• Title/Summary/Keyword: Tissue Plasminogen Activator

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Studies of Attachment of Fibroblasts on Microcarriers for the Production of Tissue Plasminogen Activator (TPA 생산을 위한 섬유 세포의 미립 담체와의 접착성에 관한 연구)

  • 강재구;박형환;이현용
    • KSBB Journal
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    • v.4 no.2
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    • pp.104-109
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    • 1989
  • Among commercially available five different types of micro-carriers, collagen coated Cytodex III supports the best cell growth and production of Tissue type Plasminogen Activator from fibroblasts for all three various kinds of media(FBS, horse serum containing and serum free medium). 95% of attachment yield is obtained with Cytodex III in FBS containing medium compared to about 70% in serum free medium. However, higher production of TPA can be observed in horse serum containing and serum free media as $1.2\;\mu\textrm{g}/\;ml$ and $0.7 \mu\textrm{g}/\;ml$, respectively with $5.5{\times}10^6$ cells/ ml and $0.7{\times}10^6$ cells/ ml under perfusiom chemostat operations.

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Thrombolytic Therapy for Repeated Prosthetic Valvular Thrombosis That's Associated with Pregnancy under Heparin Anticoagulation (임신과 연관된 헤파린 항응고요법 중 반복적으로 발생한 인공판막 혈전증의 혈전용해치료)

  • Choe, Ju-Won;Kim, Yong-In L.
    • Journal of Chest Surgery
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    • v.41 no.4
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    • pp.484-488
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    • 2008
  • During long-term anticoagulation treatment with using heparin in a pregnant patient with a mechanical mitral prosthesis, we observed several anticoagulation-related complications, including repeated prosthetic valve thrombosis. This was found to be caused by heparin resistance due to an anti-thrombin III deficiency. Thrombolytic therapy using urokinase or tissue plasminogen activator (tPA) was successful and safe for her as well as her baby.

Storage of Pollen Biofactory for in vitro Growth and rProtein Synthesis (기내배양과 재조합 단백질 생산을 위한 화분 Biofactory의 저장기술의 개발)

  • 박희성;고재철
    • KSBB Journal
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    • v.19 no.3
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    • pp.174-177
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    • 2004
  • A method of collection and long-term storage of viable lily (Lilium longiflorum) pollen grains were developed for their in vitro growth and transformation in consistency. Petroleum ether, n-heptane, cyclohexane and benzene, as pollen collection medium, were determined less toxic to pollen growth in vitro than others tested. Pollen grains, however, lost their growth activity if stored in these solvents more than a week, So, a serial performance, that is, pollen grain collection in these solvents, air-drying and immediate transfer to low temperature condition was determined desirable for keeping the viability much longer. Pollen grains from this storage showed a successful transformation in vitro with a cDNA encoding tissue plasminogen activator (TPA) protein using Agrobacterium via vacuum infiltration according to western blotting analysis.

Effects of Acute Aerobic Exercise on Blood Pressure, Blood Lipids and Fibrinolytic Markers in Pre-hypertension College-aged Males (고혈압 전단계 대학생들의 단순 일회성 유산소 운동이 혈압, 혈중지질과 섬유소 용해 인자에 미치는 영향)

  • Kim, Daeyeol;Back, Kyoungyeop;Park, Hyeok;Lee, Hayan;Kim, Donghee
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.18 no.7
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    • pp.140-148
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    • 2017
  • The purpose of this study was to examine the effects of acute aerobic exercise at different intensities on the blood pressure, blood lipids and fibrinolytic markers in pre-hypertension college-aged males. Six subjects performed an acute running exercise at three different intensities(low intensity(LI): 50-60% heart rate reserve(HRR), moderate intensity(MI): 60-70%HRR, and high intensity(HI): 70-80%HRR). The blood pressure(systolic(SBP) and diastolic blood pressure(DBP), blood lipids(total cholesterol(TC) and high-density lipoprotein cholesterol(HDL-C)) and fibrinolytic markers(tissue plasminogen activator(tPA) and plasminogen activator inhibitor-1(PAI-1)) were determined before(PRE), immediately after(POST) and 60minutes after the exercise(60 POST). Results: the SBP in the LI group was significantly increased at POST(p=0.013). The ES levels for the SBP in the MI and HI groups were reduced (-1.33 and -1.23, respectively), though the differences were not significant. The HDL in the MI(p=0.003) and HI(p=0.002) groups were significantly increased at 60 POST. Also, the tPA in the MI(p=0.021) and HI(p=0.042) groups were significantly increased at POST.

Analysis of Plasminogen Activators Activity and Three Dimensional (3D) Culture of Endometrial Cells in Pigs (돼지 자궁내막 세포의 3차원 배양과 Plasminogen Activator 활성화 분석)

  • Cha, Hye-Jin;Lee, Sang-Hee;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Journal of Embryo Transfer
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    • v.28 no.3
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    • pp.273-280
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    • 2013
  • The aim of this study was to establish a three dimensional (3D) culture system of endometrial cells and to examine the plasminogen activators (PAs) activity in porcine uterine. The 3D culture system in porcine endometrial cells was composed to mixture 3D gel, stromal cells and epithelial cells. The 3D culture system was used to identify normal structure search as uterine tissue and PAs expression in this study. In results, porcine endometrium epithelial cells forming a top monolayer and endometrium stromal cells developed as fibroblast-like within 3D matrix scaffold. Expression of urokinase-type PA (uPA) and tissue-type PA (tPA) were observed during the 3D culture using immunofluorescence. PA activity in 3D-cultured endometrial cells was no significant difference between the tissue type, but 2D culture system were significantly lower than in 3D-cultured endometrial cells (P<0.05). Therefore, basic system and functional aspect of 3D culture could be established with similar system of endometrium tissue. We suggest that this study was assumed applicable as baseline data to investigate mechanism between porcine uterus cells in vitro.

Plasminogen Activator Inhibitor Type 1 Gene Polymorphism in Patients with Minimal Change Nephrotic Syndrome (소아 신증후군 환자에서 Plasminogen Activator Inhibitor Type 1 유전자 다형성)

  • Kim Young-Min;Hong Hyun-Kee;Kim Sung-Do;Cho Byoung-Soo
    • Childhood Kidney Diseases
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    • v.8 no.1
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    • pp.26-32
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    • 2004
  • Purpose : Hypercoagulability is present in patients with nephrotic syndrome. Plasminogen activator inhibitor type 1(PAI-1) is a major inhibitor of plasminogen activators. PAI-1 inactivates both tissue plasminogen activator(tPA) and urokinase plasminogen activator(uPA) by rapid formation of inactive 1:1 stoichiometric complexes. Recently some studies showed that the enhanced PAI-1 expression may be involved in the intraglomerular fibrinogen/fibrinrelated antigen deposition seen in nephrotic syndrome. Methods : PAI-1 gene promoter -844(G/A) polymorphism was evaluated in 146 children with minimal change nephrotic syndrome(MCNS) and 230 control subjects. The patients with MCNS were subdivided into 85 infrequent-relapser(IR) group and 61 frequent relapser(FR) group. PCR of PAI-1 gene promoter region including -844(G/A) and RFLP using the restriction enzyme Xhol were performed for each DNA samples extracted from the groups. Results : The distribution of PAI-1 genotype in the control group was G/G 81(32.5%), A/A 42(16.9%), and G/A 126(50.6%). The distribution of PAI-1 genotypes in the IR group of MCNS was G/G 29(34.1%), A/A 15(17.7%), and G/A 41(48.2%). The distribution of PAI-1 genotype in the FR group of MCNS was G/G 17(27.9%), A/A 18(29.5%), and G/A 26(42.6%). There was a significantly increased frequency of A/A genotype(P=0.0251) in the FR group of MCNS. Conclusion : Our results indicate that the PAI-1 gene promoter A/A genotype may be associated with the FR in MCNS.

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A novel variant of t-PA resistant to plasminogen activator inhibitor-1; expression in CHO cells based on In Silico experiments

  • Davami, Fatemeh;Sardari, Soroush;Majidzadeh-A, Keivan;Hemayatkar, Mahdi;Barkhordari, Farzaneh;Enayati, Somayeh;Adeli, Ahmad;Mahboudi, Fereidoun
    • BMB Reports
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    • v.44 no.1
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    • pp.34-39
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    • 2011
  • Resistance to PAI-1 is a factor which confers clinical benefits in thrombolytic therapy. The only US FDA approved PAI-1 resistant drug is Tenecteplase$^{(R)}$. Deletion variants of t-PA have the advantage of fewer disulfide bonds in addition to higher plasma half lives. A new variant was developed by deletion of the first three domains in t-PA in addition to substitution of KHRR 128-131 amino acids with AAAA in truncated t-PA. The specific activity of this new variant, $570\;IU/{\mu}g$, was found to be similar to those found in full length t-PA (Alteplase$^{(R)}$), $580\;IU/{\mu}g$. A 65% and 85% residual activity after inhibition by rPAI-1 was observed for full length and truncated-mutant form, respectively. This new variant as the first PAI-1 resistant truncated t-PA may offer more advantages in clinical conditions in which high PAI-1 levels makes the thrombolytic system prone to re-occlusion.

The Effect of Tissue Plasminogen Activator on TGF-${\beta}1$ Pre-Treated Human Mesothelial Cell Line (TGF-${\beta}1$으로 자극한 사람중피세포주에서 조직플라스미노겐 활성제가 미치는 영향)

  • Lee, Jung-Lim;Jeon, Soo-Jin;Yoo, Young-Choon;Kim, Ji-Hye;Lee, Yu-Mi;Kwon, Sun-Jung;Son, Ji-Woong;Choi, Eu-Gene;Na, Moon-Jun
    • Tuberculosis and Respiratory Diseases
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    • v.70 no.5
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    • pp.405-415
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    • 2011
  • Background: In an effort to find alternative therapeutic agents to prevent excessive fibrosis as a sequela to complicated parapneumonic effusion or empyema, we examined the effect of tissue plasminogen activator (tPA) as a fibrinolytic agent combined with talc or transforming growth factor (TGF)-${\beta}1$ in a human pleural mesothelial cell line, MeT-5A. Methods: MeT-5A cells were stimulated with various doses of talc, doxycycline or TGF-${\beta}1$ for 24 h and then were treated with tPA for an additional 24 h. Cell viability was measured by MTT assay. The production of interleukin (IL)-8 and vascular endothelial growth factor (VEGF) in the culture supernatants was measured by ELISA. Real-time PCR was carried out for measurement of type I collagen mRNA. Results: MeT-5A cells treated with talc showed a dose-dependent increase in production of IL-8. Talc also increased production of type I collagen mRNA at low doses, but talc did not influence the induction of VEGF. Addition of tPA to talc-stimulated cells showed further increases in the production of IL-8, but tPA did not influence the production of VEGF or type I collagen mRNA. TGF-${\beta}1$ increased the production of both VEGF and collagen type I mRNA, both of which were effectively inhibited by additional tPA treatment in MeT-5A cells. Conclusion: TGF-${\beta}1$ is a potent inducer of collagen synthesis without induction of IL-8 in MeT-5A cells. Addition of tPA after TGF-${\beta}1$ stimulation inhibited further fibrosis by direct inhibition of collagen mRNA synthesis as well as by inhibition of VEGF production.

Anticoagulant activities of piperlonguminine in vitro and in vivo

  • Lee, Wonhwa;Yoo, Hayoung;Ku, Sae-Kwang;Kim, Jeong Ah;Bae, Jong-Sup
    • BMB Reports
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    • v.46 no.10
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    • pp.484-489
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    • 2013
  • Piperlonguminine (PL), an important component of Piper longum fruits, is known to exhibit anti-hyperlipidemic, antiplatelet and anti-melanogenic activities. Here, the anticoagulant activities of PL were examined by monitoring activated-partial-thromboplastin-time (aPTT), prothrombin-time (PT), and the activities of thrombin and activated factor X (FXa). The effects of PL on the expressions of plasminogen activator inhibitor type 1 (PAI-1) and tissue-type plasminogen activator (t-PA) were also tested in tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) activated HUVECs. The results showed that PL prolonged aPTT and PT significantly and inhibited the activities of thrombin and FXa. PL inhibited the generation of thrombin and FXa in HUVECs. In accordance with these anticoagulant activities, PL prolonged in vivo bleeding time and inhibited TNF-${\alpha}$ induced PAI-1 production. Furthermore, PAI-1/t-PA ratio was significantly decreased by PL. Collectively, our results suggest that PL possesses antithrombotic activities and that the current study could provide bases for the development of new anticoagulant agents.

Effect of HRE and Bcl-2 on the Production of Plasminogen Activator in CHO cells

  • Bae, Geun-Won;No, Jeong-Gwon;Lee, Gyu-Min;Kim, Ik-Yeong;Kim, Ik-Hwan
    • 한국생물공학회:학술대회논문집
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    • 2002.04a
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    • pp.261-264
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    • 2002
  • CHO (Chinese hamster ovary) cells were transfected with plasmids containing both cis-acting HRE (hypoxia response element) and CMV-promoter that controls tissue-type plasminogen activator (t-PA). CHO cells with HRE produced 16.2 fold higher t-PA concentration than CHO cells without HRE. It was noted that hypoxia strongly induced CHO cell apoptosis. which resulted in decrease of cell viability and protein production. In this study. by introducing Bcl-2, anti-apoptotic gene, we tried to recover cell viability and increase the protein production. When batch culture of both control cells without transfection of Bcl-2 and cells transfected with Bcl-2 were performed in the absence of CoCl ι hypoxia mimic condition. the cells with Bcl-2 were effected specific cell growth rates, maximum cell density. Immunoblotting assay showed Bcl-2 was recombinant with HRE dependent t- P A expression cassette, and their expression level was depended on hypoxia. By introducing Bcl-2, both cell viability and maximum cell density could be increased.

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