DOCSLIB.ORG

Neurofilament Medium Polypeptide (NFM) Protein Concentration Is

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Neurofilament Medium Polypeptide (NFM) Protein Concentration Is Neurofilament medium polypeptide (NFM) protein concentration is increased in CSF and serum samples from patients with brain injury MARTÍNEZ-MORILLO, Eduardo, CHILDS, Charmaine <http://orcid.org/0000- 0002-1558-5633>, GARCÍA, Belén Prieto, ÁLVAREZ MENÉNDEZ, Francisco V., ROMASCHIN, Alexander D., CERVELLIN, Gianfranco, LIPPI, Giuseppe and DIAMANDIS, Eleftherios P. Available from Sheffield Hallam University Research Archive (SHURA) at: http://shura.shu.ac.uk/9484/ This document is the author deposited version. You are advised to consult the publisher's version if you wish to cite from it. Published version MARTÍNEZ-MORILLO, Eduardo, CHILDS, Charmaine, GARCÍA, Belén Prieto, ÁLVAREZ MENÉNDEZ, Francisco V., ROMASCHIN, Alexander D., CERVELLIN, Gianfranco, LIPPI, Giuseppe and DIAMANDIS, Eleftherios P. (2015). Neurofilament medium polypeptide (NFM) protein concentration is increased in CSF and serum samples from patients with brain injury. Clinical Chemistry and Laboratory Medicine, 53 (10), 1575-1584. Copyright and re-use policy See http://shura.shu.ac.uk/information.html Sheffield Hallam University Research Archive http://shura.shu.ac.uk Clin Chem Lab Med 2015; aop Eduardo Mart í nez-Morillo , Charmaine Childs , Bel é n Prieto Garc í a , Francisco V. Á lvarez Men é ndez , Alexander D. Romaschin , Gianfranco Cervellin , Giuseppe Lippi and Eleftherios P. Diamandis * Neurofilament medium polypeptide (NFM) protein concentration is increased in CSF and serum samples from patients with brain injury DOI 10.1515/cclm-2014-0908 Methods: An ELISA from Elabscience was used. The selec- Received September 14 , 2014 ; accepted January 6 , 2015 tivity was evaluated using size-exclusion chromatography and mass spectrometry. Intra- and inter-batch coefficients of variation (CV) were also studied. Fifty-one CSF samples Abstract from 36 age-matched patients with hemorrhagic stroke (HS) (n = 30), ischemic stroke (IS) (n = 11) and healthy indi- Background: Brain injury is a medical emergency that viduals (n = 10) were assayed. In addition, serum samples needs to be diagnosed and treated promptly. Several pro- from healthy volunteers (n = 47), 68 serum samples from teins have been studied as biomarkers of this medical seven patients with HS, 106 serum samples from 12 condition. The aims of this study were to: 1) evaluate the patients with traumatic brain injury (TBI) and 68 serum selectivity and precision of a commercial ELISA kit for samples from 68 patients with mild traumatic brain injury neurofilament medium polypeptide (NFM) protein; and (mTBI) were also analyzed. 2) evaluate the concentration in cerebrospinal fluid (CSF) Results: NFM was identified in the chromatographic and serum of healthy individuals and patients with brain fraction with highest immunoreactivity. The intra- and damage. inter-batch CVs were ≤ 10% and ≤ 13%, respectively. The CSF-NFM concentration in HS was significantly higher (p < 0.0001) than in IS and controls. Serum NFM con- *Corresponding author: Eleftherios P. Diamandis, MD, PhD, FRCPC, centration ranged from 0.26 to 8.57 ng/mL in healthy FRSC, Pathology and Laboratory Medicine Department, Mount Sinai = Hospital, Suite 6-201, Box 32, 60 Murray Street, Toronto, ON M5T individuals (median 2.29), from 0.97 to 42.4 ng/mL in 3L9, Canada, Phone: + 1 416 586 8443, Fax: + 1 416 619 5521, HS (median = 10.8) and from 3.48 to 45.4 ng/mL in TBI E-mail: [email protected] ; Department of Clinical (median = 14.7). Finally, 44% of patients with mTBI had Biochemistry, University Health Network, Toronto, ON, Canada ; increased NFM concentration, with significantly higher and Department of Pathology and Laboratory Medicine, Mount Sinai levels (p = 0.01) in patients with polytrauma. Hospital, Toronto, ON, Canada Eduardo Mart í nez-Morillo: Lunenfeld-Tanenbaum Research Conclusions: To our knowledge this is the first study Institute, Joseph and Wolf Lebovic Health Complex, Mount Sinai describing increased NFM levels in CSF and serum from Hospital, Toronto, ON, Canada ; and Department of Clinical patients with brain damage. Biochemistry, Laboratory of Medicine, Hospital Universitario Central de Asturias, Oviedo, Spain Keywords: biomarker; brain injury; cerebrospinal fluid; Charmaine Childs: Centre for Health and Social Care Research, neurofilament; serum; stroke. Faculty of Health and Wellbeing, Sheffield Hallam University, Sheffield, UK Bel é n Prieto Garc í a and Francisco V. Á lvarez Men é ndez: Department of Clinical Biochemistry, Laboratory of Medicine, Hospital Universitario Central de Asturias, Oviedo, Spain Introduction Alexander D. Romaschin: Li Ka Shing Knowledge Institute, St. Michael ’ s Hospital, Department of Laboratory Medicine and The term “ brain injury ” refers to a potentially lethal Pathobiology, University of Toronto, Toronto, ON, Canada medical condition that includes two major types of brain Gianfranco Cervellin: Emergency Department, Academic Hospital of damage: 1) spontaneous, non-traumatic brain injury; and Parma, Parma, Italy Giuseppe Lippi: Laboratory of Clinical Chemistry and Hematology, 2) traumatic brain injury (TBI). The most common cause of Academic Hospital of Parma, Parma, Italy. http://orcid.org/0000- brain injury is by accidental injury to the head. Approxi- 0001-9523-9054 mately 1.7 million Americans suffer a TBI each year, and Brought to you by | Complejo Asistencial de Salamanca Authenticated Download Date | 8/7/15 10:00 AM 2 Mart í nez-Morillo et al.: NFM concentration is increased in brain injury 75% – 90% of those are classified as mild traumatic brain Materials and methods injury (mTBI) [1] . However, stroke is the most common form of non-traumatic brain injury, with 795,000 people ELISA kit for NFM protein in the US suffering a new or recurrent stroke each year. Ischemic stroke (IS) represents 87% of all stroke cases A commercial ELISA kit [Human Neurofi lament 3 (NEF3); Catalog No: whereas hemorrhagic stroke (HS) accounts for 13% of E-EL-H0740; from Elabscience Biotechnology Co., Ltd] was used to cases [2] . measure NFM protein in various biological fl uids. The ELISA kit uses capture and secondary monoclonal antibodies from rat and whole pro- A brain injury is usually a medical emergency that tein (natural, purifi ed protein) as standard material, according to the needs to be diagnosed and treated promptly, in order to information provided by the manufacturer. The reported limit of detec- improve the likelihood of patient survival and reduce the tion is 9.4 pg/mL; with an analytical measurement range from 15.6 to consequences of brain damage. For this purpose, several 1000 pg/mL. The intra-batch coeffi cient of variation (CV) is below 10%. proteins such as S100 calcium-binding protein B (S100B), The assay was performed according to the manufacturer ’ s instructions. neuron-specific enolase (NSE) and glial fibrillary acidic protein (GFAP) have been proposed as diagnostic and Size exclusion high-performance liquid chromatography prognostic biomarkers of TBI and stroke [3 – 6] . However, (SE-HPLC) given the complexity and heterogeneity of brain injury eti- ology, none of these proteins seem to be useful in these The SE-HPLC was performed using 0.1 M NaH2 PO4 /Na2 HPO4 , 0.15 M patients as single biomarkers. Therefore, it has been sug- NaCl (pH 7.0) buff er at a fl ow rate of 0.5 mL/min for 60 min with a gel gested that integrated panels of biomarkers with specific fi ltration column TSKgel G3000SW, 7.5 mm (ID) x 60 cm (L), 10 μ m and complementary characteristics may assist in the diag- (Tosoh Bioscience, King of Prussia, PA, USA). Liquid chromatogra- phy was performed in a HPLC 1100 series (Agilent Technologies) with nosis, risk assessment, treatment selection and prediction diode array detector (DAD), set at 254 nm (UV lamp). The system was of clinical outcomes of patients with brain damage. controlled with ChemStation soft ware (Agilent Technologies). One In our previous study [7] , we identified three novel hundred microliters of each sample (100 μ L loop) were loaded into biomarkers of brain injury: neurofilament medium pol- the column. Proteins contained in four diff erent samples (NFM stand- ypeptide (NFM), α -internexin ( α -Inx) and β -synuclein ard from ELISA kit, brain tissue extract from previous study [7] , CSF sample from patient with HS and no blood contamination and serum ( β -Syn) in the cerebrospinal fluid (CSF) of patients with sample from healthy individual) were subjected to chromatographic HS, by using a mass spectrometry-based assay. The aim separation. Fractions (0.5 mL) were collected every minute, from 18 of the current work was to perform a verification study of to 58 min, aft er the isocratic fl ow was started. The same volume of a one of these biomarkers using a more sensitive analytical Gel Filtration Standard (Catalog 151-1901, Bio-Rad Laboratories) was assay. The protein selected was NFM for three reasons: loaded before samples. 1) given its novelty (to our knowledge this is the first study evaluating NFM as biomarker of brain injury); 2) Mass spectrometry identifi cation the other two members of its family, neurofilament light and heavy polypeptides (NFL and NFH), have also been Three SE-HPLC fractions from brain tissue extract were analyzed in studied and identified as potential biomarkers of HS [8 – an LTQ Orbitrap XL (Thermo Scientifi c). Samples volume was reduced 11] ; and 3) there was a commercial ELISA kit available, from 400 to 50 μ L using a SpeedVac concentrator (Savant SC250EXP, with enough sensitivity (low pg/mL level) to detect this Thermo Scientifi c) and then processed as described previously [7] . protein in CSF and blood samples of patients with brain Briefl y, proteins were denatured, reduced, alkylated and trypsin digested. Peptides were analyzed by liquid chromatography coupled damage. to tandem mass spectrometry (LC-MS/MS), and the resulting spectra Before we assayed NFM protein in biological samples were searched separately against the Human UniprotKB-SwissProt from patients with brain injury, the commercial ELISA kit Database 2013_10 (containing 88266 forward protein sequences) by was validated, since we and others have drawn attention Mascot soft ware (version 2.2, Matrix Science).
Load more

Recommended publications
  • Defining Functional Interactions During Biogenesis of Epithelial Junctions
    ARTICLE Received 11 Dec 2015 | Accepted 13 Oct 2016 | Published 6 Dec 2016 | Updated 5 Jan 2017 DOI: 10.1038/ncomms13542 OPEN Defining functional interactions during biogenesis of epithelial junctions J.C. Erasmus1,*, S. Bruche1,*,w, L. Pizarro1,2,*, N. Maimari1,3,*, T. Poggioli1,w, C. Tomlinson4,J.Lees5, I. Zalivina1,w, A. Wheeler1,w, A. Alberts6, A. Russo2 & V.M.M. Braga1 In spite of extensive recent progress, a comprehensive understanding of how actin cytoskeleton remodelling supports stable junctions remains to be established. Here we design a platform that integrates actin functions with optimized phenotypic clustering and identify new cytoskeletal proteins, their functional hierarchy and pathways that modulate E-cadherin adhesion. Depletion of EEF1A, an actin bundling protein, increases E-cadherin levels at junctions without a corresponding reinforcement of cell–cell contacts. This unexpected result reflects a more dynamic and mobile junctional actin in EEF1A-depleted cells. A partner for EEF1A in cadherin contact maintenance is the formin DIAPH2, which interacts with EEF1A. In contrast, depletion of either the endocytic regulator TRIP10 or the Rho GTPase activator VAV2 reduces E-cadherin levels at junctions. TRIP10 binds to and requires VAV2 function for its junctional localization. Overall, we present new conceptual insights on junction stabilization, which integrate known and novel pathways with impact for epithelial morphogenesis, homeostasis and diseases. 1 National Heart and Lung Institute, Faculty of Medicine, Imperial College London, London SW7 2AZ, UK. 2 Computing Department, Imperial College London, London SW7 2AZ, UK. 3 Bioengineering Department, Faculty of Engineering, Imperial College London, London SW7 2AZ, UK. 4 Department of Surgery & Cancer, Faculty of Medicine, Imperial College London, London SW7 2AZ, UK.
    [Show full text]
  • Absence of NEFL in Patient-Specific Neurons in Early-Onset Charcot-Marie-Tooth Neuropathy Markus T
    ARTICLE OPEN ACCESS Absence of NEFL in patient-specific neurons in early-onset Charcot-Marie-Tooth neuropathy Markus T. Sainio, MSc, Emil Ylikallio, MD, PhD, Laura M¨aenp¨a¨a, MSc, Jenni Lahtela, PhD, Pirkko Mattila, PhD, Correspondence Mari Auranen, MD, PhD, Johanna Palmio, MD, PhD, and Henna Tyynismaa, PhD Dr. Tyynismaa [email protected] Neurol Genet 2018;4:e244. doi:10.1212/NXG.0000000000000244 Abstract Objective We used patient-specific neuronal cultures to characterize the molecular genetic mechanism of recessive nonsense mutations in neurofilament light (NEFL) underlying early-onset Charcot- Marie-Tooth (CMT) disease. Methods Motor neurons were differentiated from induced pluripotent stem cells of a patient with early- onset CMT carrying a novel homozygous nonsense mutation in NEFL. Quantitative PCR, protein analytics, immunocytochemistry, electron microscopy, and single-cell transcriptomics were used to investigate patient and control neurons. Results We show that the recessive nonsense mutation causes a nearly total loss of NEFL messenger RNA (mRNA), leading to the complete absence of NEFL protein in patient’s cultured neurons. Yet the cultured neurons were able to differentiate and form neuronal networks and neuro- filaments. Single-neuron gene expression fingerprinting pinpointed NEFL as the most down- regulated gene in the patient neurons and provided data of intermediate filament transcript abundancy and dynamics in cultured neurons. Blocking of nonsense-mediated decay partially rescued the loss of NEFL mRNA. Conclusions The strict neuronal specificity of neurofilament has hindered the mechanistic studies of re- cessive NEFL nonsense mutations. Here, we show that such mutation leads to the absence of NEFL, causing childhood-onset neuropathy through a loss-of-function mechanism.
    [Show full text]
  • Serum Albumin OS=Homo Sapiens
    Protein Name Cluster of Glial fibrillary acidic protein OS=Homo sapiens GN=GFAP PE=1 SV=1 (P14136) Serum albumin OS=Homo sapiens GN=ALB PE=1 SV=2 Cluster of Isoform 3 of Plectin OS=Homo sapiens GN=PLEC (Q15149-3) Cluster of Hemoglobin subunit beta OS=Homo sapiens GN=HBB PE=1 SV=2 (P68871) Vimentin OS=Homo sapiens GN=VIM PE=1 SV=4 Cluster of Tubulin beta-3 chain OS=Homo sapiens GN=TUBB3 PE=1 SV=2 (Q13509) Cluster of Actin, cytoplasmic 1 OS=Homo sapiens GN=ACTB PE=1 SV=1 (P60709) Cluster of Tubulin alpha-1B chain OS=Homo sapiens GN=TUBA1B PE=1 SV=1 (P68363) Cluster of Isoform 2 of Spectrin alpha chain, non-erythrocytic 1 OS=Homo sapiens GN=SPTAN1 (Q13813-2) Hemoglobin subunit alpha OS=Homo sapiens GN=HBA1 PE=1 SV=2 Cluster of Spectrin beta chain, non-erythrocytic 1 OS=Homo sapiens GN=SPTBN1 PE=1 SV=2 (Q01082) Cluster of Pyruvate kinase isozymes M1/M2 OS=Homo sapiens GN=PKM PE=1 SV=4 (P14618) Glyceraldehyde-3-phosphate dehydrogenase OS=Homo sapiens GN=GAPDH PE=1 SV=3 Clathrin heavy chain 1 OS=Homo sapiens GN=CLTC PE=1 SV=5 Filamin-A OS=Homo sapiens GN=FLNA PE=1 SV=4 Cytoplasmic dynein 1 heavy chain 1 OS=Homo sapiens GN=DYNC1H1 PE=1 SV=5 Cluster of ATPase, Na+/K+ transporting, alpha 2 (+) polypeptide OS=Homo sapiens GN=ATP1A2 PE=3 SV=1 (B1AKY9) Fibrinogen beta chain OS=Homo sapiens GN=FGB PE=1 SV=2 Fibrinogen alpha chain OS=Homo sapiens GN=FGA PE=1 SV=2 Dihydropyrimidinase-related protein 2 OS=Homo sapiens GN=DPYSL2 PE=1 SV=1 Cluster of Alpha-actinin-1 OS=Homo sapiens GN=ACTN1 PE=1 SV=2 (P12814) 60 kDa heat shock protein, mitochondrial OS=Homo
    [Show full text]
  • Spatial Distribution of Leading Pacemaker Sites in the Normal, Intact Rat Sinoa
    Supplementary Material Supplementary Figure 1: Spatial distribution of leading pacemaker sites in the normal, intact rat sinoatrial 5 nodes (SAN) plotted along a normalized y-axis between the superior vena cava (SVC) and inferior vena 6 cava (IVC) and a scaled x-axis in millimeters (n = 8). Colors correspond to treatment condition (black: 7 baseline, blue: 100 µM Acetylcholine (ACh), red: 500 nM Isoproterenol (ISO)). 1 Supplementary Figure 2: Spatial distribution of leading pacemaker sites before and after surgical 3 separation of the rat SAN (n = 5). Top: Intact SAN preparations with leading pacemaker sites plotted during 4 baseline conditions. Bottom: Surgically cut SAN preparations with leading pacemaker sites plotted during 5 baseline conditions (black) and exposure to pharmacological stimulation (blue: 100 µM ACh, red: 500 nM 6 ISO). 2 a &DUGLDFIoQChDQQHOV .FQM FOXVWHU &DFQDG &DFQDK *MD &DFQJ .FQLS .FQG .FQK .FQM &DFQDF &DFQE .FQM í $WSD .FQD .FQM í .FQN &DVT 5\U .FQM &DFQJ &DFQDG ,WSU 6FQD &DFQDG .FQQ &DFQDJ &DFQDG .FQD .FQT 6FQD 3OQ 6FQD +FQ *MD ,WSU 6FQE +FQ *MG .FQN .FQQ .FQN .FQD .FQE .FQQ +FQ &DFQDD &DFQE &DOP .FQM .FQD .FQN .FQG .FQN &DOP 6FQD .FQD 6FQE 6FQD 6FQD ,WSU +FQ 6FQD 5\U 6FQD 6FQE 6FQD .FQQ .FQH 6FQD &DFQE 6FQE .FQM FOXVWHU V6$1 L6$1 5$ /$ 3 b &DUGLDFReFHSWRUV $GUDF FOXVWHU $GUDD &DY &KUQE &KUP &KJD 0\O 3GHG &KUQD $GUE $GUDG &KUQE 5JV í 9LS $GUDE 7SP í 5JV 7QQF 3GHE 0\K $GUE *QDL $QN $GUDD $QN $QN &KUP $GUDE $NDS $WSE 5DPS &KUP 0\O &KUQD 6UF &KUQH $GUE &KUQD FOXVWHU V6$1 L6$1 5$ /$ 4 c 1HXURQDOPURWHLQV
    [Show full text]
  • Supplemental Text and Figures
    Supplemental Materials and Methods Experimental bone metastasis assay Primary PCa cells were sorted by GFP marker from mTmG+ tumors, and 105 cells in 20μL PBS were injected using Hamilton syringe into the tibia of 6-week old NSG mice. Mice were monitored biweekly for moribund signs for euthanasia and organ harvest. Noninvasive mouse and ex vivo imaging MRI imaging with Bruker ICON and fluorescence imaging of fresh organs with metastasis enumeration were recently described (Lu et al. 2017). Primary prostate cell sphere formation assay Isolate of primary cells from prostate, culture and counting of prostatospheres on Matrigel were performed as described (Lukacs et al. 2010). For organoid culture assay, we followed a published matrigel embedding method (Chua et al. 2014). RNA-Seq and differential gene expression Total RNA was isolated from prostate tumors using Direct-zol RNA MiniPrep Kit (Zymo Research) and processed for stranded total RNA-Seq using Illumina HiSeq 4000 at Sequencing and Microarray Facility at MD Anderson Cancer Center. The differential expression analysis was performed using the DESeq2 package of R. P-values obtained after multiple binomial tests were adjusted using BH method. Significant genes are defined by using a cut-off of 0.05 on the BH corrected p-value and an absolute log2 fold change value of at least 1.5. Histology and western blot H&E stain, immunohistochemical (IHC) and western blot were performed as previously described (Ding et al. 2011; Wang et al. 2016). Primary antibodies for IHC include Ki67 (Fisher, RM-9106-S1), cleaved caspase 3 (Cell Signaling Technology aka CST, 9661), cyclin D1 (Fisher, clone SP4), TGFBR2 (Abcam, ab61213), BMPR2 (Abcam, ab130206), AR (EMD Millipore, 06-680), phospho- Akt (CST, 4060), GFP (CST, 2956), E-Cadherin (CST, 14472).
    [Show full text]
  • Homer1 Promotes Dendritic Spine Growth Through Ankyrin-G and Its Loss Reshapes the Synaptic Proteome
    Molecular Psychiatry (2021) 26:1775–1789 https://doi.org/10.1038/s41380-020-00991-1 ARTICLE Homer1 promotes dendritic spine growth through ankyrin-G and its loss reshapes the synaptic proteome 1 1 2 1,5 2 Sehyoun Yoon ● Nicolas H. Piguel ● Natalia Khalatyan ● Leonardo E. Dionisio ● Jeffrey N. Savas ● Peter Penzes 1,3,4 Received: 22 February 2020 / Revised: 24 November 2020 / Accepted: 7 December 2020 / Published online: 4 January 2021 © The Author(s), under exclusive licence to Springer Nature Limited part of Springer Nature 2021. This article is published with open access Abstract Homer1 is a synaptic scaffold protein that regulates glutamatergic synapses and spine morphogenesis. HOMER1 knockout (KO) mice show behavioral abnormalities related to psychiatric disorders, and HOMER1 has been associated with psychiatric disorders such as addiction, autism disorder (ASD), schizophrenia (SZ), and depression. However, the mechanisms by which it promotes spine stability and its global function in maintaining the synaptic proteome has not yet been fully investigated. Here, we used computational approaches to identify global functions for proteins containing the Homer1-interacting PPXXF motif within the postsynaptic compartment. Ankyrin-G was one of the most topologically important nodes in the postsynaptic peripheral 1234567890();,: 1234567890();,: membrane subnetwork, and we show that one of the PPXXF motifs, present in the postsynaptically-enriched 190 kDa isoform of ankyrin-G (ankyrin-G 190), is recognized by the EVH1 domain of Homer1. We use proximity ligation combined with super- resolution microscopy to map the interaction of ankyrin-G and Homer1 to distinct nanodomains within the spine head and correlate them with spine head size.
    [Show full text]
  • Supplementary Table 1. the List of Proteins with at Least 2 Unique
    Supplementary table 1. The list of proteins with at least 2 unique peptides identified in 3D cultured keratinocytes exposed to UVA (30 J/cm2) or UVB irradiation (60 mJ/cm2) and treated with treated with rutin [25 µM] or/and ascorbic acid [100 µM]. Nr Accession Description 1 A0A024QZN4 Vinculin 2 A0A024QZN9 Voltage-dependent anion channel 2 3 A0A024QZV0 HCG1811539 4 A0A024QZX3 Serpin peptidase inhibitor 5 A0A024QZZ7 Histone H2B 6 A0A024R1A3 Ubiquitin-activating enzyme E1 7 A0A024R1K7 Tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein 8 A0A024R280 Phosphoserine aminotransferase 1 9 A0A024R2Q4 Ribosomal protein L15 10 A0A024R321 Filamin B 11 A0A024R382 CNDP dipeptidase 2 12 A0A024R3V9 HCG37498 13 A0A024R3X7 Heat shock 10kDa protein 1 (Chaperonin 10) 14 A0A024R408 Actin related protein 2/3 complex, subunit 2, 15 A0A024R4U3 Tubulin tyrosine ligase-like family 16 A0A024R592 Glucosidase 17 A0A024R5Z8 RAB11A, member RAS oncogene family 18 A0A024R652 Methylenetetrahydrofolate dehydrogenase 19 A0A024R6C9 Dihydrolipoamide S-succinyltransferase 20 A0A024R6D4 Enhancer of rudimentary homolog 21 A0A024R7F7 Transportin 2 22 A0A024R7T3 Heterogeneous nuclear ribonucleoprotein F 23 A0A024R814 Ribosomal protein L7 24 A0A024R872 Chromosome 9 open reading frame 88 25 A0A024R895 SET translocation 26 A0A024R8W0 DEAD (Asp-Glu-Ala-Asp) box polypeptide 48 27 A0A024R9E2 Poly(A) binding protein, cytoplasmic 1 28 A0A024RA28 Heterogeneous nuclear ribonucleoprotein A2/B1 29 A0A024RA52 Proteasome subunit alpha 30 A0A024RAE4 Cell division cycle 42 31
    [Show full text]
  • New Partners Identified by Mass Spectrometry Assay Reveal Functions of NCAM2 in Neural Cytoskeleton Organization
    International Journal of Molecular Sciences Article New Partners Identified by Mass Spectrometry Assay Reveal Functions of NCAM2 in Neural Cytoskeleton Organization Antoni Parcerisas 1,2,3,*,† , Alba Ortega-Gascó 1,2,† , Marc Hernaiz-Llorens 1,2 , Maria Antonia Odena 4, Fausto Ulloa 1,2, Eliandre de Oliveira 4, Miquel Bosch 3 , Lluís Pujadas 1,2 and Eduardo Soriano 1,2,* 1 Department of Cell Biology, Physiology and Immunology, University of Barcelona and Institute of Neurosciences, 08028 Barcelona, Spain; [email protected] (A.O.-G.); [email protected] (M.H.-L.); [email protected] (F.U.); [email protected] (L.P.) 2 Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (CIBERNED), 28031 Madrid, Spain 3 Department of Basic Sciences, Universitat Internacional de Catalunya, 08195 Sant Cugat del Vallès, Spain; [email protected] 4 Plataforma de Proteòmica, Parc Científic de Barcelona (PCB), 08028 Barcelona, Spain; [email protected] (M.A.O.); [email protected] (E.d.O.) * Correspondence: [email protected] (A.P.); [email protected] (E.S.) † A.P. and A.O.-G. contributed equally. Abstract: Neuronal cell adhesion molecule 2 (NCAM2) is a membrane protein with an important role in the morphological development of neurons. In the cortex and the hippocampus, NCAM2 is essential for proper neuronal differentiation, dendritic and axonal outgrowth and synapse forma- tion. However, little is known about NCAM2 functional mechanisms and its interactive partners Citation: Parcerisas, A.; during brain development. Here we used mass spectrometry to study the molecular interactome Ortega-Gascó, A.; Hernaiz-Llorens, of NCAM2 in the second postnatal week of the mouse cerebral cortex.
    [Show full text]
  • Neurofilament (NEFM) Mouse Monoclonal Antibody [Clone ID: OTI2G3] Product Data
    OriGene Technologies, Inc. 9620 Medical Center Drive, Ste 200 Rockville, MD 20850, US Phone: +1-888-267-4436 [email protected] EU: [email protected] CN: [email protected] Product datasheet for TA805823 Neurofilament (NEFM) Mouse Monoclonal Antibody [Clone ID: OTI2G3] Product data: Product Type: Primary Antibodies Clone Name: OTI2G3 Applications: IHC, WB Recommended Dilution: WB 1:2000, IHC 1:150 Reactivity: Human Host: Mouse Isotype: IgG1 Clonality: Monoclonal Immunogen: Human recombinant protein fragment corresponding to amino acids 413-773 of human NEFM(NP_005373) produced in E.coli. Formulation: PBS (PH 7.3) containing 1% BSA, 50% glycerol and 0.02% sodium azide. Concentration: 1 mg/ml Purification: Purified from mouse ascites fluids or tissue culture supernatant by affinity chromatography (protein A/G) Conjugation: Unconjugated Storage: Store at -20°C as received. Stability: Stable for 12 months from date of receipt. Predicted Protein Size: 102.3 kDa Gene Name: neurofilament, medium polypeptide Database Link: NP_005373 Entrez Gene 4741 Human P07197 Background: Neurofilaments are type IV intermediate filament heteropolymers composed of light, medium, and heavy chains. Neurofilaments comprise the axoskeleton and functionally maintain neuronal caliber. They may also play a role in intracellular transport to axons and dendrites. This gene encodes the medium neurofilament protein. This protein is commonly used as a biomarker of neuronal damage. Alternative splicing results in multiple transcript variants encoding distinct
    [Show full text]
  • Supplementary Information
    Osa et al Supplementary Information Clinical implications of monitoring nivolumab immunokinetics in previously treated non– small cell lung cancer patients Akio Osa, Takeshi Uenami, Shohei Koyama, Kosuke Fujimoto, Daisuke Okuzaki, Takayuki Takimoto, Haruhiko Hirata, Yukihiro Yano, Soichiro Yokota, Yuhei Kinehara, Yujiro Naito, Tomoyuki Otsuka, Masaki Kanazu, Muneyoshi Kuroyama, Masanari Hamaguchi, Taro Koba, Yu Futami, Mikako Ishijima, Yasuhiko Suga, Yuki Akazawa, Hirotomo Machiyama, Kota Iwahori, Hyota Takamatsu, Izumi Nagatomo, Yoshito Takeda, Hiroshi Kida, Esra A. Akbay, Peter S. Hammerman, Kwok-kin Wong, Glenn Dranoff, Masahide Mori, Takashi Kijima, Atsushi Kumanogoh Supplemental Figures 1 – 8 1 Osa et al Supplemental Figure 1. The frequency of nivolumab-bound T cells was maintained in patients who continued treatment. Nivolumab binding in CD8 and CD4 T cells was analyzed at two follow-up points, as indicated, in fresh peripheral blood from three representative cases from protocol 1 that continued treatment. 2 Osa et al Supplemental Figure 2. Long-term follow-up of nivolumab binding to T cells from fresh whole blood. Nivolumab binding was followed up in fresh peripheral blood from an additional case, Pt.7. 3 Osa et al Supplemental Figure 3. Long-term duration of nivolumab binding is due to sustained circulation of residual nivolumab in plasma. (A) PBMCs acquired from Pt.8 and 9 at pretreatment (pre PBMCs) and after a single dose (post 1 PBMCs) were cultured in regular medium without nivolumab (top and middle). Pre PBMCs were also incubated with 10 µg/ml nivolumab in vitro before the cultures were started (bottom). Nivolumab binding status was monitored at the indicated time points.
    [Show full text]
  • Identification, Quantification and Investigation of Anti-Inflammatory Effects of Echinacea Purpurea Constituents
    KANDHI, VAMSIKRISHNA, Ph.D. Identification, Quantification and Investigation of Anti-inflammatory Effects of Echinacea purpurea Constituents. (2012) Directed by Dr. Nadja B. Cech. 263 pp. Echinacea is among the top selling botanical medicines in the United States. Clinical trials have yielded contradictory reports on the efficacy of Echinacea preparations for treatment of colds, influenza, and inflammation, perhaps due to inconsistency in quality control and lack of information as to which constituents are responsible for their activity. Our overall goal with this research was to develop and apply mass spectrometry techniques to better understand the relationship between chemical composition of Echinacea extracts and their biological activity. This objective was achieved through three separate aims. (1) To determine whether in vitro anti-inflammatory activity of Echinacea purpurea extracts could be correlated with the presence of the alkylamides or caffeic acid derivatives, which have previously been reported to be bioactive. The ability of these extracts to inhibit production of TNF-alpha and PGE2 from influenza A-infected RAW 264.7 macrophage-like cells was assessed. Chemical analysis of the extracts revealed variations in levels of alkylamides, caftaric acid, and cichoric acid. The biological activity of these extracts, however, did not correlate with concentrations of any of these compounds. (2) To investigate the effects of Echinacea purpurea preparations and their constituents on protein expression by immune cells in vitro. Stable isotope labeling with amino acid in cell culture (SILAC), a quantitative proteomics approach, was employed to measure changes in protein expression by activated RAW 264.7 macrophage- type cells exposed to an Echinacea purpurea extract and several of its constituents.
    [Show full text]
  • Genome-Wide Screening Identifies Genes and Biological Processes
    Louisiana State University LSU Digital Commons LSU Doctoral Dissertations Graduate School 10-12-2018 Genome-Wide Screening Identifies Genes and Biological Processes Implicated in Chemoresistance and Oncogene-Induced Apoptosis Tengyu Ko Louisiana State University and Agricultural and Mechanical College, [email protected] Follow this and additional works at: https://digitalcommons.lsu.edu/gradschool_dissertations Part of the Cancer Biology Commons, Cell Biology Commons, and the Genomics Commons Recommended Citation Ko, Tengyu, "Genome-Wide Screening Identifies Genes and Biological Processes Implicated in Chemoresistance and Oncogene- Induced Apoptosis" (2018). LSU Doctoral Dissertations. 4715. https://digitalcommons.lsu.edu/gradschool_dissertations/4715 This Dissertation is brought to you for free and open access by the Graduate School at LSU Digital Commons. It has been accepted for inclusion in LSU Doctoral Dissertations by an authorized graduate school editor of LSU Digital Commons. For more information, please [email protected]. GENOME-WIDE SCREENING IDENTIFIES GENES AND BIOLOGICAL PROCESSES IMPLICATED IN CHEMORESISTANCE AND ONCOGENE- INDUCED APOPTOSIS A Dissertation Submitted to the Graduate Faculty of the Louisiana State University and Agricultural and Mechanical College in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Biomedical and Veterinary Medical Sciences through the Department of Comparative Biomedical Sciences by Tengyu Ko B.S., University of California, Santa Barbara 2010 December 2018 ACKNOWLEDGEMENTS I would like to express my sincerest gratitude to my major supervisor Dr. Shisheng Li for giving me the opportunity to join his team and the freedom to pursue projects. I appreciate all of his thoughts and efforts. Truly, none of these findings would be possible without his supervisions, supports, insightful discussions, and patience.
    [Show full text]