• Title/Summary/Keyword: Urokinase

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Purification of Urokinase and Pro-urokinase and Comparison of their Characteristics (유로키나제와 프로유로키나제의 정제 및 특성 비교)

  • Lee, Seung-Jin;Byun, Sang-Yo
    • KSBB Journal
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    • v.14 no.6
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    • pp.724-730
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    • 1999
  • Characteristics and enzyme activity comparison was made between urokinase isolated from urine and pro-urokinase separated from CHO(Chinese Hamster Ovary) cell culture broth. Both of purified urokinase and pro-urokinase resulted 54Kd single band in electrophoresis. Urokinase which was proved as a single molecule by gel filtration showed two separated 33Kd and 21Kd bands by 2-mercaptoethanol reduction. Isoelectric forcusing resulted same pl value of 8.6 for both of them. N-terminal amino acid sequence of urokinase after 159th Ile was Ile-Gly-Gly-Glu-Phe-Thr-Thr-Ile-Glu which was different from another N-terminal sequence of Ser-Asn-Glu-Leu-His-Gln-Val-Pro-Ser-Asn. Thrombolytic activities of both of them were propotional to the enzyme concentration. Urokinase showed thrombolytic activities in an short period of reaction time. Pro-urokinase, however, showed high thrombolytic activity for 2 hours or longer period of reaction time.

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A Simplified Procedure for the Large-Scale Purification of Urokinase from Human Urine (인뇨로부터 유로키나제 대량정제공정의 단순화)

  • 정광회;선우명
    • KSBB Journal
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    • v.5 no.2
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    • pp.183-189
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    • 1990
  • An efficient method has been developed for the purification of urokinase from 1, 000 liter batches of human urine. The procedure involved precipitation of urokinase with 2mM zinc chloride, resuspension of the precipitate with 0.1M EDTA/0.5M Glycine solution, and CM-Toyopearl and benzamidine-Sepharose column chromatography. The purified urokinase was fully active and possessed a specific activity of 1.07$\times$105IU/mg. The recoveries ranged from 42 to 65% in several preparations(mean value was 51%). And the urokinase purified by this process consisted of about 13% of single chain urokinase (pro-urokinase) as evaluated by SDS-polyacrylamide gel electrophoresis in reducing condition and by S-2444 amldolytic activity under plasmin treatment.

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The Effect of Pressure Injection of Urokinase to Reverse the "No-Reflow" Phenomenon ("No-Reflow" 현상에 대한 Urokinase 압력주입의 효과)

  • Park, Dae-Hwan;May, Jr, James.W.
    • Archives of Reconstructive Microsurgery
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    • v.3 no.1
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    • pp.40-44
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    • 1994
  • Microsurgery has advanced beyond its nascent stages reaching success rates of 90% to 95%. However, this means that even in the best circumstances, 5% to 10% of free flaps and replants fail. Almost all failures are due to vessel thrombosis, resulting in ischemia of the transplanted tissue. Many attemps have been undertaken to treat and reverse its effects. Zdeblick and colleagues noted an improvement in the viability of amputated limbs replanted after an extended period of ischemia following intraarterial infusion of urokinase. Subsequent studies have investigated many modalities of urokinase administration in various animal models by differing ischemic periods. These studies, however, have failed to establish a definitive, generally accepted protocol for administration of urokinase in the salvage of tissue subjected to prolonged ischemia. Our clinical observations suggest that a bolus of urokinase delivered under pressure may increase the thromoblytic effect of the drug, probably by means of increased delivery to microvasculature. We intend to investigate the role of selective pressure perfusion of ischemic flaps as a new means for increasing the effectiveness of urokinase in the treatment of the "no-reflow" phenomenon. A total of 32 male New Zealand rabbits were used and divided into the four groups according to the method of infusion. After 12 hours of ischemia the flaps were injected with Hartmann's solution or with urokinase and the percent survival of the flap was determined at 7 days following flap reperfusion. As the result, the flap survival rate was highest in the pressure injection of urokinase group.

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A comparative study of three therapeutic modalities in loculated tuberculous pleural effusions (소방이 형성된 결핵성 흉막 질환에서 배액 방법에 따른 치료효과)

  • Lee, Sang Hwa;Lee, So Ra;Lee, Sang Youb;Park, Sang Muyn;Suh, Jung Kyung;Cho, Jae Youn;Shim, Jae Jeong;In, Kwang Ho;Yoo, Se Hwa;Kang, Kyung Ho
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.5
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    • pp.683-692
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    • 1996
  • Background: Although most of the patients with tuberculous pleural effusions completely reabsorbed their effusions and became asymptomatic within 2 to 4 months, later surgical procedures such as decortication is needed in some patients because of dyspnea caused by pleural loculations and thickening despite anti-tuberculous chemotherapy. It is obligatory to secure adequate drainage to prevent the development of complications. But, the best methods for treating loculated tuberculous pleural effusions remain debatable. Recent several reports revealed that intrapleural instillation of fibrinolytic agents is an effective adjunct in the management of complicated empyema and may reduce the need of surgery. Purpose : The effects of catheterization with intrapleural urokinase instillation were prospectively evaluated in the patients with septated tuberculous pleural effusion, and compared with other therapeutic effects of different modalities of therapy such as repeated thoracentesis and small-bored catheterization. Methods : Forty-eight patients diagnosed with tuberculous pleurisy were randomly separated into three groups; control group(n=13), catheter group(n=12), urokinase group(n=22). In urokinase group, dose of 100.000U urokinase was instilled into the pleural cavity via a percutaneous drainage catheter for complete drainage or total dose of 700,000U of urokinase. After two hours clamping, the catheter was opened and intermittently irrigated. The early and late effectiveness of therapies was assessed by radiographically and by measuring the volume of fluid drained from the catheter. Results : There was statistically significantly better result in the urokinase group in respect of frequency of catheterization, frequency of catheter obstruction and the duration of catheterization in early effectiveness(p < 0.05). There were no difference in radiologic improvement of follow-up in later phase chest X-ray between urokinase group and catheter group in later phase(p > 0.05). But there were more failure rates in control group especially honeycomb septa in pleural effusion sonographically than former two groups. And there were no complications of urokinase such as fever or hemorrhage. Conclusion : In the treatment of septated tuberculous pleurisy, there were better results in urokinase than those of catheterization alone in early effectiveness. And there was no difference in radiographic improvement between urokinase group and catheter group. Intrapleural instillation of urokinase is an effective and safe mode of treatment for septated tuberculous pleural effusions and alleviates the need for thoracotomy.

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Intraarterial Catheter-directed Urokinase Infusion for Femoral Artery Thrombosis after Cardiac Catheterization in Infants and Children (심도자술후 발생한 대퇴동맥 혈전증 환아에서 동맥내 Urokinase 국소 주입요법의 효과)

  • Lee, Hyoung Doo
    • Clinical and Experimental Pediatrics
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    • v.45 no.11
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    • pp.1397-1402
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    • 2002
  • Purpose : One of the major complication of arterial catheterization is the thrombosis of the iliac or femoral arteries. Tissue loss following femoral artery catheterization is rare. However longterm sequelae such as impaired limb growth and future impairment of vascular access, are also important in pediatric cardiac patients. But standard methods to treat thrombotic complication of arterial catheterization in infants and children is not established. The present study was performed to assess the efficacy of intraarterial catheter-directed urokinase infusion in infants and children with limb ischemia due to arterial thrombosis after cardiac catheterization. Methods : From January 1994 to August 2002, 12 patients with thrombotic femoral artery occlusion after arterial catheterization were treated with catheter-directed urokinase infusion in Dong-A University Hospital. Retrospective analysis of the medical records and angiograms was conducted. Results : The incidence of femoral artery thrombosis after retrograde arterial catheterization, which had not responded to systemic infusion of heparin and/or urokinase, was 2.8 percent. The doses of urokinase were 1,000-4,400 unit/kg/hr and duration of infusion was $50.6{\pm}29.2$ hours(18-110 hours). Clot resolution was complete in all patients who started to receive the intraarterial urokinase infusion within four days after catheterization. Only partial thrombolysis was seen in two patients who were treated with intraarterial urokinase on the 12th and 19th days after thrombus formation. Balloon angioplasty was done for these two patients with partial success. Bleeding complications were seen in two cases. Conclusion : Early use of catheter-directed intraarterial infusion of urokinase is safe and effective in thrombolysis of femoral artery occlusion after cardiac catheterization in infants and children.

Fibrinolysis with Lower Dose Urokinase in Patients with Complicated Parapneumonic Effusion

  • Lee, Seul;Lee, Heock;Lee, Dong Hyun;Kang, Bo Hyoung;Roh, Mee Sook;Son, Choohee;Kim, Sung Hyun;Lee, Hyun-Kyung;Um, Soo-Jung
    • Tuberculosis and Respiratory Diseases
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    • v.84 no.2
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    • pp.134-139
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    • 2021
  • Background: Intrapleural urokinase is one of the most widely used fibrinolytic agents in the treatment of complicated parapneumonic effusion (CPPE). However, little research has been performed on the optimal urokinase dosage. The aim of this study was to evaluate the treatment efficacy of half dose urokinase compared with conventional dose urokinase. Methods: We retrospectively enrolled 92 patients with CPPE or empyema who underwent intrapleural urokinase treatment at two tertiary hospitals. Patients received antibiotics, chest tube drainage, and other treatments as part of routine care. The primary outcome was the treatment success rate in the half dose urokinase group (50,000 IU daily for maximal 6 days) and the conventional dose urokinase group (100,000 IU daily). Treatment success was defined as clinical and radiological improvements without surgical treatment or re-admission within one month. Results: Forty-four patients received half dose urokinase, whereas 48 patients were treated with conventional dose urokinase. Both groups were relatively well matched at baseline, excluding higher serum white blood cell count and higher empyema prevalence in the half dose urokinase group. The treatment success rate was not different between the two groups (p=0.048). There were no differences in the rate of in-hospital death and surgical treatment, hospitalization duration, and indwelling catheter duration. In the multivariate analysis, urokinase dose was not a predictor of treatment success. Conclusion: Half dose intrapleural urokinase is equally effective conventional dose urokinase in treating patients with CPPE or empyema.

Urokinase Conjugated with Water-Soluble Dextran

  • Kim Nam Deuk;Kim Hyun-Pyo;Byun Si Myung;Kim Sung Wan
    • Bulletin of the Korean Chemical Society
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    • v.6 no.4
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    • pp.210-214
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    • 1985
  • Urokinase, a plasminogen activator, was conjugated with dextran by the cyanogen bromide activation-coupling method. The resulting water-soluble conjugate was purified by gel permeation chromatography on Sephadex G-200. The maximal activity was obtained when the ratio of urokinase/dextran was 1/20 for the coupling. The final preparation showed 5 CTA units/mg conjugate, 300 CTA units/mg protein, 8.4 % activity retention, and 47 % protein retention. The urokinase-dextran conjugate had good thermal, pH and storage stabilities. In addition, it showed greater resistance to the inhibitory effect of human plasma than native urokinase. Also in vitro biological half-life of urokinase increased 40 times by this conjugation. In view of activity, excellent stability and increased half-life, the conjugate can be a potential fibrinolytic agent in an injectable form.

Scale-up of Covalently Immobilized Urokinase Column and Repeated Use of It by Solid-Phase Refolding (공유결합으로 고정화된 urokinase 칼럼의 스케일업과 solid-phase refolding에 의한 반복 사용)

  • 서창우;최강선;이은규
    • KSBB Journal
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    • v.16 no.5
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    • pp.500-504
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    • 2001
  • We scaled up a covalent immobilization system of urokinase to the activated Sepharose and used it repeatedly to cleava a fusion protein consisting of human growth hormone and GST fragment. After scale up from 6 ml to 250 ml. the column system still demonstrated basically the same performance in terms of urokinase immobilization and fusion protein cleavage. When the column was washed with 6 M guanidine HCI after the cleavage reaction, the immobilized urokinase showed no activity probably becasue it was fully unfoled. However, as the denaturant was gradually removed from the column the immobilized urokinase fully regained its bioactivity, which indicated it was properly refolded into is natie conformation as covalently attached to the solid matrix. After 20 cycles of this solid-phase unfolding/refolding. the immobilized urokinase maintained approx. 80% of the initial bioactivity. This method provides and efficient protocol to apply the solid-phase refolding technique to improve the longevity of immobilized enzyme columns.

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Protective Effect of Urokinase on Reperfusion Function in Isolated Perfused Rat Heart, and Anti-platelet Aggregation Effect Invitro and Exvivo (Urokinase의 적출심장의 심근허혈에 대한 보호작용과 invitro 및 exvivo항혈전작용 실험)

  • Kwon, Kwangil;Shin, Hongseup;Yoon, Jongok;Kim, Boshin;Min, Jiha;Lee, Byungho;Huh, Inhoe
    • Korean Journal of Clinical Pharmacy
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    • v.2 no.1
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    • pp.1-9
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    • 1992
  • Protective effect of urokinase on reperfusion were studied followed by global ischemia in the isolated perfused rat heart. Separately, anti-platelet aggregation effect of urokinase also investigated. Urokinase exhibited positive effect for the protection of rat heart function by increasing the LV dp/dt, coronary flow(CF) and the Tate pressure product(RPP), and by decreasing the LVEDP on reperfusion. Urokinase also decreased arrhythmia by $74.7\%(P<0.05) induced by global ischemia in the rat heart. In the platelet aggregation study, urokinase did not show the inhibitory effect of ADP or collagen induced platelet aggregation inviuo and exvivo.

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Burr hole drainage using urokinase for treatment of subacute subdural hematoma (아급성기 경막하 혈종 치료시 천공술 후 urokinase를 이용한 혈종배액술)

  • Kim, Min-Su;Kim, Seong-Ho;Kim, Oh-Lyong
    • Journal of Yeungnam Medical Science
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    • v.32 no.1
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    • pp.8-12
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    • 2015
  • Background: Enlargement of subdural hematomas is relatively rapid in subacute stage of hematoma with clinical deterioration, which eventually necessitates surgery. The purpose of this study is to investigate the feasibility and safety of burr hole drainage using urokinase for management of patients with subacute subdural hematoma (SASDH). Methods: Nine patients with SASDH were treated by burr hole drainage using urokinase. Under local anesthesia a catheter was inserted into the hematoma through a burr hole. Burr hole drainage was followed by hematoma thrombolysis with instillation of urokinase (10,000 units) every 12 hours. Drainage was discontinued when a significant decrease of hematoma was observed on cranial computed tomography. Results: The patients' median age was 70 years (range, 62-87). The median Glasgow Coma Scale score before surgery was 15 (range, 11-15). Drainage was successfully performed in all patients. All patients had Glasgow Outcome Scale scores of 5 at discharge. There was no surgery-related morbidity or mortality. Conclusion: A burr hole drainage using urokinase could be a safe, feasible and effective minimally invasive method with low morbidity in treatment of selected patients with SASDHs.