DOCSLIB.ORG

Type 3 Lodothyronine Deiodinase: Cloning, in Vitro Expression, and Functional Analysis of the Placental Selenoenzyme

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Type 3 Lodothyronine Deiodinase: Cloning, in Vitro Expression, and Functional Analysis of the Placental Selenoenzyme Type 3 lodothyronine deiodinase: cloning, in vitro expression, and functional analysis of the placental selenoenzyme. D Salvatore, … , D L St Germain, P R Larsen J Clin Invest. 1995;96(5):2421-2430. https://doi.org/10.1172/JCI118299. Research Article Type 3 iodothyronine deiodinase (D3) catalyzes the conversion of T4 and T3 to inactive metabolites. It is highly expressed in placenta and thus can regulate circulating fetal thyroid hormone concentrations throughout gestation. We have cloned and expressed a 2.1-kb human placental D3 cDNA which encodes a 32-kD protein with a Km of 1.2 nM for 5 deiodination of T3 and 340 nM for 5' deiodination of reverse T3. The reaction requires DTT and is not inhibited by 6n- propylthiouracil. We quantitated transiently expressed D3 by specifically labeling the protein with bromoacetyl [125I]T3. The Kcat/Km ratio for 5 deiodination of T3 was over 1,000-fold that for 5' deiodination of reverse T3. Human D3 is a selenoenzyme as evidenced by (a) the presence of an in frame UGA codon at position 144, (b) the synthesis of a 32-kD 75Se-labeled protein in D3 cDNA transfected cells, and (c) the presence of a selenocysteine insertion sequence element in the 3' untranslated region of the mRNA which is required for its expression. The D3 selenocysteine insertion sequence element is more potent than that in the type 1 deiodinase or glutathione peroxidase gene, suggesting a high priority for selenocysteine incorporation into this enzyme. The conservation of this enzyme from Xenopus laevis tadpoles to humans implies an essential role for regulation of thyroid hormone inactivation during embryological development. […] Find the latest version: https://jci.me/118299/pdf Type 3 lodothyronine Deiodinase: Cloning, In Vitro Expression, and Functional Analysis of the Placental Selenoenzyme Domenico Salvatore,* Susan C. Low,* Maria Berry,* Ana Luiza Maia,* John W. Hamey,* Walburga Croteau,$ Donald L. St. Germain,$ and P. Reed Larsen* *Thyroid Division, Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston, Massachusetts 02115; and tDepartments of Medicine and Physiology, Dartmouth Medical School, Lebanon, New Hampshire 03756 Abstract of thyroxine (T4) and thyroid follicles in the fetal thyroid by 10-12 wk of gestation as well as the potential availability of Type 3 iodothyronine deiodinase (D3) catalyzes the conver- maternal thyroid hormone, the free concentration of the active sion of T4 and T3 to inactive metabolites. It is highly ex- thyroid hormone, 3,5,3 '-triiodothyronine (T3), is less than half pressed in placenta and thus can regulate circulating fetal that of maternal levels up to the time of delivery (1-3). The thyroid hormone concentrations throughout gestation. We physiological rationale for this circumstance is not well under- have cloned and expressed a 2.1-kb human placental D3 stood but is thought to permit the precise timing and regulation cDNA which encodes a 32-kD protein with a Km of 1.2 nM of T3 delivery to the CNS by coordination of the expression for 5 deiodination of T3 and 340 nM for 5' deiodination of and action of the type 25 '-iodothyronine deiodinase (for review reverse T3. The reaction requires DTT and is not inhibited see reference 4). This enzyme uses tissue T4 as a substrate to by 6n-propylthiouracil. We quantitated transiently ex- produce T3 locally and is the primary source of T3 for this organ pressed D3 by specifically labeling the protein with bro- (5). It is also possible that normal circulating T3 concentrations moacetyl [12"'I T3. The KatIK. ratio for 5 deiodination of could have deleterious effects on immature tissues or could T3 was over 1,000-fold that for 5' deiodination of reverse enhance the metabolic requirements of the fetus. T3. Human D3 is a selenoenzyme as evidenced by (a) the There are two principal mechanisms by which the circulat- presence of an in frame UGA codon at position 144, (b) ing fetal T3 concentration is maintained at low levels. One is the synthesis of a 32-kD 75Se-labeled protein in D3 cDNA that the type 1 iodothyronine deiodinase (Dl) in fetal liver is transfected cells, and (c) the presence of a selenocysteine expressed at low levels relative to those in adult life (for review insertion sequence element in the 3' untranslated region of see references 6 and 7). This reduces the extrathyroidal T3 the mRNA which is required for its expression. The D3 supply from this source. The second important factor in main- selenocysteine insertion sequence element is more potent taining low serum T3 concentrations is the expression of high than that in the type 1 deiodinase or glutathione peroxidase levels of the type 3 deiodinase (D3) in placenta of all species gene, suggesting a high priority for selenocysteine incorpo- so far examined (for review see references 4, 6-8). This deio- ration into this enzyme. The conservation of this enzyme dinase catalyzes the inner ring deiodination of T3 and T4 inacti- from Xenopus laevis tadpoles to humans implies an essential vating circulating iodothyronines as well as minimizing trans- role for regulation of thyroid hormone inactivation during placental passage of maternal hormone. D3 activity is also ex- embryological development. (J. Clin. Invest. 1995. 96:2421- pressed in brain, especially in the rat fetus, in fetal rat skin and 2430.) Key words: iodide peroxidase * thyroxine * thyroid intestine, as well as in embryonic chick liver (6, 7). In humans, hormones * selenium * selenocysteine placental D3 is sufficiently potent that instillation of 700 ,ug of T4 into amniotic fluid at term causes insignificant increases in Introduction the neonatal serum T3 concentration assessed 24 h later (9). The cDNAs encoding the DI enzymes of several species Thyroid hormone is critical to the normal of development the have been cloned, and all have been shown to contain the rare human central nervous system (CNS).' Despite the presence amino acid selenocysteine (10- 12). The requirement for sele- nium in the active center of this enzyme for maximum activity can explain the increased ratio of T4 to T3 in the Address correspondence to P. Reed Larsen, Thyroid Division, Brig- circulation ham and Women's Hospital and Harvard Medical School, Boston, and the significant decrease in DI activity in the liver and kidney MA 02115. Phone: 617-732-6762; FAX: 617-731-4718; E-mail: of rats made selenium deficient (13, 14). However, selenium [email protected] deficiency has no effect on placental D3 activity nor can a Receivedfor publication 26 June 1995 and accepted in revisedform selenium-labeled protein be identified in rat placental micro- 7 August 1995. somes ( 15, 16). This has led to the conclusion that this enzyme does not contain selenocysteine. On the other hand, St. Germain 1. Abbreviations used in this paper: BrAc, bromoacetyl; CNS, central et al. recently identified a T3-responsive cDNA found in Xeno- nervous system; Dl, type 1 iodothyronine deiodinase; D3, type 3 idodo- pus laevis tadpoles as one encoding a D3 enzyme (XD3) (17). thyronine deiodinase; GH, growth hormone; GTG, gold thioglucose; h, The XD3 protein is 50% identical to rat DI and contains an human; HEK, human embryonic kidney; nt, nucleotide; PE, phosphate/ in frame UGA codon and selenocysteine insertion EDTA buffer; sequence PTU, 6n-propylthiouracil; SECIS, selenocysteine inser- (SECIS) element in the 3'-untranslated of the tion sequence; SEl D, selenophosphate synthetase, TK, thymidine ki- region mRNA nase; ut, untranslated. (17). A SECIS element is a stem loop sequence which is re- quired for suppression of the stop codon function of UGA and J. Clin. Invest. the insertion of selenocysteine (18). We undertook the present © The American Society for Clinical Investigation, Inc. studies to identify the human D3, to establish whether it is, or 0021-9738/95/11/2421/10 $2.00 is not, a selenoenzyme and analyze its tissue expression and Volume 96, November 1995, 2421-2430 catalytic function. Human Type 3 Deiodinase 2421 Methods Preparation of hD3, XD3, and SelD vectors for eukaryotic expres- sion. To confirm that this clone encoded the type 3 deiodinase, a 1.9- kb insert containing 35 bp of 5' untranslated (ut) region (186-2066) Materials. Two cDNA libraries were used. The first was a human pla- (Fig 1) was excised from pBluescript and inserted into the XhoI/NotI centa cDNA library in a CDM-8 vector prepared according to the meth- sites of CDM-8, a mammalian expression vector (20). The same con- ods of Aruffo and Seed (20), and kindly provided by Dr. Brian Seed struct was then prepared with the Xenopus D3 cDNA (XD3), kindly (Massachusetts General Hospital, Boston, MA). The second, a human provided by Drs. Robert A. Schwartzman and Donald D. Brown of placenta cDNA library in X-Zap II, was purchased from Stratagene Inc. Carnegie Institution (Baltimore, MD), that was excised from pBS with (La Jolla, CA). Bromoacetylchloride was from Aldrich Chemical Co. EcoRI/XhoI restriction enzymes and subcloned into the same site of (Milwaukee, WI). '25I-labeled 3,3',5'-triiodothyronine (rT3) and ['25I] the D10 eukaryotic expression vector (21). 3,5,3' triiodothyronine were from DuPont-NEN (Boston, MA). [75Se] - The human selenophosphate synthetase (Sel D), the homologue of Selenite was a generous gift of Dr. Dolph Hatfield (National Institutes the bacterial enzyme (22), has recently been cloned (23). Its cDNA of Health, Bethesda, MD) and was originally obtained from Dr. Kurt was also subcloned into CDM-8 as described. Zinn (University of Missouri Research Reactor, Columbia, MO). All DNA transfections. Transfection of human embryonic kidney other chemicals were of reagent grade. (HEK)-293 cells was by calcium phosphate precipitation as described Isolation of a human D3 cDNA. Based on sequence homology be- previously (24).
Load more

Recommended publications
  • Illness-Induced Changes in Thyroid Hormone Metabolism: Focus on the Tissue Level
    r e V i e W illness-induced changes in thyroid hormone metabolism: focus on the tissue level J. Kwakkel*, E. Fliers, A. Boelen Department of Endocrinology & Metabolism, Academic Medical Center, University of Amsterdam, the Netherlands, *corresponding author: tel.: +31 (0)20-566 67 01, fax: +31 (0)20-691 76 82, e-mail: [email protected] a b s t r a C t during illness changes in thyroid hormone metabolism ring and the outer (tyrosyl) ring of T4 can be deiodinated, occur, collectively known as the non-thyroidal illness ultimately leading to the formation of 3,3’-di-iodothyronine syndrome (NTIS). NTIS is characterised by low serum (T2) (figure 1). thyroid hormone levels without the expected rise in serum thyroid-stimulating hormone, indicating a major change in thyroid hormone feedback regulation. recent studies n o n - t H yroidal illness syndro M e have made clear that during NTIS differential changes in thyroid hormone metabolism occur in various tissues, the During illness many aspects of thyroid hormone net effect of which may be either activation or inhibition of metabolism change, collectively known as the thyroid hormone action. in this review we discuss systemic non-thyroidal illness syndrome (NTIS). The hallmark of and local changes in thyroid hormone metabolism during NTIS is decreased serum thyroid hormone levels without illness, highlighting their physiological implications in an increase in TSH and TRH expression, indicating terms of disease course. the absence of negative feedback regulation. This may represent a useful adaptation of the body to counteract excessive catabolism observed during illness and can be K e y W o r d s viewed as a part of the acute phase response.4 However, especially during prolonged critical illness in the ICU Deiodinase, inflammation, non-thyroidal illness syndrome, setting NTIS may be maladaptative.5 thyroid hormone figure 1.
    [Show full text]
  • Thyroxine Binding to Type III Iodothyronine Deiodinase Craig A
    www.nature.com/scientificreports OPEN Thyroxine binding to type III iodothyronine deiodinase Craig A. Bayse*, Eric S. Marsan, Jenna R. Garcia & Alexis T. Tran‑Thompson Iodothyronine deiodinases (Dios) are important selenoproteins that control the concentration of the active thyroid hormone (TH) triiodothyronine through regioselective deiodination. The X‑ray structure of a truncated monomer of Type III Dio (Dio3), which deiodinates TH inner rings through a selenocysteine (Sec) residue, revealed a thioredoxin-fold catalytic domain supplemented with an unstructured Ω-loop. Loop dynamics are driven by interactions of the conserved Trp207 with solvent in multi-microsecond molecular dynamics simulations of the Dio3 thioredoxin(Trx)-fold domain. Hydrogen bonding interactions of Glu200 with residues conserved across the Dio family anchor the loop’s n‑terminus to the active site Ser‑cys-Thr‑Sec sequence. A key long‑lived loop conformation coincides with the opening of a cryptic pocket that accommodates thyroxine (T4) through an I⋯Se halogen bond to Sec170 and the amino acid group with a polar cleft. The Dio3-T4 complex is stabilized by an I⋯O halogen bond between an outer ring iodine and Asp211, consistent with Dio3 selectivity for inner ring deiodination. Non-conservation of residues, such as Asp211, in other Dio types in the fexible portion of the loop sequence suggests a mechanism for regioselectivity through Dio type- specifc loop conformations. Cys168 is proposed to attack the selenenyl iodide intermediate to regenerate Dio3 based upon structural comparison with related Trx-fold proteins. Iodothyronine deiodinase (Dio) membrane selenoproteins regulate thyroid hormone (TH) activity through regioselective deiodination (Fig. 1a)1–10.
    [Show full text]
  • The Interactions Between Selenium and Iodine Deficiencies in Man And
    Nutrition Research Reviews (1999), 12, 55±73 55 The interactions between selenium and iodine de®ciencies in man and animals John R. Arthur1*, Geoffrey J. Beckett2 and Julie H. Mitchell1 1Division of Micronutrient and Lipid Metabolism, Rowett Research Institute, Greenburn Road, Bucksburn, Aberdeen AB21 9SB, UK 2University Department of Clinical Biochemistry, The Royal In®rmary, Edinburgh EH3 9YW, UK Abstract Up to one billion people live in areas where they may be at risk from I de®ciency. Many of the debilitating effects of the de®ciency may be irreversible, consequently it is essential to understand the mechanisms whereby lack of I can cause disease 0 through decreased thyroxine and 3,3 ,5-triiodothyronine (T3) synthesis. Since Se has an essential role in thyroid hormone metabolism, it has the potential to play a major part in the outcome of I de®ciency. These effects of Se derive from two aspects of its biological function. First, three Se-containing deiodinases regulate the synthesis and degradation of the biologically active thyroid hormone, T3. Second, selenoperoxidases and possibly thioredoxin reductase (EC 1.6.4.5) protect the thyroid gland from H2O2 produced during the synthesis of thyroid hormones. The mechanisms whereby Se de®ciency exacerbates the hypothyroidism due to I de®ciency have been elucidated in animals. In contrast to these adverse effects, concurrent Se de®ciency may also cause changes in deiodinase activities which can protect the brain from low T3 concentrations in I de®ciency. Animals with Se and I de®ciency have changes in serum thyroid hormone concentrations that are similar to those observed in patients with I de®ciency disease.
    [Show full text]
  • 74Th Annual Meeting Program
    74th Annual Meeting of the American Thyroid Association Millennium Biltmore Hotel Los Angeles, California October 10 – 13, 2002 Thursday, October 10, 2002 6:00 – 7:45 am Crystal Ballroom Defining Thyroid Hypofunction: Perspectives from Populations and the Clinic Moderator: Paul W. Ladenson The Spectrum of Thyroid Hypofunction in Populations E. Chester Ridgway Getting Practical: How to Treat Patients with Thyroid Hypofunction Gilbert H. Daniels Case Discussion: Now It’s Normal, Now It’s Not Paul W. Ladenson “Early Riser” CME Symposium and breakfast supported by an unrestricted educational grant from Abbott Laboratories 8:00 – 8:15 am Biltmore Bowl Welcome and Introductions Carole A. Spencer ATA President Paul W. Ladenson ATA Secretary Gregory A. Brent ATA Program Chair, Scientific Leonard Wartofsky ATA Program Chair, Clinical 8:15 – 9:00 am Biltmore Bowl Keynote Clinical Address Thyroid Disease in Pregnancy Daniel Glinoer, MD, PhD University Hospital St. Pierre Department of Internal Medicine Brussels, Belgium Supported by an educational grant from Abbott Laboratories 9:00 – 10:00 am Biltmore Bowl Plenary Session – Topic Highlights Oral Abstract Presentations Chairs: Gregory A. Brent and Leonard Wartofsky 1 9:00 am Thyroid Hormone Metabolism The Gene Coding for the Type 3 Iodothyronine Deiodinase Is Imprinted and Required for Normal Neonatal Growth and Survival A. Hernandez1,2, S. Fiering3, E. Martinez1,2, V. Galton2, D. St. Germain1,2 Departments of 1Medicine, 2Physiology, and 3Microbiology and Immunology, Dartmouth Medical School, Lebanon, New Hampshire, USA 57 Thursday, October 10, 2002 Morning Session 2 9:15 am Thyroid Diseases NHANES III: Impact of TSH:TPOAb Relationships on Redefining the Serum TSH Normal Reference Range C.
    [Show full text]
  • Selenium-Containing Enzymes in Mammals: Chemical Perspectives
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Publications of the IAS Fellows J. Chem. Sci., Vol. 117, No. 4, July 2005, pp. 287–303. © Indian Academy of Sciences. Selenium-containing enzymes in mammals: Chemical perspectives GOURIPRASANNA ROY, BANI KANTA SARMA, PRASAD P PHADNIS and G MUGESH* Department of Inorganic and Physical Chemistry, Indian Institute of Science, Bangalore 560 012, India e-mail: [email protected] MS received 22 March 2005; accepted 6 June 2005 Abstract. The chemical and biochemical route to the synthesis of the 21st amino acid in living systems, selenocysteine, is described. The incorporation of this rare amino acid residue into proteins is described with emphasis on the role of monoselenophosphate as selenium source. The role of selenocysteine moiety in natural mammalian enzymes such as glutathione peroxidase (GPx), iodothyronine deiodinase (ID) and thioredoxin reductase (TrxR) is highlighted and the effect of other amino acid residues located in close proximity to selenocysteine is described. It is evident from various studies that two amino acid residues, tryptophan and glutamine, appear in identical positions in all known members of the GPx family. Ac- cording to the three-dimensional structure established for bovine GPx, these residues could constitute a catalytic triad in which the selenol group of the selenocysteine is both stabilized and activated by hydro- gen bonding with the imino group of the tryptophan (Trp) residue and with the amido group of the gluta- mine (Gln) residue. The ID enzymes, on the other hand, do not possess any Trp or Gln residues in close proximity to selenium, but contain several histidine residues, which may play important roles in the ca- talysis.
    [Show full text]
  • Characterization of Cytosolic Glutathione Peroxidase And
    Aquatic Toxicology 130–131 (2013) 97–111 Contents lists available at SciVerse ScienceDirect Aquatic Toxicology jou rnal homepage: www.elsevier.com/locate/aquatox Characterization of cytosolic glutathione peroxidase and phospholipid-hydroperoxide glutathione peroxidase genes in rainbow trout (Oncorhynchus mykiss) and their modulation by in vitro selenium exposure a a b a d c a,∗ D. Pacitti , T. Wang , M.M. Page , S.A.M. Martin , J. Sweetman , J. Feldmann , C.J. Secombes a Scottish Fish Immunology Research Centre, Institute of Biological and Environmental Sciences, University of Aberdeen, Aberdeen AB24 2TZ, United Kingdom b Integrative and Environmental Physiology, Institute of Biological and Environmental Sciences, University of Aberdeen, Aberdeen AB24 2TZ, United Kingdom c Trace Element Speciation Laboratory, Department of Chemistry, University of Aberdeen, Aberdeen AB24 3UE, United Kingdom d Alltech Biosciences Centre, Sarney, Summerhill Rd, Dunboyne, Country Meath, Ireland a r t i c l e i n f o a b s t r a c t Article history: Selenium (Se) is an oligonutrient with both essential biological functions and recognized harmful effects. Received 4 July 2012 As the selenocysteine (SeCys) amino acid, selenium is integrated in several Se-containing proteins Received in revised form (selenoproteins), many of which are fundamental for cell homeostasis. Nevertheless, selenium may exert 19 December 2012 toxic effects at levels marginally above those required, mainly through the generation of reactive oxygen Accepted 20 December 2012 species (ROS). The selenium chemical speciation can strongly affect the bioavailability of this metal and its impact on metabolism, dictating the levels that can be beneficial or detrimental towards an organism.
    [Show full text]
  • In Vitro & Ex Vivo Assays for Identification of Modulators
    DRAFT IN VITRO & EX VIVO ASSAYS FOR IDENTIFICATION OF MODULATORS OF THYROID HORMONE SIGNALLING PART TWO: COMPENDIUM OF IN VITRO & EX VIVO ASSAYS FOR THYROID HORMONE DISRUPTORS 11/10/213 BLOCK #1 ASSAYS 3 1.1 Hypothalamic Thyrotropin-releasing hormone (TRH) production/release assays 3 1.2 Thyrotropin Releasing Hormone Receptor Activation Assays 4 1.3 Thyrotropin Receptor Activation Assays 5 BLOCK #2 ASSAYS 6 2.1 Thyroperoxidase Inhibition Assay 6 2.2. Sodium/Iodide Symporter Mediated Uptake Assays 10 BLOCK #3 ASSAYS 14 3.1. Combined TBG and Transthyretin TTR Binding Assays 14 3.1a TransThyretin (TTR) Binding Assays 17 BLOCK #4 ASSAYS 19 4.1 Deiodinase Activity Assays 19 4.2 Glucuronidation Assays 22 4.3 Sulfation Assays 24 BLOCK #5 ASSAYS 25 5.1 Thyroid Transmembrane Transporter Assays 25 BLOCK #6 ASSAYS 31 6.1 TR Binding Assays 31 6.2 TR Transactivation Assys 37 6.3. Assays that evaluate interactions of TR with other regulatory proteins 38 1 BLOCK #7 ASSAYS 39 7.1. Zebrafish Assays 39 BLOCK #8 ASSAYS 40 8.1. T-screen Assays 40 8.2. Developmental Neurotoxicity in vitro Assay 41 REFERENCES 42 2 BLOCK #1 ASSAYS CENTRAL REGULATION: HYPOTHALAMIC-PITUITARY-THYROID AXIS (HPT) This block includes assays with a potential to evaluate synthesis/production of Thyrotropin Releasing Hormone (TRH) at the hypothalamus level, the function/activation of the TRH-Receptor at the pituitary level and of the Thyroid Stimulating Hormone (TSH) Receptor (at the thyroid level). 1.1 Hypothalamic Thyrotropin-releasing hormone (TRH) production/release assays Assay Name Hypothalamic TRH production and release Molecular Low T3 levels, increased leptin/ MC4R signalling, modulated cAMP levels in the TRH Initiation Event or producing neurons in the paraventricular nucleus of the hypothalamus Key Event Endpoint(s)/ purpose of the Assess effects on chemicals on T3 repression on TRH transcription and production.
    [Show full text]
  • (Mpo421-8B2) Nbp2-41406
    Product Datasheet Myeloperoxidase/MPO Antibody (MPO421-8B2) NBP2-41406 Unit Size: 0.1 mg Store at 4C. Do not freeze. Publications: 1 Protocols, Publications, Related Products, Reviews, Research Tools and Images at: www.novusbio.com/NBP2-41406 Updated 12/1/2020 v.20.1 Earn rewards for product reviews and publications. Submit a publication at www.novusbio.com/publications Submit a review at www.novusbio.com/reviews/destination/NBP2-41406 Page 1 of 3 v.20.1 Updated 12/1/2020 NBP2-41406 Myeloperoxidase/MPO Antibody (MPO421-8B2) Product Information Unit Size 0.1 mg Concentration 1 mg/ml Storage Store at 4C. Do not freeze. Clonality Monoclonal Clone MPO421-8B2 Preservative 15mM Sodium Azide Isotype IgG1 Purity Protein A purified Buffer PBS (pH 7.4) Product Description Host Mouse Gene ID 4353 Gene Symbol MPO Species Human Specificity/Sensitivity The mouse monoclonal antibody MPO421-8B2 recognizes human myeloperoxidase, a heme protein present in myeloblasts, neutrophils and monocytes. It is a marker of acute myelogenous leukemias and acute lymphoblastic leukemias. Immunogen Human myeloperoxidase Product Application Details Applications ELISA, Flow Cytometry, Flow (Intracellular), CyTOF-ready Recommended Dilutions Flow Cytometry 1-4 ug/ml, ELISA, Flow (Intracellular) 1 ug/ml, CyTOF-ready Application Notes Use in FLOW cytometry reported in scientific literature ( PMID 27355490). This antibody is CyTOF ready. Page 2 of 3 v.20.1 Updated 12/1/2020 Images Flow (Intracellular): Myeloperoxidase/MPO Antibody (MPO421-8B2) [NBP2-41406] - Separation of human neutrophil granulocytes (red-filled) from lymphocytes (black-dashed) in flow cytometry analysis (intracellular staining) of human peripheral whole blood stained using anti-human Myeloperoxidase (MPO421-8B2) purified antibody (concentration in sample 1 ug/ml).
    [Show full text]
  • Thyroid Screen (Serum)
    Thyroid Screen (Serum) Patient: Order Number: DOB: Completed: Sex: F Received: MRN: Collected: Sample Type - Serum Result Reference Range Units Central Thyroid Regulation & Activity Total Thyroxine (T4) 75 75 58-161 nmol/L Thyroid Stimulating 3.97 3.97 0.40-4.00 microIU/mL Hormone (TSH) Free Thyroxine (FT4) 12.9 12.9 11.5-22.7 pmol/L Peripheral Thyroid Function Free T3 4.4 4.4 2.8-6.5 pmol/L FT4 : FT3 Ratio 2.9 Thyroid Auto Immunity Thyroglobulin (TG) 43 H 43 <= 40 IU/mL Peroxidase (TPO) 143 H 143 <= 34 IU/mL Key Guide Testing performed by Genova Diagnostics, Inc. 63 Zillicoa St., Asheville, NC 28801-0174 Inside Reference Range Outside Reference Range © Genova Diagnostics · A. L. Peace-Brewer, PhD, D(ABMLI), Lab Director · CLIA Lic. #34D0655571 · Medicare Lic. #34-8475 BTSCN Page 2 Patient: ID: Thyroid Metabolism at a Glance Pituitary TSH 3.97 Auto Immunity Central Anti - - TPO Antibody Regulation Thyroid Gland Free T4 143 12.9 Anti - - TG Antibody small amounts of T3 43 5' deiodinase 5 deiodinase Se dependent Peripheral Regulation Free T3 Reverse T3 inactive 4.4 most active 5' deiodinase 5 deiodinase Se dependent T₂ Inactive T₂ Inactive © Genova Diagnostics · A. L. Peace-Brewer, PhD, D(ABMLI), Lab Director · CLIA Lic. #34D0655571 · Medicare Lic. #34-8475 Patient: ID: Page 3 Commentary Please note the reference range for Peroxidase (TPO) has been updated due to a methodology update. Commentary is provided to the practitioner for educational purposes, and should not be interpreted as diagnostic or treatment recommendations. Diagnosis and treatment decisions are the responsibility of the practitioner.
    [Show full text]
  • Assessment of Type 1 and Type 3 Deiodinase Expression Levels In
    Research paper Acta Neurobiol Exp 2017, 77: 225–235 Assessment of type 1 and type 3 deiodinase expression levels in depressive disorders Elżbieta Gałecka1*, Anna Kumor‑Kisielewska1, Agata Orzechowska2, Michael Maes3,4,5, Paweł Górski1, and Janusz Szemraj6 1 Department of Pulmonology and Allergology, Medical University of Lodz, Poland, 2 Department of Adult Psychiatry, Medical University of Lodz, Poland, 3 Deakin University IMPACT Strategic Research Centre, Deakin University, School of Medicine, Barwon Health, Geelong, Australia, 4 Department of Psychiatry, Chulalongkorn University, Bangkok, Thailand, 5 Health Sciences Graduate Program, Health Sciences Center, State University of Londrina, Brazil, 6 Department of Medical Biochemistry, Medical University of Lodz, Poland, * Email: [email protected] A depressive disorder is a disease characterized by a heterogenous background. The important processes observed and diagnosed in depressed patients indicate that the etiology of depression may include disturbances in thyroid hormone (TH) levels and the occurrence of immune‑inflammatory activation. Type 1 (DIO1) and type 3 (DIO3) iodothyronine deiodinases are the enzymes which determine the peripheral and tissue levels of TH, but also interfere with immunological cells and inflammatory processes. We aimed to investigate the levels of DIO1 and DIO3 in the patients suffering from recurrent depressive disorders (rDD). Data collected from 91 rDD patients and 105 healthy controls were analyzed. The diagnoses were made based on the ICD‑10 criteria (F33.0–F33.8). The expression levels of DIO1 and DIO3 were estimated using the polymerase chain reaction method and the enzyme‑linked immunosorbent assay (ELISA). The expression of DIO1 on mRNA/protein levels in the rDD patients was reduced in comparison to the control subjects, while the expression of DIO3 was higher in the patients suffering from depression.
    [Show full text]
  • Evidence That a Polymorphism Within the 3 UTR of Glutathione
    Genes Nutr (2007) 2:225–232 DOI 10.1007/s12263-007-0052-3 RESEARCH PAPER Evidence that a polymorphism within the 30UTR of glutathione peroxidase 4 is functional and is associated with susceptibility to colorectal cancer G. Bermano Æ V. Pagmantidis Æ N. Holloway Æ S. Kadri Æ N. A. G. Mowat Æ R. S. Shiel Æ J. R. Arthur Æ J. C. Mathers Æ A. K. Daly Æ J. Broom Æ J. E. Hesketh Received: 24 November 2006 / Accepted: 18 January 2007 / Published online: 13 October 2007 Ó Springer-Verlag 2007 Abstract Low selenium (Se) status has been associated study was carried out in cohorts of patients with either with increased risk of colorectal cancer (CRC). Se is adenomatous polyps, colorectal adenocarcinomas and in present as the amino acid selenocysteine in selenoproteins, healthy controls. A higher proportion of individuals with such as the glutathione peroxidases. Se incorporation CC genotype at the GPx4 T/C 718 SNP was present in the requires specific RNA structures in the 30 untranslated cancer group, but not in the polyp group, compared with region (30UTR) of the selenoprotein mRNAs. A single the control group (P \ 0.05). The present data demonstrate nucleotide polymorphism (SNP) occurs at nucleotide 718 the functionality of the GPx4 T/C 718 SNP and suggest (within the 30UTR) in the glutathione peroxidase 4 gene. In that T genotype is associated with lower risk of CRC. the present study, Caco-2 cells were transfected with constructs in which type 1 iodothyronine deiodinase coding Keywords Colorectal cancer Á GPx4 Á Reporter gene Á region was linked to the GPx4 30UTR with either C or T Selenium Á SNP Á 30Untranslated region variant at position 718.
    [Show full text]
  • Thermogenesis in Adipose Tissue Activated by Thyroid Hormone
    International Journal of Molecular Sciences Review Thermogenesis in Adipose Tissue Activated by Thyroid Hormone Winifred W. Yau 1 and Paul M. Yen 1,2,* 1 Laboratory of Hormonal Regulation, Cardiovascular and Metabolic Disorders Program, Duke NUS Medical School, Singapore 169857, Singapore; [email protected] 2 Duke Molecular Physiology Institute, Duke University, Durham, NC 27708, USA * Correspondence: [email protected]; Tel.: +65-6516-7666 Received: 23 March 2020; Accepted: 22 April 2020; Published: 24 April 2020 Abstract: Thermogenesis is the production of heat that occurs in all warm-blooded animals. During cold exposure, there is obligatory thermogenesis derived from body metabolism as well as adaptive thermogenesis through shivering and non-shivering mechanisms. The latter mainly occurs in brown adipose tissue (BAT) and muscle; however, white adipose tissue (WAT) also can undergo browning via adrenergic stimulation to acquire thermogenic potential. Thyroid hormone (TH) also exerts profound effects on thermoregulation, as decreased body temperature and increased body temperature occur during hypothyroidism and hyperthyroidism, respectively. We have termed the TH-mediated thermogenesis under thermoneutral conditions “activated” thermogenesis. TH acts on the brown and/or white adipose tissues to induce uncoupled respiration through the induction of the uncoupling protein (Ucp1) to generate heat. TH acts centrally to activate the BAT and browning through the sympathetic nervous system. However, recent studies also show that TH acts peripherally on the BAT to directly stimulate Ucp1 expression and thermogenesis through an autophagy-dependent mechanism. Additionally, THs can exert Ucp1-independent effects on thermogenesis, most likely through activation of exothermic metabolic pathways. This review summarizes thermogenic effects of THs on adipose tissues.
    [Show full text]