• Title/Summary/Keyword: heparin

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Comparison of Two Methods for Heparin Sensitivity; Activated Partial Thromboplastin Time Assay using in vitro Heparin-spiked Sample and Anti-Xa Assay using in vivo Heparin-treated Sample

  • Koo, Bon-Kyung;Kwon, Eui-Hoon;Ryu, Kwang-Hyun;Yun, Jae-Won;Kim, Hee-Jin
    • Korean Journal of Clinical Laboratory Science
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    • v.43 no.4
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    • pp.133-137
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    • 2011
  • The monitoring of heparin therapy is using almost aPTT assay. This study is compare to estimating aPTT therapeutic range using in vitro heparin-spiked sample and aPTT therapeutic range using in vivo heparin-treated sample. Normal pooled plasma was collected from 20 healthy representative individuals. 11 concentration of heparinized plasmas from 0 U/mL to 1.0 U/mL at intervals of 0.1 U/mL made by addition of heparin to normal pooled plasma were measured aPTT. The aPTT therapeutic range was performed through correlation analysis between heparin level 0.2 to 0.4 U/mL and aPTT. 30 plasmas from patients on heparin therapy were measured aPTT and anti-Xa activity. The aPTT therapeutic range was performed through correlation analysis between anti-Xa activity 0.3 to 0.7 U/mL and aPTT. The aPTT therapeutic range corresponded by heparin level-vs-aPTT value regression analysis was 60.7 to 102.4 seconds. The aPTT therapeutic range corresponded by anti-Xa activity-vs-aPTT value regression analysis was 85.3 to 147.5 seconds. The validation of heparin sensitivity using in-vitro heparin sample was not considered. The establishing aPTT therapeutic range is recommended anti-Xa activity using in-vivo sample.

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Purification and Characterization of Heparin Lyase I from Bacteroides stercoris HJ-15

  • Kim, Wan-Seok;Kim, Byung-Taek;Kim, Dong-Hyun;Kim, Yeong-Shik
    • BMB Reports
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    • v.37 no.6
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    • pp.684-690
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    • 2004
  • Heparin lyase I was purified to homogeneity from Bacteroides stercoris HJ-15 isolated from human intestine, by a combination of DEAE-Sepharose, gel-filtration, hydroxyapatite, and CM-Sephadex C-50 column chromatography. This enzyme preferred heparin to heparan sulfate, but was inactive at cleaving acharan sulfate. The apparent molecular mass of heparin lyase I was estimated as 48,000 daltons by SDS-PAGE and its isoelectric point was determined as 9.0 by IEF. The purified enzyme required 500 mM NaCl in the reaction mixture for maximal activity and the optimal activity was obtained at pH 7.0 and $50^{\circ}C$. It was rather stable within the range of 25 to $50^{\circ}C$ but lost activity rapidly above $50^{\circ}C$. The enzyme was activated by $Co^{2+}$ or EDTA and stabilized by dithiothreitol. The kinetic constants, $K_m$ and $V_{max}$ for heparin were $1.3{\times}10^{-5}\;M$ and $8.8\;{\mu}mol/min{\cdot}mg$. The purified heparin lyase I was an eliminase that acted best on porcine intestinal heparin, and to a lesser extent on porcine intestinal mucosa heparan sulfate. It was inactive in the cleavage of N-desulfated heparin and acharan sulfate. In conclusion, heparin lyase I from Bacteroides stercoris was specific to heparin rather than heparan sulfate and its biochemical properties showed a substrate specificity similar to that of Flavobacterial heparin lyase I.

Effects of Salt and Heparin on the Activation of Lipoprotein Lipase(LPL)

  • Kim, Tae-Woong;Lee, Jae-Bok;Son, Heung-Soo;Kim, Sung-Wan
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.24 no.3
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    • pp.355-361
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    • 1995
  • Lipoprotein lipase(LPL) is an acylglycerol hydrolase and is the extrahepatic enzyme responsible for the hydrolysis of triglyceride-rich plasma lipoproteins. LPL has been isolated from bovine milk by affinity chromatography on heparin-sepharose in 2M NaCl, 5mM barbital buffer, pH 7.4. Para-nitrophenyl butyrate(PNPB) was used as a substrate for the determination of LPL activity. Molecular weight of LPL was 55KD on 10% SDS-PAGE. When the effects of heparin on LPL activation were compared, LPL activity of heparin added group increased approximately 5 times higher than that of heparin non-added groups. These results indicated that heparin involved in the stabilization of LPL structure that led to increase enzyme activity. Furthermore, LPL activity increased about 4 times compared to the absence of heparin at various pH. LPL was stabilized when heparin was added either low or high salt concentrations. With the presence of heparin, NaCl concentration did not affect LPL activity at pH range 6∼9.

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Heparin-induced Thrombocytopenia Type II after Free Flap Operation

  • Baek, Jiwoong;Park, Jung Hyun;Cha, In-Ho;Kim, Hyung Jun
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.35 no.6
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    • pp.408-411
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    • 2013
  • After radical excision of a tumor in the maxillofacial area, functional and esthetic reconstruction is needed, including flap surgery. Among the many etiologies of flap failure, venous thrombosis is one of the most frequent. Heparin is used routinely in the effort to avoid development of venous thrombosis. In rare cases, heparin-induced thrombocytopenia (HIT) type II occurs due to exposure to heparin. Heparin attached to platelet factor 4 forms a PF4/heparin-immunoglobulin G immune complex on platelet surfaces. This complex activates platelets, which leads to multiple coagulation in venous and arterial blood. We report here on a rare occurrence of HIT type II following fibula free flap surgery.

Does Low-Dose Heparin Have a Significant Role in Free Flap Surgery?

  • An, Mun-Young;Shin, Jin Yong;Lee, Young-Keun;Sabbagh, M. Diya;Roh, Si-Gyun;Lee, Nae-Ho
    • Archives of Craniofacial Surgery
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    • v.18 no.3
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    • pp.162-165
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    • 2017
  • Background: It is controversial issue that heparin decreases thrombosis for microsurgical anastomosis, and its effective role is under discussion. This study is for proving whether low-dose heparin is preventing thrombosis in free flap reconstruction. Methods: Through chart reviews of 134 patients, using low-dose heparin for free tissue transfer from 2011 to 2016, retrospective analysis was performed. 33 patients received low-dose heparin therapy after surgery. And 101 patients received no-heparin therapy. Complications included flap necrosis, hematoma formation, dehiscence and infection. Results: In no-heparin therapy group, comparing the flap necrosis revealed 16 cases (15.84%). And, flap necrosis was 6 cases (18.18%) in low-dose heparin therapy group. The statistical analysis of flap necrosis rate showed no significant difference (p=0.75). The results showed that there was no significant difference of flap necrosis rate between two groups. Conclusion: In this study, patients in the low-dose heparin group had no significantly lower rates of flap failure compared with no-heparin group. This suggests that low-dose heparin may not prevent thrombosis and subsequent flap failure to a significant extent.

Modified Cellulose for Heparin Binder (헤파린 제거제용 셀룰로오스의 개질)

  • 이원규;박기동
    • Journal of Biomedical Engineering Research
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    • v.15 no.3
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    • pp.259-264
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    • 1994
  • Heparin binders (Cell-PALA) used for selective heparin removal from blood, were prepared by immobilizing a cationic polymer, poly(allylamine) (PALA), onto cellulose substrate by a novel method. Their absorbing capacity for heparin was compared with untreated cellulose control using heparin solution in vitro. The surface areas of obtained heparin binders and untreated cellulose were 1.36 and ($2.56{\mu} g$/$cm^2$, respectively. The amount of bound heparin to PALA immobilized celluloses was determined to be 0.16 - $0.30{\mu}g$/cm, which is much higher than that of untreated cellulose ($0.03{\mu} g$/$cm^2$). These results suggest that Cell-PALA materials can be utilized for a heparin removal system.

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Studies on the Effects of the Capacitation Methods of Spermatozoas on in-vitro Fertilization and Cleavage Rate of Bovine Follicular Oocytes (수정능획득 처리법이 소 난포란의 체외수정 및 분할율에 미치는 영향에 관한 연구)

  • 김상근;한성욱;한방근
    • Korean Journal of Animal Reproduction
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    • v.15 no.2
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    • pp.125-132
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    • 1991
  • The studies on the carried out to investigate the effects of capacitation method of spermatozoa on the in vitro fertilization and cleavage rate of bovine follicular oocytes. The ovaries were obtained from slaughtered Korean native cows. The follicular oocytes surrounded with cumulus cells were recovered by aspirating follicular fluids from the visible of diameter 3~5mm. The follicular oocytes were cultured in TCM-199 medium containing hormones and FCS for 24~48hrs in an incubator with 5% CO2 in air at 38.5$^{\circ}C$ and then matured oocytes were again cultured for 12~18hrs with motile capacitated sperm by preincubation of mKRB, treatment of HIS(high strength ion), Ca-IA(Inophore A), BFF(bovine follicular fluids) and heparin. The results obtained in these experiments were summarized as follows : 1. The in vitro fertilizatin and cleavage rate offollicular oocytes fertilized with capacitated spermatozoas in BO solution by preincubation of mKRB, treatment of HIS, Ca-IA, BFF and heparin method were 53.1%, 33.9%, 50.8%, 48.1%, 58.8% and 28.1%, 17.7%, 26.2%, 22.8%, 32.8%, respectively. And the fertilization and cleavage rate of heparin method was of highest of all. 2. The in vitro fertilization and cleavage rate of follicular oocytes fertilized with capacitated spermatozoas in BO solutin by both caffeine, BSA and heparin methods were 65.8%, 70.3% and 40.8%, 47.3%, respectively, and those rates were higher treatment of heparin+BSA, heparin+caffeine than treatment of heparin. 3. The in vitro fertilization and cleavage rate of follicular oocytes fertilized with capacitated spermatozoa in BO solution with heparin concentrations of 2, 5, 10, 20, 40$\mu\textrm{g}$/ml were 50.0%, 54.7%, 58.1%, 51.7% and 27.9%, 32.8%, 37.1%, 30.0%, respectively. And the fertilization and cleavage rate of follicular oocytes fertilized with capacitated spermatozoas in BO solution with 10$\mu\textrm{g}$/ml of heparin was the highest of all. 4. The in vitro fertilization and cleavage rate of follicular oocytes fertilized with capacitated spermatozoas in BO solution containing heparin with caffeine concentraton of 10, 20, 30, 40$\mu\textrm{g}$/ml were 71.4%, 74.3% and 70.6%, 70.0% and 45.7%, 47.3%, 44.1%, 41.4%, respectively. The fertilization and cleavage rate of spermatozoa fertilized in BO solution with caffeine and heparin together(70.3~74.3%) was higher than that of spermatozoa fertilized in BO solution with heparin(58.8%). 5. The in vitro fertilization and cleavage rate of follicular oocytes fertilized with capacitated spermatozoas in BO solution containing heparin with BSA concentration of 5, 10, 20, 30$\mu\textrm{g}$/ml were 63.6%, 62.9%, 66.7%, 60.3% and 44.1%, 43.5%, 48.5%, 42.7%, respectively. The fertilization and cleavage rate of spermatozoa fertilized in BO solution with BSA and heparin together(60.3~66.73%) was higher than that of spermatozoa fertilized in BO solution with heparin(58.8%).

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Control of heparinization by activated clotting time during extracorporeal circulation (개심술시 Activated Clotting Time 을 이용한 Heparin 투여 조절에 관한 임상적 고찰)

  • 서충헌
    • Journal of Chest Surgery
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    • v.16 no.3
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    • pp.281-288
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    • 1983
  • Heparinization is an essential step in extracorporeal circulation for open heart surgery. But wide individual variation to heparin effect sometimes makes it difficult to anticoagulate safely or neutralize appropriately. Because the conventional set protocol of heparinization did not consider this individual variation, a new method of control of heparinization was proposed by Dr. Brian Bull in 1974. We compared the group in which a conventional set protocol was used [Control group] with the other in which a new protocol modified from that of Bull was used [ACT group], on the aspects of the dosages of heparin and protamine administered and postoperative bleeding. Our conventional protocol [Control group] consisted of: 1. Initial heparin was given at dose of 350U/Kg into the right atrium prior to bypass. 2. Additional heparin was given every hour during E.C.C., as much as a half of the Initial dose. 3. 600U of heparin was mixed into every 100ml. of priming solution. 4. The protamine dose was calculated by totalling the units of heparin given to the patient and giving 1 .8mg. of protamine per 100 units of heparin. ACT protocol [ACT group] consisted of: 1. Initial heparinization was same as that of conventional protocol. 2. ACT`s were checked before [A point] and 10 minutes after initial heparinization [B point]. With these 2 points, a dose response curve was drawn. 3. Heparin for the priming solution was same as in control group. 4. Every 30 minutes during E.C.C., ACT`s were checked with Hemochron [International Technidyne Corp.]. ACT between 450 and 600 seconds was regarded as safety zone. If ACT checked at a time was below 450 seconds, heparin dose was calculated on the dose-response curve to lengthen ACT to 480 seconds and was given into the oxygenator. 5. About 10 minutes before the term of E.C.C., ACT was checked to estimate the blood heparin level at the time. Then, protamine dose was calculated at dose of 1.Stag per 100 units of heparin. The calculated dose of protamine was mixed into 50 to lO0ml of 5% Dextrose Water and dripped intravenously during the period of 15 minutes. Compared these two groups mentioned above, results were obtained as follows: 1. Mean value of normal ACT checked with Hemochron on 30 preoperative patients was 124 seconds [range 95-145 sec.]. 2. Doses of heparin and protamine given to the patient were decreased in ACT group as much as 32.2% and 62.2% respectively. 3. Postoperative bleeding and transfusion were also decreased in ACT group in 60.5% and 67.1% respectively. 4. Our modified dose-response curve did not cause any problems in the control of heparinization. 5. Initial heparinization [Heparin 350U/Kg] was sufficient for the most patients until 60 minutes under extracorporeal circulation. 6. We used 1.5mg of protamine to neutralize 100 units of heparin. But smaller dose of protamine may be sufficient for appropriate neutralization.

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Heparin Release from Hydrophobic Polymers : (I) In Vitro Studies

  • Kim, Sung-Ho;Kim, Sung-Wan
    • Archives of Pharmacal Research
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    • v.9 no.4
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    • pp.193-199
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    • 1986
  • The release of heparin from monolithic devices composed of different ratios of polythylene oxide (PEO MW 20,000) and hydrophobic polydimethylsiloxane or polyurethane was investigated. Water soluble PEO blended into the polymers provided a controlled release of heparin. The release rate of heparin could be controlled by varying the content of PEO. The heparin release rate from the devices increased as the content of PEO in the devices increased. The release mechanism may be associated with the creation of pore of domain through the devices following the swelling and the change in the physical structure of the polymer network. Hydrophobic polydimethylsiloxanes and polyurethanes containing PEO can provide an antith rombogenic material for prologed release of heparin from blended system.

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Heparin Release from Polyurethane-Polyethylene Oxide-Polypropylene Oxide Device (폴리우레탄-폴리에칠렌옥사이드-폴리프로필렌옥사이드-디바이스로부터의 헤파린 방출)

  • Yoo, Jae-Gwon;Jun, Sung-Joo;Kim, Sung-Ho
    • Journal of Pharmaceutical Investigation
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    • v.18 no.4
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    • pp.169-174
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    • 1988
  • The release of heparin from monolithic devices composed of different ratios of polyethylene oxide-polypropylene oxide (PEO-PPO) and hydrophobic polyurethane was investigated. The release rate of heparin could be controlled by varying the PEO-PPO content. The heparin release rate from the devices increased as the content of PEO-PPO in the devices increased. The release mechanism may be associated with creation of micro-channels and pores through the devices following the change in the physical structure of the polymer network. Hydrophobic polyurethane containing PEO-PPO can provide an antithrombogenic material for prolonged release of heparin from a heparin blended system.

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