• Title/Summary/Keyword: Fibrinolysis

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Induction of Apoptosis by Ethanol Extracts of Fermented Agabeans in AGS Human Gastric Carcinoma Cells (AGS 인체위암세포에서 발효된 아가콩 추출물에 의한 apoptosis 유도)

  • Kim, Sung-Ryeal;Lee, Hye-Hyeon;Kim, Min-Jeong;Seo, Min-Jeong;Hong, Su-Hyun;Choi, Yung-Hyun;Kang, Byoung-Won;Park, Jeong-Uck;Joo, Woo-Hong;Rhu, Eun-Ju;Jeong, Yong-Kee
    • Journal of Life Science
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    • v.20 no.12
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    • pp.1872-1881
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    • 2010
  • Extracts of soybeans fermented by Bacillus subtilis have a wide variety of functions, such as enhancing the body's immune function, fibrinolysis activity, anti-inflammation, anti-cancer, estrogen function and anti-infection effects. Recently, it was reported that the extracts of fermented beans exhibit strong anti-inflammatory and anti-cancer properties by suppressing the transcription of pro-inflammatory cytokine genes and induction of apoptosis, respectively. However, the mechanisms of their cytotoxicity in human gastric cancer cells are poorly understood. In the present study, we investigated the effects of ethyl alcohol extracts from fermented soybean (FS) and yellow agabean (FYA) on cell growth and apoptosis in AGS human gastric cancer cells. A treatment of FS and FYA inhibited the growth of AGS cells in a concentration-dependent manner by inducing apoptosis. FS- and FYA-induced apoptosis were associated with down-regulation of XIAP and cIAP-2, and up-regulation of pro-apoptotic Bax expression. Moreover, a treatment of FS and FYA not only triggered an increase in the levels of death receptor (DR)4, DR5, Fas and FasL, but also induced the activation of casepase-3, -8 and -9. These findings illustrate that FS and FYA may have a therapeutic potential in human gastric AGS cells and as a functional food.

Effects of Cumulus Cells and Reactive Oxygen Species (ROS) on Plasminogen Activator Activity during In Vitro Maturation of Porcine Oocytes

  • Sa, Soo-Jin;Park, Chun-Keun;Kim, In-Cheul;Lee, Seung-Hoon;Kwon, Oh-Sub;Kim, Myung-Jick;Cho, Kyu-Ho;Kim, Du-Wan;So, Kyoung-Min;Cheong, Hee-Tae;Webb, Bob
    • Journal of Embryo Transfer
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    • v.25 no.3
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    • pp.171-177
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    • 2010
  • Plasminogen activators (PAs) are serine proteases that convert plasminogen to plasmin. The PA/plasmin system has been associated with a number of physiological processes such as fibrinolysis, ovulation and fertilization. Although correlations have been reported between reactive oxygen species (ROS) and oocyte maturation, the relationship between PA activity and ROS is unknown. The present study was undertaken to determine the effects of cumulus cells on PA activity in matured porcine oocytes under xanthine (X)-xanthine oxidase (XO) system. When oocytes were matured under the X-XO system, the proportion of oocytes remaining GV stage was higher (p<0.05) in oocytes without cumulus cells. The incidence of degenerated oocytes was higher (p<0.05) in the X+XO ($11.1{\pm}6.1$ and $21.6{\pm}3.4%$) than in the control group ($2.9{\pm}1.8$ and $4.0{\pm}1.6%$). The proportion of TUNEL-positive oocytes and activity of caspase-3 were higher (p<0.05) in cumulus-free oocytes and oocytes exposed to ROS. Tissue-type plasminogen activator-plasminogen activator inhibitor (tPA-PAI) and tissue-type plasminogen activator (tPA) activity were detected in oocytes that were separated from cumulus-oocytes complexs (COCs) at 44 h of maturation culture, and only tPA was produced in oocytes that were denuded before the onset of maturation culture. On the other hand, the activities of PA were increased (p<0.05) when oocytes were cultured under the X-XO system. The higher activity of tPA was observed in denuded oocytes (DOs) underwent apoptotic changes by oxidative stress. In COCs, however, tPA-PAI as well as tPA activity was detected and apoptotic changes such as DNA cleavage or caspase-3 activation were not observed. These results suggest that tP A may be relevant to apoptotic cell death in porcine oocytes by oxidative stress.

FIBRIN SEALANTS IN MAXILLOFACIAL SURGERY : A INTRODUCTORY REPORT (악안면 외과 영역에서의 FIBRIN SEALANTS 의 이용)

  • Kim, Myung-Jin;Park, Hyung-Kook
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.13 no.2
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    • pp.129-136
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    • 1991
  • The fibrin sealant was first designed as an alternative to surgical suture for the purpose of surface-to-surface union especially in parenchymal organs like the liver, spleen and kidney. The clinical application of currently used fibrin sealant was first introduced in 1972. The fibrin sealant consists of principal two components; lyophilized human fibrinogen and bovine thrombin. The fibrinogen component also contains coagulation factor XIII. A solution of aprotinin, an inhibitor of fibrinolysis is used to dissolve the fibrinogen and to provide the first component, and a solution of calcium chloride is also used to provide the second component. From July to December in 1990, during 6 months, we used fibrin sealant in the 28 patients of 33 various cases, in the following ways; supportive application of fibrin sealant after free autogenouse nerve graft for the repair of inferior alveolar nerve, facial nerve or accessory nerve, treament of hemangioma or lymphangioma to thrombosize and lead to the tumor shrinking, skin grafting to stimulate the adhesion and tissue repair, bone grafting in the patients of cleft alveolus, mandibular reconstruction or orthognathic surgery to facilitate the knitting of bone chips, tissue adhesion after tumor resection, radical neck dissection or flap reconstructions, and supportive adhesion of external auditory cannal after TMJ surgery via postauricular approach. No adverse effects were observed, none of the patients developed hepatitis or other blood transmitted disease, and the wound healing were acceptable.

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Evaluation of Plasma D-Dimer Concentration in Dogs with Chronic Mitral Valve Insufficiency (만성 이첨판 폐쇄부전증에 걸린 개에서 혈장 D-dimer 농도 측정 연구)

  • Park, Joungsoon;Suh, Sang-Il;Oh, Yeonsu;Hyun, Changbaig
    • Journal of Veterinary Clinics
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    • v.32 no.1
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    • pp.5-8
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    • 2015
  • D-dimer is a fibrin degradation product (FDP), a small protein fragment present in the blood after a blood clot is degraded by fibrinolysis. D-dimer concentration is widely used for determining thrombosis and thromboembolism. Because one major cause of thromboembolism is congestive heart failure in human, we investigated the degree and risk of thromboembolism in dogs with different stage of congestive heart failure caused by chronic mitral valvular insufficiency (CMVI). The plasma level of d-dimer was evaluated in 20 healthy dogs and 30 dogs with different stage of congestive heart failure caused by CMVI. The d-dimer concentrations were measured by a commercialized assay kit. The plasma levels of d-dimer were not significantly different between healthy and CMVI dogs. Furthermore, there was no association of d-dimer concentrations to left atrium to aorta (LA/Ao) ratio, left ventricular dimension at diastole to aorta (LVIDd/Ao) ratio and severity of heart failure in our study population. Our study results implied that the degree of thromboembolism in canine heart failure might be minimal or the plasma d-dimer test might not be reliable for detecting thromboembolism in dogs.

Experimental Endotoxin-Induced Disseminated Intravascular Coagulation in Rat Model (쥐 모델에 있어 내독소에 의한 실험적인 범발성 혈관내 응고증)

  • Seok- Cheol Choi;Jai-Young Kim;Jin-Bog Koh;Won-Jae Lee
    • Biomedical Science Letters
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    • v.3 no.2
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    • pp.83-88
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    • 1997
  • In septic patients, disseminated intravascula. coagulation (DIC) occurs frequently and is a pathologic condition associated with a variety of critical illness. DIC may complicate the already complex clinical situations and contribute to the high mortality. Nevertheless, its pathogenic mechanisms are not completely elucidated. Present study was prospectively designed to understand the pathogenetic mechanisms involved in the development of DIC. 15 rats were subjected to study and according to the aim, they were divided into three groups: group I, control (not treated-endotoxin, n=5); group II (12 hours after endotoxin injection, n=5); group III (24 hours after endotoxin injection, n=5). Experimental DIC was induced in rats by a bolus injection of endotoxin (1mg/kg, E. coli serotype 055:B5). Blood was collected by direct puncture of the heart. Platelet count, fibrinogen and plasminogen concentration, antithrombin III, D-dimer and complement components (C3 and C4) were measured in all subjects. In group II and III, there were apparent signs of DIC, including thrombocytopenia, decreased fibrinogen (but increase in group III), reduced C3 and antithrombin III, and elevated D-dimer. These data indicated that endotoxin might induce the activation of several pathways such as coagulation, fibrinolytic and complement cascade, causing DIC and subsequent multiple organ failures. Ultimately, the increased knowledge of the various pathogenetic mechanisms of coagulation activation and fibrinolysis in endotoxin-induced DIC may have prophylactic or therapeutic implications.

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Meaning of the DR-$70^{TM}$ Immunoassay for Patients with the Malignant Tumor (악성 종양 환자에 대한 DR-$70^{TM}$ 면역 분석법의 의의: Validation Study)

  • Lee, Ki-Ho;Cho, Dong-Hee;Kim, Sang-Man;Lee, Duck-Joo;Kim, Kwang-Min
    • IMMUNE NETWORK
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    • v.6 no.1
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    • pp.43-51
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    • 2006
  • Background: The DR-$70^{TM}$ immunoassay is a newly developed cancer diagnostic test which quantifies the serum fibrin degradation products (FDP), produced during fibrinolysis, by antibody reaction. The purpose of this study was to evaluate the potential of DR-$70^{TM}$ Immunoassay in screening malignant tumor. Methods: Sample subjects were 4,169 adults, both male and female, who visited the health promotion center of a general hospital from March 2004 to April 2005 and underwent the DR-$70^{TM}$ immunoassay test and other tests for cancer diagnosis. The patient group was defined as 42 adults out of the sample subjects who were newly diagnosed with cancer during the same time period when the DR-$70^{TM}$ immunoassay test was performed. Final confirmation of a malignant tumor was made by pathological analysis. Results: The mean DR-$70^{TM}$ level was $0.83{\pm}0.65{\mu}g/ml$ (range: 0.00 (0.0001)${\sim}7.42{\mu}g/ml)$ in the control group (n=4,127) as opposed to $2.70{\pm}2.33{\mu}g/ml$ (range: $0.12{\sim}9.30{\mu}g/ml)$ in the cancer group (n=42), and statistical significance was established (p<0.0001, Student t-test). When categorized by the type of malignant tumor, all cancer patients with the exception of the subgroups of colon and rectal cancer showed significantly higher mean DR-$70^{TM}$ levels compared with the control group (p<0.0001, Kruscal-Wallis test). The receiver operating characteristic (ROC) curve analysis revealed ${\geq}1.091{\mu}g/ml$ as the best cut-off value. Using this cut-off value, the DR-$70^{TM}$ immunoassay produced a sensitivity of 71.4%, a specificity of 70.1%, a positive predictability of 69.4%, and a negative predictability of 69.2% (1). Conclusion: A significant increase in the mean DR-$70^{TM}$ value was observed in the cancer group (thyroidal, gastric, breast, hepatic and ovarian) com pared with the control group. In particular, the specificity and sensitivity of the DR-$70^{TM}$ immunoassay was relatively high in the subgroups of breast, gastric, and thyroidal cancer patients. There is need for further studies on a large number of malignant tumor patients to see how the DR-$70^{TM}$ level might be changed according to the differentiation grade and postoperative prognosis of the malignant tumor.

Isolation and Physiological Characteristics of Microorganisms Producing Extracellular Enzymes from Korean Traditional Soybean Sauce and Soybean Paste (전통 장류에서 세포외효소 분비능이 우수한 미생물의 분리 및 생리활성 특성)

  • Baek, Seong-Yeol;Yun, Hye-Ju;Choi, Hye-Sun;Koo, Bon-Sung;Yeo, Soo-Hwan
    • Microbiology and Biotechnology Letters
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    • v.38 no.4
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    • pp.379-384
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    • 2010
  • We isolated microorganisms presenting high enzymatic activities for amylase, cellulase, protease, lipase or fibrinolysis from Korean traditional soybean sauce and paste. Then, the physiological properties and 16S rRNA sequences of isolated microorganisms were analyzed. All of the isolated 13 strains possessing high extra cellular enzyme activities have higher amylase and cellulase activities than Bacillus subtilis KACC 10114. All the selected strains have protease activities except for D2-14. Except D8-8 and K4-1, other strains have lipase activity. D2-7, D8-8 and K4-1 strains have higher fibrinolytic activities than others, while D8-2 strain has no activity. Most of the selected strains showed antibacterial activity even in gram positive and gram negative bacteria and yeast. Gene sequence analysis of 16S rRNA from isolated strains revealed that all the selected strains were member of Bacillus species.

Bacillus subtilis를 이용한 대두 발효식품의 혈전용해능

  • Jeong, Yeong-Gi
    • Proceedings of the Korean Society of Life Science Conference
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    • 2001.06a
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    • pp.67-86
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    • 2001
  • A strain producing strongly fibrinolytic enzyme was isolated from soil and was identified to be Bacillus subtilis by biochemical and physiological characterization. The optimal culture conditions for the production of fibrinolytic enzyme was determined to be 1.0% tryptone, 1.5% soluble starch, 0.5% Peptone, 0.5% NaCl, $(NH_{4})_{3}PO_4.3H_{2}O, and MgSO_{4}.7H_{2}O.$ Initial pH and temperature were pH 8.0 and $30^{\circ}C$ , respectively, The highest enzyme production was observed at 30 hours of cultivation at $30^{\circ}C$ The fibrinolytic enzyme was purified to homogeneity by DEAE Sephadex A-50 ion exchange column chromatography, 70% ammonium sulfate precipitation, Sephadex G-200 and G-75 gel filtration column chromatography. The molecular weight of the purified enzyme was 28,000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A gene encoding the fibrinolytic enzyme was cloned into a plasmid vector pBluescript, transforming E.coli XL-1 Blue. The clone was able to degrade fibrin, This indicated that the gene could encode a fibrinolytic enzyme. The nucleotide sequence of the 2.7 kb insert was determined in both direction. One open reading frame composed of 1023 nucleotides was found to be a potential protein coding region. There was the putative Shine-Dalgano sequence and TATA box upstream of the open reading frame. The homology search data in the genome database showed that both the 2.7 kb insert and 1 kb open reading frame carried no significance in the nucleotide sequence of known fibrinolytic enzyme from Bacillus serovars. The recombinant cell harboring the novel gene involved in fibrinolysis was subjected to protein purification. The molecular mass of the purified fibrinolytic enzyme was determined to be 31864 Dalton, which was highly in accordance with the molecular mass(33 kDa) of the fibrinolytic gene deduced from the insert. The fibrinolytic enzyme was Purified 50.5 folds to homogeneity in overall yield of 10.7% by DEAE Sephadex A-50 ion exchange, 85% ammonium sulfate precipitation, Sephadex G-50, Superdex 75 HR FPLC gel filtration. In conclusion, a novel fibrinolytic gene from Bacillus subtilis was identified and characterized by cloning a genomic library of Bacillus subtilis into pBleuscript. For the soybean fermented by this strain, it is found that there increased assistant protein about 20% compared to the soybean not fermented and increased about 30% according to amino acid analysis and, in particular, essential amino acid increased about 40%. When keeping this fermented soybean powder at room temperature for about 70days, it showed very high stability maintaining almost perfect activity and, therefore, it gave us great suggestion its possibility of development as a new functional food.

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Polymorphisms of 3-hydroxy-3-methylglutaryl Coenzyme A Reductase Gene Are Not Associated with the Osteonecrosis of Femoral Head in Korean (한국인에서 HMG-CoA reductase 유전자다형성과 대퇴골두무혈성괴사증과의 연관성 분석)

  • Kim, Tae-Ho;Hong, Jung-Min;Lee, Sang-Han;Park, Eui-Kyun;Kim, Shin-Yoon
    • Journal of Life Science
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    • v.18 no.4
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    • pp.427-434
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    • 2008
  • Osteonecrosis of the femoral head (ONFH) is a multifactorial disease and certain individuals are more at risk or may be predisposed to it. An altered lipid metabolism is one of the major risk factors for osteonecrosis, especially corticosteroid therapy and alcoholism. 3-hydroxy-3-methylglutaryl coenzyme A. (HMG-CoA) reductase inhibitors, stalin used as lipid-clearing agent, have been known to decrease the risk of osteonecrosis in patients receiving steroids and affect coagulation and fibrinolysis. Therefore we evaluated the association of HMG-CoA reductase gene polymorphisms and haplotypes between osteonecrosis patients and normal controls. We directly sequenced the HMG-CoA reductase gene in 24 Korean individuals, and identified five sequence variants. Four SNPs (-6933C>T, -6045T>G, +12673G>A, and +18128C>T) were selected and genotyped in 349 male ONFH patients and 300 male control subjects. The genotypes, allele frequencies, and haplotypes of the polymorphisms in the total patients as well as in the subgroup by etiology were not significantly different from those in the control group. In addition, no significant differences between each genotype of the polymorphisms and plasma lipid level could be found in the control group. These results suggest that the polymorphisms and haplotypes of HMG-CoA reductase gene are unlikely to be associated with a susceptibility to ONFH.

Hematologic Changes and Factors Related to Postoperative Hemorrhage Following Cardiopulmonary Bypass (체외순환에 따른 혈액학적 변화와 술후 출혈에 관계하는 인자에 관한 연구)

  • 김하늘루;황윤호;최석철;최국렬;김승우;조광현
    • Journal of Chest Surgery
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    • v.31 no.10
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    • pp.952-963
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    • 1998
  • Background: Cardiopulmonary bypass(CPB)-induced hemostatic defects may result increased possibility of excessive hemorrhage and additional multiple transfusion reactions or reoperation. Particularly, fibrinolytic activation and decreased platelet count and function by CPB were proposed as a predictor of hemorrhage during postoperative periods in several reports. Materials and methods: Present study, which was conducted in 20 adult patients undergoing CPB, was prospectively designed to examine the hematologic changes, including fibrinolytic activation during and after CPB and to clarify the relationships between these changes and the magnitude of the postoperative nonsurgical blood loss. The serial blood samples for measurment of hematologic parameters were taken during operation and postoperative periods. Blood loss was respectively counted via thoracic catheter drainage at postoperative 3, 6, 12, 24, 48 hours and total period. Results: The results were obtained as follows:Platelet count rapidly declined following CPB(p<0.01), which its decreasing rate was an inverse proportion to total bypass time(TBT, r=0.55, p=0.01), And platelet count in postoperative 7th day was barely near to its control value. Fibrinogen degradating product(FDP) and D-dimer level significantly increased during CPB(p<0.0001, p<0.0001, respectively), and both of fibrinogen and plasminogen concentration correlatively decreased during CPB(r=0.57, p<0.01), implying activation of fibrinolytic system. Postoperative bleeding time (BT), postoperative activated partial thromboplastin time(aPTT) and postoperative prothrombin time (PT) were significantly prolonged as compare with each control value (p=0.05, p<0.0001, p<0.0001, respectively). Total blood loss was positively correlated with patient's age, aortic clamping time (ACT) and TBT, while there was negative correlation between platelet count and blood loss at pre-CPB, CPB-off and the 1st postoperative day, and in some periods. Postoperative aPTT and postoperative PTwere positively related to postoperative 6 hr and 48 hr blood loss(r=0.53, p=0.02; r=0.43, p=0.05) but not to total blood loss, whereas there was no relationship between postoperative BT and blood loss at any period. Conclusions: These observations suggest that CPB results various hematologic changes, including fibrinolytic activation and severe reduction in platelet count. Diverse factors such as age, platelet count, ACT, TBT and postoperative aPTT and PT may magnify the postoperative bleeding. This study will be a basic reference in understanding CPB-induced hemostatic injuries and in decreasing the postoperative hemorrhage

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