• Title/Summary/Keyword: Thrombin

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The Ligand Occupancy of Endothelial Protein C Receptor Switches the Signaling Specificity of Thrombin from a Disruptive to a Protective Response in Endothelial Cells

  • Bae, Jong-Sup;Kim, Yong-Ung;Park, Moon-Ki
    • Proceedings of the Korean Environmental Sciences Society Conference
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    • 2008.11a
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    • pp.538-544
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    • 2008
  • Activated protein C (APC) is thought to exert antiinflammatory activities through the endothelial protein C receptor (EPCR)-dependent cleavage of protease activated receptor 1 (PAR-1) in endothelial cells. Since thrombin cleaves PAR-1 with $\sim$3-4-orders of magnitude higher efficiency, and PAR-1 is a target for proinflammatory activities of thrombin, it is not understood how APC can elicit protective responses through the cleavage of PAR-1. In this study, we demonstrate that EPCR is associated with caveolin-1 in endothelial lipid rafts, but its occupancy by protein C leads to its dissociation from caveolin-1 and subsequent recruitment of PAR-1 to protective signaling pathways through the coupling of PAR-1 to Gi-protein. When EPCR is bound by protein C, the PAR-1-dependent protective response in endothelial cells can be mediated by either thrombin or APC. These results provide a new paradigm for understanding the mechanism through which PAR-1 and EPCR participate in cellular signaling events in endothelial cells.

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Level of Platelet Derived Growth Factor(PDGF) in Blood Bank Platelet Concentrate (혈액은행 혈소판농축액의 혈소판유래성장인자 분비능)

  • Hong, Yong Taek;Han, Seung Kyu;Lee, Byung Il;Kim, Woo Kyung
    • Archives of Plastic Surgery
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    • v.33 no.6
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    • pp.732-736
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    • 2006
  • Purpose: The purpose of this pilot study was to investigate a potential of platelet concentrate obtained from blood bank(PCBB) in accelerating wound healing and to determine an effective treatment protocol by quantifying levels of platelet derived growth factor (PDGF)-BB in PCBB in vitro. Methods: The first study was designed to investigate quantity of PDGF-BB over stored time of the PCBB. The stored times for each PCBB were 1, 3, 5, 7, 9, 11 and 13 days. The second study was designed to determine efficacy of adding thrombin to stimulate release of PDGF-BB from the platelets of PCBB. The platelets were suspended and incubated in either with or without thrombin. On 30 minutes and days 1, 3, 5, 7 after incubation, the levels of PDGF-BB were measured. Results: PDGF-BB level showed a linear decrease over stored time of PCBB from the first day to the 13th day. Addition of thrombin increased PDGF-BB release from 30 minute through the 5th day. Conclusion: The results indicate that PCBB can provide sufficient amount of growth factors to stimulate wound healing and adding thrombin accelerate it.

Anticoagulant 1,2,3,4,6-Pentagalloyl-$\beta$-D-Glucopyranose Isolated from Geranium (Pelargonium inquinans Ait)

  • Ji Myeong-Sim;Piao Xiang-Lan;Jin Yu-Lan;Park Ro Dong
    • Archives of Pharmacal Research
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    • v.28 no.9
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    • pp.1037-1041
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    • 2005
  • Geranium (Pelargonium inquinans Ait) leaves were extracted with $80\%$ MeOH, and partitioned into n-hexane, ethyl acetate, BuOH and $H_2O$ to isolate the anticoagulant principles. The EtOAc fraction was found to be the most active, and was further purified using silica and octadecylisilane column chromatography employing a bioassay-guided fractionation method. The active compound was isolated and identified as $1,2,3,4,6-pentagalloyl-\beta-D-glucopyranose$(PGG) (compound I). The isolated anticoagulant significantly prolonged the activated partial thrombin time (APTT) and thrombin time (TT) using normal human plasma. One microgram of $1,2,3,4,6-pentagalloyl-\beta-D-glucopyranose$ showed 0.063 heparin units in the APTT and 2.73 heparin units in the TT for anti-thrombosis. This is the first report of the isolation of PGG from geranium plants.

Porcine Blood Plasma Transgluataminase Combined with Thrombin and Fibrinogen as a Binder in Restructured Meat

  • Tseng, Tsai-Fuh;Tsai, Chong-Ming;Yang, Jeng-Huh;Chen, Ming-Tsao
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.7
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    • pp.1054-1058
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    • 2006
  • The purpose of this study was to use pig blood plasma transglutaminase (TGase) combined with thrombin and fibrinogen as a binder, which was applied to restructured meat, and to investigate its effect on the restructured meat quality. Pig meat was obtained 10 h post mortem from a traditional market was ground using a 10 mm aperture plate. A binder admixture was added (TGase:thrombin:fibrinogen mixed as 0.5:1:20 (v/v/v) to which was added 12% of its volume of 0.25 M calcium chloride) at 0, 5, 10, 15 and 20% of meat weight. Measurements included cooking loss, shrinkage rate, shear value, total plate count, pH value, TBA value, color difference, tension strength and sensory evaluation. The results showed that ground meat containing 20% w/w of binder admixture had higher cooking loss, shrinkage rate and shear value (p<0.05). Addition of different percentages of binder admixture did not affect total plate count, pH value, TBA value, and sensory evaluation of restructured meat (p>0.05). Tension strength was increased with increased level of binder admixture. Addition up to 15% binder admixture to restructured meat showed better scores of sensory texture, flavor and total acceptability (p<0.05).

Effects of Lipophilic Fraction from Korean Red Ginseng on Platelet Aggregation and Blood Coagulation in Rats Fed with Corn Oil and Beef-tallow Diet (옥수수기름 및 우지 식이에 의한 혈소판 응집 반응과 혈액응고에 있어서 홍삼 지용성성분이 미치는 영향)

  • 이정희;박화진
    • Journal of Ginseng Research
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    • v.19 no.3
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    • pp.206-211
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    • 1995
  • This study was investigated to find the effects of petroleum ether extract (Lipophilic fraction) from Korean red ginseng on platelet aggregation and thrombin time of the plasma in two groups of the experimental rats. One group of rats were fed with 15% corn oil (15%kg-diet) containing a number of 18 : 2 (linoleic acid) or 15% beef-tallow (15%/kg-diet) containing saturated fatty acids for 3 weeks, and were followed by feeding the petroleum ether extract (25 mg/kg-diet) for 3 weeks. The other group of rats (control group) were fed with 15% corn oil or 15% beef-tallow for 6 weeks. The platelet aggregation induced by thrombin and collagen was significantly inhibited and the thrombin time was prolonged in the 15% corn oil plus petroleum ether extract administrated group than in the 15% corn oil administrated group. And the same results were shown in the 15% beef-tallow plus petroleum ether extract administrated group. These results suggest that the petroleum ether extract from Korean red ginseng may have the beneficial effects on the inhibition of the platelet aggregation and the inhibition of blood coagulation induced by dietary fats.

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Activation of Platelet Rich Plasma by Soluble Canine Small Intestinal Submucosa Gel and Bovine Thrombin (개 소장점막하 겔과 소 트롬빈을 이용한 혈소판풍부혈장의 활성화 연구)

  • Lee, A-Jin;Lee, Changsun;Kim, Hyun;Chung, Dai-Jung;Do, Sun Hee;Kim, Hwi-Yool
    • Journal of Veterinary Clinics
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    • v.31 no.3
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    • pp.170-174
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    • 2014
  • This study evaluated the efficacy of soluble canine small intestinal submucosa gel in comparison to bovine thrombin in activating rabbit platelet rich plasma (PRP) by detecting growth factors. PRP from rabbits was activated by using soluble canine SIS gel, bovine thrombin, or both. The surface morphology of each group of samples was examined by scanning electron microscopy. The release of transforming growth factor (TGF)-${\beta}1$ from each set of samples was measured over 7 days using enzyme-linked immunosorbent assay. The PRP-canine SIS gel group exhibited the highest total amount of released TGF-${\beta}1$. However, there were no significant differences between any groups. The use of soluble type of canine SIS gel could be an effective alternative to bovine thrombin.

Percutaneous Thrombin Injection Based on Computational Fluid Dynamics of Femoral Artery Pseudoaneurysms

  • Hyoung-Ho Kim;Kyung-Wuk Kim;Changje Lee;Young Ho Choi;Min Uk Kim;Yasutaka Baba
    • Korean Journal of Radiology
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    • v.22 no.11
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    • pp.1834-1840
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    • 2021
  • Objective: To analyze the computational fluid dynamics (CFD) of femoral artery pseudoaneurysm (FAP), identify a suitable location and timing for percutaneous thrombin injection (PTI) based on this analysis, and report our clinical experience with the procedure. Materials and Methods: CFD can be used to analyze the hemodynamics of the human body. An analysis using CFD recommended that the suitable location of the needle tip for PTI is at the center of the aneurysm sac and the optimal timing for starting PTI is during the early inflow phase of blood into the sac. Since 2011, seven patients (three male and four female; median age, 60 years [range, 43-75 years]) with FAP were treated with PTI based on the devised suitable location and time. Prior to the procedure, color Doppler ultrasonography was performed to determine the location and timing of the thrombin injection. Results: The technical success rate of the PTI was 100%. The amount of thrombin used for the procedure ranged from 200 IU to 1000 IU (median, 500 IU). None of the patients experienced any symptoms or signs of embolic complications during the procedure. Follow-up CT images did not reveal any embolism in the lower extremities and showed complete thrombosis of the pseudoaneurysm. Conclusion: Based on our study of CFD, PTI administered centrally in the FAP during early inflow, as seen on color Doppler, can be an effective technique.

Aptamer-Based Precipitation as an Alternative to the Conventional Immunoprecipitation for Purification of Target Proteins

  • Song, Seongeun;Cho, Yea Seul;Lee, Sung-Jae;Hah, Sang Soo
    • Bulletin of the Korean Chemical Society
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    • v.35 no.9
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    • pp.2665-2668
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    • 2014
  • Aptamers are oligonucleotides or peptide molecules that are able to bind to their specific target molecules with high affinity via molecular recognition. In this study, we present development of aptamer-based precipitation assays (or simply aptamoprecipitation) for His-tagged proteins and thrombin to compare their purification efficiency with other conventional affinity precipitation methods. A crosslinking method was employed to immobilize thiol-functionalized aptamers onto the surface of polystyrene resins, enabling them to specifically bind to His-tag and to thrombin, respectively. The resulting aptamer-functionalized resins were successfully applied via a one-step experiment to purification of His-tagged proteins from complex E. coli and to thrombin extraction, exhibiting superior or at least comparable purification results to the conventional immobilized metal affinity precipitation or immunoprecipitation.

Effects of Benzoquinone on Aggregation and Cytotoxicity in Platelets (Benzoquinone에 의한 혈소판 응집 억제 및 세포독성)

  • 이선구;강규태;이무열;정승민;정진호
    • Toxicological Research
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    • v.16 no.4
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    • pp.311-315
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    • 2000
  • Previous studies showed that benzoquinone derivatives inhibited platelet aggregation. but there is no information available on their cytotoxicity to platelets. 1n the present study. washed platelets isolated from rats were treated with 1.4-benzoquinone. a representative benzoquinone derivative. to examine its antiaggregating effect and cytotoxicity. 1.4-Benzoquinone significantly inhibited thrombin-induced platelet aggregation. Consistent with this finding. 1.4-benzoquinone suppressed cytosolic calcium increase induced by thrombin. To examine the cytotoxicity by 1 A-benzoquinone in platelets. turbidometry and lactate dehydrogenase release were measured. Treatment with 1.4-benzoquinone resulted in slight cytotoxicity (30% release at 60 min) to platelets. However. the cytotoxicity was not correlated with increase of cytosolic calcium levels in platelets. All these data suggested that 1.4-benzoquinone inhibited thrombin-induced platelet aggregation mediated by inhibition elf calcium level increase and that 1.4-benzoquinone reveals cytotoxicity to some extent without alteration of calcium level in platelets.

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Anticoagulant activities of curcumin and its derivative

  • Kim, Dong-Chan;Ku, Sae-Kwang;Bae, Jong-Sup
    • BMB Reports
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    • v.45 no.4
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    • pp.221-226
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    • 2012
  • Curcumin, a polyphenol responsible for the yellow color of the curry spice turmeric, possesses antiinflammatory, antiproliferative and antiangiogenic activities. However, anticoagulant activities of curcumin have not been studied. Here, the anticoagulant properties of curcumin and its derivative (bisdemethoxycurcumin, BDMC) were determined by monitoring activated partial thromboplastin time (aPTT), prothrombin time (PT) as well as cell-based thrombin and activated factor X (FXa) generation activities. Data showed that curcumin and BDMC prolonged aPTT and PT significantly and inhibited thrombin and FXa activities. They inhibited the generation of thrombin or FXa. In accordance with these anticoagulant activities, curcumin and BDMC showed anticoagulant effect in vivo. Surprisingly, these anticoagulant effects of curcumin were better than those of BDMC indicating that methoxy group in curcumin positively regulated anticoagulant function of curcumin. Therefore, these results suggest that curcumin and BDMC possess antithrombotic activities and daily consumption of the curry spice turmeric might help maintain anticoagulant status.