Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.4.21.5 (thrombin)
 33,306 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Human blood platelets in ACD plasma were stored in sterile plastic bags for 24-96 h at the ambient temperature without agitation. No spontaneous aggregation nor bacterial contamination were noted. A progressive loss of the following parameters was seen: platelet count; ADP-, thrombin-, collagen-, and epinephrine-induced platelet aggregation; platelet factor 3 activity; reversible response to the osmotic shock; volumetric constants; amount of UV-absorbant material; 14C-5-hydroxytryptamine and 3H-adenosine uptake and release; platelet population pattern and glycogen synthesis activity. The platelet aggregation and release, the osmotic shock test, and the platelet population pattern appear to better illustrate the early changes during platelet storage and to account for the 25-42% of recirculation of 24 h stored platelets administered into thrombocytopenic patients. As stated by Murphy and Gaardner, platelets stored at 20-22 degrees C with or without agitation, although having failed to retain total functional and biochemical capacities, paradoxically seem to recuperate in vivo as shown by survival data and hemostatic effects.
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PMID:Studies on human platelets stored at 20-22 degrees C without agitation. 82 71

Degranol was chosen from the group of cytostatic drugs and Cytosar and Flucrouracil from the group of antimetabolites for investigations on the effects of cytostatics on the metabolism of thrombocytes. After isolation from blood collected on ACD fluid thromtocytes were suspended in own plasma containing EDTA and the cytostatic agent in a concentration of 6 x 10(-3)M and were subjected to incubation during 3 hours at 37 degrees C. After incubation the thrombocyte count, the levels of nucleotide compounds, and adenine nucleotides (ATP + ADP) released by thrombocytes under the action of thrombin, and the activity of dehydrogenase glucose-6-phosphate and lactic dehydrogenase were determined. The results of these determinations showed that only Degranol as a compounds belonging to the group of alkylating cytostatics caused lowering of ATP level (by about 30%) in the incubated cells. The drugs from the group of antimetabolites caused no significant changes in the studied parameters.
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PMID:[Effect of cytostatic drugs on the metabolism of incubated human blood platelets]. 125 75

Blood collected into different anticoagulants was stored in small tubes at +4 degrees C for up to 26 h. Seven blood coagulation analyses were performed under standardized conditions. High yield and stability of factor VIII:C were found for ACD and CPD-adenine. No changes could be found in the other six parameters tested. Whole blood in blood bags could be stored for 2-4 h at +4 degrees C with maximal yield of F VIII:C, with blood stored overnight the recovery was 65%. In plasma F VIII:C was stable for at least 2 h at room temperature. F VIIIR:Ag and F VIIIR:RCoF were stable in both whole blood and plasma. No activation by plasmin as measured by B beta 15-42 could be demonstrated. The initial FPA levels, reflecting thrombin activation, in the donated blood differed individually and in some blood bags very high concentrations were found. The levels of FPA were not correlated to the time for collection of a bag of blood.
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PMID:Improvement of plasma quality as raw material for factor VIII:C concentrates. Storage of whole blood and plasma and interindividual plasma levels of fibrinopeptide A. 641 86

Flow cytometry (FC) provides a reproducible investigation of cell surface antigens on platelets. The aim of this study was to elaborate appropriate protocols and to compare them with other techniques that have already been published. (1) Venipuncture with tubes containing citrate was better for the preservation of the antigenicity than using ACD tubes. The isolated platelets could not be completely distinguished from detritus and protein aggregates. Therefore a platelet concentration between 10(7) and 10(8)/ml measurement buffer was necessary to obtain a sufficient resolution by FC. (2) Isolation methods using either differential centrifugation or diluted Ficoll-Hypaque as a flotation medium provided platelets of equal purity. The method with Ficoll-Hypaque resulted in a higher number of isolated platelets than differential centrifugation. The demonstration of platelets and their antigens in whole blood without isolation gave good results provided the platelets were not activated. Activation of platelets with 1 NIH-U thrombin/l resulted in the loss of a part of the highly activated platelets because of their aggregation. (3) Comparing different concentrations of paraformaldehyde in PBS, fixation with 1% for 15 min provided the best antigen preservation for most of the antigens investigated. Isolation induced platelet activation. In order to avoid this effect, the whole anticoagulated blood was fixed with 1% paraformaldehyde for 15 min immediately after venipuncture. Then the platelets were isolated using diluted Ficoll-Hypaque. In this way, systemic activation of platelets can be detected with antibodies against glycoproteins which are translocated from the alpha-granules or lysosomes to the cell membrane. These activation markers can be determined on immediately fixed platelets (already in the whole blood) without any interference due to unspecific activation caused by the isolation procedure. (4) Platelet treatment with citric acid at pH 3, in order to remove the antigenicity of HLA-class I molecules, was sensitive to immediate fixation with paraformaldehyde in the whole blood. Fixation after isolating the platelets made it possible to demonstrate antigen stripping, and the free heavy chain, devoid of the beta 2-microglobulin, could be clearly demonstrated. (5) Using standardization beads, the average number of antigenic sites per platelet could be determined for the investigated specificities. It was shown that antibodies which have been directly conjugated or biotinylated and combined with streptavidin-phycoerythrin yielded similar results in terms of the number of antigenic binding sites while unconjugated antibodies in combination with FITC-conjugated anti-mouse-IgG led to overestimation of antigenic binding sites.
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PMID:Standardization of the flow cytometric determination of HLA class I antigens, 'platelet-specific' glycoproteins and activation markers. 776 17

A similarity-driven approach to flexible ligand docking is presented. Given a reference ligand or a pharmacophore positioned in the protein active site, the method allows inclusion of a similarity term during docking. Two different algorithms have been implemented, namely, a similarity-penalized docking (SP-DOCK) and a similarity-guided docking (SG-DOCK). The basic idea is to maximally exploit the structural information about the ligand binding mode present in cases where ligand-bound protein structures are available, information that is usually ignored in standard docking procedures. SP-DOCK and SG-DOCK have been derived as modified versions of the program DOCK 4.0, where the similarity program MIMIC acts as a module for the calculation of similarity indices that correct docking energy scores at certain steps of the calculation. SP-DOCK applies similarity corrections to the set of ligand orientations at the end of the ligand incremental construction process, penalizing the docking energy and, thus, having only an effect on the relative ordering of the final solutions. SG-DOCK applies similarity corrections throughout the entire ligand incremental construction process, thus affecting not only the relative ordering of solutions but also actively guiding the ligand docking. The performance of SP-DOCK and SG-DOCK for binding mode assessment and molecular database screening is discussed. When applied to a set of 32 thrombin ligands for which crystal structures are available, SG-DOCK improves the average RMSD by ca. 1 A when compared with DOCK. When those 32 thrombin ligands are included into a set of 1,000 diverse molecules from the ACD, DIV, and WDI databases, SP-DOCK significantly improves the retrieval of thrombin ligands within the first 10% of each of the three databases with respect to DOCK, with minimal additional computational cost. In all cases, comparison of SP-DOCK and SG-DOCK results with those obtained by DOCK and MIMIC is performed.
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PMID:Similarity-driven flexible ligand docking. 1089 86

We studied the inhibition of platelet microparticle (MP) formation and thrombin generation under high shear forces. We hypothesized that an inhibitor of the GPIb a -von Willebrand factor (vWF) interaction would be more effective in suppressing MP formation and thrombin generation than GPIIb/IIIa inhibitors. Platelet-rich plasma (PRP) anticoagulated with PPACK (D-Phe-Pro-Arg chloromethyl ketone) was exposed in a cone-and-plate viscometer (shear: 5,000 s(-1) for 5 min) in the presence of antagonists to GPIb a (the monoclonal antibody [Mab] Ib-23) or to GPIIb/IIIa (abciximab, tirofiban, eptifibatide) at their IC90 determined in platelet aggregometry with ristocetin or ADP, respectively. We used double labeling (CD41-PE and annexin-V-FITC) for flow cytometric detection of MP and their aminophospholipid exposure. Thrombin generation was measured using PRP prepared from ACD anticoagulated blood. About 40% of the thrombin generation was found to be mediated by the MP fraction of the PRP. Blockade of GPIb a with Mab Ib-23 reduced MP formation and thrombin generation by 50%, and was more effective than any GPIIb/IIIa antagonist. The combination of Mab Ib-23 with one of the GPIIb/IIIa inhibitors further reduced the MP formation to ~ 30%. The antibody also partially inhibited thrombin induced platelet aggregation. Epitope mapping suggested that Mab Ib-23 binds between the amino acids 201 and 268 of GPIb a , explaining the interference with vWF and thrombin interaction. In contrast to the commonly used GPIIb/IIIa antagonists, the blockade of GPIb a with Mab Ib-23 effectively reduces the prothrombotic MP generation and thrombin formation at shear rates typically found in arterial stenoses.
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PMID:Platelet microparticle formation and thrombin generation under high shear are effectively suppressed by a monoclonal antibody against GPIba. 1793 21

Molecular dynamics (MD) simulations followed by molecular mechanics generalized Born surface area (MM-GBSA) analyses have been carried out to study the selectivity of two neutral and weakly basic P1 group inhibitors (177 and CDA) to thrombin and trypsin. Detailed binding free energies between these inhibitors and individual protein residues are calculated by using a per-residue basis decomposition method. The analysis of the detailed interaction energies provides insight on the protein-inhibitor-binding mechanism and helps to elucidate the basis for achieving selectivity through interpretation of the structural and energetic results from the simulations. The study shows that the dominant factor of selectivity for both inhibitors is van der Waals energy, which suggests better shape complementarity and packing with thrombin. Nonpolar solvation free energy and total entropy contribution are also in favor of selectivity, but the contributions are much smaller. Binding mode and structural analysis show that 177 binds to thrombin and trypsin in a similar binding mode. In contrast, the CDA binds to thrombin and trypsin in very different modes.
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PMID:Selectivity of neutral/weakly basic P1 group inhibitors of thrombin and trypsin by a molecular dynamics study. 1868 Jan 36