Gottfried Werner

The incorporation of putrescine carbon into γ-aminobutyric acid in rat liver and brain in vivo

Summary 14C from intraperitoneally injected [1,4-14C]putrescine was incorporated into γ-aminobutyric acid of liver and brain, along with its incorporation into the polyamines spermidine and spermine and into glutamic acid. The results obtained from liver can be interpreted as follows. Formation of peripheral γ-aminobutyric acid occurs to a considerable extent on a pathway not including glutamic acid decarboxylation. γ-Aminobutyric acid supposedly plays a role as intermediary substance in the oxidation of putrescine to CO2 and of substances which are transformed into putrescine; especially in organs in which this amino acid is not connected with the functionality of inhibitory neurones. Brain γ-aminobutyric acid exhibited a lower specific activity than glutamic acid. Therefore, the present data do not prove the existence of a pathway of γ-aminobutyric acid formation parallel with the glutamic acid decarboxylation in brain.

Interrelationships between polyamines and nucleic acids. III. Metabolic and autoradiographic studies of putrescine in mouse brain

Abstract To corroborate further the supposed functional relation between polyamines and nucleic acids, regional differences of the metabolism of putrescine in vivo have been studied with an autoradiographic technique. The regions of maximal synthesis of spermidine and spermine from radioactive putrescine were identical to a large extent with those of maximal protein and RNA synthesis. As had been already demonstrated for radioactive amino acids and for some precursors of RNA, we also found the highest accumulations of radioacttivity from intraperitoneally injected [2,3- 3 H]-putrescine to be generally in the region rich in nerve cells. The highest proportion of radioactivity was represented by spermidine and spermine. In the case of putrescine some tracts contained even more radioactive material than the surrounding gray matter, and these tracts were not labelled after the application of radioactive amino acids. An appreciable amount of radioactive putrescine in mouse brain was observed even 28 days after the administration of the radioactive compound. This suggests the existence of a degradative pathway from spermidine and spermine to putrescine in the mammalian organism.

Detection of δ9-tetrahydrocannabinol in saliva of men by means of thin-layer chromatography and mass spectrometry

Abstract The saliva of men is suggested as a useful biological material for revealing cannabis abuse. Ten subject volunteered and smoked a single tobacco cigarette that contained an amount of cannabis equivalent to 2.8 mg of δ 9 -tetrahydrocannabinol. After smoking, the δ 9 -tetrahydrocannabinol was detected in the saliva extracts by means of two-dimensional thin-layer chromatography and mass spectrometry for periods up to 2 h.

Isoelectric focusing in continuous-flow electrophoresis: I. Separation of mixed red blood cells of different species

Abstract A noncommercial continuous-flow isoelectric focusing (CIF) apparatus which was formerly applied to separate mixtures of proteins was used to study the separation of red blood cells (RBCs) of different species. A mixture of human, mouse and rabbit erythrocytes, a good model for demonstration of cell separation by CIF, was completely separated into the three components. The separation was performed by isoelectric focusing in pH gradients, 3–10 and 5–7, using Ampholine carrier ampholytes at a field strength of 110 V/cm and a flow-through time of RBCs of 7 min. The isoionic points of human RBCs both determined by CIF and calculated from electrophoretic mobility measurements by extrapolation to zero electrophoretic mobility and zero ionic strength were found at pH 5.6–5.7. The method of CIF which is presently used to isolate a pure lysosomal fraction seems to be a valuable method for the separation of mixtures of cells or cell organelles.

Bestimmung von ?1 und ?1(6)-tetrahydrocannabinol in Blut, Urin und Speichel von Haschisch-Rauchern

Das aus d e m K u r v e n v e r l a u f n a c h [5] zu e rmi t t e lnde t s betr~gt e twa 7,5 h, tG, e twa 3 h, t M e twa 2 h. N a c h d e m ersten M a x i m u m verl/~uft die K u r v e Iiir ein l~ngeres In te rva l l nahe der Abszisse, bevor sie erl leut ans te ig t . Aus der zei t l ichen Differenz der be iden M a x i m a ergibt sich ein I c yon e twa 33 h ulld d a m i t ein tG~ rol l e twa 20 h. Dieser Ver lauf der t s be legt Mndeut ig die F~higke i t der H e p a t o c y t e n , nach Teilh e p a t e k t o m i e n i ch t n u r e inmalig, sonde rn wiederholt in den Zellzyklus einzutreten, wobei tG~ des zwei ten Zyklus gegeni iber IG: des e rs ten Zellzyklus, d e m In te rva l I zwischell Te i lhepa t ek tomie u n d Beg inn der D N S S y n t h e s e , n i ch t v e r k f r z t ist. Obwohl die Hepa tocy te l l berei ts einell Zel lzyklus nach der Leber resek t ion du rch l au fen haben , s ind fiir den Beg inn der zweitell S -Phase of fenbar eine e rneu te I n d u k t i o n a n d ~hlllich kompl iz ier te p r epa ra t i ve S y n t h e s e n erforderl ich wie in G 1 des ersten Zyklus .

Eine Methode zur autoradiographischen Darstellung hydrophiler Substanzen in biologischem Material

Mit der beschriebenen autoradiographischen Methode lassen sich auch niedermolekulare hydrophile Substanzen, die mit Radioisotopen markiert sind, darstellen. Einige Beispiele zeigen die Leistungsfähigkeit des Verfahrens.

Ein Beitrag zur Technik der Autoradiographie elektronenmikroskopischer Präparate

SummaryA practical method for electronmicroscopic autoradiography of plastic embedded sections is presented. The method is especially useful for making a large number of autoradiographs of serial sections effeciently. An apparatus automating photographic development, fixing, and contrast staining is described.ZusammenfassungEin Verfahren zur Autoradiographie von Ultradünnschnitten plastikeingebetteter elektronenmikroskopischer Präparate wird ausführlich beschrieben. Durch den Einsatz eines Automaten für die photographische Entwicklung und Nachkontrastierung der Präparate gelingt es ohne größeren zeitlichen oder personellen Aufwand unter reproduzierbaren Bedingungen rationell zu arbeiten. Da bei der beschriebenen Methodik praktisch kein Präparatverlust auftritt, eignet sich das Verfahren besonders auch für Untersuchungen an Schnittserien.

Metabolism and autoradiographic distribution of Δ8- and Δ9 tetrahydrocannabinol in some organs of the monkey callithrix jacchus

SummaryMetabolism and autoradiographic distribution of the two isomeric tetrahydrocannabinols, (2,4-14C)-Δ8-THC and (2,4-14C)-Δ9-THC), were studied in the marmoset Callithrix jacchus. Of the two cannabinoids, Δ8-THC had a slower initial rate of biotransformation to the psychopharmacologically more potent 11-hydroxylated metabolite. This may explain the minor psychopharmacological activity of the Δ8-isomer. In glandular tissues an accumulation of unchanged Δ9-THC was observed.Autoradiography revealed characteristic label distributions in some organs 30 min after the administration of the drugs. This labelling pattern was found to be changed after a 6-hr incorporation period. The autoradiographic distribution of Δ8- and Δ9-THC appeared to be identical.

Elektrophorese im trägerfreien Pufferstrom

A separation chamber for continuous electrophoresis in a free flowing buffer curtain is described. The feature of our construction is the non-planar separation chamber and the assemblage of the electrodes within a constructive unit, the electrode block. This effects a handy and mechanically stable device. Furthermore, it enables the set-up of separation chambers of different size and shape, simply by the exchange of the chamber units.ZusammenfassungEs wird die Konstruktion einer Trennkammer für die kontinuierliche Ablenkungselektrophorese im trägerfreien Pufferstrom mitgeteilt. Das besondere Kennzeichen unseres Gerätes ist die nicht ebene Trennkammer und die konstruktive Zusammenfassung der Elektroden in einem Block. Durch diese Bauweise wird eine handliche, mechanisch stabile Trennkammer großer Präzision gewährleistet. Darüber hinaus kann durch den Austausch der Kammerteile der Aufbau von Trennkammern verschiedener Dimensionen und Formen bewirkt werden.

Beziehungen zwischen Polyaminen und Nucleinsäuren

SummaryExposition of freeze-dried cryostat sections of tissue to formaldehyde vapour caused the binding of the polyamines spermine and spermidine, so that they were not extractable with acids. In persecution of this observation we isolated the protein, nucleic acid and lipid fraction of liver after the injection of radioactive putrescine to mice, at a time when radioactivity was mostly accumulated in spermidine. Though in model experiments the binding of the amines was most extensive to protein, we observed the most pronounced binding of radioactive spermidine to the nucleic acid fraction of the liver. This indicates, that spermidine is preferentially localised in vivo in nucleic acid containing structures.ZusammenfassungAusgehend von der Beobachtung, daß sich die Extrahierbarkeit von Putrescin und seiner Metabolite nach dem Exponieren von Kryostatschnitten in der sich bei 80° C über Paraformaldehyd ausbildenden Atmosphäre wesentlich verringert, wurde der Versuch unternommen, die unter Formaldehyd-Einfluß eintretende Fixierung der Polyamine zum Studium der intrazellulären Verteilung speziell von Putrescin und Spermidin zu nutzen. Die Fraktionierung von Lebergewebe in Lipide, Proteine und Nucleinsäuren ergab eine erhebliche Anreicherung von Spermidin in der Nucleinsäurenfraktion. Unter Berücksichtigung der Ergebnisse von Modellversuchen sind unsere Beobachtungen als ein Hinweis für die bevorzugte Lokalisation von Spermidin in nucleinsäurenhaltigen Strukturen der intakten Zelle zu deuten.