• Journal of Korean Neurosurgical Society
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• 제66권3호
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• pp.263-273
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• 2023

While the survival rate of preterm infants has increased dramatically over the last few decades, intraventricular hemorrhage and subsequent hydrocephalus remain major unsolved problems in neonatal intensive care. Once intraventricular hemorrhage occurs, severe neurological sequelae are inevitable. Treatment of this complicated pathology and achievement of favorable neurofunctional outcomes in fragile infants are crucial challenges for pediatric neurosurgeons. Fibrinolytic therapy, which chemically dissolves hematoma, is a promising and useful treatment method. In this paper, the historical background of fibrinolytic therapy for post-intraventricular hemorrhagic hydrocephalus in preterm infants is reviewed and a recent method of fibrinolytic therapy using urokinase is introduced.

• The Journal of Korean Physical Therapy
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• 제25권4호
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• pp.224-231
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• 2013

Purpose: The purpose of this case study was to investigate three poor fibrinolytic responders with chronic ischemic stroke to acute exercise intensity and time. Methods: Three ischemic stroke patients (male) from the stroke center located at Busan metropolitan area in Republic of Korea volunteered at this study. They performed two single session exercises that were a VO2peak test and a single bout treadmill walking (70-75%HRpeak, 30 min, 50min). Fasting blood samples for determination of tissue Plasminogen Activator (tPA) and Plasminogen Activator Inhibitor-1 (PAI-1) were obtained before, immediately after, 30min after acute exercise. SPSS 12.0 was used for analyzing of data and computing mean and standard deviation, and change rate was conducted between times. Results: In fibrinolytic activity according to the intensity and time of acute exercise, tPA change increased steadily during the recovery stage after the VO2peak in the cases, but PAI-1 activity showed different patterns among the cases. In a single bout treadmill walking (70-75%HRpeak, 30 min, 50min), tPA change increased between 30min and 50min. Conclusion: In conclusion, these results suggest that the exercise prescription for poor fibrinolytic responder with three male chronic ischemic stroke patients without motor disability recommend at 70-75%HRpeak, over 30min.

• Tuberculosis and Respiratory Diseases
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• 제76권3호
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• pp.127-130
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• 2014

The risk of dying from a pulmonary embolism (PE) is estimated to be about 30% if inotropic support is required and no cardiopulmonary arrest occurs. Fibrinolysis in massive PE is regarded as a life-saving intervention, unless there is a high risk of bleeding following the use of the fibrinolytic therapy. Rivaroxaban is an oral factor Xa inhibitor, however its anticoagulation effects before or after administration of fibrinolytics in massive PE are still unknown. Two patents were admitted: 61-year-old woman with repeated syncope, and a 73-year-old woman was admitted with dyspnea and poor oral intake. Systemic arterial hypotension with radiologic confirmation led to a diagnosis of massive PE in both patients. Rivaroxaban was administered before in one, and after firbrinolytic therapy in the other. One showed similar efficacy of rivaroxaban with currently used anticoagulants after successful fibrinolysis, and the other one without antecedent administration of the fibrinolytic agent showed unfavorable efficacy of rivaroxaban.

• 대한의용생체공학회:의공학회지
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• 제15권1호
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• pp.9-18
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• 1994

• Journal of Microbiology and Biotechnology
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• 제25권9호
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• pp.1449-1459
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• 2015

A novel proteolytic enzyme with fibrinolytic activity, FSP3, was purified from the recently isolated Streptomyces sp. P3, which is a novel bacterial strain isolated from soil. FSP3 was purified to electrophoretic homogeneity by ammonium sulfate precipitation, anion exchange, and gel filtration. FSP3 is considered to be a single peptide chain with a molecular mass of 44 kDa. The maximum activity of the enzyme was observed at 50℃ and pH 6.5, and the enzyme was stable between pH 6 and 8 and below 40℃. In a fibrin plate assay, FSP3 showed more potent fibrinolytic activity than urokinase, which is a clinical thrombolytic agent acting as a plasminogen activitor. The activity was strongly inhibited by the serine protease inhibitor PMSF, indicating that it is a serine protease. Additionally, metal ions showed different effects on the activity. It was significantly suppressed by Mg²⁺ and Ca²⁺ and completely inhibited by Cu²⁺, but slightly enhanced by Fe²⁺. According to LC-MS/MS results, its partial amino acid sequences are significantly dissimilar from those of previously reported fibrinolytic enzymes. The sequence of a DNA fragment encoding FSP3 contained an open reading frame of 1287 base pairs encoding 428 amino acids. FSP3 is a bifunctional enzyme in nature. It hydrolyzes the fibrin directly and activates plasminogen, which may reduce the occurrence of side effects. These results suggest that FSP3 is a novel serine protease with potential applications in thrombolytic therapy.

• Journal of Microbiology and Biotechnology
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• 제31권2호
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• pp.327-337
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• 2021

Fibrinolytic enzymes with a direct mechanism of action and safer properties are currently requested for thrombolytic therapy. This paper reports on a new enzyme capable of degrading blood clots directly without impairing blood coagulation. This enzyme is also non-cytotoxic and constitutes an alternative to other thrombolytic enzymes known to cause undesired side effects. Twenty-four Bacillus isolates were screened for production of fibrinolytic enzymes using a fibrin agar plate. Based on produced activity, isolate S127e was selected and identified as B. subtilis using the 16S rDNA gene sequence. This strain is of biotechnological interest for producing high fibrinolytic yield and consequently has potential in the industrial field. The purified fibrinolytic enzyme has a molecular mass of 27.3 kDa, a predicted pI of 6.6, and a maximal affinity for Ala-Ala-Pro-Phe. This enzyme was almost completely inhibited by chymostatin with optimal activity at 48℃ and pH 7. Specific subtilisin features were found in the gene sequence, indicating that this enzyme belongs to the BPN group of the S8 subtilisin family and was assigned as AprE127. This subtilisin increased thromboplastin time by 3.7% (37.6 to 39 s) and prothrombin time by 3.2% (12.6 to 13 s), both within normal ranges. In a whole blood euglobulin assay, this enzyme did not impair coagulation but reduced lysis time significantly. Moreover, in an in vitro assay, AprE127 completely dissolved a thrombus of about 1 cc within 50 min and, in vivo, reduced a thrombus prompted in a rat tail by 11.4% in 24 h compared to non-treated animals.

• Journal of Microbiology and Biotechnology
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• 제24권2호
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• pp.245-253
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• 2014

A fibrinolytic enzyme was produced by an edible mushroom of Pleurotus ostreatus using submerged culture fermentation. The enzyme was purified from the culture supernatant by applying a combination of freeze-thaw treatment, ammonium sulfate precipitation, hydrophobic interaction, and gel filtration chromatographies. The enzyme was purified by a 147-fold, with a yield of 7.54%. The molecular masses of the enzyme an determined by gel filtration and SDS-PAGE were 13.6 and 18.2 kDa, respectively. The isoelectric point of the enzyme was 8.52. It hydrolyzed fibrinogen by cleaving the ${\alpha}$ and ${\beta}$ chains of fibrinogen followed by the ${\gamma}$ chains, and also activated plasminogen into plasmin. The enzyme was optimally active at $45^{\circ}C$ and pH 7.4. The enzyme activity was completely inhibited by EDTA, whereas protease inhibitors of TPCK, SBTI, PMSF, aprotinin and pepstatin showed no inhibition on its activity. The partial amino acid sequences of the enzyme as determined by Q-TOF2 were ATFVGCSATR, GGTLIHESSHFTR, and YTTWFGTFVTSR. These sequences showed a high degree of homology with those of metallo-endopeptidases from P. ostreatus and Armillaria mellea. The purified enzyme can also be applied as a natural agent for oral fibrinolytic therapy or prevention of thrombosis.

• Journal of Microbiology and Biotechnology
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• 제28권2호
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• pp.275-283
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• 2018

Ischemic stroke can result from blockage of blood vessels, forming fibrin clots in the body and causing irreparable brain damage. Remedial thrombolytic agents or anticoagulants have been studied; however, because the FDA-approved tissue plasminogen activator has low efficacy and side effects, it is necessary to develop safer and more effective treatment candidates. This study aimed at assessing the fibrinolytic and anticoagulation features of a novel serine protease extracted and purified from Diopatra sugokai, a polychaeta that inhabits tidal flats. The purified serine protease was obtained through ammonium sulfate precipitation, affinity chromatography, and ion-exchange chromatography. Its molecular size was identified via SDS-PAGE. To characterize its enzymatic activities, the protease activity at various pH and temperatures, and in the presence of various inhibitors, was measured via azocasein assay. Its fibrinolytic activity and anticoagulant effect were assessed by fibrin zymography, fibrin plate assay, and fibrinogenolytic activity assays. The novel 38 kDa serine protease had strong indirect thrombolytic activity rather than direct activity over broad pH (4-10) and temperature ($37^{\circ}C-70^{\circ}C$) ranges. In addition, the novel serine protease exhibited anticoagulant activity by degrading the ${\alpha}$-, ${\beta}$-, and ${\gamma}$-chains of fibrinogen. In addition, it did not produce cytotoxicity in endothelial cells. Therefore, this newly isolated serine protease is worthy of further investigation as a novel alkaline serine protease for thrombolytic therapy against brain ischemia.

• 한국버섯학회지
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• 제13권3호
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• pp.192-198
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• 2015

잎새버섯으로 준비된 물 추출물과 유기용매 분획물의 혈전용해활성과 트롬빈저해활성, acetylcholinesterase 저해활성, 항산화활성 및 면역활성을 확인하였다. 물 추출물과 물 분획물은 각각 1.55 plasmin unit와 0.85 plasmin unit의 높은 혈전용해활성을 나타내고, 물 분획물과 에틸 아세테이트 분획물은 76.43%와 72.59%의 트롬빈저해활성을 나타냈다. 클로로포름 분획물과 헥산 분획물이 95.14%와 94.74%의 높은 acetylcholinesterase 저해활성을 나타냈으며, 부탄올 분획물과 물 추출물이 94.47%와 91.04%의 항산화활성을 나타냈다. 부탄올 분획물과 물 분획물이 각각 2배 이상의 높은 면역기능활성을 나타냈다. 실험 결과로부터 잎새버섯의 분획물은 높은 혈전용해활성, 트롬빈 저해활성, acetylcholinesterase 저해활성 및 면역 기능활성을 나타내 심혈관계 질환과 치매의 예방 및 면역 기능강화를 위한 기능성식품 개발에 이용할 수 있을 것으로 사료된다.

• Tuberculosis and Respiratory Diseases
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• 제40권4호
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• pp.378-383
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• 1993

연구배경 : 최근의 연구에서는 염증성늑막삼출의 경우, 늑막내의 응고작용은 증가되고 섬 유소용해능은 감소되어 흉수의 섬유소 제거가 감소됨이 보고되었다. 특히 농흉의 경우에 있 어서 섬유소침착이 진행되어 늑막강내에 격막이 형성되는 경우를 섬유소화농기로 분류하는 데 이때는 임상적으로 다수의 소방이 상호 분리된 격막으로 형성됨으로 늑막천자로서는 배 농이 불가능하며 단순폐쇄식 흉부삽관술을 이용한 치료로서도 완치가 어렵다. 따라서 최근 의 늑막강내로의 유로키나제(UK) 국소적 주입이 좋은 성적을 보임으로써 국소적 섬유소용 해치료가 농흉치료의 효과적인 한 방편으로 제시되고 있다. 그러나 이러한 임상적 관찰에도 불구하고 UK주입후 과연 늑막강내의 섬유소용해능이 증가하는 지에 관한 직접적인 연구보 고는 없었다. 따라서 저자들은 늑막강내 UK의 주입이 국소적 섬유소용해능을 증가시켜 소 방의 용해를 촉진시킨다는 가설하에 농흉환자를 대상으로 유로키나제 주입전후의 흉수내 D-dimer(D-Di)와 plasminogen activator activity(PA-activity)를 측정하고자 하였다. 방법 : 다수의 소방이 형성된 14예의 농흉환자를 대상으로 UK주입전후의 흉수내 D-dimer를 효소결합면역흡착검사(ELISA)로 측정하였고 PA-activity는 광발색법으로 측정 하였다. 일회 주입시 20만단위의 UK를 사용하였고 모든 환자에서 최소 2회 이상 투여하였으며 3회 주입후 결과를 분석하였다. 결과 : UK 주입전 PA-activity는 $10.5{\pm}7.0$ IU tPA/ml로서 UK 주입 1회 후, 2회 후, 3회 후 각각 $91.9{\pm}27.0$, $432.3{\pm}177.1$, $170.0{\pm}85.3$ IU tPA/ml로 유의하게 상승하였고(p<0.01) 2회 째 주입후 최고치에 도달하였다(p<0.01). D-Di 역시 주입전 $4.16{\pm}1.06{\times}10^5ng/ml$에서 주입 1회 후, 2회 후 및 3회 후 각각 $9.62{\pm}1.54{\times}10^5$, $12.31{\pm}1.89{\times}10^5$, $8.54{\pm}1.56{\times}10^5ng/ml$으로 유의하게 상승하였다(p<0.05). 결론 : 늑막강내 유로키나제의 국소적 주입은 늑막내 plasmic을 생성하여 섬유소용해를 촉진시킴으로서 소방을 제거하는 것으로 보인다.