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Glucose transporter 4 can be inserted in the membrane without exposing its catalytic site for photolabeling from the medium

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Science China(Life Sciences) 2007年第2期50卷 147-154页

作者： Manabu ISHIKI philip j bilan Programme in Cell Biology Hospital for Sick Children Toronto Canada M5G 1×8

Insulin stimulates the production of PI(3,4,5)P3 in muscle cells, and this is required to stimulate GLUT4 fusion with the plasma membrane. Introduction of exogenous PI(3,4,5)P3 to muscle cells recapitulates insulin's effects on GLUT4 fusion with the plasma membrane, but not glucose uptake. This study aims to explore the mechanism behind this difference. In L6-GLUT4myc muscle cells, the availability of the GLUT4 intracellular C-terminus and extracellular myc epitopes for immunoreactivity on plasma membrane lawns was detected with the corresponding antibody. The availability of the active site of GLUT4 from extracellular medium was assessed by affinity photolabeling with the cell impermeant compound Bio-LC-ATB-BMPA. 100nmol/L insulin and 10μmol/L PI(3,4,5)P3 caused myc signal gain on the plasma membrane lawns by 1.64-fold and 1.58-fold over basal, respectively. Insulin, but not PI(3,4,5)P3, increased photolabeling of GLUT4 and immunolabeling with C-terminus antibody by 2.47-fold and 2.04-fold over basal, respectively. Upon insulin stimulation, the C-terminus signal gain was greater than myc signal gain (2.04-fold vs. 1.64-fold over basal, respectively) in plasma membrane lawns. These results indicate that (i) PI(3,4,5)P3 does not make the active site of GLUT4 available from the extracellular surface despite causing GLUT4 fusion with the plasma membrane; (ii) the availability of the active site of GLUT4 from the extracellular medium and availability of the C-terminus from the cytosolic site are correlated; (iii) in addition to stimulating GLUT4 translocation, insulin stimulation displaces a protein which masks the GLUT4 C-terminus. We propose that a protein which masks the C-terminus also prevents the active site from being available for photolabelling and possibly glucose uptake after treatment with PI(3,4,5)P3.

关键词： glucose transport 4 (GLUT4) insulin phosphatidylinositol 3 4 5-trisphosphate (PI(3 4 5)P3) photolabel

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Insulin sensitivity and inhibition by forskolin,dipyridamole and pentobarbital of glucose transport in three L6 muscle cell lines

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Science China(Life Sciences) 2007年第6期50卷 739-747页

作者： philip j bilan Michiko HAYASHI Programme in Cell Biology Hospital for Sick Children Toronto Canada M5G 1X8

L6 skeletal muscle myoblasts stably overexpressing glucose transporter GLUT1 or GLUT4 with exofa- cial myc-epitope tags were characterized for their response to insulin. In clonally selected cultures, 2-deoxyglucose uptake into L6-GLUT1myc myoblasts and myotubes was linear within the time of study. In L6-GLUT1myc and L6-GLUT4myc myoblasts, 100 nmol/L insulin treatment increased the GLUT1 content of the plasma membrane by 1.58±0.01 fold and the GLUT4 content 1.96±0.11 fold, as well as the 2-deoxyglucose uptake 1.53±0.09 and 1.86±0.17 fold respectively, all by a wortmannin-inhibitable manner. The phosphorylation of Akt in these two cell lines was increased by insulin. L6-GLUT1myc myoblasts showed a dose-dependent stimulation of glucose uptake by insulin, with unaltered sensitiv- ity and maximal responsiveness compared with wild type cells. By contrast, the improved insulin re- sponsiveness and sensitivity of glucose uptake were observed in L6-GLUT4myc myoblasts. Earlier studies indicated that forskolin might affect insulin-stimulated GLUT4 translocation. A 65% decrease of insulin-stimulated 2-deoxyglucose uptake in GLUT4myc cells was not due to an effect on GLUT4 mobi- lization to the plasma membrane, but instead on direct inhibition of GLUT4. Forskolin and dipyridamole are more potent inhibitors of GLUT4 than GLUT1. Alternatively, pentobarbital inhibits GLUT1 more than GLUT4. The use of these inhibitors confirmed that the overexpressed GLUT1 or GLUT4 are the major functional glucose transporters in unstimulated and insulin-stimulated L6 myoblasts. Therefore, L6-GLUT1myc and L6-GLUT4myc cells provide a platform to screen compounds that may have differ- ential effects on GLUT isoform activity or may influence GLUT isoform mobilization to the cell surface of muscle cells.

关键词： L6 muscle cells, glucose transporter (GLUT), insulin, 2-deoxyglucose uptake

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插入到细胞膜上的葡萄糖转运子4可不暴露其被光化学法标记的活性位点

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中国科学（C辑） 2006年第6期36卷 508-515页

作者：牛文彦 Manabu Ishiki philip j bilan 姚智天津医科大学免疫学教研室天津300070 Programme in Cell Biology The Hospital for Sick ChildrenTorontoCanada M5G 1X8

胰岛素刺激骨胳肌产生磷脂酰肌醇3,4,5三磷酸(PI(3,4,5)P3),它是促进葡萄糖转运子4(GLUT4)与细胞膜融合的必要条件.向肌肉细胞内导入PI(3,4,5)P3可以模拟胰岛素刺激GLUT4与细胞膜融合的作用,但不足以增加细胞摄取葡萄糖的量.本研究目的... 详细信息

胰岛素刺激骨胳肌产生磷脂酰肌醇3,4,5三磷酸(PI(3,4,5)P3),它是促进葡萄糖转运子4(GLUT4)与细胞膜融合的必要条件.向肌肉细胞内导入PI(3,4,5)P3可以模拟胰岛素刺激GLUT4与细胞膜融合的作用,但不足以增加细胞摄取葡萄糖的量.本研究目的是探讨PI(3,4,5)P3与胰岛素作用不同的机制.在骨骼肌细胞株(L6-GLUT4myc)中,应用免疫反应方法检测细胞膜片上与特异性抗体反应的GLUT4的胞浆区羧基末端表位和胞外区myc表位的可用性;使用不能渗透到细胞内的甘露糖-生物素衍生物Bio-LC-ATB-BMPA,结合亲和光化学标记法检测GLUT4胞外区的活性位点.相对于基础组,100nmol/L胰岛素和10μmol/LPI(3,4,5)P3分别使与myc结合的抗体量增加1.64倍和1.58倍.胰岛素还使细胞膜上GLUT4的光化学标记量和细胞膜片上与羧基末端表位结合的抗体量分别增加了2.47倍和2.04倍,而PI(3,4,5)P3则无此作用.在胰岛素作用下,细胞膜片上与羧基末端表位结合的抗体量大于与myc表位结合的抗体量(分别为2.04和1.64倍).结果表明:(i)尽管PI(3,4,5)P3能使GLUT4与细胞膜融合,但不能使GLUT4胞外区的活性位点暴露;(ii)GLUT4胞外区活性位点的可用性与胞浆区羧基末端的可用性相关;(iii)除了能刺激GLUT4与细胞膜融合,胰岛素还使封闭GLUT4羧基末端的蛋白脱离.推论胞浆内某种蛋白封闭羧基末端,同样阻止甘露糖-生物素衍生物对GLUT4活性位点的标记,并可能妨碍GLUT4转运葡萄糖.

关键词：葡萄糖转运子4(GLUT4) 胰岛素磷脂酰肌醇3,4,5三磷酸(PI(3,4,5)P3) 光化学标记

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葡萄糖转运的胰岛素敏感性以及forskolin,dipyridamole和pentobarbital对三种L6骨骼肌细胞葡萄糖转运的抑制作用

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中国科学（C辑） 2007年第4期37卷 402-409页

作者：牛文彦 philip j bilan Michiko Hayashi 笪宇蓉姚智天津医科大学免疫学教研室天津300070 Programmein CellBiology The Hospital for Sick ChildrenTorontoCanada M5G 1X8

用稳定过表达并带有myc表位的葡萄糖转运子1(glucose transporter1,GLUT1)或葡萄糖转运子4(glucose transporter4,GLUT4)的L6骨骼肌细胞株定征GLUT1和GLUT4对胰岛素的响应.所筛选的L6-GLUT1myc细胞克隆分化前后的葡萄糖摄取量均在线性范... 详细信息

用稳定过表达并带有myc表位的葡萄糖转运子1(glucose transporter1,GLUT1)或葡萄糖转运子4(glucose transporter4,GLUT4)的L6骨骼肌细胞株定征GLUT1和GLUT4对胰岛素的响应.所筛选的L6-GLUT1myc细胞克隆分化前后的葡萄糖摄取量均在线性范围.100nmol/L胰岛素使L6-GLUT1myc和L6-GLUT4myc肌原细胞膜上GLUT1或GLUT4的量分别达到基础组的(1.58±0.01)倍和(1.96±0.11)倍,2-脱氧葡萄糖摄取量分别达到了(1.53±0.09)倍和(1.86±0.17)倍,此作用可被渥曼青霉素(wortmannin)抑制.胰岛素刺激了此2种细胞中的Akt磷酸化.L6-GLUT1myc肌原细胞的葡萄糖摄取量对胰岛素浓度呈剂量依赖性,但与野生型细胞相比,其对胰岛素的敏感性和最大响应没有改变.但L6-GLUT4myc肌原细胞的葡萄糖摄取量对胰岛素的敏感性和最大响应均增加.以前的研究提示毛喉素(forskolin)可能影响胰岛素刺激的GLUT4转位.本研究表明,在L6-GLUT4myc细胞中,毛喉素使胰岛素刺激的葡萄糖摄取减少了65%,此作用是由它对GLUT4的直接抑制而不是由其对GLUT4转位的影响造成的.毛喉素和dipyridamole对GLUT4比对GLUT1有更强的抑制作用,而戊巴比妥(pentobarbital)对GLUT1的抑制作用强于GLUT4.应用这些抑制剂的结果表明、L6肌原细胞中基础状态下和胰岛素刺激状态下的葡萄糖主要由过表达的GLUT1或GLUT4转运.因此,L6-GLUT1myc和L6-GLUT4myc细胞株为筛查对肌肉细胞GLUT1或GLUT4的活性或转位有不同作用的化合物提供了一个平台.

关键词： L6骨骼肌细胞株葡萄糖转运子(glucose transporter,GLUT) 胰岛素 2-脱氧葡萄糖摄取

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Hydromechanical characterization of gas transport amidst uncertainty for underground nuclear explosion detection

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journal of Rock Mechanics and Geotechnical Engineering 2024年第6期16卷 2019-2032页

作者： Wenfeng Li Chelsea W.Neil j William Carey Meng Meng Luke P.Frash philip H.Stauffer Earth and Environmental Sciences Los Alamos National LaboratoryLos AlamosNM87544USA Department of Mineral Engineering New Mexico TechSocorroNM87801USA

Given the challenge of definitively discriminating between chemical and nuclear explosions using seismic methods alone,surface detection of signature noble gas radioisotopes is considered a positive identification of underground nuclear explosions(UNEs).However,the migration of signature radionuclide gases between the nuclear cavity and surface is not well understood because complex processes are involved,including the generation of complex fracture networks,reactivation of natural fractures and faults,and thermo-hydro-mechanical-chemical(THMC)coupling of radionuclide gas transport in the *** this study,we provide an experimental investigation of hydro-mechanical(HM)coupling among gas flow,stress states,rock deformation,and rock damage using a unique multi-physics triaxial direct shear rock testing *** testing system also features redundant gas pressure and flow rate measurements,well suited for parameter uncertainty *** porous tuff and tight granite samples that are relevant to historic UNE tests,we measured the Biot effective stress coefficient,rock matrix gas permeability,and fracture gas permeability at a range of pore pressure and stress *** Biot effective stress coefficient varies from 0.69 to 1 for the tuff,whose porosity averages 35.3%±0.7%,while this coefficient varies from 0.51 to 0.78 for the tight granite(porosity<1%,perhaps an underestimate).Matrix gas permeability is strongly correlated to effective stress for the granite,but not for the porous *** experiments reveal the following key engineering implications on transport of radionuclide gases post a UNE event:(1)The porous tuff shows apparent fracture dilation or compression upon stress changes,which does not necessarily change the gas permeability;(2)The granite fracture permeability shows strong stress sensitivity and is positively related to shear displacement;and(3)Hydromechanical coupling among stress states,rock damage,and gas flow appears to be

关键词： Underground nuclear explosion uncertainty quantification Radionuclide transport Biot effective stress coefficient Fracture permeability Matrix permeability

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Adaptive neural network control of a 2-DOF helicopter system considering input constraints and global prescribed performance

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Science China(Information Sciences) 2024年第7期67卷 224-239页

作者： Zhijia ZHAO jiale WU Zhijie LIU We HE C.L.philip CHEN School of Mechanical and Electrical Engineering Guangzhou University School of Intelligence Science and Technology and Key Laboratory of Intelligent Bionic Unmanned Systems of Ministry of Education University of Science and Technology Beijing School of Computer Science and Engineering South China University of Technology Pazhou Lab

In this study, an adaptive neural network(NN) control is proposed for nonlinear two-degree-offreedom(2-DOF) helicopter systems considering the input constraints and global prescribed ***, radial basis function NN(RBFNN) is employed to estimate the unknown dynamics of the helicopter system. Second, a smooth nonaffine function is exploited to approximate and address nonlinear constraint functions. Subsequently, a new prescribed function is proposed, and an original constrained error is transformed into an equivalent unconstrained error using the error transformation and barrier function transformation methods. The analysis of the established Lyapunov function proves that the controlled system is globally uniformly bounded. Finally, the simulation and experimental results on a constructed Quanser's test platform verify the rationality and feasibility of the proposed control.

关键词： adaptive NN control 2-DOF helicopter global prescribed performance input constraints

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Prehospital transdermal glyceryl trinitrate for ultra- acute ischaemic stroke: data from the RIGHT- 2 randomised sham- controlled ambulance trial

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Stroke & Vascular Neurology 2024年第1期9卷 38-49,I0035-I0046页

作者： jason philip Appleton Lisa j Woodhouse Craig S Anderson Sandeep Ankolekar Lesley Cala Mark Dixon Timothy j England Kailash Krishnan Grant Mair Keith W Muir john Potter Christopher I Price Marc Randall Thompson G Robinson Christine Roffe Else C Sandset jeffrey L Saver Angela Shone Aloysius Niroshan Siriwardena joanna M Wardlaw Nikola Sprigg philip M Bath Stroke University Hospitals Birmingham NHS Foundation TrustBirminghamUK Institute of Applied HealthResearch University of Birmingham College of Medical and Dental SciencesBirminghamUK Stroke Trials Unit Mental Health and Clinical NeurosciencesUniversity of NottinghamNottinghamUK Faculty of Medicine The George Institute for Global HealthSydneyNew South WalesAustralia The George Institute China at Peking University Health Science Center BeijingChina Neurology Royal Prince Alfred HospitalSydney Health PartnersSydneyNew South WalesAustralia Department of Neurology King's College Hospital NHS Foundation TrustLondonUK Faculty of Health and Medical Sciences University of Western AustraliaCrawleyWestern AustraliaAustralia East Midlands Ambulance Service NHS Trust NottinghamUK Stroke Queen's Medical CentreNottingham University Hospitals NHS TrustNottinghamUK Centre for Clinical Brain Sciences Dementia Research InstituteUniveristy of EdinburghEdinburghUK Institute of Neurology and Psychology University of GlasgowGlasgowUK Bob Champion Research and Education Building University of East AngliaNorwichUK Institute of Neuroscience Newcastle UniversityNewcastle upon TyneUK Department of Neurology Leeds Teaching Hospitals NHS TrustLeedsUK Department of Cardiovascular Sciences and NIHR Leicester Biomedical Research Centre University of LeicesterLeicesterUK Stroke Research in Stoke Institute for Science and Technology in MedicineKeele UniversityStoke-on TrentUK Department of Neurology Oslo University HospitalOsloNorway Research and Development Norwegian Air Ambulance FoundationOsloNorway Department of Neurology and Comprehensive Stroke Center David Geffen School of MedicineUCLALos AngelesCaliforniaUSA Research and Graduate Services University of NottinghamNottinghamUK Community and Health Research Unit University of LincolnLincolnUK

Background The effect of transdermal glyceryl trinitrate(GTN,a nitrovasodilator)on clinical outcome when administered before hospital admission in suspected stroke patients is ***,we assess the safety and efficacy of GTN in the prespecified subgroup of patients who had an ischaemic stroke within the Rapid Intervention with Glyceryl trinitrate in Hypertensive stroke Trial-2(RIGHT-2).Methods RIGHT-2 was an ambulance-based multicentre sham-controlled blinded-endpoint study with patients randomised within 4hours of *** primary outcome was a shift in scores on the modified Rankin scale(mRS)at day *** outcomes included death;a global analysis(Wei-Lachin test)containing Barthel Index,EuroQol-5D,mRS,telephone interview for cognitive status-modified and Zung depression scale;and neuroimaging-determined‘brain frailty’*** were reported as n(%),mean(SD),median[IQR],adjusted common OR(acOR),mean difference or Mann-Whitney difference(MWD)with 95%*** 597 of 1149(52%)patients had a final diagnosis of ischaemic stroke;age 75(12)years,premorbid mRS>2107(18%),Glasgow Coma Scale 14(2)and time from onset to randomisation 67[45,108]***‘brain frailty’was common:median score 2[2,3](range 0–3).At day 90,GTN did not influence the primary outcome(acOR for increased disability 1.15,95%CI 0.85 to 1.54),death or global analysis(MWD 0.00,95%CI-0.10 to 0.09).In subgroup analyses,there were non-significant interactions suggesting GTN may be associated with more death and dependency in participants randomised within 1hour of symptom onset and in those with more severe *** In patients who had an ischaemic stroke,ultra-acute administration of transdermal GTN in the ambulance did not improve clinical outcomes in a population with more clinical and radiological frailty than seen in previous in-hospital *** IS ALREADY KNOWN ON THIS TOPIC⇒Transdermal glyceryl trinitrate(GTN)was associat-ed with less death and dependency in those w

关键词： Glasgow nitrate random

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Distinct vibrational signatures and complex phase behavior in metallic oxygen

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Matter and Radiation at Extremes 2024年第2期9卷 4-11页

作者： philip Dalladay-Simpson Bartomeu Monserrat Li Zhang Federico Gorelli Center for High Pressure Science Technology Advanced Research 1690 Cailun RoadShanghai 201203China Department of Materials Science and Metallurgy University of Cambridge27 Charles Babbage RoadCambridge CB3 OFSUnited Kingdom Cavendish Laboratory University of CambridgeJ.J.Thomson AvenueCambridge CB3 OHEUnited Kingdom Istituto Nazionale di Ottica Consiglio Nazionale delle Ricerche(CNR-INO)50125 FlorenceItaly Shanghai Advanced Research in Physical Sciences(SHARPS) ShanghaiChina

Evidence for metallization in dense oxygen has been reported for over 30 years[Desgreniers et al.,***.94,1117(1990)]at a now routinely accessible 95 GPa[Shimizu et al.,Nature 393,767(1998)].However,despite the longevity of this result and the technological advances since,the nature of the metallic phase remains poorly constrained[Akahama et al.,***.74,4690(1995);Goncharov et al.,***.B 68,224108(2003);Ma,***.B 76,064101(2007);and Weck et al.,***.102,255503(2009)].In this work,through Raman spectroscopy,we report the distinct vibrational characteristics of metallicζ-O_(2) from 85 to 225 *** comparison with numerical simulations,weﬁnd reasonable agreement with the candidate structure up to about 150 *** higher pressures,the C2/mstructure is found to be unstable and incompatible with experimental *** candidate structures,and Ci,with C2/m C2/conly two molecules in the primitive unit cell,are found to be stable and more compatible with measurements above 175 GPa,indicative of the dissociation of(O_(2))4 ***,we report and discuss a strong hysteresis and metastability with the precursory phaseϵ-O_(2).Theseﬁndings will reinvigorate experimental and theoretical work into the dense oxygen system,which will have importance for oxygen-bearing chemistry,prevalent in the deep Earth,as well as fundamental physics.

关键词： metallic phase vibrational

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Lipid nanoparticles deliver mRNA to the blood-brain barrier

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Nano Research 2024年第10期17卷 9126-9134页

作者： Yanina Kuzminich Avraham Shakked Randi Calkins Sebastian Rudden Camille jones jessie Doan Bora jang Elisa Schrader Echeverri Ryan Zenhausern Liming Lian David Loughrey Hannah E.Peck Rachelle Wiese Dorothy Koveal philip j.Santangelo james E.Dahlman George C.Woodruff School of Mechanical Engineering Georgia Institute of TechnologyAtlantaGA 30332USA Parker H.Petit Institute for Bioengineering and Biosciences Georgia Institute of TechnologyAtlantaGA 30332USA Wallace H.Coulter Department of Biomedical Engineering Emory University School of Medicine and Georgia Institute of TechnologyAtlantaGA 30332USA

Lipid nanoparticles(LNPs)have delivered RNA to hepatocytes in patients after intravenous *** clinical data support efforts to design LNPs that transfect cells in the central nervous system(CNS).However,delivery to the CNS has been difficult,in large part because quantifying on-target delivery alongside common off-target cell types in adult mice remains *** we report methods to isolate different cell types from the CNS,and subsequently present mRNA delivery readouts using a liver-detargeted *** data suggest that LNPs without targeting ligands can transfect cerebral endothelial cells in mice after intravenous *** the difficulty of crossing the blood-brain barrier,they also underscore the value of quantifying delivery in the CNS with cell-type resolution instead of whole-tissue resolution.

关键词： lipid nanoparticle mRNA nanomedicine central nervous system blood-brain barrier

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Boosting Adaptive Weighted Broad Learning System for Multi-Label Learning

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IEEE/CAA journal of Automatica Sinica 2024年第11期11卷 2204-2219页

作者： Yuanxin Lin Zhiwen Yu Kaixiang Yang Ziwei Fan C.L.philip Chen the School of Computer Science and Engineering in South China University of Technology IEEE the Pengcheng Laboratory

Multi-label classification is a challenging problem that has attracted significant attention from researchers, particularly in the domain of image and text attribute annotation. However, multi-label datasets are prone to serious intra-class and inter-class imbalance problems, which can significantly degrade the classification performance. To address the above issues, we propose the multi-label weighted broad learning system(MLW-BLS) from the perspective of label imbalance weighting and label correlation mining. Further, we propose the multi-label adaptive weighted broad learning system(MLAW-BLS) to adaptively adjust the specific weights and values of labels of MLW-BLS and construct an efficient imbalanced classifier set. Extensive experiments are conducted on various datasets to evaluate the effectiveness of the proposed model, and the results demonstrate its superiority over other advanced approaches.

关键词： Learning systems Adaptation models Correlation Adaptive systems Annotations Boosting Classification algorithms Broad learning system label correlation mining label imbalance weighting multi-label imbalance CLASSIFICATION ENSEMBLE

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