Distinct phosphorylation requirements regulate cortactin activation by TirEPEC and its binding to N-WASP

<p>Abstract</p> <p>Background</p> <p>Cortactin activates the actin-related 2/3 (Arp2/3) complex promoting actin polymerization to remodel cell architecture in multiple processes (e.g. cell migration, membrane trafficking, invadopodia formation etc.). Moreover, it was called the Achilles' heel of the actin cytoskeleton because many pathogens hijack signals that converge on this oncogenic scaffolding protein. Cortactin is able to modulate N-WASP activation <it>in vitro </it>in a phosphorylation-dependent fashion. Thus Erk-phosphorylated cortactin is efficient in activating N-WASP through its SH3 domain, while Src-phosphorylated cortactin is not. This could represent a switch on/off mechanism controlling the coordinated action of both nucleator promoting factors (NPFs). Pedestal formation by enteropathogenic <it>Escherichia coli </it>(EPEC) requires N-WASP activation. N-WASP is recruited by the cell adapter Nck which binds a major tyrosine-phosphorylated site of a bacterial injected effector, Tir (translocated intimin receptor). Tir-Nck-N-WASP axis defines the current major pathway to actin polymerization on pedestals. In addition, it was recently reported that EPEC induces tyrosine phosphorylation of cortactin.</p> <p>Results</p> <p>Here we demonstrate that cortactin phosphorylation is absent on N-WASP deficient cells, but is recovered by re-expression of N-WASP. We used purified recombinant cortactin and Tir proteins to demonstrate a direct interaction of both that promoted Arp2/3 complex-mediated actin polymerization <it>in vitro</it>, independently of cortactin phosphorylation.</p> <p>Conclusion</p> <p>We propose that cortactin binds Tir through its N-terminal part in a tyrosine and serine phosphorylation independent manner while SH3 domain binding and activation of N-WASP is regulated by tyrosine and serine mediated phosphorylation of cortactin. Therefore cortactin could act on Tir-Nck-N-WASP pathway and control a possible cycling activity of N-WASP underlying pedestal formation.</p

In Situ Compatibilization of Biopolymer Ternary Blends by Reactive Extrusion with Low-Functionality Epoxy-Based Styrene Acrylic Oligomer

[EN] The present study reports on the use of low-functionality epoxy-based styrene¿acrylic oligomer (ESAO) to compatibilize immiscible ternary blends made of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), polylactide (PLA), and poly(butylene adipate-co-terephthalate) (PBAT). The addition during melt processing of low-functionality ESAO at two parts per hundred resin (phr) of biopolymer successfully changed the soften inclusion phase in the blend system to a thinner morphology, yielding biopolymer ternary blends with higher mechanical ductility and also improved oxygen barrier performance. The compatibilization achieved was ascribed to the in situ formation of a newly block terpolymer, i.e. PHBVb- PLA-b-PBAT, which was produced at the blend interface by the reaction of the multiple epoxy groups present in ESAO with the functional terminal groups of the biopolymers. This chemical reaction was mainly linear due to the inherently low functionality of ESAO and the more favorable reactivity of the epoxy groups with the carboxyl groups of the biopolymers, which avoided the formation of highly branched and/or cross-linked structures and thus facilitated the films processability. Therefore, the reactive blending of biopolymers at different mixing ratios with low-functionality ESAO represents a straightforward methodology to prepare sustainable plastics at industrial scale with different physical properties that can be of interest in, for instance, food packaging applications.This research was funded by the EU H2020 project YPACK (Reference number 773872) and by the Spanish Ministry of Science, Innovation, and Universities (MICIU) with project numbers MAT2017-84909-C2-2-R and AGL2015-63855-C2-1-R. L. Quiles-Carrillo wants to thank the Spanish Ministry of Education, Culture, and Sports (MECD) for financial support through his FPU Grant Number FPU15/03812. Torres-Giner also acknowledges the MICIU for his Juan de la Cierva contract (IJCI-2016-29675).Quiles-Carrillo, L.; Montanes, N.; Lagaron, J.; Balart, R.; Torres-Giner, S. (2019). In Situ Compatibilization of Biopolymer Ternary Blends by Reactive Extrusion with Low-Functionality Epoxy-Based Styrene Acrylic Oligomer. Journal of Polymers and the Environment. 27(1):84-96. https://doi.org/10.1007/s10924-018-1324-2S8496271Babu RP, O’Connor K, Seeram R (2013) Prog Biomater 2:8Torres-Giner S, Torres A, Ferrándiz M, Fombuena V, Balart R (2017) J Food Saf 37:e12348Quiles-Carrillo L, Montanes N, Boronat T, Balart R, Torres-Giner S (2017) Polym Test 61:421Zakharova E, Alla A, Martínez A, De Ilarduya S, Muñoz-Guerra (2015) RSC Adv 5:46395Steinbüchel A, Valentin HE (1995) FEMS Microbiol Lett 128:219McChalicher CWJ, Srienc F (2007) J Biotechnol 132:296Reis KC, Pereira J, Smith AC, Carvalho CWP, Wellner N, Yakimets I (2008) J Food Eng 89:361Vink ETH, Davies S (2015) Ind Biotechnol 11:167John RP, Nampoothiri KM, Pandey A (2006) Process Biochem 41:759Madhavan Nampoothiri K, Nair NR, John RP (2010) Biores Technol 101:8493Garlotta D (2001) J Polym Environ 9:63Lim LT, Auras R, Rubino M (2008) Prog Polym Sci 33:820Quiles-Carrillo L, Montanes N, Sammon C, Balart R, Torres-Giner S (2018) Ind Crops Prod 111:878Quiles-Carrillo L, Blanes-Martínez MM, Montanes N, Fenollar O, Torres-Giner S, Balart R (2018) Eur Polym J 98:402Witt U, Müller R-J, Deckwer W-D (1997) J Environ Polym Degrad 5:81Siegenthaler KO, Künkel A, Skupin G, Yamamoto M (2012) Ecoflex® and Ecovio®: biodegradable, performance-enabling plastics. In: Rieger B, Künkel A, Coates GW, Reichardt R, Dinjus E, Zevaco TA (eds) Synthetic biodegradable polymers. Springer, Berlin Heidelberg, p 91Jiang L, Wolcott MP, Zhang J (2006) Biomacromol 7:199Brandelero RPH, Yamashita F, Grossmann MVE (2010) Carbohyd Polym 82:1102Muthuraj R, Misra M, Mohanty AK (2014) J Polym Environ 22:336Porter RS, Wang L-H (1992) Polymer 33(10): 2019Koning C, Van Duin M, Pagnoulle C, Jerome R (1998) Prog Polym Sci 23:707Muthuraj R, Misra M, Mohanty AK (2017) J Appl Polym Sci 135:45726Ryan AJ (2002) Nat Mater 1:8Wu D, Zhang Y, Yuan L, Zhang M, Zhou W (2010) J Polym Sci Part B 48:756Kim CH, Cho KY, Choi EJ, Park JK (2000) J Appl Polym Sci 77:226Supthanyakul R, Kaabbuathong N, Chirachanchai S (2016) Polymer 105:1Na Y-H, He Y, Shuai X, Kikkawa Y, Doi Y, Inoue Y (2002) Biomacromolecules 3:1179Zeng J-B, Li K-A, Du A-K (2015) RSC Adv 5:32546Xanthos M, Dagli SS (1991) Polym Eng Sci 31:929Sundararaj U, Macosko CW (1995) Macromolecules 28:2647Milner ST, Xi H (1996) J Rheol 40:663Villalobos M, Awojulu A, Greeley T, Turco G, Deeter G (2006) Energy 31:3227Torres-Giner S, Montanes N, Boronat T, Quiles-Carrillo L, Balart R (2016) Eur Polym J 84:693Lehermeier HJ, Dorgan JR (2001) Polym Eng Sci 41:2172Liu B, Xu Q (2013) J Mater Sci Chem Eng 1:9Eslami H, Kamal MR (2013) J Appl Polym Sci 129:2418Loontjens T, Pauwels K, Derks F, Neilen M, Sham CK, Serné M (1997) J Appl Polym Sci 65:1813Ojijo V, Ray SS (2015) Polymer 80:1Frenz V, Scherzer D, Villalobos M, Awojulu AA, Edison M, Van Der Meer R (2008) Multifunctional polymers as chain extenders and compatibilizers for polycondensates and biopolymers. In: Technical papers, regional technical conference—society of plastics engineers, p. 3/1678Utracki LA (2002) Can J Chem Eng 80:1008Al-Itry R, Lamnawar K, Maazouz A (2012) Polym Degrad Stab 97:1898Lin S, Guo W, Chen C, Ma J, Wang B (2012) Mater Des (1980–2015) 36: 604Arruda LC, Magaton M, Bretas RES, Ueki MM (2015) Polym Test 43:27Wang Y, Fu C, Luo Y, Ruan C, Zhang Y, Fu Y (2010) J Wuhan Univ Technol Mater Sci Ed 25:774Wei D, Wang H, Xiao H, Zheng A, Yang Y (2015) Carbohyd Polym 123:275Abdelwahab MA, Taylor S, Misra M, Mohanty AK (2015) Macromol Mater Eng 300:299Sun Q, Mekonnen T, Misra M, Mohanty AK (2016) J Polym Environ 24:23Torres-Giner S, Gimeno-Alcañiz JV, Ocio MJ, Lagaron JM (2011) J Appl Polym Sci 122:914Miyata T, Masuko T (1998) Polymer 39:5515Muthuraj R, Misra M, Mohanty AK (2015) J Appl Polym Sci 132:42189Ren J, Fu H, Ren T, Yuan W (2009) Carbohyd Polym 77:576Torres-Giner S, Montanes N, Fenollar O, García-Sanoguera D, Balart R (2016) Mater Des 108:648Jamshidian M, Tehrany EA, Imran M, Jacquot M, Desobry S (2010) Compr Rev Food Sci Food Saf 9:552Savenkova L, Gercberga Z, Nikolaeva V, Dzene A, Bibers I, Kalnin M (2000) Process Biochem 35:573Costa ARM, Almeida TG, Silva SML, Carvalho LH, Canedo EL (2015) Polym Test 42:115Zhang K, Mohanty AK, Misra M (2012) ACS Appl Mater Interfaces 4:3091Zhang N, Wang Q, Ren J, Wang L (2009) J Mater Sci 44:250Chinsirikul W, Rojsatean J, Hararak B, Kerddonfag N, Aontee A, Jaieau K, Kumsang P, Sripethdee C (2015) Packag Technol Sci 28:741Auras R, Harte B, Selke S (2004) J Appl Polym Sci 92:1790Sanchez-Garcia MD, Gimenez E, Lagaron JM (2008) Carbohyd Polym 71:235Sanchez-Garcia MD, Gimenez E, Lagaron JM (2007) J Plast Film Sheeting 23:133Lagaron JM (2011) Multifunctional and nanoreinforced polymers for food packaging. In: Multifunctional and nanoreinforced polymers for food packaging. Woodhead Publishing, Cambridge, p 

Hand-held echocardiography: added value in clinical cardiological assessment

BACKGROUND: The ultrasonic industry has recently produced echocardiographic Hand Held Devices (miniaturized, compact and battery-equipped echocardiographic systems). Their potential usefulness has been successfully assessed in a wide range of clinical conditions. The aim of the study was to verify if the routine use of a basic model of echocardiographic Hand Held Device (HHD) could be an important diagnostic tool during outpatient cardiologic consulting or in non-cardiologic hospital sections. METHODS: 87 consecutive patients were included in this study; they underwent routine physical examination, resting ECG and echocardiographic evaluation using a basic model of HHD performed by trained echocardiographists; the cardiologist, whenever possible, formulated a diagnosis. The percentage of subjects in whom the findings were judged reasonably adequate for final diagnostic and therapeutic conclusions was used to quantify the "conclusiveness" of HHD evaluation. Successively, all patients underwent a second echocardiographic evaluation, by an examiner with similar echocardiographic experience, performed using a Standard Echo Device (SED). The agreement between the first and the second echocardiographic exam was also assessed. RESULTS: Mean examination time was 6.7 ± 1.5 min. using HHD vs. 13.6 ± 2.4 min. using SED. The echocardiographic examination performed using HHD was considered satisfactory in 74/87 patients (85.1% conclusiveness). Among the 74 patients for whom the examination was conclusive, the diagnosis was concordant with that obtained with the SED examination in 62 cases (83.8% agreement). CONCLUSION: HHD may generally allow a reliable cardiologic basic evaluation of outpatient or subjects admitted to non-cardiologic sections, more specifically in particular subgroups of patients, with a gain in terms of time, shortening patient waiting lists and reducing healthy costs

Repetitive N-WASP–Binding Elements of the Enterohemorrhagic Escherichia coli Effector EspFU Synergistically Activate Actin Assembly

Enterohemorrhagic Escherichia coli (EHEC) generate F-actin–rich adhesion pedestals by delivering effector proteins into mammalian cells. These effectors include the translocated receptor Tir, along with EspFU, a protein that associates indirectly with Tir and contains multiple peptide repeats that stimulate actin polymerization. In vitro, the EspFU repeat region is capable of binding and activating recombinant derivatives of N-WASP, a host actin nucleation-promoting factor. In spite of the identification of these important bacterial and host factors, the underlying mechanisms of how EHEC so potently exploits the native actin assembly machinery have not been clearly defined. Here we show that Tir and EspFU are sufficient for actin pedestal formation in cultured cells. Experimental clustering of Tir-EspFU fusion proteins indicates that the central role of the cytoplasmic portion of Tir is to promote clustering of the repeat region of EspFU. Whereas clustering of a single EspFU repeat is sufficient to bind N-WASP and generate pedestals on cultured cells, multi-repeat EspFU derivatives promote actin assembly more efficiently. Moreover, the EspFU repeats activate a protein complex containing N-WASP and the actin-binding protein WIP in a synergistic fashion in vitro, further suggesting that the repeats cooperate to stimulate actin polymerization in vivo. One explanation for repeat synergy is that simultaneous engagement of multiple N-WASP molecules can enhance its ability to interact with the actin nucleating Arp2/3 complex. These findings define the minimal set of bacterial effectors required for pedestal formation and the elements within those effectors that contribute to actin assembly via N-WASP-Arp2/3–mediated signaling pathways

Development and validation of a population-based prediction scale for osteoporotic fracture in the region of Valencia, Spain: the ESOSVAL-R study

<p>Abstract</p> <p>Background</p> <p>Today, while there are effective drugs that reduce the risk of osteoporotic fracture, yet there are no broadly accepted criteria that can be used to estimate risks and decide who should receive treatment. One of the actual priorities of clinical research is to develop a set of simple and readily-available clinical data that can be used in routine clinical practice to identify patients at high risk of bone fracture, and to establish thresholds for therapeutic interventions. Such a tool would have high impact on healthcare policies. The main objective of the ESOSVAL-R is to develop a risk prediction scale of osteoporotic fracture in adult population using data from the Region of Valencia, Spain.</p> <p>Methods/Design</p> <p><it>Study design</it>: An observational, longitudinal, prospective cohort study, undertaken in the Region of Valencia, with an initial follow-up period of five years; <it>Subjects</it>: 14,500 men and women over the age of 50, residing in the Region and receiving healthcare from centers where the ABUCASIS electronic clinical records system is implanted; <it>Sources of data</it>: The ABUCASIS electronic clinical record system, complemented with hospital morbidity registers, hospital Accidents & Emergency records and the Regional Ministry of Health's mortality register; <it>Measurement of results</it>: Incident osteoporotic fracture (in the hip and/or major osteoporotic fracture) during the study's follow-up period. Independent variables include clinical data and complementary examinations; <it>Analysis</it>: 1) Descriptive analysis of the cohorts' baseline data; 2) Upon completion of the follow-up period, analysis of the strength of association between the risk factors and the incidence of osteoporotic fracture using Cox's proportional hazards model; 3) Development and validation of a model to predict risk of osteoporotic fracture; the validated model will serve to develop a simplified scale that can be used during routine clinical visits.</p> <p>Discussion</p> <p>The ESOSVAL-R study will establish a prediction scale for osteoporotic fracture in Spanish adult population. This scale not only will constitute a useful prognostic tool, but also it will allow identifying intervention thresholds to support treatment decision-making in the Valencia setting, based mainly on the information registered in the electronic clinical records.</p

Jet energy measurement with the ATLAS detector in proton-proton collisions at root s=7 TeV

The jet energy scale and its systematic uncertainty are determined for jets measured with the ATLAS detector at the LHC in proton-proton collision data at a centre-of-mass energy of √s = 7TeV corresponding to an integrated luminosity of 38 pb-1. Jets are reconstructed with the anti-kt algorithm with distance parameters R=0. 4 or R=0. 6. Jet energy and angle corrections are determined from Monte Carlo simulations to calibrate jets with transverse momenta pT≥20 GeV and pseudorapidities {pipe}η{pipe}<4. 5. The jet energy systematic uncertainty is estimated using the single isolated hadron response measured in situ and in test-beams, exploiting the transverse momentum balance between central and forward jets in events with dijet topologies and studying systematic variations in Monte Carlo simulations. The jet energy uncertainty is less than 2. 5 % in the central calorimeter region ({pipe}η{pipe}<0. 8) for jets with 60≤pT<800 GeV, and is maximally 14 % for pT<30 GeV in the most forward region 3. 2≤{pipe}η{pipe}<4. 5. The jet energy is validated for jet transverse momenta up to 1 TeV to the level of a few percent using several in situ techniques by comparing a well-known reference such as the recoiling photon pT, the sum of the transverse momenta of tracks associated to the jet, or a system of low-pT jets recoiling against a high-pT jet. More sophisticated jet calibration schemes are presented based on calorimeter cell energy density weighting or hadronic properties of jets, aiming for an improved jet energy resolution and a reduced flavour dependence of the jet response. The systematic uncertainty of the jet energy determined from a combination of in situ techniques is consistent with the one derived from single hadron response measurements over a wide kinematic range. The nominal corrections and uncertainties are derived for isolated jets in an inclusive sample of high-pT jets. Special cases such as event topologies with close-by jets, or selections of samples with an enhanced content of jets originating from light quarks, heavy quarks or gluons are also discussed and the corresponding uncertainties are determined. © 2013 CERN for the benefit of the ATLAS collaboration

A Situational Alignment Framework for PACS

This paper reports the outcomes of a study on an integrated situational alignment framework for picture archiving and communication systems (PACS) labeled as PISA. Following the design research cycle, complementary validation methods and pilot cases were used to assess the proposed framework and its operationalized survey. In this paper, the authors outline (a) the process of the framework’ development, (b) the validation process with its underlying iterative steps, (c) the outcomes of pilot cases, and (d) improvement opportunities to refine and further validate the PISA framework. Results of this study support empirical application of the framework to hospital enterprises in order to gain insights into their PACS maturity and alignment. We argue that the framework can be applied as a valuable tool for assessments, monitoring and benchmarking purposes and strategic PACS planning

Measurement of the inclusive and dijet cross-sections of b-jets in pp collisions at sqrt(s) = 7 TeV with the ATLAS detector

The inclusive and dijet production cross-sections have been measured for jets containing b-hadrons (b-jets) in proton-proton collisions at a centre-of-mass energy of sqrt(s) = 7 TeV, using the ATLAS detector at the LHC. The measurements use data corresponding to an integrated luminosity of 34 pb^-1. The b-jets are identified using either a lifetime-based method, where secondary decay vertices of b-hadrons in jets are reconstructed using information from the tracking detectors, or a muon-based method where the presence of a muon is used to identify semileptonic decays of b-hadrons inside jets. The inclusive b-jet cross-section is measured as a function of transverse momentum in the range 20 < pT < 400 GeV and rapidity in the range |y| < 2.1. The bbbar-dijet cross-section is measured as a function of the dijet invariant mass in the range 110 < m_jj < 760 GeV, the azimuthal angle difference between the two jets and the angular variable chi in two dijet mass regions. The results are compared with next-to-leading-order QCD predictions. Good agreement is observed between the measured cross-sections and the predictions obtained using POWHEG + Pythia. MC@NLO + Herwig shows good agreement with the measured bbbar-dijet cross-section. However, it does not reproduce the measured inclusive cross-section well, particularly for central b-jets with large transverse momenta.Comment: 10 pages plus author list (21 pages total), 8 figures, 1 table, final version published in European Physical Journal

Search for the neutral Higgs bosons of the minimal supersymmetric standard model in pp collisions at root s=7 TeV with the ATLAS detector

A search for neutral Higgs bosons of the Minimal Supersymmetric Standard Model (MSSM) is reported. The analysis is based on a sample of proton-proton collisions at a centre-of-mass energy of 7TeV recorded with the ATLAS detector at the Large Hadron Collider. The data were recorded in 2011 and correspond to an integrated luminosity of 4.7 fb-1 to 4.8 fb-1. Higgs boson decays into oppositely-charged muon or τ lepton pairs are considered for final states requiring either the presence or absence of b-jets. No statistically significant excess over the expected background is observed and exclusion limits at the 95% confidence level are derived. The exclusion limits are for the production cross-section of a generic neutral Higgs boson, φ, as a function of the Higgs boson mass and for h/A/H production in the MSSM as a function of the parameters mA and tan β in the mhmax scenario for mA in the range of 90GeV to 500 GeV. Copyright CERN

Observation of associated near-side and away-side long-range correlations in √sNN=5.02 TeV proton-lead collisions with the ATLAS detector

Two-particle correlations in relative azimuthal angle (Δϕ) and pseudorapidity (Δη) are measured in √sNN=5.02  TeV p+Pb collisions using the ATLAS detector at the LHC. The measurements are performed using approximately 1  μb-1 of data as a function of transverse momentum (pT) and the transverse energy (ΣETPb) summed over 3.1<η<4.9 in the direction of the Pb beam. The correlation function, constructed from charged particles, exhibits a long-range (2<|Δη|<5) “near-side” (Δϕ∼0) correlation that grows rapidly with increasing ΣETPb. A long-range “away-side” (Δϕ∼π) correlation, obtained by subtracting the expected contributions from recoiling dijets and other sources estimated using events with small ΣETPb, is found to match the near-side correlation in magnitude, shape (in Δη and Δϕ) and ΣETPb dependence. The resultant Δϕ correlation is approximately symmetric about π/2, and is consistent with a dominant cos⁡2Δϕ modulation for all ΣETPb ranges and particle pT