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RUFY4 deletion prevents pathological bone loss by blocking endo-lysosomal trafficking of osteoclasts

Bone Research 2024年第2期12卷 407-420页

作者： Minhee Kim Jin Hee Park Miyeon Go Nawon Lee Jeongin Seo Hana Lee Doyong Kim Hyunil Ha Taesoo Kim Myeong Seon Jeong Suree Kim Taesoo Kim Han Sung Kim Dongmin Kang Hyunbo Shim Soo Young Lee Department of Life Science Ewha Womans UniversitySeoul 03760South Korea The Research Center for Cellular Homeostasis Ewha Womans UniversitySeoul 03760South Korea Department of Biomedical Engineering Yonsei UniversityWonju 26493South Korea KM Convergence Research Division Korea Institute of Oriental MedicineDaejeon 34054South Korea Chuncheon Center Korea Basic Science InstituteChuncheon 24341South Korea Fluorescence core Imaging Center and bioimaging Data Curation Center Ewha Womans UniversitySeoul 03760South Korea Multitasking Macrophage Research Center Ewha Womans UniversitySeoul 03760South Korea

Mature osteoclasts degrade bone matrix by exocytosis of active proteases from secretory lysosomes through a ruffled ***,the molecular mechanisms underlying lysosomal trafficking and secretion in osteoclasts remain largely ***,we show with GeneChip analysis that RUN and FYVE domain-containing protein 4(RUFY4)is strongly upregulated during *** lacking Rufy4 exhibited a high trabecular bone mass phenotype with abnormalities in osteoclast function in ***,deleting Rufy4 did not affect osteoclast differentiation,but inhibited bone-resorbing activity due to disruption in the acidic maturation of secondary lysosomes,their trafficking to the membrane,and their secretion of cathepsin K into the extracellular ***,RUFY4 promotes late endosome-lysosome fusion by acting as an adaptor protein between Rab7 on late endosomes and LAMP2 on primary ***,Rufy4-deficient mice were highly protected from lipopolysaccharide-and ovariectomy-induced bone ***,RUFY4 plays as a new regulator in osteoclast activity by mediating endo-lysosomal trafficking and have a potential to be specific target for therapies against bone-loss diseases such as osteoporosis.

关键词： osteoclast inhibited traffic

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QT-GWAS:A novel method for unveiling biosynthetic loci affecting qualitative metabolic traits

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Molecular Plant 2023年第7期16卷 1212-1227页

作者： Marlies Brouckaert Meng Peng RenéHofer lias El Houari Chiarina Darrah Veronique Storme Yvan Saeys Ruben Vanholme Geert Goeminne Vitaliy I.Timokhin John Ralph Kris Morreel Wout Boerjan Ghent University Department of Plant Biotechnology and Bioinformatics9000 GhentBelgium vib Center for Plant Systems Biology 9052 GhentBelgium Ghent University Department of Applied MathematicsComputer Science and Statistics9000 GhentBelgium vib Center for Inflammation Research 9052 GhentBelgium vib Metabolomics core 9052 GhentBelgium Department of Biochemistry and US Department of Energy Great Lakes Bioenergy Research CenterWisconsin Energy InstituteUniversity of Wisconsin-MadisonMadisonWI53706USA Ghent University Department of Applied MathematicsComputer Science and StatisticsGhentBelgium vib Center for Inflammation Research GhentBelgium BioNTech SE MainzRhineland-PalatinateGermany Eunomia Research&Consulting BristolUK vib Agro-incubator NeveleBelgium Research Institute for Chromatography KortrijkBelgium

Although the plant kingdom provides an enormous diversity of metabolites with potentially beneficial applications for humankind,a large fraction of these metabolites and their biosynthetic pathways remain *** metabolite structures and their biosynthetic pathways is key to gaining biological understanding andto allow metabolic *** orderto retrieve novel biosynthetic genes involved in specialized metabolism,we developed a novel untargeted method designated as qualitative trait GWAs(QT-GWAS)that subjects qualitative metabolic traits to a genome-wide association study,while the conventional metabolite GWAS(mGWAS)mainly considers the quantitative variation of *** a proof of the validity of QT-GWAS,23 and 15of the retrieved associations identified in Arabidopsis thaliana by QTGWAS and mGWAS,respectively,were supported by previous ***,seven genemetabolite associations retrieved by QT-GWAS were confirmed in this study through reverse genetics combined with metabolomics and/or in vitro enzyme *** such,we established that CYTOCHROME P450706A5(CYP706A5)is involved in the biosynthesis of chroman derivatives,UDP-GLYCOSYLTRANSFERASE 76C3(UGT76C3)is able tohexosylate guanine in vitro and in planta,and SULFOTRANSFERASE 202B1(SULT202B1)catalyzes the sulfation of neolignans in ***,our study demonstrates that the untargeted QT-GWAS method can retrieve valid gene-metabolite associations at the level of enzyme-encoding genes,even new associations that cannot be found by the conventional mGwAs,providing a new approach for dissecting qualitative metabolic traits.

关键词： qualitative method proof

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

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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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A cellular hierarchy in melanoma uncouples growth and metastasis

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四川生理科学杂志 2022年第9期44卷 1633-1633页

作者： Panagiotis Karras Laboratory for Molecular Cancer Biology Center for Cancer Biology VIB Leuven Belgium Department of Oncology KU Leuven Leuven Belgium Laboratory of Tumor Microenvironment and Therapeutic Resistance Center for Cancer Biology VIB Leuven Belgium Department of Applied Mathematics and Theoretical Physics Centre for Mathematical Sciences University of Cambridge Cambridge UK The Wellcome Trust/CRUK Gurdon Institute University of Cambridge Cambridge UK Wellcome Trust-MRC Stem Cell Institute University of Cambridge Cambridge UK Department of General Medical Oncology UZ Leuven Leuven Belgium BGI-Shenzhen Shenzhen China vib bioimaging core VIB Center for Brain and Disease Research Leuven Belgium vib bioimaging core VIB Center for Inflammation Research Ghent Belgium Department of Biomedical Molecular Biology Ghent University Ghent Belgium FACS Expertise Center Center for Cancer Biology VIB Leuven Belgium Center for Brain & Disease Research VIB-KU Leuven Leuven Belgium Department of Human Genetics KU Leuven Leuven Belgium Data Mining and Modeling for Biomedicine Group VIB Center for Inflammation Research Ghent Belgium Department of Applied Mathematics Computer Science and Statistics Ghent University Ghent Belgium Laboratory for Translational Cell and Tissue Research Department of Imaging and Pathology KU Leuven Leuven Belgium Laboratory of Translational Genetics Center for Cancer Biology VIB Leuven Belgium Laboratory of Translational Genetics Center for Human Genetics KU Leuven Leuven Belgium Herbert Irving Comprehensive Center Columbia University Irving Medical Center New York USA Laboratory of Stem Cells and Cancer Université Libre de Bruxelles (ULB) Brussels Belgium

Although melanoma is notorious for its high degree of heterogeneity and plasticity1,2,the origin and magnitude of cell-state diversity remains poorly ***,it is unclear whether growth and metastatic dissemination are supported by overlapping or distinct melanoma ***,by combining mouse genetics,single-cell and spatial transcriptomics,lineage tracing and quantitative modelling,we provide evidence of a hierarchical model of tumour growth that mirrors the cellular and molecular logic underlying the cell-fate specification and differentiation of the embryonic neural *** show that tumorigenic competence is associated with a spatially localized perivascular niche,a phenotype acquired through an intercellular communication pathway established by endothelial *** with a model in which only a fraction of cells are fated to fuel growth,temporal single-cell tracing of a population of melanoma cells with a mesenchymal-like state revealed that these cells do not contribute to primary tumour growth but,instead,constitute a pool of metastatic initiating cells that switch cell identity while disseminating to secondary *** data provide a spatially and temporally resolved map of the diversity and trajectories of melanoma cell states and suggest that the ability to support growth and metastasis are limited to distinct pools of *** observation that these phenotypic competencies can be dynamically acquired after exposure to specific niche signals warrant the development of therapeutic strategies that interfere with the cancer cell reprogramming activity of such microenvironmental cues.

关键词： melanoma metastasis organs

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Developments in cell culture systems for human pluripotent stem cells

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World Journal of Stem Cells 2019年第11期11卷 968-981页

作者： Weiwei Liu Chunhao Deng Carlos Godoy-Parejo Yumeng Zhang Guokai Chen Centre of Reproduction Development and AgingFaculty of Health SciencesUniversity of MacaoMacao 999078China Institute of Translational Medicine Faculty of Health SciencesUniversity of MacaoMacao 999078China bioimaging and Stem Cell core Facility Faculty of Health SciencesUniversity of MacaoMacao 999078China

Human pluripotent stem cells(hPSCs)are important resources for cell-based therapies and pharmaceutical *** order to realize the potential of hPSCs,it is critical to develop suitable technologies required for specific *** hPSC technologies depend on cell culture,and are critically influenced by culture medium composition,extracellular matrices,handling methods,and culture *** review summarizes the major technological advances in hPSC culture,and highlights the opportunities and challenges in future therapeutic applications.

关键词： Human pluripotent stem cells Human embryonic stem cells Cell culture Culture medium Stem cell niche Signal transduction Embryoid bodies

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Implementation of FLIM and SIFT for improved intraoperative delineation of glioblastoma margin

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Chinese Optics Letters 2017年第9期15卷 20-24页

作者：林丹樱罗腾刘丽炜陆原刘少雄袁振屈军乐 Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Cuangdong Province College of Optoelectronic EngineeringShenzhen UniversityShenzhen 518060China Department of Dermatology The Sixth People's Hospital of ShenzhenShenzhen 518052China Department of Pathology The Sixth People's Hospital of ShenzhenShenzhen 518052China bioimaging core Faculty of Health SciencesUniversity of MacaoTaipaMaeau SAR China

The aim of this study is to develop a novel technique for improving the intraoperative margin assessment of glioblastoma by examining the total extrinsic extracellular matrix（ECM） with eosin staining using fluorescence lifetime imaging microscopy（FLIM） and scale-invariant feature transform（SIFT） descriptor analysis. Pseudocolor FLIM images obviously exhibit ECM distributions, changes in sequential sections, and different regions of interest. Meanwhile, SIFT descriptors are first utilized for the discrimination of glioblastoma margins by matching similar ECM regions and extracting keypoint orientations from FLIM images obtained from a series of continuous slices. The findings indicate that FLIM imaging with SIFT analysis of the total ECM is a promising method for improving intraoperative diagnosis of frozen and surgically excised brain specimen sections.

关键词： ECM Implementation of FLIM and SIFT for improved intraoperative delineation of glioblastoma margin

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The Anaphase-Promoting Complex/Cyclosome in Control of Plant Development

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Molecular Plant 2012年第6期5卷 1182-1194页

作者： Jefri Heyman Lieven De Veylder Department of Plant Systems Biology VIB and Department of Plant Biotechnology and Bioinformatics Ghent University Technologiepark 927 B-9052Gent Belgium

Temporal controlled degradation of key cell division proteins ensures a correct onset of the different cell cycle phases and exit from the cell division program. In light of the cell cycle, the Anaphase-Promoting Complex/Cyclosome （APC/C） is an important conserved multi-subunit ubiquitin ligase, marking targets for degradation by the 26S protea- some. However, whereas the APC/C has been studied extensively in yeast and mammals, only in the last decade has the plant APC/C started to unveil its secrets. Research results have shown the importance of the APC/C core complex and its activators during gametogenesis, growth, hormone signaling, symbiotic interactions, and endoreduplication onset. In addition, recently, the first plant APC/C inhibitors have been reported, allowing a fine-tuning of APC/C activity during the cell cycle. Together with the identification of the first APC/C targets, a picture emerges of APC/C activity being essential for many different developmental processes.

关键词： cell cycle CDC20 CCS52 endoreduplication gametogenesis ubiquitination.

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A PIID-DTBT based semi-conducting polymer dots with broad and strong optical absorption in the visible-light region： Highly effective contrast agents for multiscale and multi-spectral photoacoustic imaging

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Nano Research 2017年第1期10卷 64-76页

作者： Jian Zhang Haobin Chen Ting Zhou Limei Wang Duyang Gao Xuanjun Zhang Yubin Liu Changfeng Wu Zhen Yuan bioimaging core Faculty of Health Sciences University of Macao Taipa Macao SAR 999078 China State Key Laboratory on Integrated Optoelectronics College of Electronic Science and Engineering Jilin University Jilin 130012 China College of Optoelectronic Engineering Shenzhen University Shenzhen 518060 China College of Veterinary Medicine South China Agricultural University Guangzhou 510642 China Center for Drug Non-clinical Evaluation and Research Guangdong Biological Resources Institute Guangdong Academy of Sciences Guangzhou 510900 China

As a hybrid imaging technique, photoacoustic imaging （PAI） can provide multiscale morphological information of tissues, and the use of multi-spectral PAI （MSPAI） can recover the spatial distribution of chromophores of interest, such as hemoglobin within tissues. Herein, we developed a contrast agent that can very effectively combine multiscale PAI with MSPAI for a more comprehensive characterization of complex biological tissues. Specifically, we developed novel PIID-DTBT based semi-conducting polymer dots （Pdots） that show broad and strong optical absorption in the visible-light region （500-700 nm）. The performances of gold nanoparticles （GNPs） and gold nanorods （GNRs）, which have been verified as excellent photoacoustic contrast agents, were compared with that of the Pdots based on the multiscale PAI system. Both ex vivo and in vivo experiments demonstrated that the Pdots have better photoacoustic conversion efficiency at 532 nm than GNPs and showed similar photoacoustic performance with GNRs at 700 nm at the same mass concentration. Photostability and toxicity tests demonstrated that the Pdots are photostable and biocompatible. More importantly, an in vivo MSPAI experiment indicated that the Pdots have better photoacoustic performance than the blood and therefore the signals can be accurately extracted from the background of vascular-rich tissues. Our work demonstrates the great potential of Pdots as highly effective contrast agents for the precise localization of lesions relative to the blood vessels based on multiscale PAI and MSPAI.

关键词： nanoparticles polymer dots contrast agents photoacoustic imaging multiscale imaging multi-spectral imaging

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Micro-and nano-fiber probes for optical sensing,imaging,and stimulation in biomedical applications

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Photonics Research 2020年第11期8卷 1703-1724页

作者： Xia Yu Shuyan Zhang Malini Olivo Nanxi Li School of Instrumentation and Optoelectronic Engineering Beihang UniversityBeijing 100083China Singapore bioimaging Consortium Agency for ScienceTechnologyand Research(A*STAR)Singapore 138667Singapore Institute of Microelectronics Agency for ScienceTechnologyand Research(A*STAR)Singapore 138634Singapore

The flexibile nature of optical fiber enables it to offer remote-access capabilities,which could be used in many biomedical *** review focuses on different micro-and nano-structured fiber probes for applications in biosensing,imaging,and *** modifications to fiber could extend design freedom from waveguide optimization to functional material *** probes with optimized waveguide structures or integrated functional materials could achieve enhanced optical mode interaction with biosamples,and hence obtain ultrasensitive biosensors with a remarkably low limit of ***,bioimaging with a high spatial resolution can be obtained by engineering dispersion and nonlinearity of light propagation in the fiber core or designing a metal-coated tapered fiber tip with a sub-wavelength *** metasurfaces can be assembled on a fiber tip to achieve a large depth of focus and remove *** is also a compact solution to realize the precise delivery of light for in vivo applications,such as deep brain *** optical beam size,shape,and direction could be steered by the probe ***-and nano-technologies integrated with fiber contribute to various approaches to further improve detection limit,sensitivity,optical resolution,imaging depth,and stimulation precision.

关键词： fiber waveguide stimulation

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The effect of lithium on resting-state brain networks in patients with bipolar depression

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Journal of Translational Neuroscience 2016年第1期1卷 43-51页

作者： Chunhong Liu Xin Ma Yuan Zhen Yu Zhang Lirong Tang Feng Li Changle Tie Chuanyue Wang Beijing Key Laboratory of Mental Disorders Department of Psychiatry Beijing Anding Hospital Capital Medical University Beijing 100088 China Center of Brain Disorders Beijing Institute for Brain Disorders Beijing 100069 China bioimaging core Faculty of Health SciencesUniversity of Macao Macao SAR China

Objective: Although lithium has been a commonly prescribed neurotrophic/neuroprotective mood-stabilizing agents, its effect on spontaneous brain activity in patients with bipolar depression remains unclear. The aim of this study is to reveal the basic mechanism underlying the pathological influences of lithium on resting-state brain function of bipolar depression patients. Methods: 97 subjects including 9 bipolar depression patients with lithium treatment, 19 bipolar depression patients without lithium treatment and 69 healthy controls, were recruited to participate in this study. Amplitude of low-frequency fluctuation (ALFF) and fractional amplitude of low-frequency fluctuation (fALFF) were used to capture the changes of spontaneous brain activity among different groups. In addition, further analysis in terms of Hamilton Depression Rating Scale, the number of depressive episodes, and illness duration in pooled bipolar depression patients were conducted, which combined FLEF and fALEF to identify the basic neural features of bipolar depression patients. Results: It was observed from the imaging results that both the bipolar depression patients receiving lithium treatment and healthy control subjects showed significantly decreased ALFF/fALFF values in the right anterior cingulate cortex and right middle frontal gyrus compared to that from the bipolar depression patients without lithium treatmetn. The ALFF values of the right middle temporal gyrus was also found to be negative related to the number of depressive episode and the total episodes. Conclusions: Our findings suggested that the bipolar depression subjects were identified to have abnormal ALFF/ fALFF in the cortico-limbic systems, including regions like right anterior cingulate cortex, bilateral middle frontal gyrus, right orbital frontal gyrus, and right middle temporal gyrus. In addition, it was also revealed that the decreased ALFF/fALFF in the right anterior cingulate cortex and right middle frontal gyrus

关键词： bipolar depression lithium functional magnetic resonance imaging (fMRI) amplitude of low-frequency fluctuation (ALFF) fractional amplitude of low-frequency fluctuation (fALFF) bipolar depression lithium functional magnetic resonance imaging (fMRI) amplitude of low-frequency fluctuation (ALFF) fractional amplitude of low-frequency fluctuation (fALFF)

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