4.3 M ETHODEN
4.3.16 Darstellung und statistische Bewertung der Messwerte
Standardmäßig wurden für einen Messwert je vier Einzelwerte auf einer 96-Loch-Platte gemittelt. Aus den so erhaltenen Messwerten aus n unabhängigen Versuchen wurden dann Mittelwerte MW und die Standardabweichung s berechnet. Auf signifikante Unterschiede wurden mit dem Wilcoxon-Test geprüft (Signifikanzniveau p < 0,05). Bei Abbildungen von Thrombinbildungskurven wurde auf das Einzeichnen der Standardabweichungen um die Mittelwerte der einzelnen Messpunkte aus Gründen der Übersichtlichkeit verzichtet.
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6 Anhang
6.1 Verzeichnis der im Text verwendeten Abkürzungen
α2-Makroglobulin a2M
Abu L-alpha-Aminobuttersäure Ac Acetyl
AMC Aminomethylcumarin antiTF-AK anti-Tissue Factor-Antikörper
APAAP-Komplex alkalische Phosphatase und monoklonaler anti-alkalischer Phosphatase-Komplex
AUC Area under the curve, Fläche unter der Thrombinbildungskurve C1-INH C1-Esterase-Inhibitor
ELISA enzyme linked immuno sorbent assay FACS fluorescence-activated cell sorting FKS fötales Kälberserum
FVIIai inaktivierter FVIIa
GGACK 1,5 Dansyl-Glu-Gly-Arg Chlormethylketon HHT Hexahydrotyrosin
HSA humanes Serumalbumin
HUVEC human umbilical vein endothelial cells (Endothelzellen aus Nabelschnurvenen)
IU international unit (Kennzeichnung der Enzymaktivität) kD Kilodalton
kcat kinetische Konstante
Km Michaelis-Konstante
LPS bakterielle Lipopolysaccharide
Me Methyl
MG Molekulargewicht
MOC Methoxycarbonyl MP Mikropartikel MW Mittelwert
PAF platelet activating factor
PC Phosphatidylcholin Pip Piperidyl
PL Phospholipide pNA para-Nitroanilin POD Peroxidase
PPP platelet poor plasma (plättchenarmes Plasma) PRP platelet rich plasma (plättchenreiches Plasma) PS Phosphatidylserin
s Standardabweichung SMC smooth muscle cells (glatte Muskelzellen)
sTF soluble Tissue Factor, extrazelluläre Tissue Factor-Domäne TF Tissue Factor (Gewebefaktor)
TFPI Tissue Factor Pathway Inhibitor TM Thrombomodulin TNFα Tumor Nekrose Faktor α
In der Literatur allgemein übliche Abkürzungen finden hier keine Erwähnung.
6.2 Veröffentlichungen
Teile dieser Arbeit wurden veröffentlicht:
Fischer A, Prasa D, Heller R, Stürzebecher J. Determination of procoagulant activity of human umbilical vein endothelial cells by continuous measurement of thrombin generation. Annals of Hematol 78, Suppl. Abstracts (1999)
Fischer A, Prasa D, Heller R, Stürzebecher J. Continuous measurement of thrombin generation-a method to determine procoagulant activity of human umbilical vein endothelial cells. Thromb Haemost 82, Suppl. Abstracts (1999)
Bretschneider E, Prasa D, Fischer A, Stürzebecher J. Human vascular smooth muscle cells can initiate the activation of the extrinsic coagulation pathway. Annals of Hematol 78, Suppl. Abstracts (1999)
Bretschneider E, Wittpoth M, Fischer A, Prasa D, Stürzebecher J, Braun M, Glusa E, Schrör K. Human vascular smooth muscle cells generate thrombin via the activation of the extrinsic coagulation pathway. Thromb Haemost 82, Suppl. Abstracts (1999) Bretschneider E, Braun M, Fischer A, Wittpoth M, Glusa E, Schrör K. Factor Xa acts as a
PDGF-independent mitogen in human vascular smooth muscle cells. Thromb Haemost 84: 499-505 (2000)
ERKLÄRUNG
Ich versichere, dass ich meine Dissertation
„Kontinuierliche Messung der zellgetriggerten Thrombinbildung Entwicklung einer Methode
zur Bestimmung der prokoagulatorischen Aktivität von Zellen“
selbständig ohne unerlaubte Hilfe angefertigt und mich dabei keiner anderen als der von mir ausdrücklich bezeichneten Quellen bedient habe.
Die Dissertation wurde in der jetzigen oder einer ähnlichen Form noch bei keiner anderen Hochschule eingereicht und hat noch keinen sonstigen Prüfungszwecken gedient.
Marburg, den 16. Mai 2003
DANKSAGUNG
Die vorliegende Arbeit entstand am Zentrum für Vaskuläre Biologie und Medizin/
Arbeitsgruppe „Biochemie der Hämostase“ der Friedrich-Schiller-Universität Jena unter Leitung von Dozent Dr. Jörg Stürzebecher in Zusammenarbeit mit dem Institut für Pharmazeutische Chemie/ Arbeitskreis Professor Dr. Gerhard Klebe der Universität Marburg.
Für die mir in zahlreichen Diskussionen gebotenen Anregungen und die stets sehr freundliche und großzügige Unterstützung möchte ich Herrn Dr. Stürzebecher herzlich danken. In meinen Dank möchte ich insbesondere Frau Dr. Dagmar Prasa einschließen, die immer zum kritischen Meinungsaustausch bereit war.
Frau Dr. Heller (Friedrich-Schiller-Universität Jena) und ihren Mitarbeitern danke ich für die Einführung in Techniken der Zellkultur, die Bereitstellung von biologischem Material und die Hilfestellung bei der Präparation von TF-mRNA. Ebenso gilt mein Dank Herrn Dr.
Gräbner (Friedrich-Schiller-Universität Jena) für die Unterweisung in immunhisto-chemischen Techniken. Frau Dr. Bretschneider (Friedrich-Schiller-Universität Jena) danke ich für die Bereitstellung glatter Gefäßmuskelzellen. Dem Arbeitskreis um Herrn Dr.
Luther und Frau Dr. Albrecht (Technische Universität Dresden) danke ich für die Gabe von TF-Antikörpern und von Materialien für den TF-ELISA sowie für die gemeinsamen Messungen an verschiedenen Zelllinien.