Abstract:
PURPOSE:The study sought to synthesize anionic peptide-conjugated tissue plasminogen activator (tPA) for its targeted/triggered delivery, where tPA's activity would be masked in the circulation and regenerated at the thrombus site by a commonly used anticoagulant, heparin, to minimize tPA associated bleeding complications. METHODS:tPA was conjugated to Polyglutamate, and the activity of oligoanion-modified tPA was tested by fibrinolytic assay. Separately human serum albumin (HSA) was conjugated to protamine and the formation of its electrostatic complex with anionic peptide was monitored by Förster Resonance Energy Transfer (FRET). The masking of tPA-activity via steric hindrance created by albumin, and subsequent regeneration with therapeutic dose of heparin was tested by enzymatic assay. Stability of 'camouflaged-tPA' was determined in human plasma. Using fluorescence microscope, binding of camouflaged-tPA with activated platelets was monitored. Heparin modulated clot-lysis was evaluated in human blood clot. RESULTS:The anionic tPA retained ~97% activity of the unmodified-tPA. FRET experiments confirmed the electrostatic interaction between polyglutamate and protamine which was subsequently reversed by heparin. Complexation with HSA-protamine masked ~60% of tPA activity which was fully regenerated by heparin. The complex retained its prodrug character in human plasma after incubation at 37°C. Fluorescence microscopic study confirmed binding of the construct with activated platelets. In lysing human clot, the camouflage could mask tPA-activity until it was triggered at a heparin level of 0.4U/mL. CONCLUSION:Oligoanion-modified tPA could be used for targeted/triggered delivery where its enzymatic activity could be masked by HSA-protamine conjugate and successfully regenerated by therapeutic dose of heparin.
journal_name
Thromb Resjournal_title
Thrombosis researchauthors
Absar S,Choi S,Ahsan F,Cobos E,Yang VC,Kwon YMdoi
10.1016/j.thromres.2012.11.030subject
Has Abstractpub_date
2013-03-01 00:00:00pages
e91-9issue
3eissn
0049-3848issn
1879-2472pii
S0049-3848(12)00851-1journal_volume
131pub_type
杂志文章abstract::A recently developed platelet aggregometer using a laser light scattering method is capable of monitoring the increase in size of small-sized platelet aggregates (diameter 9-25 microm), which cannot be detected with the conventional methods. Whether coronary spasm can cause platelet aggregation in the coronary circula...
journal_title:Thrombosis research
pub_type: 临床试验,杂志文章
doi:10.1016/s0049-3848(01)00333-4
更新日期:2001-09-01 00:00:00
abstract::Fasting plasma homocysteine level and the related clinical findings were analysed in 240 consecutive patients with venous thromboembolism. Hyperhomocysteinemia, defined as a plasma level above 20 micromol/l (corresponding to the percentile 95th in the controls), was present in 11.2% of the patients. Plasma homocystein...
journal_title:Thrombosis research
pub_type: 杂志文章
doi:10.1016/s0049-3848(02)00096-8
更新日期:2002-04-15 00:00:00
abstract::Annexin V is a human phospholipid binding protein (M(r) 36,000) that binds with high affinity to activated platelets in vitro. We studied the biodistribution and thrombus binding of annexin V in rabbit and swine models of fully occlusive arterial thrombi formed 1-2 h prior to injection of annexin V. Iodinated annexin ...
journal_title:Thrombosis research
pub_type: 杂志文章
doi:10.1016/0049-3848(94)90224-0
更新日期:1994-09-01 00:00:00
abstract::Recent studies have demonstrated that neutrophilic granulocytes and platelets have the capability to interact with each other. Either type of cell may release soluble factors, which either activate the other or increase its response to separate stimulating agents. Direct contact between the two types of cell generally...
journal_title:Thrombosis research
pub_type: 杂志文章,评审
doi:10.1016/s0049-3848(10)80002-7
更新日期:1994-01-01 00:00:00
abstract:INTRODUCTION:Severe thrombocytopenia (≤50×10(9) platelets/L) is often the consequence of hematological malignancies and intensive chemotherapy. The risk of clinically significant bleeding is increased in these patients, despite the use of prophylactic platelet transfusions. The fact that there is no clear correlation b...
journal_title:Thrombosis research
pub_type: 杂志文章
doi:10.1016/S0049-3848(16)30125-6
更新日期:2016-04-01 00:00:00
abstract::This prospective randomized study investigated the influence of normotensive and hypotensive general anesthesia on platelet aggregability, intraoperative blood loss and parameters of plasmatic coagulation during extensive orthognathic surgery. A total of 30 patients were randomly allocated for either normotensive anes...
journal_title:Thrombosis research
pub_type: 临床试验,杂志文章,随机对照试验
doi:10.1016/s0049-3848(01)00316-4
更新日期:2001-08-01 00:00:00
abstract::The purpose of this study was to determine if the results obtained in platelet function tests and whole blood perfusions are associated with those in platelet function analyser (PFA)-100. We used collagen type I monomers and fibrils to analyse the distinct roles of glycoprotein (GP) Ia/IIa and other collagen receptors...
journal_title:Thrombosis research
pub_type: 杂志文章
doi:10.1016/s0049-3848(01)00283-3
更新日期:2001-07-15 00:00:00
abstract::The abnormal Antithrombin-III (AT-III) Toyama has no ability to interact with heparin in the mast cells or heparin-like glycosaminoglycans on the surface of vascular endothelial cells, so that it has low heparin cofactor activity. In the present study, we have measured the binding capacity of 125I-labeled AT-III to cu...
journal_title:Thrombosis research
pub_type: 杂志文章
doi:10.1016/0049-3848(88)90170-3
更新日期:1988-04-01 00:00:00
abstract::Homocystinuria due to cystathionine-beta-synthase deficiency (CBS-def-HOCY) initially often presents with vascular disorders, e.g. thromboembolic events. The measurement of vascular endothelial markers in plasma could help to assess endothelial damage. We determined von Willebrand factor (measured as Ristocetincofacto...
journal_title:Thrombosis research
pub_type: 杂志文章
doi:10.1016/0049-3848(95)90867-f
更新日期:1995-01-01 00:00:00
abstract::The effects of ticlopidine treatment (250 mg b.i.d. for 21 months) on fibrinogen and other rheological variables, as compared to placebo, were studied in 44 patients with intermittent claudication due to peripheral arterial occlusive disease. Blood samples were collected every 3 months during this double-blind, random...
journal_title:Thrombosis research
pub_type: 临床试验,杂志文章,随机对照试验
doi:10.1016/0049-3848(88)90134-x
更新日期:1988-12-15 00:00:00
abstract:BACKGROUND:The usefulness of coagulation tests performed before epidural analgesia for surgery or to alleviate labour pain is controversial. The aims of this study were: (1) to evaluate the prevalence of abnormal tests in a large cohort of healthy pregnant women and their association with epidural hematoma; (2) to asse...
journal_title:Thrombosis research
pub_type: 杂志文章
doi:10.1016/j.thromres.2011.02.023
更新日期:2011-07-01 00:00:00
abstract::Platelet aggregate size was determined with a newly-developed platelet aggregometer, PA-100 (KOWA), which can quantitatively evaluate the size and number of platelet aggregates by means of the particle counting method based upon light scattering. Epinephrine-induced platelet aggregation consists of two phases, the for...
journal_title:Thrombosis research
pub_type: 杂志文章
doi:10.1016/0049-3848(96)00026-6
更新日期:1996-03-01 00:00:00
abstract::Patients with active cancer have a heightened risk of venous thromboembolism (VTE). This risk is further increased by the initiation of chemotherapy. Although previous studies have suggested that the use of parenteral thromboprophylaxis in all ambulatory cancer patients receiving chemotherapy significantly decreases t...
journal_title:Thrombosis research
pub_type: 杂志文章
doi:10.1016/j.thromres.2018.01.018
更新日期:2018-04-01 00:00:00
abstract::Bovine hide collagen dispersions were swollen in the pH range 1.6-7.0, treated with glutaraldehyde, and dialyzed to neutral pH. The intensity with which these collagens reacted with human platelets in plasma was studied by aggregometry and scanning electron microscopy. Collagen swollen at a pH below 4.25 +/- 0.30 and ...
journal_title:Thrombosis research
pub_type: 杂志文章
doi:10.1016/0049-3848(89)90463-5
更新日期:1989-07-01 00:00:00
abstract:BACKGROUND:Although patients undergoing coronary stenting routinely receive dual antiplatelet treatment to reduce the risk of stent thrombosis, this undesired event still occurs. A suboptimal response to clopidogrel treatment (low responders) has been suggested to contribute to stent thrombosis. In the present study, p...
journal_title:Thrombosis research
pub_type: 杂志文章
doi:10.1016/j.thromres.2004.07.007
更新日期:2005-01-01 00:00:00
abstract::Mice were implanted subcutaneously with a pellet containing 0.5 mg estradiol or with a placebo. Eight to 12 days later platelet aggregation was produced in mesenteric arterioles by injuring their endothelium in vivo with a noxious light/dye stimulus. The time between the onset of the noxious stimulus and the appearanc...
journal_title:Thrombosis research
pub_type: 杂志文章
doi:10.1016/0049-3848(85)90220-8
更新日期:1985-08-01 00:00:00
abstract::Thrombus formation was electrically induced in the left common carotid artery of rats with preoccluded both vertebral and right carotid arteries. Following thrombus induction, animals became unresponsive and lost their righting reflex. The loss of righting reflex was observed when cortical tissular pO2 reached 55 +/- ...
journal_title:Thrombosis research
pub_type: 杂志文章
doi:10.1016/0049-3848(87)90340-9
更新日期:1987-10-01 00:00:00
abstract:BACKGROUND:There is a substantial local release of tissue-type plasminogen activator (t-PA) in the splanchnic vascular bed, and this release is increased at high sympathetic tone. Coronary t-PA release is also significant, and this release increases during cardiac nerve stimulation and during reperfusion after 10 min o...
journal_title:Thrombosis research
pub_type: 杂志文章
doi:10.1016/j.thromres.2009.10.004
更新日期:2010-03-01 00:00:00
abstract:BACKGROUND AND AIM:Recent studies have documented that elevation of C-reactive protein (CRP) levels after percutaneous coronary intervention (PCI) have been predictive of adverse outcome. This study was performed to test the hypothesis that preprocedural use of naproxen sodium is associated with a reduction in the exte...
journal_title:Thrombosis research
pub_type: 杂志文章,随机对照试验
doi:10.1016/j.thromres.2006.11.003
更新日期:2007-01-01 00:00:00
abstract::The presence of Factor XIII subunit a was demonstrated in human monocytes by immunoperoxidase staining using specific antisera against Factor XIII and its subunits. This finding was verified by immunobiochemical techniques, as well. In an immunoblotting system after SDS polyacrylamide gel electrophoresis of denatured ...
journal_title:Thrombosis research
pub_type: 杂志文章
doi:10.1016/0049-3848(85)90069-6
更新日期:1985-02-01 00:00:00
abstract:OBJECTIVES:Dabigatran etexilate is widely used for stroke prevention in the patients with atrial fibrillation. The anticoagulation activity of dabigatran is not necessary monitored in routine clinical practice. We aimed to study the effect of dabigatran on thrombin generation (TG) and coagulation assays in rabbit and h...
journal_title:Thrombosis research
pub_type: 杂志文章
doi:10.1016/j.thromres.2018.03.012
更新日期:2018-05-01 00:00:00
abstract::Three collagenases were purified from the culture medium of human skin fibroblasts using ammonium sulfate fractionation, ion-exchange chromatography and gel filtration. The cationic collagenase had a molecular weight of 42,000; two anionic collagenases had molecular weights of 63,000 and 115,000. Preincubation of the ...
journal_title:Thrombosis research
pub_type: 杂志文章
doi:10.1016/0049-3848(84)90211-1
更新日期:1984-07-15 00:00:00
abstract:INTRODUCTION:Unfractionated heparin (UFH) potentiates platelet aggregation induced by some agonists. P2Y12 and P2Y1 receptors play a major role in amplifying platelet aggregation. We assessed the ability of cangrelor, a selective P2Y12 antagonist, A2P5P, a selective P2Y1 antagonist, and aspirin to block the potentiatin...
journal_title:Thrombosis research
pub_type: 杂志文章
doi:10.1016/j.thromres.2004.09.004
更新日期:2005-01-01 00:00:00
abstract::Ethamsylate possesses antihemorrhagic properties, but whether or not it directly activates blood platelets is unclear. Here we investigated the platelet activation potential of ethamsylate, by measuring membrane P-selectin expression with flow cytometry in human whole blood and also by immunofluorescence imaging of is...
journal_title:Thrombosis research
pub_type: 杂志文章
doi:10.1016/s0049-3848(02)00353-5
更新日期:2002-09-15 00:00:00
abstract::The effects of sodium salicylate (SS) on ADP-induced platelet aggregation and on a metabolism of calcium in platelets were studied, using gel-filtrated platelets (GFP). SS inhibited dose-responsively ADP-induced aggregation in the presence of fibrinogen and Ca2+. It was found that extracellular 45Ca2+ was rapidly take...
journal_title:Thrombosis research
pub_type: 杂志文章
doi:10.1016/0049-3848(85)90232-4
更新日期:1985-08-01 00:00:00
abstract::Blood platelets have recently been shown to express PYK2, a nonreceptor tyrosine kinase belonging to the FAK gene family. In this study, we examined the involvement of protein kinase C (PKC) in PYK2-related responses in human platelets. While PYK2 tyrosine phosphorylation induced by thrombin was inhibited by preincuba...
journal_title:Thrombosis research
pub_type: 杂志文章
doi:10.1016/s0049-3848(98)00188-1
更新日期:1999-03-15 00:00:00
abstract::The presence of one acidifying Cl-/HCO3- exchange mechanism in human platelets has not been previously reported. This paper demonstrates that this mechanism does function and that it increases its activity after stimulation with thrombin. On resuspension of BCECF-loaded platelets in a chloride-free medium (gluconate r...
journal_title:Thrombosis research
pub_type: 杂志文章
doi:10.1016/j.thromres.2004.11.012
更新日期:2005-01-01 00:00:00
abstract::An anticoagulant activity was identified and isolated from the leaves of a West African plant, Aspilia africana by gel filtration on Sephadex G-100. The anticoagulant factor had an apparent molecular weight of approximately 60,000 d. Upon incubation with plasma, it prolonged the partial thromboplastin time, prothrombi...
journal_title:Thrombosis research
pub_type: 杂志文章
doi:10.1016/0049-3848(87)90455-5
更新日期:1987-08-15 00:00:00
abstract:BACKGROUND:Clinical and epidemiological studies have associated selective COX-2 inhibitors with an increased risk of cardiovascular events. There are no clinical studies on the possible effects of these drugs on secondary hemostasis. The hypothesis for this study is that the use of selective COX-2 inhibitors could affe...
journal_title:Thrombosis research
pub_type: 临床试验,杂志文章,随机对照试验
doi:10.1016/j.thromres.2009.01.017
更新日期:2009-06-01 00:00:00
abstract:INTRODUCTION:Antiphospholipid syndrome (APS) is a pro-thrombotic autoimmune disease that affects different vascular beds, with potential risk for recurrence. Systemic lupus erythematosus (SLE), specific autoantibodies profile and atherogenic disorders have been described as risk factors for the occurrence of first thro...
journal_title:Thrombosis research
pub_type: 杂志文章
doi:10.1016/j.thromres.2015.10.029
更新日期:2015-12-01 00:00:00