Impairment of fibrinolysis by streptokinase, urokinase and recombinant tissue-type plasminogen activator in the presence of radiographic contrast agents

OBJECTIVES: The purpose of this study was to determine whether an adverse interaction exists between radiographic contrast agents and thrombolytic drugs. BACKGROUND: Coronary thrombosis may occur in the setting of unstable angina and after coronary angioplasty. However, the use of thrombolytic drugs in the setting of unstable angina has not been beneficial and, in one large trial of angioplasty in patients with unstable angina, was associated with an increased incidence of ischemic complications and abrupt closure. The reasons for these results are not clear. Coronary arteriography was performed in many of these trials, and it is known that fibrin structure and assembly are altered by radiographic contrast agents. METHODS: Blood samples were obtained from patients before (n = 25) and after (n = 20) angiography using iohexol. Blood samples obtained before angiography were tested for response to streptokinase (10 and 100 IU/ml), urokinase (100, 200 and 500 IU/ml) and recombinant tissue-type plasminogen activator (rt-PA) (100 and 1,000 IU/ml) and the results measured. Iohexol, diatrizoate or ioxaglate (4% by volume) was added to separate aliquots of the baseline sample, and the test was repeated. Blood samples obtained after angiography were tested in a similar manner. RESULTS: The onset of lysis at baseline by rt-PA at 1,000 IU/ml occurred at 72 +/- 8.2 s (mean +/- SD) and was markedly delayed in the presence of diatrizoate (527 +/- 181.7 s, p < 0.001) or iohexol (460 +/- 197.0 s, p < 0.001) but not ioxaglate. At 100 IU/ml, there was no lysis detected with rt-PA after the addition of any contrast agent. The addition of a contrast agent caused similar delays in the onset of lysis by urokinase and streptokinase; similar to rt-PA, the effect was smaller at higher concentrations of drug. In vivo blood samples obtained from the patient after angiography showed delays in the onset of lysis by rt-PA and urokinase but not streptokinase. CONCLUSIONS: These data demonstrate that radiographic contrast agents impede fibrinolysis. This previously undescribed interaction was demonstrated using an in vitro test system, but these findings may have clinical relevance when thrombolytic drugs are used at the time of angiography

Micro-elastometry on whole blood clots using actuated surface-attached posts (ASAPs)

We used magnetically actuatable micro-post arrays to measure blood clot elasticity for blood clotting diagnostics

Exploring the gap between dynamic and constraint-based models of metabolism

Systems biology provides new approaches for metabolic engineering through the development of models and methods for simulation and optimization of microbial metabolism. Here we explore the relationship between two modeling frameworks in common use namely, dynamic models with kinetic rate laws and constraint-based flux models. We compare and analyze dynamic and constraint-based formulations of the same model of the central carbon metabolism of E. coli. Our results show that, if unconstrained, the space of steady states described by both formulations is the same. However, the imposition of parameter-range constraints can be mapped into kinetically feasible regions of the solution space for the dynamic formulation that is not readily transferable to the constraint-based formulation. Therefore, with partial kinetic parameter knowledge, dynamic models can be used to generate constraints that reduce the solution space below that identied by constraint-based models, eliminating infeasible solutions and increasing the accuracy of simulation and optimization methods.This research was supported by PhD Grants SFRH/BD/35215/2007 and SFRH/BD/25506/2005 from the Fundacao para a Ciencia e a Tecnologia (FCT) and the MIT-Portugal Program through the project "Bridging Systems and Synthetic Biology for the Development of Improved Microbial Cell Factories" (MIT-Pt/BS-BB/0082/2008)

Kinetic Studies of a Doubly Bound Red Cell Antigen-Antibody System

The Polybrene method for detection of red cell antibodies which utilizes continuous flow equipment was modified so that kinetic studies could be performed on red cell antibodies doubly bound between adjacent red cells. In the anti-Rh(o)-Rh(o) erythrocyte system, deaggregation by temperature was studied over an antibody concentration range of from approximately 1 to 500 antibody molecules per erythrocyte, a residence time range of approximately eightfold, and a temperature range of from 10 to 55°C. The rate of dissociation of antigen-antibody complex, as determined from deaggregation of antibody-dependent red cell aggregates, was found to be of apparent zero order. The apparent activation energy for the antigen-antibody reaction under the experimental conditions was determined and found to be higher than would be expected for singly bound antigen-antibody systems. Possible explanations are considered for these findings in terms of an antigen-antibody bond-breaking model

Micro-elastometry on whole blood clots using actuated surface-attached posts (ASAPs)

We present a novel technology for microfluidic elastometry and demonstrate its ability to measure stiffness of blood clots as they form. A disposable micro-capillary strip draws small volumes (20 μL) of whole blood into a chamber containing a surface-mounted micropost array. The posts are magnetically actuated, thereby applying a shear stress to the blood clot. The posts’ response to magnetic field changes as the blood clot forms; this response is measured by optical transmission. We show that a quasi-static model correctly predicts the torque applied to the microposts. We experimentally validate the ability of the system to measure clot stiffness by correlating our system with a commercial thromboelastograph. We conclude that actuated surface-attached post (ASAP) technology addresses a clinical need for point-of-care and small-volume elastic haemostatic assays