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Saccharomyces cerevisiae possesses two glycogenin isoforms (designated as Glg1p and Glg2p) that both contain a conserved tyrosine residue, Tyr232. However, Glg2p possesses an additional tyrosine residue, Tyr230 and therefore two potential autoglucosylation sites. Glucosylation of Glg2p was studied using both matrix-assisted laser desorption ionization and electrospray quadrupole time of flight mass spectrometry. Glg2p, carrying a C-terminal (His(6)) tag, was produced in Escherichia coli and purified. By tryptic digestion and reversed phase chromatography a peptide (residues 219-246 of the complete Glg2p sequence) was isolated that contained 4-25 glucosyl residues. Following incubation of Glg2p with UDPglucose, more than 36 glucosyl residues were covalently bound to this peptide. Using a combination of cyanogen bromide cleavage of the protein backbone, enzymatic hydrolysis of glycosidic bonds and reversed phase chromatography, mono- and diglucosylated peptides having the sequence PNYGYQSSPAM were generated. MS/MS spectra revealed that glucosyl residues were attached to both Tyr232 and Tyr230 within the same peptide. The formation of the highly glucosylated eukaryotic Glg2p did not favour the bacterial glycogen accumulation. Under various experimental conditions Glg2p-producing cells accumulated approximately 30% less glycogen than a control transformed with a Glg2p lacking plasmid. The size distribution of the glycogen and extractable activities of several glycogen-related enzymes were essentially unchanged. As revealed by high performance anion exchange chromatography, the intracellular maltooligosaccharide pattern of the bacterial cells expressing the functional eukaryotic transgene was significantly altered. Thus, the eukaryotic glycogenin appears to be incompatible with the bacterial initiation of glycogen biosynthesis
SIRT6 is a NAD(+)-dependent deacetylase that modulates chromatin structure and safeguards genomic stability. Until now, SIRT6 has been assigned to the nucleus and only nuclear targets of SIRT6 are known. Here, we demonstrate that in response to stress, C. elegans SIR-2.4 and its mammalian orthologue SIRT6 localize to cytoplasmic stress granules, interact with various stress granule components and induce their assembly. Loss of SIRT6 or inhibition of its catalytic activity in mouse embryonic fibroblasts impairs stress granule formation and delays disassembly during recovery, whereas deficiency of SIR-2.4 diminishes maintenance of P granules and decreases survival of C. elegans under stress conditions. Our findings uncover a novel, evolutionary conserved function of SIRT6 in the maintenance of stress granules in response to stress.
Spurensuche Am Neuen Palais
(2016)
Sechs kurze Krimis über abgründige Verbrechen, unverwechselbare Tatorte (Potsdam-Sanssouci u. a.) und höchst eigenwillige Ermittler/innen, die ein hohes Risiko eingehen, um die rätselhaften Fälle zu lösen und schließlich für Klarheit und Aufklärung zu sorgen.
Unterhaltsam und spannend erzählt von Stefanie Börnicke & Friederike Weimar, Paula-Sophie Brink, Steven Dewart, Martin Thormann, Simone Weilandt. Mit einem Nachwort von Brunhilde Wehinger.
Das 12. Herbsttreffen Patholinguistik mit dem Schwerpunktthema »Weg(e) mit dem Stottern: Therapie und Selbsthilfe für Kinder und Erwachsene« fand am 24.11.2018 in Potsdam statt. Das Herbsttreffen wird seit 2007 jährlich vom Verband für Patholinguistik e.V. (vpl) durchgeführt. Der vorliegende Tagungsband beinhaltet die Vorträge zum Schwerpunktthema sowie Beiträge der Posterpräsentationen zu weiteren Themen aus der sprachtherapeutischen Forschung und Praxis.
Oligosaccharides composed of 2-acetamido-2-deoxy-D-glucopyranose (GlcNAc) and/or 2-amino-2-deoxy-D- glucopyranose (GlcN) were prepd. by chem. degrdn. of chitin or chitosan and sepd. by gel permeation chromatog. Oligosaccharides obtained after enzymic hydrolysis of chitosan [FA 0.19] with a fungal chitinase were derivatized by reductive amination with 2-aminoacridone and sequenced by matrix-assisted laser desorption ionization time-of-flight postsource decay (PSD) mass spectrometry (MS). The sequence of a trimer, D1A2, was established as D-A-A. The compn. of a hexamer D3A3 was .apprx.65% D-A-D-D-A-A and 35% D-D-A-D-A-A. The PSD MS of a nonamer D5A4-amac revealed four isobaric species D-X-Y-D-X-Y-D-A-A, where A is GlcNAc, D is GlcN, and X and Y (X ¹ Y) are mutually either D or A. This structure motif was also obsd. in a dodecamer D7A5 which was composed of eight isobaric sequences of the general formula (D-X-Y)3- D-A-A.
Repetitive overhead movements have been identified as a main risk factor to develop shoulder complaints with scapular muscle activity being altered. Reliable assessment of muscle activity is essential to differentiate between symptomatic and asymptomatic individuals. Therefore, the present study aimed to investigate the intra-and inter-session reliability of scapular muscle activity during maximal isokinetic shoulder flexion and extension. Eleven asymptomatic adults performed maximum effort isokinetic shoulder flexion and extension (concentric and eccentric at 60 degrees/s) in a test-retest design. Muscle activity of the upper and lower trapezius and serratus anterior was assessed by sEMG. Root Mean Square was calculated for whole ROM and single movement phases of absolute and normalized muscle activity. Absolute (Bland-Altman analysis (Bias, LoA), Minimal detectable change (MDC)) and relative reliability parameters (Intraclass correlation coefficient (ICC), coefficient of variation (CV)/test-retest variability (TRV)) were utilized for the evaluation of reproducibility. Intra-session reliability revealed ICCs between 0.56 and 0.98, averaged CVs of 18% and average MDCs of 81 mV. Inter-session reliability resulted in ICCs between 0.13 and 0.93, averaged TRVs of 21%, average MDCs of 15% and systematic and random error between -8 +/- 60% and 12 +/- 36%. Scapular muscle activity assessed in overhead movements can be measured reliably under maximum load conditions, though variability is dependent on the movement phase. Measurement variability does not exceed magnitudes of altered scapular muscle activities as reported in previous studies. Therefore, maximum load application is a promising approach for the evaluation of changes in scapular control related to pathologies. (C) 2017 Elsevier Ltd. All rights reserved.
Heterochitooligosaccharides possess interesting biol. properties. Isobaric mixts. of such linear heterochitooligosaccharides can be obtained by chem. or enzymic degrdn. of chitosan. However, the sepn. of such mixts. is a challenging anal. problem which is so far unresolved. It is shown that these isobaric mixts. can be sequenced and quantified simultaneously using std. derivatization and multistage tandem mass spectrometric techniques. A linear ion trap mass spectrometer equipped with a vacuum matrix-assisted laser desorption ionization (vMALDI) source is used to perform MS2 as well as MS3 expts. [on SciFinder (R)].
Portal Wissen = Zeit
(2014)
„Was ist also 'Zeit'?“ seufzt Augustinus von Hippo im 11. Buch seiner „Confessiones“ melancholisch, und fährt fort „Wenn mich niemand danach fragt, weiß ich es; will ich einem Fragenden es erklären, weiß ich es nicht.“ Auch heute, 1584 Jahre nach Augustinus, erscheint 'Zeit' immer noch rätselhaft. Abhandlungen über das Wesen der Zeit füllen Bibliotheken. Oder eben dieses Heft.
Wesensfragen sind den modernen Wissenschaften allerdings fremd. Zeit ist – zumindest in der Physik – unproblematisch. „Time is defined so that Motion looks simple“ erkärt man kurz und trocken, und verabschiedet sich damit vom Augustinischen Rätsel oder der Newtonschen Vorstellung einer absoluten Zeit, deren mathematischen Fluss man durch irdische Instrumente eh immer nur näherungsweise erfassen kann.
In der Alltagssprache, selbst in den Wissenschaften, reden wir zwar weiterhin vom Fluss der Zeit, aber Zeit ist schon lange keine natürliche Gegebenheit mehr. Zeit ist vielmehr ein konventioneller Ordnungsparameter für Änderung und Bewegung. Geordnet werden Prozesse, indem eine Klasse von Prozessen als Zählsystem dient, um andere Prozesse mit ihnen zu vergleichen und anhand der temporären Kategorien „vorher“, „während“ und „nachher“ anzuordnen.
Zu Galileis Zeiten galt der eigene Pulsschlag als Zeitstandard für den Flug von Kanonenkugeln. Mit zunehmender Verfeinerung der Untersuchungsmethoden erschien das zu unpraktisch: Die Weg-Zeit-Diagramme frei fliegender Kanonenkugeln erweisen sich in diesem Standard ziemlich verwackelt, schlecht reproduzierbar, und keineswegs „simpel“. Heutzutage greift man zu Cäsium-Atomen. Demnach dauert ein Prozess eine Sekunde, wenn ein 133Cs-Atom genau 9 192 631 770 Schwingungen zwischen zwei sogenannten Hyperfeinzuständen des Grundzustands vollführt hat. Und ein Meter ist die Entfernung, die Licht im Vakuum in exakt 1/299 792 458 Sekunden zurücklegt. Glücklicherweise sind diese Daten im General Positioning System GPS hart kodiert, so dass der Nutzer sie nicht jedes Mal aufs Neue eingeben muss, wenn er wissen will, wo er ist. Aber schon morgen muss er sich vielleicht ein Applet runterladen, weil der Zeitstandard durch raffinierte Übergänge in Ytterbium ersetzt wurde.
Der konventionelle Charakter des Zeitbegriffs sollte nicht dazu verführen zu glauben, alles sei irgendwie relativ und daher willkürlich. Die Beziehung eines Pulsschlags zu einer Atomuhr ist absolut, und genauso real, wie die Beziehung einer Sanduhr zum Lauf der Sonne. Die exakten Wissenschaften sind Beziehungswissenschaften. Sie handeln nicht vom Ding an sich, was Newton und Kant noch geträumt haben, sondern von Beziehungen – worauf schon Leibniz und später Mach hingewiesen haben.
Kein Wunder, dass sich für andere Wissenschaften der physikalische Zeit-Standard als ziemlich unpraktisch erweist. Der Psychologie der Zeitwahrnehmung entnehmen wir – und jeder wird das bestätigen können – dass das gefühlte Alter durchaus verschieden ist vom physikalischen Alter. Je älter man ist, desto kürzer erscheinen einem die Jahre.
Unter der einfachen Annahme, dass die gefühlte Dauer umgekehrt proportional zum physikalischen Alter ist, und man als Zwanzigjähriger ein physikalisches Jahr auch psychologisch als ein Jahr empfindet, ergibt sich der erstaunliche Befund, dass man mit 90 Jahren 90 Jahre ist. Und – bei einer angenommenen Lebenserwartung von 90 Jahren – mit 20 (bzw. 40) physikalischen Jahren bereits 67 (bzw. 82) Prozent seiner gefühlten Lebenszeit hinter sich hat.
Bevor man angesichts der „Relativität von Zeit“ selbst in Melancholie versinkt, vielleicht die Fortsetzung des Eingangszitats von Augustinus: „Aber zuversichtlich behaupte ich zu wissen, dass es vergangene Zeit nicht gäbe, wenn nichts verginge, und nicht künftige Zeit, wenn nichts herankäme, und nicht gegenwärtige Zeit wenn nichts seiend wäre.“ Tja – oder mit Bob Dylan „The times they're a changing“.
Ich wünsche Ihnen eine spannende Zeit bei der Lektüre dieser Ausgabe.
Prof. Dr. Martin Wilkens
Professor für Quantenoptik
Portal Wissen = Time
(2014)
“What then is time?”, Augustine of Hippo sighs melancholically in Book XI of “Confessions” and continues, “If no one asks me, I know; if I want to explain it to a questioner, I don’t know.” Even today, 1584 years after Augustine, time still appears mysterious. Treatises about the essence of time fill whole libraries – and this magazine.
However, questions of essence are alien to modern sciences. Time is – at least in physics – unproblematic: “Time is defined so that motion looks simple”, briefly and prosaically phrased, waves goodbye to Augustine’s riddle and to the Newtonian concept of absolute time, whose mathematical flow can only be approximately recorded with earthly instruments anyway.
In our everyday language and even in science we still speak of the flow of time but time has not been a natural condition for quite a while now. It is rather a conventional order parameter for change and movement. Processes are arranged by using a class of processes as a counting system in order to compare other processes and to organize them with the help of the temporary categories “before”, “during”, and “after”.
During Galileo’s time one’s own pulse was seen as the time standard for the flight of cannon balls. More sophisticated examination methods later made this seem too impractical. The distance-time diagrams of free-flying cannon balls turned out to be rather imprecise, difficult to replicate, and in no way “simple”. Nowadays, we use cesium atoms. A process is said to take one second when a caesium-133 atom completes 9,192,631,770 periods of the radiation corresponding to the transition between two hyperfine levels of the ground state. A meter is the length of the path travelled by light in a vacuum in exactly 1/299,792,458 of a second. Fortunately, these data are hard-coded in the Global Positioning System GPS so users do not have to reenter them each time they want to know where they are. In the future, however, they might have to download an app because the time standard has been replaced by sophisticated transitions to ytterbium.
The conventional character of the time concept should not tempt us to believe that everything is somehow relative and, as a result, arbitrary. The relation of one’s own pulse to an atomic clock is absolute and as real as the relation of an hourglass to the path of the sun. The exact sciences are relational sciences. They are not about the thing-initself as Newton and Kant dreamt, but rather about relations as Leibniz and, later, Mach pointed out.
It is not surprising that the physical time standard turned out to be rather impractical for other scientists. The psychology of time perception tells us – and you will all agree – that the perceived age is quite different from the physical age. The older we get the shorter the years seem. If we simply assume that perceived duration is inversely related to physical age and that a 20-year old also perceives a physical year as a psychological one, we come to the surprising discovery that at 90 years we are 90 years old. With an assumed life expectancy of 90 years, 67% (or 82%) of your felt lifetime is behind you at the age of 20 (or 40) physical years.
Before we start to wallow in melancholy in the face of the “relativity of time”, let me again quote Augustine. “But at any rate this much I dare affirm I know: that if nothing passed there would be no past time; if nothing were approaching, there would be no future time; if nothing were, there would be no present time.” Well, – or as Bob Dylan sings “The times they are a-changin”.
I wish you an exciting time reading this issue.
Prof. Martin Wilkens
Professor of Quantum Optics