliferative syndrome, SMALI not only recapitulated known interactions but also identified a number of novel interacting proteins for this disease-associated protein. SMALI also identified a number of potential interactors for BRDG1, a protein whose function is largely unknown. Peptide in-solution binding analysis demonstrated that a SMALI score correlates well with the binding energy of a peptide to a given SH2 domain. The definition of the specificity space of the human SH2 domain provides both the necessary molecular basis and a platform for future exploration of the functions for SH2-containing

  6. Numerical Simulation of Response of SRC Columns Subjected to Blast Loading

    SUN Jianyun; LI Guoqiang; LU Yong


    The dynamic characteristics and failure modes of steel reinforced concrete (SRC) columns subjected to blast loading are complicated because of the transient stress wave in the SRC columns and the interaction between steel and concrete.This paper presents a numerical simulation of the response of SRC columns subjected to blast loading using hydrocode LS-DYNA.In the numerical model,a sophisticate concrete material model (the Concrete Damage Model) is employed with consideration of the strain rate effect and the damage accumulation.An erosion technique is adopted to model the spalling process of concrete.The possible failure modes of SRC columns are evaluated.It is observed that the failure of SRC columns subjected to blast load can generally be classified into three modes,namely,a direct failure in concrete body due to the stress wave,a transverse shear failure near the support sections due to the high shear force,and a flexural failure pertaining to large local and global deformation of the reinforcing steel.

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

    Görtzen, Jan; Schierwagen, Robert; Bierwolf, Jeanette


    . 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....... RESULTS: Transcription of albumin and RhoA was decreased, whereas transcription and activation of c-SRC was increased in hepatocytes cultured on 12 kPa compared to 1 kPa gels. LX2 cells cultured on 12 kPa 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 12 kPa gels. In LX2 cells with RhoA overexpression, c-SRC inhibition by PP2 failed to improve fibrosis. RhoA expression was significantly elevated in human and experimental liver fibrosis, while c-SRC was inactivated. CONCLUSIONS...

  8. A Src-like inactive conformation in the abl tyrosine kinase domain.

    Nicholas M Levinson


    Full Text Available The improper activation of the Abl tyrosine kinase results in chronic myeloid leukemia (CML. The recognition of an inactive conformation of Abl, in which a catalytically important Asp-Phe-Gly (DFG motif is flipped by approximately 180 degrees with respect to the active conformation, underlies the specificity of the cancer drug imatinib, which is used to treat CML. The DFG motif is not flipped in crystal structures of inactive forms of the closely related Src kinases, and imatinib does not inhibit c-Src. We present a structure of the kinase domain of Abl, determined in complex with an ATP-peptide conjugate, in which the protein adopts an inactive conformation that resembles closely that of the Src kinases. An interesting aspect of the Src-like inactive structure, suggested by molecular dynamics simulations and additional crystal structures, is the presence of features that might facilitate the flip of the DFG motif by providing room for the phenylalanine to move and by coordinating the aspartate side chain as it leaves the active site. One class of mutations in BCR-Abl that confers resistance to imatinib appears more likely to destabilize the inactive Src-like conformation than the active or imatinib-bound conformations. Our results suggest that interconversion between distinctly different inactive conformations is a characteristic feature of the Abl kinase domain.

  9. Seismic cyclic loading test of SRC columns confined with 5-spirals

    WENG ChengChiang; YIN YenLiang; WANG JuiChen; LIANG ChingYu


    Presented herein is an experimental study on seismic resistance of rectangular steel reinforced concrete (SRC) columns confined with a new type of multi-spiral cage.The multi-spiral cage is a device of five interconnected spirals,named "5-spirals",with a large spiral at the center and four small ones at the corners.The innovation of applying the 5-spirals to SRC column is to take its superiority in concrete confinement and efficiency in automatic production for the precast con-struction industry.Four full-scale SRC columns were tested under horizontal cyclic loading.All of the tested columns were capable of sustaining a drift angle up to 6%radians.The hysteresis loops observed from the cyclic loading tests indicated that the spirally confined SRC columns demonstrated excellent performances in both strength and ductility.The test results suggested that with significant saving of the confinement steel,the newly innovated 5-spirals can be successfully applied to the precast rectangular SRC columns.

  10. Fyn and Src are Effectors of Oncogenic EGFR Signaling in Glioblastoma Patients

    Lu, Kan V.; Zhu, Shaojun; Cvrljevic, Anna; Huang, Tiffany T.; Sarkaria, Shawn; Ahkavan, David; Dang, Julie; Dinca, Eduard B.; Plaisier, Seema B.; Oderberg, Isaac; Lee, Yohan; Chen, Zugen; Caldwell, Jeremy S.; Xie, Yongmin; Loo, Joseph A.; Seligson, David; Chakravari, Arnab; Lee, Francis Y.; Weinmann, Roberto; Cloughesy, Timothy F.; Nelson, Stanley F.; Bergers, Gabriele; Graeber, Thomas; Furnari, Frank B.; James, C. David; Cavenee, Webster K.; Johns, Terrance G.; Mischel, Paul S.


    Activating EGFR mutations are common in many cancers including glioblastoma. However, clinical responses to EGFR inhibitors are infrequent and short-lived. We demonstrate that the Src family kinases (SFKs) Fyn and Src are effectors of oncogenic EGFR signaling, enhancing invasion and tumor cell survival in vivo. Expression of a constitutively active EGFR mutant, EGFRvIII, resulted in activating phosphorylation and physical association with Src and Fyn, promoting tumor growth and motility. Gene silencing of Fyn and Src limited EGFR and EGFRvIII-dependent tumor cell motility. The SFK inhibitor dasatinib inhibited invasion, promoted tumor regression and induced apoptosis in vivo, significantly prolonging survival of an orthotopic glioblastoma model expressing endogenous EGFRvIII. Dasatinib enhanced the efficacy of an anti-EGFR monoclonal antibody (mAb 806) in vivo, further limiting tumor growth and extending survival. Examination of a large cohort of clinical samples demonstrated frequent coactivation of EGFR and SFKs in glioblastoma patients. These results establish a mechanism linking EGFR signaling with Fyn and Src activation to promote tumor progression and invasion in vivo and provide rationale for combined anti-EGFR and anti-SFK targeted therapies. PMID:19690143

  11. Cigarette Smoke Activates the Proto-Oncogene c-Src to Promote Airway Inflammation and Lung Tissue Destruction

    Geraghty, Patrick; Hardigan, Andrew


    The diagnosis of chronic obstructive pulmonary disease (COPD) confers a 2-fold increased lung cancer risk even after adjusting for cigarette smoking, suggesting that common pathways are operative in both diseases. Although the role of the tyrosine kinase c-Src is established in lung cancer, less is known about its impact in other lung diseases, such as COPD. This study examined whether c-Src activation by cigarette smoke contributes to the pathogenesis of COPD. Cigarette smoke increased c-Src activity in human small airway epithelial (SAE) cells from healthy donors and in the lungs of exposed mice. Similarly, higher c-Src activation was measured in SAE cells from patients with COPD compared with healthy control subjects. In SAE cells, c-Src silencing or chemical inhibition prevented epidermal growth factor (EGF) receptor signaling in response to cigarette smoke but not EGF stimulation. Further studies showed that cigarette smoke acted through protein kinase C α to trigger c-Src to phosphorylate EGF receptor and thereby to induce mitogen-activated protein kinase responses in these cells. To further investigate the role of c-Src, A/J mice were orally administered the specific Src inhibitor AZD-0530 while they were exposed to cigarette smoke for 2 months. AZD-0530 treatment blocked c-Src activation, decreased macrophage influx, and prevented airspace enlargement in the lungs of cigarette smoke–exposed mice. Moreover, inhibiting Src deterred the cigarette smoke–mediated induction of matrix metalloproteinase-9 and -12 in alveolar macrophages and lung expression of cathepsin K, IL-17, TNF-α, MCP-1, and KC, all key factors in the pathogenesis of COPD. These results indicate that activation of the proto-oncogene c-Src by cigarette smoke promotes processes linked to the development of COPD. PMID:24111605

  12. Adaptation of eastern whitepine provenances to planting sites

    Maurice E., Jr. Demeritt; Peter W. Garrett


    Eastern white pine provenances from the extreme limits of the natural range of this species are changing from above- and below-average stability to average stability for height growth with increasing age. The regression method is useful for evaluating the stability of provenance to planting sites. The same general conclusions are reached for the performance at...

  13. Research Traceability using Provenance Services for Biomedical Analysis

    Anjum, Ashiq; Branson, Andrew; Habib, Irfan; McClatchey, Richard; Solomonides, Tony


    We outline the approach being developed in the neuGRID project to use provenance management techniques for the purposes of capturing and preserving the provenance data that emerges in the specification and execution of workflows in biomedical analyses. In the neuGRID project a provenance service has been designed and implemented that is intended to capture, store, retrieve and reconstruct the workflow information needed to facilitate users in conducting user analyses. We describe the architecture of the neuGRID provenance service and discuss how the CRISTAL system from CERN is being adapted to address the requirements of the project and then consider how a generalised approach for provenance management could emerge for more generic application to the (Health)Grid community.

  14. Research traceability using provenance services for biomedical analysis.

    Anjum, Ashiq; Bloodsworth, Peter; Branson, Andrew; Habib, Irfan; McClatchey, Richard; Solomonides, Tony; Soomro, Kamran; The Neugrid Consortium


    We outline the approach being developed in the neuGRID project to use provenance management techniques for the purposes of capturing and preserving the provenance data that emerges in the specification and execution of workflows in biomedical analyses. In the neuGRID project a provenance service has been designed and implemented that is intended to capture, store, retrieve and reconstruct the workflow information needed to facilitate users in conducting user analyses. We describe the architecture of the neuGRID provenance service and discuss how the CRISTAL system from CERN is being adapted to address the requirements of the project and then consider how a generalised approach for provenance management could emerge for more generic application to the (Health)Grid community.

  15. Src activation by β-adrenoreceptors is a key switch for tumour metastasis.

    Armaiz-Pena, Guillermo N; Allen, Julie K; Cruz, Anthony; Stone, Rebecca L; Nick, Alpa M; Lin, Yvonne G; Han, Liz Y; Mangala, Lingegowda S; Villares, Gabriel J; Vivas-Mejia, Pablo; Rodriguez-Aguayo, Cristian; Nagaraja, Archana S; Gharpure, Kshipra M; Wu, Zheng; English, Robert D; Soman, Kizhake V; Shahzad, Mian M K; Shazhad, Mian M K; Zigler, Maya; Deavers, Michael T; Zien, Alexander; Soldatos, Theodoros G; Jackson, David B; Wiktorowicz, John E; Torres-Lugo, Madeline; Young, Tom; De Geest, Koen; Gallick, Gary E; Bar-Eli, Menashe; Lopez-Berestein, Gabriel; Cole, Steve W; Lopez, Gustavo E; Lutgendorf, Susan K; Sood, Anil K


    Noradrenaline can modulate multiple cellular functions important for cancer progression; however, how this single extracellular signal regulates such a broad array of cellular processes is unknown. Here we identify Src as a key regulator of phosphoproteomic signalling networks activated in response to beta-adrenergic signalling in cancer cells. These results also identify a new mechanism of Src phosphorylation that mediates beta-adrenergic/PKA regulation of downstream networks, thereby enhancing tumour cell migration, invasion and growth. In human ovarian cancer samples, high tumoural noradrenaline levels were correlated with high pSrc(Y419) levels. Moreover, among cancer patients, the use of beta blockers was significantly associated with reduced cancer-related mortality. Collectively, these data provide a pivotal molecular target for disrupting neural signalling in the tumour microenvironment.

  16. Role of SRC-like adaptor protein (SLAP) in immune and malignant cell signaling.

    Kazi, Julhash U; Kabir, Nuzhat N; Rönnstrand, Lars


    SRC-like adaptor protein (SLAP) is an adaptor protein structurally similar to the SRC family protein kinases. Like SRC, SLAP contains an SH3 domain followed by an SH2 domain but the kinase domain has been replaced by a unique C-terminal region. SLAP is expressed in a variety of cell types. Current studies suggest that it regulates signaling of various cell surface receptors including the B cell receptor, the T cell receptor, cytokine receptors and receptor tyrosine kinases which are important regulator of immune and cancer cell signaling. SLAP targets receptors, or its associated components, by recruiting the ubiquitin machinery and thereby destabilizing signaling. SLAP directs receptors to ubiquitination-mediated degradation and controls receptors turnover as well as signaling. Thus, SLAP appears to be an important component in regulating signal transduction required for immune and malignant cells.

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

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


    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.

  18. c-Src Kinase Inhibition Reduces Arrhythmia Inducibility and Connexin43 Dysregulation after Myocardial Infarction

    Rutledge, Cody A.; Ng, Fu Siong; Sulkin, Matthew S.; Greener, Ian D.; Sergeyenko, Artem M.; Liu, Hong; Gemel, Joanna; Beyer, Eric C.; Sovari, Ali A.; Efimov, Igor R.; Dudley, Samuel C.


    Objectives The aim of this study was to evaluate the role of c-Src inhibition on connexin43 (Cx43) regulation in a mouse model of myocardial infarction (MI). Background MI is associated with decreased expression of Cx43, the principal gap junction protein responsible for propagating current in ventricles. Activated c-Src has been linked to Cx43 dysregulation. Methods MI was induced in 12-week-old mice by coronary artery occlusion. MI mice were treated with c-Src inhibitors (PP1 or AZD0530), PP3 (an inactive analogue of PP1), or saline. Treated hearts were compared to sham mice by echocardiography, optical mapping, telemetry ECG monitoring, and inducibility studies. Tissues were collected for immunoblotting, quantitative PCR, and immunohistochemistry. Results Active c-Src was elevated in PP3-treated MI mice compared to sham at the scar border (280%, p=0.003) and distal ventricle (346%, p=0.013). PP1 treatment restored active c-Src to sham levels at the scar border (86%, p=0.95) and distal ventricle (94%, p=1.0). PP1 raised Cx43 expression by 69% in the scar border (p=0.048) and by 73% in distal ventricle (p=0.043) compared to PP3 mice. PP1-treated mice had restored conduction velocity at the scar border (PP3: 32 cm/s, PP1: 41 cm/s, p < 0.05) and lower arrhythmic inducibility (PP3: 71%, PP1: 35%, p < 0.05) than PP3 mice. PP1 did not change infarct size, ECG pattern, or cardiac function. AZD0530 treatment demonstrated restoration of Cx43 comparable to PP1. Conclusions c-Src inhibition improved Cx43 levels and conduction velocity and lowered arrhythmia inducibility after MI, suggesting a new approach for arrhythmia reduction following MI. PMID:24361364

  19. The Activation of c-Src Tyrosine Kinase: Conformational Transition Pathway and Free Energy Landscape.

    Fajer, Mikolai; Meng, Yilin; Roux, Benoît


    Tyrosine kinases are important cellular signaling allosteric enzymes that regulate cell growth, proliferation, metabolism, differentiation, and migration. Their activity must be tightly controlled, and malfunction can lead to a variety of diseases, particularly cancer. The nonreceptor tyrosine kinase c-Src, a prototypical model system and a representative member of the Src-family, functions as complex multidomain allosteric molecular switches comprising SH2 and SH3 domains modulating the activity of the catalytic domain. The broad picture of self-inhibition of c-Src via the SH2 and SH3 regulatory domains is well characterized from a structural point of view, but a detailed molecular mechanism understanding is nonetheless still lacking. Here, we use advanced computational methods based on all-atom molecular dynamics simulations with explicit solvent to advance our understanding of kinase activation. To elucidate the mechanism of regulation and self-inhibition, we have computed the pathway and the free energy landscapes for the "inactive-to-active" conformational transition of c-Src for different configurations of the SH2 and SH3 domains. Using the isolated c-Src catalytic domain as a baseline for comparison, it is observed that the SH2 and SH3 domains, depending upon their bound orientation, promote either the inactive or active state of the catalytic domain. The regulatory structural information from the SH2-SH3 tandem is allosterically transmitted via the N-terminal linker of the catalytic domain. Analysis of the conformational transition pathways also illustrates the importance of the conserved tryptophan 260 in activating c-Src, and reveals a series of concerted events during the activation process.

  20. Cardiomyocyte apoptosis triggered by RAFTK/pyk2 via Src kinase is antagonized by paxillin.

    Melendez, Jaime; Turner, Christopher; Avraham, Hava; Steinberg, Susan F; Schaefer, Erik; Sussman, Mark A


    Altered cellular adhesion and apoptotic signaling in cardiac remodeling requires coordinated regulation of multiple constituent proteins that comprise cytoskeletal focal adhesions. One such protein activated by cardiac remodeling is related adhesion focal tyrosine kinase (RAFTK, also known as pyk2). Adenoviral-mediated expression of RAFTK in neonatal rat cardiomyocytes involves concurrent increases in phosphorylation of Src, c-Jun N-terminal kinase, and p38 leading to characteristic apoptotic changes including cleavage of poly(ADP-ribose) polymerase, caspase-3 activation, and increased DNA laddering. DNA laddering was decreased by mutation of the Tyr(402) Src-binding site in RAFTK, suggesting a central role for Src activity in apoptotic cell death that was confirmed by adenoviral-mediated Src expression. Multiple apoptotic signaling cascades are recruited by RAFTK as demonstrated by prevention of apoptosis using caspase-3 inhibitor IV (caspase-3 specific inhibitor), PP2 (Src-specific kinase inhibitor), or Csk (cellular negative regulator for Src), as well as dominant negative constructs for p38beta or MKP-1. These RAFTK-mediated phenotypic characteristics are prevented by concurrent expression of wild-type or a phosphorylation-deficient paxillin mutated at Tyr(31) and Tyr(118). Wild-type or mutant paxillin protein accumulation in the cytoplasm has no overt effect upon cell structure, but paxillin accumulation prevents losses of myofibril organization as well as focal adhesion kinase, vinculin, and paxillin protein levels mediated by RAFTK. Apoptotic signaling cascade inhibition by paxillin indicates interruption of signaling proximal to but downstream of RAFTK activity. Chronic RAFTK activation in cardiac remodeling may represent a maladaptive reactive response that can be modulated by paxillin, opening up novel possibilities for inhibition of cardiomyocyte apoptosis and structural degeneration in heart failure.

  1. c-Src Links a RANK/αvβ3 Integrin Complex to the Osteoclast Cytoskeleton

    Izawa, Takashi; Zou, Wei; Chappel, Jean C.; Ashley, Jason W.; Feng, Xu


    RANK ligand (RANKL), by mechanisms unknown, directly activates osteoclasts to resorb bone. Because c-Src is key to organizing the cell's cytoskeleton, we asked if the tyrosine kinase also mediates RANKL-stimulated osteoclast activity. RANKL induces c-Src to associate with RANK369–373 in an αvβ3-dependent manner. Furthermore, RANK369–373 is the only one of six putative TRAF binding motifs sufficient to generate actin rings and activate the same cytoskeleton-organizing proteins as the integrin. While c-Src organizes the cell's cytoskeleton in response to the cytokine, it does not participate in RANKL-stimulated osteoclast formation. Attesting to their collaboration, αvβ3 and activated RANK coprecipitate, but only in the presence of c-Src. c-Src binds activated RANK via its Src homology 2 (SH2) domain and αvβ3 via its SH3 domain, suggesting the kinase links the two receptors. Supporting this hypothesis, deletion or inactivating point mutation of either the c-Src SH2 or SH3 domain obviates the RANK/αvβ3 association. Thus, activated RANK prompts two distinct signaling pathways; one promotes osteoclast formation, and the other, in collaboration with c-Src-mediated linkage to αvβ3, organizes the cell's cytoskeleton. PMID:22615494

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

    Wang Jing


    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.

  3. Receptor protein tyrosine phosphatase alpha activates Src-family kinases and controls integrin-mediated responses in fibroblasts

    Su, J; Muranjan, M; Sap, J


    BACKGROUND: Fyn and c-Src are two of the most widely expressed Src-family kinases. Both are strongly implicated in the control of cytoskeletal organization and in the generation of integrin-dependent signalling responses in fibroblasts. These proteins are representative of a large family of tyros...

  4. Blockage of Src by Specific siRNA as a Novel Therapeutic Strategy to Prevent Destructive Repair in Steroid-Associated Osteonecrosis in Rabbits.

    Zheng, Li-zhen; Cao, Hui-juan; Chen, Shi-hui; Tang, Tao; Fu, Wei-min; Huang, Le; Chow, Dick Ho Kiu; Wang, Yi-xiang; Griffith, James Francis; He, Wei; Zhou, Hong; Zhao, De-wei; Zhang, Ge; Wang, Xin-luan; Qin, Ling


    Vascular hyperpermeability and highly upregulated bone resorption in the destructive repair progress of steroid-associated osteonecrosis (SAON) are associated with a high expression of VEGF and high Src activity (Src is encoded by the cellular sarcoma [c-src] gene). This study was designed to prove our hypothesis that blocking the VEGF-Src signaling pathway by specific Src siRNA is able to prevent destructive repair in a SAON rabbit model. Destructive repair in SAON was induced in rabbits. At 2, 4, and 6 weeks after SAON induction, VEGF, anti-VEGF, Src siRNA, Src siRNA+VEGF, control siRNA, and saline were introduced via intramedullary injection into proximal femora for each group, respectively. Vascularization and permeability were quantified by dynamic contrast-enhanced (DCE) MRI. At week 6 after SAON induction, proximal femurs were dissected for micro-computed tomography (μCT)-based trabecular architecture with finite element analysis (FEA), μCT-based angiography, and histological analysis. Histological evaluation revealed that VEGF enhanced destructive repair, whereas anti-VEGF prevented destructive repair and Src siRNA and Src siRNA+VEGF prevented destructive repair and enhanced reparative osteogenesis. Findings of angiography and histomorphometry were consistent with those determined by DCE MRI. Src siRNA inhibited VEGF-mediated vascular hyperpermeability but preserved VEGF-induced neovascularization. Bone resorption was enhanced in the VEGF group and inhibited in the anti-VEGF, Src siRNA, Src siRNA+VEGF groups as determined by both 3D μCT and 2D histomorphometry. FEA showed higher estimated failure load in the Src siRNA and Src siRNA+VEGF groups when compared to the vehicle control group. Blockage of VEGF-Src signaling pathway by specific Src siRNA was able to prevent steroid-associated destructive repair while improving reconstructive repair in SAON, which might become a novel therapeutic strategy.

  5. Meeting at mitosis: cell cycle-specific regulation of c-Src by RPTPalpha.

    Mustelin, Tomas; Hunter, Tony


    Exquisite regulation is required for cells to properly enter and exit the phases of the cell cycle. The transmembrane receptor-like protein tyrosine phosphatase RPTPalpha, an important protein that participates in the transition of the cell cycle from G2 to mitosis activates the protein tyrosine kinase c-Src in vivo. Mustelin and Hunter discuss new findings that describe the highly regulated activation of RPTPalpha and c-Src that occurs just before entry into the mitotic phase. These findings also raise several questions that pertain to redistribution of RPTPalpha in the cell, and the role of phosphorylation and dimerization in regulating RPTPalpha activity.

  6. Fractionally distilled SRC-I, SRC-II, EDS, H-Coal and ITSL direct coal liquefaction process materials: a comparative summary of chemical analysis and biological testing

    Wright, C.W.; Later, D.W.; Dauble, D.D.; Wilson, B.W.


    This document reports and compares the results compiled from chemical analyses and biological testing of coal liquefaction process materials which were fractionally distilled, after production, into various comparable boiling-point range cuts. Comparative analyses were performed on solvent refined coal (SRC)-I, SRC-II, H-Coal, EDS an integrated two-stage liquefaction (ITSL) distillate materials. Mutagenicity and carcinogenicity assays were conducted in conjunction with chromatographic and mass spectrometric analyses to provide detailed, comparative, chemical and biological assessments. Where possible, results obtained from the distillate cuts are compared to those from coal liquefaction materials with limited boiling ranges. Work reported here was conducted by investigators in the Biology and Chemistry Department at the Pacific Northwest Laboratory (PNL), Richland, WA. 38 refs., 16 figs., 27 tabs.

  7. Komadu: A Capture and Visualization System for Scientific Data Provenance

    Isuru Suriarachchi


    Full Text Available Data provenance captured from scientific applications is a critical precursor to data sharing and reuse. For researchers wanting to repurpose data, it is a source of information about the lineage and attribution of the data and this is needed in order to establish trust in a data set. Komadu is a standalone provenance capture and visualization system for capturing, representing, and manipulating provenance coming from scientific tools, infrastructures, and repositories. It uses the W3C PROV standard [1] in representing data, and it is the successor of the Karma [2] provenance capture system which was based on Open Provenance Model (OPM [3]. Komadu comes with two different interfaces: a Web Services interface based on Apache Axis2 [4] and a messaging interface based on RabbitMQ [5]. Komadu is completely open source and the source code is publicly available on GitHub [6]. Even though Komadu has been used most extensively in relation to scientific research, its interfaces are designed to collect and visualize provenance of any kind of application needing provenance.

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

    Martina Vojtěchová


    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.

  9. Src promotes castration-recurrent prostate cancer through androgen receptor-dependent canonical and non-canonical transcriptional signatures

    Chattopadhyay, Indranil; Wang, Jianmin; Qin, Maochun; Gao, Lingqiu; Holtz, Renae; Vessella, Robert L.; Leach, Robert W.; Gelman, Irwin H.


    Progression of prostate cancer (PC) to castration-recurrent growth (CRPC) remains dependent on sustained expression and transcriptional activity of the androgen receptor (AR). A major mechanism contributing to CRPC progression is through the direct phosphorylation and activation of AR by Src-family (SFK) and ACK1 tyrosine kinases. However, the AR-dependent transcriptional networks activated by Src during CRPC progression have not been elucidated. Here, we show that activated Src (Src527F) induces androgen-independent growth in human LNCaP cells, concomitant with its ability to induce proliferation/survival genes normally induced by dihydrotestosterone (DHT) in androgen-dependent LNCaP and VCaP cells. Src induces additional gene signatures unique to CRPC cell lines, LNCaP-C4-2 and CWR22Rv1, and to CRPC LuCaP35.1 xenografts. By comparing the Src-induced AR-cistrome and/or transcriptome in LNCaP to those in CRPC and LuCaP35.1 tumors, we identified an 11-gene Src-regulated CRPC signature consisting of AR-dependent, AR binding site (ARBS)-associated genes whose expression is altered by DHT in LNCaP[Src527F] but not in LNCaP cells. The differential expression of a subset (DPP4, BCAT1, CNTNAP4, CDH3) correlates with earlier PC metastasis onset and poorer survival, with the expression of BCAT1 required for Src-induced androgen-independent proliferation. Lastly, Src enhances AR binding to non-canonical ARBS enriched for FOXO1, TOP2B and ZNF217 binding motifs; cooperative AR/TOP2B binding to a non-canonical ARBS was both Src- and DHT-sensitive and correlated with increased levels of Src-induced phosphotyrosyl-TOP2B. These data suggest that CRPC progression is facilitated via Src-induced sensitization of AR to intracrine androgen levels, resulting in the engagement of canonical and non-canonical ARBS-dependent gene signatures. PMID:28055971

  10. The Symbiotic Relationship between Scientific Workflow and Provenance (Invited)

    Stephan, E.


    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

  11. South Polar Residual Cap Geomorphology and Inferred Environmental Changes

    Byrne, S.; Ingersoll, A.; Pathare, A.


    The CO2 southern residual cap (SRC) both controls circulation patterns regionally and buffers the atmospheric pressure globally. In turn this CO2 deposit is affected by changes in environmental conditions wrought by external forces such as dust storm activity. Mars Global Surveyor data of this area have revealed a rich variety of geomorphic features (1) of which there are several distinct classes. These different classes may be end members of the same basic process of insolation driven ablation. We are currently investigating two types of SRC features. Swiss-cheese features (SCF) are depressions characterized by flat floors and steep walls, which retreat 1-3 meters each Martian year (2). In some regions they have a definite symmetry axis along the north-south direction (3). After the seasonal frost disappears the residual ice exposed in the walls has a lower albedo (4). Previously (5) we modeled the evolution and growth of these depressions as a hole in a layer of CO2 ice underlain by water ice, which best explains their morphologic and thermal properties. The observed thickness of the CO2 slab can be as high as 8 meters but in general is much lower. Larger SCF?s commonly possess a raised central island of CO2 surrounded by a moat that penetrates to the underlying water ice (3). The fast rate of wall retreat observed (2) combined with the small sizes of the SCF?s indicate that all SCF?s visible today were created geologically recently. Within a particular region the size distribution is quite narrow (3): no larger (older) or smaller (younger) features were seen indicating that some relatively abrupt change in environmental conditions initiated the growth of this particular population of features. Fingerprint terrain (1) are areas with evenly spaced parallel ridges, which are steeper on one side. These ridges may have small areas of water ice exposed in the intervening troughs. Their wavelength is on the order of 70-90m with the steep edges facing northeast although

  12. DataONE: A Data Federation with Provenance Support

    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


    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.

  13. A model of provenance applied to biodiversity datasets

    Amanqui, Flor K; De Nies, Tom; Dimou, Anastasia; Verborgh, Ruben; Mannens, Erik; Van De Walle, Rik; Moreira, Dilvan


    Nowadays, the Web has become one of the main sources of biodiversity information. An increasing number of biodiversity research institutions add new specimens and their related information to their biological collections and make this information available on the Web. However, mechanisms which are currently available provide insufficient provenance of biodiversity information. In this paper, we propose a new biodiversity provenance model extending the W3C PROV Data Model. Biodiversity data is...

  14. 76 FR 44605 - Alaska Region's Subsistence Resource Commission (SRC) Program; Public Meeting and Teleconference


    ... National Interest Lands Conservation Act, Public Law 96-487, to operate in accordance with the provisions... National Park SRC Agenda 1. Call to order. 2. Welcome and Introductions. 3. Administrative Announcements. 4.... ] 11. Old Business: a. Nabesna Off-Road Vehicle Management Plan Final Environmental Impact Statement. b...

  15. Solvent refined coal (SRC) process. Annual technical progress report, January 1979-December 1979


    A set of statistically designed experiments was used to study the effects of several important operating variables on coal liquefaction product yield structures. These studies used a Continuous Stirred-Tank Reactor to provide a hydrodynamically well-defined system from which kinetic data could be extracted. An analysis of the data shows that product yield structures can be adequately represented by a correlative model. It was shown that second-order effects (interaction and squared terms) are necessary to provide a good model fit of the data throughout the range studied. Three reports were issued covering the SRC-II database and yields as functions of operating variables. The results agree well with the generally-held concepts of the SRC reaction process, i.e., liquid phase hydrogenolysis of liquid coal which is time-dependent, thermally activated, catalyzed by recycle ash, and reaction rate-controlled. Four reports were issued summarizing the comprehensive SRC reactor thermal response models and reporting the results of several studies made with the models. Analytical equipment for measuring SRC off-gas composition and simulated distillation of coal liquids and appropriate procedures have been established.

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

    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.

  17. An evolutionary switch in ND2 enables Src kinase regulation of NMDA receptors.

    Scanlon, David P; Bah, Alaji; Krzeminski, Mickaël; Zhang, Wenbo; Leduc-Pessah, Heather L; Dong, Yi Na; Forman-Kay, Julie D; Salter, Michael W


    The non-receptor tyrosine kinase Src is a key signalling hub for upregulating the function of N-methyl D-aspartate receptors (NMDARs). Src is anchored within the NMDAR complex via NADH dehydrogenase subunit 2 (ND2), a mitochondrially encoded adaptor protein. The interacting regions between Src and ND2 have been broadly identified, but the interaction between ND2 and the NMDAR has remained elusive. Here we generate a homology model of ND2 and dock it onto the NMDAR via the transmembrane domain of GluN1. This interaction is enabled by the evolutionary loss of three helices in bilaterian ND2 proteins compared to their ancestral homologues. We experimentally validate our model and demonstrate that blocking this interaction with an ND2 fragment identified in our experimental studies prevents Src-mediated upregulation of NMDAR currents in neurons. Our findings establish the mode of interaction between an NMDAR accessory protein with one of the core subunits of the receptor.

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

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


    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…

  19. Dynamin Forms a Src Kinase–sensitive Complex with Cbl and Regulates Podosomes and Osteoclast Activity

    Bruzzaniti, Angela; Neff, Lynn; Sanjay, Archana; Horne, William C.; De Camilli, Pietro; Baron, Roland


    Podosomes are highly dynamic actin-containing adhesion structures found in osteoclasts, macrophages, and Rous sarcoma virus (RSV)-transformed fibroblasts. After integrin engagement, Pyk2 recruits Src and the adaptor protein Cbl, forming a molecular signaling complex that is critical for cell migration, and deletion of any molecule in this complex disrupts podosome ring formation and/or decreases osteoclast migration. Dynamin, a GTPase essential for endocytosis, is also involved in actin cytoskeleton remodeling and is localized to podosomes where it has a role in actin turnover. We found that dynamin colocalizes with Cbl in the actin-rich podosome belt of osteoclasts and that dynamin forms a complex with Cbl in osteoclasts and when overexpressed in 293VnR or SYF cells. The association of dynamin with Cbl in osteoclasts was decreased by Src tyrosine kinase activity and we found that destabilization of the dynamin-Cbl complex involves the recruitment of Src through the proline-rich domain of Cbl. Overexpression of dynamin increased osteoclast bone resorbing activity and migration, whereas overexpression of dynK44A decreased osteoclast resorption and migration. These studies suggest that dynamin, Cbl, and Src coordinately participate in signaling complexes that are important in the assembly and remodeling of the actin cytoskeleton, leading to changes in osteoclast adhesion, migration, and resorption. PMID:15872089

  20. Dynamin forms a Src kinase-sensitive complex with Cbl and regulates podosomes and osteoclast activity.

    Bruzzaniti, Angela; Neff, Lynn; Sanjay, Archana; Horne, William C; De Camilli, Pietro; Baron, Roland


    Podosomes are highly dynamic actin-containing adhesion structures found in osteoclasts, macrophages, and Rous sarcoma virus (RSV)-transformed fibroblasts. After integrin engagement, Pyk2 recruits Src and the adaptor protein Cbl, forming a molecular signaling complex that is critical for cell migration, and deletion of any molecule in this complex disrupts podosome ring formation and/or decreases osteoclast migration. Dynamin, a GTPase essential for endocytosis, is also involved in actin cytoskeleton remodeling and is localized to podosomes where it has a role in actin turnover. We found that dynamin colocalizes with Cbl in the actin-rich podosome belt of osteoclasts and that dynamin forms a complex with Cbl in osteoclasts and when overexpressed in 293VnR or SYF cells. The association of dynamin with Cbl in osteoclasts was decreased by Src tyrosine kinase activity and we found that destabilization of the dynamin-Cbl complex involves the recruitment of Src through the proline-rich domain of Cbl. Overexpression of dynamin increased osteoclast bone resorbing activity and migration, whereas overexpression of dynK44A decreased osteoclast resorption and migration. These studies suggest that dynamin, Cbl, and Src coordinately participate in signaling complexes that are important in the assembly and remodeling of the actin cytoskeleton, leading to changes in osteoclast adhesion, migration, and resorption.

  1. Drosophila PRL-1 is a growth inhibitor that counteracts the function of the Src oncogene.

    Pagarigan, Krystle T; Bunn, Bryce W; Goodchild, Jake; Rahe, Travis K; Weis, Julie F; Saucedo, Leslie J


    Phosphatase of Regenerating Liver (PRL) family members have emerged as molecular markers that significantly correlate to the ability of many cancers to metastasize. However, contradictory cellular responses to PRL expression have been reported, including the inhibition of cell cycle progression. An obvious culprit for the discrepancy is the use of dozens of different cell lines, including many isolated from tumors or cultured cells selected for immortalization which may have missing or mutated modulators of PRL function. We created transgenic Drosophila to study the effects of PRL overexpression in a genetically controlled, organismal model. Our data support the paradigm that the normal cellular response to high levels of PRL is growth suppression and furthermore, that PRL can counter oncogenic activity of Src. The ability of PRL to inhibit growth under normal conditions is dependent on a CAAX motif that is required to localize PRL to the apical edge of the lateral membrane. However, PRL lacking the CAAX motif can still associate indiscriminately with the plasma membrane and retains its ability to inhibit Src function. We propose that PRL binds to other membrane-localized proteins that are effectors of Src or to Src itself. This first examination of PRL in a model organism demonstrates that PRL performs as a tumor suppressor and underscores the necessity of identifying the conditions that enable it to transform into an oncogene in cancer.

  2. Desmoglein 3, via an interaction with E-cadherin, is associated with activation of Src.

    Siu Man Tsang

    Full Text Available BACKGROUND: Desmoglein 3 (Dsg3, a desmosomal adhesion protein, is expressed in basal and immediate suprabasal layers of skin and across the entire stratified squamous epithelium of oral mucosa. However, increasing evidence suggests that the role of Dsg3 may involve more than just cell-cell adhesion. METHODOLOGY/PRINCIPAL FINDINGS: To determine possible additional roles of Dsg3 during epithelial cell adhesion we used overexpression of full-length human Dsg3 cDNA, and RNAi-mediated knockdown of this molecule in various epithelial cell types. Overexpression of Dsg3 resulted in a reduced level of E-cadherin but a colocalisation with the E-cadherin-catenin complex of the adherens junctions. Concomitantly these transfected cells exhibited marked migratory capacity and the formation of filopodial protrusions. These latter events are consistent with Src activation and, indeed, Src-specific inhibition reversed these phenotypes. Moreover Dsg3 knockdown, which also reversed the decreased level of E-cadherin, partially blocked Src phosphorylation. CONCLUSIONS/SIGNIFICANCE: Our data are consistent with the possibility that Dsg3, as an up-stream regulator of Src activity, helps regulate adherens junction formation.

  3. Fisetin Suppresses Macrophage-Mediated Inflammatory Responses by Blockade of Src and Syk.

    Kim, Jun Ho; Kim, Mi-Yeon; Kim, Jong-Hoon; Cho, Jae Youl


    Flavonoids, such as fisetin (3,7,3',4'-tetrahydroxyflavone), are plant secondary metabolites. It has been reported that fisetin is able to perform numerous pharmacological roles including anti-inflammatory, anti-microbial, and anti-cancer activities; however, the exact anti-inflammatory mechanism of fisetin is not understood. In this study, the pharmacological action modes of fisetin in lipopolysaccharide (LPS)-stimulated macrophage-like cells were elucidated by using immunoblotting analysis, kinase assays, and an overexpression strategy. Fisetin diminished the release of nitric oxide (NO) and reduced the mRNA levels of inducible NO synthase (iNOS), tumor necrosis factor (TNF)-α, and cyclooxygenase (COX)-2 in LPS-stimulated RAW264.7 cells without displaying cytotoxicity. This compound also blocked the nuclear translocation of p65/nuclear factor (NF)-κB. In agreement, the upstream phosphorylation events for NF-κB activation, composed of Src, Syk, and IκBα, were also reduced by fisetin. The phospho-Src level, triggered by overexpression of wild-type Src, was also inhibited by fisetin. Therefore, these results strongly suggest that fisetin can be considered a bioactive immunomodulatory compound with anti-inflammatory properties through suppression of Src and Syk activities.

  4. In silico investigation of potential SRC kinase ligands from traditional Chinese medicine.

    Weng Ieong Tou

    Full Text Available Src kinase is an attractive target for drug development based on its established relationship with cancer and possible link to hypertension. The suitability of traditional Chinese medicine (TCM compounds as potential drug ligands for further biological evaluation was investigated using structure-based, ligand-based, and molecular dynamics (MD analysis. Isopraeroside IV, 9alpha-hydroxyfraxinellone-9-O-beta-D-glucoside (9HFG and aurantiamide were the top three TCM candidates identified from docking. Hydrogen bonds and hydrophobic interactions were the primary forces governing docking stability. Their stability with Src kinase under a dynamic state was further validated through MD and torsion angle analysis. Complexes formed by TCM candidates have lower total energy estimates than the control Sacaratinib. Four quantitative-structural activity relationship (QSAR in silico verifications consistently suggested that the TCM candidates have bioactive properties. Docking conformations of 9HFG and aurantiamide in the Src kinase ATP binding site suggest potential inhibitor-like characteristics, including competitive binding at the ATP binding site (Lys295 and stabilization of the catalytic cleft integrity. The TCM candidates have significantly lower ligand internal energies and are estimated to form more stable complexes with Src kinase than Saracatinib. Structure-based and ligand-based analysis support the drug-like potential of 9HFG and aurantiamide and binding mechanisms reveal the tendency of these two candidates to compete for the ATP binding site.

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

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


    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

  6. Restful Implementation of Catalogue Service for Geospatial Data Provenance

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


    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.

  7. Putting Lipstick on Pig: Enabling Database-style Workflow Provenance

    Amsterdamer, Yael; Deutch, Daniel; Milo, Tova; Stoyanovich, Julia; Tannen, Val


    Workflow provenance typically assumes that each module is a "black-box", so that each output depends on all inputs (coarse-grained dependencies). Furthermore, it does not model the internal state of a module, which can change between repeated executions. In practice, however, an output may depend on only a small subset of the inputs (fine-grained dependencies) as well as on the internal state of the module. We present a novel provenance framework that marries database-style and workflow-style provenance, by using Pig Latin to expose the functionality of modules, thus capturing internal state and fine-grained dependencies. A critical ingredient in our solution is the use of a novel form of provenance graph that models module invocations and yields a compact representation of fine-grained workflow provenance. It also enables a number of novel graph transformation operations, allowing to choose the desired level of granularity in provenance querying (ZoomIn and ZoomOut), and supporting "what-if" workflow analyti...

  8. Lightweight Provenance Service for High-Performance Computing

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


    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.

  9. Human melanoma cells express FGFR/Src/Rho signaling that entails an adhesion-independent caveolin-1 membrane association.

    Fecchi, Katia; Travaglione, Sara; Spadaro, Francesca; Quattrini, Adriano; Parolini, Isabella; Piccaro, Giovanni; Raggi, Carla; Fabbri, Alessia; Felicetti, Federica; Carè, Alessandra; Fiorentini, Carla; Sargiacomo, Massimo


    Caveolae have been indicated as a center of cytoskeleton regulation for Src kinase/Rho GTPase signaling. In addition, Src recruitment on intact cortical actin cytoskeleton appears to be required for bFGF/FGFR signal activation. Recently, we established a relationship between caveolin-1 (Cav-1) expression and cell migration in human malignant melanoma, constitutively activated by a bFGF autoregulatory loop. This work intends to investigate whether caveolae's asset, through bFGF/FGFR/c-Src/Rho signaling, could be related to melanoma cell anchorage. Accordingly, we revealed the existence of a FGFR/Src kinase pathway in Cav-1 enriched detergent-resistant membranes (DRMs) of Me665/1 metastatic melanoma cells, as confirmed by FGFR silencing. Moreover, we determined the expression and phosphorylation levels of Cav-1/Src/Erk signal pathway as a function of FGFR activation and cell density. A sucrose density gradient ultracentrifugation was employed to monitor Cav-1 membrane association and buoyancy in Me665/1 cells treated for actin fragmentation or for altered phosphorylation signals. As a result, melanoma cells show remarkable resistance to Cav-1 disassembly, together with persisting cell signal activity, being Src and Cav-1 crucial modulators of Rho GTPases. In conclusion, our study primarily highlights, in a metastatic melanoma cell line expressing caveolin, the circumstances whereby caveola structural and functional endurance enables the FGFR/Src/Rho GTPases pathway to keep on cell progression.

  10. Differential mitotic activation of endogenous c-Src, c-Yes, and Lyn in HeLa cells.

    Kuga, Takahisa; Nakayama, Yuji; Hoshino, Masaki; Higashiyama, Yukihiro; Obata, Yuuki; Matsuda, Daisuke; Kasahara, Kousuke; Fukumoto, Yasunori; Yamaguchi, Naoto


    Src-family tyrosine kinases (SFKs) play an important role in mitosis. Despite overlapping expression of multiple SFK members, little is known about how individual SFK members are activated in M phase. Here, we examined mitotic activation of endogenous c-Src, c-Yes, and Lyn, which are co-expressed in HeLa cells. c-Src, c-Yes, and Lyn were activated at different levels in M phase, and the activation was inhibited by Cdc2 inactivation. Mitotic c-Src and c-Yes exhibited normal- and retarded-electrophoretic-mobility forms on SDS-polyacrylamide gels, whereas Lyn did not show mobility retardation. Like c-Src, the retardation of electrophoretic mobility of c-Yes was caused by Cdc2-mediated phosphorylation. The normal- and retarded-mobility forms of c-Src were comparably activated, but activation of the retarded-mobility form of c-Yes was higher than that of the normal-mobility form of c-Yes. Thus, these results suggest that endogenous c-Src, c-Yes, and Lyn are differentially activated through Cdc2 activation during M phase.

  11. Src Kinase Inhibition Attenuates Morphine Tolerance without Affecting Reinforcement or Psychomotor Stimulation.

    Bull, Fiona A; Baptista-Hon, Daniel T; Sneddon, Claire; Wright, Lisa; Walwyn, Wendy; Hales, Tim G


    Prolonged opioid administration leads to tolerance characterized by reduced analgesic potency. Pain management is additionally compromised by the hedonic effects of opioids, the cause of their misuse. The multifunctional protein β-arrestin2 regulates the hedonic effects of morphine and participates in tolerance. These actions might reflect µ opioid receptor up-regulation through reduced endocytosis. β-Arrestin2 also recruits kinases to µ receptors. We explored the role of Src kinase in morphine analgesic tolerance, locomotor stimulation, and reinforcement in C57BL/6 mice. Analgesic (tail withdrawal latency; percentage of maximum possible effect, n = 8 to 16), locomotor (distance traveled, n = 7 to 8), and reinforcing (conditioned place preference, n = 7 to 8) effects of morphine were compared in wild-type, µ, µ, and β-arrestin2 mice. The influence of c-Src inhibitors dasatinib (n = 8) and PP2 (n = 12) was examined. Analgesia in morphine-treated wild-type mice exhibited tolerance, declining by day 10 to a median of 62% maximum possible effect (interquartile range, 29 to 92%). Tolerance was absent from mice receiving dasatinib. Tolerance was enhanced in µ mice (34% maximum possible effect; interquartile range, 5 to 52% on day 5); dasatinib attenuated tolerance (100% maximum possible effect; interquartile range, 68 to 100%), as did PP2 (91% maximum possible effect; interquartile range, 78 to 100%). By contrast, c-Src inhibition affected neither morphine-evoked locomotor stimulation nor reinforcement. Remarkably, dasatinib not only attenuated tolerance but also reversed established tolerance in µ mice. The ability of c-Src inhibitors to inhibit tolerance, thereby restoring analgesia, without altering the hedonic effect of morphine, makes c-Src inhibitors promising candidates as adjuncts to opioid analgesics.

  12. Targeting androgen receptor/Src complex impairs the aggressive phenotype of human fibrosarcoma cells.

    Castoria, Gabriella; Giovannelli, Pia; Di Donato, Marzia; Hayashi, Ryo; Arra, Claudio; Appella, Ettore; Auricchio, Ferdinando; Migliaccio, Antimo


    Hormones and growth factors influence the proliferation and invasiveness of human mesenchymal tumors. The highly aggressive human fibrosarcoma HT1080 cell line harbors classical androgen receptor (AR) that responds to androgens triggering cell migration in the absence of significant mitogenesis. As occurs in many human cancer cells, HT1080 cells also express epidermal growth factor receptor (EGFR). We report that the pure anti-androgen Casodex inhibits the growth of HT1080 cell xenografts in immune-depressed mice, revealing a novel role of AR in fibrosarcoma progression. In HT1080 cultured cells EGF, but not androgens, robustly increases DNA synthesis. Casodex abolishes the EGF mitogenic effect, implying a crosstalk between EGFR and AR. The mechanism underlying this crosstalk has been analyzed using an AR-derived small peptide, S1, which prevents AR/Src tyrosine kinase association and androgen-dependent Src activation. Present findings show that in HT1080 cells EGF induces AR/Src Association, and the S1 peptide abolishes both the assembly of this complex and Src activation. The S1 peptide inhibits EGF-stimulated DNA synthesis, cell matrix metalloproteinase-9 (MMP-9) secretion and invasiveness of HT1080 cells. Both Casodex and S1 peptide also prevent DNA synthesis and migration triggered by EGF in various human cancer-derived cells (prostate, breast, colon and pancreas) that express AR. This study shows that targeting the AR domain involved in AR/Src association impairs EGF signaling in human fibrosarcoma HT1080 cells. The EGF-elicited processes inhibited by the peptide (DNA synthesis, MMP-9 secretion and invasiveness) cooperate in increasing the aggressive phenotype of HT1080 cells. Therefore, AR represents a new potential therapeutic target in human fibrosarcoma, as supported by Casodex inhibition of HT1080 cell xenografts. The extension of these findings in various human cancer-derived cell lines highlights the conservation of this process across divergent cancer

  13. Caveolin-1 modulates cardiac gap junction homeostasis and arrhythmogenecity by regulating cSrc tyrosine kinase.

    Yang, Kai-Chien; Rutledge, Cody A; Mao, Mao; Bakhshi, Farnaz R; Xie, An; Liu, Hong; Bonini, Marcelo G; Patel, Hemal H; Minshall, Richard D; Dudley, Samuel C


    Genome-wide association studies have revealed significant association of caveolin-1 (Cav1) gene variants with increased risk of cardiac arrhythmias. Nevertheless, the mechanism for this linkage is unclear. Using adult Cav1(-/-) mice, we revealed a marked reduction in the left ventricular conduction velocity in the absence of myocardial Cav1, which is accompanied with increased inducibility of ventricular arrhythmias. Further studies demonstrated that loss of Cav1 leads to the activation of cSrc tyrosine kinase, resulting in the downregulation of connexin 43 and subsequent electric abnormalities. Pharmacological inhibition of cSrc mitigates connexin 43 downregulation, slowed conduction, and arrhythmia inducibility in Cav1(-/-) animals. Using a transgenic mouse model with cardiac-specific overexpression of angiotensin-converting enzyme (ACE8/8), we demonstrated that, on enhanced cardiac renin-angiotensin system activity, Cav1 dissociated from cSrc because of increased Cav1 S-nitrosation at Cys(156), leading to cSrc activation, connexin 43 reduction, impaired gap junction function, and subsequent increase in the propensity for ventricular arrhythmias and sudden cardiac death. Renin-angiotensin system-induced Cav1 S-nitrosation was associated with increased Cav1-endothelial nitric oxide synthase binding in response to increased mitochondrial reactive oxidative species generation. The present studies reveal the critical role of Cav1 in modulating cSrc activation, gap junction remodeling, and ventricular arrhythmias. These data provide a mechanistic explanation for the observed genetic link between Cav1 and cardiac arrhythmias in humans and suggest that targeted regulation of Cav1 may reduce arrhythmic risk in cardiac diseases associated with renin-angiotensin system activation. © 2014 American Heart Association, Inc.

  14. Activation of Src and release of intracellular calcium by phosphatidic acid during Xenopus laevis fertilization

    Bates, Ryan C.; Fees, Colby P.; Holland, William L.; Winger, Courtney C.; Batbayar, Khulan; Ancar, Rachel; Bergren, Todd; Petcoff, Douglas; Stith, Bradley J.


    We report a new step in the fertilization in Xenopus laevis which has been found to involve activation of Src tyrosine kinase to stimulate phospholipase C-γ (PLC- γ) which increases inositol 1,4,5-trisphosphate (IP3) to release intracellular calcium ([Ca]i). Molecular species analysis and mass measurements suggested that sperm activate phospholipase D (PLD) to elevate phosphatidic acid (PA). We now report that PA mass increased 2.7 fold by 1 minute after insemination and inhibition of PA production by two methods inhibited activation of Src and PLCγ, increased [Ca]i and other fertilization events. As compared to 14 other lipids, PA strongly bound Xenopus Src but not PLCγ. Addition of synthetic PA activated egg Src (an action requiring intact lipid rafts) and PLCγ as well as doubling the amount of PLCγ in rafts. In the absence of elevated [Ca]i, PA addition elevated IP3 mass to levels equivalent to that induced by sperm (but twice that achieved by calcium ionophore). Finally, PA induced [Ca]i release that was blocked by an IP3 receptor inhibitor. As only PLD1b message was detected, and Western blotting did not detect PLD2, we suggest that sperm activate PLD1b to elevate PA which then binds to and activates Src leading to PLCγ stimulation, IP3 elevation and [Ca]i release. Due to these and other studies, PA may also play a role in membrane fusion events such as sperm-egg fusion, cortical granule exocytosis, the elevation of phosphatidylinositol 4,5-bisphosphate and the large, late increase in sn 1,2-diacylglycerol in fertilization. PMID:24269904

  15. Provenance in Data Interoperability for Multi-Sensor Intercomparison

    Lynnes, C.; Leptoukh, G.; Berrick, S.; Shen, S.; Prados, A.; Fox, P.; Yang, W.; Min, M.; Holloway, D.; Enloe, Y.


    As our inventory of Earth science data sets grows, the ability to compare, merge and fuse multiple datasets grows in importance. This implies a need for deeper data interoperability than we have now. Many efforts (e.g. OPeNDAP, Open Geospatial Consortium) have broken down format barriers to interoperability; the next challenge is the semantic aspects of the data. Consider the issues when satellite data are merged, cross- calibrated, validated, inter-compared and fused. We must determine how to match up data sets that are related, yet different in significant ways: the exact nature of the phenomenon being measured, measurement technique, exact location in space-time, or the quality of the measurements. If subtle distinctions between similar measurements are not clear to the user, the results can be meaningless or even lead to an incorrect interpretation of the data. Most of these distinctions trace back to how the data came to be: sensors, processing, and quality assessment. For example, monthly averages of satellite-based aerosol measurements often show significant discrepancies, which might be due to differences in spatio-temporal aggregation, sampling issues, sensor biases, algorithm differences and/or calibration issues. This provenance information must therefore be captured in a semantic framework that allows sophisticated data inter-use tools to incorporate it, and eventually aid in the interpretation of comparison or merged products. Semantic web technology allows us to encode our knowledge of measurement characteristics, phenomena measured, space-time representations, and data quality representation in a well-structured, machine- readable ontology and rulesets. An analysis tool can use this knowledge to show users the provenance- related distinctions between two variables, advising on options for further data processing and analysis. An additional problem for workflows distributed across heterogeneous systems is retrieval and transport of provenance

  16. Mechanism for Src activation by the CCK2 receptor: Patho-physiological functions of this receptor in pancreas

    Audrey Ferrand; Sebastien Vatinel; Aline Kowalski-Chauvel; Claudine Bertrand; Chantal Escrieut; Daniel Fourmy; Marlene Dufresne; Catherine Seva


    AIM: To investigate in vivo, whether CCK2 receptors (CCK2R) regulate proteins known to play a crucial role in cell proliferation and cancer development and analyse in vitro the molecular mechanisms that lead to Src activation; in particular, to identify the domains within the CCK2R sequence that are implicated in this activation.METHODS: The expression and activation of Src and ERK were studied in vivo using immunofluorescence and western-blot techniques. We used pancreatic tissues derived from wild type or ElasCCK2 mice that expressed the CCK2R in pancreatic acini, displayed an increased pancreatic growth and developed preneoplastic lesions. The pancreatic tumor cell line AR4-2J expressing the endogenous CCK2R or COS-7 cells transiently transfected with wild type or mutant CCK2R were used as in vitro models to study the mechanism of Src activation.Src activation was measured by in vitro kinase assays, ERK activation by western blot using antiphospho-ERK antibodies and the involvement of Src in gastrin-induced cell proliferation by MTT test.RESULTS: We showed in vivo that the targeted CCK2R expression in the pancreas of Elas-CCK2 mice, led to the activation of Src and the ERK pathway. Src was activated upstream of the ERK pathway by the CCK2R in pancreatic tumoral cells and contributed to the proliferative effects mediated by this receptor. In vitro results demonstrated that activation of the Src/ERK pathway by the CCK2R required the NPXXY motif, located within the CCK2R sequence at the end of the 7th transmembrane domain,and suggested the putative role of Gq in this mechanism.CONCLUSION: Deregulation of the Src/ERK pathway by the CCK2R might represent an early step that contributes to cell proliferation, formation of preneoplastic lesions and pancreatic tumor development.

  17. Src tyrosine kinase regulates the stem cell factor–induced breakdown of the blood–retinal barrier

    Im, Ji-Eun; Song, Sun-Hwa


    Purpose Stem cell factor (SCF) has been recently acknowledged as a novel endothelial permeability factor. However, the mechanisms by which SCF-induced activation of the SCF cognate receptor, cKit, enhances endothelial permeability have not been fully elucidated. This study aimed to investigate the role of Src in SCF-induced breakdown of the blood–retinal barrier (BRB). Methods In vitro endothelial permeability and in vivo retinal vascular permeability assays were performed to investigate the role of Src in SCF-induced breakdown of the BRB. Immunofluorescence staining experiments were performed to analyze the cellular distribution of phosphorylated Src and vascular endothelial (VE)-cadherin. Results SCF markedly reduced electric resistance across the human retinal vascular endothelial monolayer in vitro and enhanced extravasation of dyes in murine retinal vasculature in vivo. Inhibition of cKit activation using cKit mutant mice and chemical inhibitor substantially diminished the ability of SCF to increase endothelial permeability and retinal vascular leakage. In human retinal vascular endothelial cells, SCF induced strong phosphorylation of Src and distinct localization of phosphorylated Src in the plasma membrane. Inhibition of Src activation using chemical inhibitors abolished the SCF-induced hyperpermeability of human retinal vascular endothelial cells and retinal vascular leakage in mice. In addition, treatment with Src inhibitors restored junctional expression of VE-cadherin that disappeared in SCF-treated retinal endothelial cells and retinal vasculature. Conclusions These results showed the important role of Src in mediating SCF-induced breakdown of the BRB and retinal vascular leakage. Given that increased retinal vascular permeability is a common manifestation of various ocular diseases, the SCF/cKit/Src signaling pathway may be involved in the development of the hyperpermeable retinal vasculature in many ocular disorders.

  18. SRC-1 quarterly technical report, April-June 1981


    Volume 2 consists of further evaluations required for the design of the demonstration plant: in particular, (1) a comparison of the performance and analytical characterization of several recycle solvents which developed in various pilot plants or with specific coals, and (2) the catalytic effects of residues or minerals on dissolver performance (There are several papers on the latter problem; the results are relatively small but significant in terms of plant performance and somewhat complex; it may help to read the last paper first, since the nature of the results was somewhat better understood by the time it was written and some problems and inconsistenices resolved to some degree). (LTN)

  19. Improved response by co-targeting EGFR/EGFRvIII and Src family kinases in human cancer cells

    Andersen, Peter; Villingshøj, Mette; Poulsen, Hans Skovgaard


    We hypothesized that co-targeting the epidermal growth factor receptor (EGFR) and Src with the EGFR inhibitor gefitinib and the Src inhibitor AZD0530 would increase growth inhibition and impede migration. Cells overexpressing EGFR were more sensitive to gefitinib than cells expressing mutated EGFR...... or normal levels of wild-type EGFR. Furthermore, cells with mutated EGFR responded to low doses of gefitinib with increased proliferation. AZD0530 was an effective inhibitor of proliferation and migration, irrespective of EGFR status. These results suggest that co-targeting EGFR and Src might be a valuable...

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

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


    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.

  1. Improved response by co-targeting EGFR/EGFRvIII and Src family kinases in human cancer cells

    Andersen, Peter; Villingshøj, Mette; Poulsen, Hans Skovgaard


    or normal levels of wild-type EGFR. Furthermore, cells with mutated EGFR responded to low doses of gefitinib with increased proliferation. AZD0530 was an effective inhibitor of proliferation and migration, irrespective of EGFR status. These results suggest that co-targeting EGFR and Src might be a valuable......We hypothesized that co-targeting the epidermal growth factor receptor (EGFR) and Src with the EGFR inhibitor gefitinib and the Src inhibitor AZD0530 would increase growth inhibition and impede migration. Cells overexpressing EGFR were more sensitive to gefitinib than cells expressing mutated EGFR...

  2. Provenance-based refresh in data-oriented workflows

    Ikeda, Robert


    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.

  3. Sudetic larch in Germany - Results of provenance and progeny research

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


    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

  4. Provenance Usage in the OceanLink Project

    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.


    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.

  5. The Src family tyrosine kinases src and yes have differential effects on inflammation-induced apoptosis in human pulmonary microvascular endothelial cells.

    Nelin, Leif D; White, Hilary A; Jin, Yi; Trittmann, Jennifer K; Chen, Bernadette; Liu, Yusen


    Endothelial cells are essential for normal lung function: they sense and respond to circulating factors and hemodynamic alterations. In inflammatory lung diseases such as acute respiratory distress syndrome, endothelial cell apoptosis is an inciting event in pathogenesis and a prominent pathological feature. Endothelial cell apoptosis is mediated by circulating inflammatory factors, which bind to receptors on the cell surface, activating signal transduction pathways, leading to caspase-3-mediated apoptosis. We hypothesized that yes and src have differential effects on caspase-3 activation in human pulmonary microvascular endothelial cells (hPMVEC) due to differential downstream signaling effects. To test this hypothesis, hPMVEC were treated with siRNA against src (siRNAsrc), siRNA against yes (siRNAyes), or their respective scramble controls. After recovery, the hPMVEC were treated with cytomix (LPS, IL-1β, TNF-α, and IFN-γ). Treatment with cytomix induced activation of the extracellular signal-regulated kinase (ERK) pathway and caspase-3-mediated apoptosis. Treatment with siRNAsrc blunted cytomix-induced ERK activation and enhanced cleaved caspase-3 levels, while treatment with siRNAyes enhanced cytomix-induced ERK activation and attenuated levels of cleaved caspase-3. Inhibition of the ERK pathway using U0126 enhanced cytomix-induced caspase-3 activity. Treatment of hPMVEC with cytomix induced Akt activation, which was inhibited by siRNAsrc. Inhibition of the phosphatidylinositol 3-kinase/Akt pathway using LY294002 prevented cytomix-induced ERK activation and augmented cytomix-induced caspase-3 cleavage. Together, our data demonstrate that, in hPMVEC, yes activation blunts the ERK cascade in response to cytomix, resulting in greater apoptosis, while cytomix-induced src activation induces the phosphatidylinositol 3-kinase pathway, which leads to activation of Akt and ERK and attenuation of apoptosis.

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

    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: [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: [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)


    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.

  7. 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.

    Chellaiah, Meenakshi A; Schaller, Michael D


    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.

  8. A Provenance-based Trust Model for Delay Tolerant Networks


    data derivation, and data annotation [10]. Since then, OPM has been widely adopted and extended by various research groups [8]. Freire et al. [5...VLDB Workshop on Secure Data Management, LNCS, vol. 5159, pp. 82–98, Auckland, New Zealand, Aug. 24, 2008. 5. J. Freire , D. Koop, E. Santos, and C.T... Freire , J. Futrelle, R.E. McGrath, J. Myers, and P. Paulson, “The open provenance model: an overview,” Int’l Provenance and Annotation Workshop, Salt Lake

  9. Structural insights into the recognition of β3 integrin cytoplasmic tail by the SH3 domain of Src kinase.

    Katyal, Priya; Puthenveetil, Robbins; Vinogradova, Olga


    Src kinase plays an important role in integrin signaling by regulating cytoskeletal organization and cell remodeling. Previous in vivo studies have revealed that the SH3 domain of c-Src kinase directly associates with the C-terminus of β3 integrin cytoplasmic tail. Here, we explore this binding interface with a combination of different spectroscopic and computational methods. Chemical shift mapping, PRE, transferred NOE and CD data were used to obtain a docked model of the complex. This model suggests a different binding mode from the one proposed through previous studies wherein, the C-terminal end of β3 spans the region in between the RT and n-Src loops of SH3 domain. Furthermore, we show that tyrosine phosphorylation of β3 prevents this interaction, supporting the notion of a constitutive interaction between β3 integrin and Src kinase.

  10. Separating stimulus-driven and response-related LRP components with Residue Iteration Decomposition (RIDE).

    Stürmer, Birgit; Ouyang, Guang; Zhou, Changsong; Boldt, Annika; Sommer, Werner


    When the lateralized readiness potential (LRP) is recorded in stimulus-response compatibility (SRC) tasks, two processes may overlap in the LRP, stimulus-driven response priming and activation based on response selection rules. These overlapping processes are hard to disentangle with standard analytical tools. Here, we show that Residue Iteration Decomposition (RIDE), based on latency variability, separates the overlapping LRP components from a Simon task into stimulus-driven and response-related components. SRC affected LRP amplitudes only in the stimulus-driven component, whereas LRP onsets were affected only in the response-locked component. Importantly, the compatibility effect in reaction times was more similar to the effect in the onsets of the RIDE-derived response-locked LRP component than in the unseparated LRP. Thus, RIDE-separated LRP components are devoid of distortions inherent to standard LRPs.

  11. Src-mediated phosphorylation of the tyrosine phosphatase PRL-3 is required for PRL-3 promotion of Rho activation, motility and invasion.

    Fiordalisi, James J; Dewar, Brian J; Graves, Lee M; Madigan, James P; Cox, Adrienne D


    The metastasis-associated tyrosine phosphatase PRL-3/PTP4A is upregulated in numerous cancers, but the mechanisms modulating PRL-3 activity other than its expression levels have not been investigated. Here we report evidence for both Src-dependent tyrosine phosphorylation of PRL-3 and Src-mediated regulation of PRL-3 biological activities. We used structural mutants, pharmacological inhibitors and siRNA to demonstrate Src-dependent phosphorylation of endogenous PRL-3 in SW480 colon cancer cells. We also demonstrated that PRL-3 was not tyrosine phosphorylated in SYF mouse embryo fibroblasts deficient in Src, Yes and Fyn unless Src was re-expressed. Further, we show that platelet-derived growth factor (PDGF) can stimulate PRL-3 phosphorylation in a Src-dependent manner. Finally, we show that PRL-3-induced cell motility, Matrigel invasion and activation of the cytoskeleton-regulating small GTPase RhoC were abrogated in the presence of the phosphodeficient PRL-3 mutant Y53F, or by use of a Src inhibitor. Thus, PRL-3 requires the activity of a Src kinase, likely Src itself, to promote these cancer-associated phenotypes. Our data establish a model for the regulation of PRL-3 by Src that supports the possibility of their coordinate roles in signaling pathways promoting invasion and metastasis, and supports simultaneous use of novel molecularly targeted therapeutics directed at these proteins.

  12. Sepsis-induced cardiac mitochondrial dysfunction involves altered mitochondrial-localization of tyrosine kinase Src and tyrosine phosphatase SHP2.

    Qun S Zang

    Full Text Available Our previous research demonstrated that sepsis produces mitochondrial dysfunction with increased mitochondrial oxidative stress in the heart. The present study investigated the role of mitochondria-localized signaling molecules, tyrosine kinase Src and tyrosine phosphatase SHP2, in sepsis-induced cardiac mitochondrial dysfunction using a rat pneumonia-related sepsis model. SD rats were given an intratracheal injection of Streptococcus pneumoniae, 4×10(6 CFU per rat, (or vehicle for shams; heart tissues were then harvested and subcellular fractions were prepared. By Western blot, we detected a gradual and significant decrease in Src and an increase in SHP2 in cardiac mitochondria within 24 hours post-inoculation. Furthermore, at 24 hours post-inoculation, sepsis caused a near 70% reduction in tyrosine phosphorylation of all cardiac mitochondrial proteins. Decreased tyrosine phosphorylation of certain mitochondrial structural proteins (porin, cyclophilin D and cytochrome C and functional proteins (complex II subunit 30kD and complex I subunit NDUFB8 were evident in the hearts of septic rats. In vitro, pre-treatment of mitochondrial fractions with recombinant active Src kinase elevated OXPHOS complex I and II-III activity, whereas the effect of SHP2 phosphatase was opposite. Neither Src nor SHP2 affected complex IV and V activity under the same conditions. By immunoprecipitation, we showed that Src and SHP2 consistently interacted with complex I and III in the heart, suggesting that complex I and III contain putative substrates of Src and SHP2. In addition, in vitro treatment of mitochondrial fractions with active Src suppressed sepsis-associated mtROS production and protected aconitase activity, an indirect marker of mitochondrial oxidative stress. On the contrary, active SHP2 phosphatase overproduced mtROS and deactivated aconitase under the same in vitro conditions. In conclusion, our data suggest that changes in mitochondria

  13. Global tyrosine kinome profiling of human thyroid tumors identifies Src as a promising target for invasive cancers

    Cho, Nancy L., E-mail: [Department of Surgery, Brigham and Women' s Hospital, Boston, MA 02115 (United States); Lin, Chi-Iou [Department of Surgery, Brigham and Women' s Hospital, Boston, MA 02115 (United States); Du, Jinyan [Broad Institute, Massachusetts Institute of Technology, Cambridge, MA 02142 (United States); Whang, Edward E. [Department of Surgery, Brigham and Women' s Hospital, Boston, MA 02115 (United States); Ito, Hiromichi [Department of Surgery, Michigan State University, Lansing, MI 48912 (United States); Moore, Francis D.; Ruan, Daniel T. [Department of Surgery, Brigham and Women' s Hospital, Boston, MA 02115 (United States)


    Highlights: Black-Right-Pointing-Pointer Kinome profiling is a novel technique for identifying activated kinases in human cancers. Black-Right-Pointing-Pointer Src activity is increased in invasive thyroid cancers. Black-Right-Pointing-Pointer Inhibition of Src activity decreased proliferation and invasion in vitro. Black-Right-Pointing-Pointer Further investigation of Src targeted therapies in thyroid cancer is warranted. -- Abstract: Background: Novel therapies are needed for the treatment of invasive thyroid cancers. Aberrant activation of tyrosine kinases plays an important role in thyroid oncogenesis. Because current targeted therapies are biased toward a small subset of tyrosine kinases, we conducted a study to reveal novel therapeutic targets for thyroid cancer using a bead-based, high-throughput system. Methods: Thyroid tumors and matched normal tissues were harvested from twenty-six patients in the operating room. Protein lysates were analyzed using the Luminex immunosandwich, a bead-based kinase phosphorylation assay. Data was analyzed using GenePattern 3.0 software and clustered according to histology, demographic factors, and tumor status regarding capsular invasion, size, lymphovascular invasion, and extrathyroidal extension. Survival and invasion assays were performed to determine the effect of Src inhibition in papillary thyroid cancer (PTC) cells. Results: Tyrosine kinome profiling demonstrated upregulation of nine tyrosine kinases in tumors relative to matched normal thyroid tissue: EGFR, PTK6, BTK, HCK, ABL1, TNK1, GRB2, ERK, and SRC. Supervised clustering of well-differentiated tumors by histology, gender, age, or size did not reveal significant differences in tyrosine kinase activity. However, supervised clustering by the presence of invasive disease showed increased Src activity in invasive tumors relative to non-invasive tumors (60% v. 0%, p < 0.05). In vitro, we found that Src inhibition in PTC cells decreased cell invasion and proliferation

  14. Ca2+/Calmodulin and Apo-Calmodulin Both Bind to and Enhance the Tyrosine Kinase Activity of c-Src.

    Silviya R Stateva

    Full Text Available Src family non-receptor tyrosine kinases play a prominent role in multiple cellular processes, including: cell proliferation, differentiation, cell survival, stress response, and cell adhesion and migration, among others. And when deregulated by mutations, overexpression, and/or the arrival of faulty incoming signals, its hyperactivity contributes to the development of hematological and solid tumors. c-Src is a prototypical member of this family of kinases, which is highly regulated by a set of phosphorylation events. Other factor contributing to the regulation of Src activity appears to be mediated by the Ca2+ signal generated in cells by different effectors, where the Ca2+-receptor protein calmodulin (CaM plays a key role. In this report we demonstrate that CaM directly interacts with Src in both Ca2+-dependent and Ca2+-independent manners in vitro and in living cells, and that the CaM antagonist N-(6-aminohexyl-5-chloro-1-naphthalenesulfonamide (W-7 inhibits the activation of this kinase induced by the upstream activation of the epidermal growth factor receptor (EGFR, in human carcinoma epidermoide A431 cells, and by hydrogen peroxide-induced oxidative stress, in both A431 cells and human breast adenocarcinoma SK-BR-3 cells. Furthermore, we show that the Ca2+/CaM complex strongly activates the auto-phosphorylation and tyrosine kinase activity of c-Src toward exogenous substrates, but most relevantly and for the first time, we demonstrate that Ca2+-free CaM (apo-CaM exerts a far higher activatory action on Src auto-phosphorylation and kinase activity toward exogenous substrates than the one exerted by the Ca2+/CaM complex. This suggests that a transient increase in the cytosolic concentration of free Ca2+ is not an absolute requirement for CaM-mediated activation of Src in living cells, and that a direct regulation of Src by apo-CaM could be inferred.

  15. c-Yes tyrosine kinase is a potent suppressor of ES cell differentiation and antagonizes the actions of its closest phylogenetic relative, c-Src.

    Zhang, Xiong; Meyn, Malcolm A; Smithgall, Thomas E


    Embryonic stem (ES) cells are derived from the inner cell mass of the blastocyst stage embryo and are characterized by self-renewal and pluripotency. Previous work has shown that Src-family tyrosine kinases display dynamic expression and activity changes during ES cell differentiation, suggesting distinct functions in the control of developmental fate. Here we used ES cells to test the hypothesis that c-Src and its closest phylogenetic relative, c-Yes, act in biological opposition despite their strong homology. Unlike c-Src, enforced expression of active c-Yes blocked ES cell differentiation to embryoid bodies by maintaining pluripotency gene expression. To explore the interplay of c-Src and c-Yes in ES cell differentiation, we engineered c-Src and c-Yes mutants that are resistant to A-419259, a potent pyrrolopyrimidine inhibitor of the Src kinase family. Previous studies have shown that A-419259 treatment blocks all Src-family kinase activity in ES cells, preventing differentiation while maintaining pluripotency. Expression of inhibitor-resistant c-Src but not c-Yes rescued the A-419259 differentiation block, resulting in a cell population with properties of both primitive ectoderm and endoderm. Remarkably, when inhibitor-resistant c-Src and c-Yes were expressed together in ES cells, c-Yes activity suppressed c-Src-mediated differentiation. These studies show that even closely related kinases such as c-Src and c-Yes have unique and opposing functions in the same cell type. Selective agonists or inhibitors of c-Src versus c-Yes activity may allow more precise pharmacological manipulation of ES cell fate and have broader applications in other biological systems that express multiple Src family members such as tumor cells.

  16. Modulation of FAK and Src adhesion signaling occurs independently of adhesion complex composition

    Horton, Edward R.; Humphries, Jonathan D.; Stutchbury, Ben


    of the key IAC signaling components focal adhesion kinase (FAK) and Src. FAK inhibition using AZ13256675 blocked FAKY397 phosphorylation but did not alter IAC composition, as reported by mass spectrometry. IAC composition was also insensitive to Src inhibition using AZD0530 alone or in combination with FAK...... inhibition. In contrast, kinase inhibition substantially reduced phosphorylation within IACs, cell migration and proliferation. Furthermore using fluorescence recovery after photobleaching, we found that FAK inhibition increased the exchange rate of a phosphotyrosine (pY) reporter (dSH2) at IACs. These data...... demonstrate that kinase-dependent signal propagation through IACs is independent of gross changes in IAC composition. Together, these findings demonstrate a general separation between the composition of IACs and their ability to relay pY-dependent signals....

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

    Sherman, Alexander Charles


    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.

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

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


    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.

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


    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.

  20. The release of glutamate from cortical neurons regulated by BDNF via the TrkB/Src/PLC-γ1 pathway.

    Zhang, Zitao; Fan, Jin; Ren, Yongxin; Zhou, Wei; Yin, Guoyong


    The brain-derived neurotrophic factor (BDNF) participates in the regulation of cortical neurons by influencing the release of glutamate. However, the specific mechanisms are unclear. Hence, we isolated and cultured the cortical neurons of Sprague Dawley rats. Specific inhibitors of TrkB, Src, PLC-γ1, Akt, and MEK1/2 (i.e., K252a, PP2, U73122, LY294002, and PD98059, respectively) were used to treat cortical neurons and to detect the glutamate release from cortical neurons stimulated with BDNF. BDNF significantly increased glutamate release, and simultaneously enhanced phosphorylation levels of TrkB, Src, PLC-γ, Akt, and Erk1/2. For BDNF-stimulated cortical neurons, K252a inhibited glutamate release and inhibited the phosphorylation levels of TrkB, Src, PLC-γ, Erk1/2, and Akt (P PLC-γ1 (P 0.05). U73122 inhibited the glutamate release from BDNF-stimulated cortical neurons, but had no influence on the phosphorylation levels of TrkB, Src, Erk1/2, or Akt (P > 0.05). LY294002 and PD98059 did not affect the BDNF-stimulated glutamate release and did not inhibit the phosphorylation levels of TrkB, Src, or PLC-γ1. In summary, BDNF stimulated the glutamate release from cortical neurons via the TrkB/Src/PLC-γ1 signaling pathway.

  1. Facilitating Fine Grained Data Provenance using Temporal Data Model

    Huq, M.R.; Wombacher, Andreas; Apers, Peter M.G.


    E-science applications use fine grained data provenance to maintain the reproducibility of scientific results, i.e., for each processed data tuple, the source data used to process the tuple as well as the used approach is documented. Since most of the e-science applications perform on-line

  2. Trace elements in quartz shed light on sediment provenance

    Ackerson, Michael R.; Tailby, Nicholas D.; Watson, E. Bruce


    Quartz is one of the most common minerals on the surface of the earth, and is a primary rock-forming mineral across the rock cycle. These two factors make quartz an obvious target for sediment provenance studies. Observations from experimental and natural samples demonstrate that the trace element content of quartz often reflects the conditions of quartz formation. When quartz is weathered from its primary crystallization setting (i.e., quartz from a granitoid) it can retain many chemical signatures of formation throughout the sedimentation processes. These geochemical signatures can be used to understand the primary source of individual quartz grains within a sediment. Here we present a case study from the Bega River catchment to demonstrate that quartz grains in sediments at the mouth of the Bega River are sourced from granitoids within the drainage basin. Data presented here also indicate that a portion of the beach sediment is also derived from either (i) sedimentary rocks within the basin or; (ii) mixing with sediments at the mouth of the river. The Bega River catchment was selected for this study because it is both small and has a well-constrained bedrock lithology, making it an ideal location to test the utility of this provenance technique. However, quartz trace element provenance has broad applications to modern and ancient sediments and can be used in lieu of, or in conjunction with, other provenance techniques to elucidate sediment transport through time.

  3. Seasonal growth in white pine seedlings from different provenances

    Frank S., Jr. Santamour


    The Northeastern Forest Experiment Station, in cooperation with other experiment stations in the United States and Canada, began a range-wide provenance test of eastern white pine (Pinus strobus L.) in 1955. Seed was collected from 31 different locations in 17 states and 4 Canadian provinces. In most places collections were made from 10 trees at each location. The seed...

  4. Probabilistic inference of fine-grained data provenance

    Huq, Mohammad Rezwanul; Apers, P.M.G.; Wombacher, A.


    Decision making, process control and e-science applications process stream data, mostly produced by sensors. To control and monitor these applications, reproducibility of result is a vital requirement. However, it requires massive amount of storage space to store fine-grained provenance data especia

  5. Position Paper: Provenance Data Visualisation for Neuroimaging Analysis

    Arshad, B.; Munir, K.; McClatchey, R.; Liaquat Kiani, S.


    Visualisation facilitates the understanding of scientific data both through exploration and explanation of visualised data. Provenance contributes to the understanding of data by containing the contributing factors behind a result. With the significant increase in data volumes and algorithm complexity, clinical researchers are struggling with information tracking, analysis reproducibility and the verification of scientific output. Data coming from various heterogeneous sources (multiple sourc...

  6. Using provenance to manage knowledge of in silico experiments.

    Stevens, Robert; Zhao, Jun; Goble, Carole


    This article offers a briefing in one of the knowledge management issues of in silico experimentation in bioinformatics. Recording of the provenance of an experiment-what was done; where, how and why, etc. is an important aspect of scientific best practice that should be extended to in silico experimentation. We will do this in the context of eScience which has been part of the move of bioinformatics towards an industrial setting. Despite the computational nature of bioinformatics, these analyses are scientific and thus necessitate their own versions of typical scientific rigour. Just as recording who, what, why, when, where and how of an experiment is central to the scientific process in laboratory science, so it should be in silico science. The generation and recording of these aspects, or provenance, of an experiment are necessary knowledge management goals if we are to introduce scientific rigour into routine bioinformatics. In Silico experimental protocols should themselves be a form of managing the knowledge of how to perform bioinformatics analyses. Several systems now exist that offer support for the generation and collection of provenance information about how a particular in silico experiment was run, what results were generated, how they were generated, etc. In reviewing provenance support, we will review one of the important knowledge management issues in bioinformatics.

  7. Provenances and fertilizer on early growth cedar seedlings

    Marcio Carlos Navroski


    Full Text Available The aim of the study was to evaluate the initial development of different provenances and the influence of base fertilizer and coverage on growth of Cedrela fissilis seedlings. Provenances of seeds were collected in Lapa, PR, Fernandes Pinheiro, PR and Itaara, RS. After germination, the seedlings were transplanted to plastic bags of 500 cm³, filled with commercial substrate. Total height (h, stem diameter (sd, and ratio h/sd seedlings were measured after 150 days of transplanting. Seedlings of Fernandes Pinheiro received basic fertilization after transplantation (0, 2, 4, 6 and 8 g dm-3  Osmocote® and cover (3 and 6 g L-1, respectively, of Peter’s® and urea. The provenance and doses of controlled-release fertilizer influenced early development of Cedrela fissilis seedlings. Itaara provenance showed better seedlings growth. Cedar seedlings showed good growth when incorporated into the substrate 5 g dm-3 Osmocote® and, in addition, applied in topdressing 3 g L-1 of Peter’s®. Urea topdressing is rarely recommended for cedar seedlings.

  8. Coactivator SRC-2–dependent metabolic reprogramming mediates prostate cancer survival and metastasis

    Dasgupta, Subhamoy; Putluri, Nagireddy; Long, Weiwen; Zhang, Bin; Wang, Jianghua; Kaushik, Akash K.; Arnold, James M.; Bhowmik, Salil K.; Stashi, Erin; Brennan, Christine A; Rajapakshe, Kimal; Coarfa, Cristian; Mitsiades, Nicholas; Ittmann, Michael M.; Chinnaiyan, Arul M


    Metabolic pathway reprogramming is a hallmark of cancer cell growth and survival and supports the anabolic and energetic demands of these rapidly dividing cells. The underlying regulators of the tumor metabolic program are not completely understood; however, these factors have potential as cancer therapy targets. Here, we determined that upregulation of the oncogenic transcriptional coregulator steroid receptor coactivator 2 (SRC-2), also known as NCOA2, drives glutamine-dependent de novo lip...

  9. Mutual Regulation of Src Family Kinases and the Neurotrophin Receptor TrkB*

    Huang, Yang Z.; McNamara, James O.


    The neurotrophin receptor tyrosine kinase TrkB is critical to diverse biological processes. We investigated the interplay of Src family kinases (SFKs) and TrkB to better understand mechanisms of TrkB signaling in physiological and pathological conditions. We compared and contrasted the role of SFKs in TrkB signaling following activation of TrkB by two mechanisms, its transactivation by zinc, and its activation by its prototypic neurotrophin ligand, brain-derived neurotrophic factor (BDNF). Us...

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


    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.

  11. Kinetics characterization of c-Src binding to lipid membranes: Switching from labile to persistent binding.

    Le Roux, Anabel-Lise; Busquets, Maria Antònia; Sagués, Francesc; Pons, Miquel


    Cell signaling by the c-Src proto-oncogen requires the attachment of the protein to the inner side of the plasma membrane through the myristoylated N-terminal region, known as the SH4 domain. Additional binding regions of lower affinity are located in the neighbor intrinsically disordered Unique domain and the structured SH3 domain. Here we present a surface plasmon resonance study of the binding of a myristoylated protein including the SH4, Unique and SH3 domains of c-Src to immobilized liposomes. Two distinct binding processes were observed: a fast and a slow one. The second process lead to a persistently bound form (PB) with a slower binding and a much slower dissociation rate than the first one. The association and dissociation of the PB form could be detected using an anti-SH4 antibody. The kinetic analysis revealed that binding of the PB form follows a second order rate law suggesting that it involves the formation of c-Src dimers on the membrane surface. A kinetically equivalent PB form is observed in a myristoylated peptide containing only the SH4 domain but not in a construct including the three domains but with a 12-carbon lauroyl substituent instead of the 14-carbon myristoyl group. The PB form is observed with neutral lipids but its population increases when the immobilized liposomes contain negatively charged lipids. We suggest that the PB form may represent the active signaling form of c-Src while the labile form provides the capacity for fast 2D search of the target signaling site on the membrane surface.

  12. Dynamin Forms a Src Kinase–sensitive Complex with Cbl and Regulates Podosomes and Osteoclast Activity


    Podosomes are highly dynamic actin-containing adhesion structures found in osteoclasts, macrophages, and Rous sarcoma virus (RSV)-transformed fibroblasts. After integrin engagement, Pyk2 recruits Src and the adaptor protein Cbl, forming a molecular signaling complex that is critical for cell migration, and deletion of any molecule in this complex disrupts podosome ring formation and/or decreases osteoclast migration. Dynamin, a GTPase essential for endocytosis, is also involved in actin cytos...

  13. Dynamin Reduces Pyk2 Y402 Phosphorylation and Src Binding in Osteoclasts ▿ †


    Signaling via the Pyk2-Src-Cbl complex downstream of integrins contributes to the assembly, organization, and dynamics of podosomes, which are the transient adhesion complexes of highly motile cells such as osteoclasts and dendritic cells. We previously demonstrated that the GTPase dynamin is associated with podosomes, regulates actin flux in podosomes, and promotes bone resorption by osteoclasts. We report here that dynamin associates with Pyk2, independent of dynamin's GTPase activity, and ...

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

    Siddiqui Ruqaiyyah; Iqbal Junaid; Maugueret Marie-josée; Khan Naveed


    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...

  15. Early redox, Src family kinase, and calcium signaling integrate wound responses and tissue regeneration in zebrafish

    Yoo, Sa Kan; Freisinger, Christina M.; LeBert, Danny C.; Huttenlocher, Anna


    Tissue injury can lead to scar formation or tissue regeneration. How regenerative animals sense initial tissue injury and transform wound signals into regenerative growth is an unresolved question. Previously, we found that the Src family kinase (SFK) Lyn functions as a redox sensor in leukocytes that detects H2O2 at wounds in zebrafish larvae. In this paper, using zebrafish larval tail fins as a model, we find that wounding rapidly activated SFK and calcium signaling in epithelia. The immedi...

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

    Shaoying Lu

    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.

  17. mGluR1-mediated excitation of cerebellar GABAergic interneurons requires both G protein-dependent and Src-ERK1/2-dependent signaling pathways.

    Hideo Kubota

    Full Text Available Stimulation of type I metabotropic glutamate receptors (mGluR1/5 in several neuronal types induces slow excitatory responses through activation of transient receptor potential canonical (TRPC channels. GABAergic cerebellar molecular layer interneurons (MLIs modulate firing patterns of Purkinje cells (PCs, which play a key role in cerebellar information processing. MLIs express mGluR1, and activation of mGluR1 induces an inward current, but its precise intracellular signaling pathways are unknown. We found that mGluR1 activation facilitated spontaneous firing of mouse cerebellar MLIs through an inward current mediated by TRPC1 channels. This mGluR1-mediated inward current depends on both G protein-dependent and -independent pathways. The nonselective protein tyrosine kinase inhibitors genistein and AG490 as well as the selective extracellular signal-regulated kinase 1/2 (ERK1/2 inhibitors PD98059 and SL327 suppressed the mGluR1-mediated current responses. Following G protein blockade, the residual mGluR1-mediated inward current was significantly reduced by the selective Src tyrosine kinase inhibitor PP2. In contrast to cerebellar PCs, GABAB receptor activation in MLIs did not alter the mGluR1-mediated inward current, suggesting that there is no cross-talk between mGluR1 and GABAB receptors in MLIs. Thus, activation of mGluR1 facilitates firing of MLIs through the TRPC1-mediated inward current, which depends on not only G protein-dependent but also Src-ERK1/2-dependent signaling pathways, and consequently depresses the excitability of cerebellar PCs.

  18. Protein tyrosine phosphatase PTPRB regulates Src phosphorylation and tumour progression in NSCLC.

    Qi, Yinliang; Dai, Yuanchang; Gui, Shuyu


    Protein tyrosine-phosphatases (PTPs) play important roles in various biological processes. Deregulation in PTP function has been implicated in carcinogenesis and tumour progression in many cancer types. However, the role of protein tyrosine phosphatase receptor type B (PTPRB) in non-small-cell lung cancer (NSCLC) tumorigenesis has not been investigated. Lentiviral vector expressing PTPRB cDNA or shRNA was infected into A549 and H1299 cell lines, followed by cell proliferation, colony formation, soft agar and invasion assays. A549 xenograft mouse model was used to evaluate in vivo function of PTPRB. Quantitative polymerase chain reaction (PCR) was used to measure PTPRB expression in NSCLC patient samples. Kaplan Meier analysis was performed to assess association between PTPRB expression and patient overall survival (OS). Multivariate analysis was performed to evaluate prognostic significance of PTPRB. Overexpression of PTPRB reduced cell proliferation rate, colony formation efficiency, soft agar growth and cell invasion in A549 and H1299 cells, as well as tumour growth rate in A549 xenograft. Knockdown of PTPRB increased Src phosphorylation and cell invasion, which was reversed by Src inhibitor PP2. Additionally, PTPRB was down-regulated in NSCLC patient and was associated with patient OS. PTPRB regulates Src phosphorylation and tumorigenesis in NSCLC. PTPRB may serve as an independent prognostic biomarker for NSCLC patients.

  19. Treatment of Recurrent Intracranial Hemangiopericytoma with SRC-Related Tyrosine Kinase Targeted Therapy: A Case Report

    Katherine B. Peters


    Full Text Available Hemangiopericytoma (HPC is a rare sarcomatous tumor arising from pericytes, a support cell found in blood vessels. These tumors can occur throughout the body, particularly in the lower extremities and retroperitoneum. In rare circumstances, HPCs can arise from the meninges. In these cases, they behave similar to meningiomas, in particular angiomatous meningiomas, but tend to be more aggressive and are likely to recur. Treatment usually focuses on surgical resection and radiotherapy with possible inclusion of chemotherapy for control of recurrent disease. We describe a case of recurrent right temporal HPC that first manifested as a paraneoplastic syndrome of oncogenic osteomalacia. Despite maximum therapy, this patient experienced multiple recurrences of the tumor, and immunohistochemical analysis revealed overexpression of platelet-derived growth factor receptor, a member of the SRC-related tyrosine kinases. After multiple recurrences, the patient’s tumor has been stable with treatment with monotherapy utilizing molecularly targeted therapy to SRC-related tyrosine kinases. This is the first case report of the treatment of recurrent meningeal HPC with molecularly targeted therapy to SRC-related tyrosine kinases.

  20. MiR-31 regulates the cisplatin resistance by targeting Src in gallbladder cancer.

    Li, Maolan; Chen, Wei; Zhang, Hongchen; Zhang, Yong; Ke, Fayong; Wu, Xiangsong; Zhang, Yijian; Weng, Mingzhe; Liu, Yingbin; Gong, Wei


    Gallbladder cancer (GBC) is a malignant tumor highly resistant to chemotherapy. MicroRNAs (miRNAs) are found extensively involved in modulation of carcinogenesis and chemoresistance. This study aimed to investigate cisplatin (DDP)-susceptibility regulated by expression of the miRNAs and underlying pathways in GBC. The microRNA-31 (miR-31) was selected by microarray due to the biggest fold change between DDP-resistant and parental cells. Ectopic overexpression of miR-31 decreased cell proliferation, viability and invasion capacity, but promoted apoptosis in DDP-resistant cells and in xenograft tumor models. Cell apoptosis and DDP-chemosensitivity was remarkably increased by knockdown of Src proto-oncogene (Src) expression, which was subsequently reversed by rescue of Src expression in miR-31-expressing cells. The microarray was used to select the candidate miRNA in two DDP-resistant GBC cell lines. The effect of regulated expression of the miRNA on cell migration, invasion, proliferation and apoptosis was examined by wound healing, transwell assays, CCK-8 assays, colony formation and flow cytometry assays, respectively. Xenograft tumor models were used to validate the function of the downstream target. Our results demonstrated that miR-31reduced significantly in GBC cells rendering resistance to cisplatin, and upregulated expression of miR-31 augmented chemosensitivity, presenting a therapeutic potential to overcome drug resistance in GBC.

  1. C-src enriched serum microvesicles are generated in malignant plasma cell dyscrasia.

    Giuseppe Di Noto

    Full Text Available Plasma cell dyscrasias are immunosecretory disorders that can lead to hematological malignancies such as Multiple Myeloma (MM. MM accounts for 15% of all hematologic cancers, and those diagnosed with MM typically become severely ill and have a low life expectancy. Monoclonal immunoglobulin Free Light Chains (FLC are present in the serum and urine of many patients with plasma cell diseases. The biological differences between monoclonal FLCs, produced under malignant or benign dyscrasias, has not yet been characterized. In the present study, we show that endothelial and heart muscle cell lines internalize kappa and lambda FLCs. After internalization, FLCs are rerouted in the extracellular space via microvesicles and exosomes that can be re-internalized in contiguous cells. Only FLCs secreted from malignant B Lymphocytes were carried in Hsp70, annexin V, and c-src positive vesicles. In both MM and AL Amyloidosis patients we observed an increase in microvesicle and exosome production. Isolated serum vesicles from MM, AL Amyloidosis and monoclonal gammopathy of undetermined significance (MGUS patients contained FLCs. Furthermore MM and AL amyloidosis vesicles were strongly positive for Hsp70, annexin V, and c-src compared to MGUS and control patients. These are the first data implying that FLCs reroute via microvesicles in the blood stream, and also suggest a potential novel mechanism of c-src activation in plasma cell dyscrasia.

  2. Hysteretic behavior of special shaped columns composed of steel and reinforced concrete (SRC)

    Chen, Zongping; Xu, Jinjun; Xue, Jianyang


    This paper describes a series of experimental investigations on seventeen specimens of steel reinforced concrete special shaped (SRCSS) columns under low cyclic reversed loading using parallel crosshead equipment. Nine T-shaped SRC columns, four L-shaped SRC columns and four +-shaped SRC columns were tested to examine the effects of shape steel configuration, loading angle, axial compressive ratio and shear-span ratio on the behavior (strength, stiffness, energy dissipation, ductility, etc.) of SRCSS column specimens. The failure modes and hysteretic performance of all the specimens were obtained in the tests. Test results demonstrate that the shear-span ratio is the main parameter affecting the failure modes of SRCSS columns. The specimens with small shear-span ratio are prone to shear failure, and the primary failure planes in SRCSS columns are parallel to the loading direction. As a result, there is a symmetry between positive and negative loading directions in the hysteretic curves of the SRCSS columns. The majority of displacement ductility coefficients for all the specimens are over 3.0, so that the SRCSS columns demonstrate a better deformation capacity. In addition, the equivalent viscous damping coefficients of all the specimens are greater than 0.2, indicating that the seismic behavior of SRCSS columns is adequate. Finally, the superposition theory was used to calculate the limits of axial compressive ratio for the specimens, and it is found that the test axial compressive ratio is close to or smaller than the calculated axial compressive ratio limit.

  3. SRC-II slurry preheater technical uncertainties. Report for the technical data analysis program


    This report reviews the performance, and draws conclusions therefrom, the coal slurry preheaters of the Ft. Lewis, Washington, Solvent Refined Coal (SRC) Pilot Plant in the following areas: Coking, Erosion Corrosion, Heat transfer and pressure drop effects. Using prudent engineering judgement it postulates how such conclusions should affect the design and operability of large preheaters in future commercial scale plants. Also a recommendation is made for a small scale research and development effort that should result in a much firmer preheater design for any future facility. This report should be read in conjunction with the Solvent Refined Coal (SRC) Final Report, and volumes 1 and 2 of Slurry Preheater Design, SRC-II Process and also Ft. Lewis Slurry Preheater Data Analysis, 1-1/2 Inch Coil by Gulf Science and Technology Company of Pittsburgh, Pennsylvania. The Pittsburg and Midway Coal Mining Co.'s background is based primarily on a racetrack shaped up-flow coil and these comments pertain specifically to a commercial heater of that type of design. 5 references, 12 figures, 1 table.


    Parry, G.; Bartholomew, J.A.; Blssell, M.J.


    We report here a study of the mechanisms leading to loss of growth control in chicken embryo fibroblasts transformed by Rous sarcoma virus (RSV). We have been particularly concerned with the role of the src gene in this process, and have used RSV mutants temperature sensitive (ts) for transformation to investigate the nature of the growth regulatory lesion. The two principal findings were (1) the stationary phase of the cell cycle (G{sub 1}) in chick embryo fibroblasts seems to have two distinct regulatory compartments (using the terminology of Brooks et al. we refer to these as 'Q' and 'A' states). When rendered stationary at 41.5 C by serum deprivation, normal cells enter a Q state, but cells infected with the ts-mutant occupy an A state. (2) Whereas normal cells can occupy either state depending on culture conditions, the ts-infected cells, at 41.5 C, do not seem to enter Q even though a known src gene product, a kinase, is reported to be inactive at this temperature. We discuss the possibility that viral factors other than the active src protein kinase influence growth control in infected cultures.

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

    Vladan Ivetić


    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.     

  6. Seamless Provenance Representation and Use in Collaborative Science Scenarios

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


    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

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

    Chi-Ming Wu


    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.

  8. D-pinitol Inhibits Prostate Cancer Metastasis through Inhibition of αVβ3 Integrin by Modulating FAK, c-Src and NF-κB Pathways

    Chih-Hsin Tang


    Full Text Available Prostate cancer is the most commonly diagnosed malignancy in men and shows a predilection for metastasis to the bone. D-pinitol, a 3-methoxy analogue of d-chiro-inositol, was identified as an active principle in soy foods and legumes, and it has been proven to induce tumor apoptosis and metastasis of cancer cells. In this study, we investigated the anti-metastasis effects of D-pinitol in human prostate cancer cells. We found that D-pinitol reduced the migration and the invasion of prostate cancer cells (PC3 and DU145 at noncytotoxic concentrations. Integrins are the major adhesive molecules in mammalian cells and have been associated with the metastasis of cancer cells. Treatment of prostate cancer cells with D-pinitol reduced mRNA and cell surface expression of αvβ3 integrin. In addition, D-pinitol exerted its inhibitory effects by reducing focal adhesion kinase (FAK phosphorylation, c-Src kinase activity and NF-kB activation. Thus, D-pinitol may be a novel anti-metastasis agent for the treatment of prostate cancer metastasis.

  9. D-pinitol inhibits prostate cancer metastasis through inhibition of αVβ3 integrin by modulating FAK, c-Src and NF-κB pathways.

    Lin, Tien-Huang; Tan, Tzu-Wei; Tsai, Tsung-Hsun; Chen, Chi-Cheng; Hsieh, Teng-Fu; Lee, Shang-Sen; Liu, Hsin-Ho; Chen, Wen-Chi; Tang, Chih-Hsin


    Prostate cancer is the most commonly diagnosed malignancy in men and shows a predilection for metastasis to the bone. D-pinitol, a 3-methoxy analogue of d-chiro-inositol, was identified as an active principle in soy foods and legumes, and it has been proven to induce tumor apoptosis and metastasis of cancer cells. In this study, we investigated the anti-metastasis effects of D-pinitol in human prostate cancer cells. We found that D-pinitol reduced the migration and the invasion of prostate cancer cells (PC3 and DU145) at noncytotoxic concentrations. Integrins are the major adhesive molecules in mammalian cells and have been associated with the metastasis of cancer cells. Treatment of prostate cancer cells with D-pinitol reduced mRNA and cell surface expression of αvβ3 integrin. In addition, D-pinitol exerted its inhibitory effects by reducing focal adhesion kinase (FAK) phosphorylation, c-Src kinase activity and NF-kB activation. Thus, D-pinitol may be a novel anti-metastasis agent for the treatment of prostate cancer metastasis.

  10. Targeting androgen receptor/Src complex impairs the aggressive phenotype of human fibrosarcoma cells.

    Gabriella Castoria

    Full Text Available BACKGROUND: Hormones and growth factors influence the proliferation and invasiveness of human mesenchymal tumors. The highly aggressive human fibrosarcoma HT1080 cell line harbors classical androgen receptor (AR that responds to androgens triggering cell migration in the absence of significant mitogenesis. As occurs in many human cancer cells, HT1080 cells also express epidermal growth factor receptor (EGFR. EXPERIMENTAL: FINDINGS: We report that the pure anti-androgen Casodex inhibits the growth of HT1080 cell xenografts in immune-depressed mice, revealing a novel role of AR in fibrosarcoma progression. In HT1080 cultured cells EGF, but not androgens, robustly increases DNA synthesis. Casodex abolishes the EGF mitogenic effect, implying a crosstalk between EGFR and AR. The mechanism underlying this crosstalk has been analyzed using an AR-derived small peptide, S1, which prevents AR/Src tyrosine kinase association and androgen-dependent Src activation. Present findings show that in HT1080 cells EGF induces AR/Src Association, and the S1 peptide abolishes both the assembly of this complex and Src activation. The S1 peptide inhibits EGF-stimulated DNA synthesis, cell matrix metalloproteinase-9 (MMP-9 secretion and invasiveness of HT1080 cells. Both Casodex and S1 peptide also prevent DNA synthesis and migration triggered by EGF in various human cancer-derived cells (prostate, breast, colon and pancreas that express AR. CONCLUSION: This study shows that targeting the AR domain involved in AR/Src association impairs EGF signaling in human fibrosarcoma HT1080 cells. The EGF-elicited processes inhibited by the peptide (DNA synthesis, MMP-9 secretion and invasiveness cooperate in increasing the aggressive phenotype of HT1080 cells. Therefore, AR represents a new potential therapeutic target in human fibrosarcoma, as supported by Casodex inhibition of HT1080 cell xenografts. The extension of these findings in various human cancer-derived cell lines

  11. The WW domain of Yes-associated protein binds a proline-rich ligand that differs from the consensus established for Src homology 3-binding modules.

    Chen, H I; Sudol, M


    The WW domain has previously been described as a motif of 38 semiconserved residues found in seemingly unrelated proteins, such as dystrophin, Yes-associated protein (YAP), and two transcriptional regulators, Rsp-5 and FE65. The molecular function of the WW domain has been unknown until this time. Using a functional screen of a cDNA expression library, we have identified two putative ligands of the WW domain of YAP, which we named WBP-1 and WBP-2. Peptide sequence comparison between the two partial clones revealed a homologous region consisting of a proline-rich domain followed by a tyrosine residue (with the shared sequence PPPPY), which we shall call the PY motif. Binding assays and site-specific mutagenesis have shown that the PY motif binds with relatively high affinity and specificity to the WW domain of YAP, with the preliminary consensus XPPXY being critical for binding. Herein, we have implicated the WW domain with a role in mediating protein-protein interactions, as a variant of the paradigm set by Src homology 3 domains and their proline-rich ligands.

  12. Provenance for Runtime Workflow Steering and Validation in Computational Seismology

    Spinuso, A.; Krischer, L.; Krause, A.; Filgueira, R.; Magnoni, F.; Muraleedharan, V.; David, M.


    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 ( 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

  13. Towards Provenance and Traceability in CRISTAL for HEP

    Shamdasani, Jetendr; McClatchey, Richard


    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.

  14. CD133/Src axis mediates tumor initiating property and epithelial-mesenchymal transition of head and neck cancer.

    Yu-Syuan Chen

    Full Text Available BACKGROUND: Head and Neck squamous cell carcinoma (HNSCC is a human lethal cancer with clinical, pathological, phenotypical and biological heterogeneity. Caner initiating cells (CICs, which are responsible for tumor growth and coupled with gain of epithelial-mesenchymal transition (EMT, have been identified. Previously, we enriched a subpopulation of head and neck cancer initiating cells (HN-CICs with up-regulation of CD133 and enhancement of EMT. Others demonstrate that Src kinase interacts with and phosphorylates the cytoplasmic domain of CD133. However, the physiological function of CD133/Src signaling in HNSCCs has not been uncovered. METHODOLOGY/PRINCIPAL FINDING: Herein, we determined the critical role of CD133/Src axis modulating stemness, EMT and tumorigenicity of HNSCC and HN-CICs. Initially, down-regulation of CD133 significantly reduced the self-renewal ability and expression of stemness genes, and promoted the differentiation and apoptotic capability of HN-CICs. Additionally, knockdown of CD133 in HN-CICs also lessened both in vitro malignant properties including cell migration/cell invasiveness/anchorage independent growth, and in vivo tumor growth by nude mice xenotransplantation assay. In opposite, overexpression of CD133 enhanced the stemness properties and tumorigenic ability of HNSCCs. Lastly, up-regulation of CD133 increased phosphorylation of Src coupled with EMT transformation in HNSCCs, on the contrary, silence of CD133 or treatment of Src inhibitor inversely abrogated above phenotypic effects, which were induced by CD133 up-regulation in HNSCCs or HN-CICs. CONCLUSION/SIGNIFICANCE: Our results suggested that CD133/Src signaling is a regulatory switch to gain of EMT and of stemness properties in HNSCC. Finally, CD133/Src axis might be a potential therapeutic target for HNSCC by eliminating HN-CICs.

  15. 4-Aryl-4H-chromene-3-carbonitrile derivatives: evaluation of Src kinase inhibitory and anticancer activities.

    Fallah-Tafti, Asal; Tiwari, Rakesh; Shirazi, Amir Nasrolahi; Akbarzadeh, Tahmineh; Mandal, Deendayal; Shafiee, Abbas; Parang, Keykavous; Foroumadi, Alireza


    Src kinase mutations and/or overexpression have been implicated in the development of a number of human cancer including colon, breast, and lung cancers. Thus, designing potent and selective Src kinase inhibitors as anticancer agents is a subject of major interest. A series of 4-aryl substituted derivatives of 2-amino-7-dimethylamino-4H-chromene-3-carbonitrile were synthesized using one-pot reaction of appropriate substituted aromatic aldehydes, malononitrile, and 3-(dimethylamino)phenol in the presence of piperidine. All 23 compounds were evaluated for inhibition of Src kinase and cell proliferation in human colon adenocarcinoma (HT-29) and leukemia (CCRF-CEM) cell lines. Among the tested compounds, 2-chlorophenyl- (4c), 3-nitrophenyl- (4h), 4-trifluoromethyphenyl- (4i), and 2,3-dichlorophenyl- (4k) substituted chromenes showed Src kinase inhibitory effect with IC(50) values of 11.1-18.3 µM. Compound 4c was relatively selective against Src (IC(50) = 11.1 µM), when compared with selected kinases, epidermal growth factor receptor (EGFR, IC(50) > 300 µM), C-terminal Src kinase (Csk, IC(50) = 101.7 µM), and lymphocyte-specific protein tyrosine kinase (Lck, IC(50) = 46.8 µM). 3-Chlorophenyl substituted thiazole (4v) and 2-chlorophenylsubstituted thiazole (4u) chromene derivatives inhibited the cell proliferation of HT-29 and CCRF-CEM by 80% and 50% respectively, at a concentration of 50 µM. The data indicate that 4H-chromene-3-carbonitrile scaffold has the potential to be optimized further for designing more potent Src kinase inhibitors and/or anticancer lead compounds.

  16. A novel 3D fibril force assay implicates src in tumor cell force generation in collagen networks.

    Robert J Polackwich

    Full Text Available New insight into the biomechanics of cancer cell motility in 3D extracellular matrix (ECM environments would significantly enhance our understanding of aggressive cancers and help identify new targets for intervention. While several methods for measuring the forces involved in cell-matrix interactions have been developed, previous to this study none have been able to measure forces in a fibrillar environment. We have developed a novel assay for simultaneously measuring cell mechanotransduction and motility in 3D fibrillar environments. The assay consists of a controlled-density fibrillar collagen gel atop a controlled-stiffness polyacrylamide (PAA surface. Forces generated by living cells and their migration in the 3D collagen gel were measured with the 3D motion of tracer beads within the PAA layer. Here, this 3D fibril force assay is used to study the role of the invasion-associated protein kinase Src in mechanotransduction and motility. Src expression and activation are linked with proliferation, invasion, and metastasis, and have been shown to be required in 2D for invadopodia membranes to direct and mediate invasion. Breast cancer cell line MDA-MD-231 was stably transfected with GFP-tagged constitutively active Src or wild-type Src. In 3D fibrillar collagen matrices we found that, relative to wild-type Src, constitutively active Src: 1 increased the strength of cell-induced forces on the ECM, 2 did not significantly change migration speed, and 3 increased both the duration and the length, but not the number, of long membrane protrusions. Taken together, these results support the hypothesis that Src controls invasion by controlling the ability of the cell to form long lasting cellular protrusions to enable penetration through tissue barriers, in addition to its role in promoting invadopodia matrix-degrading activity.

  17. The transcriptional coactivators p/CIP and SRC-1 control insulin resistance through IRS1 in obesity models.

    Zhiyong Wang

    Full Text Available Three p160 family members, p/CIP, SRC1, and TIF2, have been identified as transcriptional coactivators for nuclear hormone receptors and other transcription factors in vitro. In a previous study, we reported initial characterization of the obesity-resistant phenotypes of p/CIP and SRC-1 double knockout (DKO mice, which exhibit increased energy expenditure, and suggested that nuclear hormone receptor target genes were involved in these phenotypes. In this study, we demonstrate that p/CIP and SRC1 control insulin signaling in a cell-autonomous manner both in vitro and in vivo. Genetic deletion of p/CIP and SRC-1 increases glucose uptake and enhances insulin sensitivity in both regular chow- and high fat diet-fed DKO mice despite increased food intake. Interestingly, we discover that loss of p/CIP and SRC-1 results in resistance to age-related obesity and glucose intolerance. We show that expression levels of a key insulin signaling component, insulin receptor substrate 1 (IRS1, are significantly increased in two cell lines representing fat and muscle lineages with p/CIP and SRC-1 deletions and in white adipose tissue and skeletal muscle of DKO mice; this may account for increased glucose metabolism and insulin sensitivity. This is the first evidence that the p160 coactivators control insulin signaling and glucose metabolism through IRS1. Therefore, our studies indicate that p/CIP and SRC-1 are potential therapeutic targets not only for obesity but also for diabetes.

  18. A novel activating role of SRC and STAT3 on HGF transcription in human breast cancer cells

    Elliott Bruce E


    Full Text Available Abstract We have previously determined that the HGF promoter can be transactivated by a combination of activated Src and wild-type Stat3 in the mouse breast cell lines HC11 and SP1. To determine if this pathway is of relevance for the human disease, a series of human breast and other human cells lines were examined, and the status of key proteins in these cells determined. All of the human breast cell lines exhibited strong transactivation by a combination of activated Src and Stat3. This activation was dependent on a Stat3 recognition element present at nt-95. The exception was the ErbB2 over-expressing cell line SK-BR-3 where Stat3 alone could transactivate HGF though Src augmented this effect. Increased phosphorylation of Stat3 tyrosine 705 was also observed in this line. Analysis of three ovarian cell lines revealed that Src/Stat3 expression was not able to activate the HGF promoter in two of these lines (SKOV3 and IOSE-80PC. Src/Stat3 expression did activate HGF transcription in OVCAR3 cells, but this effect was not mediated by the Stat3 site at nt-95. Stat3 phosphorylation at tyrosine 705 was observed in IOSE-80PC cells, but was insufficient to allow for activation of the HGF promoter. Human kidney (HEK293 and cervical carcinoma (HeLa cells were also not Src/Stat3 permissive, despite high levels of Stat3 phospho-Y705. These results suggest that human breast cells are a uniquely permissive environment for HGF transactivation by Src/Stat3 which may allow for the inappropriate activation of HGF transcription during the early stages of breast transformation. This could lead to paracrine or autocrine activation of the Met receptor in breast carcinoma cells.


    Sh. Salehpour


    Full Text Available ObjectivePfeiffer Syndrome is as rare as Apert syndrome in the Western population. This condition is very rare in the Asian population. At the best of our knowledge this is the first genetically proven case report from Iran. The authors report with a review of literature, the case of a infant with Pfeiffer syndrome, manifested by Lacunar skull, ventriculomegaly, bicoronal craniosynostosis,frontal bossing, shallow orbits, parrot-like nose, umbilical hernia, broad and medially deviated great toes.

  20. Geological provenance of Quaternary deposits from the southeastern Brazilian coast

    Anjos, R.M. [Instituto de Fisica, Universidade Federal Fluminense, Av. Gal. Milton Tavares de Souza s/n, Gragoata, Campus da Praia Vermelha, 24210-346, Niteroi, R.J. (Brazil)]. E-mail:; Veiga, R. [Instituto de Fisica, Universidade Federal Fluminense, Av. Gal. Milton Tavares de Souza s/n, Gragoata, Campus da Praia Vermelha, 24210-346, Niteroi, R.J. (Brazil); Carvalho, C. [Instituto de Fisica, Universidade Federal Fluminense, Av. Gal. Milton Tavares de Souza s/n, Gragoata, Campus da Praia Vermelha, 24210-346, Niteroi, R.J. (Brazil); Macario, K.D. [Instituto de Fisica, Universidade Federal Fluminense, Av. Gal. Milton Tavares de Souza s/n, Gragoata, Campus da Praia Vermelha, 24210-346, Niteroi, R.J. (Brazil); Gomes, P.R.S. [Instituto de Fisica, Universidade Federal Fluminense, Av. Gal. Milton Tavares de Souza s/n, Gragoata, Campus da Praia Vermelha, 24210-346, Niteroi, R.J. (Brazil)


    Natural gamma radiation measurements of beach sand deposits were performed with the aim of understanding the provenance and transport processes of sediments along the coastal zone of three Brazilian States. The method employs thorium, uranium and potassium as tracers of the mineralogical properties of beach sand minerals, which reflect the geological history of transport and sorting processes. A considerable positive correlation with the geological evolution of these Quaternary coastal deposits was observed.

  1. Biopsy proven acute interstitial nephritis after treatment with moxifloxacin


    Abstract Background Acute interstitial nephritis (AIN) is an important cause of reversible acute kidney injury. At least 70% of AIN is caused by various drugs, mainly penicillines and non-steroidal anti-inflammatory drugs. Quinolones are only rarely known to cause AIN and so far cases have been mainly described with older fluoroquinolones. Case Presentation Here we describe a case of biopsy proven interstitial nephritis after moxifloxacin treatment. The patient presented with fever, rigors an...

  2. Provenance Representation in the Global Change Information System (GCIS)

    Tilmes, Curt


    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.

  3. Role of intravenous immunoglobulin in suspected or proven neonatal sepsis



    Neonatal sepsis remains the major cause of mortality and morbidity including neurodevelopmental impairment and prolonged hospital stay in newborn infants .Despite of advances in technology and optimal antibiotic tre-atment, incidence of neonatal sepsis and its complications remains unacceptably high especially in developing countries .Premature neonates in particular are at higher risk due to developmentally immature host defence mecha-nisms.Though not approved by Food and Drug Administration ( FDA ) U.S.A, off label use of intravenous immunoglobulin as prophylactic or adjuvant agent in suspected or proven neonatal infections continues in many countries.In a recent large multicenter clinical trial by International Neonatal Immunotherapy Study (INIS) group, the use of polyvalent IgG immune globulin was not associated with significant differences in the risk of major com -plications or other adverse outcomes in neonates with suspected or proven sepsis .Hence, use of intravenous immu-noglobulin in suspected or proven neonatal sepsis is not recommended .The expense of prophylactic use of intrave-nous immunoglobulin administration for both term and preterm newborn population , given the minimal benefit as demonstrated by many individual studies and by meta-analysis is not justified .

  4. Quantifying the provenance of aeolian sediments using multiple composite fingerprints

    Liu, Benli; Niu, Qinghe; Qu, Jianjun; Zu, Ruiping


    We introduce a new fingerprinting method that uses multiple composite fingerprints for studies of aeolian sediment provenance. We used this method to quantify the provenance of sediments on both sides of the Qinghai-Tibetan Railway (QTR) in the Cuona Lake section of the Tibetan Plateau (TP), in an environment characterized by aeolian and fluvial interactions. The method involves repeatedly solving a linear mixing model based on mass conservation; the model is not limited to spatial scale or transport types and uses all the tracer groups that passed the range check, Kruskal-Wallis H-test, and a strict analytical solution screening. The proportional estimates that result from using different composite fingerprints are highly variable; however, the average of these fingerprints has a greater accuracy and certainty than any single fingerprint. The results show that sand from the lake beach, hilly surface, and gullies contribute, respectively, 48%, 31% and 21% to the western railway sediments and 43%, 33% and 24% to the eastern railway sediments. The difference between contributions from various sources on either side of the railway, which may increase in the future, was clearly related to variations in local transport characteristics, a conclusion that is supported by grain size analysis. The construction of the QTR changed the local cycling of materials, and the difference in provenance between the sediments that are separated by the railway reflects the changed sedimentary conditions on either side of the railway. The effectiveness of this method suggests that it will be useful in other studies of aeolian sediments.

  5. Incidence of malignancies in biopsy-proven inflammatory myopathy

    Meena A Kannan


    Full Text Available Background: Inflammatory myopathy (IM as a manifestation of paraneoplastic syndrome has been well-documented. However, the prevalence of malignancies reported varies across the studies. There are very few studies reported from Asia, only one from India. Aim: The aim of this analysis was to study the prevalence of malignancy in biopsy-proven cases of IM in India and to study the difference between malignant and non-malignant groups. Materials and Methods: The study was a retrospective review of case records of patients with a biopsy-proven IM attending Tertiary Care University Hospital. Results: Of the total 86 patients with biopsy-proven IM, 22 patients were polymyositis, 63 patients had dermatomyositis (DM and one was with an inclusion body myositis, not included for further analysis. Associated malignancy was diagnosed in 6 (7% patients, and five of them were females. Diagnosis of associated malignancy was identified at the time of diagnosis of IM in four (66.7% patients. All the six patients with an associated malignancy had DM. Only one patient died within 1 year of diagnosis. Creatinine kinase was much lower in patients with malignancy associated IM than in patients with no malignancy (P < 0.0001. Conclusion: The prevalence of malignancy was very low in our cohort as compared to the studies from other countries. Breast cancer was the most common malignancy associated with DM. The type of associated malignancy was quite variable.

  6. A Provenance Model for Real-Time Water Information Systems

    Liu, Q.; Bai, Q.; Zednik, S.; Taylor, P.; Fox, P. A.; Taylor, K.; Kloppers, C.; Peters, C.; Terhorst, A.; West, P.; Compton, M.; Shu, Y.; Provenance Management Team


    Generating hydrological data products, such as flow forecasts, involves complex interactions among instruments, data simulation models, computational facilities and data providers. Correct interpretation of the data produced at various stages requires good understanding of how data was generated or processed. Provenance describes the lineage of a data product. Making provenance information accessible to hydrologists and decision makers not only helps to determine the data’s value, accuracy and authorship, but also enables users to determine the trustworthiness of the data product. In the water domain, WaterML2 [1] is an emerging standard which describes an information model and format for the publication of water observations data in XML. The W3C semantic sensor network incubator group (SSN-XG) [3] is producing ontologies for the description of sensor configurations. By integrating domain knowledge of this kind into the provenance information model, the integrated information model will enable water domain researchers and water resource managers to better analyse how observations and derived data products were generated. We first introduce the Proof Mark Language (PML2) [2], WaterML2 and the SSN-XG sensor ontology as the proposed provenance representation formalism. Then we describe some initial implementations how these standards could be integrated to represent the lineage of water information products. Finally we will highlight how the provenance model for a distributed real-time water information system assists the interpretation of the data product and establishing trust. Reference [1] Taylor, P., Walker, G., Valentine, D., Cox, Simon: WaterML2.0: Harmonising standards for water observation data. Geophysical Research Abstracts. Vol. 12. [2] da Silva, P.P., McGuinness, D.L., Fikes, R.: A proof markup language for semantic web services. Inf. Syst. 31(4) (2006), 381-395. [3] W3C Semantic Sensor Network Incubator Group

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

    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.

  8. Src64 controls a novel actin network required for proper ring canal formation in the Drosophila male germline.

    Eikenes, Åsmund Husabø; Malerød, Lene; Lie-Jensen, Anette; Sem Wegner, Catherine; Brech, Andreas; Liestøl, Knut; Stenmark, Harald; Haglund, Kaisa


    In many organisms, germ cells develop as cysts in which cells are interconnected via ring canals (RCs) as a result of incomplete cytokinesis. However, the molecular mechanisms of incomplete cytokinesis remain poorly understood. Here, we address the role of tyrosine phosphorylation of RCs in the Drosophila male germline. We uncover a hierarchy of tyrosine phosphorylation within germline cysts that positively correlates with RC age. The kinase Src64 is responsible for mediating RC tyrosine phosphorylation, and loss of Src64 causes a reduction in RC diameter within germline cysts. Mechanistically, we show that Src64 controls an actin network around the RCs that depends on Abl and the Rac/SCAR/Arp2/3 pathway. The actin network around RCs is required for correct RC diameter in cysts of developing germ cells. We also identify that Src64 is required for proper germ cell differentiation in the Drosophila male germline independent of its role in RC regulation. In summary, we report that Src64 controls actin dynamics to mediate proper RC formation during incomplete cytokinesis during germline cyst development in vivo.

  9. Antioxidants decrease the apoptotic effect of 5-Fu in colon cancer by regulating Src-dependent caspase-7 phosphorylation

    Fu, Y; Yang, G; Zhu, F; Peng, C; Li, W; Li, H; Kim, H-G; Bode, A M; Dong, Z; Dong, Z


    Although the rate of development of drug resistance remains very high, 5-fluorouracil (5-Fu) is still the most common chemotherapeutic drug used for the treatment of colon cancer. A better understanding of the mechanism of why cancers develop resistance to 5-Fu could improve its therapeutic effect. Sometimes, antioxidants are used simultaneously with 5-Fu treatment. However, a recent clinical trial showed no advantage or even a harmful effect of combining antioxidants with 5-Fu compared with administration of 5-Fu alone. The mechanism explaining this phenomenon is still poorly understood. In this study, we show that 5-Fu can induce reactive oxygen species-dependent Src activation in colon cancer cells. Mouse embryonic fibroblasts that are deficient in Src showed a clear resistance to 5-Fu, and knocking down Src protein expression in colon cancer cells also decreased 5-Fu-induced apoptosis. We found that Src could interact with and phosphorylate caspase-7 at multiple tyrosine sites. Functionally, the tyrosine phosphorylation of caspase-7 increases its activity, thereby enhancing cellular apoptosis. When using 5-Fu and antioxidants together, Src activation was blocked, resulting in decreased 5-Fu-induced apoptosis. Our results provide a novel explanation as to why 5-Fu is not effective in combination with some antioxidants in colon cancer patients, which is important for clinical chemotherapy. PMID:24407236

  10. The role(s) of Src kinase and Cbl proteins in the regulation of osteoclast differentiation and function.

    Horne, William C; Sanjay, Archana; Bruzzaniti, Angela; Baron, Roland


    The osteoclast resorbs mineralized bone during bone development, homeostasis, and repair. The deletion of the gene encoding the nonreceptor tyrosine kinase c-Src produces an osteopetrotic skeletal phenotype that is the consequence of the inability of the mature osteoclast to efficiently resorb bone. Src-/- osteoclasts exhibit reduced motility and abnormal organization of the apical secretory domain (the ruffled border) and attachment-related cytoskeletal elements that are necessary for bone resorption. A key function of Src in osteoclasts is to promote the rapid assembly and disassembly of the podosomes, the specialized integrin-based attachment structures of osteoclasts and other highly motile cells. Once recruited to the activated integrins, especially alphavbeta3), by the adhesion tyrosine kinase Pyk2, Src binds and phosphorylates Cbl and Cbl-b, homologous multisite adapter proteins with ubiquitin ligase activity. The Cbl proteins in turn recruit and activate additional signaling effectors, including phosphatidylinositol 3-kinase and dynamin, which play key roles in the development of cell polarity and the regulation of cell attachment and motility. In addition, Src and the Cbl proteins contribute to signaling cascades that are activated by several important receptors, including receptor activator of nuclear factor kappaB and the macrophage colony-stimulating factor receptor, and also downregulate the signaling from many of these receptors.

  11. SRC burn test in 700-hp oil-designed boiler. Volume 1. Integrated report. Final technical report


    This burn test program was conducted during the period of August 1982 to February 1983 to demonstrate that Solvent Refined Coal (SRC) products can displace petroleum as a boiler fuel in oil- and gas-designed boilers. The test program was performed at the U.S. Department of Energy's Pittsburgh Energy Technology Center (PETC). Three forms of SRC (pulverized SRC, a solution of SRC dissolved in process-derived distillates, and a slurry of SRC and water) and No. 6 Fuel Oil were evaluated in the 700-hp (30 x 10/sup 6/ Btu/hour) watertube, oil-designed boiler facility at PETC. The test program was managed by the International Coal Refining Company (ICRC) and sponsored by the Department of Energy. Other organizations were involved as necessary to provide the expertise required to execute the test program. This final report represents an integrated overview of the test program conducted at PETC. More detailed information with preliminary data can be obtained from separate reports prepared by PETC, Southern Research Institute, Wheelabrator-Frye, Babcock and Wilcox, and Combustion Engineering. These are presented as Annex Volumes A-F. 25 references, 41 figures, 15 tables.

  12. The inner nuclear membrane protein Src1 associates with subtelomeric genes and alters their regulated gene expression.

    Grund, Stefanie E; Fischer, Tamás; Cabal, Ghislain G; Antúnez, Oreto; Pérez-Ortín, José E; Hurt, Ed


    Inner nuclear membrane proteins containing a LEM (LAP2, emerin, and MAN1) domain participate in different processes, including chromatin organization, gene expression, and nuclear envelope biogenesis. In this study, we identify a robust genetic interaction between transcription export (TREX) factors and yeast Src1, an integral inner nuclear membrane protein that is homologous to vertebrate LEM2. DNA macroarray analysis revealed that the expression of the phosphate-regulated genes PHO11, PHO12, and PHO84 is up-regulated in src1Delta cells. Notably, these PHO genes are located in subtelomeric regions of chromatin and exhibit a perinuclear location in vivo. Src1 spans the nuclear membrane twice and exposes its N and C domains with putative DNA-binding motifs to the nucleoplasm. Genome-wide chromatin immunoprecipitation-on-chip analyses indicated that Src1 is highly enriched at telomeres and subtelomeric regions of the yeast chromosomes. Our data show that the inner nuclear membrane protein Src1 functions at the interface between subtelomeric gene expression and TREX-dependent messenger RNA export through the nuclear pore complexes.

  13. Rescue of a trafficking defective human pacemaker channel via a novel mechanism: roles of Src, Fyn, and Yes tyrosine kinases.

    Lin, Yen-Chang; Huang, Jianying; Kan, Hong; Frisbee, Jefferson C; Yu, Han-Gang


    Therapeutic strategies such as using channel blockers and reducing culture temperature have been used to rescue some long QT-associated voltage-gated potassium Kv trafficking defective mutant channels. A hyperpolarization-activated cyclic nucleotide-gated HCN4 pacemaker channel mutant (D553N) has been recently found in a patient associated with cardiac arrhythmias including long QT. D553N showed the defective trafficking to the cell surface, leading to little ionic current expression (loss-of-function). We show in this report that enhanced tyrosine phosphorylation mediated by Src, Fyn, and Yes kinases was able to restore the surface expression of D553N for normal current expression. Src or Yes, but not Fyn, significantly increased the current density and surface expression of D553N. Fyn accelerated the activation kinetics of the rescued D553N. Co-expression of D553N with Yes exhibited the slowest activation kinetics of D553N. Src, Fyn, and Yes significantly enhanced the tyrosine phosphorylation of D553N. A combination of Src, Fyn, and Yes rescued the current expression and the gating of D553N comparable with those of wild-type HCN4. In conclusion, we demonstrate a novel mechanism using three endogenous Src kinases to rescue a trafficking defective HCN4 mutant channel (D553N) by enhancing the tyrosine phosphorylation of the mutant channel protein.

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

    Sancier, Florence; Dumont, Aurélie; Sirvent, Audrey; Paquay de Plater, Ludmilla; Edmonds, Thomas; David, Géraldine; Jan, Michel; de Montrion, Catherine; Cogé, Francis; Léonce, Stéphane; Burbridge, Michael; Bruno, Alain; Boutin, Jean A; Lockhart, Brian; Roche, Serge; Cruzalegui, Francisco


    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.

  15. Chemopreventive activity of plant flavonoid isorhamnetin in colorectal cancer is mediated by oncogenic Src and β-catenin.

    Saud, Shakir M; Young, Matthew R; Jones-Hall, Yava L; Ileva, Lilia; Evbuomwan, Moses O; Wise, Jennifer; Colburn, Nancy H; Kim, Young S; Bobe, Gerd


    Analysis of the Polyp Prevention Trial showed an association between an isorhamnetin-rich diet and a reduced risk of advanced adenoma recurrence; however, the mechanism behind the chemoprotective effects of isorhamnetin remains unclear. Here, we show that isorhamnetin prevents colorectal tumorigenesis of FVB/N mice treated with the chemical carcinogen azoxymethane and subsequently exposed to colonic irritant dextran sodium sulfate (DSS). Dietary isorhamnetin decreased mortality, tumor number, and tumor burden by 62%, 35%, and 59%, respectively. MRI, histopathology, and immunohistochemical analysis revealed that dietary isorhamnetin resolved the DSS-induced inflammatory response faster than the control diet. Isorhamnetin inhibited AOM/DSS-induced oncogenic c-Src activation and β-catenin nuclear translocation, while promoting the expression of C-terminal Src kinase (CSK), a negative regulator of Src family of tyrosine kinases. Similarly, in HT-29 colon cancer cells, isorhamnetin inhibited oncogenic Src activity and β-catenin nuclear translocation by inducing expression of csk, as verified by RNA interference knockdown of csk. Our observations suggest the chemoprotective effects of isorhamnetin in colon cancer are linked to its anti-inflammatory activities and its inhibition of oncogenic Src activity and consequential loss of nuclear β-catenin, activities that are dependent on CSK expression.

  16. Roles of mitochondrial Src tyrosine kinase and zinc in nitric oxide-induced cardioprotection against ischemia/reperfusion injury.

    Zhang, Y; Xing, F; Zheng, H; Xi, J; Cui, X; Xu, Z


    While nitric oxide (NO) induces cardioprotection by targeting the mitochondrial permeability transition pore (mPTP), the precise mitochondrial signaling events that mediate the action of NO remain unclear. The purpose of this study was to test whether NO induces cardioprotection against ischemia/reperfusion by inhibiting oxidative stress through mitochondrial zinc and Src tyrosine kinase. The NO donor S-nitroso-N-acetyl penicillamine (SNAP) given before the onset of ischemia reduced cell death in rat cardiomyocytes subjected to simulated ischemia/reperfusion, and this was abolished by the zinc chelator N,N,N',N'-tetrakis-(2-pyridylmethyl)ethylenediamine (TPEN) and the Src tyrosine kinase inhibitor PP2. SNAP also prevented loss of mitochondrial membrane potential (ΔΨm) at reperfusion, an effect that was blocked by TPEN and PP2. SNAP increased mitochondrion-free zinc upon reperfusion and enhanced mitochondrial Src phosphorylation in a zinc-dependent manner. SNAP inhibited both mitochondrial complex I activity and mitochondrial reactive oxygen species (ROS) generation at reperfusion through zinc and Src tyrosine kinase. Finally, the anti-infarct effect of SNAP was abrogated by TPEN and PP2 applied at reperfusion in isolated rat hearts. In conclusion, NO induces cardioprotection at reperfusion by targeting mitochondria through attenuation of oxidative stress resulted from the inhibition of complex I at reperfusion. Activation of mitochondrial Src tyrosine kinase by zinc may account for the inhibition of complex I.

  17. Unlocking Doors without Keys: Activation of Src by Truncated C-terminal Intracellular Receptor Tyrosine Kinases Lacking Tyrosine Kinase Activity

    Belén Mezquita


    Full Text Available One of the best examples of the renaissance of Src as an open door to cancer has been the demonstration that just five min of Src activation is sufficient for transformation and also for induction and maintenance of cancer stem cells [1]. Many tyrosine kinase receptors, through the binding of their ligands, become the keys that unlock the structure of Src and activate its oncogenic transduction pathways. Furthermore, intracellular isoforms of these receptors, devoid of any tyrosine kinase activity, still retain the ability to unlock Src. This has been shown with a truncated isoform of KIT (tr-KIT and a truncated isoform of VEGFR-1 (i21-VEGFR-1, which are intracellular and require no ligand binding, but are nonetheless able to activate Src and induce cell migration and invasion of cancer cells. Expression of the i21-VEGFR-1 is upregulated by the Notch signaling pathway and repressed by miR-200c and retinoic acid in breast cancer cells. Both Notch inhibitors and retinoic acid have been proposed as potential therapies for invasive breast cancer.

  18. Unlocking Doors without Keys: Activation of Src by Truncated C-terminal Intracellular Receptor Tyrosine Kinases Lacking Tyrosine Kinase Activity.

    Mezquita, Belén; Mezquita, Pau; Pau, Montserrat; Mezquita, Jovita; Mezquita, Cristóbal


    One of the best examples of the renaissance of Src as an open door to cancer has been the demonstration that just five min of Src activation is sufficient for transformation and also for induction and maintenance of cancer stem cells [1]. Many tyrosine kinase receptors, through the binding of their ligands, become the keys that unlock the structure of Src and activate its oncogenic transduction pathways. Furthermore, intracellular isoforms of these receptors, devoid of any tyrosine kinase activity, still retain the ability to unlock Src. This has been shown with a truncated isoform of KIT (tr-KIT) and a truncated isoform of VEGFR-1 (i21-VEGFR-1), which are intracellular and require no ligand binding, but are nonetheless able to activate Src and induce cell migration and invasion of cancer cells. Expression of the i21-VEGFR-1 is upregulated by the Notch signaling pathway and repressed by miR-200c and retinoic acid in breast cancer cells. Both Notch inhibitors and retinoic acid have been proposed as potential therapies for invasive breast cancer.

  19. Src, a Molecular Switch Governing Gain Control of Synaptic Transmission Mediated by N-methyl-D-Aspartate Receptors

    Yu, Xian-Min; Salter, Michael W.


    The N-methyl-D-aspartate (NMDA) receptor is a principal subtype of glutamate receptor mediating fast excitatory transmission at synapses in the dorsal horn of the spinal cord and other regions of the central nervous system. NMDA receptors are crucial for the lasting enhancement of synaptic transmission that occurs both physiologically and in pathological conditions such as chronic pain. Over the past several years, evidence has accumulated indicating that the activity of NMDA receptors is regulated by the protein tyrosine kinase, Src. Recently it has been discovered that, by means of up-regulating NMDA receptor function, activation of Src mediates the induction of the lasting enhancement of excitatory transmission known as long-term potentiation in the CA1 region of the hippocampus. Also, Src has been found to amplify the up-regulation of NMDA receptor function that is produced by raising the intracellular concentration of sodium. Sodium concentration increases in neuronal dendrites during high levels of firing activity, which is precisely when Src becomes activated. Therefore, we propose that the boost in NMDA receptor function produced by the coincidence of activating Src and raising intracellular sodium may be important in physiological and pathophysiological enhancement of excitatory transmission in the dorsal horn of the spinal cord and elsewhere in the central nervous system.

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

    Huang, Chunfa, E-mail: [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)


    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.

  1. PSM/SH2B1 splice variants: critical role in src catalytic activation and the resulting STAT3s-mediated mitogenic response.

    Zhang, Manchao; Deng, Youping; Riedel, Heimo


    A role of PSM/SH2B1 had been shown in mitogenesis and extending to phenotypic cell transformation, however, the underlying molecular mechanism remained to be established. Here, four alternative PSM splice variants and individual functional protein domains were compared for their role in the regulation of Src activity. We found that elevated cellular levels of PSM variants resulted in phenotypic cell transformation and potentiated cell proliferation and survival in response to serum withdrawal. PSM variant activity presented a consistent signature pattern for any tested response of highest activity observed for gamma, followed by delta, alpha, and beta with decreasing activity. PSM-potentiated cell proliferation was sensitive to Src inhibitor herbimycin and PSM and Src were found in the same immune complex. PSM variants were substrates of the Src Tyr kinase and potentiated Src catalytic activity by increasing the V(max) and decreasing the K(m) for ATP with the signature pattern of variant activity. Dominant-negative PSM peptide mimetics including the SH2 or PH domains inhibited Src catalytic activity as well as Src-mediated phenotypic cell transformation. Activation of major Src substrate STAT3 was similarly potentiated by the PSM variants in a Src-dependent fashion or inhibited by PSM domain-specific peptide mimetics. Expression of a dominant-negative STAT3 mutant blocked PSM variant-mediated phenotypic cell transformation. Our results implicate an essential role of the PSM variants in the activation of the Src kinase and the resulting mitogenic response--extending to phenotypic cell transformation and involving the established Src substrate STAT3.

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


    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.

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

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


    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.

  4. Mutagenic and chemical properties of SRC-I materials: a status report

    Pelroy, R.A. (ed.)


    Process solvent (PS) and solid product (dissolved in organic solvent) produced by the Solvent Refined Coal (SRC-I) process at the Wilsonville pilot plant contain mutagenically active components against the Ames-Salmonella strains, Salmonella typhimurium TA98 and TA100. Ames-positive mutagens are apparently concentrated in the moderately polar to strongly polar chemical constituents of both materials. However, the dissolved solid product contains a much higher proportion of very strongly polar mutagens, and may be enriched in acidic - i.e., oxygen-containing mutagens.

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

    Bouzinova, Elena

    ,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...... of ouabain. Here we demonstrate results of our research on the mechanisms involved in the modulation of vascular wall contractility by ouabain-sensitive Na,K-ATPase. Methods: The experiments were performed using rat mesenteric arteries in isometric myograph conditions. To inhibit kinase activity a Src-family...

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

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


    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.

  7. Experimental Study on Elastic-Plastic Behavior of SRC Columns with High Strength Steel


    The demand to use high strength and high performance material because of large span and high rise of building in recent years. As to use of high-strength steel in composite steel and reinforced concrete structures, it remains to be clarified whether the ductile behavior can be ensured, especially when the high-strength steel is used in combination with High-strength concrete. This paper describes the test results on the elasto-plastic behavior of SRC column using high strength steel, and disc...

  8. The Results of ESO/SRC Plates Survey of Southern Hemisphere

    Gyulbudaghian, A. L.


    We have searched the ESO/SRC (E, B, R, J) charts of Southern Hemisphere for discovering of new star forming regions, cometary nebulae, HH objects, tight trapezium like systems, consisting of late type dwarf stars, jets from the stars, and also of radial systems of dark globules. This work was done in several places: in Estonia (Tartu observatory), in Mexico (Mexico, UNAM, Institute of Astronomy), in Chile (twice: ESO Vitacura, Cerro Calan Observatory, Santiago). As a result of this work several dozens of each mentioned above type of objects were found.

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

    Yeh-Chuin Poh

    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

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

    Nikolett Marton


    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.

  11. Provenance-Powered Automatic Workflow Generation and Composition

    Zhang, J.; Lee, S.; Pan, L.; Lee, T. J.


    In recent years, scientists have learned how to codify tools into reusable software modules that can be chained into multi-step executable workflows. Existing scientific workflow tools, created by computer scientists, require domain scientists to meticulously design their multi-step experiments before analyzing data. However, this is oftentimes contradictory to a domain scientist's daily routine of conducting research and exploration. We hope to resolve this dispute. Imagine this: An Earth scientist starts her day applying NASA Jet Propulsion Laboratory (JPL) published climate data processing algorithms over ARGO deep ocean temperature and AMSRE sea surface temperature datasets. Throughout the day, she tunes the algorithm parameters to study various aspects of the data. Suddenly, she notices some interesting results. She then turns to a computer scientist and asks, "can you reproduce my results?" By tracking and reverse engineering her activities, the computer scientist creates a workflow. The Earth scientist can now rerun the workflow to validate her findings, modify the workflow to discover further variations, or publish the workflow to share the knowledge. In this way, we aim to revolutionize computer-supported Earth science. We have developed a prototyping system to realize the aforementioned vision, in the context of service-oriented science. We have studied how Earth scientists conduct service-oriented data analytics research in their daily work, developed a provenance model to record their activities, and developed a technology to automatically generate workflow starting from user behavior and adaptability and reuse of these workflows for replicating/improving scientific studies. A data-centric repository infrastructure is established to catch richer provenance to further facilitate collaboration in the science community. We have also established a Petri nets-based verification instrument for provenance-based automatic workflow generation and recommendation.

  12. Delphinella Shoot Blight on Abies lasiocarpa Provenances in Norway

    Venche Talgø


    Full Text Available Delphinella shoot blight (Delphinella abietis attacks true firs (Abies spp. in Europe and North America. Especially subalpine fir (A. lasiocarpa, one of the main Christmas tree species in Norway, is prone to the disease. The fungus kills current year needles, and in severe cases entire shoots. Dead needles become covered with black fruiting bodies, both pycnidia and pseudothecia. Delphinella shoot blight has mainly been a problem in humid, coastal regions in the northwestern part of Southern Norway, but, probably due to higher precipitation in inland regions during recent years, heavy attacks were found in 2011 in a field trial with 76 provenances of subalpine fir in Southeastern Norway. However, the amount of precipitation seemed less important once the disease had established in the field. Significant differences in susceptibility between provenances were observed. In general, the more bluish the foliage was, the healthier the trees appeared. The analysis of provenance means indicated that, at least for the southern range, the disease ratings were correlated with foliage color. This study also includes isolation, identification, a pathogenicity test, a seed test and electron microscopy of the wax layer on the needles. The fungus was identified based on the morphology of spores and by sequencing the Internal Transcribed Spacer (ITS regions of the ribosomal DNA. Koch’s postulates were fulfilled. The fungus was found present on newly harvested seeds and may therefore spread via international seed trade. When comparing the wax layers on green and blue needles, those of the latter were significantly thicker, a factor that may be involved in disease resistance.


    Sh. Salehpour


    Full Text Available ObjectivePfeiffer Syndrome is as rare as Apert syndrome in the Western population. This condition is very rare in the Asian population. At the best of our knowledge this is the first genetically proven case report from Iran. The authors report with a review of literature, the case of a infant with Pfeiffer syndrome, manifested by Lacunar skull, ventriculomegaly, bicoronal craniosynostosis,frontal bossing, shallow orbits, parrot-like nose, umbilical hernia, broad and medially deviated great toes.Keywords: Acrocephalosyndactylia, Craniosynostoses, Broad and great toes, Pfeiffer, Syndrome

  14. Selections of Proven Medical Records in Acupuncture: Part Ⅰ

    YANG Jie-bin; XIAO Yuan-chun


    @@ Editor's Note: Pro. YANG Jie-bin specializes in acupuncture and herbology in treating diseases of internal medincine, gynecology, pediatrics and dermatology. He treats difficult and stubborn diseases through meridian differentiation of pathogenesis, treating pain syndrome by unblocking meridians and harmonizing qi and blood,treating acute and chronic stomach diseases from regulatingthe liver and spleen, treating liver and gallbladder diseases by discharging the liver and clearing the gallbladder, all with excellent effects. From this issue,we will publish his some proven medical records in acupuncture.

  15. Diagenesis, provenance and depositional environments of the Bunter Sandstone Formation

    Olivarius, Mette; Weibel, Rikke; Friis, Henrik

    The Bunter Sandstone Formation in the northern North German Basin has large geothermal potential with high porosity and permeability (generally >15% and >100 mD, respectively) and with pore fluid temperatures that are adequate for geothermal energy production (c. 55–60˚C). A combined investigation...... of diagenesis, provenance and depositional environments is used to identify the reservoir rocks that possess the best quality. This is accomplished by integrating various methods including: seismic reflection data, sedimentological description of cores, mineral quantification by point counting, measurement...

  16. From static to dynamic provenance analysis-Sedimentary petrology upgraded

    Garzanti, Eduardo


    The classical approach to sandstone petrology, established in the golden years of plate tectonics and based on the axiom that "detrital modes of sandstone suites primarily reflect the different tectonic settings of provenance terranes," has represented a benchmark for decades. The composition of sand and sandstone, however, simply provides us with a distorted image of the lithological structure of source terranes and gives us little clue whether they are allochthonous or autochthonous, orogenic or anorogenic, young or old. What we may able to see reflected in detrital modes is the nature of source terranes (continental, arc, oceanic) and the tectonostratigraphic level reached by erosion in space and time. The proposed new approach to the petrology of sand and sandstone (1) starts with a simple classification scheme circulated since the 1960s, which is purely descriptive, objective, and free of ill-defined ambiguous terms and (2) focuses on the nature and tectonostratigraphic level of source terranes. Further steps are essential to upgrade provenance analysis. Acquiring knowledge from modern settings is needed to properly identify and wherever possible correct for physical and chemical processes introducing environmental and diagenetic bias and thus address nature's complexities with adequate conceptual tools. Equally important is the integration of multiple techniques, ideally including bulk-sediment, multi-mineral, and single-mineral methods. Bulk-sediment petrography remains the fundamental approach that allows us to capture the most precious source of direct provenance information, represented by the mineralogy and texture of rock fragments. Bulk-sediment geochemistry, applicable also to silt and clay carried in suspension, is a superior method to check for hydraulic sorting, chemical weathering, and fertility of detrital minerals in different sediment sources. Detrital geochronology, thermochronology, and isotope geochemistry reveal the diverse time structures

  17. Fatty Acid Oxidation-Driven Src Links Mitochondrial Energy Reprogramming and Oncogenic Properties in Triple-Negative Breast Cancer

    Jun Hyoung Park


    Full Text Available Transmitochondrial cybrids and multiple OMICs approaches were used to understand mitochondrial reprogramming and mitochondria-regulated cancer pathways in triple-negative breast cancer (TNBC. Analysis of cybrids and established breast cancer (BC cell lines showed that metastatic TNBC maintains high levels of ATP through fatty acid β oxidation (FAO and activates Src oncoprotein through autophosphorylation at Y419. Manipulation of FAO including the knocking down of carnitine palmitoyltransferase-1A (CPT1 and 2 (CPT2, the rate-limiting proteins of FAO, and analysis of patient-derived xenograft models confirmed the role of mitochondrial FAO in Src activation and metastasis. Analysis of TCGA and other independent BC clinical data further reaffirmed the role of mitochondrial FAO and CPT genes in Src regulation and their significance in BC metastasis.

  18. Fatty Acid Oxidation-Driven Src Links Mitochondrial Energy Reprogramming and Oncogenic Properties in Triple-Negative Breast Cancer.

    Park, Jun Hyoung; Vithayathil, Sajna; Kumar, Santosh; Sung, Pi-Lin; Dobrolecki, Lacey Elizabeth; Putluri, Vasanta; Bhat, Vadiraja B; Bhowmik, Salil Kumar; Gupta, Vineet; Arora, Kavisha; Wu, Danli; Tsouko, Efrosini; Zhang, Yiqun; Maity, Suman; Donti, Taraka R; Graham, Brett H; Frigo, Daniel E; Coarfa, Cristian; Yotnda, Patricia; Putluri, Nagireddy; Sreekumar, Arun; Lewis, Michael T; Creighton, Chad J; Wong, Lee-Jun C; Kaipparettu, Benny Abraham


    Transmitochondrial cybrids and multiple OMICs approaches were used to understand mitochondrial reprogramming and mitochondria-regulated cancer pathways in triple-negative breast cancer (TNBC). Analysis of cybrids and established breast cancer (BC) cell lines showed that metastatic TNBC maintains high levels of ATP through fatty acid β oxidation (FAO) and activates Src oncoprotein through autophosphorylation at Y419. Manipulation of FAO including the knocking down of carnitine palmitoyltransferase-1A (CPT1) and 2 (CPT2), the rate-limiting proteins of FAO, and analysis of patient-derived xenograft models confirmed the role of mitochondrial FAO in Src activation and metastasis. Analysis of TCGA and other independent BC clinical data further reaffirmed the role of mitochondrial FAO and CPT genes in Src regulation and their significance in BC metastasis.


    Coate, Thomas M.; Swanson, Tracy L.; Copenhaver, Philip F.


    Reverse signaling via GPI-linked Ephrins may help control cell proliferation and outgrowth within the nervous system, but the mechanisms underlying this process remain poorly understood. In the embryonic enteric nervous system (ENS) of the moth Manduca sexta, migratory neurons forming the enteric plexus (EP cells) express a single Ephrin ligand (GPI-linked MsEphrin), while adjacent midline cells that are inhibitory to migration express the cognate receptor (MsEph). Knocking down MsEph receptor expression in cultured embryos with antisense morpholino oligonucleotides allowed the EP cells to cross the midline inappropriately, consistent with the model that reverse signaling via MsEphrin mediates a repulsive response in the ENS. Src family kinases have been implicated in reverse signaling by type-A Ephrins in other contexts, and MsEphrin colocalizes with activated forms of endogenous Src in the leading processes of the EP cells. Pharmacological inhibition of Src within the developing ENS induced aberrant midline crossovers, similar to the effect of blocking MsEphrin reverse signaling. Hyperstimulating MsEphrin reverse signaling with MsEph-Fc fusion proteins induced the rapid activation of endogenous Src specifically within the EP cells, as assayed by Western blots of single embryonic gut explants and by whole-mount immunostaining of cultured embryos. In longer cultures, treatment with MsEph-Fc caused a global inhibition of EP cell migration and outgrowth, an effect that was prevented by inhibiting Src activation. These results support the model that MsEphrin reverse signaling induces the Src-dependent retraction of EP cell processes away from the enteric midline, thereby helping to confine the neurons to their appropriate pathways. PMID:19295147

  20. Liver-specific expressions of HBx and src in the p53 mutant trigger hepatocarcinogenesis in zebrafish.

    Jeng-Wei Lu

    Full Text Available Hepatocarcinogenesis is a multistep process that starts from fatty liver and transitions to fibrosis and, finally, into cancer. Many etiological factors, including hepatitis B virus X antigen (HBx and p53 mutations, have been implicated in hepatocarcinogenesis. However, potential synergistic effects between these two factors and the underlying mechanisms by which they promote hepatocarcinogenesis are still unclear. In this report, we show that the synergistic action of HBx and p53 mutation triggers progressive hepatocellular carcinoma (HCC formation via src activation in zebrafish. Liver-specific expression of HBx in wild-type zebrafish caused steatosis, fibrosis and glycogen accumulation. However, the induction of tumorigenesis by HBx was only observed in p53 mutant fish and occurred in association with the up-regulation and activation of the src tyrosine kinase pathway. Furthermore, the overexpression of src in p53 mutant zebrafish also caused hyperplasia, HCC, and sarcomatoid HCC, which were accompanied by increased levels of the signaling proteins p-erk, p-akt, myc, jnk1 and vegf. Increased expression levels of lipogenic factors and the genes involved in lipid metabolism and glycogen storage were detected during the early stages of hepatocarcinogenesis in the HBx and src transgenic zebrafish. The up-regulation of genes involved in cell cycle regulation, tumor progression and other molecular hallmarks of human liver cancer were found at later stages in both HBx and src transgenic, p53 mutant zebrafish. Together, our study demonstrates that HBx and src overexpression induced hepatocarcinogenesis in p53 mutant zebrafish. This phenomenon mimics human HCC formation and provides potential in vivo platforms for drug screening for therapies for human liver cancer.

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

    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: [Zhejiang University School of Medicine, Hangzhou (China); Xiujun, Cai, E-mail: [Sir Run Run Shaw Hospital of Zhejiang University School of Medicine, Hangzhou (China)


    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.

  2. Identifying redundancy and exposing provenance in crowdsourced data analysis.

    Willett, Wesley; Ginosar, Shiry; Steinitz, Avital; Hartmann, Björn; Agrawala, Maneesh


    We present a system that lets analysts use paid crowd workers to explore data sets and helps analysts interactively examine and build upon workers' insights. We take advantage of the fact that, for many types of data, independent crowd workers can readily perform basic analysis tasks like examining views and generating explanations for trends and patterns. However, workers operating in parallel can often generate redundant explanations. Moreover, because workers have different competencies and domain knowledge, some responses are likely to be more plausible than others. To efficiently utilize the crowd's work, analysts must be able to quickly identify and consolidate redundant responses and determine which explanations are the most plausible. In this paper, we demonstrate several crowd-assisted techniques to help analysts make better use of crowdsourced explanations: (1) We explore crowd-assisted strategies that utilize multiple workers to detect redundant explanations. We introduce color clustering with representative selection--a strategy in which multiple workers cluster explanations and we automatically select the most-representative result--and show that it generates clusterings that are as good as those produced by experts. (2) We capture explanation provenance by introducing highlighting tasks and capturing workers' browsing behavior via an embedded web browser, and refine that provenance information via source-review tasks. We expose this information in an explanation-management interface that allows analysts to interactively filter and sort responses, select the most plausible explanations, and decide which to explore further.

  3. Lapis lazuli provenance study by means of micro-PIXE

    Re, Alessandro, E-mail: [Dipartimento di Fisica Sperimentale, Universita di Torino, Via Giuria 1, 10125 Torino (Italy); INFN Sezione di Torino and Centre of Excellence ' Nanostructured Interfaces and Surfaces' , Universita di Torino, Via P. Giuria 1, 10125 Torino (Italy); Giudice, Alessandro Lo [Dipartimento di Fisica Sperimentale, Universita di Torino, Via Giuria 1, 10125 Torino (Italy); INFN Sezione di Torino and Centre of Excellence ' Nanostructured Interfaces and Surfaces' , Universita di Torino, Via P. Giuria 1, 10125 Torino (Italy); Angelici, Debora [Dipartimento di Fisica Sperimentale, Universita di Torino, Via Giuria 1, 10125 Torino (Italy); Calusi, Silvia; Giuntini, Lorenzo; Massi, Mirko [Dipartimento di Fisica, Universita and INFN Sezione di Firenze, Via Sansone 1, 50019 Sesto Fiorentino, Firenze (Italy); Pratesi, Giovanni [Dipartimento di Scienze della Terra and Museo di Storia Naturale, Universita di Firenze, Via G. La Pira 4, 50121 Firenze (Italy)


    In this paper we report about the micro-PIXE characterisation of lapis lazuli, for a provenance study of this semi-precious stone, used for glyptic as early as 7000 years ago. The final aim is to find markers permitting to identify the origin of the raw material coming from three quarries in regions of historical importance: Afghanistan, Pamir Mountains and Siberia. This may help to reconstruct trade routes, especially for ancient objects for which written testimonies are scanty or absent at all. Due to the heterogeneity of lapis lazuli we concentrate our attention on single phases instead of the whole stone; in particular we focused on two of the main phases: lazurite, responsible for the blue colour, and diopside, the most frequent accessory mineral. This study was preceded and completed by means of microanalysis with Scanning Electron Microscopy (SEM-EDX) and Cold-Cathodoluminescence (cold-CL) analysis. Despite the limited number of analysed samples, results are sufficient to exclude/suggest a few features as provenance markers, partly confirming what has been previously published in literature.

  4. Provenance of Neoproterozoic sedimentary basement of northern Iran, Kahar Formation

    Etemad-Saeed, Najmeh; Hosseini-Barzi, Mahboubeh; Adabi, Mohammad Hossein; Sadeghi, Abbas; Houshmandzadeh, Abdolrahim


    This article presents new data to understand the nature of the hidden crystalline basement of northern Iran and the tectonic setting of Iran during late Neoproterozoic time. The siliciclastic-dominated Kahar Formation represents the oldest known exposures of northern Iran and comprises late Ediacaran (ca. 560-550 Ma) compositionally immature sediments including mudrocks, sandstones, and conglomerates. This work focuses on provenance of three well preserved outcrops of this formation in Alborz Mountains: Kahar Mountain, Sarbandan, and Chalus Road, through petrographic and geochemical methods. Mineralogical Index of Alteration (MIA) and Chemical Index of Alteration (CIA-after correction for K-metasomatism) values combined with A-CN-K relations suggest moderate weathering in the source areas. The polymictic nature of Kahar conglomerates indicates a mixed provenance for them. However, modal analysis of Kahar sandstones (volcanic to plagioclase-rich lithic arkose) and whole rock geochemistry of mudrocks suggest that they are largely first-cycle sediments and that their sources were remarkably late Ediacaran, intermediate-felsic igneous rocks from proximal arc settings. Tectonic setting discrimination diagrams also indicate a convergent plate margin and continental arc related basin for Kahar sediments. This interpretation is supported by the phyllo-tectic to tectic composition and geochemistry of mudrocks. These results reveal the presence of a felsic/intermediate subduction-related basement (∼600-550 Ma) in this region, which provides new constraints on subduction scenario during this time interval in Iran, as a part of the Peri-Gondwanan terranes.

  5. Multivariate analysis in provenance studies: Cerrillos obsidians case, Peru

    Bustamante, A.; Delgado, M.; Latini, R. M.; Bellido, A. V. B.


    We present the preliminary results of a provenance study of obsidians samples from Cerrillos (ca. 800 100 b.c.) using Mössbauer Spectroscopy. The Cerrillos archaeological site, located in the Upper Ica Valley, Peru, is the only Paracas ceremonial center excavated so far. The archaeological data collected suggest the existence of a complex social and economic organization on the south coast of Peru. Provenance research of obsidian provides valuable information about the selection of lithic resources by our ancestors and eventually about the existence of communication routes and exchange networks. We characterized 18 obsidian artifacts samples by Mössbauer spectroscopy from Cerrillos. The spectra, recorded at room temperature using different velocities, are mainly composed of broad asymmetric doublets due to the superposition of at least two quadrupole doublets corresponding to Fe2+ in two different sites (species A and B), one weak Fe3+ doublet (specie C) and magnetic components associated to the presence of small particles of magnetite. Multivariate statistical analysis of the Mössbauer data (hyperfine parameters) allows to defined two main groups of obsidians, reflecting different geographical origins.

  6. Multivariate analysis in provenance studies: Cerrillos obsidians case, Peru

    Bustamante, A., E-mail: [Universidad Nacional Mayor de San Marcos, Facultad de Ciencias Fisicas (Peru); Delgado, M. [Qallta (Peru); Latini, R. M.; Bellido, A. V. B. [UFF, Instituto de Quimica, Depto. Fisico-Quimica (Brazil)


    We present the preliminary results of a provenance study of obsidians samples from Cerrillos (ca. 800-100 b.c.) using Moessbauer Spectroscopy. The Cerrillos archaeological site, located in the Upper Ica Valley, Peru, is the only Paracas ceremonial center excavated so far. The archaeological data collected suggest the existence of a complex social and economic organization on the south coast of Peru. Provenance research of obsidian provides valuable information about the selection of lithic resources by our ancestors and eventually about the existence of communication routes and exchange networks. We characterized 18 obsidian artifacts samples by Moessbauer spectroscopy from Cerrillos. The spectra, recorded at room temperature using different velocities, are mainly composed of broad asymmetric doublets due to the superposition of at least two quadrupole doublets corresponding to Fe{sup 2+} in two different sites (species A and B), one weak Fe{sup 3+} doublet (specie C) and magnetic components associated to the presence of small particles of magnetite. Multivariate statistical analysis of the Moessbauer data (hyperfine parameters) allows to defined two main groups of obsidians, reflecting different geographical origins.

  7. Lapis lazuli provenance study by means of micro-PIXE

    Re, Alessandro; Giudice, Alessandro Lo; Angelici, Debora; Calusi, Silvia; Giuntini, Lorenzo; Massi, Mirko; Pratesi, Giovanni


    In this paper we report about the micro-PIXE characterisation of lapis lazuli, for a provenance study of this semi-precious stone, used for glyptic as early as 7000 years ago. The final aim is to find markers permitting to identify the origin of the raw material coming from three quarries in regions of historical importance: Afghanistan, Pamir Mountains and Siberia. This may help to reconstruct trade routes, especially for ancient objects for which written testimonies are scanty or absent at all. Due to the heterogeneity of lapis lazuli we concentrate our attention on single phases instead of the whole stone; in particular we focused on two of the main phases: lazurite, responsible for the blue colour, and diopside, the most frequent accessory mineral. This study was preceded and completed by means of microanalysis with Scanning Electron Microscopy (SEM-EDX) and Cold-Cathodoluminescence (cold-CL) analysis. Despite the limited number of analysed samples, results are sufficient to exclude/suggest a few features as provenance markers, partly confirming what has been previously published in literature.

  8. Clinical differences among PCR-proven dengue serotype infections.

    Limkittikul, Kriengsak; Yingsakmongkon, Sangchai; Jittmittraphap, Akanitt; Chuananon, Somchai; Kongphrai, Yuphin; Kowasupathr, Surasak; Rojanawatsirivit, Chaiyaporn; Mammen, Mammen P; Jampangern, Wipawee


    The objective of this study was to compare the clinical spectra of the dengue serotypes proven by the PCR technique. This retrospective study reviewed the clinical information of dengue-infected patients who were admitted to northeastern provincial hospitals in Thailand from June to September 2002. Dengue infection and viral serotypes were confirmed by polymerase chain reaction (PCR). Paired anti-dengue immunoglobulin G (IgG) and IgM from paired sera were analyzed by enzyme-linked immunosorbent assay (ELISA). Ninety-nine PCR-proven dengue-infected Thai patients were studied. Their ages ranged from 3-30 years. They were infected with DEN1, DEN2, DEN3 and DEN4 in 21, 55, 12, and 12%, respectively. Twenty-two percent had primary and 78% had secondary infections. Dengue fever was the most common presentation for both primary (77.2%) and secondary infections (46.7%). The ratios of dengue fever:dengue hemorrhagic fever (DF:DHF) and non-dengue shock syndrome:dengue shock syndrome (non-DSS:DSS) for DEN2 was the lowest of the dengue serotypes. There was no difference in the duration of fever, percentage of hepatomegaly and bleeding among the serotypes in both DF and DHF. The trends in the white blood cells, lymphocyte and atypical lymphocyte counts in DEN3 were the highest, while those of DEN1 were the lowest of the dengue serotypes.

  9. Preventing aggressive prostate cancer with proven cardiovascular disease preventive methods

    Mark A Moyad


    Full Text Available Cardiovascular disease (CVD has been the number one cause of death in the U.S. for 114 of the last 115 years. Risk factors for prostate cancer have primarily mirrored risk proven risk factors for CVD, especially aggressive disease. Obesity, dyslipidemia, glucose intolerance, metabolic syndrome, unhealthy dietary habits or caloric excess, lack of physical activity, and inflammation are just some of these shared risk factors. The evidence also suggests proven CVD preventive measures are identical to prostate cancer preventive measures, especially in regard to aggressive disease. Thus, apart from lifestyle measures that can encourage optimal heart and prostate health there are potentially several dietary supplements that need to be avoided in healthy men because they may also increase the risk of prostate cancer. However, there are also several low-cost, generic, safe in the appropriate individuals, and naturally derived agents that could reduce prostate cancer risk, and these can be discussed and remembered utilizing the acronym S.A.M. (statins, aspirin, and/or metformin.

  10. MRI of pathology-proven peripheral nerve amyloidosis

    McKenzie, Gavin A.; Broski, Stephen M.; Howe, Benjamin M.; Spinner, Robert J.; Amrami, Kimberly K.; Dispenzieri, Angela; Ringler, Michael D. [Mayo Clinic, Department of Musculoskeletal Radiology, Rochester, MN (United States)


    To highlight the MRI characteristics of pathologically proven amyloidosis involving the peripheral nervous system (PNS) and determine the utility of MRI in directing targeted biopsy for aiding diagnosis. A retrospective study was performed for patients with pathologically proven PNS amyloidosis who also underwent MRI of the biopsied or excised nerve. MRI signal characteristics, nerve morphology, associated muscular denervation changes, and the presence of multifocal involvement were detailed. Pathology reports were reviewed to determine subtypes of amyloid. Charts were reviewed to gather patient demographics, neurological symptoms and radiologist interpretation. Four men and three women with a mean age of 62 ± 11 years (range 46-76) were identified. All patients had abnormal findings on EMG with mixed sensorimotor neuropathy. All lesions demonstrated diffuse multifocal neural involvement with T1 hypointensity, T2 hyperintensity, and variable enhancement on MRI. One lesion exhibited superimposed T2 hypointensity. Six of seven patients demonstrated associated muscular denervation changes. Peripheral nerve amyloidosis is rare, and the diagnosis is difficult because of insidious symptom onset, mixed sensorimotor neurologic deficits, and the potential for a wide variety of nerves affected. On MRI, peripheral nerve involvement is most commonly characterized by T1 hypointensity, T2 hyperintensity, variable enhancement, maintenance of the fascicular architecture with fusiform enlargement, multifocal involvement and muscular denervation changes. While this appearance mimics other inflammatory neuropathies, MRI can readily detect neural changes and direct-targeted biopsy, thus facilitating early diagnosis and appropriate management. (orig.)

  11. Differential regulation of T cell antigen responsiveness by isoforms of the src-related tyrosine protein kinase p59fyn


    Recent observations suggest that the src-related tyrosine protein kinase p59fyn may be involved in antigen-induced T lymphocyte activation. As a result of alternative splicing, p59fyn exists as two isoforms that differ exclusively within a short sequence spanning the end of the Src Homology 2 (SH2) region and the beginning of the tyrosine protein kinase domain. While one p59fyn isoform (fynB) is highly expressed in brain, the alternative product (fynT) is principally found in T lymphocytes. T...

  12. Ontology-Driven Provenance Management in eScience: An Application in Parasite Research

    Sahoo, Satya S.; Weatherly, D. Brent; Mutharaju, Raghava; Anantharam, Pramod; Sheth, Amit; Tarleton, Rick L.

    Provenance, from the French word "provenir", describes the lineage or history of a data entity. Provenance is critical information in scientific applications to verify experiment process, validate data quality and associate trust values with scientific results. Current industrial scale eScience projects require an end-to-end provenance management infrastructure. This infrastructure needs to be underpinned by formal semantics to enable analysis of large scale provenance information by software applications. Further, effective analysis of provenance information requires well-defined query mechanisms to support complex queries over large datasets. This paper introduces an ontology-driven provenance management infrastructure for biology experiment data, as part of the Semantic Problem Solving Environment (SPSE) for Trypanosoma cruzi (T.cruzi). This provenance infrastructure, called T.cruzi Provenance Management System (PMS), is underpinned by (a) a domain-specific provenance ontology called Parasite Experiment ontology, (b) specialized query operators for provenance analysis, and (c) a provenance query engine. The query engine uses a novel optimization technique based on materialized views called materialized provenance views (MPV) to scale with increasing data size and query complexity. This comprehensive ontology-driven provenance infrastructure not only allows effective tracking and management of ongoing experiments in the Tarleton Research Group at the Center for Tropical and Emerging Global Diseases (CTEGD), but also enables researchers to retrieve the complete provenance information of scientific results for publication in literature.

  13. Haemophilus ducreyi targets Src family protein tyrosine kinases to inhibit phagocytic signaling.

    Mock, Jason R; Vakevainen, Merja; Deng, Kaiping; Latimer, Jo L; Young, Jennifer A; van Oers, Nicolai S C; Greenberg, Steven; Hansen, Eric J


    Haemophilus ducreyi, the etiologic agent of the sexually transmitted disease chancroid, has been shown to inhibit phagocytosis of both itself and secondary targets in vitro. Immunodepletion of LspA proteins from H. ducreyi culture supernatant fluid abolished this inhibitory effect, indicating that the LspA proteins are necessary for the inhibition of phagocytosis by H. ducreyi. Fluorescence microscopy revealed that macrophages incubated with wild-type H. ducreyi, but not with a lspA1 lspA2 mutant, were unable to complete development of the phagocytic cup around immunoglobulin G-opsonized targets. Examination of the phosphotyrosine protein profiles of these two sets of macrophages showed that those incubated with wild-type H. ducreyi had greatly reduced phosphorylation levels of proteins in the 50-to-60-kDa range. Subsequent experiments revealed reductions in the catalytic activities of both Lyn and Hck, two members of the Src family of protein tyrosine kinases that are known to be involved in the proximal signaling steps of Fcgamma receptor-mediated phagocytosis. Additional experiments confirmed reductions in the levels of both active Lyn and active Hck in three different immune cell lines, but not in HeLa cells, exposed to wild-type H. ducreyi. This is the first example of a bacterial pathogen that suppresses Src family protein tyrosine kinase activity to subvert phagocytic signaling in hostcells.

  14. The Csk-binding protein PAG regulates PDGF-induced Src mitogenic signaling via GM1

    Veracini, Laurence; Simon, Valérie; Richard, Véronique; Schraven, Burkhart; Horejsi, Vaclav; Roche, Serge; Benistant, Christine


    Spatial regulation is an important feature of signal specificity elicited by cytoplasmic tyrosine kinases of the Src family (SRC family protein tyrosine kinases [SFK]). Cholesterol-enriched membrane domains, such as caveolae, regulate association of SFK with the platelet-derived growth factor receptor (PDGFR), which is needed for kinase activation and mitogenic signaling. PAG, a ubiquitously expressed member of the transmembrane adaptor protein family, is known to negatively regulate SFK signaling though binding to Csk. We report that PAG modulates PDGFR levels in caveolae and SFK mitogenic signaling through a Csk-independent mechanism. Regulation of SFK mitogenic activity by PAG requires the first N-terminal 97 aa (PAG-N), which include the extracellular and transmembrane domains, palmitoylation sites, and a short cytoplasmic sequence. We also show that PAG-N increases ganglioside GM1 levels at the cell surface and, thus, displaces PDGFR from caveolae, a process that requires the ganglioside-specific sialidase Neu-3. In conclusion, PAG regulates PDGFR membrane partitioning and SFK mitogenic signaling by modulating GM1 levels within caveolae independently from Csk. PMID:18695048

  15. MICA Expression Is Regulated by Cell Adhesion and Contact in a FAK/Src-Dependent Manner

    Moncayo, Gerald; Lin, Da; McCarthy, Michael T.; Watson, Aleksandra A.; O’Callaghan, Christopher A.


    MICA is a major ligand for the NKG2D immune receptor, which plays a key role in activating natural killer (NK) cells and cytotoxic T cells. We analyzed NKG2D ligand expression on a range of cell types and could demonstrate that MICA expression levels were closely linked to cellular growth mode. While the expression of other NKG2D ligands was largely independent of cell growth mode, MICA expression was mainly found on cells cultured as adherent cells. In addition, MICA surface expression was reduced through increase in cell–cell contact or loss of cell–matrix adherence. Furthermore, we found that the reduction in MICA expression was modulated by focal adhesion kinase (FAK)/Src signaling and associated with increased susceptibility to NK cell-mediated killing. While the mechanisms of tumor immune evasion are not fully understood, the reduction of MICA expression following loss of attachment poises a potential way by which metastasizing tumor cells avoid immune detection. The role of FAK/Src in this process indicates a potential therapeutic approach to modulate MICA expression and immune recognition of tumor cells during metastasis. PMID:28154561

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

    Jueun Oh


    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.

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


    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.

  18. Baicalin and Baicalein Inhibit Src Tyrosine Kinase and Production of IL-6

    Dubravko Jelić


    Full Text Available Flavonoids play an important role in the treatment of various diseases, as they are able to inhibit reactive oxygen species, which cause damage to cells and tissues which may lead to increased risk of inflammatory diseases. Baicalin and baicalein, two flavonoids found in the roots of Scutellaria baicalensis, in the leaves of Thymus vulgaris and Oroxylum indicum, were tested for their anti-inflammatory activity as well as for their cytotoxicity. Thereby the two compounds were investigated on Src tyrosine kinase inhibition and inhibition of production of interleukin (IL-6 in lipopolysaccharide- (LPS- stimulated THP-1 cells. Additionally, the THP-1 cell line was used for the determination of the cytotoxicity. Both baicalin and baicalein showed some anti-inflammatory properties, while baicalein turned out to be the more active compound with higher inhibitory activities on both Src tyrosine kinase and production of cytokine IL-6. Baicalin and baicalein showed no signs of cytotoxicity in the MTS cytotoxicity assay in THP-1 cells.

  19. BMP2 rescues deficient cell migration in Tgfbr3(-/-) epicardial cells and requires Src kinase.

    Allison, Patrick; Espiritu, Daniella; Camenisch, Todd D


    During embryogenesis, the epicardium undergoes proliferation, migration, and differentiation into several cardiac cell types which contribute to the coronary vessels. The type III transforming growth factor-β receptor (TGFβR3) is required for epicardial cell invasion and development of coronary vasculature in vivo. Bone Morphogenic Protein-2 (BMP2) is a driver of epicardial cell migration. Utilizing a primary epicardial cell line derived from Tgfbr3(+/+) and Tgfbr3(-/-) mouse embryos, we show that Tgfbr3(-/-) epicardial cells are deficient in BMP2 mRNA expression. Tgfbr3(-/-) epicardial cells are deficient in 2-dimensional migration relative to Tgfbr3(+/+) cells; BMP2 induces cellular migration to Tgfbr3(+/+) levels without affecting proliferation. We further demonstrate that Src kinase activity is required for BMP2 driven Tgfbr3(-/-) migration. BMP2 also requires Src for filamentous actin polymerization in Tgfbr3(-/-) epicardial cells. Taken together, our data identifies a novel pathway in epicardial cell migration required for development of the coronary vessels.

  20. Solvent Refined Coal (SRC) process. [Runs 49 to 57 and 59 to 62



    This report summarizes the progress of the Solvent Refined Coal (SRC) project for the period January 1, 1979 through December 31, 1979. The fourth quarter of 1979 is reported here in detail. Turnaround activities at the Fort Lewis SRC-II Pilot Plant were completed. During the shutdown, installation of Slurry Preheater B was completed. In addition, extensive modifications were completed to improve operability and slurry handling capabilities. The experimental program for testing Slurry Preheater B was revised to improve the data base for design scale-up considerations. Coal feed was established using Powhatan No. 6 coal. Twenty slurry survey tests were designed to establish the effects of varous slurry and heater inlet hydrogen flow rates on heat transfer, heater coil pressure drop, and heater operability. Additional tests were also added to the preheater evaluation program to study the effects of coal concentration, recycle pyridine insoluble concentration and preheater outlet temperatures. During 1979, PDU P-99 completed 13 runs (Runs 49 to 57 and 59 to 62). All these runs were made feeding coal from the Powhatan No. 5 Mine.

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

    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.

  2. Structural and Biochemical Basis for Intracellular Kinase Inhibition by Src-specific Peptidic Macrocycles.

    Aleem, Saadat; Georghiou, George; Kleiner, Ralph E; Guja, Kip; Craddock, Barbara P; Lyczek, Agatha; Chan, Alix I; Garcia-Diaz, Miguel; Miller, W Todd; Liu, David R; Seeliger, Markus A


    Protein kinases are attractive therapeutic targets because their dysregulation underlies many diseases, including cancer. The high conservation of the kinase domain and the evolution of drug resistance, however, pose major challenges to the development of specific kinase inhibitors. We recently discovered selective Src kinase inhibitors from a DNA-templated macrocycle library. Here, we reveal the structural basis for how these inhibitors retain activity against a disease-relevant, drug-resistant kinase mutant, while maintaining Src specificity. We find that these macrocycles display a degree of modularity: two of their three variable groups interact with sites on the kinase that confer selectivity, while the third group interacts with the universally conserved catalytic lysine and thereby retains the ability to inhibit the "gatekeeper" kinase mutant. We also show that these macrocycles inhibit migration of MDA-MB-231 breast tumor cells. Our findings establish intracellular kinase inhibition by peptidic macrocycles, and inform the development of potent and specific kinase inhibitors. Copyright © 2016 Elsevier Ltd. All rights reserved.

  3. Src/Syk-Targeted Anti-Inflammatory Actions of Triterpenoidal Saponins from Gac (Momordica cochinchinensis) Seeds.

    Yu, Jae Sik; Kim, Jun Ho; Lee, Seulah; Jung, Kiwon; Kim, Ki Hyun; Cho, Jae Youl


    Momordica cochinchinensis Spreng (family Cucurbitaceae), also known as gac, or red melon, is an edible Southeast Asian fruit valued for its nutritional and medicinal properties. Specifically, Momordicae Semen, the seeds of the gac fruit, is used in traditional Chinese medicine to treat boils, rheumatic pain, muscle spasm, hemorrhoids, and hemangiomas. In this study, a chemical investigation into a gac seed ethanol (EtOH) extract resulted in the identification of three triterpenoidal saponins (1-3), which were investigated for their anti-inflammatory effects. Among the saponins, momordica saponin I (compound 3) reduced the production of nitric oxide (NO) in LPS-activated RAW264.7 cells without inducing cytotoxicity. The mRNA levels of inducible NO synthase (iNOS) and cyclooxygenase (COX)-2 were decreased by momordica saponin I. Additionally, the translocation of p65 and p50 (subunits of the transcription factor NF-[Formula: see text]B) into the nucleus was remarkably inhibited. Furthermore, the phosphorylation levels of inflammatory signaling proteins (I[Formula: see text]B[Formula: see text], Src, and Syk) known to be upstream regulatory molecules of p65 were decreased under momordica saponin I-treated conditions. The molecular targets of momordica saponin I were confirmed in overexpression experiments and through immunoblot analyses with Src and Syk. This study provides evidence that momordica saponin I could be beneficial in treating inflammatory diseases, and should be considered a bioactive immunomodulatory agent with anti-inflammatory properties.

  4. Geochemistry and provenance of the Carboniferous Baixo Alentejo Flysch Group, South Portuguese Zone

    Jorge, R. C. G. S.; Fernandes, P.; Rodrigues, B.; Pereira, Z.; Oliveira, J. T.


    This work is focused on the turbiditic sediments from the Carboniferous Baixo Alentejo Flysch Group (BAFG) in the South Portuguese Zone, an external zone of the Iberian Variscides. The aim of this work is to constrain the provenance and tectonic setting of these sediments in a context of a complex evolution of SW Iberian Variscides. For this purpose, we performed a systematic study of petrographical and geochemical signatures of greywackes and shales from the three BAFG formations: Mértola, Mira and Brejeira. Major and trace element composition and ratios suggest heterogeneous source area composition for BAFG shales and greywackes. For the oldest Mértola Formation greywackes, source area is dominated by granitoid rocks with minor mafic input. The latter becomes residual in the Mira Formation. The youngest Brejeira Formation greywackes show clear felsic affiliation associated with an increase in recycled components. The shales of all three BAFG formations denote a granodioritic affiliation. Chemical Index of Alteration (CIA) and Plagioclase Index of Alteration (PIA) values suggest moderate weathering in the source areas of Mértola and Mira formations. These indices, together with A-CN-K relations, point out to steady-state weathering conditions in the source areas of both formations. In contrast, both CIA and PIA values for the Brejeira Formation indicate variable conditions of palaeoweathering, from moderate to intense, as a consequence of non-steady-state conditions probably triggered by tectonic instability in the provenance area. Compared to the greywackes, the shales of all three BAFG formations exhibit higher CIA and PIA values, as well as low K2O/Al2O3 (~ 0.2) and index of compositional variability (Morena Zone (Gondwanan affinity), with possible contribution from an external (Avalonian) source.

  5. Sports drug testing: Analytical aspects of selected cases of suspected, purported, and proven urine manipulation.

    Thevis, Mario; Geyer, Hans; Sigmund, Gerd; Schänzer, Wilhelm


    Manipulation of urine specimens provided by elite athletes for doping control purposes has been reported several times in the past, and in most of these cases urine substitution was eventually proven. Recent findings of suspected and substantiated manipulation have outlined the complexity and diversity of tampering options, sample appearance alterations resulting from non-manipulative influence, and the analytical challenges arising from these scenarios. Using state-of-the-art mass spectrometric and immunological doping control and forensic chemistry methodologies, four unusual findings were observed. One sports drug testing specimen was found to contain an unusually high content of saccharides accompanied by hordenine and Serpine-Z4, while no endogenous steroid (e.g. testosterone, epitestosterone, androsterone and etiocholanolone) was detected. This specimen was identified as non-alcoholic beer filled into the doping control sample container, constituting an undisputed doping offense. A doping control sample of bright green color was received and found to contain residues of methylene blue, which is not considered relevant for doping controls as no masking or manipulative effect is known. In addition, the number of urine samples of raspberry to crimson red coloration received at doping control laboratories has constantly increased during the last years, attributed to the presence of hemoglobin or betanin/isobetanin. Also here, no doping rule violation was given and an impact on routine analytical results was not observed. Finally, a total of 8 sports drug testing samples collected at different competition sites was shown to contain identical urine specimens as indicated by steroid profile analysis and conclusively proven by DNA-STR (short tandem repeat) analysis. Here, the athletes in question were not involved in the urine substitution act but the doping control officer was convicted of sample manipulation.

  6. Expression of a phosphorylated substrate domain of p130Cas promotes PyMT-induced c-Src-dependent murine breast cancer progression.

    Zhao, Yingshe; Kumbrink, Joerg; Lin, Bor-Tyh; Bouton, Amy H; Yang, Shi; Toselli, Paul A; Kirsch, Kathrin H


    Elevated expression of p130Cas (Crk-associated substrate)/BCAR1 (breast cancer antiestrogen resistance 1) in human breast tumors is a marker of poor prognosis and poor overall survival. p130Cas is a downstream target of the tyrosine kinase c-Src. Signaling mediated by p130Cas through its phosphorylated substrate domain (SD) and interaction with effector molecules directly promotes tumor progression. We previously developed a constitutively phosphorylated p130Cas SD molecule, Src*/SD (formerly referred to as Src*/CasSD), which acts as decoy molecule and attenuates the transformed phenotype in v-crk-transformed murine fibroblasts and human breast cancer cells. To test the function of this molecule in vivo, we established mouse mammary tumor virus (MMTV)-long terminal repeat-Src*/SD transgenic mice in which mammary gland development and tumor formation were analyzed. Transgenic expression of the Src*/SD molecule under the MMTV-long terminal repeat promoter did not interfere with normal mammary gland development or induce tumors in mice observed for up to 11 months. To evaluate the effects of the Src*/SD molecule on tumor development in vivo, we utilized the MMTV-polyoma middle T-antigen (PyMT) murine breast cancer model that depends on c-Src. PyMT mice crossed with Src*/SD mice displayed accelerated tumor formation. The earlier onset of tumors can be explained by the interaction of the Src* domain with PyMT and targeting the fused phosphorylated SD to the membrane. At membrane compartments, it might integrate membrane-associated active signaling complexes leading to increased proliferation measured by phospho-Histone H3 staining. Although these results were unexpected, they emphasize the importance of preventing the membrane association of Src*/SD when employed as decoy molecule.

  7. Can cathodoluminescence of feldspar be used as provenance indicator?

    Scholonek, Christiane; Augustsson, Carita


    We have studied feldspar from crystalline rocks for its textural and spectral cathodoluminescence (CL) characteristics with the aim to reveal their provenance potential. We analyzed ca. 60 rock samples of plutonic, volcanic, metamorphic, and pegmatitic origin from different continents and of 16 Ma to 2 Ga age for their feldspar CL textures and ca. 1200 feldspar crystals from these rocks for their CL color spectra. Among the analyzed rocks, igneous feldspar is most commonly zoned, whereby oscillatory zoning can be confirmed to be typical for volcanic plagioclase. The volcanic plagioclase also less commonly contains twin lamellae that are visible in CL light than crystals from other rock types. Alkali feldspar, particularly from igneous and pegmatitic rocks, was noted to be most affected by alteration features, visible as dark spots, lines and irregular areas. The size of all textural features of up to ca. 150 μm, in combination with possible alteration in both the source area and the sedimentary system, makes the CL textures of feldspar possible to use for qualitative provenance research only. We observed alkali feldspar mostly to luminesce in a bluish color and sometimes in red, and plagioclase in green to yellow. The corresponding CL spectra are dominated by three apparent intensity peaks at 440-520 nm (mainly blue), 540-620 nm (mainly green) and 680-740 nm (red to infrared). A dominance of the peak in the green wavelength interval over the blue one for plagioclase makes CL particularly useful for the differentiation of plagioclase from alkali feldspar. An apparent peak position in red to infrared at < 710 nm for plagioclase mainly is present in mafic rocks. Present-day coastal sand from Peru containing feldspar with the red to infrared peak position mainly exceeding 725 nm for northern Peruvian sand and a larger variety for sand from southern Peru illustrates a discriminative effect of different source areas. We conclude that the provenance application

  8. Provenance Capture in Data Access and Data Manipulation Software

    West, P.; Fox, P. A.; McGuinness, D. L.; Gallagher, J. H.; Holloway, D.; Potter, N.


    There is increasing need to trace back the origins of data products, whether images or charts in a report, data obtained from a sensor on an instrument, a generated dataset referenced in a research paper, in government reports on the environment, or in a publication or poster presentation. Yet, most software applications that perform data access and manipulation keep only limited history of the data, i.e. the provenance. Imagine the following scenario: There is a figure in a report showing multiple graphs and plots related to global climate, the report is being drafted for a government agency. The graphs and plots are generated using an algorithm from an iPython Notebook, developed by a researcher who is using a particular data portal, where the algorithm pulls data from four data sets from that portal. That data is aggregated together over the time dimension, constrained to a few parameters, accessed using a particular piece of data access software, and converted from one datatype to another datatype; All the processing on the data sets was conducted by three different researchers from a public university, on a project funded by the same government agency requesting the report, with one Principal Investigator and two Co-Investigators. In this scenario, today we're lucky to get a blob of text under the figure that might say a couple things about the figure with a reference to a publication that was written a few years ago. Data citation, data publishing information, licensing information, and provenance are all lacking in the derived data products. What we really want is to be able to trace the figure all the way back to the original datasets, including what was done to those datasets; and to see information about the researchers, the project, the agency funding, the award, and the organizations collaborating on the project. In this paper we discuss the need for such information and traceback features, as well as new technologies and standards that can help us

  9. Src-Like adaptor protein (SLAP) binds to the receptor tyrosine kinase Flt3 and modulates receptor stability and downstream signaling.

    Kazi, Julhash U; Rönnstrand, Lars


    Fms-like tyrosine kinase 3 (Flt3) is an important growth factor receptor in hematopoiesis. Gain-of-function mutations of the receptor contribute to the transformation of acute myeloid leukemia (AML). Src-like adaptor protein (SLAP) is an interaction partner of the E3 ubiquitin ligase Cbl that can regulate receptor tyrosine kinases-mediated signal transduction. In this study, we analyzed the role of SLAP in signal transduction downstream of the type III receptor tyrosine kinase Flt3. The results show that upon ligand stimulation SLAP stably associates with Flt3 through multiple phosphotyrosine residues in Flt3. SLAP constitutively interacts with oncogenic Flt3-ITD and co-localizes with Flt3 near the cell membrane. This association initiates Cbl-dependent receptor ubiquitination and degradation. Depletion of SLAP expression by shRNA in Flt3-transfected Ba/F3 cells resulted in a weaker activation of FL-induced PI3K-Akt and MAPK signaling. Meta-analysis of microarray data from patient samples suggests that SLAP mRNA is differentially expressed in different cancers and its expression was significantly increased in patients carrying the Flt3-ITD mutation. Thus, our data suggest a novel role of SLAP in different cancers and in modulation of receptor tyrosine kinase signaling apart from its conventional role in regulation of receptor stability.

  10. Src-Like adaptor protein (SLAP binds to the receptor tyrosine kinase Flt3 and modulates receptor stability and downstream signaling.

    Julhash U Kazi

    Full Text Available Fms-like tyrosine kinase 3 (Flt3 is an important growth factor receptor in hematopoiesis. Gain-of-function mutations of the receptor contribute to the transformation of acute myeloid leukemia (AML. Src-like adaptor protein (SLAP is an interaction partner of the E3 ubiquitin ligase Cbl that can regulate receptor tyrosine kinases-mediated signal transduction. In this study, we analyzed the role of SLAP in signal transduction downstream of the type III receptor tyrosine kinase Flt3. The results show that upon ligand stimulation SLAP stably associates with Flt3 through multiple phosphotyrosine residues in Flt3. SLAP constitutively interacts with oncogenic Flt3-ITD and co-localizes with Flt3 near the cell membrane. This association initiates Cbl-dependent receptor ubiquitination and degradation. Depletion of SLAP expression by shRNA in Flt3-transfected Ba/F3 cells resulted in a weaker activation of FL-induced PI3K-Akt and MAPK signaling. Meta-analysis of microarray data from patient samples suggests that SLAP mRNA is differentially expressed in different cancers and its expression was significantly increased in patients carrying the Flt3-ITD mutation. Thus, our data suggest a novel role of SLAP in different cancers and in modulation of receptor tyrosine kinase signaling apart from its conventional role in regulation of receptor stability.

  11. Federated provenance of oceanographic research cruises: from metadata to data

    Thomas, Rob; Leadbetter, Adam; Shepherd, Adam


    The World Wide Web Consortium's Provenance Data Model and associated Semantic Web ontology (PROV-O) have created much interest in the Earth and Space Science Informatics community (Ma et al., 2014). Indeed, PROV-O has recently been posited as an upper ontology for the alignment of various data models (Cox, 2015). Similarly, PROV-O has been used as the building blocks of a data release lifecycle ontology (Leadbetter & Buck, 2015). In this presentation we show that the alignment between different local data descriptions of an oceanographic research cruise can be achieved through alignment with PROV-O and that descriptions of the funding bodies, organisations and researchers involved in a cruise and its associated data release lifecycle can be modelled within a PROV-O based environment. We show that, at a first-order, this approach is scalable by presenting results from three endpoints (the Biological and Chemical Oceanography Data Management Office at Woods Hole Oceanographic Institution, USA; the British Oceanographic Data Centre at the National Oceanography Centre, UK; and the Marine Institute, Ireland). Current advances in ontology engineering, provide pathways to resolving reasoning issues from varying perspectives on implementing PROV-O. This includes the use of the Information Object design pattern where such edge cases as research cruise scheduling efforts are considered. PROV-O describes only things which have happened, but the Information Object design pattern allows for the description of planned research cruises through its statement that the local data description is not the the entity itself (in this case the planned research cruise) and therefore the local data description itself can be described using the PROV-O model. In particular, we present the use of the data lifecycle ontology to show the connection between research cruise activities and their associated datasets, and the publication of those data sets online with Digital Object Identifiers and

  12. Functional Requirements for Information Resource Provenance on the Web

    McCusker, James P.; Lebo, Timothy; Graves, Alvaro; Difranzo, Dominic; Pinheiro da Silva, Paulo; McGuinness, Deborah L.


    We provide a means to formally explain the relationship between HTTP URLs and the representations returned when they are requested. According to existing World Wide Web architecture, the URL serves as an identier for a semiotic referent while the document returned via HTTP serves as a representation of the same referent. This begins with two sides of a semiotic triangle; the third side is the relationship between the URL and the representation received. We complete this description by extending the library science resource model Functional Requirements for Bibliographic Resources (FRBR) with cryptographic message and content digests to create a Functional Requirements for Information Resources (FRIR). We show how applying the FRIR model to HTTP GET and POST transactions disambiguates the many relationships between a given URL and all representations received from its request, provides fine-grained explanations that are complementary to existing explanations of web resources, and integrates easily into the emerging W3C provenance standard.

  13. Optimal management of biopsy-proven low-grade gastricdysplasia

    Jung-Wook Kim; Jae Young Jang


    Gastric adenocarcinoma generally culminates via theinflammation-metaplasia-dysplasia-carcinoma sequenceprogression. The prevalence of gastric adenomasshows marked geographic variation. Recently, therate of diagnosis of low-grade dysplasia (LGD) hasincreased due to increased use of upper endoscopy.Many investigators have reported that gastric highgradedysplasia has high potential for malignancy andshould be removed; however, the treatment for gastricLGD remains controversial. Although the risk of LGDprogression to invasive carcinoma has been reported tobe inconsistent, progression has been observed duringfollow-up. Additionally, the rate of upgraded diagnosisin biopsy-proven LGD is high. Therefore, endoscopicresection (ER) may be useful in the treatment anddiagnosis of LGD, especially if lesions are found to haverisk factors for upgraded histology after ER, such aslarge size, surface erythema or depressed morphology.Fatal complications in endoscopic submucosal dissection(ESD) are extremely low and its therapeutic and diagnosticoutcomes are excellent. Therefore, ESD shouldbe applied preferentially instead of endoscopic mucosalresection.

  14. Using Provenance to support Good Laboratory Practice in Grid Environments

    Ney, Miriam; Schreiber, Andreas


    Conducting experiments and documenting results is daily business of scientists. Good and traceable documentation enables other scientists to confirm procedures and results for increased credibility. Documentation and scientific conduct are regulated and termed as "good laboratory practice." Laboratory notebooks are used to record each step in conducting an experiment and processing data. Originally, these notebooks were paper based. Due to computerised research systems, acquired data became more elaborate, thus increasing the need for electronic notebooks with data storage, computational features and reliable electronic documentation. As a new approach to this, a scientific data management system (DataFinder) is enhanced with features for traceable documentation. Provenance recording is used to meet requirements of traceability, and this information can later be queried for further analysis. DataFinder has further important features for scientific documentation: It employs a heterogeneous and distributed data...

  15. Multi-Method Provenance Analysis of Namibian Desert Sand

    Vermeesch, P.; Garzanti, E.


    Mineralogical, geochemical and geochronological provenance proxies each have their own strengths and weaknesses: a. Bulk geochemistry, framework petrography and heavy mineral compositions can differentiate between source areas characterised by different lithologies, but are sensitive to hydraulic sorting and chemical alteration. b. Detrital zircon U-Pb geochronology is insensitive to winnowing effects, but is 'blind' to lithologies devoid of zircon and cannot differentiate between first cycle and recycled sediments. c. Cosmogenic neon isotopes can be used to identify different generations of surface exposure while simultaneously tracking different magmatic sources. The challenge is then to combine these different proxies into a self consistent story, and do so in as objective a manner as possible. We here present a case study of Namibia's Namib Sand Sea and Skeleton Coast ergs, in which all the aforementioned methods have been combined using a three-way multidimensional scaling (aka INDividual Differences SCALing or INDSCAL) analysis: 1. Each of the datasets was represented by a 'dissimilarity matrix' of pairwise distances between samples. 2. The set of these matrices was fed into the INDSCAL algorithm, which produces two pieces of graphical output: the 'group configuration', which is a scatter plot or 'map' in which similar samples plot close together and dissimilar samples plot far apart, and the 'proxy weights', in which not the samples but the proxies are plotted according to the weight they attached to the 'group configuration' axes. The INDSCAL map of the Namibia dataset indicates that (a) long-shore drift of Orange River sediments dominates the coastal sediment compositions all along the Namibian coast until Angola, and (b) that light and heavy minerals tell complementary parts of the provenance story.

  16. Species choice, provenance and species trials among native Brazilian species

    Drumond, M.A.


    Six papers from the conference are presented. Drumond, M.A., Potential of species native to the semi-arid tropics, 766-781, (Refs. 18), reports on Anadenanthera macrocarpa, Mimosa species, Schinopsis brasiliensis, Spondias tuberosa, Ziziphus joazeiro, Cnidoscolus phyllacanthus, Bursera leptophleos (leptophloeos), Tabebuia impetiginosa, Astronium urundeuva, and Mimosa caesalpinia. Monteiro, R.F.R., Speltz, R.M., Gurgel, J.T. do A.; Silvicultural performance of 24 provenances of Araucaria angustifolia in Parana, 814-824, (Refs. 8). Pires, C.L. da S., Kalil Filho, A.N., Rosa, P.R.F. da, Parente, P.R., Zanatto, A.C.S.; Provenance trials of Cordia alliodora in the State of Sao Paulo, 988-995, (Refs. 9). Nogueira, J.C.B., Siqueira, A.C.M.F., Garrido, M.A.O., Gurgel Garrido, L.M. do A., Rosa, P.R.F., Moraes, J.L. de, Zandarin, M.A., Gurgel Filho, O.A., Trials of some native species in various regions of the State of Sao Paulo, 1051-1063, (Refs. 9) describes Centrolobium tomentosum, Peltophorum dubium, Tabebuia vellosoi, Cariniana legalis, and Balfourodendron riedelianum. Batista, M.P., Borges, J.F., Franco, M.A.B.; Early growth of a native species in comparison with exotics in northeastern Para, Brazil, 1105-1110, (Refs. 3). Jacaranda copaia is compared with Gmelina arborea, Pinus caribaea various hondurensis, Eucalyptus deglupta, and E. urophylla. Lima, P.C.F., Souza, S.M. de, Drumond, M.A.; Trials of native forest species at Petrolina, Pernambuco, 1139-1148, (Refs. 8), deals with Anadenanthera macrocarpa, Piptadenia obliqua, Pithecellobium foliolosum, Astronium urundeuva, Schinopsis brasiliensis, Cassia excelsa, Caesalpinia pyramidalis, Parkia platycephala, Pseudobombax simplicifolium, Tabebuia impetiginosa, Caesalpinia ferrea, and Aspidosperma pyrifolium. 18 references.

  17. Information Diffusion and Provenance in Online Social Media Networks

    Naman Goel


    Full Text Available Traditional data mining techniques are going through an extensive research so as to suit their application in the dynamically evolving social media networks which are not only large in size but also require real time processing of data streams. One such application which is of social, industrial and political interest is to maximize (sometimes even minimize information diffusion in online social media networks. Word-of-mouth (w-o-m communications have been largely employed in a wide range of application scenarios ranging from marketing strategies to creating mass awareness. In recent years, we saw an explosion in the growth of online social networks with many successful service providers like Facebook, twitter,YouTube, LinkedIn etc. ruling the market. The use of online social networks is not limited to fun and building professional networks. Online social networks are now seen as a powerful medium to effect the social and political environment of a country and the world as a whole. Information on these social networking sites sometimes needs to be propagated very effectively and quickly to fulfill the concerned strategies. It is equally important that given some information present on social network, we are able to derive its origin so that rumours can be distinguished from truth. In this paper, we discuss some of the issues and challenges in information diffusion and provenance from online social networks point of view. We also discuss some of the approaches and models that have been used for information diffusion and provenance in online social networks in recent past. Some of these approaches have been evaluated by researchers for specific social network services but we will discuss them in a general sense.

  18. Translocations of amphibians: Proven management method or experimental technique

    Seigel, Richard A.; Dodd, C. Kenneth


    In an otherwise excellent review of metapopulation dynamics in amphibians, Marsh and Trenham (2001) make the following provocative statements (emphasis added): If isolation effects occur primarily in highly disturbed habitats, species translocations may be necessary to promote local and regional population persistence. Because most amphibians lack parental care, they areprime candidates for egg and larval translocations. Indeed, translocations have already proven successful for several species of amphibians. Where populations are severely isolated, translocations into extinct subpopulations may be the best strategy to promote regional population persistence. We take issue with these statements for a number of reasons. First, the authors fail to cite much of the relevant literature on species translocations in general and for amphibians in particular. Second, to those unfamiliar with current research in amphibian conservation biology, these comments might suggest that translocations are a proven management method. This is not the case, at least in most instances where translocations have been evaluated for an appropriate period of time. Finally, the authors fail to point out some of the negative aspects of species translocation as a management method. We realize that Marsh and Trenham's paper was not concerned primarily with translocations. However, because Marsh and Trenham (2001) made specific recommendations for conservation planners and managers (many of whom are not herpetologists or may not be familiar with the pertinent literature on amphibians), we believe that it is essential to point out that not all amphibian biologists are as comfortable with translocations as these authors appear to be. We especially urge caution about advocating potentially unproven techniques without a thorough review of available options.

  19. Multitechnique characterization of lapis lazuli for provenance study.

    Lo Giudice, Alessandro; Re, Alessandro; Calusi, Silvia; Giuntini, Lorenzo; Massi, Mirko; Olivero, Paolo; Pratesi, Giovanni; Albonico, Maria; Conz, Elisa


    Lapis lazuli is one of the oldest precious stone, being used for glyptic as early as 7,000 years ago: jewels, amulets, seals, and inlays are examples of objects produced using this material. Only a few sources of lapis lazuli exist in the world due to the low probability of geological conditions in which it can form, so that the possibility to associate the raw material to man-made objects helps to reconstruct trade routes. Since art objects produced using lapis lazuli are valuable, only nondestructive investigations can be carried out to identify the provenance of the raw materials. Ionoluminescence (IL) is a good candidate for this task. Similar to cathodoluminescence (CL), IL consists in the collection of luminescence spectra induced by megaelectronvolt ion (usually protons) irradiation. The main advantage of IL consists in the possibility of working in air while measuring simultaneously the composition of major and trace elements by means of complementary ion beam analysis techniques like particle-induced X-ray emission (PIXE) or particle-induced gamma-ray emission (PIGE). In the present work, a systematic study of the luminescence properties of lapis lazuli under charged particle irradiation is reported. In the first phase, a multitechnique approach was adopted (CL, scanning electron microscopy with microanalysis, micro-Raman) to characterize luminescent minerals. This characterization was propaedeutic for IL/PIXE/PIGE measurements carried out on significant areas selected on the basis of results obtained previously. Criteria to identify provenance of lapis lazuli from four of the main sources (Afghanistan, Pamir Mountains in Tajikistan, Chile, and Siberia) were proposed.

  20. Evolving provenance in the Proterozoic Pranhita-Godavari Basin, India

    Udeni Amarasinghe; Asru Chaudhuri; Alan S Collins; Gautam Deb; Sarbani Patranabis-Deb


    The Pranhita-Godavari Basin in central eastern India is one of the Proterozoic“Pur?ana”basins of cratonic India. New geochronology demonstrates that it has a vast depositional history of repeated basin reac-tivation from the Palaeoproterozoic to the Mesozoic. U-Pb laser ablation inductively coupled plasma mass spectrometry dating of detrital zircons from two samples of the Somanpalli Groupda member of the oldest sedimentary cycle in the valleydconstrains its depositional age to w1620 Ma and demon-strates a tripartite age provenance with peaks at w3500 Ma, w2480 Ma and w1620 Ma, with minor age peaks in the Eoarchaean (w3.8 Ga) and at w2750 Ma. These ages are consistent with palaeocurrent data suggesting a southerly source from the Krishna Province and Enderby Land in East Antarctica. The similarity in the maximum depositional age with previously published authigenic glauconite ages sug-gest that the origin of the Pranhita-Godvari Graben originated as a rift that formed at a high angle to the coeval evolving late Meosproterozoic Krishna Province as Enderby Land collided with the Dharwar craton of India. In contrast, detrital zircons from the Cycle III Sullavai Group red sandstones yielded a maximum depositional age of 970 ? 20 Ma and had age peaks of w2550 Ma, w1600 Ma and then a number of Mesoproterozoic detrital zircons terminating in three analyses at w970 Ma. The provenance of these is again consistent with a southerly source from the Eastern Ghats Orogen and Antarctica. Later cycles of deposition include the overlying Albaka/Usur Formations and finally the late Palaeozoic to Mesozoic Gondwana Supergroup.

  1. Evolving provenance in the Proterozoic Pranhita-Godavari Basin, India

    Udeni Amarasinghe


    Full Text Available The Pranhita-Godavari Basin in central eastern India is one of the Proterozoic “Purāna” basins of cratonic India. New geochronology demonstrates that it has a vast depositional history of repeated basin reactivation from the Palaeoproterozoic to the Mesozoic. U-Pb laser ablation inductively coupled plasma mass spectrometry dating of detrital zircons from two samples of the Somanpalli Group—a member of the oldest sedimentary cycle in the valley—constrains its depositional age to ∼1620 Ma and demonstrates a tripartite age provenance with peaks at ∼3500 Ma, ∼2480 Ma and ∼1620 Ma, with minor age peaks in the Eoarchaean (∼3.8 Ga and at ∼2750 Ma. These ages are consistent with palaeocurrent data suggesting a southerly source from the Krishna Province and Enderby Land in East Antarctica. The similarity in the maximum depositional age with previously published authigenic glauconite ages suggest that the origin of the Pranhita-Godvari Graben originated as a rift that formed at a high angle to the coeval evolving late Meosproterozoic Krishna Province as Enderby Land collided with the Dharwar craton of India. In contrast, detrital zircons from the Cycle III Sullavai Group red sandstones yielded a maximum depositional age of 970 ± 20 Ma and had age peaks of ∼2550 Ma, ∼1600 Ma and then a number of Mesoproterozoic detrital zircons terminating in three analyses at ∼970 Ma. The provenance of these is again consistent with a southerly source from the Eastern Ghats Orogen and Antarctica. Later cycles of deposition include the overlying Albaka/Usur Formations and finally the late Palaeozoic to Mesozoic Gondwana Supergroup.

  2. c-Src activation promotes nasopharyngeal carcinoma metastasis by inducing the epithelial-mesenchymal transition via PI3K/Akt signaling pathway: a new and promising target for NPC

    Lu, Jinping; Xia, Weixiong; Yu, Yahui; Peng, Yongjian; Wang, Li; Wang, Gang; Ye, Yanfang; Yang, Jing; Liang, Hu; Kang, Tiebang; Lv, Xing


    Aberrant activation of cellular Src (c-Src), a non-receptor tyrosine kinase, could promote cancer progression through activating its downstream signaling pathways. However, the roles of c-Src and phosphorylated-Src (p-Src) in nasopharyngeal carcinoma (NPC) progression are rarely investigated. Herein, we have identified high c-Src concentrations in the serum of NPC patients with distant metastasis using high-throughput protein microarrays. Levels of c-Src in serum and p-Src in human primary NPC samples were unfavorable independent prognostic factors for cancer-specific survival, disease-free survival, and distant metastasis-free survival. Depletion or inactivation of c-Src in NPC cells using sgRNA with CRISPR/Cas9 system or PP2 decreased cell viability, colony formation, migration and invasion in vitro and metastasis in vivo. In contrast, these malignancies could be up-regulated by overexpressed c-Src in a NPC cell line with low-metastasis potential. Furthermore, p-Src was involved in promoting NPC cell metastasis by inducing the epithelial-mesenchymal transition (EMT) process via activating the PI3K/Akt pathway and cytoskeleton remodeling. The p-Src-induced EMT process could be retarded by PP2, which mediated by down-regulating the PI3K/Akt pathway. In conclusion, elevated levels of c-Src in serum and p-Src in primary NPC tissue correlated with poor outcomes of NPC patients. And aberrant activation of c-Src facilitated NPC cells with malignant potential, especially metastasis ability, which mediated by the PI3K/Akt pathway activation and sequentially induced the EMT process. These findings unveiled a promising approach for targeted therapy of advanced NPC. PMID:27078847

  3. PGE2/EP3/SRC signaling induces EGFR nuclear translocation and growth through EGFR ligands release in lung adenocarcinoma cells

    Bazzani, Lorenzo; Donnini, Sandra; Finetti, Federica; Christofori, Gerhard; Ziche, Marina


    Prostaglandin E2 (PGE2) interacts with tyrosine kinases receptor signaling in both tumor and stromal cells supporting tumor progression. Here we demonstrate that in non-small cell lung carcinoma (NSCLC) cells, A549 and GLC82, PGE2 promotes nuclear translocation of epidermal growth factor receptor (nEGFR), affects gene expression and induces cell growth. Indeed, cyclin D1, COX-2, iNOS and c-Myc mRNA levels are upregulated following PGE2 treatment. The nuclear localization sequence (NLS) of EGFR as well as its tyrosine kinase activity are required for the effect of PGE2 on nEGFR and downstream signaling activities. PGE2 binds its bona fide receptor EP3 which by activating SRC family kinases, induces ADAMs activation which, in turn, releases EGFR-ligands from the cell membrane and promotes nEGFR. Amphiregulin (AREG) and Epiregulin (EREG) appear to be involved in nEGFR promoted by the PGE2/EP3-SRC axis. Pharmacological inhibition or silencing of the PGE2/EP3/SRC-ADAMs signaling axis or EGFR ligands i.e. AREG and EREG expression abolishes nEGFR induced by PGE2. In conclusion, PGE2 induces NSCLC cell proliferation by EP3 receptor, SRC-ADAMs activation, EGFR ligands shedding and finally, phosphorylation and nEGFR. Since nuclear EGFR is a hallmark of cancer aggressiveness, our findings reveal a novel mechanism for the contribution of PGE2 to tumor progression. PMID:28415726

  4. High Mobility Group Box Protein 1 Boosts Endothelial Albumin Transcytosis through the RAGE/Src/Caveolin-1 Pathway

    Shang, Dan; Peng, Tao; Gou, Shanmiao; Li, Yiqing; Wu, Heshui; Wang, Chunyou; Yang, Zhiyong


    High-mobility group box protein 1 (HMGB1), an inflammatory mediator, has been reported to destroy cell-cell junctions, resulting in vascular endothelial hyperpermeability. Here, we report that HMGB1 increases the endothelial transcytosis of albumin. In mouse lung vascular endothelial cells (MLVECs), HMGB1 at a concentration of 500 ng/ml or less did not harm cell-cell junctions but rapidly induced endothelial hyperpermeability to 125I-albumin. HMGB1 induced an increase in 125I-albumin and AlexaFluor 488-labeled albumin internalization in endocytosis assays. Depletion of receptor for advanced glycation end products (RAGE), but not TLR2 or TLR4, suppressed HMGB1-induced albumin transcytosis and endocytosis. Genetic and pharmacological destruction of lipid rafts significantly inhibited HMGB1-induced albumin endocytosis and transcytosis. HMGB1 induced the rapid phosphorylation of caveolin (Cav)-1 and Src. Either RAGE gene silencing or soluble RAGE suppressed Cav-1 Tyr14 phosphorylation and Src Tyr418 phosphorylation. The Src inhibitor 4-amino-5-(4-chlorophenyl)-7-(t-butyl) pyrazolo[3,4-d] pyrimidine (PP2) blocked HMGB1-induced Cav-1 Tyr14 phosphorylation. PP2 and overexpression of Cav-1 with a T14F mutation significantly inhibited HMGB1-induced transcytosis and albumin endocytosis. Our findings suggest that HMGB1 induces the transcytosis of albumin via RAGE-dependent Src phosphorylation and Cav-1 phosphorylation. These studies revealed a new mechanism of HMGB1-induced endothelial hyperpermeability. PMID:27572515

  5. Interaction of c-Src with gap junction protein connexin-43. Role in the regulation of cell-cell communication

    Giepmans, B N; Hengeveld, T; Postma, F R; Moolenaar, W H


    Cell-cell communication via connexin-43 (Cx43)-based gap junctions is transiently inhibited by certain mitogens, but the underlying regulatory mechanisms are incompletely understood. Our previous studies have implicated the c-Src tyrosine kinase in mediating transient closure of Cx43-based gap junct

  6. STAT5 activation induced by diabetic LDL depends on LDL glycation and occurs via src kinase activity.

    Brizzi, Maria Felice; Dentelli, Patrizia; Gambino, Roberto; Cabodi, Sara; Cassader, Maurizio; Castelli, Ada; Defilippi, Paola; Pegoraro, Luigi; Pagano, Gianfranco


    Advanced glycation end products (AGEs) have been implicated in the accelerated vascular injury occurring in diabetes. We recently reported that LDL prepared from type 2 diabetic patients (dm-LDL), but not normal LDL (n-LDL) triggered signal transducers and activators of transcription STAT5 activation and p21(waf) expression in endothelial cells (ECs). The aims of the present study were to investigate the role of LDL glycation in dm-LDL- mediated signals and to analyze the molecular mechanisms leading to STAT5 activation. We found that glycated LDL (gly-LDL) triggered STAT5 activation, the formation of a prolactin inducible element (PIE)-binding complex containing STAT5, and increased p21(waf) expression through the activation of the receptor for AGE (RAGE). We also demonstrated that dm-LDL and gly-LDL, but not n-LDL treatment induced the formation of a stable complex containing the activated STAT5 and RAGE. Moreover, gly-LDL triggered src but not JAK2 kinase activity. Pretreatment with the src kinase inhibitor PP1 abrogated both STAT5 activation and the expression of p21(waf) induced by gly-LDL. Consistently, gly-LDL failed to activate STAT5 in src(-/-) fibroblasts. Collectively, our results provide evidence for the role of glycation in dm-LDL-mediated effects and for a specific role of src kinase in STAT5-dependent p21(waf) expression.

  7. SRC burn test in 700-hp oil-designed boiler. Annex Volume A. Southern Research Institute report. Final technical report


    Combustion tests were performed using three forms of Solvent Refined Coal (SRC) as the fuel for a 700 hp oil-designed water-tube boiler at the U.S. Department of Energy (DOE) Pittsburgh Energy Technology Center (PETC). This report contains the results from a program of measurements and analyses performed by Southern Research Institute (SoRI) under contract to the International Coal Refining Company (ICRC). The major objectives of the work performed by Southern Research Institute were: (1) to characterize the particulate matter resulting from the combustion of Solvent Refined Coal (SRC) and its fuel forms, and (2) to develop estimates of the specific collection areas required for varying levels of collection of fly ash from SRC combustion in electrostatic precipitators. The report contains physical and chemical characterizations of particles collected during the combustion experiments, and a discussion of electrostatic precipitation of SRC fly ash based on performance measurements with a small-scale precipitator and on simulations using a mathematical model. 9 references, 90 figures, 14 tables.

  8. Defining the Role of Post-Translational Modifications in SRC-3-Mediated Repression of mRNA Translation


    breast cancer. Balancing immune response: crosstalk between adaptive and innate immune cells during breast cancer progression. Breast Cancer Res, 2007...A, 2009. 106(1): p. 151-6. 15. Wu, R.C., et al., Selective phosphorylations of the SRC-3/AIB1 coactivator integrate genomic reponses to multiple

  9. GD3 synthase overexpression sensitizes hepatocarcinoma cells to hypoxia and reduces tumor growth by suppressing the cSrc/NF-kappaB survival pathway.

    Josep M Lluis

    Full Text Available BACKGROUND: Hypoxia-mediated HIF-1alpha stabilization and NF-kappaB activation play a key role in carcinogenesis by fostering cancer cell survival, angiogenesis and tumor invasion. Gangliosides are integral components of biological membranes with an increasingly recognized role as signaling intermediates. In particular, ganglioside GD3 has been characterized as a proapoptotic lipid effector by promoting cell death signaling and suppression of survival pathways. Thus, our aim was to analyze the role of GD3 in hypoxia susceptibility of hepatocarcinoma cells and in vivo tumor growth. METHODOLOGY/PRINCIPAL FINDINGS: We generated and characterized a human hepatocarcinoma cell line stably expressing GD3 synthase (Hep3B-GD3, which catalyzes the synthesis of GD3 from GM3. Despite increased GD3 levels (2-3 fold, no significant changes in cell morphology or growth were observed in Hep3B-GD3 cells compared to wild type Hep3B cells under normoxia. However, exposure of Hep3B-GD3 cells to hypoxia (2% O(2 enhanced reactive oxygen species (ROS generation, resulting in decreased cell survival, with similar findings observed in Hep3B cells exposed to increasing doses of exogenous GD3. In addition, hypoxia-induced c-Src phosphorylation at tyrosine residues, NF-kappaB activation and subsequent expression of Mn-SOD were observed in Hep3B cells but not in Hep3B-GD3 cells. Moreover, MnTBAP, an antioxidant with predominant SOD mimetic activity, reduced ROS generation, protecting Hep3B-GD3 cells from hypoxia-induced death. Finally, lower tumor growth, higher cell death and reduced Mn-SOD expression were observed in Hep3B-GD3 compared to Hep3B tumor xenografts. CONCLUSION: These findings underscore a role for GD3 in hypoxia susceptibility by disabling the c-Src/NF-kappaB survival pathway resulting in lower Mn-SOD expression, which may be of relevance in hepatocellular carcinoma therapy.

  10. Src, PKCα, and PKCδ are required for αvβ3 integrin-mediated metastatic melanoma invasion

    Bill Heather M


    Full Text Available Abstract Background Integrins, cell-surface receptors that mediate adhesive interactions between cells and the extracellular matrix (ECM, play an important role in cancer progression. Expression of the vitronectin receptor αvβ3 integrin correlates with increased invasive and metastatic capacity of malignant melanomas, yet it remains unclear how expression of this integrin triggers melanoma invasion and metastasis. Results Two melanoma cell lines C8161.9 and M14 both express high levels of αvβ3 integrin and adhere to vitronectin. However, only the highly metastatic C8161.9 cells are capable of invading vitronectin-enriched Matrigel in an αvβ3-depenent manner. Elevated levels of PKCα and PKCδ, and activated Src were detected specifically in the highly metastatic melanoma cells, but not in the low metastatic M14 cells. Inhibition of Src or PKC activity suppressed αvβ3-dependent invasion. Furthermore, over expression of Src or PKCα and PKCδ was sufficient to confer αvβ3-dependent invasiveness to M14 cells. Stress fiber formation and focal adhesion formation were almost completely absent in C8161.9 cells compared to M14 cells. Inhibition of Src signaling was sufficient to restore normal actin architecture, and resulted in decreased p190RhoGAP phosphorylation and enhanced RhoA activity. Src had no effect on Rac activity. Loss of PKCα expression, but not PKCδ, by siRNA inhibited Rac and PAK activity as well as invasiveness. Loss of PKCα restored focal adhesion formation and partially restored stress fiber formation, while loss of PKCδ primarily restored stress fibers. Conclusion The misregulated expression of PKCα and PKCδ and elevated Src activity in metastatic melanoma cells is required for efficient αvβ3-mediated invasion. PKCα and Src enhance αvβ3-mediated invasion in part by increasing the GTPase activity of Rac relative to RhoA. PKCα influences focal adhesion formation, while PKCδ controls stress fibers.

  11. Provenance tracking and querying in the ViroLab Virtual Laboratory

    Balis, B.; Bubak, M.; Pelczar, M.; Wach, J.; Priol, T.; Lefevre, L.; Buyya, R.


    We present an approach to provenance tracking and querying which enables end-users to construct complex queries over provenance records. The use of ontologies for modeling provenance, data and applications enables query construction in an end-user oriented manner, i.e. by using terms of the scientif

  12. Provenance tracking and querying in the ViroLab Virtual Laboratory

    Balis, B.; Bubak, M.; Pelczar, M.; Wach, J.; Priol, T.; Lefevre, L.; Buyya, R.


    We present an approach to provenance tracking and querying which enables end-users to construct complex queries over provenance records. The use of ontologies for modeling provenance, data and applications enables query construction in an end-user oriented manner, i.e. by using terms of the

  13. Provenance tracking and querying in the ViroLab Virtual Laboratory

    Balis, B.; Bubak, M.; Pelczar, M.; Wach, J.; Priol, T.; Lefevre, L.; Buyya, R.


    We present an approach to provenance tracking and querying which enables end-users to construct complex queries over provenance records. The use of ontologies for modeling provenance, data and applications enables query construction in an end-user oriented manner, i.e. by using terms of the scientif

  14. The proto-oncogene c-src is involved in primordial follicle activation through the PI3K, PKC and MAPK signaling pathways.

    Du, Xiao-Yu; Huang, Jian; Xu, Liang-Quan; Tang, Dan-Feng; Wu, Lei; Zhang, Li-Xia; Pan, Xiao-Ling; Chen, Wei-Yun; Zheng, Li-Ping; Zheng, Yue-Hui


    C-src is an evolutionarily conserved proto-oncogene that regulates cell proliferation, differentiation and apoptosis. In our previous studies, we have reported that another proto-oncogene, c-erbB2, plays an important role in primordial follicle activation and development. We also found that c-src was expressed in mammalian ovaries, but its functions in primordial follicle activation remain unclear. The objective of this study is to investigate the role and mechanism of c-src during the growth of primordial follicles. Ovaries from 2-day-old rats were cultured in vitro for 8 days. Three c-src-targeting and one negative control siRNA were designed and used in the present study. PCR, Western blotting and primordial follicle development were assessed for the silencing efficiency of the lentivirus c-src siRNA and its effect on primordial follicle onset. The expression of c-src mRNA and protein in primordial follicle growth were examined using the PCR method and immunohistochemical staining. Furthermore, the MAPK inhibitor PD98059, the PKC inhibitor Calphostin and the PI3K inhibitor LY294002 were used to explore the possible signaling pathways of c-src in primordial folliculogenesis. The results showed that Src protein was distributed in the ooplasmic membrane and the granulosa cell membrane in the primordial follicles, and c-src expression level increased with the growth of primordial follicle. The c-src -targeting lentivirus siRNAs had a silencing effect on c-src mRNA and protein expression. Eight days after transfection of rat ovaries with c-src siRNA, the GFP fluorescence in frozen ovarian sections was clearly discernible under a fluorescence microscope, and its relative expression level was 5-fold higher than that in the control group. Furthermore, the c-src-targeting lentivirus siRNAs lowered its relative expression level 1.96 times. We also found that the development of cultured primordial follicles was completely arrested after c-src siRNA knockdown of c-src

  15. Provenance and sediment fluxes in the Irrawaddy (Ayeyarwadi) River

    Garzanti, Eduardo; Wang, Jiangang; Vezzoli, Giovanni; Limonta, Mara


    The Irrawaddy (Ayeyarwadi) River, still a natural system scarcely affected by human activities, ranks among the five major rivers in the world for its annual suspended load, estimated as 364±60 million tons (Robinson et al., 2007). Sourced in Himalayan glaciers southeast of the eastern Himalayan syntaxis at ca. 28°N, the Irrawaddy originates from the confluence of the Nmai and Mali Rivers, flows southward to receive its major Chindwin tributary in the middle of the central Myanmar Basin, and eventually empties through a nine-armed delta into the Andaman Sea. The compositional fingerprint of bedload sand in the upper Irrawaddy is characterized by common feldspars, medium/high rank of metamorphic rock fragments and high heavy-mineral concentration, reflecting provenance from mid-crustal granitoids, amphibolite-facies and subordinately greenschist-facies rocks widely exposed in the Mogok Belt and Lohit Plutonic Complex. Minor volcanic/metavolcanic and serpentinite grains indicate additional supply from volcanic-arc remnants and the Neotethyan ophiolitic suture. Sand of the Chindwin River has much higher quartz/feldspar ratio and much lower metamorphic indices and heavy-mineral concentration, reflecting provenance mainly from upper crustal sedimentary and very low-grade metasedimentary rocks exposed in the Indo-Burman Ranges (Garzanti et al., 2013). Feldspatho-litho-quartzose to litho-feldspatho-quartzose composition in the lower Irrawaddy is intermediate between that of Chindwin and upper Irrawaddy sand. The slight progressive downstream increase in volcanic rock fragments and chert, and decrease in metamorphic indices, point to additional local supply from volcanic and sedimentary cover rocks. U-Pb age spectra of detrital zircons are characterized by a major cluster between 30 and 150 Ma, corresponding to the long-lasting magmatic activity of the Western Myanmar Arc (Wang et al., 2014), with other clusters at 500-600 Ma and 800-1200 Ma, and a few ages between 1

  16. c-Src modulates estrogen-induced stress and apoptosis in estrogen-deprived breast cancer cells

    Fan, Ping; Griffith, Obi L; Agboke, Fadeke; Anur, Pavana; Zou, Xiaojun; McDaniel, Russell E; Creswell, Karen; Kim, Sung Hoon; Katzenellenbogen, John A; Gray, Joe W; Jordan, V Craig


    The emergence of antiestrogen resistance in breast cancer is an important clinical phenomenon affecting long-term survival in this disease. Identifying factors that convey cell survival in this setting may guide improvements in treatment. Estrogen (E2) can induce apoptosis in breast cancer cells that have been selected for survival after E2 deprivation for long periods (MCF-7:5C cells), but the mechanisms underlying E2-induced stress in this setting have not been elucidated. Here, we report that the c-Src kinase functions as a key adapter protein for the estrogen receptor (ER, ESR1) in its activation of stress responses induced by E2 in MCF-7:5C cells. E2 elevated phosphorylation of c-Src which was blocked by 4-hydroxytamoxifen (4-OHT), suggesting that E2 activated c-Src through the ER. We found that E2 activated the sensors of the unfolded protein response (UPR), IRE1α (ERN1) and PERK kinase (EIF2AK3), the latter of which phosphorylates eukaryotic translation initiation factor-2α (eIF2α). E2 also dramatically increased reactive oxygen species (ROS) production and up-regulated expression of heme oxygenase HO-1 (HMOX1), an indicator of oxidative stress, along with the central energy sensor kinase AMPK (PRKAA2). Pharmacological or RNAi-mediated inhibition of c-Src abolished the phosphorylation of eIF2α and AMPK, blocked E2-induced ROS production, and inhibited E2-induced apoptosis. Together, our results establish that c-Src kinase mediates stresses generated by E2 in long-term E2-deprived cells that trigger apoptosis. This work offers a mechanistic rationale for a new approach in the treatment of endocrine-resistant breast cancer. PMID:23704208

  17. Abnormal cell properties and down-regulated FAK-Src complex signaling in B lymphoblasts of autistic subjects.

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


    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. Copyright © 2011 American Society for Investigative Pathology. Published by Elsevier Inc. All rights reserved.

  18. Nitrotyrosylation of Ca2+ channels prevents c-Src kinase regulation of colonic smooth muscle contractility in experimental colitis.

    Ross, Gracious R; Kang, Minho; Shirwany, Najeeb; Malykhina, Anna P; Drozd, Mary; Akbarali, Hamid I


    Basal levels of c-Src kinase are known to regulate smooth muscle Ca(2+) channels. Colonic inflammation results in attenuated Ca(2+) currents and muscle contraction. Here, we examined the regulation of calcium influx-dependent contractility by c-Src kinase in experimental colitis. Ca(2+)-influx induced contractions were measured by isometric tension recordings of mouse colonic longitudinal muscle strips depolarized by high K(+). The E(max) to CaCl(2) was significantly less in inflamed tissues (38.4 +/- 7.6%) than controls, indicative of reduced Ca(2+) influx. PP2 [4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine], a selective Src kinase inhibitor, significantly reduced the contractile amplitude and shifted the pD(2) from 3.88 to 2.44 in controls, whereas it was ineffective in inflamed tissues (3.66 versus 3.43). After pretreatment with a SIN-1 (3-morpholinosydnonimine)/peroxynitrite combination, the maximal contraction to CaCl(2) was reduced by 46 +/- 7% in controls but unaffected in inflamed tissues (13 +/- 11%). Peroxynitrite also prevented the inhibitory effect of PP2 in control tissues. In colonic single smooth muscle cells, PP2 inhibited Ca(2+) currents by 84.1 +/- 3.9% in normal but only 36.2 +/- 13% in inflamed tissues. Neither the Ca(2+) channel Ca(v)1.2b, gene expression, nor the c-Src kinase activity was altered by inflammation. Western blot analysis showed no change in the Ca(2+) channel protein expression but increased nitrotyrosylated-Ca(2+) channel proteins during inflammation. These data suggest that post-translational modification of Ca(2+) channels during inflammation, possibly nitrotyrosylation, prevents c-Src kinase regulation resulting in decreased Ca(2+) influx.

  19. Src and CXCR4 are involved in the invasiveness of breast cancer cells with acquired resistance to lapatinib.

    De Luca, Antonella; D'Alessio, Amelia; Gallo, Marianna; Maiello, Monica R; Bode, Ann M; Normanno, Nicola


    Lapatinib is a dual EGFR and ErbB-2 tyrosine kinase inhibitor that has significantly improved the clinical outcome of ErbB-2-overexpressing breast cancer patients. However, patients inexorably develop mechanisms of resistance that limit the efficacy of the drug. In order to identify potential targets for therapeutic intervention in lapatinib-resistant patients, we isolated, from ErbB-2-overexpressing SK-Br-3 breast cancer cells, the SK-Br-3 Lap-R-resistant subclone, which is able to routinely grow in 1 µM lapatinib. Resistant cells have a more aggressive phenotype compared with parental cells, as they show a higher ability to invade through a matrigel-coated membrane. Lapatinib-resistant cells have an increased Src kinase activity and persistent levels of activation of ERK1/2 and AKT compared with parental cells. Treatment with the Src inhibitor saracatinib in combination with lapatinib reduces AKT and ERK1/2 phosphorylation and restores the sensitivity of resistant cells to lapatinib. SK-Br-3 Lap-R cells also show levels of expression of CXCR4 that are higher compared with parental cells and are not affected by Src inhibition. Treatment with saracatinib or a specific CXCR4 antibody reduces the invasive ability of SK-Br-3 Lap-R cells, with the two drugs showing cooperative effects. Finally, blockade of Src signaling significantly increases TRAIL-induced cell death in SK-Br-3 Lap-R cells. Taken together, our results demonstrate that breast cancer cells with acquired resistance to lapatinib have a more aggressive phenotype compared with their parental counterpart, and that Src signaling and CXCR4 play an important role in this phenomenon, thus representing potential targets for therapeutic intervention in lapatinib-resistant breast cancer patients.

  20. Src/STAT3-dependent heme oxygenase-1 induction mediates chemoresistance of breast cancer cells to doxorubicin by promoting autophagy.

    Tan, Qixing; Wang, Hongli; Hu, Yongliang; Hu, Meiru; Li, Xiaoguang; Aodengqimuge; Ma, Yuanfang; Wei, Changyuan; Song, Lun


    Chemotherapeutic resistance in breast cancer, whether acquired or intrinsic, remains a major clinical obstacle. Thus, increasing tumor cell sensitivity to chemotherapeutic agents will be helpful in improving the clinical management of breast cancer. In the present study, we found an induction of HO-1 expression in doxorubicin (DOX)-treated MDA-MB-231 human breast adenocarcinoma cells, which showed insensitivity to DOX treatment. Knockdown HO-1 expression dramatically upregulated the incidence of MDA-MB-231 cell death under DOX treatment, indicating that HO-1 functions as a critical contributor to drug resistance in MDA-MB-231 cells. We further observed that DOX exposure induced a cytoprotective autophagic flux in MDA-MB-231 cells, which was dependent on HO-1 induction. Moreover, upregulation of HO-1 expression required the activation of both signal transducer and activator of transcription (STAT)3 and its upstream regulator, protein kinase Src. Abrogating Src/STAT3 pathway activation attenuated HO-1 and autophagy induction, thus increasing the chemosensitivity of MDA-MB-231 cells. Therefore, we conclude that Src/STAT3-dependent HO-1 induction protects MDA-MB-231 breast cancer cells from DOX-induced death through promoting autophagy. In the following study, we further demonstrated the contribution of Src/STAT3/HO-1/autophagy pathway activation to DOX resistance in another breast cancer cell line, MDA-MB-468, which bears a similar phenotype to MDA-MB-231 cells. Therefore, activation of Src/STAT3/HO-1/autophagy signaling pathway might play a general role in protecting certain subtypes of breast cancer cells from DOX-induced cytotoxicity. Targeting this signaling event may provide a potential approach for overcoming DOX resistance in breast cancer therapeutics.

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

    Jennings, Cormac J


    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.

  2. Regulation of voltage-gated sodium current by endogenous Src family kinases in cochlear spiral ganglion neurons in culture.

    Feng, Shuang; Pflueger, Melissa; Lin, Shuang-Xiu; Groveman, Bradley R; Su, Jiping; Yu, Xian-Min


    Voltage-gated sodium (Na+) and potassium (K+)channels have been found to be regulated by Src family kinases(SFKs).However, how these channels are regulated by SFKs in cochlear spiral ganglion neurons (SGNs) remains unknown.Here, we report that altering the activity of endogenous SFKs modulated voltage-gated Na+, but not K+, currents recorded in embryonic SGNs in culture. Voltage-gated Na+ current was suppressed by inhibition of endogenous SFKs or just Src and potentiated by the activation of these enzymes. Detailed investigations showed that under basal conditions, SFK inhibitor application did not significantly affect the voltage-dependent activation, but shifted the steady-state inactivation curves of Na+ currents and delayed the recovery of Na+ currents from inactivation. Application of Src specific inhibitor, Src40–58,not only shifted the inactivation curve but also delayed the recovery of Na+ currents and moved the voltage-dependent activation curve towards the left. The pre-inhibition of SFKs occluded all the effects induced by Src40–58 application, except the left shift of the activation curve. The activation of SFKs did not change either steady-state inactivation or recovery of Na+ currents, but caused the left shift of the activation curve.SFK inhibitor application effectively prevented all the effects induced by SFK activation, suggesting that both the voltage-dependent activation and steady-state inactivation of Na+ current are subjects of SFK regulation. The different effects induced by activation versus inhibition of SFKs implied that under basal conditions, endogenously active and inactive SFKs might be differentially involved in the regulation of voltage-gated Na+ channels in SGNs.

  3. Paleoclimatic, paleovegetational and provenance change in the Ganga Plain during the late Quaternary

    Shailesh Agrawal; Prasanta Sanyal; Melinda K Bera; Jitendra K Dash; Srinivasan Balakrishnan


    Present study aims at reconstructing the paleomonsoonal rainfall, paleovegetation and provenance change during the late Quaternary. Towards this, Bhognipur core, collected from the southern Ganga Plain, have been sampled for soil carbonate (SC) and soil. The 18O values of SC (18OSC) range from −7.6 to −4.9‰. The variations in 18OSC values suggest that during the late Quaternary, the monsoon intensified during MIS 3 and MIS 1 and the maximum lowering of rainfall intensity is observed during MIS 2. The 13C value of SC (13CSC), organic matter dispersed in the soil (13CSOM) and occluded in the carbonate nodules (13CNOM) ranges from −4.1 to +1.4‰, −25.6 to −16.3‰, and −27.7 to −25.0‰, respectively, implies mixed C3–C4 vegetation over the Ganga Plain. Variations in 13CSOM and 13CNOM values at same depth imply preservation problem of pristine organic matter signature. Therefore, it is important to assess the preservation of residual organic matter before using it for paleovegetational reconstruction. The monsoon-vegetation relationship indicates that relative abundances of C3–C4 vegetation were mainly driven by variations in monsoonal rainfall intensity. Using 87Sr/86Sr in SC, we show that the Himalayan river was supplying sediments in the southern part of the Ganga Plain during MIS 3.

  4. Effect of fertilisation on biomass yield, ash and element uptake in SRC willow

    Ugilt Larsen, Søren; Jørgensen, Uffe; Kjeldsen, Jens Bonderup


    Optimal fertilization of short rotation coppice (SRC) willow is important both in terms of economic yield and environmental effect. We measured biomass yield and nutrient uptake in two willow clones, Inger and Tordis, grown on a coarse sandy soil and within six different fertilization regimes....... Fertilization treatments were carried out during two two-year harvest rotations, beginning in the 2nd growth year of the plantation. Willow was fertilized as follows with names referring to type of fertilizer and total quantities of nitrogen (kg ha−1) in first and second year within both rotations: 1) Control0...... related to the quantity of N applied but the effect depended on fertilizer type, harvest rotation and willow clone...

  5. Solvent refined coal (SRC) process. Annual technical progress report, January 1979-December 1979


    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.

  6. Early redox, Src family kinase, and calcium signaling integrate wound responses and tissue regeneration in zebrafish.

    Yoo, Sa Kan; Freisinger, Christina M; LeBert, Danny C; Huttenlocher, Anna


    Tissue injury can lead to scar formation or tissue regeneration. How regenerative animals sense initial tissue injury and transform wound signals into regenerative growth is an unresolved question. Previously, we found that the Src family kinase (SFK) Lyn functions as a redox sensor in leukocytes that detects H(2)O(2) at wounds in zebrafish larvae. In this paper, using zebrafish larval tail fins as a model, we find that wounding rapidly activated SFK and calcium signaling in epithelia. The immediate SFK and calcium signaling in epithelia was important for late epimorphic regeneration of amputated fins. Wound-induced activation of SFKs in epithelia was dependent on injury-generated H(2)O(2). A SFK member, Fynb, was responsible for fin regeneration. This work provides a new link between early wound responses and late regeneration and suggests that redox, SFK, and calcium signaling are immediate "wound signals" that integrate early wound responses and late epimorphic regeneration.

  7. Performance Evaluation of DCA and SRC on a Single Bot Detection

    Al-Hammadi, Yousof; Greensmith, Julie


    Malicious users try to compromise systems using new techniques. One of the recent techniques used by the attacker is to perform complex distributed attacks such as denial of service and to obtain sensitive data such as password information. These compromised machines are said to be infected with malicious software termed a "bot". In this paper, we investigate the correlation of behavioural attributes such as keylogging and packet flooding behaviour to detect the existence of a single bot on a compromised machine by applying (1) Spearman's rank correlation (SRC) algorithm and (2) the Dendritic Cell Algorithm (DCA). We also compare the output results generated from these two methods to the detection of a single bot. The results show that the DCA has a better performance in detecting malicious activities.

  8. Discussion of the therapeutic effects of oncogene c-Src inhibitor on the triple negative breast cancer%癌基因 c-Src 抑制剂对三阴性乳腺癌的疗效评价

    周士波; 姚宇锋


    Objective To explore adjuvant therapy on triple negative breast cancer (TNBC).Methods Develop triple negative breast cancer cell MDA-MB-231 cells,comparing with estrogen receptor (ER)and pro-gesterone receptor (PR)positive T47D cells,and HER2 positive Sk-Br-3 cells.They were treated with the in-hibitor of oncogene c-Src.MTT assay was used to measure cell growth.MTT and immunoelectrophoresis were performed to detect the growth of cells and the change of its signaling pathways.Results The c-Src inhibitor partially inhibited cell growth in T47D.It significantly abolished cell proliferation in triple negative MDA-MB-231 cells.However,it has no inhibitory effect on Sk-Br-3 cells.It was found that therapeutic effects of the c-Src inhibitor were related with the blocking cell growth signaling pathways in different cell lines.The HER2 in-hibitor significantly blocked S phase of cell cycles and cell growth in Sk-Br-3 cells,without any effect on MDA-MB-231 cells.Conclusions Inhibition of the tyrosine kinase activity of c-Src is an effective therapy to treat tri-ple negative breast cancer cells.%目的:探讨癌基因 C-Src 抑制剂对三阴性乳腺癌的辅助治疗作用。方法体外培养三阴性乳腺癌细胞 MDA-MB-231,并与雌激素受体(ER)、孕激素受体(PR)阳性的乳腺癌细胞 T47D 以及HER2阳性的 SK-Br-3细胞做比较。用癌基因 c-Src 抑制剂来治疗三株细胞。采用 MTT 和免疫电泳的方法检测细胞的生长以及细胞信号传导通路的改变。结果c-Src 抑制剂可以部分抑制 T47D 细胞生长,对三阴性乳腺癌细胞 MDA-MB-231有很明显的抑制作用。但是,对 SK-Br-3细胞的生长没有阻滞作用。进一步研究发现,是否抑制细胞生长信号传导通路决定 c-Src 抑制剂的治疗效果。HER2抑制剂可以明显抑制 SK-Br-3细胞周期与生长,而对 MDA-MB-231没有效果。结论抑制癌基因 c-Src 酪氨酸激酶活性对三阴性乳腺癌细胞有很好的治疗效果。

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

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


    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)

  10. Tamoxifen and Src kinase inhibitors as neuroprotective/neuroregenerative drugs after spinal cord injury

    Iris K Salgado; Aranza I Torrado; Jose M Santiago; Jorge D Miranda


    Spinal cord injury (SCI) is a devastating condition that produces signiifcant changes in the life-style of patients. Many molecular and cellular events are triggered after the initial physical impact to the cord. Two major phases have been described in the ifeld of SCI: an acute phase and late phase. Most of the therapeutic strategies are focused on the late phase because this provides an opportunity to target cellular events like apoptosis, demyelination, scar formation and axonal outgrowth. In this mini-review, we will focus on two agents (tamoxifen and a Src kinase family inhibitor known as PP2) that have been shown in our laboratory to produce neuroprotective (increase cell survival) and/or regenerative (axonal outgrowth) actions. The animal model used in our laboratory is adult female rat (~250 g) with a moderate contusion (12.5 mm) to the spinal cord at the T10 level, using the MASCIS impactor device. Tamoxifen or PP2 was administered by implantation of a 15 mg pellet (Innovative Research of America, Sarasota, FL, USA) or by intraperitoneal injections (1.5 mg/kg, every 3 days), respectively, to produce a long-term effect (28 days). Tamoxifen and the Src kinase inhibitor, PP2, are drugs that in rats with a moderate spinal cord injury promote functional locomotor recovery, increase spared white matter tissue, and stimulate axonal outgrowth. Moreover, tamoxifen reduces the formation of reactive oxygen species. Therefore, these drugs are possible therapeutic agents that have a neuroprotective/regen-erative activity in vertebrates with SCI.

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

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


    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.

  12. On the Provenance of Pluto's Nitrogen (N2)

    Singer, Kelsi N


    N2 is abundant in Pluto's atmosphere and on its surface, but the supply is depleted by prodigious atmospheric escape. We demonstrate that cometary impacts could not have delivered enough N2 mass to resupply Pluto's atmospheric escape over time; thus Pluto's N2 is likely endogenous, and therefore was either acquired early in its history or created by chemistry inside/on Pluto. We find that cratering could excavate a considerable amount of N2 to resupply the atmosphere against escape if the near-surface N2 reservoir is deep. However, we find that this process likely falls short of that necessary to resupply the atmosphere against escape by at least an order of magnitude. We conclude that either the escape of N2 from Pluto's atmosphere was on average much lower than the predictions for the current epoch, or that internal activity could be necessary to bring N2 to the surface and resupply escape losses. Observations made by the New Horizons spacecraft in mid-2015 will yield further constraints on the provenance a...

  13. The Dialectal Provenance of London, Wellcome Library, Ms 5262

    Esteban-Segura Laura


    Full Text Available This paper takes into consideration the language found in London, Wellcome Library, MS 5262, a one-volume codex from the early fifteenth century which holds a medical recipe collection. The manuscript, written in Middle English (and with a few fragments in Latin, represents a fine exemplar of a remedybook, a type of writing that has been traditionally considered to be popular. The main aim is to study the dialect of the text contained in folios 3v-61v in order to localise it geographically. The methodology followed for the purpose is grounded on the model supplied by the Linguistic Atlas of Late Mediaeval English (LALME (McIntosh et al. 1986, which consists of several stages including the completion of a survey questionnaire, the creation of the linguistic profile of the text and the application of the ‘fit’-technique (McIntosh et al. 1986, vol. 1: 10-12; Benskin 1991. Extralinguistic features of the manuscript may also be taken into consideration. This comprehensive analysis will help us to circumscribe the dialectal provenance and/or local origin of the text accurately.

  14. Murder by insulin: suspected, purported and proven-a review.

    Marks, Vincent


    Murder by insulin-whether attempted, suspected or proven-is rare. Only 66 cases worldwide could be found for this review. A conviction was secured in 31 cases and additional weapon was employed in 11. Differentiation of attempted homicide from Munchausen syndrome by proxy in the young and from 'mercy killing' in the elderly was not attempted. Most perpetrators were close relatives and most victims were alive when discovered and responded to treatment. Hypoglycaemia is the first clue to homicidal insulin use in living subjects and requires the demonstration of a plasma insulin concentration of generally more than 1000 pmol/L and undetectable plasma C-peptide concentration to establish the diagnosis. Serum glucose measurements are valueless in victims found dead. The presence near the body of insulin vials, syringes or needles, loose talk by the suspected perpetrator or their ready access to insulin may be the only clue. The demonstration of insulin in tissue around an injection site by immunohistopathology or by measuring it in an extract clinches the diagnosis. Immunoassays suitable for clinical use to detect and measure insulin and C-peptide are subject to random errors and cannot be relied upon unless special precautions including separation by gel filtration or HPLC are undertaken prior to analysis. They do not detect or measure accurately a new generation of synthetic insulin analogues. Mass spectrometry will be required to do this and to validate clinical immunoassays, upon which convictions have always had to rely in the past. Copyright 2009 John Wiley & Sons, Ltd.

  15. Provenance of Earth Science Datasets - How Deep Should One Go?

    Ramapriyan, H.; Manipon, G. J. M.; Aulenbach, S.; Duggan, B.; Goldstein, J.; Hua, H.; Tan, D.; Tilmes, C.; Wilson, B. D.; Wolfe, R.; Zednik, S.


    For credibility of scientific research, transparency and reproducibility are essential. This fundamental tenet has been emphasized for centuries, and has been receiving increased attention in recent years. The Office of Management and Budget (2002) addressed reproducibility and other aspects of quality and utility of information from federal agencies. Specific guidelines from NASA (2002) are derived from the above. According to these guidelines, "NASA requires a higher standard of quality for information that is considered influential. Influential scientific, financial, or statistical information is defined as NASA information that, when disseminated, will have or does have clear and substantial impact on important public policies or important private sector decisions." For information to be compliant, "the information must be transparent and reproducible to the greatest possible extent." We present how the principles of transparency and reproducibility have been applied to NASA data supporting the Third National Climate Assessment (NCA3). The depth of trace needed of provenance of data used to derive conclusions in NCA3 depends on how the data were used (e.g., qualitatively or quantitatively). Given that the information is diligently maintained in the agency archives, it is possible to trace from a figure in the publication through the datasets, specific files, algorithm versions, instruments used for data collection, and satellites, as well as the individuals and organizations involved in each step. Such trace back permits transparency and reproducibility.

  16. A global renewable mix with proven technologies and common materials

    Ballabrera, J.; Garcia-Olivares, A.; Garcia-Ladona, E.; Turiel, A.


    A global alternative mix to fossil fuels is proposed, based on proven renewable energy technologies that do not use scarce materials. Taking into account the availability of materials, the resulting mix consists of a combination of onshore and offshore wind turbines, concentrating solar power stations, hydroelectricity and wave power devices attached to the offshore turbines. Solar photovoltaic power could contribute to the mix if its dependence on scarce materials is solved. Material requirements are studied for the generation, power transport and for some future transport systems. The order of magnitude of copper, aluminium, neodymium, lithium, nickel, zinc and platinum that might be required for the proposed solution is obtained and compared with available reserves. While the proposed global alternative to fossil fuels seems technically feasible, lithium, nickel and platinum could become limiting materials for future vehicles fleet if no global recycling system were implemented and rechargeable zinc-air batteries could not be developed. As much as 60% of the current copper reserves would have to be employed in the implementation of the proposed solution. Altogether, the availability of materials may become a long-term physical constraint, preventing the continuation of the usual exponential growth of energy consumption.

  17. Descartes Mountains and Cayley Plains - Composition and provenance

    Drake, M. J.; Taylor, G. J.; Goles, G. G.


    Trace element compositions of petrographically characterized 2-4 mm lithic fragments from Apollo 16 soil samples are used to calculate initial REE concentrations in liquids in equilibrium with lunar anorthosites and to discuss the provenance of the Cayley Formation. Lithic fragments may be subdivided into four groups: (1) ANT rocks, (2) K- and SiO2-rich mesostasis-bearing rocks, (3) poikiloblastic rocks, and (4) (spinel) troctolites. Model liquids in equilibrium with essentially monominerallic anorthosites have initial REE concentrations 5-8 times those of chondrites. The REE contents of K- and SiO2-rich mesostasis-bearing rocks and poikiloblastic rocks are dominated by the mesostasis phases. ANT rocks appear to be more abundant in the Descartes Mountains, while poikiloblastic rocks appear to be more abundant in the Cayley Plains. Poikiloblastic rocks have intermediate to high LIL-element concentrations yet the low gamma-ray activity of Mare Orientale implies low LIL-element concentrations. Consequently, it is unlikely that the Cayley Formation is Orientale ejecta. A local origin as ejecta from smaller impacts is a more plausible model for the deposition of the Cayley Formation.

  18. Experimental evaluation of job provenance in ATLAS environment

    Krenek, A; Sitera, J; Chudoba, J; Dvorak, F; Filipovic, J; KmunIcek, J; Matyska, L; Mulas, M; Ruda, M; Sustr, Z [CESNET, z.s.p.o., Prague (Czech Republic); Campana, S [CERN, Geneva (Switzerland); Molinari, E; Rebatto, D [INFN, Milano (Italy)], E-mail:


    Grid middleware stacks, including gLite, matured into the state of being able to process up to millions of jobs per day. Logging and Bookkeeping, the gLite job-tracking service, keeps pace with this rate; however, it is not designed to provide a long-term archive of information on executed jobs. ATLAS - representative of a large user community - addresses this issue with its own job catalogue (ProdDB). Development of such a customized service, not easily reusable, took considerable effort which is not affordable by smaller communities. On the contrary, Job Provenance (JP), a generic gLite service designed for long-term archiving of information on executed jobs focusing on scalability, extensibility, uniform data view, and configurability, allows more specialized catalogues to be easily built. We present the first results of an experimental JP deployment for the ATLAS production infrastructure where a JP installation was fed with a part of ATLAS jobs, and also stress tested with real production data. The main outcome of this work is a demonstration that JP can complement large-scale application-specific job catalogue services, while serving a similar purpose where there are none available.

  19. Antimicrobial stewardship initiatives throughout Europe: proven value for money

    Edwin J.M. Oberjé


    Full Text Available Antimicrobial stewardship is recognized as a key component to stop the current European spread of antimicrobial resistance. It has also become evident that antimicrobial resistance is a problem that cannot be tackled by single institutions or physicians. Prevention of antimicrobial resistance needs rigorous actions at ward level, institution level, national level and at supra-national levels. Countries can learn from each other and possibly transplant best practices across borders to prevent antimicrobial resistance. The aim of this study is to highlight some of the success stories of proven cost-effective interventions, and to describe the actions that have been taken, the outcomes that have been found, and the difficulties that have been met. In some cases we came across substantial scope for real-life cost savings. Although the best approach to effectively hinder the spread of antimicrobial resistance remains unclear and may vary significantly among settings, several EU-wide examples demonstrate that cost-effective antimicrobial stewardship is possible. Such examples can encourage others to implement (the most cost-effective elements in their system.

  20. Quire: Lightweight Provenance for Smart Phone Operating Systems

    Dietz, Michael; Pisetsky, Yuliy; Shu, Anhei; Wallach, Dan S


    Smartphone apps often run with full privileges to access the network and sensitive local resources, making it difficult for remote systems to have any trust in the provenance of network connections they receive. Even within the phone, different apps with different privileges can communicate with one another, allowing one app to trick another into improperly exercising its privileges (a Confused Deputy attack). In Quire, we engineered two new security mechanisms into Android to address these issues. First, we track the call chain of IPCs, allowing an app the choice of operating with the diminished privileges of its callers or to act explicitly on its own behalf. Second, a lightweight signature scheme allows any app to create a signed statement that can be verified anywhere inside the phone. Both of these mechanisms are reflected in network RPCs, allowing remote systems visibility into the state of the phone when an RPC is made. We demonstrate the usefulness of Quire with two example applications. We built an a...

  1. Opinion: Taking phytoremediation from proven technology to accepted practice.

    Gerhardt, Karen E; Gerwing, Perry D; Greenberg, Bruce M


    Phytoremediation is the use of plants to extract, immobilize, contain and/or degrade contaminants from soil, water or air. It can be an effective strategy for on site and/or in situ removal of various contaminants from soils, including petroleum hydrocarbons (PHC), polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), solvents (e.g., trichloroethylene [TCE]), munitions waste (e.g., 2,4,6-trinitrotoluene [TNT]), metal(loid)s, salt (NaCl) and radioisotopes. Commercial phytoremediation technologies appear to be underutilized globally. The primary objective of this opinion piece is to discuss how to take phytoremediation from a proven technology to an accepted practice. An overview of phytoremediation of soil is provided, with the focus on field applications, to provide a frame of reference for the subsequent discussion on better utilization of phytoremediation. We consider reasons why phytoremediation is underutilized, despite clear evidence that, under many conditions, it can be applied quite successfully in the field. We offer suggestions on how to gain greater acceptance for phytoremediation by industry and government. A new paradigm of phytomanagement, with a specific focus on using phytoremediation as a "gentle remediation option" (GRO) within a broader, long-term management strategy, is also discussed. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  2. Biopsy proven acute interstitial nephritis after treatment with moxifloxacin

    Chatzikyrkou Christos


    Full Text Available Abstract Background Acute interstitial nephritis (AIN is an important cause of reversible acute kidney injury. At least 70% of AIN is caused by various drugs, mainly penicillines and non-steroidal anti-inflammatory drugs. Quinolones are only rarely known to cause AIN and so far cases have been mainly described with older fluoroquinolones. Case Presentation Here we describe a case of biopsy proven interstitial nephritis after moxifloxacin treatment. The patient presented with fever, rigors and dialysis dependent acute kidney injury, just a few days after treatment of a respiratory tract infection with moxifloxacin. The renal biopsy revealed dense infiltrates mainly composed of eosinophils and severe interstitial edema. A course of oral prednisolone (1 mg/kg/day was commenced and rapidly tapered to zero within three weeks. The renal function improved, and the patient was discharged with a creatinine of 107 μmol/l. Conclusion This case illustrates that pharmacovigilance is important to early detect rare side effects, such as AIN, even in drugs with a favourable risk/benefit ratio such as moxifloxacin.

  3. The Skeletal Response to Estrogen is Impaired in Female but not in Male Steroid Receptor Coactivator (SRC)-1 Knock Out Mice

    Mödder, U. I.; Sanyal, A.; Xu, J; O’Malley, B.W.; Spelsberg, T C; Khosla, S.


    Estrogen (E) is critical for the maintenance of bone mass in both female and male mice and steroid receptor coactivator (SRC)-1 has been shown to be important for mediating E effects on bone, at least in female mice. In the present study, we defined the skeletal phenotype of male SRC-1 knock out (KO) mice and compared it with their female littermates. Further, to determine the role of SRC-1 in mediating effects of E on bone in male mice, we examined the skeletal effects of gonadectomy (gnx) w...

  4. Development of SRC-I product analysis. Volume 2. Evaluation of analytical techniques for SRC-I characterization, recycle solvent studies, and product fractionation studies

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


    A data analysis was performed to determine the relationship between the Wilsonville Solvent Quality test result and SRC liquefaction process parameters. The data base studied covers the years 1979 to 1982, Wilsonville runs 133 to 234. Only process-defined material balance data sets were included to best represent steady-state operation. Each material balance period provided 48 variables from which common process conditions were selected by imposing a range of acceptable deviations from a norm, e.g., a reactor hydrogen pressure of 2000 +- 100 psi. Data for all variables vs. solvent quality were plotted, and in some cases variables were compared with each other to determine common trends, e.g. gas production vs. hydrogen consumption. The plotted data produced no discernible trends. Separating the data by coal type (mine location) and identifying common process conditions with coal types still provided no absolute correlations with solvent quality. However, the effect of the weight percent pyrite present in the feed coal produced a consistent trend. A coal containing more than 1.2% pyrite and less than 0.1% sulfate sulfur yielded results in which any one correlation would cluster about a central point. It was observed that, on average, Kentucky Fies and Pyro mine coal and Indiana V coal clustered together, while Kentucky Lafayette and Dotiki mine coals clustered together. These data point clusters for the variables tested were nearly independent of reactor pressure, space rate, and temperature. One unusual observation of all the data points, independent of process conditions, was that at each change of feed coal, the sum of hydrocarbon and heteroatom gas production was greatest for the first 30 days, after which gas production reached a steady state dependent on process conditions, primarily temperature.

  5. Heavy minerals clustering analysis in application of provenance analysis of Kong 2 Member in Kongnan area


    The main task of provenance analysis is to determine the source of sediments and the position of parent rocks. Provenance analysis may find out the relationship between erosion districts and sediment zone, between the uplift and the depression in the process of basin development. The authors use the method of heavy mineral clustering analysis and estimate the provenance direction of Huanghua Depression in the Paleogene Kong 2 Member. Research shows that there were five provenance areas of Kong 2 Member in Kongnan area.They are western (Shenusi), northwestern (Cangzhou), eastern (Ganhuatun), northeastern and southeastern. The main provenance areas were northwestern and western, while the southern provenance could not be ruled out. And these areas are consistent with the known provenance areas.

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

    Ikeda, Robert


    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.

  7. Recovery after prolonged sleep deprivation: residual effects of slow-release caffeine on recovery sleep, sleepiness and cognitive functions.

    Beaumont, Maurice; Batéjat, Denise; Coste, Olivier; Doireau, Philippe; Chauffard, Françoise; Enslen, Marc; Lagarde, Didier; Pierard, Christophe


    A long work schedule often results in sleep deprivation, sleepiness, impaired performance and fatigue. We investigated the residual effects of slow-release caffeine (SRC) on sleep, sleepiness and cognitive performance during a 42-hour recovery period following a 64-hour continuous wakefulness period in 16 healthy males, according to a double-blind, randomised, placebo-controlled, crossover study. Three hundred milligrams of SRC or placebo was given twice a day at 21:00 and 9:00 during the first 48 h of wakefulness. Recovery sleep was analysed with electroencephalography (EEG) and wrist actigraphy, daytime sleepiness with continuous EEG, sleep latency tests and actigraphy and cognitive functions with computerized tests from the NATO AGARD STRES battery. Both drug groups exhibited almost the same sleep architecture with a rebound of slow-wave sleep during both recovery nights and of REM sleep during the second night. Wakefulness level and cognitive functions were similarly impaired in both groups on the first day of recovery and partially returned to baseline on the second. To conclude, SRC appears to have no unwanted side-effects on recovery sleep, wakefulness and cognitive performance after a long period of sleep deprivation and might therefore be a useful choice over other psychostimulants for a long work schedule.

  8. SensePath: Understanding the Sensemaking Process Through Analytic Provenance.

    Nguyen, Phong H; Xu, Kai; Wheat, Ashley; Wong, B L William; Attfield, Simon; Fields, Bob


    Sensemaking is described as the process of comprehension, finding meaning and gaining insight from information, producing new knowledge and informing further action. Understanding the sensemaking process allows building effective visual analytics tools to make sense of large and complex datasets. Currently, it is often a manual and time-consuming undertaking to comprehend this: researchers collect observation data, transcribe screen capture videos and think-aloud recordings, identify recurring patterns, and eventually abstract the sensemaking process into a general model. In this paper, we propose a general approach to facilitate such a qualitative analysis process, and introduce a prototype, SensePath, to demonstrate the application of this approach with a focus on browser-based online sensemaking. The approach is based on a study of a number of qualitative research sessions including observations of users performing sensemaking tasks and post hoc analyses to uncover their sensemaking processes. Based on the study results and a follow-up participatory design session with HCI researchers, we decided to focus on the transcription and coding stages of thematic analysis. SensePath automatically captures user's sensemaking actions, i.e., analytic provenance, and provides multi-linked views to support their further analysis. A number of other requirements elicited from the design session are also implemented in SensePath, such as easy integration with existing qualitative analysis workflow and non-intrusive for participants. The tool was used by an experienced HCI researcher to analyze two sensemaking sessions. The researcher found the tool intuitive and considerably reduced analysis time, allowing better understanding of the sensemaking process.

  9. Infrared reflectance spectra: Effects of particle size, provenance and preparation

    Su, Yin-Fong; Myers, Tanya L.; Brauer, Carolyn S.; Blake, Thomas A.; Forland, Brenda M.; Szecsody, James E.; Johnson, Timothy J.


    We have recently developed methods for making more accurate infrared total and diffuse directional - hemispherical reflectance measurements using an integrating sphere. We have found that reflectance spectra of solids, especially powders, are influenced by a number of factors including the sample preparation method, the particle size and morphology, as well as the sample origin. On a quantitative basis we have investigated some of these parameters and the effects they have on reflectance spectra, particularly in the longwave infrared. In the IR the spectral features may be observed as either maxima or minima: In general, upward-going peaks in the reflectance spectrum result from strong surface scattering, i.e. rays that are reflected from the surface without bulk penetration, whereas downward-going peaks are due to either absorption or volume scattering, i.e. rays that have penetrated or refracted into the sample interior and are not reflected. The light signals reflected from solids usually encompass all such effects, but with strong dependencies on particle size and preparation. This paper measures the reflectance spectra in the 1.3 – 16 micron range for various bulk materials that have a combination of strong and weak absorption bands in order to observe the effects on the spectral features: Bulk materials were ground with a mortar and pestle and sieved to separate the samples into various size fractions between 5 and 500 microns. The median particle size is demonstrated to have large effects on the reflectance spectra. For certain minerals we also observe significant spectral change depending on the geologic origin of the sample. All three such effects (particle size, preparation and provenance) result in substantial change in the reflectance spectra for solid materials; successful identification algorithms will require sufficient flexibility to account for these parameters.

  10. Provenance based data integrity checking and verification in cloud environments.

    Imran, Muhammad; Hlavacs, Helmut; Haq, Inam Ul; Jan, Bilal; Khan, Fakhri Alam; Ahmad, Awais


    Cloud computing is a recent tendency in IT that moves computing and data away from desktop and hand-held devices into large scale processing hubs and data centers respectively. It has been proposed as an effective solution for data outsourcing and on demand computing to control the rising cost of IT setups and management in enterprises. However, with Cloud platforms user's data is moved into remotely located storages such that users lose control over their data. This unique feature of the Cloud is facing many security and privacy challenges which need to be clearly understood and resolved. One of the important concerns that needs to be addressed is to provide the proof of data integrity, i.e., correctness of the user's data stored in the Cloud storage. The data in Clouds is physically not accessible to the users. Therefore, a mechanism is required where users can check if the integrity of their valuable data is maintained or compromised. For this purpose some methods are proposed like mirroring, checksumming and using third party auditors amongst others. However, these methods use extra storage space by maintaining multiple copies of data or the presence of a third party verifier is required. In this paper, we address the problem of proving data integrity in Cloud computing by proposing a scheme through which users are able to check the integrity of their data stored in Clouds. In addition, users can track the violation of data integrity if occurred. For this purpose, we utilize a relatively new concept in the Cloud computing called "Data Provenance". Our scheme is capable to reduce the need of any third party services, additional hardware support and the replication of data items on client side for integrity checking.

  11. Provenance of coastal dune sands along Red Sea, Egypt

    Samir M Zaid


    Texture, mineralogy, and major and trace element geochemistry of 26 coastal dune sand samples were studied to determine the provenance and tectonic environment of two dune fields close to the beaches of Safaga (SF) and Quseir (QS) at the Egyptian Red Sea coast. Onshore winds generate fine, moderate, moderately-well to well-sorted, coarse-skewed to near-symmetrical dune sands with mesokurtic distributions. Winds pick up and transport grains from nearby beach sands and alluvial deposits into a wide Red Sea coastal plain at the border of the beach. The mineralogical (Qt–Ft–Lt) and geochemical composition of the sands, indicate that SF and QS coastal dune sands are mature and influenced by quartz-rich sands. The average CIA values in SF and QS coastal dune sands are low relative to the range of the PAAS, suggesting an arid climate and a low intensity of chemical weathering. The SF and QS coastal dune sand samples are plotted in the recycled orogen and partly in craton interior fields suggesting recycled older sedimentary and partly metamorphic-plutonic sources. The high content of quartz with shell debris and carbonates in coastal dune sands support the recycled sedimentary beach and alluvial sand sources. The dominance of heavy minerals like amphiboles (hornblende) and biotite in the coastal dune sands also supports the effect of metamorphic-plutonic source rocks. The new tectonic discriminant-function diagrams suggest that the coastal dune sands were deposited in a passive margin of a synrift basin. The results provide a good evidence for the extension in the Red Sea rift system during Oligocene-post Pliocene, which is consistent with the general geology of Egypt.

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

    Park Jung-Jin


    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

  13. HIF-1 and c-Src mediate increased glucose uptake induced by endothelin-1 and connexin43 in astrocytes.

    José Carlos Valle-Casuso

    Full Text Available In previous work we showed that endothelin-1 (ET-1 increases the rate of glucose uptake in astrocytes, an important aspect of brain function since glucose taken up by astrocytes is used to supply the neurons with metabolic substrates. In the present work we sought to identify the signalling pathway responsible for this process in primary culture of rat astrocytes. Our results show that ET-1 promoted an increase in the transcription factor hypoxia-inducible factor-1α (HIF-1α in astrocytes, as shown in other cell types. Furthermore, HIF-1α-siRNA experiments revealed that HIF-1α participates in the effects of ET-1 on glucose uptake and on the expression of GLUT-1, GLUT-3, type I and type II hexokinase. We previously reported that these effects of ET-1 are mediated by connexin43 (Cx43, the major gap junction protein in astrocytes. Indeed, our results show that silencing Cx43 increased HIF-1α and reduced the effect of ET-1 on HIF-1α, indicating that the effect of ET-1 on HIF-1α is mediated by Cx43. The activity of oncogenes such as c-Src can up-regulate HIF-1α. Since Cx43 interacts with c-Src, we investigated the participation of c-Src in this pathway. Interestingly, both the treatment with ET-1 and with Cx43-siRNA increased c-Src activity. In addition, when c-Src activity was inhibited neither ET-1 nor silencing Cx43 were able to up-regulate HIF-1α. In conclusion, our results suggest that ET-1 by down-regulating Cx43 activates c-Src, which in turn increases HIF-1α leading to the up-regulation of the machinery required to take up glucose in astrocytes. Cx43 expression can be reduced in response not only to ET-1 but also to various physiological and pathological stimuli. This study contributes to the identification of the signalling pathway evoked after Cx43 down-regulation that results in increased glucose uptake in astrocytes. Interestingly, this is the first evidence linking Cx43 to HIF-1, which is a master regulator of glucose metabolism.

  14. Analysis of the src gene of sarcoma viruses generated by recombination between transformation-defective mutants and quail cellular sequences.

    Wang, L H; Moscovici, C; Karess, R E; Hanafusa, H


    Tumors were produced in quails about 2 months after injection with a transformation-defective mutant of the Schmidt-Ruppin strain of Rous sarcoma virus, subgroup A (SR-A), that retains a small portion of the src gene. Sarcoma viruses were isolated from each of five such tumors. A transformation-defective mutant which has a nearly complete deletion of the src gene was unable to induce tumors. The avian sarcoma viruses recovered from quail tumors (rASV-Q) had biological properties similar to those of the avian sarcoma viruses previously acquired from chicken tumors (rASV-C); these chicken tumors had been induced by the same transformation-defective mutants. Both rASV-Q and rASV-C transformed cells in culture with similar focus morphology and produced tumors within 7 to 14 days after injection into chickens or quails. The size of rASV-Q genomic RNA was indistinguishable from that of SR-A by polyacrylamide gel electrophoresis. The sequences of rASV-Q RNA genomes were analyzed and compared with those of the parental transformation-defective virus, SR-A and of rASV-C by RNase T1 fingerprinting and oligonucleotide mapping. We found that the src sequences of all five isolates of rASV-Q were identical to each other but different from those of SR-A and rASV-C. Of 13 oligonucleotides of rASV-Q identified as src specific, two were not found in either SR-A or rASV-C RNA. Furthermore, some oligonucleotides present in SR-A or rASV-C or both were absent in rASV-Q. No differences were found for the sequences outside the src region in any of the viruses examined. In addition, rASV-Q-infected cells possessed a 60,000-dalton protein specifically precipitable by rabbit serum raised against SR-D-induced tumors. The facts that the src sequences are essentially the same for rASV's recovered from one animal species and different for rASV's obtained from different species provide conclusive evidence that cellular sequences of normal birds were inserted into the viral genome and supplied to

  15. Synthesis, Biological, and Computational Evaluation of Novel 1,3,5-Substituted Indolin-2-one Derivatives as Inhibitors of Src Tyrosine Kinase.

    Kilic-Kurt, Zühal; Bakar, Filiz; Ölgen, Süreyya


    Several substituted indolin-2-one derivatives were synthesized and evaluated for their activities against Src kinase. Several compounds showed activity against Src, with IC50 values in the low micromolar range. Among them, compound 2f showed the most significant activity with an IC50 value of 1.02 μM. Molecular docking studies have been performed for evaluation of the binding modes of compound 2f into the Src active site. The docking structure of compound 2f disclosed that the indole NH forms a hydrogen bond with the carbonyl of Met341. These results suggest that our novel compound 2f is a promising compound for the further development of indole-based drugs targeting Src kinase.

  16. RTK SLAP down: the emerging role of Src-like adaptor protein as a key player in receptor tyrosine kinase signaling.

    Wybenga-Groot, Leanne E; McGlade, C Jane


    SLAP (Src like adaptor protein) contains adjacent Src homology 3 (SH3) and Src homology 2 (SH2) domains closely related in sequence to that of cytoplasmic Src family tyrosine kinases. Expressed most abundantly in the immune system, SLAP function has been predominantly studied in the context of lymphocyte signaling, where it functions in the Cbl dependent downregulation of antigen receptor signaling. However, accumulating evidence suggests that SLAP plays a role in the regulation of a broad range of membrane receptors including members of the receptor tyrosine kinase (RTK) family. In this review we highlight the role of SLAP in the ubiquitin dependent regulation of type III RTKs PDGFR, CSF-1R, KIT and Flt3, as well as Eph family RTKs. SLAP appears to bind activated type III and Eph RTKs via a conserved autophosphorylated juxtamembrane tyrosine motif in an SH2-dependent manner, suggesting that SLAP is important in regulating RTK signaling.

  17. PH006, a novel and selective Src kinase inhibitor, suppresses human breast cancer growth and metastasis in vitro and in vivo.

    Ma, Jin-gui; Huang, He; Chen, Si-meng; Chen, Yi; Xin, Xian-liang; Lin, Li-ping; Ding, Jian; Liu, Hong; Meng, Ling-hua


    The central role of Src in tumor progression and metastasis has validated it as an attractive therapeutic target for the treatment of human breast cancer. The aim of this study was to identify potential Src kinase inhibitor, explore its activity, and mechanism of action in human breast cancer. A strategy integrating focused combinatorial library design, virtual screening, chemical synthesis, and high-throughput screening was adopted and a novel 6-hydrazinopurine-based inhibitor of c-Src kinase PH006 was obtained. The kinase enzymatic activities were measured by enzyme-linked immunosorbent assay. The binding mode between PH006 and Src was profiled by surface plasmon resonance approach and molecular simulation. The anti-proliferative activity was evaluated by Sulforhodamin B (SRB) and Colony formation. The anti-invasion and anti-migration activities were assessed by trans-well and wound healing assay. Results indicated that PH006 was an ATP-competitive Src inhibitor, which selectively inhibited c-Src with an IC₅₀ of 0.38 μM among a panel of 14 diverse tyrosine kinases. PH006 potently inhibited c-Src phosphorylation and c-Src-dependent signal transduction, resulting in inhibition of cell proliferation, migration, and invasion in human breast cancer MDA-MB-231 cells. Further study demonstrated that the anti-proliferative activity of PH006 was ascribed to its capability to arrest cells in G1 phase, while its anti-motility activity was related to suppression of MMP2/9 and HGF secretion. Moreover, PH006 exhibited potent activity against tumor growth as well as metastasis of human breast cancer MDA-MB-435 xenograft beard in nude mice, which was accompanied with reduced Src/FAK signaling in tumor tissue. Taken together, PH006 is a novel selective inhibitor of c-Src and possesses potent activity against breast cancer growth and metastasis, which could be potentially developed as a lead candidate against breast cancers with elevated Src tyrosine kinase activity.

  18. Optimized Combination of Residue Hydrodesulfurization and Residue Fluid Catalytic Cracking

    Chen Junwu


    @@1 Introduction Combination of residue hydrodesulfurization (HDS) and resi-due fluid catalytic cracking (RFCC) is a unique technologyfor processing high-sulfur residue. This paper discusses theoptimized combination of these two processes.

  19. ORCHIDEE-SRC v1.0: an extension of the land surface model ORCHIDEE for simulating short rotation coppice poplar plantations

    De Groote, T.; D. Zona; Broeckx, L. S.; Verlinden, M. S.; Luyssaert, S.; Bellassen, V.; Vuichard, N.; R. Ceulemans; Gobin, A.; Janssens, I. A.


    Modelling biomass production and the environmental impact of short rotation coppice (SRC) plantations is necessary for planning their deployment, as they are becoming increasingly important for global energy production. This paper describes the modification of the widely used land surface model ORCHIDEE for stand-scale simulations of SRC plantations. The model uses weather data, soil texture and species-specific parameters to predict the aboveground (harvestable) biomass...

  20. FDG uptake in the pathologically proven papillary thyroid cancer

    Kim, Tae Sung; Yun, Mi Jin; Cho, Arthur; Lee, Jong Doo [Yonsei University College of Medicine, Seoul (Korea, Republic of)


    Metastatic thyroid cancers with I-131 uptake have been known to show no increase of FDG uptake whereas those without I-131 uptake tend to demonstrate increased uptake on PET. In this study, we evaluated the degree of FDG uptake in primary thyroid cancers of papillary histology before surgery. Forty FDG PET studies were performed on the patients who had papillary cancer proven by fine needle aspiration. The degree of FDG uptake was visually categorized as positive or negative (positive if the tumor showed discernible FDG; negative if the tumor didn't) and the peak standard uptake value (peak SUV) of the papillary thyroid cancer (PTC) were compared with the size of PTC. The mean size of 26 PTC with positive FDG uptake was 1.9{+-} 1.4 cm (0.5 {approx} 5 cm). In 13 PTC with negative FDG uptake, the mean size of those was 0.5 {+-} 0.2 cm (0.2 {approx} 0.9 cm). All PTC larger than 1 cm (2.5 {+-}1.4 cm, 1 {approx} 5 cm) have positive FDG uptake (peak SUV = 6.4 {+-} 5.7, 1.7 {approx} 22.7). Among the micropapillary thyroid cancer (microPTC; PTC smaller than 1 cm), 8 microPTC show positive FDG uptake (peak SUV = 2.9 {+-} 1.3, 1.7 {approx} 5.5), while 13 microPTC show negative finding (peak SUV 1.3 {+-} 0.2, 1.1{approx} 1.7). The size of microPTC with positive FDG uptake is significantly larger than that of microPTC with negative FDG uptake (0.7 {+-} 0.1cm vs 0.4 {+-} 0.2 cm, {rho} = 0.01). All PTCs larger than 1cm show positive FDG uptake in our study. In other words, thyroid lesions larger than 1cm with negative FDG uptake are unlikely to be PTC. So far, only poorly differentiated thyroid cancers are known to show increased FDG uptake. Our results seem to be contradictory to what is known in the literature. Further study is needed to understand better the significance of increased FDG uptake in PTC in relation to expression of NIS and GLUT.

  1. Provenance control on chemical indices of weathering (Taiwan river sands)

    Garzanti, Eduardo; Resentini, Alberto


    Geochemical parameters obtained from the analysis of sediments and sedimentary rocks are widely used to infer weathering and paleo-weathering conditions in source areas. Chemical indices of weathering, however, may not reflect weathering only, or even principally. The concentration of chemical elements in terrigenous sediments is constrained by the original mineralogy of source rocks, and is thus provenance-dependent. Moreover, the mineralogy and consequently the geochemistry of sediments may undergo substantial modifications by diverse physical processes during transport and deposition, including recycling and hydraulic sorting by size, density or shape, and/or by chemical dissolution and precipitation during diagenesis. Around the island of Taiwan, temperature and rainfall are consistently high and relatively homogeneous, and no significant correlation is observed between geochemical and climatic parameters. Physical erosion, fostered by landslides induced by frequent earthquakes and typhoons, prevails because of high relief and extreme rates of tectonic uplift. In such a dynamic orogenic setting, all chemical indices of weathering are controlled principally by the geology of source terranes. Sedimentaclastic and metasedimentaclastic sands carried by western Taiwan rivers draining the pro-wedge display the strongest depletion in Na, Ca, Mg and Sr relative to average upper continental crust, and no depletion or even enrichment in K, Rb and Ba. Low WIP indices reflect erosion of phyllosilicate-dominated rocks in the Slate Belt and extensive recycling of clastic rocks exposed in the Western Foothills. Instead, metamorphiclastic sands carried by eastern Taiwan rivers draining the retro-wedge show no depletion or even enrichment in Mg and Ca, and low CIA and PIA, reflecting contributions from the Tailuko Belt and Coastal Range. Volcaniclastic sands have the same CIA values of their andesitic source rocks (47 ± 1 versus 47 ± 7), indicating that weathering is

  2. 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

    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

  3. TENORM: Coal Combustion Residuals

    Burning coal in boilers to create steam for power generation and industrial applications produces a number of combustion residuals. Naturally radioactive materials that were in the coal mostly end up in fly ash, bottom ash and boiler slag.

  4. Modelling pesticides residues


    This work is a contribution to the development of a specific method to assess the presence of residues in agricultural commodities. The following objectives are formulated: to identify and describe main processes in environment — plant exchanges, to build of a model to assess the residue concentration at harvest in agricultural commodities, to understand the functioning of the modelled system, to characterise pesticides used in field crops and identify optimisation potentials in phytosanitary...

  5. ORCHIDEE-SRC v1.0: an extension of the land surface model ORCHIDEE for simulating short rotation coppice poplar plantations

    T. De Groote


    Full Text Available Modelling biomass production and the environmental impact of short rotation coppice (SRC plantations is necessary for planning their deployment, as they are becoming increasingly important for global energy production. This paper describes the modification of the widely used land surface model ORCHIDEE for stand scale simulations of SRC plantations. The model uses weather data, soil texture and species-specific parameters to predict the aboveground (harvestable biomass production, as well as carbon and energy fluxes of an SRC plantation. Modifications to the model were made to the management, growth, and allocation modules of ORCHIDEE. The modifications presented in this paper were evaluated using data from two poplar based SRC sites. The simulations show that the model performs very well to predict aboveground (harvestable biomass production (within measured ranges, ecosystem photosynthesis (R2 = 0.78, NRMSE = 0.064, PCC = 0.89 and ecosystem respiration (R2 = 0.95, NRMSE = 0.081, PCC = 0.91. Overall, the extended model, ORCHIDEE-SRC, proved to be a tool suitable for predicting biomass production of SRC plantations.

  6. ORCHIDEE-SRC v1.0: an extension of the land surface model ORCHIDEE for simulating short rotation coppice poplar plantations

    De Groote, T.; Zona, D.; Broeckx, L. S.; Verlinden, M. S.; Luyssaert, S.; Bellassen, V.; Vuichard, N.; Ceulemans, R.; Gobin, A.; Janssens, I. A.


    Modelling biomass production and the environmental impact of short rotation coppice (SRC) plantations is necessary for planning their deployment, as they are becoming increasingly important for global energy production. This paper describes the modification of the widely used land surface model ORCHIDEE for stand-scale simulations of SRC plantations. The model uses weather data, soil texture and species-specific parameters to predict the aboveground (harvestable) biomass production, as well as carbon and energy fluxes of an SRC plantation. Modifications to the model were made to the management, growth, and allocation modules of ORCHIDEE. The modifications presented in this paper were evaluated using data from two Belgian poplar-based SRC sites, for which multiple measurements and meteorological data were available. Biomass yield data were collected from 23 other sites across Europe and compared to 22 simulations across a comparable geographic range. The simulations show that the model predicts very well aboveground (harvestable) biomass production (within measured ranges), ecosystem photosynthesis (R2 = 0.78, NRMSE = 0.064, PCC = 0.89) and ecosystem respiration (R2 = 0.95, NRMSE = 0.078 PCC = 0.91). Also soil temperature and soil moisture are simulated adequately, but due to the simplicity of the soil moisture simulation, there are some discrepancies, which also influence the simulation of the latent heat flux. Overall, the extended model, ORCHIDEE-SRC, proved to be a tool suitable for predicting biomass production of SRC plantations.

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

    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


    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.

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

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


    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.

  9. Characterization of promoter region and genomic structure of the murine and human genes encoding Src like adapter protein.

    Kratchmarova, I; Sosinowski, T; Weiss, A; Witter, K; Vincenz, C; Pandey, A


    Src-like adapter protein (SLAP) was identified as a signaling molecule in a yeast two-hybrid system using the cytoplasmic domain of EphA2, a receptor protein tyrosine kinase (Pandey et al., 1995. Characterization of a novel Src-like adapter protein that associates with the Eck receptor tyrosine kinase. J. Biol. Chem. 270, 19201-19204). It is very similar to members of the Src family of cytoplasmic tyrosine kinases in that it contains very homologous SH3 and SH2 domains (Abram and Courtneidge, 2000. Src family tyrosine kinases and growth factor signaling. Exp. Cell. Res. 254, 1-13.). However, instead of a kinase domain at the C-terminus, it contains a unique C-terminal region. In order to exclude the possibility that an alternative form exists, we have isolated genomic clones containing the murine Slap gene as well as the human SLA gene. The coding regions of murine Slap and human SLA genes contain seven exons and six introns. Absence of any kinase domain in the genomic region confirm its designation as an adapter protein. Additionally, we have cloned and sequenced approximately 2.6 kb of the region 5' to the initiator methionine of the murine Slap gene. When subcloned upstream of a luciferase gene, this fragment increased the transcriptional activity about 6-fold in a human Jurkat T cell line and approximately 52-fold in a murine T cell line indicating that this region contains promoter elements that dictate SLAP expression. We have also cloned the promoter region of the human SLA gene. Since SLAP is transcriptionally regulated by retinoic acid and by activation of B cells, the cloning of its promoter region will permit a detailed analysis of the elements required for its transcriptional regulation.

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

    Shinya Takigawa


    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.

  11. Geisinger's ProvenCare methodology: driving performance improvement within a shared governance structure.

    Nolan, Ruth; Wary, Andrea; King, Megan; Laam, Leslie A; Hallick, Susan


    Many performance improvement projects fail because they occur in parallel to the organization's shared governance structure. Leveraging the full potential of its nursing shared governance structure, Geisinger Health System's ProvenCare methodology harnessed the full potential of its staff nurses to create truly reliable workflows that benefit patients and that the team finds professionally satisfying. Using ProvenCare Perinatal and its smoking cessation education intervention and outcomes as an example, the authors describe the ProvenCare methodology.


    Joram M. E. Mbinga


    Full Text Available The phenotypic variation in growth of ten 28-year-old Liquidambar styraciflua provenances was studied at two sites, Lugari and Kakamega in Western Kenya. The trial at each site was established in a Randomized Block Design with the ten provenances replicated in four blocks. Each block had ten square plots corresponding to the ten provenances and each plot had thirty six trees. Analysis of growth data gave results which showed a significant difference (P<0.01 in survival, and growth parameters of tree height, and diameter at breast height (Dbh among provenances at both sites. Ranking based on height growth of the provenances showed the best provenance at Lugari site was Finca Las Victoria-Guatemala, with mean height and Dbh of 35.8 m and 37.1 cm respectively, while the leading provenance at Kakamega site was Tactic Coban, Guatemala, with a mean height and Dbh of 28.7 m and 26.8 cm respectively. The provenance with highest mean survival at Lugari was Finca Las Victoria-Guatemala with a value of 29.2%, while at Kakamega the best provenance was Franklin, Virginia–USA, with a value of 72.2%. Provenance by site interaction was significant as shown by the difference in performance of provenances between the sites. A comparison of the result from the best performing provenances with the growth of the most commonly grown Cupressus lusitanica species in similar sites in Kenya indicates the high potential of L. styraciflua as a commercial plantation species in medium altitude sites in Kenya.

  13. Aberrant trafficking of NSCLC-associated EGFR mutants through the endocytic recycling pathway promotes interaction with Src@

    Band Vimla


    Full Text Available Abstract Background Epidermal growth factor receptor (EGFR controls a wide range of cellular processes, and altered EGFR signaling contributes to human cancer. EGFR kinase domain mutants found in non-small cell lung cancer (NSCLC are constitutively active, a trait critical for cell transformation through activation of downstream pathways. Endocytic trafficking of EGFR is a major regulatory mechanism as ligand-induced lysosomal degradation results in termination of signaling. While numerous studies have examined mutant EGFR signaling, the endocytic traffic of mutant EGFR within the NSCLC milieu remains less clear. Results This study shows that mutant EGFRs in NSCLC cell lines are constitutively endocytosed as shown by their colocalization with the early/recycling endosomal marker transferrin and the late endosomal/lysosomal marker LAMP1. Notably, mutant EGFRs, but not the wild-type EGFR, show a perinuclear accumulation and colocalization with recycling endosomal markers such as Rab11 and EHD1 upon treatment of cells with endocytic recycling inhibitor monensin, suggesting that mutant EGFRs preferentially traffic through the endocytic recycling compartments. Importantly, monensin treatment enhanced the mutant EGFR association and colocalization with Src, indicating that aberrant transit through the endocytic recycling compartment promotes mutant EGFR-Src association. Conclusion The findings presented in this study show that mutant EGFRs undergo aberrant traffic into the endocytic recycling compartment which allows mutant EGFRs to engage in a preferential interaction with Src, a critical partner for EGFR-mediated oncogenesis.

  14. Photometric calibration of NGS/POSS and ESO/SRC plates using the NOAO PDS measuring engine. I - Stellar photometry

    Cutri, Roc M.; Low, Frank J.; Marvel, Kevin B.


    The PDS/Monet measuring engine at the National Optical Astronomy Observatory was used to obtain photometry of nearly 10,000 stars on the NGS/POSS and 2000 stars on the ESO/SRC Survey glass plates. These measurements have been used to show that global transformation functions exist that allow calibration of stellar photometry from any blue or red plate to equivalent Johnson B and Cousins R photoelectric magnitudes. The four transformation functions appropriate for the POSS O and E and ESO/SRC J and R plates were characterized, and it was found that, within the measurement uncertainties, they vary from plate to plate only by photometric zero-point offsets. A method is described to correct for the zero-point shifts and to obtain calibrated B and R photometry of stellar sources to an average accuracy of 0.3-0.4 mag within the range R between values of 8 and 19.5 for red plates in both surveys, B between values of 9 and 20.5 on POSS blue plates, and B between values of 10 and 20.5 on ESO/SRC blue plates. This calibration procedure makes it possible to obtain rapid photometry of very large numbers of stellar sources.

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

    Pene-Dumitrescu Teodora


    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.

  16. Trichothecene mycotoxins activate NLRP3 inflammasome through a P2X7 receptor and Src tyrosine kinase dependent pathway.

    Kankkunen, Päivi; Välimäki, Elina; Rintahaka, Johanna; Palomäki, Jaana; Nyman, Tuula; Alenius, Harri; Wolff, Henrik; Matikainen, Sampsa


    Inflammasome is an intracellular molecular platform of the innate immunity that is a key mediator of inflammation. The inflammasome complex detects pathogens and different danger signals, and triggers cysteine protease caspase-1-dependent processing of pro-inflammatory cytokines IL-1β, and IL-18 in dendritic cells and macrophages. Previously, we have shown that water-damaged building associated trichothecene mycotoxins, including roridin A, trigger IL-1β and IL-18 secretion in human macrophages. However, the molecular basis as well as mechanism behind this trichothecene-induced cytokine secretion has remained uncharacterized. Here, we show that the trichothecene-induced IL-1β secretion is dependent on NLRP3 inflammasome in human primary macrophages. Pharmacological inhibition and small interfering RNA approach showed that the trichothecene-induced NLRP3 inflammasome activation is mediated through ATP-gated P2X7 receptor. Moreover, we show that trichothecene-triggered NLRP3 inflammasome activation is dependent on Src tyrosine kinase activity. In addition, gene silencing of c-Cbl, a negative autophagy-related regulator of c-Src, resulted in enhanced secretion of IL-1β and IL-18 in response to trichothecene mycotoxin stimulation in human macrophages. In conclusion, our results suggest that roridin A, a fungal trichothecene mycotoxin, acts as microbial danger signals that trigger activation of NLRP3 inflammasome through P2X7R and Src tyrosine kinase signaling dependent pathway in human primary macrophages.

  17. Opa+ Neisseria gonorrhoeae exhibits reduced survival in human neutrophils via Src family kinase-mediated bacterial trafficking into mature phagolysosomes.

    Johnson, M Brittany; Ball, Louise M; Daily, Kylene P; Martin, Jennifer N; Columbus, Linda; Criss, Alison K


    During gonorrhoeal infection, there is a heterogeneous population of Neisseria gonorrhoeae (Gc) varied in their expression of opacity-associated (Opa) proteins. While Opa proteins are important for bacterial attachment and invasion of epithelial cells, Opa+ Gc has a survival defect after exposure to neutrophils. Here, we use constitutively Opa- and OpaD+ Gc in strain background FA1090 to show that Opa+ Gc is more sensitive to killing inside adherent, chemokine-treated primary human neutrophils due to increased bacterial residence in mature, degradative phagolysosomes that contain primary and secondary granule antimicrobial contents. Although Opa+ Gc stimulates a potent oxidative burst, neutrophil killing of Opa+ Gc was instead attributable to non-oxidative components, particularly neutrophil proteases and the bactericidal/permeability-increasing protein. Blocking interaction of Opa+ Gc with carcinoembryonic antigen-related cell adhesion molecules (CEACAMs) or inhibiting Src family kinase signalling, which is downstream of CEACAM activation, enhanced the survival of Opa+ Gc in neutrophils. Src family kinase signalling was required for fusion of Gc phagosomes with primary granules to generate mature phagolysosomes. Conversely, ectopic activation of Src family kinases or coinfection with Opa+ Gc resulted in decreased survival of Opa- Gc in neutrophils. From these results, we conclude that Opa protein expression is an important modulator of Gc survival characteristics in neutrophils by influencing phagosome dynamics and thus bacterial exposure to neutrophils' full antimicrobial arsenal.

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

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


    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.

  19. Src kinases and ERK activate distinct responses to Stitcher receptor tyrosine kinase signaling during wound healing in Drosophila.

    Tsarouhas, Vasilios; Yao, Liqun; Samakovlis, Christos


    Metazoans have evolved efficient mechanisms for epidermal repair and survival following injury. Several cellular responses and key signaling molecules that are involved in wound healing have been identified in Drosophila, but the coordination of cytoskeletal rearrangements and the activation of gene expression during barrier repair are poorly understood. The Ret-like receptor tyrosine kinase (RTK) Stitcher (Stit, also known as Cad96Ca) regulates both re-epithelialization and transcriptional activation by Grainy head (Grh) to induce restoration of the extracellular barrier. Here, we describe the immediate downstream effectors of Stit signaling in vivo. Drk (Downstream of receptor kinase) and Src family tyrosine kinases bind to the same docking site in the Stit intracellular domain. Drk is required for the full activation of transcriptional responses but is dispensable for re-epithelialization. By contrast, Src family kinases (SFKs) control both the assembly of a contractile actin ring at the wound periphery and Grh-dependent activation of barrier-repair genes. Our analysis identifies distinct pathways mediating injury responses and reveals an RTK-dependent activation mode for Src kinases and their central functions during epidermal wound healing in vivo.

  20. Leaf morphometric characteristics variability of different beech provenances in juvenile development stage

    Šijačić-Nikolić Mirjana


    Full Text Available The taxonomic status of beech from the Balkan Peninsula is not yet clearly defined. There is no agreement among different authors about the morphological characteristics discriminating between the Balkan and European and/or Eastern beech. For most characteristics, the mean values are different but the ranges of variation overlap considerably. Provenance trial of beech established in Serbia, at the locality Debeli Lug, has provided an opportunity for research of interprovenance variability at the level of leaf morphometric characteristics in juvenile development stage. Research included 10 provenances originating from the Western Balkans (Serbian provenance 36 and 38; Croatian provenance 24 and 25; Bosnian provenance 30 and 32 and from Central Europe (German provenance 47 and 49; Austrian provenance 56 and Hungarian provenance 42, where following morphometric characteristics were analyzed: leaf length (Ll, leaf width (Lw, petiole lenght (Pl, leaf base width on 1 cm (Blw, number of veins - left (Vl, number of veins - right (Vr, distance between 3rd and 4th vein - left (Dv 3-4. The results of this research show existence of clear differentiation among provenances from the Western Balkan and from Central Europe, from the point of leaf dimensions, number of veins and leaf base width. [Projekat Ministarstva nauke Republike Srbije, br. TR31041: Establishment of Wood Plantations Intended for Afforestation of Serbia i br. 43007: Studying climate change and its influence on the environment: impacts, adaptation and mitigation

  1. Growth dynamics variation of different larch provenances under the mountain conditions in Poland

    Kulej, M. [Univ. of Agriculture, Cracow (Poland). Section of Seed Production and Selection


    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

  2. The Application of Cloud Computing to the Creation of Image Mosaics and Management of Their Provenance

    Berriman, G Bruce; Groth, Paul; Juve, Gideon


    We have used the Montage image mosaic engine to investigate the cost and performance of processing images on the Amazon EC2 cloud, and to inform the requirements that higher-level products impose on provenance management technologies. We will present a detailed comparison of the performance of Montage on the cloud and on the Abe high performance cluster at the National Center for Supercomputing Applications (NCSA). Because Montage generates many intermediate products, we have used it to understand the science requirements that higher-level products impose on provenance management technologies. We describe experiments with provenance management technologies such as the "Provenance Aware Service Oriented Architecture" (PASOA).

  3. Proven high-reliability assembly methods applied to avionics fiber-optics high-speed transceivers

    Lauzon, Jocelyn; Leduc, Lorrain; Bessette, Daniel; Bélanger, Nicolas; Larose, Robert; Dion, Bruno


    Harsh environment avionics applications require operating temperature ranges that can extend to, and exceed -50 to 115°C. For obvious maintenance, management and cost arguments, product lifetimes as long as 20 years are also sought. This leads to mandatory long-term hermeticity that cannot be obtained with epoxy or silicone sealing; but only with glass seal or metal solder or brazing. A hermetic design can indirectly result in the required RF shielding of the component. For fiber-optics products, these specifications need to be compatible with the smallest possible size, weight and power consumption. The products also need to offer the best possible high-speed performances added to the known EMI immunity in the transmission lines. Fiber-optics transceivers with data rates per fiber channel up to 10Gbps are now starting to be offered on the market for avionics applications. Some of them are being developed by companies involved in the "normal environment" telecommunications market that are trying to ruggedize their products packaging in order to diversify their customer base. Another approach, for which we will present detailed results, is to go back to the drawing boards and design a new product that is adapted to proven MIL-PRF-38534 high-reliability packaging assembly methods. These methods will lead to the introduction of additional requirements at the components level; such as long-term high-temperature resistance for the fiber-optic cables. We will compare both approaches and demonstrate the latter, associated with the redesign, is the preferable one. The performance of the fiber-optic transceiver we have developed, in terms of qualification tests such as temperature cycling, constant acceleration, hermeticity, residual gaz analysis, operation under random vibration and mechanical shocks and accelerated lifetime tests will be presented. The tests are still under way, but so far, we have observed no performance degradation of such a product after more than

  4. Regulation of the Src kinase-associated phosphoprotein 55 homologue by the protein tyrosine phosphatase PTP-PEST in the control of cell motility.

    Ayoub, Emily; Hall, Anita; Scott, Adam M; Chagnon, Mélanie J; Miquel, Géraldine; Hallé, Maxime; Noda, Masaharu; Bikfalvi, Andreas; Tremblay, Michel L


    PTP-PEST is a cytosolic ubiquitous protein tyrosine phosphatase (PTP) that contains, in addition to its catalytic domain, several protein-protein interaction domains that allow it to interface with several signaling pathways. Among others, PTP-PEST is a key regulator of cellular motility and cytoskeleton dynamics. The complexity of the PTP-PEST interactome underscores the necessity to identify its interacting partners and physiological substrates in order to further understand its role in focal adhesion complex turnover and actin organization. Using a modified yeast substrate trapping two-hybrid system, we identified a cytosolic adaptor protein named Src kinase-associated phosphoprotein 55 homologue (SKAP-Hom) as a novel substrate of PTP-PEST. To confirm PTP-PEST interaction with SKAP-Hom, in vitro pull down assays were performed demonstrating that the PTP catalytic domain and Proline-rich 1 (P1) domain are respectively binding to the SKAP-Hom Y260 and Y297 residues and its SH3 domain. Subsequently, we generated and rescued SKAP-Hom-deficient mouse embryonic fibroblasts (MEFs) with WT SKAP-Hom, SKAP-Hom tyrosine mutants (Y260F, Y260F/Y297F), or SKAP-Hom SH3 domain mutant (W335K). Given the role of PTP-PEST, wound-healing and trans-well migration assays were performed using the generated lines. Indeed, SKAP-Hom-deficient MEFs showed a defect in migration compared with WT-rescued MEFs. Interestingly, the SH3 domain mutant-rescued MEFs showed an enhanced cell migration corresponding potentially with higher tyrosine phosphorylation levels of SKAP-Hom. These findings suggest a novel role of SKAP-Hom and its phosphorylation in the regulation of cellular motility. Moreover, these results open new avenues by which PTP-PEST regulates cellular migration, a hallmark of metastasis.

  5. Silver nanoparticles inhibit VEGF-and IL-1β-induced vascular permeability via Src dependent pathway in porcine retinal endothelial cells

    Park Jongsun


    Full Text Available Abstract The aim of this study is to determine the effects of silver nanoparticles (Ag-NP on vascular endothelial growth factor (VEGF-and interleukin-1 beta (IL-1β-induced vascular permeability, and to detect the underlying signaling mechanisms involved in endothelial cells. Porcine retinal endothelial cells (PRECs were exposed to VEGF, IL-1β and Ag-NP at different combinations and endothelial cell permeability was analyzed by measuring the flux of RITC-dextran across the PRECs monolayer. We found that VEGF and IL-1β increase flux of dextran across a PRECs monolayer, and Ag-NP block solute flux induced by both VEGF and IL-1β. To explore the signalling pathway involved VEGF- and IL-1β-induced endothelial alteration, PRECs were treated with Src inhibitor PP2 prior to VEGF and IL-1β treatment, and the effects were recorded. Further, to clarify the possible involvement of the Src pathways in endothelial cell permeability, plasmid encoding dominant negative(DN and constitutively active(CA form of Src kinases were transfected into PRECs, 24 h prior to VEGF and IL-1β exposure and the effects were recorded. Overexpression of DN Src blocked both VEGF-and IL-1β-induced permeability, while overexpression of CA Src rescues the inhibitory action of Ag-NP in the presence or absence of VEGF and IL-1β. Further, an in vitro kinase assay was performed to identify the presence of the Src phosphorylation at Y419. We report that VEGF and IL-1β-stimulate endothelial permeability via Src dependent pathway by increasing the Src phosphorylation and Ag-NP block the VEGF-and IL-1β-induced Src phosphorylation at Y419. These results demonstrate that Ag-NP may inhibit the VEGF-and IL-1β-induced permeability through inactivation of Src kinase pathway and this pathway may represent a potential therapeutic target to inhibit the ocular diseases such as diabetic retinopathy.

  6. PLC-γ directly binds activated c-Src, which is necessary for carbachol-mediated inhibition of NHE3 activity in Caco-2/BBe cells.

    Zachos, Nicholas C; Lee, Luke J; Kovbasnjuk, Olga; Li, Xuhang; Donowitz, Mark


    Elevated levels of intracellular Ca(2+) ([Ca(2+)]i) inhibit Na(+)/H(+) exchanger 3 (NHE3) activity in the intact intestine. We previously demonstrated that PLC-γ directly binds NHE3, an interaction that is necessary for [Ca(2+)]i inhibition of NHE3 activity, and that PLC-γ Src homology 2 (SH2) domains may scaffold Ca(2+) signaling proteins necessary for regulation of NHE3 activity. [Ca(2+)]i regulation of NHE3 activity is also c-Src dependent; however, the mechanism by which c-Src is involved is undetermined. We hypothesized that the SH2 domains of PLC-γ might link c-Src to NHE3-containing complexes to mediate [Ca(2+)]i inhibition of NHE3 activity. In Caco-2/BBe cells, carbachol (CCh) decreased NHE3 activity by ∼40%, an effect abolished with the c-Src inhibitor PP2. CCh treatment increased the amount of active c-Src as early as 1 min through increased Y(416) phosphorylation. Coimmunoprecipitation demonstrated that c-Src associated with PLC-γ, but not NHE3, under basal conditions, an interaction that increased rapidly after CCh treatment and occurred before the dissociation of PLC-γ and NHE3 that occurred 10 min after CCh treatment. Finally, direct binding to c-Src only occurred through the PLC-γ SH2 domains, an interaction that was prevented by blocking the PLC-γ SH2 domain. This study demonstrated that c-Src 1) activity is necessary for [Ca(2+)]i inhibition of NHE3 activity, 2) activation occurs rapidly (∼1 min) after CCh treatment, 3) directly binds PLC-γ SH2 domains and associates dynamically with PLC-γ under elevated [Ca(2+)]i conditions, and 4) does not directly bind NHE3. Under elevated [Ca(2+)]i conditions, PLC-γ scaffolds c-Src into NHE3-containing multiprotein complexes before dissociation of PLC-γ from NHE3 and subsequent endocytosis of NHE3.

  7. Common themes in PrP signaling: the Src remains the same.

    Ochs, Katharina; Málaga-Trillo, Edward


    The ability of the cellular prion protein (PrP(C)) to trigger intracellular signals appears central to neurodegeneration pathways, yet the physiological significance of such signals is rather puzzling. For instance, PrP(C) deregulation disrupts phenomena as diverse as synaptic transmission in mammals and cell adhesion in zebrafish. Although unrelated, the key proteins in these events -the NMDA receptor (NMDAR) and E-cadherin, respectively- are similarly modulated by the Src family kinase (SFK) Fyn. These observations highlight the importance of PrP(C)-mediated Fyn activation, a finding reported nearly two decades ago. Given their complex functions and regulation, SFKs may hold the key to intriguing aspects of PrP biology such as its seemingly promiscuous functions and the lack of strong phenotypes in knockout mice. Here we provide a mechanistic perspective on how SFKs might contribute to the uncertain molecular basis of neuronal PrP phenotypes affecting ion channel activity, axon myelination and olfactory function. In particular, we discuss SFK target proteins involved in these processes and the role of tyrosine phosphorylation in the regulation of their activity and cell surface expression.

  8. Hierarchical Disabled-1 Tyrosine Phosphorylation in Src family Kinase Activation and Neurite Formation

    Katyal, Sachin; Gao, Zhihua; Monckton, Elizabeth; Glubrecht, Darryl; Godbout, Roseline


    There are two developmentally regulated alternatively spliced forms of Disabled-1 (Dab1) in the chick retina: an early form (Dab1-E) expressed in retinal precursor cells and a late form (Dab1-L) expressed in neuronal cells. The main difference between these two isoforms is the absence of two Src family kinase (SFK) recognition sites in Dab1-E. Both forms retain two Abl/Crk/Nck recognition sites implicated in the recruitment of SH2 domain-containing signaling proteins. One of the Dab1-L-specific SFK recognition sites, at tyrosine(Y)-198, has been shown to be phosphorylated in Reelin-stimulated neurons. Here, we use Reelin-expressing primary retinal cultures to investigate the role of the four Dab1 tyrosine phosphorylation sites on overall tyrosine phosphorylation, Dab1 phosphorylation, SFK activation and neurite formation. We show that Y198 is essential but not sufficient for maximal Dab1 phosphorylation, SFK activation and neurite formation, with Y232 and Y220 playing particularly important roles in SFK activation and neuritogenesis, and Y185 having modifying effects secondary to Y232 and Y220. Our data support a role for all four Dab1 tyrosine phosphorylation sites in mediating the spectrum of activities associated with Reelin-Dab1 signaling in neurons. PMID:17350651

  9. Assessing the potential of short rotation coppice (SRC) for cleanup of radionuclide-contaminated sites.

    Dutton, M V; Humphreys, P N


    A small-scale greenhouse investigation was undertaken using Goat willow (Salix caprea) and aspen (Populus tremula) to evaluate the potential of short rotation coppice for remediation of 137Cs- and 90Sr-contaminated sites. Results showed that both species were able to accumulate these radionuclides from a representative disposal soil (aged) and a spiked soil S. caprea accumulating greater levels of 137Cs than P. tremula, with no difference between species for 90Sr accumulation. For each radionuclide, the distribution in both species was similar, with 137Cs accumulation greatest in the roots, whereas 90Sr accumulation was greatest in the leaves. It was also evident that the soil-to-plant transfer factor (Tf) values for 90Sr were greater than for 137Cs, agreeing with differences in the reported bioavailailablity of these radionuclides in soil Based on the Tf values for S. caprea (conservative), estimated remediation times were 92 and 56 yr, for 137Cs and 90Sr, respectively. It is suggested that the selection of Salix species grown in a system of SRC provides a significant opportunity for removal of both 137Cs and 90Sr, primarily due to its higher biomass production. However, for 137Cs phytoremediation investigations into the appropriate use of soil amendments for increasing bioavailability are required.

  10. VEGF165-induced vascular permeability requires NRP1 for ABL-mediated SRC family kinase activation

    Lampropoulou, Anastasia; Senatore, Valentina; Brash, James T.; Liyanage, Sidath E.; Raimondi, Claudio; Bainbridge, James W.


    The vascular endothelial growth factor (VEGF) isoform VEGF165 stimulates vascular growth and hyperpermeability. Whereas blood vessel growth is essential to sustain organ health, chronic hyperpermeability causes damaging tissue edema. By combining in vivo and tissue culture models, we show here that VEGF165-induced vascular leakage requires both VEGFR2 and NRP1, including the VEGF164-binding site of NRP1 and the NRP1 cytoplasmic domain (NCD), but not the known NCD interactor GIPC1. In the VEGF165-bound receptor complex, the NCD promotes ABL kinase activation, which in turn is required to activate VEGFR2-recruited SRC family kinases (SFKs). These results elucidate the receptor complex and signaling hierarchy of downstream kinases that transduce the permeability response to VEGF165. In a mouse model with choroidal neovascularisation akin to age-related macular degeneration, NCD loss attenuated vessel leakage without affecting neovascularisation. These findings raise the possibility that targeting NRP1 or its NCD interactors may be a useful therapeutic strategy in neovascular disease to reduce VEGF165-induced edema without compromising vessel growth. PMID:28289053

  11. Regulation of Discrete Functional Responses by Syk and Src Family Tyrosine Kinases in Human Neutrophils

    Thornin Ear


    Full Text Available Neutrophils play a critical role in innate immunity and also influence adaptive immune responses. This occurs in good part through their production of inflammatory and immunomodulatory cytokines, in conjunction with their prolonged survival at inflamed foci. While a picture of the signaling machinery underlying these neutrophil responses is now emerging, much remains to be uncovered. In this study, we report that neutrophils constitutively express various Src family isoforms (STKs, as well as Syk, and that inhibition of these protein tyrosine kinases selectively hinders inflammatory cytokine generation by acting posttranscriptionally. Accordingly, STK or Syk inhibition decreases the phosphorylation of signaling intermediates (e.g., eIF-4E, S6K, and MNK1 involved in translational control. By contrast, delayed apoptosis appears to be independent of either STKs or Syk. Our data therefore significantly extend our understanding of which neutrophil responses are governed by STKs and Syk and pinpoint some signaling intermediates that are likely involved. In view of the foremost role of neutrophils in several chronic inflammatory conditions, our findings identify potential molecular targets that could be exploited for future therapeutic intervention.

  12. Role of a hippocampal SRC-family kinase-mediated glutamatergic mechanism in drug context-induced cocaine seeking.

    Xie, Xiaohu; Arguello, Amy A; Wells, Audrey M; Reittinger, Andrew M; Fuchs, Rita A


    Glutamatergic neurotransmission in the dorsal hippocampus (DH) is necessary for drug context-induced reinstatement of cocaine-seeking behavior in an animal model of drug relapse. Furthermore, in vitro studies suggest that the Src family of tyrosine kinases critically regulates glutamatergic cellular functions within the DH. Thus, Src-family kinases in the DH may similarly control contextual cocaine-seeking behavior. To test this hypothesis, rats were trained to lever press for un-signaled cocaine infusions in a distinct context followed by extinction training in a different context. Cocaine-seeking behavior (non-reinforced active lever pressing) was then assessed in the previously cocaine-paired and extinction contexts after AP5 (N-methyl-D-aspartate glutamate (NMDA) receptor (NMDAR) antagonist; 0.25 or 2.5 μg/0.5 μl/hemisphere), PP2 (Src-family kinase inhibitor; 6.25 or 62.5 ng/0.5 μl/hemisphere), Ro25-6981 (NR2B subunit-containing NMDAR antagonist; 0.2 or 2 μg/0.5 μl/hemisphere), or vehicle administration into the DH. Administration of AP5, PP2, or Ro25-6981 into the DH dose-dependently impaired drug context-induced reinstatement of cocaine-seeking behavior relative to vehicle, without altering instrumental behavior in the extinction context or food-reinforced instrumental responding and general motor activity in control experiments. Cocaine-seeking behavior during the first 20 min of the test session in the cocaine-paired context was associated with an increase in NR2B subunit activation, and intra-DH PP2 pretreatment disrupted this relationship. Together, these findings suggest that Src-family kinase activation, NMDAR stimulation, and likely Src-family kinase-mediated NR2B subunit-containing NMDAR activation in the DH are necessary for incentive motivational and/or memory processes that promote contextual cocaine-seeking behavior.

  13. Antiproliferative and pro-apoptotic effects afforded by novel Src-kinase inhibitors in human neuroblastoma cells

    Angelucci Adriano


    Full Text Available Abstract Background Neuroblastoma (NB is the second most common solid malignancy of childhood that usually undergoes rapid progression with a poor prognosis upon metastasis. The Src-family tyrosine kinases (SFKs are a group of proteins involved in cancer development and invasiveness that seem to play an important role in the NB carcinogenesis. Methods To determine cell proliferation, the growth rate was evaluated by both MTT test and cells counted. Analysis of DNA content was performed for the evaluation of the cell cycle and apoptosis. To characterize the mechanisms underlying the antiproliferative effects induced by SI 34, a novel pyrazolo-pyrimidine derivative provided with Src inhibitory activity, the involvement of some cellular pathways that are important for cell proliferation and survival was investigated by western blot assays. In particular, the contribution of cyclins, Src and ERK were examined. Finally, experiments of cell adhesion and invasiveness were performed. Results Treatment of SH-SY5Y human NB cells and CHP100 human neuroepithelioma (NE cultures with three novel pyrazolo[3,4-d]pyrimidine derivatives, namely SI 34, SI 35 and SI 83, inhibits the cell proliferation in a time and concentration-dependent manner. The maximal effect was obtained after 72 hours incubation with SI 34 10 μM. Fluorescence microscopy experiments, flow cytometry analysis and determination of caspase-3 activity by fluorimetric assays showed that SI 34 induced SH-SY5Y apoptosis. Moreover, SI 34 determined cell cycle arrest at the G0/G1 phase, paralleled by a decreased expression of cyclin D1. Furthermore, our data indicate that SI 34 reduces the SH-SY5Y cells adhesion and invasiveness. Evidence that SI 34 inhibits the Src and the ERK-phosphorylation, suggests the mechanism through which it exerts its effects in SH-SY5Y cells. Conclusions Our study shows the ability of this pyrazolo-pyrimidine Src inhibitor in reducing the growth and the invasiveness of

  14. Wood structural differences between northern and southern beech provenances growing at a moderate site.

    Eilmann, B; Sterck, F; Wegner, L; de Vries, S M G; von Arx, G; Mohren, G M J; den Ouden, J; Sass-Klaassen, U


    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

  15. Lowering the Barrier to Reproducible Research by Publishing Provenance from Common Analytical Tools

    Jones, M. B.; Slaughter, P.; Walker, L.; Jones, C. S.; Missier, P.; Ludäscher, B.; Cao, Y.; McPhillips, T.; Schildhauer, M.


    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

  16. Geoscience Australia's enterprise application of provenance standards and systems for physical and digital objects

    Kemp, C.; Car, N. J.


    Geoscience Australia (GA) is a government agency that provides advice on the geology and geography of Australia. It is the custodian of many digital and physical datasets of national significance. For several years GA has been implementing an enterprise approach to provenance management. The goal for transparency and reproducibility for all of GA's information products; an objective supported at the highest levels and explicitly listed in its Science Principles. Currently GA is finalising a set of enterprise tools to assist with provenance management and rolling out provenance reporting to different science areas. GA has adopted or developed: provenance storage systems; provenance collection code libraries (for use within automated systems); reporting interfaces (for manual use) and provenance representation capability within legacy catalogues. Using these tools within GA's science areas involves modelling the scenario first and then assessing whether the area has its data managed in such a way that allows links to data within provenance to be resolvable in perpetuity. We don't just want to represent provenance (demonstrating transparency), we want to access data via provenance (allowing for reproducibility). A subtask of GA's current work is to link physical samples to information products (datasets, reports, papers) by uniquely and persistently identifying samples using International GeoSample Numbers and then modelling automated & manual laboratory workflows and associated tasks, such as data delivery to corporate databases using the W3C's PROV Data Model. We use PROV DM throughout our modelling and systems. We are also moving to deliver all sample and digital dataset metadata across the agency in the Web Ontology Language (OWL) and exposing it via Linked Data methods in order to allow Semantic Web querying of multiple systems allowing provenance to be leveraged using as a single method and query point. Through the Science First Transformation Program GA is

  17. Covariant Residual Entropy

    Hubeny, Veronika E


    A recently explored interesting quantity in AdS/CFT, dubbed 'residual entropy', characterizes the amount of collective ignorance associated with either boundary observers restricted to finite time duration, or bulk observers who lack access to a certain spacetime region. However, the previously-proposed expression for this quantity involving variation of boundary entanglement entropy (subsequently renamed to 'differential entropy') works only in a severely restrictive context. We explain the key limitations, arguing that in general, differential entropy does not correspond to residual entropy. Given that the concept of residual entropy as collective ignorance transcends these limitations, we identify two correspondingly robust, covariantly-defined constructs: a 'strip wedge' associated with boundary observers and a 'rim wedge' associated with bulk observers. These causal sets are well-defined in arbitrary time-dependent asymptotically AdS spacetimes in any number of dimensions. We discuss their relation, spec...

  18. Residual-stress measurements

    Ezeilo, A.N.; Webster, G.A. [Imperial College, London (United Kingdom); Webster, P.J. [Salford Univ. (United Kingdom)


    Because neutrons can penetrate distances of up to 50 mm in most engineering materials, this makes them unique for establishing residual-stress distributions non-destructively. D1A is particularly suited for through-surface measurements as it does not suffer from instrumental surface aberrations commonly found on multidetector instruments, while D20 is best for fast internal-strain scanning. Two examples for residual-stress measurements in a shot-peened material, and in a weld are presented to demonstrate the attractive features of both instruments. (author).

  19. Decomposition of residue currents

    Andersson, Mats; Wulcan, Elizabeth


    Given a submodule $J\\subset \\mathcal O_0^{\\oplus r}$ and a free resolution of $J$ one can define a certain vector valued residue current whose annihilator is $J$. We make a decomposition of the current with respect to Ass$(J)$ that correspond to a primary decomposition of $J$. As a tool we introduce a class of currents that includes usual residue and principal value currents; in particular these currents admit a certain type of restriction to analytic varieties and more generally to construct...

  20. Sharing Residual Liability

    Carbonara, Emanuela; Guerra, Alice; Parisi, Francesco


    Economic models of tort law evaluate the efficiency of liability rules in terms of care and activity levels. A liability regime is optimal when it creates incentives to maximize the value of risky activities net of accident and precaution costs. The allocation of primary and residual liability...... allows policy makers to induce parties to undertake socially desirable care and activity levels. Traditionally, tort law systems have assigned residual liability either entirely on the tortfeasor or entirely on the victim. In this paper, we unpack the cheapest-cost-avoider principle to consider...

  1. A Provenance-Based Infrastructure to Support the Life Cycle of Executable Papers

    Bonnet, Philippe


    of an executable paper. The automated capture of provenance information allows authors to easily integrate and update results into papers as they write, and also helps reviewers better evaluate approaches by enabling them to explore experimental results by varying parameters or data. With a provenance-based system...

  2. Data Provenance Inference in Logic Programming: Reducing Effort of Instance-driven Debugging

    Huq, Mohammad Rezwanul; 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 pro

  3. Plastic Growth response of European beech provenances to dry site conditions

    Stojnic, S.; Sass, U.G.W.; Orlovic, S.; Matovic, B.; Eilmann, B.


    Due to projected global warming, there is a great concern about the ability of European beech to adapt to future climate conditions. Provenance trials provide an excellent basis to assess the potential of various provenances to adjust to given climate conditions. In this study we compared the perfor

  4. Provenance variation in subalpine fir grown as an exotic tree species in Denmark and Iceland

    Skúlason, Brynjar

    Neonectria neomacrospora in Denmark. In Iceland the corkbark fir showed superior results, especially for survival rate and Christmas tree quality. The White River provenance from British Columbia is recommended for use in Denmark. The Mount Taylor provenance from the Cibola National Forest in New Mexico...

  5. nitric oxide triggers the phosphatidylinositol 3-kinase/Akt survival pathway in insulin-producing RINm5F cells by arousing Src to activate insulin receptor substrate-1.

    Tejedo, Juan R; Cahuana, Gladys M; Ramírez, Remedios; Esbert, Margarida; Jiménez, Juan; Sobrino, Francisco; Bedoya, Francisco J


    Mechanisms involved in the protective action of nitric oxide (NO) in insulin-producing cells are a matter of debate. We have previously shown that pharmacological inhibition of c-Src cancels the antiapoptotic action of low and sustained concentrations of exogenous NO. In this study, using insulin-producing RINm5F cells that overexpress Src either permanently active (v-Src) or dominant negative (dn-Src) forms, we determine that this tyrosine kinase is the principal mediator of the protective action of NO. We also show that Src-directed activation of insulin receptor substrate-1, phosphatidylinositol 3-kinase (PI3K), Akt, and Bad phosphorylation conform a substantial component of the survival route because pharmacological inhibition of PI3K and Akt canceled the antiapoptotic effects of NO. Studies performed with the protein kinase G (PKG) inhibitor KT-5823 revealed that NO-dependent activation of c-Src/ insulin receptor substrate-1 is not affected by PKG activation. By contrast, Akt and Bad activation are partially dependent on PKG activation. Endogenous production of NO after overexpression of endothelial nitric oxide synthase in RINm5F cells mimics the effects produced by generation of low amounts of NO from exogenous diethylenetriamine/NO. In addition, we found that NO produces c-Src/PI3K- and PKG-dependent activation of ERK 1/2. The MAPK kinase inhibitor PD 98059 suppresses NO-dependent protection from DNA fragmentation induced by serum deprivation. The protective action of low and sustained concentration of NO is also observed in staurosporine- and Taxol-induced apoptosis. Finally, NO also protects isolated rat islets from DNA fragmentation induced by serum deprivation. These data strengthen the notion that NO production at physiological levels plays a role in protection from apoptosis in pancreatic beta-cells.

  6. Nef alleles from all major HIV-1 clades activate Src-family kinases and enhance HIV-1 replication in an inhibitor-sensitive manner.

    Purushottam S Narute

    Full Text Available The HIV-1 accessory factor Nef is essential for high-titer viral replication and AIDS progression. Nef function requires interaction with many host cell proteins, including specific members of the Src kinase family. Here we explored whether Src-family kinase activation is a conserved property of Nef alleles from a wide range of primary HIV-1 isolates and their sensitivity to selective pharmacological inhibitors. Representative Nef proteins from the major HIV-1 subtypes A1, A2, B, C, F1, F2, G, H, J and K strongly activated Hck and Lyn as well as c-Src to a lesser extent, demonstrating for the first time that Src-family kinase activation is a highly conserved property of primary M-group HIV-1 Nef isolates. Recently, we identified 4-amino substituted diphenylfuropyrimidines (DFPs that selectively inhibit Nef-dependent activation of Src-family kinases as well as HIV replication. To determine whether DFP compounds exhibit broad-spectrum Nef-dependent antiretroviral activity against HIV-1, we first constructed chimeric forms of the HIV-1 strain NL4-3 expressing each of the primary Nef alleles. The infectivity and replication of these Nef chimeras was indistinguishable from that of wild-type virus in two distinct cell lines (U87MG astroglial cells and CEM-T4 lymphoblasts. Importantly, the 4-aminopropanol and 4-aminobutanol derivatives of DFP potently inhibited the replication of all chimeric forms of HIV-1 in both U87MG and CEM-T4 cells in a Nef-dependent manner. The antiretroviral effects of these compounds correlated with inhibition of Nef-dependent activation of endogenous Src-family kinases in the HIV-infected cells. Our results demonstrate that the activation of Hck, Lyn and c-Src by Nef is highly conserved among all major clades of HIV-1 and that selective targeting of this pathway uniformly inhibits HIV-1 replication.

  7. Activation of the FAK-src molecular scaffolds and p130Cas-JNK signaling cascades by alpha1-integrins during colon cancer cell invasion.

    Van Slambrouck, Severine; Grijelmo, Clara; De Wever, Olivier; Bruyneel, Erik; Emami, Shahin; Gespach, Christian; Steelant, Wim F A


    Increased src tyrosine kinase expression and activity has been associated with colon cancer cell invasion and survival. Several signaling pathways are involved in the oncogenic activation of src during the adenoma to carcinoma progression and cellular invasion. In the present study, the synthetic ether lipid analog ET-18-OMe was shown to promote invasion of HCT-8/S11 colon cancer cells into collagen type I through the concomitant activation of src by phosphorylation at Tyr416 (5-30 min) in alpha1-integrin immunoprecipitates containing the integrin binding proteins talin and paxillin, as well as the phoshorylated and activated forms of focal adhesion kinase (FAK) at Tyr397 (a FAK kinase activation signal), Tyr576 and Tyr861. This was associated with the lateral redistribution of alpha1-integrins in focal aggregates and persistent activation of the p130Cas/JNK pathways at 5-30 min, with the subsequent induction and activation of the matrix metalloproteinases MMP-2 and MMP-9 (2-12 h). These activated molecular scaffolds and signaling cascades were not observed in immunoprecipitates of alpha2- and beta1-integrins, and tetraspanin CD9, an invasion and metastasis suppressor linked to integrins and FAK signaling. Our data demonstrate that the lateral redistribution and clustering of alpha1-integrins results in the recruitment of the FAK/src motility-promoting signaling complex involved in cancer cell invasion. Disruption of this proinvasive pathway was accomplished by the dominant negative mutant of src (K295R, kinase dead), src pharmacological inhibitor (PP1) and alpha1-integrin function blocking antibodies. These findings support the notion that the alpha1-integrin- and src-dependent signalosome is a relevant therapeutic target against tumor progression in colon cancer patients.

  8. Diverging Drought Resistance of Scots Pine Provenances Revealed by Infrared Thermography

    Hannes Seidel


    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.

  9. Scientific Reproducibility in Biomedical Research: Provenance Metadata Ontology for Semantic Annotation of Study Description.

    Sahoo, Satya S; Valdez, Joshua; Rueschman, Michael


    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.


    Pipinis, Elias; Stampoulidis, Athanasios; Milios, Elias; Kitikidou, Kyriaki; Radoglou, Kalliopi


    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 (GA3) 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 GA3 (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 GA3 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 GA3 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.

  11. Minimal residual disease diagnostics in acute lymphoblastic leukemia: Need for sensitive, fast, and standardized technologies

    J.J.M. van Dongen (Jacques); V.H.J. van der Velden (Vincent); M. Brüggemann (Monika); A. Orfao (Alberto)


    textabstractMonitoring of minimal residual disease (MRD) has become routine clinical practice in frontline treatment of virtually all childhood acute lymphoblastic leukemia (ALL) and in many adult ALL patients. MRD diagnostics has proven to be the strongest prognostic factor, allowing for risk group

  12. Designing with residual materials

    Walhout, W.; Wever, R.; Blom, E.; Addink-Dölle, L.; Tempelman, E.


    Many entrepreneurial businesses have attempted to create value based on the residual material streams of third parties. Based on ‘waste’ materials they designed products, around which they built their company. Such activities have the potential to yield sustainable products. Many of such companies

  13. Sharing Residual Liability

    Carbonara, Emanuela; Guerra, Alice; Parisi, Francesco


    allows policy makers to induce parties to undertake socially desirable care and activity levels. Traditionally, tort law systems have assigned residual liability either entirely on the tortfeasor or entirely on the victim. In this paper, we unpack the cheapest-cost-avoider principle to consider...

  14. Residual stresses within sprayed coatings

    JIANG Yi; XU Bin-shi; WANG Hai-dou


    Some important developments of residual stress researches for coating-based systems were studied. The following topics were included the sources of residual stresses in coatings: error analysis of Stoney's equation in the curvature method used for the measurement of coating residual stress, the modeling of residual stress and some analytical models for predicting the residual stresses in coatings. These topics should provide some important insights for the fail-safe design of the coating-based systems.

  15. Presenting Provenance Based on User Roles - Experiences from the ACOS System

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


    One goal of provenance is to provide users an understanding of the steps a system took to generate data products. Here, the level of detail captured by provenance becomes an important consideration. As detail is added, more questions can be hypothetically addressed. However, presenting significant provenance detail may also overwhelm end users, for one of two reasons: (i) the detail presented is irrelevant to the objectives, or (ii) the detail requires background knowledge a user lacks. Both of these challenges are present for data generated by the Mauna Loa Solar Observatory’s (MLSO) Advanced Coronal Observing System (ACOS). In ACOS, photometer-based readings are taken of solar activity and subsequently processed into data products consumable by end users. To fully understand these sequences of steps, background knowledge corresponding to various areas (e.g., astronomy, digital imaging, and ACOS specific techniques) is required by end users. This makes reviewing provenance difficult for users outside the ACOS development team, where varying degrees of background may be expected (ranging from outside domain experts in Solar Physics to citizen scientists). Likewise, even when steps taken by ACOS are understandable, they may provide undesired detail to an end user if presented. The work with ACOS involved the development of a Semantic Web based framework to selectively present provenance detail for data products in ACOS. Here, provenance is captured according to two sets of ontologies, the Proof Markup Language, which is an ontology based domain-independent provenance model, and a step ontology, designed to capture hierarchies of provenance steps. Used in combination, these ontology sets enable the creation of multiple levels of provenance, ranging from coarse to fine grained detail. In this setting, users may choose to expand/collapse provenance steps to view desired details. However, the specific provenance details a user initially sees is defined through

  16. Steam/fuel system optimization report: 6000-tpd SRC-I Demonstration Plant

    Vakil, T.D.


    The design and configuration of the steam and fuel system for the 6000-ton-per-day (tpd) SRC-I Demonstration plant have been optimized, based on requirements for each area of the plant that were detailed in Area Baseline Designs of December 1982. The system was optimized primarily for the two most likely modes of plant operation, that is, when the expanded-bed hydrocracker (EBH) is operating at either high or low conversion, with all other units operating. However, the design, as such, is also operable under four other anticipated operating modes. The plant is self-sufficient in fuel except when the coker/calciner unit is not operating; then the required fuel oil import ranges from 80 to 125 MM Btu/h, lower heating value (LHV). The system affords stable operation under varying fuel gas availability and is reliable, flexible, and efficient. The optimization was based on maximizing overall efficiency of the steam system. The system was optimized to operate at five different steam-pressure levels, which are justifiable based on the plant's team requirements for process, heat duty, and power. All identified critical equipment drives will be run by steam turbines. Also part of the optimization was elimination of the steam evaporator in the wastewater treatment area. This minimized the impact on the steam system of operating in either the discharge of zero-discharge mode; the steam system remains essentially the same for either mode. Any further optimization efforts should be based on overall cost-effectiveness.

  17. Effects of inhalation exposure to SRC-II heavy and middle distillates

    Springer, D.L.; Miller, R.A.; Weimer, W.C.; Ragan, H.A.; Buschbom, R.L.; Mahlum, D.D.


    To expand the data base on potential health effects of coal liquefaction materials, we have performed studies with both solvent refined coal (SRC)-II heavy distillate (HD) and middle distillate (MD). Weight gain for exposed animals was less than that of controls and was dose-related, ranging from no significant difference for animals in the low-exposure group to failure to gain in the high-dose animals. Liver weights increased significantly over controls, and thymus weights decreased for animals sacrificed at 5 and 13 weeks. After both exposure periods, there were significant treatment-related decreases in erythrocyte parameters and in certain types of white blood cells (WBC). Bone marrow cellularity, and numbers of megakaryocytes consistently decreased, suggesting that bone marrow is a target tissue for high-boiling coal liquids. Microscopic evaluation of tissue indicated exposure-related changes is listed. In contrast to the reported mutagenic and carcinogenic effects observed for the high-boiling coal liquids, middle-boiling-range materials lacked such activity in these assays. These data demonstrate a great deal of similarity in the kinds of effects observed following exposure to middle- and high-boiling-range coal liquids. However, the significance of changes in organ weights and peripheral blood parameters are not always readily apparent following a subchronic study. Because of this, we exposed animals to HD in a manner similar to that for the subchronic experiment and have followed these animals throughout their lives for the development of adverse effects such as reduced longevity and the appearance of tumors. Results from this study will be available for mice in FY 1985 and for rats in FY 1986.

  18. Odin (ANKS1A is a Src family kinase target in colorectal cancer cells

    Feller Stephan M


    Full Text Available Abstract Background Src family kinases (SFK are implicated in the development of some colorectal cancers (CRC. One SFK member, Lck, is not detectable in normal colonic epithelium, but becomes aberrantly expressed in a subset of CRCs. Although SFK have been extensively studied in fibroblasts and different types of immune cells, their physical and functional targets in many epithelial cancers remain poorly characterised. Results 64 CRC cell lines were tested for expression of Lck. SW620 CRC cells, which express high levels of Lck and also contain high basal levels of tyrosine phosphorylated (pY proteins, were then analysed to identify novel SFK targets. Since SH2 domains of SFK are known to often bind substrates after phosphorylation by the kinase domain, the LckSH2 was compared with 14 other SH2s for suitability as affinity chromatography reagent. Mass spectrometric analyses of LckSH2-purified pY proteins subsequently identified several proteins readily known as SFK kinase substrates, including cortactin, Tom1L1 (SRCASM, GIT1, vimentin and AFAP1L2 (XB130. Additional proteins previously reported as substrates of other tyrosine kinase were also detected, including the EGF and PDGF receptor target Odin. Odin was further analysed and found to contain substantially less pY upon inhibition of SFK activity in SW620 cells, indicating that it is a formerly unknown SFK target in CRC cells. Conclusion Rapid identification of known and novel SFK targets in CRC cells is feasible with SH2 domain affinity chromatography. The elucidation of new SFK targets like Odin in epithelial cancer cells is expected to lead to novel insight into cancer cell signalling mechanisms and may also serve to indicate new biomarkers for monitoring tumor cell responses to drug treatments.