DOCSLIB.ORG

Characterization of Gene-Activated Human Acid-Β-Glucosidase: Crystal Structure, Glycan Composition, and Internalization Into Macrophages

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Characterization of Gene-Activated Human Acid-Β-Glucosidase: Crystal Structure, Glycan Composition, and Internalization Into Macrophages Glycobiology vol. 20 no. 1 pp. 24–32, 2010 doi:10.1093/glycob/cwp138 Advance Access publication on September 9, 2009 Characterization of gene-activated human acid-β-glucosidase: Crystal structure, glycan composition, and internalization into macrophages Boris Brumshtein2,3, Paul Salinas4, Brian Peterson4,Victor Futerman and Zimran 2006). The most common treatment for Chan4, Israel Silman2, Joel L Sussman3, Philip J Savickas4, Gaucher disease is enzyme replacement therapy (ERT), in which Gregory S Robinson4, and Anthony H Futerman1,5 defective GlcCerase is supplemented with an active enzyme. ERT using imiglucerase, a recombinant analog of human Glc- 2Department of Neurobiology; 3Department of Structural Biology, Weizmann Institute of Science, Rehovot 76100, Israel; 4Shire Human Genetic Therapies, Cerase expressed in Chinese hamster ovary (CHO) cells has Inc., Cambridge, MA, USA; and 5Department of Biological Chemistry, been available for ∼15 years. After expression and purifica- Weizmann Institute of Science, Rehovot 76100, Israel tion, imiglucerase is modified by exo-glycosidase treatment Received on June 1, 2009; revised on August 30, 2009; accepted on September (Friedman and Hayes 1996) to expose the core mannose 1, 2009 residues that can be recognized by macrophages. Glycan re- modeling greatly improves targeting to and internalization by Gaucher disease, the most common lysosomal storage dis- macrophages, the main cell type affected in Gaucher disease ease, can be treated with enzyme replacement therapy (Futerman and Zimran 2006). An alternative means of pro- (ERT), in which defective acid-β-glucosidase (GlcCerase) ducing GlcCerase (prGCD) in transgenic carrot root cells has is supplemented by a recombinant, active enzyme. The been developed (Aviezer et al. 2009). The X-ray structures of X-ray structures of recombinant GlcCerase produced in imiglucerase and prGCD have been previously reported (Dvir Chinese hamster ovary cells (imiglucerase, CerezymeR ) and et al. 2003; Shaaltiel et al. 2007). in transgenic carrot cells (prGCD) have been previously In the current study, we have used gene activation in solved. We now describe the structure and characteristics a well-characterized, continuous human cell line to pro- of a novel form of GlcCerase under investigation for the duce gene-activated human acid-β-glucocerebrosidase (ve- treatment of Gaucher disease, Gene-ActivatedTM human laglucerase alfa). Gene activation refers to targeted recombi- GlcCerase (velaglucerase alfa). In contrast to imiglucerase nation with a promoter that activates the endogenous GlcCerase and prGCD, velaglucerase alfa contains the native human gene in the selected human cell line. Velaglucerase alfa is se- enzyme sequence. All three GlcCerases consist of three creted as a monomeric glycoprotein of approximately 63 kDa domains, with the active site located in domain III. The and is composed of 497 amino acids with a sequence identical distances between the carboxylic oxygens of the catalytic to that of the natural human protein (Zimran et al. 2007). Glyco- residues, E340 and E235, are consistent with distances pro- sylation of velaglucerase alfa is altered by using kifunensine, a posed for acid–base hydrolysis. Kinetic parameters (Km and mannosidase I inhibitor, during cell culture, which results in the Vmax) of velaglucerase alfa and imiglucerase, as well as their secretion of a protein containing predominantly high-mannose specific activities, are similar. However, analysis of glycosy- type glycans (Elbein et al. 1990). lation patterns shows that velaglucerase alfa displays dis- Herein we describe the crystal structure of velaglucerase alfa, tinctly different structures from imiglucerase and prGCD. using a preparation that had been partially deglycosylated, and The predominant glycan on velaglucerase alfa is a high- show that it is similar to that of imiglucerase (Dvir et al. 2003) mannose type, with nine mannose units, while imiglucerase and prGCD (Shaaltiel et al. 2007). Velaglucerase alfa differs contains a chitobiose tri-mannosyl core glycan with fuco- from imiglucerase and prGCD as the latter two enzymes contain sylation. These differences in glycosylation affect cellular a mutation at residue 495 (an Arg to His substitution: R495H), internalization; the rate of velaglucerase alfa internaliza- and prGCD contains seven additional residues at the C terminus tion into human macrophages is at least 2-fold greater than (DLLVDTM) and two additional residues at the N terminus that of imiglucerase. (EF). Moreover, the kinetic parameters and specific activity of velaglucerase alfa are very similar to those of imiglucerase. We Keywords: Gaucher disease/gene activation/ also compare the glycosylation patterns of velaglucerase alfa glucocerebrosidase/glycans/mannose-6-phosphate receptor/ and imiglucerase by use of LC-MS and assess the impact of the site-specific glycosylation/X-ray structure different glycosylation patterns by analyzing internalization in human macrophages. Introduction Results and discussion Gaucher disease is caused by mutations in the gene encod- ing the lysosomal enzyme, acid-β-glucosidase (glucocerebrosi- X-ray structure dase, GlcCerase, E.C. 3.2.1.45) (Beutler and Grabowski 2001; Diffraction-quality crystals of velaglucerase alfa were obtained after partial deglycosylation using N-glycosidase F, by a proce- 1To whom correspondence should be addressed: Tel: +972-8-9342704; dure similar to that previously described for imiglucerase (Dvir Fax: +972-8-9344112; e-mail: [email protected] et al. 2003). Velaglucerase alfa crystallized in the same space c 2009 The Author(s). Published by Oxford University Press. All rights reserved. This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5),which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. 24 Characterization of gene-activated human acid-β-glucosidase Table I. Data collection and refinement statistics B(<0.3 A)˚ shows that they are virtually identical. A compari- Velaglucerase alfa son of the structures of imiglucerase, prGCD, and velaglucerase alfa demonstrates that these structures are very similar, with an Data collection RMSD of 0.35–0.46 A˚ (Table II). Space group C2221 Velaglucerase alfa thus consists of three noncontiguous do- Cell dimensions mains, with the catalytic site located in domain III (residues 76– a, b, c (A)˚ 109.37, 285.55, 91.69 ◦ β α abg ( ) 90.00, 90.00, 90.00 381 and 416–430), which is a ( / )8 (TIM) barrel (Figure 1). Resolution (A)˚ 19.9–2.7 (2.75–2.70)a A more detailed analysis of the active site reveals that it is Rsym(%) 15.7 (51.0) virtually identical to that of imiglucerase (Figure 2), with the dis- I/s<I> 14.4 (4.6) tances between the carboxylic oxygens of the catalytic residues, Completeness (%) 100 (100) ˚ ˚ Redundancy 7.5 (7.6) E340 and E235 (5.2 A in molecule A and 5.1 A in molecule B), Refinement similar to those obtained previously (Brumshtein et al. 2006) Resolution (A)˚ 19.9–2.7 and in agreement with the distances proposed for acid–base hy- Number of reflections 39,776 drolysis (Davies and Henrissat 1995). Moreover, the three loops Rwork/Rfree 17.3/23.4 (loop 1, residues 345–350; loop 2, residues 393–399; and loop Rms deviations 3, residues 312–319) observed in previous structures (reviewed Bond lengths (A)˚ 0.012 Bond angles (◦) 1.486 in Kacher et al. (2008)) are also seen in velaglucerase alfa. Number of refined atoms Similarly to prGCD (Shaaltiel et al. 2007), loops 2 and 3 show Protein 7871 differences in their backbone angles and side chain orientations Carbohydrates 70 in the two molecules of the asymmetric unit, whereas loop 1, Ions 90 Solvent 326 since it makes crystal contacts, exhibits less pronounced con- Ramachandran outliers (%) 0.4 formational changes (Figure 2). In the case of loop 3, a helical conformation is seen in molecule B, whereas a coiled confor- aThe highest resolution shell is shown in parentheses. mation is seen in molecule A (Figure 3), as previously reported for imiglucerase (Brumshtein et al. 2006). Although the crystal was cryo-protected with 25% ethylene glycol, we did not detect group, C2221, as imiglucerase (Table I), and unit cell parameters were similar to the previously published GlcCerase structures any ethylene glycol molecules in the electron density map. (Dvir et al. 2003; Premkumar et al. 2005; Brumshtein et Imiglucerase and prGCD both contain an Arg to His muta- al. 2006). The asymmetric unit contained two copies of ve- tion at residue 495, with H495 making an H-bond (2.6 A)˚ with laglucerase alfa, designated as molecules A and B. The root the peptide carbonyl of F31. In contrast, velaglucerase alfa con- mean square deviation (RMSD) value between molecules A and tains a sequence identical to that of the natural human enzyme, Table II. RMS deviations of velaglucerase alfa compared to imiglucerase and prGCD. RMS deviations (A)˚ are shown for each of the two copies of the molecules in the asymmetric unit and were calculated using PyMol (www.pymol.org). The PDB codes for imiglucerase and pr-GlcCerase are 2J25 and 2V3F, respectively Imiglucerase-A Imiglucerase–B prGCD-A prGCD-B Velaglucerase alfa-A 0.39 0.35 0.36 0.40 Velaglucerase alfa-B 0.38 0.43 0.46 0.46 Fig. 1. Comparison of the crystal structures of velaglucerase alfa and imiglucerase. The three domains of the enzymes are colored pink (domain I, residues 1–29 and 383–414), blue (domain II, residues 30–75 and 431–497), and gray (domain III, residues 76–382 and 415–430). 25 B Brumshtein et al. Fig. 2. Active site of velaglucerase alfa. Stereo representation of an overlay of the active sites of imiglucerase (blue and magenta) and velaglucerase alfa (yellow and green). Catalytic residues are shown as red sticks. Loops near the entrance to the active site are indicated (L1, loop 1; L2, loop 2; L3, loop 3). with an Arg at residue 495, which does not make a similar Kinetic analysis H-bond.
Load more

Recommended publications
  • Bacteria Belonging to Pseudomonas Typographi Sp. Nov. from the Bark Beetle Ips Typographus Have Genomic Potential to Aid in the Host Ecology
    insects Article Bacteria Belonging to Pseudomonas typographi sp. nov. from the Bark Beetle Ips typographus Have Genomic Potential to Aid in the Host Ecology Ezequiel Peral-Aranega 1,2 , Zaki Saati-Santamaría 1,2 , Miroslav Kolaˇrik 3,4, Raúl Rivas 1,2,5 and Paula García-Fraile 1,2,4,5,* 1 Microbiology and Genetics Department, University of Salamanca, 37007 Salamanca, Spain; [email protected] (E.P.-A.); [email protected] (Z.S.-S.); [email protected] (R.R.) 2 Spanish-Portuguese Institute for Agricultural Research (CIALE), 37185 Salamanca, Spain 3 Department of Botany, Faculty of Science, Charles University, Benátská 2, 128 01 Prague, Czech Republic; [email protected] 4 Laboratory of Fungal Genetics and Metabolism, Institute of Microbiology of the Academy of Sciences of the Czech Republic, 142 20 Prague, Czech Republic 5 Associated Research Unit of Plant-Microorganism Interaction, University of Salamanca-IRNASA-CSIC, 37008 Salamanca, Spain * Correspondence: [email protected] Received: 4 July 2020; Accepted: 1 September 2020; Published: 3 September 2020 Simple Summary: European Bark Beetle (Ips typographus) is a pest that affects dead and weakened spruce trees. Under certain environmental conditions, it has massive outbreaks, resulting in attacks of healthy trees, becoming a forest pest. It has been proposed that the bark beetle’s microbiome plays a key role in the insect’s ecology, providing nutrients, inhibiting pathogens, and degrading tree defense compounds, among other probable traits. During a study of bacterial associates from I. typographus, we isolated three strains identified as Pseudomonas from different beetle life stages. In this work, we aimed to reveal the taxonomic status of these bacterial strains and to sequence and annotate their genomes to mine possible traits related to a role within the bark beetle holobiont.
    [Show full text]
  • Intravenous Enzyme Replacement Therapy (ERT) for Gaucher Disease
    UnitedHealthcare® Commercial Medical Benefit Drug Policy Intravenous Enzyme Replacement Therapy (ERT) for Gaucher Disease Policy Number: 2021D0048K Effective Date: April 1, 2021 Instructions for Use Table of Contents Page Related Commercial Policy Coverage Rationale ....................................................................... 1 • Provider Administered Drugs – Site of Care Applicable Codes .......................................................................... 3 Background.................................................................................... 3 Community Plan Policy Benefit Considerations .................................................................. 3 • Intravenous Enzyme Replacement Therapy (ERT) Clinical Evidence ........................................................................... 4 for Gaucher Disease U.S. Food and Drug Administration ............................................. 6 Centers for Medicare and Medicaid Services ............................. 6 References ..................................................................................... 7 Policy History/Revision Information ............................................. 8 Instructions for Use ....................................................................... 8 Coverage Rationale See Benefit Considerations This policy refers to the following drug products, all of which are intravenous enzyme replacement therapies used in the treatment of Gaucher disease: Cerezyme® (imiglucerase) Elelyso® (taliglucerase) VPRIV® (velaglucerase)
    [Show full text]
  • United States Patent (19) 11 Patent Number: 5,981,835 Austin-Phillips Et Al
    USOO598.1835A United States Patent (19) 11 Patent Number: 5,981,835 Austin-Phillips et al. (45) Date of Patent: Nov. 9, 1999 54) TRANSGENIC PLANTS AS AN Brown and Atanassov (1985), Role of genetic background in ALTERNATIVE SOURCE OF Somatic embryogenesis in Medicago. Plant Cell Tissue LIGNOCELLULOSC-DEGRADING Organ Culture 4:107-114. ENZYMES Carrer et al. (1993), Kanamycin resistance as a Selectable marker for plastid transformation in tobacco. Mol. Gen. 75 Inventors: Sandra Austin-Phillips; Richard R. Genet. 241:49-56. Burgess, both of Madison; Thomas L. Castillo et al. (1994), Rapid production of fertile transgenic German, Hollandale; Thomas plants of Rye. Bio/Technology 12:1366–1371. Ziegelhoffer, Madison, all of Wis. Comai et al. (1990), Novel and useful properties of a chimeric plant promoter combining CaMV 35S and MAS 73 Assignee: Wisconsin Alumni Research elements. Plant Mol. Biol. 15:373-381. Foundation, Madison, Wis. Coughlan, M.P. (1988), Staining Techniques for the Detec tion of the Individual Components of Cellulolytic Enzyme 21 Appl. No.: 08/883,495 Systems. Methods in Enzymology 160:135-144. de Castro Silva Filho et al. (1996), Mitochondrial and 22 Filed: Jun. 26, 1997 chloroplast targeting Sequences in tandem modify protein import specificity in plant organelles. Plant Mol. Biol. Related U.S. Application Data 30:769-78O. 60 Provisional application No. 60/028,718, Oct. 17, 1996. Divne et al. (1994), The three-dimensional crystal structure 51 Int. Cl. ............................. C12N 15/82; C12N 5/04; of the catalytic core of cellobiohydrolase I from Tricho AO1H 5/00 derma reesei. Science 265:524-528.
    [Show full text]
  • Are Glucosylceramide-Related Sphingolipids Involved in the Increased Risk for Cancer in Gaucher Disease Patients? Review and Hypotheses
    cancers Review Are Glucosylceramide-Related Sphingolipids Involved in the Increased Risk for Cancer in Gaucher Disease Patients? Review and Hypotheses 1,2, 1,3, 1 1,2 Patricia Dubot y , Leonardo Astudillo y, Nicole Therville , Frédérique Sabourdy , Jérôme Stirnemann 4 , Thierry Levade 1,2,* and Nathalie Andrieu-Abadie 1,* 1 INSERM UMR1037, CRCT (Cancer Research Center of Toulouse), and Université Paul Sabatier, 31037 Toulouse, France; [email protected] (P.D.); [email protected] (L.A.); [email protected] (N.T.); [email protected] (F.S.) 2 Laboratoire de Biochimie Métabolique, Centre de Référence en Maladies Héréditaires du Métabolisme, Institut Fédératif de Biologie, CHU de Toulouse, 31059 Toulouse, France 3 Service de Médecine Interne, CHU de Toulouse, 31059 Toulouse, France 4 Service de Médecine Interne Générale, Hôpitaux Universitaires de Genève, CH-1211 Geneva, Switzerland; [email protected] * Correspondence: [email protected] (T.L.); [email protected] (N.A.-A.) These authors contributed equally to this work. y Received: 28 November 2019; Accepted: 14 February 2020; Published: 18 February 2020 Abstract: The roles of ceramide and its catabolites, i.e., sphingosine and sphingosine 1-phosphate, in the development of malignancies and the response to anticancer regimens have been extensively described. Moreover, an abundant literature points to the effects of glucosylceramide synthase, the mammalian enzyme that converts ceramide to β-glucosylceramide, in protecting tumor cells from chemotherapy. Much less is known about the contribution of β-glucosylceramide and its breakdown products in cancer progression. In this chapter, we first review published and personal clinical observations that report on the increased risk of developing cancers in patients affected with Gaucher disease, an inborn disorder characterized by defective lysosomal degradation of β-glucosylceramide.
    [Show full text]
  • Vpriv-Pm-En.Pdf
    PRODUCT MONOGRAPH INCLUDING PATIENT MEDICATION INFORMATION PrVPRIV® velaglucerase alfa Powder for Solution for Injection 400 U/vial, Intravenous Enzyme Replacement Therapy ATC code: A16AB10 Takeda Canada Inc. Date of Initial Approval: 22 Adelaide Street West, Suite 3800 October 1, 2010 Toronto Ontario M5H 4E3 Date of Revision: December 21, 2020 Submission Control No: 241844 VPRIV® and the VPRIV Logo® are registered trademarks of Shire Human Genetic Therapies, Inc. TAKEDA™ and the TAKEDA Logo® are trademarks of Takeda Pharmaceutical Company Limited, used under license. Page 1 of 24 RECENT MAJOR LABEL CHANGES Warnings and Precautions, Immunogenicity (8) 9/2020 Warnings and Precautions, Infusion-Related Reactions (8) 9/2020 Warnings and Precautions, Breast-feeding (8.1.2) 9/2020 TABLE OF CONTENTS PART I: HEALTH PROFESSIONAL INFORMATION ................................................................. 4 1 INDICATIONS .................................................................................................................4 2 CONTRAINDICATIONS ..................................................................................................4 4 DOSAGE AND ADMINISTRATION ................................................................................ 4 4.1 Dosing Considerations ..........................................................................................4 4.2 Recommended Dose and Dosage Adjustment ....................................................... 4 4.3 Administration .......................................................................................................4
    [Show full text]
  • Glucocerebrosidase Mutations and Synucleinopathies. Potential Role of Sterylglucosides and Relevance of Studying Both GBA1 and GBA2 Genes
    MINI REVIEW published: 28 June 2018 doi: 10.3389/fnana.2018.00052 Glucocerebrosidase Mutations and Synucleinopathies. Potential Role of Sterylglucosides and Relevance of Studying Both GBA1 and GBA2 Genes Rafael Franco 1,2*†, Juan A. Sánchez-Arias 3†, Gemma Navarro 1,2,4 and José L. Lanciego 2,3,5* 1Department of Biochemistry and Molecular Biomedicine, School of Biology, University of Barcelona, Barcelona, Spain, 2Centro de Investigación Biomédica en Red de Enfermedades Neurodegenerativas (CiberNed), Instituto de Salud Carlos III, Madrid, Spain, 3Department of Neuroscience, Centro de Investigación Médica Aplicada (CIMA), University of Navarra, Pamplona, Spain, 4Department of Biochemistry and Physiology, School of Pharmacy, University of Barcelona, Barcelona, Spain, 5Department of Neuroscience, Instituto de Investigación Sanitaria de Navarra (IdiSNA), Pamplona, Spain Gaucher’s disease (GD) is the most prevalent lysosomal storage disorder. GD is caused Edited by: by homozygous mutations of the GBA1 gene, which codes for beta-glucocerebrosidase Francesco Fornai, (GCase). Although GD primarily affects peripheral tissues, the presence of neurological Università degli Studi di Pisa, Italy symptoms has been reported in several GD subtypes. GBA1 mutations have Reviewed by: Rosario Moratalla, recently deserved increased attention upon the demonstration that both homo- and Consejo Superior de Investigaciones heterozygous GBA1 mutations represent the most important genetic risk factor for the Científicas (CSIC), Spain Fabrizio Michetti, appearance of synucleinopathies like Parkinson’s disease (PD) and dementia with Lewy Università Cattolica del Sacro Cuore, bodies (LBD). Although reduced GCase activity leads to alpha-synuclein aggregation, Italy the mechanisms sustaining a role for GCase in alpha-synuclein homeostasis still *Correspondence: Rafael Franco remain largely unknown.
    [Show full text]
  • Draft Genome of Thermomonospora Sp. CIT 1 (Thermomonosporaceae) and in Silico Evidence of Its Functional Role in Filter Cake Biomass Deconstruction
    1 Genetics and Molecular Biology Suplementary material to: Draft genome of Thermomonospora sp. CIT 1 (Thermomonosporaceae) and in silico evidence of its functional role in filter cake biomass deconstruction Table S3 - Identifications of enzymes with activity on carbohydrate structures present in the draft genome CIT 1 recovered from metagenomic sequencing of filter cake. Predictions were performed with dbCAN online, following for blastp confirmation against the non-redundant NCBI protein database of the occurrence of predicted protein-like sequence deposition. Search results for dbCAN conserved domains BLASTP RESULTS ORF CAZy QUERY E- IDENTIT LENGTH ID CAZy ENZYME NAME (NCBI) ACCESSION FAMILY COVER VALUE Y 747 AA2.hmm peroxidase (EC 1.11.1.-) catalase/peroxidase HPI 100 0.00E+00 100 WP_012852289 glucose-methanol-choline 786 AA3.hmm glucose-methanol-choline (GMC) 100 0.00E+00 100 ACY97563 oxidoreductase mycofactocin system GMC family 520 AA3.hmm glucose-methanol-choline (GMC) 99 0.00E+00 99 WP_012852714 oxidoreductase MftG 582 AA3.hmm glucose-methanol-choline (GMC) GMC family oxidoreductase 100 0.00E+00 99 WP_012854558 531 AA3_2.hmm glucose-methanol-choline (GMC) choline dehydrogenase 100 0.00E+00 100 WP_012854223 533 AA4.hmm vanillyl-alcohol oxidase (EC 1.1.3.38) FAD-binding oxidoreductase 99 0.00E+00 99 WP_012852156 NAD(P)H:quinone oxidoreductase 4.00E- 209 AA6.hmm 1,4-benzoquinone reductase (EC. 1.6.5.6) 100 100 WP_012850533 type IV 152 187 AA6.hmm 1,4-benzoquinone reductase (EC. 1.6.5.6) NAD(P)H-dependent oxidoreductase 100 9.00E-86 69 WP_067443322 6.00E- 152 AA6.hmm 1,4-benzoquinone reductase (EC.
    [Show full text]
  • Estonian Statistics on Medicines 2016 1/41
    Estonian Statistics on Medicines 2016 ATC code ATC group / Active substance (rout of admin.) Quantity sold Unit DDD Unit DDD/1000/ day A ALIMENTARY TRACT AND METABOLISM 167,8985 A01 STOMATOLOGICAL PREPARATIONS 0,0738 A01A STOMATOLOGICAL PREPARATIONS 0,0738 A01AB Antiinfectives and antiseptics for local oral treatment 0,0738 A01AB09 Miconazole (O) 7088 g 0,2 g 0,0738 A01AB12 Hexetidine (O) 1951200 ml A01AB81 Neomycin+ Benzocaine (dental) 30200 pieces A01AB82 Demeclocycline+ Triamcinolone (dental) 680 g A01AC Corticosteroids for local oral treatment A01AC81 Dexamethasone+ Thymol (dental) 3094 ml A01AD Other agents for local oral treatment A01AD80 Lidocaine+ Cetylpyridinium chloride (gingival) 227150 g A01AD81 Lidocaine+ Cetrimide (O) 30900 g A01AD82 Choline salicylate (O) 864720 pieces A01AD83 Lidocaine+ Chamomille extract (O) 370080 g A01AD90 Lidocaine+ Paraformaldehyde (dental) 405 g A02 DRUGS FOR ACID RELATED DISORDERS 47,1312 A02A ANTACIDS 1,0133 Combinations and complexes of aluminium, calcium and A02AD 1,0133 magnesium compounds A02AD81 Aluminium hydroxide+ Magnesium hydroxide (O) 811120 pieces 10 pieces 0,1689 A02AD81 Aluminium hydroxide+ Magnesium hydroxide (O) 3101974 ml 50 ml 0,1292 A02AD83 Calcium carbonate+ Magnesium carbonate (O) 3434232 pieces 10 pieces 0,7152 DRUGS FOR PEPTIC ULCER AND GASTRO- A02B 46,1179 OESOPHAGEAL REFLUX DISEASE (GORD) A02BA H2-receptor antagonists 2,3855 A02BA02 Ranitidine (O) 340327,5 g 0,3 g 2,3624 A02BA02 Ranitidine (P) 3318,25 g 0,3 g 0,0230 A02BC Proton pump inhibitors 43,7324 A02BC01 Omeprazole
    [Show full text]
  • ZHOU-THESIS-2018.Pdf (2.307Mb)
    PHYLOGENY AND PREDICTED FUNCTIONAL CAPABILITIES OF A SULFUR- OXIDIZING AND DENITRIFYING CLADE OF BACTEROIDETES FROM SULFIDIC ENVIRONMENTS A Thesis by KECEN ZHOU Submitted to the Office of Graduate and Professional Studies of Texas A&M University in partial fulfillment of the requirements for the degree of MASTER OF SCIENCE Chair of Committee, Jason B. Sylvan Co-Chair of Committee, Lisa Campbell Committee Member, Brendan E. Roark Head of Department, Shari Yvon-Lewis August 2018 Major Subject: Oceanography Copyright 2018 Kecen Zhou ABSTRACT Environments rich in sulfur compounds (sulfidic) are common in the ocean, and the ability to gain energy (dissimilatory) from sulfur redox reactions is widespread in bacteria. The Sulfiphilic Bacteroidetes (SB), have been found exclusively in sulfidic environments, but little is known about their metabolic potential and membership. The ability to perform dissimilatory sulfur redox would make them unique among Bacteroidetes, which are primarily known as heterotrophs that specialize in degrading complex organic molecules. Using 16S rRNA phylogeny and analysis of single amplified genomes (SAGs) from Saanich Inlet, a seasonally hypoxic basin, we elucidate the global distribution and potential metabolic capabilities of the SB clade. Phylogenetic analysis revealed that this clade was monophyletic and had a global distribution. It is hypothesized this clade combines heterotrophic amino acid and sugar uptake with denitrification and respiratory sulfur oxidation/polysulfide reduction. Putative genes for sulfur oxidation via polysulfide reductase (psr) were found in the combined genome, and phylogenetic analysis confirmed these genes were likely to be psrABC. A denitrification pathway was present and complete save for the absence of a gene catalyzing reduction of NO to N2O.
    [Show full text]
  • Isolation and Characterization of Cellulase-Producing Microorganisms in The
    Isolation and Characterization of Cellulase-Producing Microorganisms in the Red Sea Dissertation by Siham Kamal Fatani In Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy King Abdullah University of Science and Technology, Thuwal, Kingdom of Saudi Arabia September, 2019 2 Examination Committee Page The Dissertation of Siham Fatani is Examined by the Committee Members Committee Chairperson: Prof. Takashi Gojobori Committee Members: Prof. Vladimir Bajic, Prof. Susana Agusti, Prof. Shugo Watabe 3 © September, 2019 Siham Kamal Fatani All Rights Reserved 4 ACKNOWLEDGMENTS This work is a consequence of great help and guidance from many people; faculty, family and friends. I am really happy to have these people by my side while undertaking my PhD Dissertation. First, I would like to express my profound gratitude and respect to my supervisor, Prof. Takashi Gojobori, Distinguished Professor of Bioscience and Associate Director of Computational Bioscience Research Center for his professional guidance, and regular encouragement and motivation at various stages of this work. I would also like to thank Dr. Katsuhiko Mineta for his support and advice during my research. Moreover, I would like to express my deepest appreciation to Dr. Yoshimoto Saito for his assistance and suggestions throughout my project. I also appreciate Mr. Mohammad Al-Arawi for his support and technical advice and without them this work would not have been possible for me to complete. In addition, I would like to thank my committee members, Prof. Vladimir Bajic, Prof. Susana Agusti and Prof. Shugo Watabe for giving their time to review my Ph.D. thesis and for offering their insight and suggestions.
    [Show full text]
  • Long-Term Follow-Up and Sudden Unexpected Death in Gaucher Disease Type 3 in Egypt
    Long-term follow-up and sudden unexpected death in Gaucher disease type 3 in Egypt Magy Abdelwahab, MD, ABSTRACT PhD Objective: To describe the long-term follow-up and distinct phenotype of a large cohort of patients Derek Blankenship, PhD with Gaucher disease type 3 on enzyme replacement therapy (ERT) in Egypt. Raphael Schiffmann, Methods: A prospective cohort study of 78 patients on ERT who were followed for up to 9 years MD, MHSc with yearly evaluations that included EEG and cognitive testing. Results: Of the patients, 73% were homozygous for the L444P GBA1 mutation; all but 7 were Correspondence to neurologically symptomatic. Supranuclear gaze palsy with variable but stable cognitive function Dr. Schiffmann: was present in 91% of patients. Convergent strabismus and bulbar dysfunction were noted in Raphael.schiffmann@ baylorhealth.edu or 22% and 37%, respectively. Features of oppositional defiant disorder were present in 54% of Dr. Abdelwahab: patients. Twenty-three patients (30%) developed seizures while on ERT for 1–9 years. Of those, [email protected] 12 patients (15%) died suddenly and unexpectedly at a mean age of 6.7 6 5.0 years (range 1.5– 18). Sudden death was usually associated with a seizure disorder or a terminal seizure, but 7 of 12 patients had a preceding normal EEG. An additional 11% had background slowing or epilep- togenic activity on EEG without clinical seizures. There were 3 familial cases of sudden unex- pected death. Conclusions: Despite having the most common GBA1 genotype known to be associated with neuronopathic Gaucher disease, patients with Gaucher disease type 3 in Egypt have a phenotype and a clinical outcome on ERT that are very different from those observed in other populations.
    [Show full text]
  • Intravenous Enzyme Replacement Therapy (ERT) for Gaucher Disease
    UnitedHealthcare® Oxford Clinical Policy Intravenous Enzyme Replacement Therapy (ERT) for Gaucher Disease Policy Number: PHARMACY 270.16 T2 Effective Date: April 1, 2021 Instructions for Use Table of Contents Page Related Policies Coverage Rationale ....................................................................... 1 • Acquired Rare Disease Drug Therapy Exception Prior Authorization Requirements ................................................ 3 Process Applicable Codes .......................................................................... 3 • Drug Coverage Guidelines Background ................................................................................... 3 • Experimental/Investigational Treatment Benefit Considerations .................................................................. 4 • Experimental/Investigational Treatment for NJ Plans Clinical Evidence ........................................................................... 4 U.S. Food and Drug Administration ............................................. 6 • Provider Administered Drugs – Site of Care References ..................................................................................... 7 Policy History/Revision Information ............................................. 8 Instructions for Use ....................................................................... 8 Coverage Rationale See Benefit Considerations This policy refers to the following drug products, all of which are intravenous enzyme replacement therapies used in the treatment of Gaucher disease:
    [Show full text]