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Liqun He, Michael Vanlandewijck, Elisabeth Raschperger, Maarja Andaloussi Mäe, Bongnam Jung, Thibaud Lebouvier, Koji Ando, Jennifer Hofmann, Annika Keller, Christer Betsholtz
Pericytes, the mural cells of blood microvessels, regulate microvascular development and function and have been implicated in many brain diseases. However, due to a paucity of defining markers, pericyte identification and functional characterization remain ambiguous and data interpretation problematic. In mice carrying two transgenic reporters, Pdgfrb-eGFP and NG2-DsRed, we found that double-positive cells were vascular mural cells, while the single reporters marked additional, but non-overlapping, neuroglial cells...
October 11, 2016: Scientific Reports
Laura Mourino-Alvarez, Montserrat Baldan-Martin, Laura Gonzalez-Calero, Carlos Martinez-Laborde, Tamara Sastre-Oliva, Rafael Moreno-Luna, Luis F Lopez-Almodovar, Pedro L Sanchez, Francisco Fernandez-Aviles, Fernando Vivanco, Luis R Padial, Finn Akerstrom, Gloria Alvarez-Llamas, Fernando de la Cuesta, María G Barderas
BACKGROUND: The most common valve diseases are calcific aortic stenosis (AS) and aortic regurgitation (AR). The former is characterized by thickening of valve leaflets followed by progressive calcification, which produces progressive aortic valve (AV) narrowing, increased pressure afterload on the left ventricle (LV) and subsequent LV hypertrophy. On the other hand, AR is due to malcoaptation of the valve leaflets with resultant diastolic reflux of blood from aorta back to the LV producing volume and pressure overload and progressive LV dilatation...
September 26, 2016: International Journal of Cardiology
Paola Mora-Uribe, Camila Miranda-Cárdenas, Pablo Castro-Córdova, Fernando Gil, Iván Calderón, Juan A Fuentes, Paula I Rodas, Saeed Banawas, Mahfuzur R Sarker, Daniel Paredes-Sabja
Clostridium difficile is the causative agent of the most frequently reported nosocomial diarrhea worldwide. The high incidence of recurrent infection is the main clinical challenge of C. difficile infections (CDI). Formation of C. difficile spores of the epidemic strain R20291 has been shown to be essential for recurrent infection and transmission of the disease in a mouse model. However, the underlying mechanisms of how these spores persist in the colonic environment remains unclear. In this work, we characterized the adherence properties of epidemic R20291 spores to components of the intestinal mucosa, and we assessed the role of the exosporium integrity in the adherence properties by using cdeC mutant spores with a defective exosporium layer...
2016: Frontiers in Cellular and Infection Microbiology
Chew-Li Soh, Danwei Huangfu
The recent advent of engineered nucleases including the CRISPR/Cas9 system has greatly facilitated genome manipulation in human pluripotent stem cells (hPSCs). In addition to facilitating hPSC-based disease studies, the application of genome engineering in hPSCs has also opened up new avenues for cell replacement therapy. To improve consistency and reproducibility of hPSC-based studies, and to meet the safety and regulatory requirements for clinical translation, it is necessary to use a defined, xeno-free cell culture system...
2017: Methods in Molecular Biology
Maryam Riaz, Marie Versaevel, Danahe Mohammed, Karine Glinel, Sylvain Gabriele
Despite the importance of matrix rigidity on cell functions, many aspects of the mechanosensing process in highly migratory cells remain elusive. Here, we studied the migration of highly motile keratocytes on culture substrates with similar biochemical properties and rigidities spanning the range between soft tissues (~kPa) and stiff culture substrates (~GPa). We show that morphology, polarization and persistence of motile keratocytes are regulated by the matrix stiffness over seven orders of magnitude, without changing the cell spreading area...
September 28, 2016: Scientific Reports
Yan Ji, Zhen Weng, Philip Fish, Neha Goyal, Mao Luo, Samantha P Myears, Tammy L Strawn, Bysani Chandrasekar, Jianbo Wu, William P Fay
OBJECTIVE: Plasminogen activator inhibitor-1 (PAI-1), a serine protease inhibitor that promotes and inhibits cell migration, plays a complex and important role in adverse vascular remodeling. Little is known about the effects of pharmacological PAI-1 inhibitors, an emerging drug class, on migration of vascular smooth muscle cells (SMCs) and endothelial cells (ECs), crucial mediators of vascular remodeling. We investigated the effects of PAI-039 (tiplaxtinin), a specific PAI-1 inhibitor, on SMC and EC migration in vitro and vascular remodeling in vivo...
September 22, 2016: Arteriosclerosis, Thrombosis, and Vascular Biology
Masaharu Hazawa, Takeshi Yasuda, Ai Saotome-Nakamura, Kenichi Tomiyama, Chizuka Obara, Takaya Goto, Katsushi Tajima
Plasminogen activator inhibitor-1 (PAI-1) is induced by radiation resulting in endothelial cell impairment, potentially leading to multiple organ failure. Vitronectin (VN) is a 75-kDa glycoprotein (VN75) cleaved into two forms (VN75 or VN65/10) by furin, which is regulated by intracellular PAI-1. VN protects against radiation-induced endothelial cell death, but the mechanisms involved in VN processing and its interactions with intra- and extracellular PAI-1 remain unclear. We examined these processes in cells in vitro using recombinant proteins or overexpression of VN and PAI-1 genes, including furin-susceptible (T(381)) and furin-resistant VN (A(381))...
September 17, 2016: Vascular Pharmacology
Sheng Dai, Rong Li, Yan Long, Steve Titus, Jinghua Zhao, Ruili Huang, Menghang Xia, Wei Zheng
Human neuronal cells differentiated from induced pluripotent cells have emerged as a new model system for the study of disease pathophysiology and evaluation of drug efficacy. Differentiated neuronal cells are more similar in genetics and biological content to human brain cells than other animal disease models. However, culture of neuronal cells in assay plates requires a labor-intensive procedure of plate precoating, hampering its applications in high-throughput screening (HTS). We developed a simplified method with one-step seeding of neural stem cells in assay plates by supplementing the medium with a recombinant human vitronectin (VTN), thus avoiding plate precoating...
September 19, 2016: Journal of Biomolecular Screening
Gabriela Schneider, Ewa Bryndza, Agata Poniewierska-Baran, Karol Serwin, Malwina Suszynska, Zachariah P Sellers, Michael L Merchant, Alagammai Kaliappan, Janina Ratajczak, Magda Kucia, Nichola C Garbett, Mariusz Z Ratajczak
Diluted (1%) plasma induces migration of malignant cell lines much more strongly than potent pro-metastatic factors. To characterize the factor(s) present in diluted plasma responsible for this phenomenon we performed i) heat inactivation, ii) dialysis, iii) proteinase K treatment, and iv) molecular size filtration studies. We found that this remarkable pro-migratory activity of diluted normal plasma is associated with a ~50-100-kD protein that interacts with GαI protein-coupled receptors and activates p42/44 MAPK and AKT signaling in target cells...
September 13, 2016: Oncotarget
Li Xiao, Mengmeng Ding, Osama Saadoon, Eric Vess, Andrew Fernandez, Ping Zhao, Li Jin, Xudong Li
Tissue engineering provides a promising approach to treat degenerative disc disease, which usually requires a large quantity of seed cells. A simple and reliable in vitro culture system to expand seed cells in a timely fashion is necessary to implement the application clinically. Here, we sought to establish a cost-effective culture system for expanding human annulus fibrosus cells using extracellular matrix (ECM) proteins as culture substrates. Cells were cultured onto a plastic surface coated with various types of ECMs, including fibronectin, vitronectin, collagen type I, gelatin and cell-free matrix deposited by human nucleus pulposus cells...
September 13, 2016: Cell and Tissue Research
Jennifer A Haworth, Howard F Jenkinson, Helen J Petersen, Catherine R Back, Jane L Brittan, Steve W Kerrigan, Angela H Nobbs
A range of Streptococcus bacteria are able to interact with blood platelets to form a thrombus (clot). Streptococcus gordonii is ubiquitous within the human oral cavity and amongst the common pathogens isolated from subjects with infective endocarditis. Two cell surface proteins, Hsa and Platelet adherence protein A (PadA), in S. gordonii mediate adherence and activation of platelets. In this study, we demonstrate that PadA binds activated platelets and that an NGR (Asparagine-Glycine-Arginine) motif within a 657 amino acid residue N-terminal fragment of PadA is responsible for this, together with two other integrin-like recognition motifs RGT and AGD...
September 11, 2016: Cellular Microbiology
Anne Gründel, Enno Jacobs, Roger Dumke
Mycoplasma pneumoniae is a major cause of community-acquired respiratory infections worldwide. Due to the strongly reduced genome, the number of virulence factors expressed by this cell wall-less pathogen is limited. To further understand the processes during host colonization, we investigated the interactions of the previously confirmed surface-located glycolytic enzymes of M. pneumoniae (pyruvate dehydrogenase A-C [PdhA-C], glyceraldehyde-3-phosphate dehydrogenase [GapA], lactate dehydrogenase [Ldh], phosphoglycerate mutase [Pgm], pyruvate kinase [Pyk] and transketolase [Tkt]) to the human extracellular matrix (ECM) proteins fibrinogen (Fn), fibronectin (Fc), lactoferrin (Lf), laminin (Ln) and vitronectin (Vc), respectively...
September 3, 2016: International Journal of Medical Microbiology: IJMM
N V Andreeva, O G Leonova, V I Popenko, A V Belyavsky
Extracellular cell matrices deposited by cells stimulate cell proliferation. However, their generation is cumbersome and time consuming. We show here that controlled fixation of fibronectin layers after coating culture vessels significantly enhances expansion of murine and human mesenchymal stem cells (MSCs) and, to a lesser extent, primary fibroblasts. In contrast, fibronection fixation did not stimulate proliferation of established cancer cell lines. Fixed vitronectin or collagen IV layers also enhanced proliferation of murine MSCs...
December 1, 2016: Analytical Biochemistry
Yongshun Lin, Kaari L Linask, Barbara Mallon, Kory Johnson, Michael Klein, Jeanette Beers, Wen Xie, Yubin Du, Chengyu Liu, Yinzhi Lai, Jizhong Zou, Mark Haigney, Hushan Yang, Mahendra Rao, Guokai Chen
: : Cardiomyocytes can be differentiated from human pluripotent stem cells (hPSCs) in defined conditions, but efficient and consistent cardiomyocyte differentiation often requires expensive reagents such as B27 supplement or recombinant albumin. Using a chemically defined albumin-free (E8 basal) medium, we identified heparin as a novel factor that significantly promotes cardiomyocyte differentiation efficiency, and developed an efficient method to differentiate hPSCs into cardiomyocytes...
September 2, 2016: Stem Cells Translational Medicine
Gili Kaufman, Laiz Nunes, Alex Eftimiades, Wojtek Tutak
Tissue engineering-based therapies rely on the delivery of monolayered fibroblasts on two-dimensional polystyrene-coated and extracellular matrix (ECM) surfaces to regenerate connective tissues. However, this approach may fail to mimic their three-dimensional (3D) native architecture and function. We hypothesize that ECM fibrous proteins, which direct the migration of cells in vivo, may attach and guide polystyrene- and Matrigel™-ECM (M-ECM)-adherent fibroblasts to rearrangement into large multicellular macrostructures with the ability to proliferate...
August 30, 2016: Cells, Tissues, Organs
Jasper Foolen, Jau-Ye Shiu, Maria Mitsi, Yang Zhang, Christopher S Chen, Viola Vogel
Generating and maintaining gradients of cell density and extracellular matrix (ECM) components is a prerequisite for the development of functionality of healthy tissue. Therefore, gaining insights into the drivers of spatial organization of cells and the role of ECM during tissue morphogenesis is vital. In a 3D model system of tissue morphogenesis, a fibronectin-FRET sensor recently revealed the existence of two separate fibronectin populations with different conformations in microtissues, i.e. 'compact and adsorbed to collagen' versus 'extended and fibrillar' fibronectin that does not colocalize with the collagen scaffold...
2016: PloS One
Ivan Mikšík, Pavla Sedláková, Statis Pataridis, Federica Bortolotti, Rossella Gottardo
Proteins and their modifications of the natural mummy of Cangrande della Scala (Prince of Verona, Northern Italy, 1291-1329) were studied. The nano-LC-Q-TOF analysis of samples of rib bone and muscle from the mummy showed the presence of different proteins including type I, III IV, V and XI collagen, haemoglobin (subunits alpha and beta), ferritin, biglycan, vitronectin, prothrombin and osteocalcin. The structure of type I and type III collagen was deeply studied in order to evaluate the occurrence of modifications in comparison with type I and type III collagen coming from tissues of recently died people...
August 20, 2016: Protein Science: a Publication of the Protein Society
Pingbo Zhang, Rabia Karani, Randi L Turner, Craig Dufresne, Sara Ferri, Jennifer E Van Eyk, Richard D Semba
The optic nerve is a white matter tract that conveys visual information to the brain. The sclera comprises the white, protective outer layer of the eye. A characterization of the proteome of normal human retrobulbar optic nerve and sclera may facilitate studies of the eye. We conducted a proteomic analysis of optic nerve and sclera from five adults. Proteins were fractionated using SDS-PAGE. After in-gel digestion, peptides were analyzed using LC-MS/MS on an Orbitrap Elite mass spectrometer. We identified 2711 non-redundant proteins in retrobulbar optic nerve and 1945 non-redundant proteins in sclera...
October 2016: Proteomics
Carmine Selleri, Nunzia Montuori, Annamaria Salvati, Bianca Serio, Ada Pesapane, Patrizia Ricci, Anna Gorrasi, Anna Li Santi, Gunilla Hoyer-Hansen, Pia Ragno
Hematopoietic stem cells (HSCs) reside in bone marrow (BM) and can be induced to mobilize into the circulation for transplantation. Homing and lodgement into BM of transplanted HSCs are the first critical steps in their engraftment and involve multiple interactions between HSCs and the BM microenvironment. uPAR is a three domain receptor (DIDIIDIII) which binds urokinase, vitronectin, integrins. uPAR can be cleaved and shed from the cell surface generating full-length and cleaved soluble forms (suPAR and DIIDIII-suPAR)...
August 8, 2016: Oncotarget
Jonas Nascimento Conde, Emiliana Mandarano da Silva, Diego Allonso, Diego Rodrigues Coelho, Iamara da Silva Andrade, Luciano Neves de Medeiros, Joice Lima Menezes, Angela Silva Barbosa, Ronaldo Mohana-Borges
: Dengue virus (DENV) infects millions of people worldwide and is a major public health problem. DENV nonstructural protein 1 (NS1) is a conserved glycoprotein that associates with membranes and is also secreted into the plasma in DENV-infected patients. The present study describes a novel mechanism by which NS1 inhibits the terminal complement pathway. We first identified the terminal complement regulator vitronectin (VN) as a novel DENV2 NS1 binding partner by using a yeast two-hybrid system...
November 1, 2016: Journal of Virology
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