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Integrated time and cost-saving analysis of proteolytic activity and inhibitory provision of patient's plasma as the basis for prognosis of acute, subacute and chronic thrombo-haemorrhage complications in inflammation

Objectif


As a result of this proposal research the controversial subject regarding the stability (firmness) of the El-a 1PI complex in human plasma is to be solved.
The answer of value will be received, if the El-a1PI complex, due to its instability, is the reason of the key plasma proteins inactivation, or this inactivation is a result of imbalance between the low level of a1PI and high leucocyte elastase activity.
The proposal investigation lies in the range of the Fourth Framework Programme - under the heading of "Measurement and testing", 8.3 Health and safety, as far as the proposed measurement method is to improve the comparability of results used for in vitro diagnostic and therapeutic purposes, and the results of this method have the valid reference data used in decision-making in the clinical field.
The biochemical diagnosis of severe inflammation with disseminated processes, such as extensive trauma, peritonitis and septicaemia, predicting possible complications (acute respiratory distress syndrome, disseminated intravascular coagulation and multiple organ failure), is not properly possible in clinical laboratories of NIS nowadays. Meanwhile, there is a good reason to assume that the most destructive neutrophil enzyme, which disturbs the regulation of processes of blood coagulation and fibrinolysis, is leucocyte elastase (El). Some laboratories of CCE use the immunoenzyme (ELISA) method for estimation the concentration of the complex of leucocyte elastase with its main plasma inhibitor - alPI. This method has been suggested and developed in the beginning of 80th by researchers of Munich University, and is designed for the control of neutrophil activation and their degranulation. But this method has not become commonly accepted in clinical laboratories of both CCE and NIS because of high cost and the long-term execution (not less than 4 hours, while DIC-syndrome may develop in 1-2 hours). The high cost is of particular disadvantage for laboratories of NIS because of their financial difficulties.
Taking into account the above objects, a cheap and time-saving spectrophotometer method for evaluation of leucocyte elastase activity from complex El-alPI was elaborated at the Department of Biochemistry of Russian Medical Academy for Postgraduate Education in 1992. However, in the first place, this method needs the adaptation for clinical laboratory use, and, second, its results have to be compared with the results of ELISA of El-a1PI complex.
The very important aim of this proposal is to determine the concentration (activity) of the main coagulating and fibrinolytic protein factors, such as proteins of contact phase, in the patients' plasma with severe inflammation, and to demonstrate the correlation of their concentration (activity) with elastase activity, estimated by the new method. It is possible that the outstripping of the peak of elastase activity in severe inflammatory diseases will stimulate the search for special elastase inhibitors to prevent the thrombo-haemorrhage complications.

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Coordinateur

Ludwig-Maximilians-Universität München
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Adresse
Nussbaumstraße 20
80336 München
Allemagne

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