DOCSLIB.ORG

Formation of 10-Formylfolic Acid, a Potent Inhibitor of Dihydrofolate

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Formation of 10-Formylfolic Acid, a Potent Inhibitor of Dihydrofolate Proc. Nat. Acad. Sci. USA Vol. 71, No. 7, pp. 2736-2739, July 1974 Formation of 10-Formylfolic Acid, a Potent Inhibitor of Dihydrofolate Reductase, in Rat Liver Slices Incubated with Folic Acid (folate metabolism/folate coenzymes/natural folate enzyme inhibitors/regulation) M. d'URSO-SCOTT, J. UHOCH, AND J. R. BERTINO Yale University School of Medicine, Departments of Pharmacology and Medicine, New Haven, Connecticut 06510 Communicated by Aaron B. Lerner, April 15, 1974. ABSTRACT During investigations of folate poly- pared by reduction of lO-formyl FA (11) and subsequent acid glutamate biosynthesis in rat liver slices utilizing I2-14C]_ treatment (12). Materials were checked periodically for folic acid, a folate compound that behaved like a poly- glutamate form in the Sephadex G-15 gel filtration system purity by Sephadex G-15 and DEAE Sephadex A-25 column was found to accumulate. Subsequent chromatographic, chromatography (13). spectral, chemical, and enzymic studies have indicated that the compound formed in liver slices incubated with Rat Liver Slice Incubation. Adult male Sprague-Dawley [14Clfolic acid with and without methotrexate was 10- rats weighing approximately 300 g were killed and bled by formyl folate. This folate is of interest in that it is the decapitation. The livers were removed rapidly and chilled most potent natural inhibitor of dihydrofolate reductase on ice. Liver slices were prepared with a Stadie-Riggs hand known and may be capable of serving a regulatory func- tion within the cell. microtome. Incubations were performed using 1.0 g of liver slices in 10 ml of Krebs-Ringer bicarbonate buffer with 10-Formylpteroic acid (rhizopterin), a naturally occurring glucose (5.5 mM) containing 1 uCi of [14C]FA. In certain compound, was first isolated from cultures of Rhizopus nigri- experiments MTX (1 uMM) was included and in some experi- cans (1, 2). Its structure was confirmed by chemical synthesis, ments unlabeled carrier FA (0.1 mM) was present. The flasks degradation product analysis, and the preparation of model containing tissue slices and incubation mix were incubated at compounds (2, 3). Rhizopterin will support the growth of 370 with shaking for 4-6 hr under an atmosphere of 5% Streptococcus faecalis but not Lactobacillus casei or Pediococcus C02-95% 02. A typical incubation of liver slices gave ap- cerevisiae (4). One of the model compounds synthesized was proximately 4 nmole of 10-formylfolic acid/hr per g of tissue 10-formylfolic acid (10-formyl FA) formed by treatment of (wet weight). folic acid (FA) with concentrated formic acid (5). 10-formyl FA subsequently isolated from horse liver (6) was found to be Extraction of Radiolabeled Folates. At the end of the incuba- tion, liver slices were washed with ice-cold saline to remove active as a growth factor for S. faecalis and L. casei but not slices for P. cerevisiae (4). This substance has some unusual proper- any external label. Folates were extracted by dropping of liver tissue into one volume of boiling 1% ascorbate (pH ties for a folate compound, such as its fluorescence character- After 5 of the and its ultraviolet spectrum at neutral pH. In addition, 6.0) to minimize autolysis (14). min boiling, istics were and the an of FA is its potent in- slices homogenized and centrifuged supernatant unexpected property 10-formyl treated as described below. hibition of a key enzyme in folate metabolism, dihydrofolate solution was reductase (5.6,7,8-tetrahydrofolate:NADP+ oxidoreductase, Sephadex G-15 Gel Filtration. Samples were treated accord- EC 1.5.1.3) (7). ing to the procedure developed by Shin, et al. (15). A 2.4 X The present paper demonstrates that liver slices, when 45-cm glass column was packed with Sephadex G-15 gel incubated with [14C]FA, accumulate 10-formyl FA. The which had been previously equilibrated with eluent buffer. identity of this compound was not suspected initially because The elution was carried out with 0.02 M K2HPO4-KH2PO4 of its anomalous behavior on Sephadex G-15, namely, it buffer (pH 7.0) and the eluate collected in 5.5-ml fractions at behaved like a pteroyldiglutamate, rather than a pteroyl- 40 in darkness. In certain experiments, ascorbate (1%) or monoglutamate. 2-mercaptoethanol (0.2 M) was added to the buffer in order MATERIALS AND METHODS to protect labile folate forms. Miaterials. [2-14C]Folic acid (Q[4C]FA), specific activity DEAE Sephadex A-25 Chromatography. DEAE Sephadex 55.3 ,Ci/,umole, was purchased from Amersham/Searle. A-25 was prepared for anion exchange by washing with Methotrexate sodium salt (MTX) was purchased from Lederle 1 M NaOH, water, 1 M HCl, and water again and then equili- Laboratories and unlabeled folic acid was purchased from brated with 0.1 M NaCl. Samples were eluted with a linear Nutritional Biochemical Corp. Pteroyl-'y-glutamyl-glutamic gradient from 0.1 M NaCl to 0.7 M NaCl in 0.01 M K2HPO4- acid (pteroyldiglutamate), was synthesized by the method of KH2PO4buffer (pH 7.0), a modification of the procedure de- Baugh et al. (8, 9). 10-Formyl FA was prepared by formylation veloped by Baugh et al. (16). A 0.9 X 30-cm column was used of FA (10). 5,10-Methenyltetrahydrofolic acid was pre- and 2.7-ml fractions were collected at 40 in darkness. Abbreviations: FA, folic acid (= pteroylglutamic acid); MTX, Enzyme Assays. Carboxypeptidase G, was obtained from methotrexate sodium salt. Pseudomonas stutzeri and assayed as previously described 2736 Downloaded by guest on October 2, 2021 Proc. Nat. Acad. Sci. USA 71 (1974) Formation of lO-Formyl Folate 2737 E CL 500 - V0 0 20 40 60 80 100 FRACliON NO. FIG. 2. Sephadex gel filtration of liver slice extracts from a 0 20 40 60 80 100 ['4C]folate incubation mixture containing MTX. After 4 hr of FRACilON NO. incubation, in the presence of 1 ,uM MTX, liver slices were treated and the extract filtered through Sephadex G-15 as described in FIG. 1. Sephadex gel filtration of liver slice extracts. Liver the text. extract, prepared as described in the Materials and Methods section, was applied to a Sephadex G-15 column. The letters Since folate compounds formylated in the 10-position are indicate the elution positions of reference compounds: (a) known to have less affinity than other folate forms for anion pteroylpentaglutamate; (b) pteroyltriglutamate; (c) pteroyl- like DEAE-cellulose and since the folate in diglutamate; (d) 5-formyltetrahydrofolic acid; (e) dihydrofolic exchangers (13), acid, tetrahydrofolic acid, 5-methyltetrahydrofolic acid. (The question was presumably not a reduced form, there was a last four compounds are all pteroylmonoglutamate derivatives.) strong possibility that this compound was 10-formyl FA. Accordingly, authentic unlabeled 10-formyl FA was added to the liver slice extract as a marker and was found to cochro- (17). The material was homogeneous on polyacrylamide gel matograph both on Sephadex G-15 and on DEAE Sephadex electrophoresis, and had a specific activity of 700 units/mg of A-25 (Fig. 3) with the radioactive compound from the liver protein. To the radioactive folate formed in from liver slice extract. confirm that Pure dihydrofolate reductase was prepared rat liver slices was 10-formyl FA, the following experiments were by affinity chromatography on MTX-Sepharose (18). The specific activity of the material utilized was 50 units/mg of performed. protein as determined by the method of Perkins et al. (19). Purification of the Radioactive Folate From Liver Slices was ac- complished by incubating 100 g of rat liver slices with ['4C]FA Ultraviolet and Fluorescent Spectra. Identification of folates (10,uCi) and unlabeled FA (0.1 mM) in the presence of MTX and localization of chromatographic markers of CPG1 assays for 6 hr. The liver slices were washed, boiled in ascorbate, were monitored on a Cary model 15 spectrophotometer. homogenized, and centrifuged as described in Materials and Dihydrofolate reductase assays were carried out on a Gilford Methods. The supernatant solution was purified in batches 2000 spectrophotometer and fluorescence spectra were deter- by filtration through Sephadex G-15 followed by chroma- mined with an Aminco-Bowman spectrophotofluorometer. tography on DEAE Sephadex A-25. The appropriate fractions RESULTS were pooled, lyophilized, reconstituted in 0.1 M K2HPO4- KH2PO4 buffer (pH 7.0), and used for spectral determinations. Column Chromatography. When rat liver slices incubated with [14C]FA were extracted into ascorbate and the extract Ultraviolet and Fluorescence Spectra. Difference spectra filtered through Sephadex G-15 the major portion of radio- were taken because of the presence in the liver slice extract activity eluted early, in fractions 24 to 42 (Fig. 1). This por- of an impurity that absorbed maximally at 260 nm and inter- tion eluted well before any of the folate monoglutamate fered with the ultraviolet absorbance spectra. Comparison forms tested (MTX, FA, dihydrofolic acid, tetrahydrofolic of the difference spectra of authentic lO-formyl FA and the acid, 5-methyltetrahydrofolic acid, and 5-formyltetrahydro- radioactive folate formed in liver slices (Fig. 4) shows an folic acid) and was initially considered to be a folate poly- almost identical spectral change occurring in both samples glutamate, probably a di- or triglutamyl form. with the addition of NaOH to pH 13. In order to determine if the separated substances obtained Since lO-formyl FA fluoresces strongly when activated at were oxidized or reduced folates, the liver slices were incubated 364 nm with an emission X max at 450-460 nm, the partially with [14C]FA in the presence of MTX, a potent inhibitor of purified liver extract was tested for fluorescence. The material the enzymic reduction of FA or dihydrofolic acid (20, 21). was found to exhibit a strong 458-nm fluorescence maximum When the liver extract was filtered through Sephadex G-15 at neutral pH, behavior identical to that of lO-formyl FA.
Load more

Recommended publications
  • Methionine Synthase Supports Tumor Tetrahydrofolate Pools
    bioRxiv preprint doi: https://doi.org/10.1101/2020.09.05.284521; this version posted September 7, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Methionine synthase supports tumor tetrahydrofolate pools Joshua Z. Wang1,2,#, Jonathan M. Ghergurovich1,3,#, Lifeng Yang1,2, and Joshua D. Rabinowitz1,2,* 1 Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, New Jersey, USA 2 Department of Chemistry, Princeton University, Princeton, New Jersey, USA 3 Department of Molecular Biology, Princeton University, Princeton, New Jersey, USA # These authors contributed equally to this work. *Corresponding author: Joshua Rabinowitz Department of Chemistry and the Lewis-Sigler Institute for Integrative Genomics, Princeton University, Washington Rd, Princeton, NJ 08544, USA Phone: (609) 258-8985; e-mail: [email protected] Conflict of Interest Disclosure: J.D.R. is a paid advisor and stockholder in Kadmon Pharmaceuticals, L.E.A.F. Pharmaceuticals, and Rafael Pharmaceuticals; a paid consultant of Pfizer; a founder, director, and stockholder of Farber Partners and Serien Therapeutics. JDR and JMG are inventors of patents in the area of folate metabolism held by Princeton University. 1 bioRxiv preprint doi: https://doi.org/10.1101/2020.09.05.284521; this version posted September 7, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Abstract Mammalian cells require activated folates to generate nucleotides for growth and division. The most abundant circulating folate species is 5-methyl tetrahydrofolate (5-methyl- THF), which is used to synthesize methionine from homocysteine via the cobalamin-dependent enzyme methionine synthase (MTR).
    [Show full text]
  • L-Carnitine, Mecobalamin and Folic Acid Tablets) TRINERVE-LC
    For the use of a Registered Medical Practitioner or a Hospital or a Laboratory only (L-Carnitine, Mecobalamin and Folic acid Tablets) TRINERVE-LC 1. Name of the medicinal product Trinerve-LC Tablets 2. Qualitative and quantitative composition Each film- coated tablets contains L-Carnitine…………………….500 mg Mecobalamin……………….1500 mcg Folic acid IP…………………..1.5mg 3. Pharmaceutical form Film- coated tablets 4. Clinical particulars 4.1 Therapeutic indications Vitamin and micronutrient supplementation in the management of chronic disease. 4.2 Posology and method of administration For oral administration only. One tablet daily or as directed by physician. 4.3 Contraindications Hypersensitivity to any constituent of the product. 4.4 Special warnings and precautions for use L-Carnitine The safety and efficacy of oral L-Carnitine has not been evaluated in patients with renal insufficiency. Chronic administration of high doses of oral L-Carnitine in patients with severely compromised renal function or in ESRD patients on dialysis may result in accumulation of the potentially toxic metabolites, trimethylamine (TMA) and trimethylamine-N-oxide (TMAO), since these metabolites are normally excreted in the urine. Mecobalamin Should be given with caution in patients suffering from folate deficiency. The following warnings and precautions suggested with parent form – vitamin B12 The treatment of vitamin B12 deficiency can unmask the symptoms of polycythemia vera. Megaloblastic anemia is sometimes corrected by treatment with vitamin B12. But this can have very serious side effects. Don’t attempt vitamin B12 therapy without close supervision by healthcare provider. Do not take vitamin B12 if Leber’s disease, a hereditary eye disease.
    [Show full text]
  • Structural Basis for the Inhibition of Human 5,10-Methenyltetrahydrofolate Synthetase by N10-Substituted Folate Analogues
    Published OnlineFirst September 8, 2009; DOI: 10.1158/0008-5472.CAN-09-1927 Experimental Therapeutics, Molecular Targets, and Chemical Biology Structural Basis for the Inhibition of Human 5,10-Methenyltetrahydrofolate Synthetase by N10-Substituted Folate Analogues Dong Wu,1 Yang Li,1 Gaojie Song,1 Chongyun Cheng,1 Rongguang Zhang,2 Andrzej Joachimiak,2 NeilShaw, 1 and Zhi-Jie Liu1 1National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China and 2Structural Biology Center, Argonne National Laboratory, Argonne, Illinois Abstract of the one-carbon metabolic network, dihydrofolate reductase, 5,10-Methenyltetrahydrofolate synthetase (MTHFS) regulates which catalyzes the conversion of dihydrofolate to tetrahydrofolate. the flow of carbon through the one-carbon metabolic network, Similarly, colorectal and pancreatic cancers are routinely treated by which supplies essential components for the growth and administration of the pyrimidine analogue 5-fluorouracil (5-FU), proliferation of cells. Inhibition of MTHFS in human MCF-7 which inhibits thymidylate synthase (TS; ref. 6). TS converts breast cancer cells has been shown to arrest the growth of deoxyuridylate to deoxythymidylate using a carbon donated by cells. Absence of the three-dimensional structure of human 5,10-methylenetetrahydrofolate (Fig. 1). This conversion is essential MTHFS (hMTHFS) has hampered the rational design and for DNA synthesis (7). To increase the clinical efficiency of the optimization of drug candidates. Here, we report the treatment, 5-formyltetrahydrofolate is often coadministered along structures of native hMTHFS, a binary complex of hMTHFS with 5-FU. The beneficial effect of administering 5-formyltetrahy- with ADP, hMTHFS bound with the N5-iminium phosphate drofolate during the therapy with 5-FU is a consequence of the reaction intermediate, and an enzyme-product complex of 5,10-methenyltetrahydrofolate synthetase (MTHFS)–catalyzed hMTHFS.
    [Show full text]
  • Considerations for Crop Biofortification
    fpls-09-00443 April 4, 2018 Time: 16:15 # 1 REVIEW published: 06 April 2018 doi: 10.3389/fpls.2018.00443 Toward Eradication of B-Vitamin Deficiencies: Considerations for Crop Biofortification Simon Strobbe and Dominique Van Der Straeten* Laboratory of Functional Plant Biology, Department of Biology, Ghent University, Ghent, Belgium ‘Hidden hunger’ involves insufficient intake of micronutrients and is estimated to affect over two billion people on a global scale. Malnutrition of vitamins and minerals is known to cause an alarming number of casualties, even in the developed world. Many staple crops, although serving as the main dietary component for large population groups, deliver inadequate amounts of micronutrients. Biofortification, the augmentation of natural micronutrient levels in crop products through breeding or genetic engineering, is a pivotal tool in the fight against micronutrient malnutrition (MNM). Although these approaches have shown to be successful in several species, a more extensive knowledge of plant metabolism and function of these micronutrients is required to refine and improve biofortification strategies. This review focuses on the relevant B-vitamins Edited by: (B1, B6, and B9). First, the role of these vitamins in plant physiology is elaborated, Alexander Arthur Theodore Johnson, as well their biosynthesis. Second, the rationale behind vitamin biofortification is University of Melbourne, Australia illustrated in view of pathophysiology and epidemiology of the deficiency. Furthermore, Reviewed by: advances in biofortification, via metabolic engineering or breeding, are presented. Finally, Francesco Di Gioia, University of Florida, United States considerations on B-vitamin multi-biofortified crops are raised, comprising the possible Aymeric Goyer, interplay of these vitamins in planta.
    [Show full text]
  • Recombinant Human Dihydrofolate Reductase/DHFR
    Recombinant Human Dihydrofolate Reductase/DHFR Catalog Number: 8456-DR DESCRIPTION Source E. coli­derived Met1­Asp187, with a C­terminal 6­His tag Accession # P00374 N­terminal Sequence Met1 & Val2 Analysis Predicted Molecular 22 kDa Mass SPECIFICATIONS SDS­PAGE 23 kDa, reducing conditions Activity Measured by the reduction of dihydrofolic acid (DHF). The specific activity is >5,500 pmol/min/μg, as measured under the described conditions. Endotoxin Level <1.0 EU per 1 μg of the protein by the LAL method. Purity >95%, by SDS­PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane. Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl, Glycerol, Brij­35 and DTT. See Certificate of Analysis for details. Activity Assay Protocol Materials l Assay Buffer: 50 mM MES, 25 mM Tris, 100 mM NaCl, 25 mM Ethanolamine, 2 mM DTT l Recombinant Dihydrofolate Reductase/DHFR (rhDHFR) (Catalog # 8456­DR) l β­Nicotinamide adenine dinucleotide phosphate reduced, tetrasodium salt (β­NADPH) (Sigma, Catalog # N7505), 10 mM stock in deionized water l Dihydrofolic acid (DHF) (Sigma, Catalog # D7006), 10 mM stock in Assay Buffer + 4.5 mM NaOH l 96­well Clear Plate (Catalog # DY990) l Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent Assay 1. Dilute rhDHFR to 1 μg/mL in Assay Buffer. 2. Prepare a Substrate Mixture containing 0.2 mM DHF and 0.25 mM β­NADPH in Assay Buffer. 3. Load 50 μL of 1 μg/mL rhDHFR into a plate, and start the reaction by adding 50 μL of Substrate Mixture.
    [Show full text]
  • Oral Immunosuppressive Drugs in the Treatment of Atopic Dermatitis
    Oral immunosuppressive drugs in the treatment of atopic dermatitis IMPROVING PERFORMANCE AND SAFETY FLOOR M. GARRITSEN Colofon Oral immunosuppressive drugs in the treatment of atopic dermatitis Improving performance and safety Thesis with a summary in Dutch, Utrecht University, the Netherlands © Floor Garritsen, 2018 No part of this thesis may be reproduced, stored or transmitted, in any form or by any means, without prior permission of the author. ISBN: 978-90-393-6931-9 Cover design and layout: Die Jongens Printed and published by: Proefschriftmaken.nl Oral immunosuppressive drugs in the treatment of atopic dermatitis: improving performance and safety Orale immunosuppressiva bij de behandeling van constitutioneel eczeem: het verbeteren van effectiviteit en veiligheid (met een samenvatting in het Nederlands) Proefschrift ter verkrijging van de graad van doctor aan de Universiteit Utrecht op gezag van de rector magnificus, prof. dr. G.J. van der Zwaan, ingevolge het besluit van het college voor promoties in het openbaar te verdedigen op donderdag 8 februari 2018 des middags te 2.30 uur door Floralie Maria Garritsen geboren op 22 oktober 1986 te Beek en Donk Promotor: Prof. dr. C.A.F.M. Bruijnzeel-Koomen Copromotoren: Dr. M.S. de Bruin-Weller Dr. M.P.H. van den Broek Table of contents Chapter 1 General introduction 7 Part I – Qualitative and quantitative exploration of oral immunosuppressive drug use in the Netherlands Chapter 2 Ten years experience with oral immunosuppressive treatment 23 in adult patients with atopic dermatitis in two
    [Show full text]
  • BIOPAR DELTA-FORTE- 1 Nf Units, 50 Mcg Cbl, 2.5 Mg F-Thf, 1
    BIOPAR DELTA-FORTE- 1 nf units, 50 mcg cbl, 2.5 mg f-thf, 1 mg pteglu-, 7 mg me- thf capsule Jaymac Pharmacueticals LLC Disclaimer: This drug has not been found by FDA to be safe and effective, and this labeling has not been approved by FDA. For further information about unapproved drugs, click here. ---------- BioparTM delta-FORTE DESCRIPTION [1 NF Units] [50 mcg CBl] [2.5 mg F-THf] [1 mg PteGlu-] [7 mg Me-THf] Prescription Hematopoietic Preparation For Therapeutic Use Multiphasic Softgels (30ct carton) NDC 64661-793-30 Rx ONLY BioparTMdelta-FORTE is an orally administered prescription hematopoietic preparation for therapeutic use formulated for adult macrocytic anemia patients – specifically including pernicious anemia patients, ages 12 and up, who are under specific direction and monitoring of cobalamin and folate status by a physician. A recent study+ suggested that BioparTM delta-FORTE was effective in lowering homocysteine levels in patients that are positive for MTHFR (methylenetetrahydrofolate reductase polymorphism). BioparTM delta-FORTE may be taken by women of childbearing age. BioparTM delta- FORTE may be taken by geriatric patients where compliance is an issue. + ClinicalTrials.gov identifier: NCT02709668, Correlation of Clinical Response With Homocysteine Reduction During Therapy With Reduced B Vitamins in Patients with MDD Who Are Positive for MTHFR C677T or A1298C Polymorphism. INGREDIENTS Cobalamin-intrinsic factor concentrate (non-inhibitory)1 .............................................................................................
    [Show full text]
  • GRAS Notice 915, Calcium L-Methylfolate
    GRAS Notice (GRN) No. 915 https://www.fda.gov/food/generally-recognized-safe-gras/gras-notice-inventory Form Approved: 0MB No. 0910-0342; Expiration Date: 09/30/2019 (See last naae for 0MB Statement) FDA USE ONLY , 'GRN NUM~ER QATE OF. RECEIF!T DEPARTMENT OF HEALTH AND HUMAN SERVICES -- - ESTIMATED DAlllY·INTAKE fNTENDED USE FQ~ INTERNET Food and Drug Administration Ii .. GENERALLY RECOGNIZED AS SAFE -- 'NAME F:OR INTERNET (GRAS) NOTICE (Subpart E of Part 170) - - ,~ Y.WORDS I l I ~ .. Transmit completed form and attachments electronically via the Electronic Submission Gateway (see Instructions); OR Transmit completed form and attachments in paper format or on physical media to: Office of Food Additive Safety (HFS-200), Center for Food Safety and Applied Nutrition, Food and Drug Administration,5001 Campus Drive, College Park, MD 20740-3835. SECTION A- INTRODUCTORY INFORMATION ABOUT THE SUBMISSION 1. Type of Submission (Check one) ~ New D Amendment to GRN No. D Supplement to GRN No. 2. [gl All electronic files included in this submission have been checked and found to be virus free. (Check box to verify) 3 Mosl recent presuom1ss1on meeung (" any) w1m FDA on the subject substance (yyyylmmldd): 12/17/2018 4 For Amendments or Supplements: Is your (Check one) amendment or supplement submitted in D Yes If yes, enter the date of response lo a communication from FDA? D No communication (yyyy/mmldd): ______ SECTION B - INFORMATION ABOUT THE NOTIFIER Name of Contact Person Position or Title William Turney Head of Regulatory Affairs NA Organization (if applicable) 1 a. Notifier DSM Nutritional Products Mailing Address (number and street) 45 Waterview Blvd.
    [Show full text]
  • The Changing Faces of Vitamin B12 and Folic Acid – Part Two: Integration of Symptoms, and Genetic Testing to Select the Best Drugs for Your Patient
    The Changing Faces of Vitamin B12 and Folic Acid – Part two: Integration of symptoms, and genetic testing to select the best drugs for your patient By: Dan Carter, ND The symptoms of vitamin B12 deficiency range from mild to severe, and in many cases are not specific enough to confirm a diagnosis. Symptoms can include diarrhea or constipation, fatigue, light headedness on standing, poor appetite, pallor, difficulty concentrating, shortness of breath on exertion, glossitis. Neurologic symptoms can appear after chronic deficiency and involve changes in mental status, depression, peripheral neuropathy, loss of balance. [Anemia – B12 deficiency. http://www.nlm.nih.gov/medlineplus/ency/article/000574.htm] Some of the symptoms of folate deficiency overlap with B12, including fatigue and glossitis, but others such as accelerated graying of hair, oral ulcers, and poor growth rates do not. [Folate deficiency. http://www.nlm.nih.gov/medlineplus/ency/article/000354.htm] The indicated blood tests for B12 deficiency are complete blood count (CBC), reticulocyte count, serum LDH and vitamin B12, while those for folate deficiency are CBC, serum and RBC folate. Other useful tests are serum homocysteine and methylmalonic acid (MMA). Serum homocysteine will be high with both deficiencies, while only MMA will be increased in B12 deficiency. [Vitamin B12 and Folate. http://labtestsonline.org/understanding/analytes/vitamin-b12/tab/test] As genetic testing has become more widely available, specific analysis of known single nucleotide polymorphisms (SNP) is increasingly accepted as an aid to diagnosis. In the case of folate metabolism several SNP have been identified as significant; the first SNP to gain widespread attention affected the MTHFR gene.
    [Show full text]
  • Simulation of Food Folate Digestion and Bioavailability of an Oxidation Product of 5-Methyltetrahydrofolate
    Article Simulation of Food Folate Digestion and Bioavailability of an Oxidation Product of 5-Methyltetrahydrofolate Christiane Ringling 1 and Michael Rychlik 1,2,* 1 Division BIOANALYTIK Weihenstephan, Research Center for Nutrition and Food Sciences (Z. I. E. L.), Technical University of Munich, Alte Akademie 10, D-85350 Freising, Germany; [email protected] 2 Chair of Analytical Food Chemistry, Technical University of Munich, Alte Akademie 10, D-85350 Freising, Germany * Correspondence: [email protected]; Tel.: +49-8161-71-3746; Fax: +49-8161-71-4216 Received: 22 July 2017; Accepted: 28 August 2017; Published: 1 September 2017 Abstract: Generating bioavailability data from in vivo studies is time-consuming and expensive. In vitro simulation can help to investigate factors influencing bioavailability or facilitate quantifying the impact of such factors. For folates, an efficient deconjugation of polyglutamates to the corresponding monoglutamates is crucial for bioavailability and highly dependent on the food matrix. Therefore, the bioaccessibility of folates of different foodstuffs was examined using a simulated digestion model with respect to folate stability and the efficiency of deconjugation. For realistic simulated deconjugation, porcine brush border membrane was used during the phase of the simulated digestion in the small intestine. For a better understanding of folate behaviour during digestion, single folate monoglutamates were also investigated with this in vitro digestion model. The results for bioaccessibility were compared with data from a human bioavailability study. They support the idea that both stability and deconjugation have an influence on bioaccessibility and thus on bioavailability. Tetrahydrofolate is probably lost completely or at least to a high extent and the stability of 5-methyltetrahydrofolate depends on the food matrix.
    [Show full text]
  • Produktinformation Todavit
    SUMMARY OF PRODUCT CHARACTERISTICS 1. NAME OF THE MEDICINAL PRODUCT TODAVİT powder for solution for I.M./I.V. injection/ infusion Sterile 2. QUALITATIVE AND QUANTITATIVE COMPOSITION Active Substance: In each vial; Vitamin A (as Retinol palmitate) 3500 IU (2,06 mg) Vitamin D3 (Cholecalciferol) (made from sheep wool.) 220 IU (0,22 mg) Vitamin E (11,2 mg Vitamin E as acetate) 11,2 IU Vitamin C (Ascorbic acid ) 125 mg Vitamin B1 (Thiamine; 3,935 mg asThiamine HCl) 3,51 mg Vitamin B2 (Riboflavin; 5,26 mg Riboflavin sodium phosphate) 4,14 mg Vitamin B6 (Piridoxin; 5,5 mg as Piridoxin hydrochloride) 4,53 mg Vitamin B12 (Cyanocobalamin) 0,006 mg Folik acid 0,414 mg Pantothenic acid (16,15 mg asDexpanthenol) 17,25 mg D-Biotin 0,069 mg Nicotinamide 46 mg Excipients: Sodium hydroxide: q.s. Soy lecithin: 112.5 mg For a full list of excipients, see section 6.1 3. PHARMACEUTICAL FORM Yellow-orange colored lyophilized cake. Sterile. 4. CLINICAL PARTICULARS 4.1. Therapeutic indications In accordance with the daily needs of adults and children over 11 years of age,it is indicated when oral intake is contraindicated or inadequate (malnutrition, gastrointestinal malabsorption, parenteral nutrition, etc.) or when it is not possible, intravenously. 1/15 4.2. Posology and method of administration Posology / Application frequency and duration: In adults and children over 11 years old, the daily dose is 1 vial. In special cases with increased nutritional requirements (severe burns, etc.), high doses such as 2-3 vials daily can be used. Method of Application: It is used intravenously or intramuscularly.
    [Show full text]
  • (12) Patent Application Publication (10) Pub. No.: US 2002/0035087 A1 Barclay (43) Pub
    US 20020035087A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2002/0035087 A1 Barclay (43) Pub. Date: Mar. 21, 2002 (54) B COMPLEX VITAMIN COMPOSITIONS (57) ABSTRACT THAT PROTECT AGAINST CELLULAR The present invention relates generally to the use of Vitamin DAMAGE CAUSED BY ULTRAVIOLET B12 (cobalamin or cyanocobalamin) alone or in combina LIGHT tion with other photoprotective agents, including Specifically other vitamins such as vitamin B9 (folic acid or folate) and Vitamin B3 (niacin or niacinamide), or any chemical deriva (76) Inventor: Barry J. Barclay, St. Albert (CA) tive of these Vitamins and their Salts, as a filter to protect cells against the damaging effects of ultraviolet (UV) light. The invention is, in one aspect, a method of reducing the rate Correspondence Address: of UV damage to cells exposed to a UV light source, by Kevin S. Lemack treating the celis with the Vitamin composition, either alone Nields & Lemack or in combination with other photoprotective agents. Other 176 E. Main Street - Suite 8 aspects of the invention are compositions comprising effec Westboro, MA 01581 (US) tive amounts of vitamin B12 alone or in combination with other photoprotective agents including Vitamin B9 and Vita min B3 and a pharmaceutically-acceptable carrier, that are (21) Appl. No.: 09/900,064 useful in protecting cells, particularly Skin cells, against the burning, genotoxic (mutagenic and carcinogenic), immuno (22) Filed: Jul. 6, 2001 Suppressive and photoaging effects of UV light, especially Sunlight. The invention has application as a UV light filter in (30) Foreign Application Priority Data oral preparations including tablets and drinks, topical creams, lotions, sprays, wipes and cosmetics.
    [Show full text]