Abstract：
:The three main components involved in thrombosis and haemostasis are thrombin, platelets, and plasmin. Almost all inhibitors of thrombosis are focused either on the inhibition of thrombin or on the inhibition of platelets. We designed a construct using the fibrinolytic activity of staphylokinase, fused via a cleavable linker to an antithrombotic peptide of 29 amino acids. The peptide was designed to include three inhibitory regions: (1) the Arg-Gly-Asp (RGD) amino acid sequence to prevent fibrinogen binding to platelets; (2) a part of fibrinopeptide A, an inhibitor of thrombin; and (3) the tail of hirudin, a potent direct antithrombin. The amino acid sequence of the 29 amino acid peptide was reverse translated, and the gene was chemically synthesised and cloned into an expression vector as a 3' fusion to the staphylokinase gene. Gene expression was induced in E. coli Top 10 cells and the fusion protein, designated PLATSAK, was purified using metal affinity chromatography. The purified fusion protein significantly lengthened the activated partial thromboplastin time and thrombin time and inhibited the amidolytic activity of thrombin. The fibrinolytic activity was almost equal to that of recombinant staphylokinase as measured with a thrombelastograph. Platelet aggregation was not markedly inhibited by PLATSAK, probably due to the unfavourable three dimensional structure, with the Arg-Gly-Asp sequence buried inside. Our results confirm that it is feasible to design and produce a hybrid multifunctional protein that targets various components of the haemostatic process.
journal_name
Thromb Resjournal_title
Thrombosis researchauthors
van Zyl WB,Pretorius GH,Hartmann M,Kotzé HFdoi
10.1016/s0049-3848(97)00277-6subject
Has Abstractpub_date
1997-12-01 00:00:00pages
419-26issue
5eissn
0049-3848issn
1879-2472pii
S0049-3848(97)00277-6journal_volume
88pub_type
杂志文章abstract：:Fibrin is found in most solid tumours, and there is speculation regarding its role in tumour invasion and metastasis. An assay to quantitate fibrin levels in tissues would be a useful preliminary tool in assessing the above. Such an assay to quantitatively detect levels of fibrin in various types of canine tumour was ...
journal_title：Thrombosis research
pub_type： 杂志文章
doi：10.1016/s0049-3848(99)00054-7
更新日期：1999-10-01 00:00:00
abstract：:Chronic thromboembolic pulmonary hypertension (CTEPH) is the most severe long term complication of acute pulmonary embolism (PE). Untreated, CTEPH is associated with a very poor prognosis and high risk of mortality, although curation can be achieved by surgical removal of the obstructive endothelialised thromboemboli ...
journal_title：Thrombosis research
pub_type： 杂志文章,评审
doi：10.1016/j.thromres.2016.12.026
更新日期：2017-03-01 00:00:00
abstract：:Sandwich ELISA systems were developed to measure human von Willebrand factor (vWF). In one system the microtitre plates were coated with goat F(ab')2 to vWF. The F(ab')2 different molecular forms in the soluble phase. VWF antigen bound by the F(ab')2 antibody was subsequently determined by using horse-radish peroxidas...
journal_title：Thrombosis research
pub_type： 杂志文章
doi：10.1016/0049-3848(86)90047-2
更新日期：1986-07-01 00:00:00
abstract：BACKGROUND:Cancer patients have an increased risk of thrombosis. Tissue factor (TF) antigen and TF activity associated with microparticles in plasma are elevated in patients with various types of cancer. Of these two measurements, TF activity is considered superior to TF antigen levels because the activity more closely...
journal_title：Thrombosis research
pub_type： 杂志文章
doi：10.1016/j.thromres.2007.12.023
更新日期：2008-01-01 00:00:00
abstract：BACKGROUND:The quality of treatment with warfarin is mainly assessed by the time in therapeutic range (TTR) in patients with mechanical heart valve prosthesis (MHV). Our aim was to evaluate if International Normalized Ratio (INR) variability predicted a combined endpoint of thromboembolism, major bleeding and death bet...
journal_title：Thrombosis research
pub_type： 杂志文章
doi：10.1016/j.thromres.2015.10.044
更新日期：2015-12-01 00:00:00
abstract：BACKGROUND:Although risk of hospital-associated venous thromboembolism (HA-VTE) differs between critically and non-critically ill children, studies to date have not led to distinct, pragmatic risk scores. OBJECTIVE:To determine risk factors for HA-VTE in critically ill children not undergoing cardiothoracic surgery, i...
journal_title：Thrombosis research
pub_type： 杂志文章
doi：10.1016/j.thromres.2015.04.036
更新日期：2015-10-01 00:00:00
abstract：BACKGROUND:Acquired von Willebrand Syndrome is a rare bleeding disorder, which arises in individuals with no personal or family history of bleeding, associated with lymphoproliferative and myeloproliferative disorders or other diseases. AIM:To develop a two-step approach assay to detect autoantibodies against VWF and ...
journal_title：Thrombosis research
pub_type： 杂志文章
doi：10.1016/j.thromres.2014.09.020
更新日期：2014-12-01 00:00:00
abstract：:Diagnosis and risk stratification of patients with acute coronary syndromes remain extremely important in order to avoid unnecessary hospitalizations on the one hand, and to improve prognosis of these patients on the other. For diagnosis of acute coronary syndromes, an ECG should be obtained at rest, troponin T and I ...
journal_title：Thrombosis research
pub_type： 杂志文章,评审
doi：10.1016/s0049-3848(01)00298-5
更新日期：2001-09-30 00:00:00
abstract：OBJECTIVE:To compare rates of VTE occurrence in obese versus non-obese hospitalized patients who received UFH 5000 units subcutaneously q8 h. MATERIALS AND METHODS:This was a retrospective cohort study conducted in an academic medical center in the United States. Consecutive adult patients who were hospitalized during...
journal_title：Thrombosis research
pub_type： 杂志文章
doi：10.1016/j.thromres.2018.07.027
更新日期：2018-09-01 00:00:00
abstract：:Using high-resolution real-time two-dimensional ultrasound, we have investigated the role of red cells in the growth of already established platelet aggregates under controlled flow conditions. Platelet rich plasma (PRP) was circulated in vitro in horizontally and vertically arranged tubing at mean shear rate ranging ...
journal_title：Thrombosis research
pub_type： 杂志文章
doi：10.1016/0049-3848(84)90376-1
更新日期：1984-10-01 00:00:00
abstract：:Heparin cofactor II (HCII) is a specific thrombin inhibitor; its inhibitory activity is stimulated by heparin (Hep) and dermatan sulfate (DS). Vitronectin (VN), a heparin binding adhesive glycoprotein present in plasma and extracellular matrix, has been shown to decrease the stimulatory effect of Hep but not of DS on ...
journal_title：Thrombosis research
pub_type： 杂志文章
doi：10.1016/0049-3848(92)90310-7
更新日期：1992-06-01 00:00:00
abstract：:We investigated the relationship between the procoagulant protease-inhibitory activity and the N-glycan structures in urinary protein C inhibitor (uPCI) by sequential exoglycosidase digestions based on the N-glycan structures elucidated in this report. uPCI was glycosylated on the three potential N-glycosylation sites...
journal_title：Thrombosis research
pub_type： 杂志文章
doi：10.1016/s0049-3848(01)00342-5
更新日期：2001-10-01 00:00:00
abstract：:Fibrillar collagen endows the normal aortic wall with significant stiffness and strength and similarly plays important roles in many disease processes. For example, because of the marked loss of elastic fibers and functional smooth cells in aortic aneurysms, collagen plays a particularly important role in controlling ...
journal_title：Thrombosis research
pub_type： 杂志文章
doi：10.1016/j.thromres.2012.04.009
更新日期：2012-09-01 00:00:00
abstract：INTRODUCTION:This study determines the impact of tissue factor (TF)-signaling on the extrinsic pathway of apoptosis in cancer cells and propose death associated protein kinase-1 (DAPK1) as a novel key regulator. MATERIALS AND METHODS:In MDA-MB-231 breast and PC3 prostate cancer cells, mRNA levels were analyzed by real...
journal_title：Thrombosis research
pub_type： 杂志文章
doi：10.1016/j.thromres.2010.11.015
更新日期：2011-02-01 00:00:00
abstract：INTRODUCTION:Although recurrent venous thromboembolism is a known risk factor for chronic thromboembolic pulmonary hypertension (CTEPH), the prevalence of CTEPH after recurrent pulmonary embolism (PE) is not clear. MATERIALS AND METHODS:A cohort screening study was performed to clarify the prevalence of echocardiograp...
journal_title：Thrombosis research
pub_type： 杂志文章
doi：10.1016/j.thromres.2011.07.045
更新日期：2011-12-01 00:00:00
abstract：OBJECTIVES:In this randomized management study, we examined the safety of withholding anticoagulation on the basis of negative impedance plethysmography (IPG) compared to negative contrast venography (CV) in symptomatic patients with a first episode of clinically suspected deep vein thrombosis (DVT), and we determined ...
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)00222-5
更新日期：2001-04-01 00:00:00
abstract：:It has been suggested that unstable angina at rest, like acute myocardial infarction, might be associated with a thrombotic process. In order to study the hypothesis that myocardial ischemia during exercise could also be associated with an activation of blood coagulation and/or fibrinolysis, we investigated the presen...
journal_title：Thrombosis research
pub_type： 杂志文章
doi：10.1016/0049-3848(92)90222-v
更新日期：1992-01-01 00:00:00
abstract：BACKGROUND:Tissue factor pathway inhibitor-2 (TFPI-2) regulates matrix metalloproteinases activation and extracellular matrix degradation. Over-expression of TFPI-2 enhances atherosclerotic plaque stability. The aim of this study is to investigate the effect of conditional knockout (KO) of TFPI-2 in vascular endothelia...
journal_title：Thrombosis research
pub_type： 杂志文章
doi：10.1016/j.thromres.2015.11.010
更新日期：2016-01-01 00:00:00
abstract：INTRODUCTION:Coagulation factor XIII (FXIII) is a fibrin-stabilizing factor, which contributes to hemostasis, wound healing, and maintenance of pregnancy. Accordingly, patients with congenital FXIII deficiency manifest a life-long bleeding tendency, abnormal wound healing and recurrent miscarriage. In order to understa...
journal_title：Thrombosis research
pub_type： 杂志文章
doi：10.1016/j.thromres.2012.05.003
更新日期：2012-09-01 00:00:00
abstract：:Selective serotonin reuptake inhibitors (SSRIs) have been reported to reduce platelet aggregation induced by ADP. ADP induces platelet aggregation through two purinergic receptors P2Y1 and P2Y12. We characterized the inhibitory properties of SSRIs on ADP-induced platelet aggregation and investigated the effects of SSR...
journal_title：Thrombosis research
pub_type： 杂志文章
doi：10.1016/j.thromres.2013.02.007
更新日期：2013-04-01 00:00:00
abstract：:Separation of plasma, erythrocytes, and platelets was studied in T-shaped bifurcations prepared by a casting technique. The diameters of the side and main channel were 100 microns and 200 microns, respectively. Small blood samples (about 20 microliters) were collected from the side channel for determination of erythro...
journal_title：Thrombosis research
pub_type： 杂志文章
doi：10.1016/0049-3848(87)90314-8
更新日期：1987-03-01 00:00:00
abstract：:An in vivo rabbit model of venous thrombosis which includes physiological blood flow was used to compare the efficacy of the potent and specific factor Xa inhibitor recombinant tick anticoagulant peptide (rTAP) with standard heparin in the prevention of venous thrombus formation. In anesthetized rabbits, an autologous...
journal_title：Thrombosis research
pub_type： 杂志文章
doi：10.1016/0049-3848(93)90200-8
更新日期：1993-08-15 00:00:00
abstract：:We determined the extent and duration of activation of coagulation during the first 24 hours after carotid endarterectomy. Serial blood samples were collected before, during and after surgery from 10 patients with severe stenosis (> 70%) undergoing carotid endarterectomy with heparinization. Platelet poor plasmas prep...
journal_title：Thrombosis research
pub_type： 杂志文章
doi：10.1016/0049-3848(94)90257-7
更新日期：1994-08-15 00:00:00
abstract：:Four novel polymorphisms were identified in the fibrinogen gene cluster. Three of them were localized in the promoter regions of the Aalpha-chain (alpha -128 C/G, alpha -58 G/A) or the gamma-chain (gamma -239 A/G) gene, while the remaining one was identified in intron 9 of the gamma-chain gene (gamma 7792 C/T). Genoty...
journal_title：Thrombosis research
pub_type： 杂志文章
doi：10.1016/s0049-3848(01)00329-2
更新日期：2001-08-15 00:00:00
abstract：:Von Willebrand Factor is a multimer produced by endothelial cells and megakaryocytes, being stored in intracellular organelles, such as the Weibel-Palade bodies and alpha-granules in endothelial cells and platelets, respectively. This molecule acts as a carrier protein for factor VIIIc, involved in the intrinsic pathw...
journal_title：Thrombosis research
pub_type： 杂志文章
doi：10.1016/s0049-3848(99)00122-x
更新日期：2000-01-15 00:00:00
abstract：:Antiphospholipid syndrome (APS) is an autoimmune disease characterized by arterial, venous or small-vessel thrombotic events, and recurrent miscarriages or fetal loss. APS diagnosis is based on the repeated detection of anti-phospholipid (PL) antibodies (Ab), typically associated with anti-β2 glycoprotein I (β2GPI)-Ab...
journal_title：Thrombosis research
pub_type： 杂志文章,评审
doi：10.1016/j.thromres.2014.11.034
更新日期：2015-02-01 00:00:00
abstract：:Acute promyelocytic leukemia (APL) has become the most curable form of acute myeloid leukemia after the advent of all-trans retinoic acid (ATRA). However, early deaths (ED) mostly due to the disease-associated coagulopathy remain the major cause of treatment failure. In particular, hemorrhagic events account for 40-65...
journal_title：Thrombosis research
pub_type： 杂志文章,评审
doi：10.1016/S0049-3848(14)50019-9
更新日期：2014-05-01 00:00:00
abstract：:We have previously reported on a microassay based on human kininogens as cysteine proteinase inhibitors (CPIs), which could quantify partially purified kininogens from different biological fluids (J Pharmacol Meth 26, 113-124, 1991). In the present study we describe a functional method that, when assuming a 1:1 stoich...
journal_title：Thrombosis research
pub_type： 杂志文章
doi：10.1016/0049-3848(96)00073-4
更新日期：1996-05-01 00:00:00
abstract：:The ability of parenteral heparin to release the capillary-bound enzymes lipoprotein lipase (LPL) and hepatic triglyceride lipase (HTGL) into the circulation is shared by several other sulphated polysaccharides. The lipase-releasing activity in rats of two unfractionated heparins has been compared with that of charact...
journal_title：Thrombosis research
pub_type： 杂志文章
doi：10.1016/0049-3848(89)90068-6
更新日期：1989-08-01 00:00:00
abstract：INTRODUCTION:Uremic bleeding frequently occurs in dialysis patients. Although its mechanism is not well characterized, acquired platelet dysfunction has been implicated in its pathogenesis. Skin bleeding time has been used to characterize platelet dysfunction in this population. However, the bleeding time is prone to e...
journal_title：Thrombosis research
pub_type： 杂志文章
doi：10.1016/j.thromres.2006.02.013
更新日期：2007-01-01 00:00:00
