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Long-term live-cell microscopy with labeled nanobodies delivered by laser-induced photoporation

Nano Research 2020年第2期13卷 485-495页

作者： Jing Liu Tim Hebbrecht Toon Brans Eef Parthoens Saskia Lippens Chengnan Li Herlinde De Keersmaecker Winnok H.De Vos Stefaan C.De Smedt Rabah Boukherroub Jan Gettemans Ranhua Xiong Kevin Braeckmans Laboratory of General Biochemistry and physical Pharmacy Faculty of PharmacyGhent UniversityGhent B-9000Belgium Department of Biomolecular medicine Faculty of Medicine and Health SciencesGhent UniversityGhent B-9000Belgium VIB-UGent Center for Inflammation Research VIBGhent B-9000Belgium VIB bioimaging Core Ghent VIBGhent B-9000Belgium Department of Biomedical Molecular biology Ghent UniversityGhent B-9000Belgium Univ.Lille CNRSCentrale LilleISENUniv.ValenciennesUMR 8520-IEMNLille F-59000Franc Laboratory of Cell biology and Histology Department of Veterinary SciencesUniversity of Antwerp2020 AntwerpBelgium centre for Advanced Light Microscopy Ghent UniversityGhent B-9000Belgium

Fluorescence microscopy is the method of choice for studying intracellular ***,its success depends on *** of specific and stable markers.A prominent example of markers that are rapidly gaining interest are nanobodies(Nbs.-15 kDa),which can be functionalized with bright and photostable organic *** to their relatively small size and high specificity,Nbs offer great potential for high-quality long-term subcellular imaging,but suffer from the fact that they cannot spontaneously cross the plasma membrane of live *** have recently discovered that laser-induced photoporation is well suited to deliver extrinsic labels to living cells without compromising their *** a laser-based technology,it is readily compatible with light microscopy and the typical cell recipients used for *** by these promising initial results,we demonstrate here for the first time successful long-term imaging of specific subcellular structures with labeled nanobodies in living *** illustrate this using Nbs that target GFP/YFP-protein constructs accessible in the cytoplasm,actin-bundling protein Fascin,and the histone H2A/H2B *** an efficiency of more than 80%labeled cells and minimal toxicity(-2%),photoporation proved to be an excellent intracellular delivery method for ***-lapse microscopy revealed that cell division rate and migration remained unaffected,confirming excellent cell viability and *** conclude that laser-induced photoporation labeled Nbs can be easily delivered into living cells,laying the foundation for further development of a broad range of Nbs with intracellular targets as a toolbox for long-term live-cell microscopy.

关键词： laser-induced photoporation vapor nanobubble long-term microscopy imaging nanobody intracellular delivery living cell labeling

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The Cooperative Activities of CSLD2, CSLD3, and CSLD5 Are Required for Normal Arabidopsis Development

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Molecular Plant 2011年第6期4卷 1024-1037页

作者： Lan Yin Yves Verhertbruggen Ai Oikawa Chithra Manisseri Bernhard Knierim Lina Prak Jacob Kriiger Jensen Paul Knox Manfred Auer William G.T. Willatsa and Henrik Vibe Scheller Department of Plant biology and Biotechnology Faculty of Life Sciences University of Copenhagen DK-1871 Frederiksberg C Denmark Joint BioEnergy Institute Feedstocks Division 5885 Hollis St Emeryville CA 94608 USA Present address： Department of Arctic and Marine biology University of Tromso 9037 Tromse Norway physical Biosciences division Lawrence Berkeley National Laboratory Berkeley CA 94720 USA Present address： Molecular Phytopathology and Genetics University of Hamburg D-22609 Hamburg Germany f Joint BioEnergy Institute Technology Division 5885 Hollis St Emeryville CA 94608 USA Life Sciences division Lawrence Berkeley National Laboratory Berkeley CA 94720 USA Present address： Department of Plant biology Michigan State University East Lansing MI 48824 USA centre for Plant Sciences Faculty of Biological Sciences University of Leeds Leeds LS2 9JT UK Department of Plant and Microbial biology University of California Berkeley CA 94720 USA Joint BioEnergy Institute Technology Division 5885 Hoilis St Emeryville CA 94608 USA

Glycosyltransferases of the Cellulose Synthase Like D （CS/D） subfamily have been reported to be involved in tip growth and stem development in Arabidopsis. The csld2 and csld3 mutants are root hair defective and the csld5 mutant has reduced stem growth. In this study, we produced double and triple knockout mutants of CSLD2, CSLD3, and CSLD5. Unlike the single mutants and the csld2/csld3 double mutant, the csld2/csld5, csld3/csld5, and csld2/csld3/csld5 mutants were dwarfed and showed severely reduced viability. This demonstrates that the cooperative activities of CSLD2, CSLD3, and CSLD5 are required for normal Arabidopsis development, and that they are involved in important processes besides the specialized role in tip growth. The mutant phenotypes indicate that CSLD2 and CSLD3 have overlapping functions with CSLD5 in early plant development, whereas the CSLD2 and CSLD3 proteins are non-redundant. To determine the biochemical function of CSLD proteins, we used transient expression in tobacco leaves. Microsomes containing heterologously expressed CSLD5 transferred mannose from GDP-mannose onto endogenous acceptors. The same activity was detected when CSLD2 and CSLD3 were coexpressed but not when they were expressed separately. With monosaccharides as exogenous acceptors, microsomal preparations from CSLD5-expressing plants mediated the transfer of mannose from GDP-mannose onto mannose. These results were supported by immunodetection studies that showed reduced levels of a mannan epitope in the cell walls of stem interfascicular fibers and xylem vessels of the csld2/csld3/csld5 mutant.

关键词： Cellulose synthase like hemicelluloses mannan glycosyltransferases polysaccharide plant development.

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RRx-001 ameliorates inflammatory diseases by acting as a potent covalent NLRP3 inhibitor

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Cellular & Molecular Immunology 2021年第6期18卷 1425-1436页

作者： Yun Chen Hongbin He Bolong Lin Yun Chen Xianming Deng Wei Jiang Rongbin Zhou Hefei National Laboratory for physical Sciences at Microscale the CAS Key Laboratory of Innate Immunity and Chronic DiseaseSchool of Basic Medical SciencesDivision of Life Sciences and MedicineUniversity of Science and Technology of ChinaHefeiChina Department of Geriatrics Gerontology Institute of Anhui Provincethe First Affiliated Hospital of USTCDivision of Life Sciences and MedicineUniversity of Science and Technology of ChinaHefeiChina CAS centre for Excellence in Cell and Molecular biology University of Science and Technology of ChinaHefeiChina State Key Laboratory of Cellular Stress biology Innovation Center for Cell Signaling NetworkSchool of Life SciencesXiamen UniversityXiamenFujianChina

The NLRP3 inflammasome plays a crucial role in innate immune-mediated inflammation and contributes to the pathogenesis of multiple autoinflammatory,metabolic and neurodegenerative diseases,but medications targeting the NLRP3 inflammasome are not available for clinical ***-001 is a well-tolerated anticancer agent currently being investigated in phase III clinical trials,but its effects on inflammatory diseases are not ***,we show that RRx-001 is a highly selective and potent NLRP3 inhibitor that has strong beneficial effects on NLRP3-driven inflammatory ***-001 inhibits the activation of the canonical,noncanonical,and alternative NLRP3 inflammasomes but not the AIM2,NLRC4 or Pyrin ***,RRx-001 covalently binds to cysteine 409 of NLRP3 via its bromoacetyl group and therefore blocks the NLRP3-NEK7 interaction,which is critical for the assembly and activation of the NLRP3 *** importantly,RRx-001 treatment attenuates the symptoms of lipopolysaccharide(LPS)-induced systemic inflammation,dextran sulfate sodium(DSS)-induced colitis and experimental autoimmune encephalomyelitis(EAE)in ***,our study identifies RRx-001 as a new potential therapeutic agent for NLRP3-driven diseases.

关键词： RRx-001 NLRP3 inflammasome inflammatory diseases

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Phase-Controlled 1T Transition-Metal Dichalcogenide-Based Multidimensional Hybrid Nanostructures

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CCS Chemistry 2021年第11期3卷 58-68页

作者： Hou-Ming Xu Chao Gu Xiao-Long Zhang Lei Shi Qiang Gao Shaojin Hu Shi-Kui Han Xiao Zheng Min-Rui Gao Shu-Hong Yu Key Laboratory of Advanced Catalytic Materials and Reaction Engineering School of Chemistry and Chemical EngineeringHefei University of TechnologyHefei 230009 Hefei National Laboratory for physical Sciences at the Microscale Division of Nanomaterials and ChemistryDepartment of ChemistryCAS Centre for Excellence in NanoscienceHefei Science Centre of CASHefei Comprehensive National Science CentreInstitute of EnergyUniversity of Science and Technology of ChinaHefei 230026 Hefei National Laboratory for physical Sciences at the Microscale Division of Theoretical and Computational SciencesCAS Centre for Excellence and Synergetic Innovation Centre in Quantum Information and Quantum PhysicsUniversity of Science and Technology of ChinaHefei 230026

Metallic-phase transition-metal dichalcogenides(TMDCs)exhibit unusual physicochemical properties compared with their semiconducting ***,they are thermodynamically unstable to access and it is even more challenging to construct their metastable-phase ***,we demonstrate a general solution protocol for phase-controlled synthesis of distorted octahedral 1T WS2-based(1T structure denotes an octahedral coordination for W atom)multidimensional hybrid nanostructures from two-dimensional(2D),one-dimensional(1D),and zero-dimensional(0D)*** is realized by tuning the reactivity of tungsten precursor and the interaction between crystal surface and *** a conceptual study on crystal phase-and dimensionality-dependent applications,we find that the three-dimensional(3D)hierarchical architectures achieved,comprising 1T WS2 and 2D Ni3S4,are very active and stable for catalyzing hydrogen *** results open up a new way to rationally design phase-controlled nanostructures with increased complexity and more elaborate functionalities.

关键词： wet-chemical synthesis phase transitionmetal dichalcogenides hybrid nanostructures hydrogen evolution

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Phosphorene: Current status, challenges and opportunities

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Frontiers of Chemical Science and Engineering 2019年第2期13卷 296-309页

作者： Anandarup Goswami Manoj B.Gawande division of Chemistry Department of Sciences and HumanitiesVignan's Foundation for ScienceTechnology and Research (VFSTR)VadlamudiGuntur-522213Andhra PradeshIndia Regional centre of Advanced Technologies and Materials Department of Physical ChemistryFaculty of SciencePalacky University OlomoucSlechtitelu 27783 71 OlomoucCzech Republic

The field of 2-dimensional (2D) materials has witnessed a sharp growth since its inception and can majorly be attributed to the substantial technical and scientific developments, leading to significant improvements in their syntheses, characterization and applications. In the list of 2D materials, the relatively newer addition is phosphorene, which ideally consists of a single layer of black phosphorous. Keeping in mind the past, and ongoing research activities, this short account offers a brief overview of the present status and the associated challenges in the field of phosphorene-related research, with special emphasis on their syntheses, properties, applications and future opportunities.

关键词： phosphorene black phosphorous anisotropy single layer thermoelectric chemical vapor deposition catalysis battery supercapacitor

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Mill tailings based composites as paste backfill in mines of U-bearing dolomitic limestone ore

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Journal of Rock Mechanics and Geotechnical Engineering 2018年第2期10卷 310-322页

作者： Sandeep Panchal Debasis Deb T.Sreenivas Department of Mining Engineering Indian Institute of Technology Mineral Processing division Bhabha Atomic Research Centre

This paper elaborates on the development of paste backfill using mill tailings generated during the processing of a uranium ore deposit hosted in dolomitic limestone. The tailings have been characterized in terms of the physical, chemical and mineralogical properties. Time-dependent rheological behaviors and geotechnical properties of cemented paste backfill(CPB) are also determined. The studies show that the mill tailing has the potential to form paste and the CPB has adequate strength to provide support to mine pillars, roofs, and walls.

关键词： Mining engineering Uranium ore deposit Tailings Cemented paste backfill(CPB) Rheology Compressive strength

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3D assessment of intervertebral disc degeneration in zebrafish identifies changes in bone density that prime disc disease

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Bone Research 2021年第4期9卷 521-536页

作者： Erika Kague Francesco Turci Elis Newman Yushi Yang Kate Robson Brown Mona S.Aglan Ghada A.Otaify Sarnia A.Temtamy Victor L.Ruiz-Perez Stephen Cross C.Patrick Royall P.Eckhard Witten Chrissy L.Hammond School of Physiology Pharmacology and NeuroscienceBiomedical SciencesUniversity of BristolBristolUK School of Physics HH Wills Physics LaboratoryUniversity of BristolBristolUK centre for Nanoscience and Quantum Information University of BristolBristolUK Bristol centre for Functional Nanomaterials University of BristolBristolUK Department of Anthropology and Archaeology University of BristolBristolUK Department of Mechanical Engineering University of BristolBristolUK Clinical Genetics Department Human Genetics and Genome Research DivisionCenter of Excellence for Human GeneticsNational Research CentreCairoEgypt lnstituto de Investigaciones Biomedicas de Madridand Ciber de Enfermedades Raras(CIBERER)MadridSpain Wolfson bioimaging Facility Biomedical SciencesUniversity of BristolBristolUK School of Chemistry University of BristolBristolUK Evolutionary Developmental biology Department of BiologyGhent UniversityGhentBelgium

Back pain is a common condition with a high social impact and represents a global health *** disc disease(IVDD)is one of the major causes of back pain;no therapeutics are currently available to reverse this *** impact of bone mineral density(BMD)on IVDD has been controversial,with some studies suggesting osteoporosis as causative for IVDD and others suggesting it as protective for *** studies to evaluate the influence of genetic components of BMD in IVDD could highlight opportunities for drug development and *** taking a holistic 3D approach,we established an aging zebrafish model for spontaneous *** BMD in aging,detected by automated computational analysis,is caused by bone deformities at the ***,aged zebrafish spines showed changes in bone morphology,microstructure,mineral heterogeneity,and increased fragility that resembled *** of the discs recapitulated IVDD symptoms found in humans:the intervertebral ligament(equivalent to the annulus fibrosus)showed disorganized collagen fibers and herniation,while the disc center(nucleus pulposus equivalent)showed dehydration and cellular *** manipulated BMD in young zebrafish by mutating sp7 and cathepsin K,leading to low and high BMD,***,we detected IVDD in both groups,demonstrating that low BMD does not protect against IVDD,and we found a strong correlation between high BMD and *** learning was applied to high-resolution synchrotron\iCJ image data to analyze osteocyte 3D lacunar distribution and morphology,revealing a role of sp7 in controlling the osteocyte lacunar 3D *** findings suggest potential avenues through which bone quality can be targeted to identify beneficial therapeutics for IVDD.

关键词： intervertebral degeneration protective

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Laser-Heated Diamond-Anvil Cell (LHDAC) in Materials Science Research

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Journal of Materials Science & Technology 2003年第6期19卷 518-525页

作者： N.V.Chandra Shekar P.Ch.Sahu K.Govinda Rajan Materials Science division Indira Gandhi Centre for Atomic Research Kalpakkam 603 102 Tamil Nadu IndiaMaterials Science Division Indira Gandhi Centre for Atomic Research Kalpakkam 603 102 Tamil Nadu IndiaMaterials Science Division Indira Gandhi Centre for Atomic Research Kalpakkam 603 102 Tamil Nadu India

Laser-heated diamond-anvil cell (LHDAC) is emerging as the most suitable, economical and versatile tool for the measurement of a large spectrum of physical properties of materials under extreme pressure and temperature conditions. In this review, the recent developments in the instrumentation, pressure and temperature measurement techniques, results of experimental investigations from the literature were discussed. Also, the future scope of the technique in various avenues of science was explored.

关键词： Laser heating Diamond-anvil cell High pressure-high temperature Materials synthesis,Melting phenomena Phase equilibria

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Room-temperature third-order nonlinear Hall effect in Weyl semimetal TalrTe4

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National Science Review 2022年第12期9卷 177-184页

作者： Cong Wang Rui-Chun Xiao Huiying Liu Zhaowei Zhang Shen Lai Chao Zhu Hongbing Cai Naizhou Wang Shengyao Chen Ya Deng Zheng Liu Shengyuan A.Yang Wei-Bo Gao College of Mathematics and Physics Beijing University of Chemical Technology division of Physics and Applied Physics School of Physical and Mathematical Sciences Nanyang Technological University Institute of physical Science and Information Technology Anhui University Research Laboratory for Quantum Materials Singapore University of Technology and Design School of Materials Science and Engineering Nanyang Technological University CAS Center for Excellence in Nanoscience National Center for Nanoscience and Technology The Photonics Institute and centre for Disruptive Photonic Technologies Nanyang Technological University

The second-order nonlinear Hall effect observed in the time-revers al symmetric system has not only shown abundant physical content,but also exhibited potential application ***,a third-order nonlinear Hall effect has been observed in MoTe2and ***,few-layer MoTe2and WTe2are usually unstable in air and the observed third-order nonlinear Hall effect can be measured only at low temperature,which hinders further investigation as well as potential ***,exploring new air-stable material systems with a sizable third-order nonlinear Hall effect at room temperature is an urgent ***,in type-Ⅱ Weyl semimetal TaIrTe4,we observed a pronounced third-order nonlinear Hall effect,which can exist at room temperature and remain stable for *** third-order nonlinear Hall effect is connected to the Berry-connection polarizability tensor instead of the Berry *** possible me chanism of the observation of the third-order nonlinear Hall effe ct in TaIrTe4at room temperature has been *** findings will open an avenue towards exploring room-temperature nonlinear devices in new quantum materials.

关键词： topological material third-order nonlinear Hall effect room temperature Berry connection polarizability tensor

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Arabidopsis Mutants and the Network of M icrotu bu le-Associated Functions

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Molecular Plant 2008年第6期1卷 888-898页

作者： Henrik Buschmann Clive W. Lloyd Department of Cell and Developmental biology John Innes Centre 1 Colney Lane Norwich NR4 7UH UK

In early eukaryotes, the microtubule system was engaged in mitosis, intracellular transport, and flagellumbased motility. In the plant lineage, the evolution of a multicellular body involved the conservation of some core functions, the loss of others, and the elaboration of new microtubule functions associated with the multicellular plant habit. This diversification is reflected by the presence of both conserved （animal/fungi-like） and novel （plant-like） sequences encoding microtubule-related functions in the Arabidopsis genome. The collection of microtubule mutants has grown rapidly over recent years. These mutants present a wide range of phenotypes, consistent with the hypothesis of a functional diversification of the microtubule system. In this review, we focus on mutant analysis and, in particular, discuss double mutant analysis as a valuable tool for pinpointing pathways of gene function. A future challenge will be to define the complete network of genetic and physical interactions of microtubule function in plants. In addition to reviewing recent progress in the functional analysis of the ＇MAPome＇, we present an online database of Arabidopsis mutants impaired in microtubule functions.

关键词： cell division cell morphogenesis cytoskeleton genetics development Arabidopsis.

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