• Title/Summary/Keyword: heparin

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Purification of Progelatinase A (Matrix Metalloproteinase 2) and a Tissue Inhibitor of Metalloproteinase-2(TIMP-2) from T98G Human Glioblastoma Cells

  • Lee, Ho-Jae;Chung, Myung-Chul;Lee, Choong-Hwan;Chun, Hyo-Kon;Kho, Yung-Hee
    • BMB Reports
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    • v.28 no.1
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    • pp.33-39
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    • 1995
  • The Gelatinases (typeIV collagenases) are metalloproteinases that may play an important role in tumor invasion and metastasis. Progelatinase A was purified from a conditioned medium of T98G human glioblastoma cells. TIMP-2 complexed progelatinase A and free progelatinase A were separated by heparin affinity HPLC. The final product was homogeneous on SDS-PAGE, with a molecular weight of 64,000 daltons without reduction. TIMP-2 and free progelatinase A were separated from TIMP-2 complexed progelatinase A by reverse-phase HPLC in the presence of trifluoroacetic acid. TIMP-2 complexed progelatinase A was resistant to activation by p-aminophenyl mercuric acetate (APMA), and showed less than 20% of the activity of the TIMP-2 free active enzyme. TIMP-2 free progelatinase A was easily activated to the mature form with a molecular weight of 57,000 daltons by APMA and showed high activity compared to the TIMP-2 complexed enzyme.

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Resistance Mechanism of Acinetobacter spp. Strains Resistant to DW-116, a New Quinolone

  • Choi, Keum-Hwa;Baek, Moon-Chang;Kim, Byong-Kak;Choi, Eung-Chil
    • Archives of Pharmacal Research
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    • v.21 no.3
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    • pp.310-314
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    • 1998
  • DW-116 is a new fluoroquinolone antimicrobial agent with a broad spectrum. In order to elucidate the resistance mechanism to DW-116 in Acinetobacter spp. bacteria, total chromosomal DNA was isolated from 10 strains of Acinetobacter spp. resistant to DW-116. Quinolone resistance determinant region (QRDR) of DNA gyrase gene was amplified by PCR. The 345 bp nucleotide fragment yielded was inserted into pKF 3 which was used as the vector. Comparisons of the DNA sequences of 8 strains with that of the wild type strain revealed a Ser-83 to Leu mutation in mutants and all ten strains contained one silent mutation$(T{\rightarrow}G)$in QRDR. From Acinetobacter MB4-8 strain, DNA gyrase was isolated and purified, through novobiocin-sepharose, heparin-sepharose affinity column chromatography. The enzyme was composed of two subunits and the molecular mass of subunits A and B were 75.6 and 51.9 kDa, respectively. The supercoiling activity of the reconstituted DNA gyrase composed of subunit A from Acinetobacter MB4-8 and subunit B from E. coli was not inhibited by $128{\mu}\textrm{g}$ml of ciprofloxacin. It might be said that one of the resistance mechanisms to DW-116 in Acinetohacter MB4-8 was subunit A alteration of DNA gyrase.

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Metabolomics Investigation of Cutaneous T Cell Lymphoma Based on UHPLC-QTOF/MS

  • Zhou, Qing-Yuan;Wang, Yue-Lin;Li, Xia;Shen, Xiao-Yan;Li, Ke-Jia;Zheng, Jie;Yu, Yun-Qiu
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.13
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    • pp.5417-5421
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    • 2014
  • Objectives: The identification of cutaneous T cell lymphoma (CTCL) biomarkers may serve as a predictor of disease progression and treatment response. The aim of this study was to map potential biomarkers in CTCL plasma. Design and Methods: Plasma metabolic perturbations between CTCL cases and healthy individuals were investigated using metabolomics and ultrahigh performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-QTOF/MS). Results: Principal component analysis (PCA) of the spectra showed clear metabolic changes between the two groups. Thirty six potential biomarkers associated with CTCL were found. Conclusions: Based on PCA, several biomarkers were determined and further identified by LC/MS/MS analysis. All of these could be potential early markers of CTCL. In addition, we established that heparin as a nticoagulant has better pre-treatment results than EDTA with the UHPLC-QTOF/MS appraoch.

Neuronal Differentiation of PC12 Cells Cultured on Growth Factor-Loaded Nanoparticles Coated on PLGA Microspheres

  • Park, Keun-Hong;Kim, Hye-Min;Na, Kun
    • Journal of Microbiology and Biotechnology
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    • v.19 no.11
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    • pp.1490-1495
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    • 2009
  • The development of nanotechnology has penetrated the fields of biology and medicine, resulting in remarkable applications for tissue regeneration. In order to apply this technology to tissue engineering, we have developed nano-scaled 3D scaffolds consisting of growth factor-loaded heparin/poly(l-lysine) nanoparticles (NPs) attached to the surface of polymeric micro spheres via polyionic complex methods. Growth factor-loaded NPs were simply produced as polyelectrolyte complexes with diameters of 100-200 nm. They were then coated onto positively charged poly(lactic-co-glycolic acid) (PLGA) pretreated with polyethyleneimine to enable cell adhesion, proliferation, and stimulation of neurite outgrowth. Propidium iodide staining and $\beta$-tubulin analysis revealed that neuronal PC12 cells proliferated extensively, expressed significant amounts of b-tubulin, and showed well-structured neurite outgrowth on polymeric microspheres by stimulation with growth factors. These results suggest that cellular adhesion and biological functionality on prepared PLGA microspheres enabled terminal differentiation of neuronal cells.

Effect of Bovine Oviductal Epithelial Cell(BOEC) Conditioned Medium on In Vitro Development of Bovine Embryos Fertilized In Vitro (난관상피세포 Conditioned Medium이 체외수정된 소 수정란의 체외 발달에 미치는 영향)

  • 오종훈;김동훈;정형민;이훈택;정길생
    • Korean Journal of Animal Reproduction
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    • v.17 no.1
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    • pp.69-74
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    • 1993
  • This study was investigated to examine the effect of conditioned medium from bovine oviductal cell(BOEC) in the co-culture system with BOEC on in vitro development of in vitro produced bovine embryos. Oocyte-cumulus complexes were cultured for 24 hrs in TCM-199 supplemented with 10% fetal calf serum, 1$\mu\textrm{g}$/ml FSH and 21U hCG, 1$\mu\textrm{g}$/ml oestradiol-17$\beta$ at 39$^{\circ}C$ under 5% CO2 in air. In vitro fertilization was performed with epididymal sperm and heparin (10$\mu\textrm{g}$/ml, 15min.) or caffeine(2.5mM)-treated spermatozoa. Oocytes were incubated with 1$\times$106 spermatozoa/ml for 18 hrs and then cultured in various culture system for 7 days. The development rates of 16-cell or blastocyst stages were recorded on 4, 7 days, respectively, after incubating. The proportions ofembryonic development into molulae and blastocysts were higher in cumulus cell co-culture(23.4%) and BOEC co-culture(34.3%) than in M199-FCS(6.1%). Similarily, the development rates into molulae and blastocysts were significantly higher in BOEC-conditioned medium than those in M199-FCS. Therefore, it is suggested that BOEC co-culture and BOEC conditioned medium increase significantly the development of in vitro produced bovine embryos in in vitro system.

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Production of Korean Native Calf by In Vitro Maturation, Fertilization, Cultivation and Transfer of Embryos into Holstein Cows (체외성숙, 수정 및 배양된 한우 체외수정란의 유우이식에 의한 산자 생산)

  • 박충생
    • Korean Journal of Animal Reproduction
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    • v.18 no.1
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    • pp.47-54
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    • 1994
  • The objective of this study was to produce Korean native calves following transfer of in vitro matured, fertilized and cultured embryos into Holstein cows. The ovaries of Korean native cows or heifers were obtained from an abattoir and kept on 25 to 28$^{\circ}C$ and transported to laboratory within 2 hrs. The oocytes were matured in vitro (IVM) for 24 hrs in TCM-199 supplemented with 35$\mu\textrm{g}$/ml FSH, 10$\mu\textrm{g}$/ml LH, 1$\mu\textrm{g}$/ml estradiol-17$\beta$ and granulosa cells at 39$^{\circ}C$ under 5% CO2 in air. They were fertilized in vitro (IVF) by epididymal spermatozoa treated with heparin for 24 hrs., and then the zygotes were co-cultured in vitro (IVC) with bovine oviductal epithelial cells for 7 to 9 days. Late morulae and blastocysts produced in vitro were nonsurgically transferred to recipient cows by unilaterial. Recipients were monitored for estrus and for pegnancy by rectal plapation in 60 days after embryo transfer. One of them was pregnant to term and produced a female weighing 42.5kg at birth.

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Evaluation of Blood compatibility at lumbrokinase immobilized polymer valves in vivo (룸브로키나제가 고정화된 폴리머 밸브의 invivo 혈액적합성 평가)

  • Park, Yong-Doo;Ryu, Eun-Sook;Kim, Jong-Won;Min, Byoung-Goo
    • Proceedings of the KOSOMBE Conference
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    • v.1998 no.11
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    • pp.227-228
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    • 1998
  • Lumbrokinase, potent fibrinolytic enzyme purified from earthworm, was immobilized onto polyurethane valves using photoreaction, photoreactive polyallyl-amino as a photoreactive linker. For evaluation of blood compatibility, lumbrokinase immobilized polymer valves were assembled into the total artificial heart (TAH). This TAH was implanted to 60kg healthy lamb for 1-3 days with the cardiac output 5 L/min. In the control lamb, the valves were untreated, in ore other, only valves on the right were treated, and in the remaining animal, only those on the left. To facilitate the thrombus formation, low doses of heparin were administered. For evaluation of the immobilized lumbrokinase, thrombus formation, proteolytic and fibrinolytic activity was measured. This data shows that lumbrokinase-treated polyurethane valves lead to decreased thrombus formation in vivo, and that their biocompatibility is therefore higher than that of untreated valves.

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Clinical experience of open heart surgery -500 cases- (개심술 500예에 대한 임상적 고찰)

  • 정황규
    • Journal of Chest Surgery
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    • v.19 no.4
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    • pp.633-643
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    • 1986
  • Five hundred cases of open heart surgery were operated in the Dept. of Thoracic and Cardiovascular Surgery, Pusan National University Hospital from July, 1981 to October, 1986. The clinical data were summarized as follows: 1. The age distribution of congenital heart disease patients was 2 to 41 years old and mean age was 13.4 years and of acquired heart disease was 11 to 57 years old and mean age was 32.7 years. 2. There were 319 cases [63.8%] of acyanotic congenital heart anomalies, 56 cases [11.2%] of cyanotic anomalies and 125 cases [25.[%] of acquired heart disease. 3. For myocardial protection, Bretschneider and potassium glucose solution had been used as cardioplegic solution and since 1983, GIK solution is being used repeatedly every 30 to 40 minutes time interval with excellent results. 4. The ingredient of the priming solution is Hartmann`s solution, mannitol, sodium bicarbonate, potassium, chloride, fresh ACD whole blood, calcium chloride, heparin and dexamethasone. 5. There were 94 cases of mild hypothermia, 280 cases of moderate hypothermia and 126 cases of intermediate hypothermia. 6. The overall mortality was 8.2%. And the mortality rate in each disease entity is 2.5% in acyanotic congenital cases, 33.9% in cyanotic congenital cases and 11.2% in acquired heart disease.

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The Effect of Temperature of Cardioplegic Soultion on Myocardial Protection from Ischemia - Experimental Study using Isolated Rat Heart Perfusion Technique - (흰쥐의 적출된 심장에서 심정지액의 온도가 심근보호에 미치는 영향)

  • 김용한
    • Journal of Chest Surgery
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    • v.25 no.2
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    • pp.131-136
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    • 1992
  • The effect of temperature of cardioplegic solution on myocardial preservation was studied using isolated rat heart perfusion technique. Twenty Sprague-Dawley rats, weighing 120~140gm, were pretreated with intraperitoneal injection of heparin sodium[300u/kg] and then the hearts were excised after cervical herniation 30 minutes later. The hearts were perfused in isolated working heart apparatus with oxygenated modified Tyrode solution at 37oC. After 10 minutes of non working heart perfusion, the hearts were subjected to arrest for 30 minutes by administration of 5cc cardioplegic solution at the temperature of 4oC [Group I ], 15oC [Group II], 25oC [Group III], 37oC[Group IV]. At the same time, the topical cooling of heart was performed using ice saline. After arrest, the hearts were reperfused by non working heart perfusion for 1 hour with modified Tyrode solution at 37oC. The CPK, GOT and LDH in reperfusate were measured at 5,20,40,60 minutes after start of reperfusion. With the values of those, we compared the effect of temperature of cardioplegic solution on myocardial preservation. The results were as follows; 1. The enzyme values in reperfusate were highest at 5 minute and after then declined. 2. At 5 minutes after reperfusion, the enzyme values in Group I were lower than those in other Groups. These results suggest that the cardioplegic solutions using for cardiac arrest and myocardial protection can be working better at 4oC than at any other temperature.

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The Clinical Experience of 610 Cases Open Heart Surgery (개심술 610례에 관한 임상적 고찰)

  • 정황규
    • Journal of Chest Surgery
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    • v.21 no.1
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    • pp.36-47
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    • 1988
  • 610 cases of open heart surgery was performed in the Department of Thoracic and Cardiovascular Surgery, Pusan National University Hospital from July 1981 to September 1987. The clinical data was summarized as follows: 1. The age distribution of congenital heart surgery patients was 2 to 41 years old [mean; 13.2 years] and surgery for acquired heart disease was 10 to 57 years old [mean: 32.8 years]. 2. There were 389 cases [63.8%] of acyanotic congenital heart anomalies, 63 cases [10.3%] of cyanotic congenital anomalies and 158 cases [25.9%] of acquired heart disease. 3. For myocardial protection, Bretschneider and potassium glucose solution had been used as cardioplegic solution and then since 1983, GIK solution has been used with repeated infusion method once for every 20 to 30 minutes of time interval after starting initial cardioplegia during operation with excellent results. 4. The ingredient of the priming solution is Hartmann`s solution, sodium bicarbonate, mannitol, potassium chloride, fresh ACD whole blood, calcium chloride, heparin and dexamethasone. 5. There were 96 cases [15.7%] of mild hypothermia, 333 cases [54.6%] of moderate hypothermia and 181 cases [29.7%] of intermediate hypothermia. 6. The mortality rate was 2.3% [9 out of 389 cases] in acyanotic congenital heart disease, 36.5% [23 out of 63 cases] in cyanotic congenital heart disease and 10.8% [17 out of 158 cases] in acquired heart disease, with overall mortality rate of 8.0% [49 out of 610 cases].

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