Die vorliegende Arbeit untersucht in einer Population von 132 gesunden Probanden die Hämostase mittels Calibrated Automated Thrombogram (CAT) und Rotationsthromboelastometrie (ROTEM). CAT wurde im plätchenarmen Plasma mit einer tissue factor (TF) von 1 und 5 pM durchgeführt. Lag time, Thrombin peak, Time to thrombin peak und das endogene Thrombin Potential (ETP) wurden ermittelt. ROTEM wurde ohne Aktivator durchgeführt (NATEM) und die Daten für Gerinnungszeit (clotting time, CT), Gerinnselbildungszeit, Alpha Winkel und maximale Gerinnselfestigkeit (MCF) mit den Daten der Thrombingenerierung korreliert. Es zeigte sich eine positive aber nicht lineare Korrelation bezüglich Alter versus lag time und time to peak, sowie eine annähernd lineare Korrelation bezüglich Alter versus thrombin peak und ETP. Für ROTEM konnte eine positive Korrelation bezüglich Alter versus MCF und Alpha Winkel, aber eine negative Korrelation bezüglich Alter versus CT dargestellt werden. In der Gegenüberstellung beider Assays korrelierten Thrombin peak und ETP (aktiviert mit einer TF Konzentration von 5 pM) signifikant mit dem Alpha Winkel und der MCF. Alle signifikanten Korrelationen zeigten lediglich eine moderate Regressionssteigung. / Published data on thrombin generation variables and their correlation with thromboelastometry in the healthy population are scarce. This study aimed at assessing thrombin generation in adults and its correlation to classical rotational thromboelastometry (ROTEM). Methods: Thrombin generation was measured in platelet-poor plasma from healthy volunteers using the calibrated automated thrombogram (CAT) with 1 and 5 pmol/l tissue factor final concentration. Lag time, thrombin peak, time to thrombin peak and endogenous thrombin potential (ETP) were analyzed. ROTEM was performed without activator (NATEM) and data for clotting time, alpha angle, clot formation time and maximum clot firmness were correlated with those of thrombin generation. Results: Altogether 132 persons (72 men, 60 women; median age: 48.0 years) were included. There was a positive non-linear correlation for age versus lag time (p < 0.001) and time to peak (p = 0.001), and almost linear correlation for age versus thrombin peak (p = 0.024) and ETP (p = 0.001), although with a moderate regression slope. Regarding ROTEM, there was a positive correlation between age and maximum clot firmness and alpha angle (p = 0.001), but a negative correlation between age and clotting time (p = 0.039). Comparing both assays, thrombin peak and ETP measured with a final tissue factor concentration of 5 pmol/l correlated significantly with alpha angle and maximum clot firmness. Conclusion: The age-related changes in CAT and ROTEM variables among adults are not linear. There is a significant correlation, although with a moderate slope, between data from CAT measured with 5 pmol/l tissue factor and ROTEM.

Die vorliegende Arbeit untersucht in einer Population von 132 gesunden Probanden die Hämostase mittels Calibrated Automated Thrombogram (CAT) und Rotationsthromboelastometrie (ROTEM). CAT wurde im plätchenarmen Plasma mit einer tissue factor (TF) von 1 und 5 pM durchgeführt. Lag time, Thrombin peak, Time to thrombin peak und das endogene Thrombin Potential (ETP) wurden ermittelt. ROTEM wurde ohne Aktivator durchgeführt (NATEM) und die Daten für Gerinnungszeit (clotting time, CT), Gerinnselbildungszeit, Alpha Winkel und maximale Gerinnselfestigkeit (MCF) mit den Daten der Thrombingenerierung korreliert. Es zeigte sich eine positive aber nicht lineare Korrelation bezüglich Alter versus lag time und time to peak, sowie eine annähernd lineare Korrelation bezüglich Alter versus thrombin peak und ETP. Für ROTEM konnte eine positive Korrelation bezüglich Alter versus MCF und Alpha Winkel, aber eine negative Korrelation bezüglich Alter versus CT dargestellt werden. In der Gegenüberstellung beider Assays korrelierten Thrombin peak und ETP (aktiviert mit einer TF Konzentration von 5 pM) signifikant mit dem Alpha Winkel und der MCF. Alle signifikanten Korrelationen zeigten lediglich eine moderate Regressionssteigung. / Published data on thrombin generation variables and their correlation with thromboelastometry in the healthy population are scarce. This study aimed at assessing thrombin generation in adults and its correlation to classical rotational thromboelastometry (ROTEM). Methods: Thrombin generation was measured in platelet-poor plasma from healthy volunteers using the calibrated automated thrombogram (CAT) with 1 and 5 pmol/l tissue factor final concentration. Lag time, thrombin peak, time to thrombin peak and endogenous thrombin potential (ETP) were analyzed. ROTEM was performed without activator (NATEM) and data for clotting time, alpha angle, clot formation time and maximum clot firmness were correlated with those of thrombin generation. Results: Altogether 132 persons (72 men, 60 women; median age: 48.0 years) were included. There was a positive non-linear correlation for age versus lag time (p < 0.001) and time to peak (p = 0.001), and almost linear correlation for age versus thrombin peak (p = 0.024) and ETP (p = 0.001), although with a moderate regression slope. Regarding ROTEM, there was a positive correlation between age and maximum clot firmness and alpha angle (p = 0.001), but a negative correlation between age and clotting time (p = 0.039). Comparing both assays, thrombin peak and ETP measured with a final tissue factor concentration of 5 pmol/l correlated significantly with alpha angle and maximum clot firmness. Conclusion: The age-related changes in CAT and ROTEM variables among adults are not linear. There is a significant correlation, although with a moderate slope, between data from CAT measured with 5 pmol/l tissue factor and ROTEM.

Les tests de coagulation actuellement disponibles en médecine vétérinaire ne parviennent pas à détecter l’état hypercoagulable de manière fiable. Les tests usuels de l’hémostase (ex : PT, PTT, antithrombine, D-dimères, etc.) n’évaluent malheureusement qu’une infime portion de la coagulation sanguine. Les tests globaux de l’hémostase, incluant la thromboélastographie et la génération de thrombine, permettent une évaluation plus complète de l’hémostase, incluant l’équilibre entre les facteurs procoagulants et anticoagulants. Ceux-ci comportent cependant encore certaines lacunes quant à leur performance diagnostique. L’étude présentée dans ce mémoire propose des modifications à la génération de thrombine conventionnelle dans le but d’identifier un état hypercoagulable chez des chiens malades. Plus précisément, nous avons comparé les effets in vitro de la thrombomoduline et de daltéparine sur la génération de thrombine réalisée chez des chiens sains et des chiens malades, pour démontrer une déficience en anticoagulants endogènes (i.e. protéine C et antithrombine) chez ces derniers. Pour se faire, 10 chiens sains et 10 chiens malades ont été sélectionnés en fonction de leur historique, examen physique et analyses sanguines. Du plasma pauvre en plaquettes a été obtenu pour chaque chien à partir de sang total citraté, aliquoté et congelé à -80 ̊Celsius pendant moins de 18 mois. La génération de thrombine a été mesurée sur le plasma décongelé à l'aide de la méthode Calibrated Automated Thrombogram. Les mesures ont d’abord été réalisées sans anticoagulant, puis avec l’addition de thrombomoduline et de trois concentrations de daltéparine (0,2, 0,4 et 0,6 U/ml). Le potentiel de thrombine endogène (ETP), le temps d’initiation (Lag time), le temps pour l’atteinte du pic (TTPeak) et le pic (Peak) ont été obtenus et analysés à l'aide d'un modèle linéaire mixte (p < 0,05). Aucune différence n’était observée entre les groupes avant l’ajout d’anticoagulants. L’ajout de thrombomoduline et de daltéparine entraînait une diminution significative de l'ETP et du Peak dans les deux groupes. Cependant, l’effet de ces deux anticoagulants étaient significativement moins marqué dans le groupe malade par rapport au groupe sain, à l'exception du Peak aux concentrations de daltéparine les plus élevées (0,4 U/ml et 0,6 U/ml). Cette modification de la génération de thrombine conventionnelle permet donc de mettre en évidence un déficit en anticoagulant endogène chez les chiens malades, chez lesquels la génération de thrombine est moins affectée par l’ajout in vitro de substances anticoagulantes. Par conséquent, effectuer la génération de thrombine avec et sans l'ajout d'anticoagulants exogènes pourrait être bénéfique pour la détection des états d'hypercoagulables chez le chien. / Current coagulation testing in veterinary medicine fails to reliably and consistently identify hypercoagulable states. Traditional coagulation assays (e.g. PT, PTT, antithrombin, D-dimers, etc.) only assess a small portion of the process of coagulation. Global coagulation assays, including thromboelastography and thrombin generation, allow a more complete evaluation of hemostasis, including the balance of procoagulant versus anticoagulant blood components. However, these tests still have shortcomings in terms of their diagnostic utility. The study presented in this thesis investigates modifications to conventional thrombin generation techniques as an attempt to better identify hypercoagulable states in sick dogs. More specifically, we aimed to compare the in vitro effects of thrombomodulin and dalteparin on thrombin generation performed in healthy dogs and dogs presented with illnesses associated with hypercoagulable states. We hypothesized that addition of these agents would demonstrate endogenous anticoagulant deficiencies (i.e. protein C and antithrombin) in sick dogs. Ten healthy dogs and ten sick dogs were selected based on history, physical examination and complete blood work. Platelet-poor plasma was obtained from citrated whole blood, aliquoted and frozen at -80 ̊Celsius for less than 18 months. Thrombin generation was measured on thawed plasma using a Calibrated Automated Thrombogram assay. Measurements were performed without anticoagulant and with the addition of thrombomodulin and dalteparin at three dilutions (0.2, 0.4 and 0.6 U/ml). Endogenous thrombin potential (ETP), lag time (Lag time), time to peak (TTPeak) and peak thrombin generation (Peak) were recorded and analyzed using a mixed linear model (p < 0.05). No difference was observed between the groups before the addition of anticoagulants. The addition of thrombomodulin and dalteparin significantly decreased ETP and Peak thrombin generation in both groups. However, the effects of both anticoagulants were significantly less in the diseased group compared to the healthy group, except for Peak at the highest dalteparin concentrations (0.4 U/ ml and 0.6 U/ml). The addition of thrombomodulin and dalteparin to plasma in vitro highlights an endogenous anticoagulant deficiency in this population of sick dogs, by failing to reduce thrombin generation to the same extent as in healthy dogs. Therefore, performing thrombin generation with and without the addition of exogenous anticoagulants might be beneficial for the detection of hypercoagulable states in the dog.