DOCSLIB.ORG

Biosynthetic Production and Structural Studies of Glycolipids and Large Fragments of G-Protein Coupled Receptors

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Biosynthetic Production and Structural Studies of Glycolipids and Large Fragments of G-Protein Coupled Receptors Zurich Open Repository and Archive University of Zurich Main Library Strickhofstrasse 39 CH-8057 Zurich www.zora.uzh.ch Year: 2010 Biosynthetic production and structural studies of glycolipids and large fragments of G-protein coupled receptors Kumaran, Sowmini Posted at the Zurich Open Repository and Archive, University of Zurich ZORA URL: https://doi.org/10.5167/uzh-42491 Dissertation Published Version Originally published at: Kumaran, Sowmini. Biosynthetic production and structural studies of glycolipids and large fragments of G-protein coupled receptors. 2010, University of Zurich, Faculty of Science. !" !"&'()'"'()* +,'$" #+0&.+1'*,'$/-02(.$/.")$( 0(12%' /"2".$+0&.( 30-7)+8-079)&'!( 0(172* $)"&(8 ,2/).(9)*)5' *!! :"$$)-$0'" # ;,- ?-20&4,&7(.)-(&0',-="$$)&$'(03$2"*()&(: >$ *=B-.) A:-D($'D(&0'DE D *7)2)4$(.)- E0'()90$"$'(7&0',-="$$)&$'(03$2"*()&(80A,2'F$ .)- I&"F)-$"$F$(HB-"'( D # " !%&'&!()%'('$ 0/$ ,'-&.$! ((((((,- 6 $" #$A 9"$)) ,( -.!2(.!04&5.(!3.(7.!89 (7#89!)'-!:.#8&$;!-.(!2&77.(8'8# $< !,( -.!2(.!= >$!B 4#$7 $ HB-"'(I(LMNM Table of Contents Abbreviation 1 Summary 5 Zusamenfassung 8 Chapter 1 11 1.0 Introduction 11 1.1 Chemistry of protein–carbohydrate interactions 12 1.1.1 Hydrogen bonds 13 1.1.2 Hydrophobic interactions 13 1.1.3 Electrostatic interactions 13 1.1.4 Role of water 14 1.2 Tools for analyzing carbohydrate protein interactions 14 1.2.1 NMR approaches to study carbohydrate–protein Interactions 14 1.2.2 NMR parameters that yield information on protein ligand interactions physiological functions of Y receptors 16 1.2.2.1 Chemical shift 16 1.2.2.2 Relaxation parameters 16 1.2.2.3. NOEs 17 1.2.2.4. Magnetization transfer rates 18 1.2.2.5 Aspects of binding affinity and timescale for detection of NMR parameters 18 1.2.3 Experiments based on chemical shift perturbation 21 1.2.4 Saturation transfer difference (STD) nuclear magnetic resonance experiments to detect ligand binding 21 1.2.5 Water LOGSY: water-ligand observation with gradient spectroscopy 23 1.2.6 Transferred intramolecular nuclear overhauser effects (trNOEs) 23 1.2.6.1 The transferred ROESY experiment 25 1.2.7 Isotope filtering/editing using labeled ligands 25 1.2.8 Spin labeling 27 1.3 Other methods to investigate protein-carbohydrate interactions 28 1.3.1 Molecular dynamics simulations 28 1.3.2 X-Ray crystallography 29 1.3.3 Fluorescence spectrometry in studies of carbohydrate-protein interactions 30 1.3.3.1 Polarisation and anistropy 31 1.3.3.2 FRET 31 1.3.4 Calorimetric experiments 31 1.3.4.1 Isothermal titration calorimetry (ITC) 31 1.3.4.2. Differential scanning calorimetry (DSC) 33 1.3.5 Surface plasmon resonance 33 1.4 Molecules under study 34 1.4.1 Lipopolysaccharides 34 1.4.2 Structure and different types of LPS 35 1.4.3 Structural requirements for the bioactivity of LPS 39 1.4.4 Biosynthesis of LPS 40 1.4.5 LPS signaling pathways 41 1.4.6 Key players of LPS signaling pathway - Endotoxin-binding proteins 43 1.4.6.1 LBP 43 1.4.6.2 BPI 43 1.4.6.3 CD14 44 1.4.6.4 Structure of CD14 45 1.4.6.5 Toll-like receptors 46 1.4.7 LPS responsive cells 47 1.4.7.1 Monocytes 47 1.4.7.2 Polymorphonuclear leukocytes (PMN) 47 1.4.7.3 B- and T-lymphocytes 48 1.4.7.4 Vascular cells, epithelial cells 48 1.4.8 Strategies of endotoxin antagonism 48 1.4.8.1 Anti-endotoxin antibodies 48 1.4.8.2 LPS-binding proteins 49 1.4.8.3 Anti-LPS peptides 49 1.4.8.4 Antimicrobial peptides 49 1.4.9 Polymyxins 50 1.4.9.1 Structure of polymyxins 51 1.4.9.2 Types of polymyxins 51 1.4.9.3 Mechanism of action 53 1.5 Scope of our study 53 1.6 References 55 Chapter 2 2.0 Introduction 62 2.1 A Brief History of GPCRs 63 2.2 GPCR signaling 64 2.3 Significance of GPCRs 66 2.4 Classification of GPCRs 67 2.5 Expression of GPCRs 70 2.6 Current state of structural studies of GPCRs 71 2.6.1 General structure 72 2.6.1.1 Why Seven TMs? 72 2.6.2 The Rhodopsin Structure 73 2.6.2.1 Transmembrane region 74 2.6.2.2 Retinal binding site 75 2.6.2.3 Cytoplasmic surface 76 2.6.2.4 Intramolecular interactions and GPCR activation 76 2.6.3 Opsin/Rhodopsin in the activated state 78 2.6.4 Adrenergic receptors 79 2.6.4.1 !2-adrenergic G-protein coupled receptor 79 2.6.4.2 !1-adrenergic G-protein coupled receptor 81 2.6.4.3 Architecture of the Human A2A Adenosine receptor 82 2.6.5 Comparative analysis of GPCR crystal structures 84 2.7 GPCR oligomerization 87 2.7.1 GPCR studies in fragments 87 2.7.2 Helix –helix interactions 89 2.7.3 A membrane protein folding model 89 2.7.4 Membrane mimetics 90 2.7.4.1 Organic solvents 91 2.7.4.2 Micelles 91 2.7.4.3 Bicelles 92 2.8 Neuropeptide receptors: Aim and scope of the investigation 92 2.9 References 97 Chapter 3 Expression of sCD14 (1-152) in E.coli and Pichia pastoris 104 3.0 Introduction 104 3.1 LPS 104 3.2 CD14 106 3.3 Summary of expression of sCD14(1-152) 107 3.3.1 Recombinant expression of sCD14(1-152) in E.coli 107 3.3.2 Recombinant protein expression in Pichia Pastoris 116 3.3.2.1 The Pichia expression vector 116 3.3.2.2 How it works 117 3.3.2.3 Experimental overview 118 3.3.2.4 Expression of CD14 in Pichia pastoris 118 3.4 Outlook 120 3.5 References 120 Chapter 4 Interactions of lipopolysaccharide and polymyxin studied by NMR spectroscopy 122 4.0 Introduction 122 4.1 Experimental procedures 124 4.1.1 Materials 124 4.1.2 Production of 13C-labelled LPS from the E. coli strain D31m4 124 4.1.3 Production of 13C,15N-labelled PMX-M from P. kobensis M 125 4.1.4 NMR spectroscopy 126 4.1.5 Molecular dynamics calculations 127 4.2 Results 128 4.2.1 Production and purification of 13C-labeled LPS 128 4.2.2 Assignment of LPS and polymyxin resonances 129 4.2.3 Topology of LPS and polymyxin insertion into the DPC micelle 130 4.2.4 Interactions of LPS with various types of polymyxins 131 4.3 Discussions 135 4.4 Acknowledgements 138 4.5 References 138 4.6 Supplementary materials 143 Chapter 5 Studies of the structure of the N-terminal domain from the Y4 receptor, a G-protein coupled receptor and its interaction with hormones from the NPY family 155 5.0 Introduction 155 5.1 Results 157 5.1.1 Recombinant production of N-Y4 157 5.1.2 The structure of N-Y4 158 5.1.3 Topology of membrane-association 161 5.1.4 Immobilizing the N-terminus on the membrane 162 5.1.5 Interaction between N-Y4 and neuropeptides from the NPY family 162 5.2 Discussions 166 5.3 Conclusions 169 5.4 Materials and methods 170 5.4.1 Plasmid construction, expression and purification of N-Y4 171 5.4.2 Synthesis and purification of the neuropeptides and of unlabelled N-terminal fragments 172 5.4.3 Dodecylphosphoethanolamine coupling to carboxyl terminus of N-Y4 173 5.4.4 NMR experiments 173 5.4.5 Membrane-association topology using spin labels 174 5.4.6 Surface plasmon resonance (SPR) studies 174 5.5 Acknowledgement 175 5.6 References 175 5.7 Supplementary materials 177 Chapter 6 Properties of the N-terminal domains from Y receptors probed by NMR spectroscopy 187 6.0 Introduction 187 6.1 Results 190 6.1.1 Expression of N-terminal domains in isotopically-labeled form 190 6.1.2 Assignment of chemical shifts 193 6.1.3 Screening structural properties by 15N relaxation and CD spectroscopy 193 6.1.4 Structures of the N-terminal domains in presence of micelles 195 6.1.5 Interaction studies with neuropeptides from the NPY family 197 6.2 Discussions 200 6.3 Materials and methods 203 6.3.1 Materials 203 6.3.2 Expression and purification of N-terminal domains 203 6.3.3 NMR spectroscopy 205 6.4 Acknowledgements 206 6.5 References 206 6.6 Supplementary materials 211 Chapter 7 Expression of Double Transmembrane Fragments of Y4 Receptor, a Human GPCR, for Structural Studies by NMR Spectroscopy 222 7.0 Introduction 222 7.1 Materials and methods 227 7.1.1 Cloning and mutagenesis to generate double transmembrane constructs of human Y4 receptor into pLC01 vector 227 7.1.2 Protein expression and purification of double transmembrane fragments 229 7.1.3 CNBr cleavage 230 7.1.4 Production of N-His6-3C protease 231 7.1.5 Screening of cleavage conditions for Y4 Trp!LE-TM6-TM7-C by the C3 protease 231 7.1.6 NMR sample preparation 232 7.2 Results and discussion 232 7.2.1 Design and generation of double transmembrane constructs of human Y4- receptor into pLC01 vector 232 7.2.2 The choice of the expression vector systems 233 7.2.3 Selections of E.coli host strains 235 7.2.4 Optimization of inducible expression and purification 237 7.2.5 Optmization of expression in deuterated growth medium 240 7.2.6 Optimization of the cleavage of the fusion protein 241 7.2.7 Detergent screening 245 7.2.8 Construction of larger transmembrane fragments of Y4 receptor 249 7.3 Conclusions 250 7.4 References 252 Curriculum Vitae 254 Acknowledgements 258 Abbreviation AM adrenomedullin AMP Antimicrobial peptide AR adrenergic receptor CARA computer aided resonance assignment CCR2 chemokine C-C motif receptor 2 CCR5 chemokine C-C motif receptor 5 CGRP calcitonin generelated peptide CD circular dichroism CD14 Cluster of differentiation 14 cDNA complementary deoxyriobonucleic acid CHAPS 3-(3-Cholamidopropyl)dimethylammonio)-1-Propanesulfonic Acid CMC critical micelle concentration CNS central nervous system CV column volume CX3C chemokine C-X3-C motif receptor Dab Diamino butyric acid DCM dichloromethane DDM dodecyl-!-d-maltoside DHPC 1,2-Diheptanoyl-sn-Glycero-3-Phosphocholine DIEA diisopropylethylamine DMF dimethyl formamide DPC dodecylphosphocholine Doxyl (4,4-dimethyl-3-oxazolidine-N-oxyl) DTT 1,4-dithiothreitol ECL extracellular loop E.coli Escherichia coli EDTA ethylenediamine tetraacetic acid ELISA enzyme linked immunostaining assay 1 ESI-MS electrospray ionization
Load more

Recommended publications
  • Neotrance and the Psychedelic Festival DC
    Neotrance and the Psychedelic Festival GRAHAM ST JOHN UNIVERSITY OF REGINA, UNIVERSITY OF QUEENSLAND Abstract !is article explores the religio-spiritual characteristics of psytrance (psychedelic trance), attending speci"cally to the characteristics of what I call neotrance apparent within the contemporary trance event, the countercultural inheritance of the “tribal” psytrance festival, and the dramatizing of participants’ “ultimate concerns” within the festival framework. An exploration of the psychedelic festival offers insights on ecstatic (self- transcendent), performative (self-expressive) and re!exive (conscious alternative) trajectories within psytrance music culture. I address this dynamic with reference to Portugal’s Boom Festival. Keywords psytrance, neotrance, psychedelic festival, trance states, religion, new spirituality, liminality, neotribe Figure 1: Main Floor, Boom Festival 2008, Portugal – Photo by jakob kolar www.jacomedia.net As electronic dance music cultures (EDMCs) flourish in the global present, their relig- ious and/or spiritual character have become common subjects of exploration for scholars of religion, music and culture.1 This article addresses the religio-spiritual Dancecult: Journal of Electronic Dance Music Culture 1(1) 2009, 35-64 + Dancecult ISSN 1947-5403 ©2009 Dancecult http://www.dancecult.net/ DC Journal of Electronic Dance Music Culture – DOI 10.12801/1947-5403.2009.01.01.03 + D DC –C 36 Dancecult: Journal of Electronic Dance Music Culture • vol 1 no 1 characteristics of psytrance (psychedelic trance), attending specifically to the charac- teristics of the contemporary trance event which I call neotrance, the countercultural inheritance of the “tribal” psytrance festival, and the dramatizing of participants’ “ul- timate concerns” within the framework of the “visionary” music festival.
    [Show full text]
  • Application of High Resolution Mass Spectrometry for the Screening and Confirmation of Novel Psychoactive Substances Joshua Zolton Seither [email protected]
    Florida International University FIU Digital Commons FIU Electronic Theses and Dissertations University Graduate School 4-25-2018 Application of High Resolution Mass Spectrometry for the Screening and Confirmation of Novel Psychoactive Substances Joshua Zolton Seither [email protected] DOI: 10.25148/etd.FIDC006565 Follow this and additional works at: https://digitalcommons.fiu.edu/etd Part of the Chemistry Commons Recommended Citation Seither, Joshua Zolton, "Application of High Resolution Mass Spectrometry for the Screening and Confirmation of Novel Psychoactive Substances" (2018). FIU Electronic Theses and Dissertations. 3823. https://digitalcommons.fiu.edu/etd/3823 This work is brought to you for free and open access by the University Graduate School at FIU Digital Commons. It has been accepted for inclusion in FIU Electronic Theses and Dissertations by an authorized administrator of FIU Digital Commons. For more information, please contact [email protected]. FLORIDA INTERNATIONAL UNIVERSITY Miami, Florida APPLICATION OF HIGH RESOLUTION MASS SPECTROMETRY FOR THE SCREENING AND CONFIRMATION OF NOVEL PSYCHOACTIVE SUBSTANCES A dissertation submitted in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY in CHEMISTRY by Joshua Zolton Seither 2018 To: Dean Michael R. Heithaus College of Arts, Sciences and Education This dissertation, written by Joshua Zolton Seither, and entitled Application of High- Resolution Mass Spectrometry for the Screening and Confirmation of Novel Psychoactive Substances, having been approved in respect to style and intellectual content, is referred to you for judgment. We have read this dissertation and recommend that it be approved. _______________________________________ Piero Gardinali _______________________________________ Bruce McCord _______________________________________ DeEtta Mills _______________________________________ Stanislaw Wnuk _______________________________________ Anthony DeCaprio, Major Professor Date of Defense: April 25, 2018 The dissertation of Joshua Zolton Seither is approved.
    [Show full text]
  • Inner Visions: Sacred Plants, Art and Spirituality
    AM 9:31 2 12/10/14 2 224926_Covers_DEC10.indd INNER VISIONS: SACRED PLANTS, ART AND SPIRITUALITY Brauer Museum of Art • Valparaiso University Vision 12: Three Types of Sorcerers Gouache on paper, 12 x 16 inches. 1989 Pablo Amaringo 224926_Covers_DEC10.indd 3 12/10/14 9:31 AM 3 224926_Text_Dec12.indd 3 12/12/14 11:42 AM Inner Visions: Sacred Plants, Art and Spirituality • An Exhibition of Art Presented by the Brauer Museum • Curated by Luis Eduardo Luna 4 224926_Text.indd 4 12/9/14 10:00 PM Contents 6 From the Director Gregg Hertzlieb 9 Introduction Robert Sirko 13 Inner Visions: Sacred Plants, Art and Spirituality Luis Eduardo Luna 29 Encountering Other Worlds, Amazonian and Biblical Richard E. DeMaris 35 The Artist and the Shaman: Seen and Unseen Worlds Robert Sirko 73 Exhibition Listing 5 224926_Text.indd 5 12/9/14 10:00 PM From the Director In this Brauer Museum of Art exhibition and accompanying other than earthly existence. Additionally, while some objects publication, expertly curated by the noted scholar Luis Eduardo may be culture specific in their references and nature, they are Luna, we explore the complex and enigmatic topic of the also broadly influential on many levels to, say, contemporary ritual use of sacred plants to achieve visionary states of mind. American and European subcultures, as well as to contemporary Working as a team, Luna, Valparaiso University Associate artistic practices in general. Professor of Art Robert Sirko, Valparaiso University Professor We at the Brauer Museum of Art wish to thank the Richard E. DeMaris and the Brauer Museum staff present following individuals and agencies for making this exhibition our efforts of examining visual products arising from the possible: the Brauer Museum of Art’s Brauer Endowment, ingestion of these sacred plants and brews such as ayahuasca.
    [Show full text]
  • Research Chemicals Und Andere Neue Drogen
    Research Chemicals & andere „neue“ Drogen Impulsreferat auf der DO-Leitertagung am 08.04.2014 Dipl.-Psych. Marcus Breuer psycholog. Psychotherapeut Begriffsklärung • Crystal = Crystal Speed; kristallines Metamphetamin • ATS = Amphetamin Type Stimulants • NPS = New Psychoactive Substances • Badesalze = Szenebezeichnung, denkbar ungeeignet, weil sie nichts erklärt und außerdem verharmlost • Research Chemicals = diese Stoffe sind „Abfallprodukte“ aus der Medikamentenentwicklung oder aber gezielte Veränderungen bekannter psychoaktiver Substanzen • Legal Highs = viele dieser neuen Stoffe sind bzw. waren zunächst legal (nicht dem BtmG unterstellt) • Kräutermischungen = Spice & Co., rauchbare synthetische Cannabinoide (viel toxischer als Cannabis) Wirkbogen der Partydrogen Entaktogene Ecstasy-MDMA MDEE Häufigkeit • ATS= Amphetamine Type Stimulants • Amphetamin (Speed) • XTC (MDMA; MDE; MDA) • Methamphetamin (Crystal) • Weltweit zusammen 26 million “regular users” vs: • Kokain 14 million reg. users • Heroin 16 million reg. users Nach WHO and UNODOC-Schätzungen (2010) ist Methamphetamin weltweit alleine die nach CANNABIS am häufigsten konsumierte illegale Substanz(18 Mio) (Nikotin und Alkohol sind Hauptproblem…..) Gesamtzahlen erstauffällige Konsumenten nach BKA: Trend 2000-2009 Drogentrends Exkurs: Research Chemicals I Neue Clubdrugs/Trenddrogen - fast alles RC-Drugs: • Substanzen, die zu Forschungszwecken genutzt werden und als solche „gekauft“ werden…. • RC-Substanzen (research chemicals) / Designerdrugs….. • Zeitweise nicht dem BTMG der jeweiligen
    [Show full text]
  • From Religious Ecstasy to Ecstasy Pills: a Symbolic and Performative Analysis of Electronic Music Festivals
    Relig. Soc. vol.2 no.se Rio de Janeiro 2006 From religious ecstasy to ecstasy pills: a symbolic and performative analysis of electronic music festivals Tiago Coutinho ABSTRACT This article looks to analyze the process through which eastern cosmological elements and religious practices acquire new meanings as they take on fresh uses in western festive contexts, as well as examine the symbolic and performative dimensions of the phenomenon in question. Electronic music festivals suggest a reading of eastern religious factors that serve as a reference to their ‘origin myth.’ The ethnographic data reveals a new form of obtaining ecstasy via music, performances, ‘natural’ ambients and altered states of consciousness. The appeal to transcendence is the direct result of western re-workings of Indian religious practices – especially those proposed by the spiritual leader Osho, who formed a cosmology based on fragments taken out of their ‘original’ context and given meaning in the life of those who adopt the Sannyasa lifestyle. Keywords: party, performance, psychoactive drugs, New Age spirituality Introduction Based on the ethnographic material contained in my master’s dissertation, this article explores a movement of resignification that transplants fragments of eastern religious systems to the festival contexts of young westerners. It looks to highlight the symbolic and performative dimensions of this phenomenon and its particular appeal to transcendence. The specific type of electronic music festival under study exemplifies the transposition from eastern practices to western capitalist contexts via facts and events occurring over the last thirty years that rework the meanings of one of the most important youth countercultures of the 20th century, the hippie movement.
    [Show full text]
  • Pharmacology Notebook 1 PDF (Searchable)
    The Shulgin Lab Books December 2012 May 2018 Pharmacology Lab Notes #1 (1960 - 1976) A Bit About This Document: While undertaking the work of investigating the chemistry and pharmacology of many varied psychoactive substances, Alexander “Sasha” Shulgin kept detailed notebooks. His documentation covered not only on his own personal research, but the research of friends and acquaintances. This book, the first of the “Pharmacology” series, represents mostly sub-acute work-ups of various substances and subjective responses by Shulgin and his research group. It covers the time frame of 1960 to 1976. The Creation of This Document: The project to undertake the transcribing of Shulgin’s Lab Books was started in 2008 by a team of volunteers and staff at Erowid, along with members of Team Shulgin. Various books were transcribed without a clear idea of how to present the information as a final product; eventually this format was chosen and a volunteer began work assembling the document. Each page was painstakingly transcribed from scanned images. All the hand-drawn “dirty pictures” (molecule drawings) and graphs were edited from the original scans and combined with drawn-in marks, outlines, and arrows to form this searchable PDF. Most of the names in this document have been redacted and pseudonyms put in their place. Names are presented as much as possible as they were in the original book, for example “Robert Thompson” is also “Robert”, “R.Thompson”, and “RT”. Initials are frequently used, and no two people share names or initials so the reader can keep track of who’s who. Words highlighted in yellow are words that the transcription team could not decipher.
    [Show full text]
  • Rage in Eden Records, Po Box 17, 78-210 Bialogard 2, Poland [email protected]
    RAGE IN EDEN RECORDS, PO BOX 17, 78-210 BIALOGARD 2, POLAND [email protected], WWW.RAGEINEDEN.ORG Artist Title Label HAUSCHKA ROOM TO EXPAND 130701/FAT CAT CD RICHTER, MAX BLUE NOTEBOOKS, THE 130701/FAT CAT CD RICHTER, MAX SONGS FROM BEFORE 130701/FAT CAT CD ASCENSION OF THE WAT NUMINOSUM 13TH PLANET RECORDS CD MINISTRY COVER UP 13TH PLANET RECORDS CD MINISTRY LAST SUCKER, THE 13TH PLANET RECORDS CD MINISTRY LAST SUCKER, THE 13TH PLANET RECORDS LTD MINISTRY RIO GRANDE BLOOD 13TH PLANET RECORDS CD MINISTRY RIO GRANDE DUB YA 13TH PLANET RECORDS CD PRONG POWER OF THE DAMAGER 13TH PLANET RECORDS CD REVOLTING COCKS COCKED AND LOADED 13TH PLANET RECORDS CD REVOLTING COCKS COCKTAIL MIXXX 13TH PLANET RECORDS CD BERNOCCHI, ERALDO/FE MANUAL 21ST RECORDS CD BOTTO & BRUNO/THE FA BOTTO & BRUNO/THE FAMILY 21ST RECORDS CD FLOWERS OF NOW INTUITIVE MUSIC LIVE IN COLOGNE 21ST RECORDS CD LOST SIGNAL EVISCERATE 23DB RECORDS CD SEVENDUST ALPHA 7 BROS RECORDS CD SEVENDUST CHAPTER VII: HOPE & SORROW 7 BROS RECORDS CD A BLUE OCEAN DREAM COLD A DIFFERENT DRUM MCD A BLUE OCEAN DREAM ON THE ROAD TO WISDOM A DIFFERENT DRUM CD B!MACHINE ALTERNATES AND REMIXES A DIFFERENT DRUM CD B!MACHINE EVENING BELL, THE A DIFFERENT DRUM CD B!MACHINE FALLING STAR, THE A DIFFERENT DRUM CD B!MACHINE MACHINE BOX A DIFFERENT DRUM BOX BLUE OCTOBER ONE DAY SILVER, ONE DAY GOLD A DIFFERENT DRUM CD BLUE OCTOBER UK INCOMING 10th A DIFFERENT DRUM 2CD CAPSIZE A PERFECT WRECK A DIFFERENT DRUM CD COSMIC ALLY TWIN SUN A DIFFERENT DRUM CD COSMICITY ESCAPE POD FOR TWO A DIFFERENT DRUM CD DIGNITY
    [Show full text]
  • UNCTAD Illicit Trade Forum Illicit Trade Report 2018
    UNCTAD Illicit Trade Forum 3rd to 4th February 2020 Room XXVI, Palais des Nations, Geneva Illicit Trade Report 2018 Contribution by World Customs Organization (WCO) The views expressed are those of the author and do not necessarily reflect the views of UNCTAD Illicit Trade Report 2018 www.wcoomd.org CONTENTS P. 2 FOREWORD P. 4 INTRODUCTION P. 6 Section 1. CULTURAL HERITAGE P.28 Section 2. DRUGS P. 88 Section 3. ENVIRONMENT P. 112 Section 4. IPR, HEALTH AND SAFETY P. 140 Section 5. REVENUE P. 174 Section 6. SECURITY P. 196 ABBREVIATIONS FOREWORD Illicit trafficking of different commodities continues to affect global seizures, themselves, cannot prevent or completely stop illicit trade, peace and security, destabilizing economies and threatening the they can serve as an active deterrent and raise the cost of illicit health and safety of populations. Disrupting illicit trade flows is a transactions. Moreover, enforcement measures tame illicit markets very complex, multi-stakeholder process, involving many law en- when they are a product of smart and well-researched policies, a forcement and other government agencies. The work done by all whole-of-government approach, and international cooperation. stakeholders, in 2018, dovetails with the World Customs Organiza- tion’s (WCO) theme for this specific year, which encouraged the pro- In 2018, a total of 154 countries reported their seizures to the WCO motion of “A Secure Business Environment for Economic Develop- through a specialized channel, namely the Customs Enforcement ment” – an apt theme in the context of illicit trade, given that illegal Network (CEN). Spanning reports on seizures of cultural objects by cross-border trade can cause financial losses for both legitimate 20 Customs administrations to 105 reporting seizures of drugs, the traders and governments, while impacting negatively on a host of CEN has accumulated a wealth of information that is analysed in other areas, such as governance and development objectives.
    [Show full text]
  • Neue Drogen – Neue Therapie? Crystal Speed, ATS &
    Neue Drogen – neue Therapie? Crystal Speed, ATS & NPS Arbeitsgruppe / Workshop auf der Buss-Jahrestagung am 12.03.2014 Dipl.-Psych. Marcus Breuer psycholog. Psychotherapeut Begriffsklärung • Crystal = Crystal Speed; kristallines Metamphetamin • ATS = Amphetamin Type Stimulants • NPS = New Psychoactive Substances • Badesalze = Szenebezeichnung, denkbar ungeeignet, weil sie nichts erklärt und außerdem verharmlost • Research Chemicals = diese Stoffe sind „Abfallprodukte“ aus der Medikamentenentwicklung oder aber gezielte Veränderungen bekannter psychoaktiver Substanzen • Legal Highs = viele dieser neuen Stoffe sind bzw. waren zunächst legal (nicht dem BtmG unterstellt) • Kräutermischungen = Spice & Co., rauchbare synthetische Cannabinoide (viel toxischer als Cannabis) Definition ATS • engl.: Amphetamine-Type-Stimulants • Amphetaminartige Substanzen, die eine ähnliche Ringstruktur haben wie das „klassische“ Amphetamin • Aber: unterschiedliche Wirkung….. Wirkbogen der Partydrogen Entaktogene Ecstasy-MDMA MDEE Häufigkeit • ATS= Amphetamine Type Stimulants • Amphetamin (Speed) • XTC (MDMA; MDE; MDA) • Methamphetamin (Crystal) • Weltweit zusammen 26 million “regular users” vs: • Kokain 14 million reg. users • Heroin 16 million reg. users Nach WHO and UNODOC-Schätzungen (2010) ist Methamphetamin weltweit alleine die nach CANNABIS am häufigsten konsumierte illegale Substanz(18 Mio) (Nikotin und Alkohol sind Hauptproblem…..) Gesamtzahlen erstauffällige Konsumenten nach BKA: Trend 2000-2009 Drogentrends ATS – Geschichte I - 1800 Ephedrin erstmalig
    [Show full text]
  • Us 2019 / 0142851 A1
    US 20190142851A1 ( 19) United States (12 ) Patent Application Publication ( 10) Pub . No. : US 2019 /0142851 A1 CHADEAYNE (43 ) Pub. Date : May 16 , 2019 ( 54 ) COMPOSITIONS COMPRISING A 2017 , provisional application No. 62/ 587, 410 , filed PSILOCYBIN DERIVATIVE AND A on Nov. 16 , 2017 , provisional application No . 62 /587 , CANNABINOID 395 , filed on Nov . 16 , 2017 . (71 ) Applicant: CaaMTech , LLC , Issaquah , WA (US ) Publication Classification (51 ) Int . Ci. ( 72 ) Inventor: Andrew R . CHADEAYNE , Issaquah , A61K 31/ 675 ( 2006 .01 ) WA (US ) A61K 31 / 4045 (2006 . 01 ) A61K 31/ 4406 ( 2006 .01 ) (21 ) Appl. No. : 16 / 192, 736 A61K 31/ 05 (2006 . 01) A61K 31 / 192 (2006 .01 ) ( 22 ) Filed : Nov . 15 , 2018 A61K 31 /353 ( 2006 . 01) (52 ) U . S . CI. Related U . S . Application Data CPC .. A61K 31/ 675 ( 2013 .01 ) ; A61K 31/ 4045 (60 ) Provisional application No . 62/ 613 , 360 , filed on Jan . (2013 . 01 ) ; A61K 31/ 353 (2013 .01 ) ; ACIK 3 , 2018 , provisional application No. 62 /609 , 115 , filed 31/ 05 (2013 . 01 ) ; A61K 31/ 192 (2013 . 01 ) ; on Dec . 21 , 2017 , provisional application No . 62/ 598 , A61K 31 /4406 ( 2013 . 01 ) 767, filed on Dec . 14 , 2017 , provisional application No . 62 /595 ,336 , filed on Dec . 6 , 2017 , provisional ( 57 ) ABSTRACT application No . 62 / 595 , 321 , filed on Dec . 6 , 2017 , This disclosure pertains to new compositions and methods provisional application No . 62 /593 ,021 , filed on Nov . comprising a psilocybin derivative . In one embodiment, the 30 , 2017 , provisional application No . 62 / 592, 320 , compositions disclosed herein are used for a method of filed on Nov . 29 , 2017 , provisional application No .
    [Show full text]
  • Natural Refrigerants
    cover_buch_090608 12.06.2008 19:24 Uhr Seite 1 Proklima International The replacement of R22 in the commercial and industrial refrigeration and air conditioning sector poses a big challenge for developing countries when designing and implementing the accelerated HCFC phase-out. This collec- tion of articles was compiled to highlight that natural refrigerants may be applied in various systems which previously used R22 and that they have proven to be energy-efficient, economically feasible and environmentally friendly replacements to HCFCs. The handbook contains 31 articles by individual authors covering the follow- ing issues: policies and legislation on F-gases and related issues (Part 1), safety concerns and means to overcome (Part 2), a technical assessment of natural refrigerants in different applications (air conditioning, commercial and industrial refrigeration and heat pumps) (Part 3), and case studies by manufacturers and end-users providing insights into market developments and examples of successful conversions to natural refrigerants (Part 4). PROKLIMA s t n a r Natural Refrigerants e g i Sustainable Ozone- and Climate-Friendly Alternatives to HCFCs r f e R l a r u t a N Deutsche Gesellschaft für Technische Zusammenarbeit (GTZ) GmbH – German Technical Cooperation – Programme Proklima Dag-Hammarskjöld-Weg 1-5 65760 Eschborn Sustainable Ozone- and Climate-Friendly Alternatives to HCFCs T + 49 6196 79 - 0 GTZ PROKLIMA F + 49 6196 79 - 6318 E [email protected] I www.gtz.de/proklima Proklima International Natural Refrigerants Sustainable Ozone- and Climate-Friendly Alternatives to HCFCs Published by: PROKLIMA is a programme of the Deutsche Gesellschaft für Technische Zusam- Deutsche Gesellschaft für Technische Zusammenarbeit (GTZ) GmbH menarbeit GmbH (GTZ).
    [Show full text]
  • General Anesthesia: a Literature Review - Induction, Mechanism, Agents, and Effects Sylvie Yaacoub
    Eastern Michigan University DigitalCommons@EMU Senior Honors Theses Honors College 2015 General Anesthesia: A Literature Review - Induction, Mechanism, Agents, and Effects Sylvie Yaacoub Follow this and additional works at: http://commons.emich.edu/honors Recommended Citation Yaacoub, Sylvie, "General Anesthesia: A Literature Review - Induction, Mechanism, Agents, and Effects" (2015). Senior Honors Theses. 425. http://commons.emich.edu/honors/425 This Open Access Senior Honors Thesis is brought to you for free and open access by the Honors College at DigitalCommons@EMU. It has been accepted for inclusion in Senior Honors Theses by an authorized administrator of DigitalCommons@EMU. For more information, please contact lib- [email protected]. General Anesthesia: A Literature Review - Induction, Mechanism, Agents, and Effects Degree Type Open Access Senior Honors Thesis Department Nursing First Advisor Sandra Hines Keywords General Anesthesia, General Anesthetics, Effects on Cancer, Developmental Deficits in Pediatric Population, Cancer Proognosis, Consciousness This open access senior honors thesis is available at DigitalCommons@EMU: http://commons.emich.edu/honors/425 General Anesthesia A Literature Review Induction, Mechanism, Agents, and Effects by Sylvie Y aacoub A Senior Thesis Submitted to the Eastern Michigan University Honors College in Partial Fulfillment for the Requirements for Graduation with Honors in Nursing Approved in Ypsilanti, MI on this date ¥ ~ ,L I> 1-;; General Anesthesia A Literature Review Induction, Mechanism, Agents,
    [Show full text]