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https://www.readbyqxmd.com/read/28202688/super-resolution-mapping-of-scaffold-nucleoporins-in-the-nuclear-pore-complex
#1
Jiong Ma, Joseph M Kelich, Samuel L Junod, Weidong Yang
The nuclear pore complex (NPC), composed of ∼30 different nucleoporins (Nups), is one of the largest supramolecular structures in eukaryotic cells. Its octagonal ring-scaffold perforates the nuclear envelope and features a unique molecular machinery that regulates nucleocytoplasmic transport. However, the precise copy number and the spatial location of each Nup in the native NPC remain obscure due to the inherent difficulty of counting and localizing proteins inside the sub-micrometer supramolecular complex...
February 15, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/28202367/exosomes-therapy-delivery-tools-and-biomarkers-of-diseases
#2
REVIEW
Lucio Barile, Giuseppe Vassalli
Virtually all cells in the organism secrete extracellular vesicles (EVs), a heterogeneous population of lipid bilayer membrane-enclosed vesicles that transport and deliver payloads of proteins and nucleic acids to recipient cells, thus playing central roles in cell-cell communications. Exosomes, nanosized EVs of endosomal origin, regulate many pathophysiological processes including immune responses and inflammation, tumour growth, and infection. Healthy subjects and patients with different diseases release exosomes with different RNA and protein contents into the circulation, which can be measured as biomarkers...
February 12, 2017: Pharmacology & Therapeutics
https://www.readbyqxmd.com/read/28193881/strong-signal-increase-in-sted-fluorescence-microscopy-by-imaging-regions-of-subdiffraction-extent
#3
Fabian Göttfert, Tino Pleiner, Jörn Heine, Volker Westphal, Dirk Görlich, Steffen J Sahl, Stefan W Hell
Photobleaching remains a limiting factor in superresolution fluorescence microscopy. This is particularly true for stimulated emission depletion (STED) and reversible saturable/switchable optical fluorescence transitions (RESOLFT) microscopy, where adjacent fluorescent molecules are distinguished by sequentially turning them off (or on) using a pattern of light formed as a doughnut or a standing wave. In sample regions where the pattern intensity reaches or exceeds a certain threshold, the molecules are essentially off (or on), whereas in areas where the intensity is lower, that is, around the intensity minima, the molecules remain in the initial state...
February 13, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28155617/von-willebrand-factor-and-cardiovascular-disease-from-a-biochemical-marker-to-an-attractive-therapeutic-target
#4
Felice Gragnano, Enrica Golia, Francesco Natale, Renatomaria Bianchi, Ivana Pariggiano, Mario Crisci, Vincenzo Diana, Fabio Fimiani, Giuseppe Limongelli, Mariagiovanna Russo, Plinio Cirillo, Paolo Calabrò
Despite recent advances, there is still an unmet need in antithrombotic therapy. New drugs have to overcome old targets, looking for new complementary strategies to counteract thrombus formation and propagation. Since its initial recognition in the 50's, von Willebrand Factor (VWF) has proved to be a contributor in clot formation. The contribution of VWF to platelet adhesion and aggregation is pivotal at high shear rates (i.e. microcirculation and critical artery stenosis), where globular-inactive-VWF elongates in a long chain-active conformation...
February 1, 2017: Current Vascular Pharmacology
https://www.readbyqxmd.com/read/28152380/limiting-the-protein-corona-a-successful-strategy-for-in%C3%A2-vivo-active-targeting-of-anti-her2-nanobody-functionalized-nanostars
#5
Antoine D'Hollander, Hilde Jans, Greetje Vande Velde, Charlotte Verstraete, Sam Massa, Nick Devoogdt, Tim Stakenborg, Serge Muyldermans, Liesbet Lagae, Uwe Himmelreich
Gold nanoparticles hold great promise as anti-cancer theranostic agents against cancer by actively targeting the tumor cells. As this potential has been supported numerously during in vitro experiments, the effective application is hampered by our limited understanding and control of the interactions within complex in vivo biological systems. When these nanoparticles are exposed to a biological environment, their surfaces become covered with proteins and biomolecules, referred to as the protein corona, reducing the active targeting capabilities...
January 6, 2017: Biomaterials
https://www.readbyqxmd.com/read/28140525/nanobodies-against-surface-biomarkers-enable-the-analysis-of-tumor-genetic-heterogeneity-in-uveal-melanoma-patient-derived-xenografts
#6
Ronan Crépin, David Gentien, Angeline Duché, Audrey Rapinat, Cecile Reyes, Fariba Némati, Gérald Massonnet, Didier Decaudin, Selma Djender, Sandrine Moutel, Klervi Desrumeaux, Nathalie Cassoux, Sophie Piperno-Neumann, Sebastian Amigorena, Franck Perez, Sergio Roman Roman, Ario de Marco
Monoclonal antibodies specific for biomarkers expressed on the surface of uveal melanoma (UM) cells would simplify the immune-capture and genomic characterization of heterogeneous tumor cells originated from patient derived xenografts (PDXs). Antibodies against four independent tumor antigens were isolated by panning a nanobody synthetic library. Such antibodies enabled flow-cytometry-based sorting of distinct cell sub-populations from UM PDXs and to analyze their genomic features. The complexity and specificity of the biochemical and genomic biomarker combinations mirrored the UM tumor polyclonality...
January 31, 2017: Pigment Cell & Melanoma Research
https://www.readbyqxmd.com/read/28114943/joining-the-in-vitro-immunization-of-alpaca-lymphocytes-and-phage-display-rapid-and-cost-effective-pipeline-for-sdab-synthesis
#7
Lubos Comor, Saskia Dolinska, Katarina Bhide, Lucia Pulzova, Irene Jiménez-Munguía, Elena Bencurova, Zuzana Flachbartova, Lenka Potocnakova, Evelina Kanova, Mangesh Bhide
BACKGROUND: Camelids possess unique functional heavy chain antibodies, which can be produced and modified in vitro as a single domain antibody (sdAb or nanobody) with full antigen binding ability. Production of sdAb in conventional manner requires active immunization of Camelidae animal, which is laborious, time consuming, costly and in many cases not feasible (e.g. in case of highly toxic or infectious antigens). RESULTS: In this study, we describe an alternative pipeline that includes in vitro stimulation of naïve alpaca B-lymphocytes by antigen of interest (in this case endothelial cell binding domain of OspA of Borrelia) in the presence of recombinant alpaca interleukins 2 and 4, construction of sdAb phage library, selection of antigen specific sdAb expressed on phages (biopanning) and confirmation of binding ability of sdAb to the antigen...
January 23, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28107910/nanobody-based-enzyme-immunoassay-for-ochratoxin-a-in-cereal-with-high-resistance-to-matrix-interference
#8
Xing Liu, Zongwen Tang, Zhenhua Duan, Zhenyun He, Mei Shu, Xianxian Wang, Shirley J Gee, Bruce D Hammock, Yang Xu
A sensitive indirect competitive nanobody-based enzyme linked immunosorbent assay (Nb-ELISA) for ochratoxin A (OTA) with high resistance to cereal matrix interference was developed. Nanobodies against OTA (Nb15, Nb28, Nb32, Nb36) were expressed in E. coli cells and their thermal stabilities were compared with that of an OTA-specific monoclonal antibody 6H8. All nanobodies could still retain their antigen-binding activity after exposure to temperature 95°C for 5min or to 90°C for 75min. Nb28 that exhibited the highest sensitivity in ELISA was selected for further research...
March 1, 2017: Talanta
https://www.readbyqxmd.com/read/28065273/screening-and-characterization-strategies-for-nanobodies-targeting-membrane-proteins
#9
S Veugelen, M Dewilde, B De Strooper, L Chávez-Gutiérrez
The study of membrane protein function and structure requires their successful detection, expression, solubilization, and/or reconstitution, which poses a challenging task and relies on the availability of suitable tools. Several research groups have successfully applied Nanobodies in the purification, as well as the functional and structural characterization of membrane proteins. Nanobodies are small, single-chain antibody fragments originating from camelids presenting on average a longer CDR3 which enables them to bind in cavities and clefts (such as active and allosteric sites)...
2017: Methods in Enzymology
https://www.readbyqxmd.com/read/28055308/binding-to-ocular-albumin-as-a-half-life-extension-principle-for-intravitreally-injected-drugs-evidence-from-mechanistic-rat-and-rabbit-studies
#10
Holger Fuchs, Frederik Igney
PURPOSE: The ocular half-life of intravitreally (IVT) injected drugs is of major relevance for the suitability of a drug intended for chronic intraocular treatment, as the half-life determines the dosing frequency. Thus, half-life extension principles are very attractive as they can reduce the IVT dosing frequency. In this study, we investigated the ocular pharmacokinetics (PK) of the IVT injected Nanobody(®) BI-X and whether the noncovalent binding of BI-X to vitreous albumin could increase its ocular half-life...
January 5, 2017: Journal of Ocular Pharmacology and Therapeutics
https://www.readbyqxmd.com/read/27982020/intrabody-targeting-vascular-endothelial-growth-factor-receptor-2-mediates-downregulation-of-surface-localization
#11
E Alirahimi, A Ashkiyan, F Kazemi-Lomedasht, K Azadmanesh, M Hosseininejad-Chafi, M Habibi-Anbouhi, R Moazami, M Behdani
Angiogenesis is among the most important mechanisms that helps cancer cells to survive, grow and undergo metastasis. Therefore, inhibiting angiogenesis will suppress tumor growth. Vascular endothelial growth factor (VEGF) and its receptor (VEGFR) are believed to be important players of angiogenesis. The goal of this study was to evaluate the success of a novel nanobody against VEGFR2 in tethering its target inside the endoplasmic reticulum and preventing its transport to the cell membrane. Nanobody sequence was cloned in a mammalian vector in fusion with green fluorescent protein and a KDEL retention signal...
December 16, 2016: Cancer Gene Therapy
https://www.readbyqxmd.com/read/27959623/nanobodies-to-study-g-protein-coupled-receptor-structure-and-function
#12
Aashish Manglik, Brian K Kobilka, Jan Steyaert
Ligand-induced activation of G protein-coupled receptors (GPCRs) is a key mechanism permitting communication between cells and organs. Enormous progress has recently elucidated the structural and dynamic features of GPCR transmembrane signaling. Nanobodies, the recombinant antigen-binding fragments of camelid heavy-chain-only antibodies, have emerged as important research tools to lock GPCRs in particular conformational states. Active-state stabilizing nanobodies have elucidated several agonist-bound structures of hormone-activated GPCRs and have provided insight into the dynamic character of receptors...
January 6, 2017: Annual Review of Pharmacology and Toxicology
https://www.readbyqxmd.com/read/27940234/construction-of-iron-polymer-graphene-nanocomposites-with-low-nonspecific-adsorption-and-strong-quenching-ability-for-competitive-immunofluorescent-detection-of-biomarkers-in-gm-crops
#13
Kaifei Yin, Anran Liu, Li Shangguan, Li Mi, Xu Liu, Yuanjian Liu, Yuewu Zhao, Ying Li, Wei Wei, Yuanjian Zhang, Songqin Liu
We developed a new immunofluorescent biosensor by utilizing a novel nanobody (Nb) and iron-polymer-graphene nanocomposites for sensitive detection of 5-enolpyruvylshikimate-3-phosphate synthase from Agrobacdterium tumefaciens strain CP4 (CP4-EPSPS), which considered as biomarkers of genetically modified (GM) crops. Specifically, we prepared iron doped polyacrylic hydrazide modified reduced graphene nanocomposites (Fe@RGO/PAH) by in-situ polymerization approach and subsequent a one-pot reaction with hydrazine...
April 15, 2017: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/27917276/preparation-and-characterization-of-a-novel-nanobody-against-t-cell-immunoglobulin-and-mucin-3-tim-3
#14
Vida Homayouni, Mazdak Ganjalikhani-Hakemi, Abbas Rezaei, Hossein Khanahmad, Mahdi Behdani, Fatemeh Kazemi Lomedasht
OBJECTIVES: As T-cell immunoglobulin and mucin domain 3 (TIM-3) is an immune regulatory molecule; its blocking or stimulating could alter the pattern of immune response towards a desired condition. Based on the unique features of nanobodies, we aimed to construct an anti-TIM-3 nanobody as an appropriate tool for manipulating immune responses for future therapeutic purposes. MATERIALS AND METHODS: We immunized a camel with TIM-3 antigen and then, synthesized a VHH phage: mid library from its B cell's transcriptome using nested PCR...
November 2016: Iranian Journal of Basic Medical Sciences
https://www.readbyqxmd.com/read/27895170/prevention-of-v%C3%AE-9v%C3%AE-2-t-cell-activation-by-a-v%C3%AE-9v%C3%AE-2-tcr-nanobody
#15
Renée C G de Bruin, Anita G M Stam, Anna Vangone, Paul M P van Bergen En Henegouwen, Henk M W Verheul, Zsolt Sebestyén, Jürgen Kuball, Alexandre M J J Bonvin, Tanja D de Gruijl, Hans J van der Vliet
Vγ9Vδ2 T cell activation plays an important role in antitumor and antimicrobial immune responses. However, there are conditions in which Vγ9Vδ2 T cell activation can be considered inappropriate for the host. Patients treated with aminobisphosphonates for hypercalcemia or metastatic bone disease often present with a debilitating acute phase response as a result of Vγ9Vδ2 T cell activation. To date, no agents are available that can clinically inhibit Vγ9Vδ2 T cell activation. In this study, we describe the identification of a single domain Ab fragment directed to the TCR of Vγ9Vδ2 T cells with neutralizing properties...
January 1, 2017: Journal of Immunology: Official Journal of the American Association of Immunologists
https://www.readbyqxmd.com/read/27889311/structural-evaluation-of-a-nanobody-targeting-complement-receptor-vsig4-and-its-cross-reactivity
#16
Yurong Wen, Zhenlin Ouyang, Steve Schoonooghe, Siyu Luo, Patrick De Baetselier, Wuyuan Lu, Serge Muyldermans, Geert Raes, Fang Zheng
Vsig4 is a recently identified immune regulatory protein related to the B7 family with dual functionality: a negative regulator of T cell activation and a receptor for the complement components C3b and C3c. Here we present a structural evaluation of a nanobody, Nb119, against the extracellular IgV domain protein of both mouse and human recombinant Vsig4, which have a high degree of sequence identity. Although mouse and human Vsig4 bind to Nb119 with a 250 times difference in dissociation constants, the interaction results in a highly identical assembly with a RMSD of 0...
November 18, 2016: Immunobiology
https://www.readbyqxmd.com/read/27881823/nanobodies-that-block-gating-of-the-p2x7-ion-channel-ameliorate-inflammation
#17
Welbeck Danquah, Catherine Meyer-Schwesinger, Björn Rissiek, Carolina Pinto, Arnau Serracant-Prat, Miriam Amadi, Domenica Iacenda, Jan-Hendrik Knop, Anna Hammel, Philine Bergmann, Nicole Schwarz, Joana Assunção, Wendy Rotthier, Friedrich Haag, Eva Tolosa, Peter Bannas, Eric Boué-Grabot, Tim Magnus, Toon Laeremans, Catelijne Stortelers, Friedrich Koch-Nolte
Ion channels are desirable therapeutic targets, yet ion channel-directed drugs with high selectivity and few side effects are still needed. Unlike small-molecule inhibitors, antibodies are highly selective for target antigens but mostly fail to antagonize ion channel functions. Nanobodies-small, single-domain antibody fragments-may overcome these problems. P2X7 is a ligand-gated ion channel that, upon sensing adenosine 5'-triphosphate released by damaged cells, initiates a proinflammatory signaling cascade, including release of cytokines, such as interleukin-1β (IL-1β)...
November 23, 2016: Science Translational Medicine
https://www.readbyqxmd.com/read/27852863/cryo-electron-microscopy-structures-of-expanded-poliovirus-with-vhhs-sample-the-conformational-repertoire-of-the-expanded-state
#18
Mike Strauss, Lise Schotte, Krishanthi S Karunatilaka, David J Filman, James M Hogle
: By using cryo-electron microscopy, expanded 80S-like poliovirus virions (poliovirions) were visualized in complexes with four 80S-specific camelid VHHs (Nanobodies). In all four complexes, the VHHs bind to a site on the top surface of the capsid protein VP3, which is hidden in the native virus. Interestingly, although the four VHHs bind to the same site, the structures of the expanded virus differ in detail in each complex, suggesting that each of the Nanobodies has sampled a range of low-energy structures available to the expanded virion...
February 1, 2017: Journal of Virology
https://www.readbyqxmd.com/read/27832973/neutralization-of-tnf%C3%AE-in-tumor-with-a-novel-nanobody-potentiates-paclitaxel-therapy-and-inhibits-metastasis-in-breast-cancer
#19
Xuemei Ji, Zhengxin Peng, Xiaorui Li, Zhonghui Yan, Yue Yang, Zheng Qiao, Yu Liu
Metastatic disease is the major cause of death from cancer, and immunotherapy and chemotherapy have had limited success in reversing its progression. Researchers have suggested that inflammatory factors in the tumor environment can promote cancer invasion and metastasis, stimulating cancer progression. Thus, novel strategies that target cytokines and modulate the tumor microenvironment may emerge as important approaches for treating metastatic breast cancer. Specific neutralization of pathogenic TNF signaling using a TNFα antibody has gained increasing attention...
February 1, 2017: Cancer Letters
https://www.readbyqxmd.com/read/27825023/two-dimensional-crystallization-of-the-mouse-serotonin-5-ht3a-receptor
#20
Jan Rheinberger, Ghérici Hassaine, Mohamed Chami, Henning Stahlberg, Horst Vogel, Xiaodan Li
The mouse serotonin 5-HT3A receptor is a homo-pentameric ligand-gated ion channel (pLGIC) mediating fast excitatory neurotransmission in the central nervous system. The molecular mechanism of ion permeation of 5-HT3A receptors triggered by the neurotransmitter serotonin is not yet fully understood. The recent X-ray structure of the mouse serotonin 5-HT3A receptor in complex with a stabilizing nanobody revealed for the first time the entire structure of a mammalian pLGIC in detergent. Structural information of the receptor in a lipid bilayer however is still limited primarily due to the lack of 2D crystals of the receptor in a lipid bilayer...
January 2017: Micron: the International Research and Review Journal for Microscopy
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