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Three-dimensional visualization of planta clathrin-coated vesicles at ultrastructural resolution

Molecular Plant 2022年第10期15卷 1533-1542页

作者： Alexander Johnson Walter A.Kaufmann Christoph Sommer Tommaso Costanzo Dana A.Dahhan sebastian y.bednarek Jiri Friml Institute of Science and Technology Austria(ISTA) 3400KlosterneuburgAustria UW-Madison Department of Biochemistry433 Babcock Dr.MadisonWI 53706USA

Biological systems are the sum of their dynamic three-dimensional(3D)***,it is critical to study biological structures in 3D and at high resolution to gain insights into their physiological *** microscopy of metal replicas of unroofed cells and isolated organelles has been a key technique to visualize intracellular structures at nanometer ***,many of these methods require specialized equipment and personnel to complete ***,we present novel accessible methods to analyze biological structures in unroofed cells and biochemically isolated organelles in 3D and at nanometer resolution,focusing on Arabidopsis clathrin-coated vesicles(CCVs).While CCVs are essential trafficking organelles,their detailed structural information is lacking due to their poor preservation when observed via classical electron microscopy protocols ***,we establish a method to visualize CCVs in unroofed cells using scanning transmission electron microscopy tomography,providing sufficient resolution to define the clathrin coat ***,the samples are prepared directly on electron microscopy grids,removing the requirement to use extremely corrosive acids,thereby enabling the use of this method in any electron microscopy ***,we demonstrate that this standardized sample preparation allows the direct comparison of isolated CCV samples with those visualized in ***,to facilitate the high-throughput and robust screening of metal replicated samples,we provide a deep learning analysis method to screen the“pseudo 3D”morphologies of CCVs imaged with 2D ***,our work establishes accessible ways to examine the 3D structure of biological samples and provide novel insights into the structure of plant CCVs.

关键词： Arabidopsis thaliana clathrin-coated vesicles STEM tomography electron microscopy clathrin-mediated endocytosis

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Differential regulation of clathrin and its adaptor proteins, AP-2 and the TPLATE complex, during their membrane recruitment in Arabidopsis

Differential regulation of clathrin and its adaptor proteins...

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2016年全国植物生物学大会

作者： Chao Wang Tianwei Hu Xu yan Tingting Meng yutong Wang Qingmei Wang Xiaoyue Zhang ying Gu Clara Sánchez-Rodríguez Astrid Gadeyne Jinxing Lin Staffan Persson Dani?l Van Damme Chuanyou Li sebastian y.bednarek Jianwei Pan College of Chemistry and Life Sciences Zhejiang Normal University Institute of Genetics and Developmental Biology Chinese Academy of Sciences Department of Biochemistry and Molecular Biology Pennsylvania State UniversityUniversity Park Department of Biology Institute of Agricultural SciencesETH Zurich Department of Plant Systems Biology VIBDepartment of Plant Biotechnology and BioinformaticsGhent University College of Biological Sciences and Biotechnology Beijing Forestry University

In plants,clathrin-mediated endocytosis(CME) is dependent on the function of clathrin and its accessory heterooligomeric adaptor protein complexes,Adaptor Protein 2(AP-2) and the TPLATE complex(TPC),and is negatively regulated by the hormones auxin and salicylic acid(SA). The details for how clathrin and its adaptor complexes are recruited to the plasma membrane(PM) to regulate CME are however poorly understood. We found that SA and the pharmacological CME inhibitor tyrphostin A23(Tyr A23) reduce the membrane association of clathrin and AP-2,but not that of the TPC,whereas auxin solely affected clathrin membrane association,in Arabidopsis thaliana. Genetic and pharmacological experiments revealed that loss of AP2μ or AP2σ partially affected the membrane association of other AP-2 subunits and that the AP-2 subunit AP2σ,but not AP2μ,was required for SA-and Tyr A23-dependent inhibition of CME. Furthermore,we show that although AP-2 and the TPC are both required for the PM recruitment of clathrin in wild-type cells,the TPC is necessary for clathrin PM association in AP-2 deficient cells. These results indicate that developmental signals may differentially modulate the membrane recruitment of clathrin and its core accessory complexes to regulate the process of CME in plant cells.

关键词： Adaptor Protein 2 auxin clathrin endocytosis salicylic acid TPLATE complex tyrphostin A23 Arabidopsis

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Rainfall Sensitivity to Microphysics and Planetary Boundary Layer Parameterizations in Convection-Permitting Simulations over Northwestern South America

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Journal of Meteorological Research 2024年第4期38卷 805-825页

作者： K.Santiago HERNANDEZ sebastian GOMEZ-RIOS Juan J.HENAO Vanessa ROBLEDO Alvaro RAMiREZ-CARDONA Angela M.RENDON Geolimna Escuela AmbientalFacultad de IngenieriaUniversidad de AntioquiaMedellin 050010Colombia Grupo de Ingenieria y Gestion Ambiental(GIGA) Escuela AmbientalFacultad de IngenieriaUniversidad de AntioquiaMedellin 050010Colombia Division of Atmospheric Sciences Desert Research InstituteRenoNV 89512USA

Convection-permitting modeling allows us to understand mechanisms that influence rainfall in specific ***,microphysics parameterization(MP) and planetary boundary layer(PBL) schemes remain an important source of uncertainty,affecting rainfall intensity,occurrence,duration,and ***,we study the sensitivity of rainfall to three MP [Weather Research and Forecasting(WRF) Single-Moment 6-class(WSM6),Thompson,and Morrison] and two PBL [the yonsei University(ySU) and Mellor–yamada Nakanishi Niino(MyNN)] schemes with a convection-permitting resolution(4 km) over northwestern South America(NWSA).Simulations were performed by using the WRF model and the results were evaluated against soundings,rain gauges,and satellite data,considering the spatio-temporal variability of rainfall over diverse regions prone to deep convection in *** and PBL schemes largely influenced simulated rainfall,with better results for the less computationally expensive WSM6 MP and ySU PBL *** rain gauges and satellite estimates,simulations with Morrison MP overestimated rainfall,especially westward of the Andes,whereas the MyNN PBL underestimated precipitation in the Amazon–Savannas *** found that the uncertainty in the rainfall representation is highly dependent on the region,with a higher influence of MP in the Colombian Pacific and PBL in the Amazon–Savannas *** analyzing rainfall-related processes,the selection of both MP and PBL parameterizations exerted a large influence on the simulated lower tropospheric moisture flux and moisture *** schemes significantly influenced the downward shortwave radiation,with MyNN simulating a greater amount of low clouds,which decreased the radiation ***,latent heat fluxes were greater for ySU,favoring moist convection and *** schemes had a marked impact on vertical ***,Morrison MP showed stronger convection and higher precipitation rates,which is assoc

关键词： sensitivity microphysics planetary boundary layer(PBL) Weather Research and Forecasting(WRF) processes

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Optical magnetic field enhancement using ultrafast azimuthally polarized laser beams and tailored metallic nanoantennas

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Photonics Research 2024年第5期12卷 1078-1092页

作者： RODRIGO MARTIN-HERNANDEZ LORENZ GRUNEWALD LUIS SANCHEZ-TEJERINA LUIS PLAJA ENRIQUECONEJERO JARQUE CARLOS HERNANDEZ-GARCIA sebastian MAI Grupo de Investigacion en Aplicaciones del Laser y Fotonica Departamento de Fisica AplicadaUniversidad de SalamancaE-37008 SalamancaSpain Unidad de Excelencia en Luz y Materia Estructuradas(LUMES) Universidad de SalamancaE-37008 SalamancaSpain Institute of Theoretical Chemistry Faculty of ChemistryUniversity of Vienna1090 ViennaAustria Vienna Doctoral School in Chemistry(DoSChem) Faculty of ChemistryUniversity of Vienna1090 ViennaAustria Departamento de Electricidad y Electrónica Universidad de Valladolid47011 ValladolidSpain

*** *** thank Valentina Shumakova,Elizaveta Gangrskaia,Alessandra Bellissimo,Audrius Pugzlys,and Andrius Baltu?ka for fruitful discussions and computational support by the Vienna Scientific Cluster(VSC 4).The authors thankfully acknowledge the computer resources at MareNostrum and SCAyLE,and the technical support provided by Barcelona Supercomputing Center(RES-FI-2022-3-0041).R.M.-H.,L.S.-T.,L.P.,E.C.J.,and C.H.-*** funding from the European Research Council(ERC)under the European Union’s Horizon 2020 research and innovation program(Grant Agreement No.851201)and support from Ministerio de Ciencia e Innovación(PID2022-142340NB-I00,PID2019-106910GB-I00).This research was funded in whole or in part by the Austrian Science Fund(FWF)(10.55776/ZK91).For open access purposes,the author has applied a CC By public copyright license to any author accepted manuscript version arising from this submission.

关键词： metallic polarized discussions

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Regulation of Pathogen-Triggered Tryptophan Metabolism in Arabidopsis thaliana by MyB Transcription Factors and Indole Glucosinolate Conversion Products

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Molecular Plant 2016年第5期9卷 682-695页

作者： Henning Frerigmann Mariola Pislewska-bednarek Andrea Sanchez-Vallet Antonio Molina Erich Glawischnig Tamara Gigolashvili Pawel bednarek Botanical Institute and Cluster of Excellence on Plant Sciences(CEPLAS) University of Cologne BioCenter Zulpicher Straβe 47b 50674 Cologne Germany Institute of Bioorganic Chemistry Polish Academy of Sciences Noskowskiego12/14 61-704 Pozna(n) Poland Centro de Biotecnologiay Genomica de Plantas Unlversidad Politecnica de Madrid Campus Montegancedo Pozuelo de Alarcon 28223 Madrid Spain Lehrstuhl for Genetik Technische Universitat Munchen Emil-Ramann-Str. 8 85354 Freising Germany ETH Zurich Department of Environmental Systems Science GHN H41 Universitatsstrasse 16 8092 Zurich Switzerland

MyB34, MyB51, and MyB122 transcription factors are known as decisive regulators of indolic glucosinolate （IG） biosynthesis with a strong impact on expression of genes encoding CyP79B2 and CyP79B3 enzymes that redundantly convert tryptophan to indole-3-acetaldoxime （IAOx）. This intermediate represents a branching point for IG biosynthesis, and pathways leading to camalexin and indole-carboxylic acids （ICA）. Here we investigate how these MyBs affect the pathogen-triggered Trp metabolism. Our experiments indicated that these three MyBs affect not only IG production but also constitutive biosynthesis of other IAOx- derived metabolites. Strikingly, the PENETRATION 2 （PEN2）-dependent IG-metabolism products, which are absent in myb34/51/122 and pen2 mutants, were indispensable for full flg22-mediated induction of other IAOx-dedved compounds. However, germ induction and accumulation of ICAs and camalexin upon path- ogen infection was not compromised in myb34/51/122 plants, despite strongly reduced IG levels. Hence, in comparison with cyp79B2/B3, which lacks all IAOx-derived metabolites, we found myb34/51/122 an ideal tool to analyze IG contribution to resistance against the necrotrophic fungal pathogen Plectosphaerella cucumerina. The susceptibility of myb34/51/122 was similar to that of pen2, but much lower than susceptibility of cyp79B2/B3, indicating that MyB34/51/122 contribute to resistance toward P. cucumerina exclu- sively through IG biosynthesis, and that PEN2 is the main leaf myrosinase activating IGs in response to microbial pathogens.

关键词： indolic glucosinate P/ectosphaere//a cucumerina MAMP-triggered immunity Trp metabolism indole-3-carboxylic acids camalexin

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Indentation Size Effect inβCuAlBe and Cu-2Be Alloys

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Acta Metallurgica Sinica(English Letters) 2021年第12期34卷 1669-1678页

作者： Susana Montecinos sebastian Tognana Walter Salgueiro IFIMAT Facultad de Ciencias ExactasUniversidad Nacional del Centro de La Provincia de Buenos AiresPinto 3997000 TandilArgentina Consejo Nacional de Investigaciones Científicas y Técnicas CONICET Av.Rivadavia 1917C1033AAJ Buenos AiresArgentina CIFICEN(UNCPBA-CICPBA-CONICET) Pinto 3997000 TandilArgentina Comisión de Investigaciones Científicas de la Provincia de Bs.As Calle 526 entre 10 y 111900 La PlataArgentina

The indentation size effect(ISE)was studied by instrumented nanoindentation and Vickers hardness measurements on polycrystalline copper,βCuAlBe pseudoelastic and age-hardenable CuBe *** in the load-depth curves at different loads would suggest a change in the behavior of the materials as the load *** was estimated at different loads taking into account the pile-up in each case,which was estimated from the images of the topography obtained by atomic force microscopy(AFM).The effect of the microstructure of Cu-2Be(wt%)samples on the ISE was also *** each copper alloy,the Nix and Gao model was applied to the hardness-depth curves,and significant differences were found for the parameters *** is in the constant nanohardness regime for depths below 500 nm,while microhardness exhibits the ISE *** behavior of the nanohardness and the plastic strain with the depth is almost *** presents the lowest values of plastic strains due to the pseudoelastic effect,while Cu presents an almost constant value around 8%,which corresponds to the highest plastic strain obtained.

关键词： Hardness Pseudoelasticity Copper alloys Plasticity Indentation size effect

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Coupling between fluids and rock deformation in the continental crust:Preface

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Geoscience Frontiers 2019年第6期10卷 2085-2092页

作者： Silvia Japas sebastian Oriolo Vinod Samuel Instituto de Geociencias Básicas Aplicadas y Ambientales(IGeBA)CONICETUniversidad de Buenos AiresCiudad UniversitariaBuenos AiresArgentina Department of Earth System Sciences Yonsei UniversitySeoulRepublic of Korea

*** Fluids are essential and widespread components of the Earth *** regulate the main process of mass and energy transport,conditioning the geochemical and geophysical evolution of the crust(Bredehoeft and Norton.1990)and,consequently,its properties and mechanical *** coupling between fluids,rock deformation,and heat are crucial engines for plate tectonics,playing a leading role in the rheology and evolution of the lithosphere(*** et al.,2000;Kennedy and van Soest,2007;Thompson,2010;Miller.2013;yardley and Bodnar,2014:Hirauchi et al.,2016:Menzies et al.,2016;Plümper et al.,2017;Vizán et al.,2017).

关键词： rock crust continental

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Clathrin-mediated membrane trafficking in Arabidopsis

Clathrin-mediated membrane trafficking in Arabidopsis

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2013全国植物生物学大会

作者： WANG Chao yAN Xu CHEN Qian JIANG Nan FU Wei MA Bojun LIU Jianzhong LI Chuanyou sebastian y.bednarek PAN Jianwei College of Chemistry and Life Sciences Zhejiang Normal University State Key Laboratory of Plant Genomics National Center for Plant Gene Research(Beijing)Institute of Genetics and Developmental BiologyChinese Academy of Sciences Department of Biochemistry University of WisconsinMadisonWisconsin 53706

Plant clathrin-mediatedmembrane trafficking is involved in many developmental processes as well as in responses to environmental *** studies have shown that clathrin-mediated endocytosis of the plasma membrane(PM)auxin transporter PIN-FORMED 1(PIN1)is regulated by the extracellular auxin receptor,AUXIN-BINDING PROTEIN 1 (ABP1).However,the mechanisms by which ABP1 and other factors regulate clathrin-mediated trafficking are poorly ***,we applied a genetic strategy and time-resolved imaging to dissect the role of clathrin light chains(CLCs)and ABP1 in auxin regulation of clathrin-mediated trafficking in *** was found to differentially regulate the PM-and trans-Golgi network/early endosome(TGN/EE)-association of CLCs and heavy chains (CHCs)in an ABP1-dependent but TIR1/AFB-independent *** of CLC2 and CLC3 affected CHC membrane-association,decreased both internalization and intracellular trafficking of PM proteins,and impaired auxin-regulated *** with these results, basipetal auxin transport,auxin sensitivity and distribution,and root gravitropism were also found to be dramatically altered in clc2 clc3 double mutants,resulting in pleiotropic defects in plant *** results suggest that CLCs are key regulators in clathrin-mediated trafficking downstream of ABP 1-mediated signaling and thus play a critical role in membrane trafficking from the TGN/EE and PM during plant development.

关键词： auxin clathrin light chains clathrin-mediated trafficking Arabidopsis thaliana

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A resistant method for landmark-based analysis of individual asymmetry in two dimensions

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Frontiers of Electrical and Electronic Engineering in China 2016年第4期4卷 270-282页

作者： sebastian Torcida Paula Gonzalez Federico Lotto Dpto. Matematica-Exactas UNCPBA. Tandil 7000 Argentina. Fac. de Ciencias Naturales y Museo UNLP-CONICET. La Plata 1900 Argentina Instituto de Veterinaria Ing. Fernando N. Dulout (IGEVET) Facultad de Cs. Veterinarias UNLP-CONICET. La Plata 1900Argentina

Symmetry of biological structures can be thought as the repetition of their parts in different positions and orientations. Asymmetry analyses, therefore, focuses on identifying and measuring the location and extent of symmetry departures in such structures. In the context of geometric morphometrics, a key step when studying morphological variation is the estimation of the symmetric shape. The standard procedure uses the least-squares Procrustes superimposition, which by averaging shape differences often underestimates the symmetry departures thus leading to an inaccurate description of the asymmetry pattern. Moreover, the corresponding asymmetry values are neither geometrically intuitive nor visually perceivable. Methods： In this work, a resistant method for landmark-based asymmetry analysis of individual bilateral symmetric structures in 2D is introduced. A geometrical derivation of this new approach is offered, while its advantages in comparison with the standard method are examined and discussed through a few illustrative examples. Results： Experimental tests on both artificial and real data show that asymmetry is more effectively measured by using the resistant method because the underlying symmetric shape is better estimated. Therefore, the most asymmetric （respectively symmetric） landmarks are better determined through their large （respectively small） residuals. The percentage of asymmetry that is accounted for by each landmark is an additional revealing measure the new method offers which agrees with the displayed results while helping in their biological interpretation. Conclusions： The resistant method is a useful exploratory tool for analyzing shape asymmetry in 2D, and it might be the preferable method whenever a non homogeneous deformation of bilateral symmetric structures is possible. By offering a more detailed and rather exhaustive explanation of the asymmetry pattern, this new approach will hopefully contribute to improve the quality of biological or

关键词： resistant procrustes method shape asymmetry matching and object symmetry landmarks

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TopSTO:a 115-line code for topology optimization of structures under stationary stochastic dynamic loading

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Earthquake Engineering and Engineering Vibration 2023年第4期22卷 1081-1100页

作者： sebastian Pozo Fernando Gomez Thomas Golecki Juan Carrion Billie F.Spencer Jr. Civil and Environmental Engineering University of Illinois at Urbana-ChampaignUrbana 61801ILUSA College of Science and Engineering Universidad San Francisco de QuitoDiego de Robles y Vía InteroceanicaQuitoEcuador Skidmore Owings&MerrillLLPChicago 60604ILUSA Department of Civil Engineering University of CuencaCuencaEcuador

The use of topology optimization in structural design under dynamic excitation is becoming more prevalent in the *** many such applications utilize frequency or time domain formulations,relatively few consider stochastic dynamic *** paper presents an efficient and compact code called TopSTO for structural topology optimization considering stationary stochastic dynamic loading using a method derived from random vibration *** theory,described in conjunction with the implementation in the provided code,is illustrated for a seismically excited *** work demonstrates the efficiency of the approach in terms of both the computational resources and minimal amount of code *** code is intended to serve as a baseline for understanding the theory and implementation of this topology optimization approach and as a foundation for additional applications and developments.

关键词： topology optimization stochastic dynamics matlab python

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