• Title/Summary/Keyword: 용혈

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Characteristics of the Hemolytic Factor in the Body Fluid from Redworm, Lumbricus rubellus (지렁이 (Redworm*: Lumbricus ruellus) 체액내 용혈인자의 특성)

  • 손영종;이정우장정순
    • The Korean Journal of Zoology
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    • v.34 no.1
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    • pp.20-30
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    • 1991
  • 지렁이(L. rubelIus) 체액내의 적혈구용혈능깍 단백질분해능을 확인하였다. 지렁이의 체액 0. 33 $\mu$l속에 있는 용혈인자는 9 $\times$ 1011 rat RBCs를 2분만에 완전히 용혈시켰으며, 다른 포유류들의 적혈구에도 약간의 차이는 있으나 비슷한 용혈능을 보였고, 이 용혈인자는 지렁이의 혈구가 아닌 몸체조직에서 분비되는 것으로 생각되었다. 용혈인자는 pH 6.5-7.5사이에서 활성이 가장 강하였고, 63'c로 30분 열처리할 경우 활성이 완전히 없어졌으며, 2-mercaptoethanol은 용혈인자의 활성을 증가시켰다. 이 인자의 활성도는 여러가지의 당류, LPS, cholesterol, prosphatidyl Choline, Ch10ropromazine, Sphin90myelin 및 FG2+, Fe3+, CU2+, ZR2+ 등의 금속이온에 의하여 활성이 저해되었다. 한편 지렁이 체액내의 단백질 분해인자는 BSA와 19G를 여러 조각으로 분해시켰으며 이 분해인자는 PMSF및 TLCK에 의하여 활성이 억제되지 않았다. 지렁이의 체강에는 용혈소를 분비하는 박테리아들이 존재하였으나 이들 박테리아들의 용혈소는 지렁이의 용혈인자와는 전기영동이동도에서 차이가 있었다.

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Purification and Characterization of the Extracellular Hemolysin of Vibrio furnissii (Vibrio furnissii로부터 세포외 용혈소의 정제 및 특성)

  • HEO Moon-Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.34 no.5
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    • pp.502-508
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    • 2001
  • The extracellualr hemolysin produced by pathogenic Vibrio furnissii was isolated and purified by ammonium sulfate precipitation and followed by DEAE-Sephadex A-50 (1st and 2nd) ion-exchange and Sephadex G-100 gel permeation chromatography. The molecular weight of the purified hemolysin was 61 kDa. The enzyme activity was inactivated by heating at 60 for 5 min, and inhibited by additions of $Cu^{2+},\;Zn^{2+}$ and a high concentration of cholesterol. Lysis of human erythrocytes by the enzyme was temperature dependent, and optimal temperature of the enzyme was $37^{\circ}C$. The purified hemolysin was unstable at pH 6.0, 6.5 and 10.0. The optimal concentration of human erythrocytes on hemolysins was about $1.5\%$. The purified hemolysin was active for erythrocytes from three animal species including mouse, rabbit and rat, and was the most active against rabbit erythrocytes. B blood group of human erythrocytes also showed a high sensitivity to the enzyme.

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Harmonization of Hemolysis Index in Clinical Chemistry Laboratory and Its Application as a Result Verification Tool (화학검사 결과의 검증을 위한 용혈 지수 일치화 도구 개발)

  • Pyo, Sang Shin;Nam, Hyun Su;Cha, Young Jong;Lee, Seungkwan;Lee, Hae Kyung
    • Korean Journal of Clinical Laboratory Science
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    • v.49 no.4
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    • pp.350-358
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    • 2017
  • The hemolysis index (HI) is semi-quantitative marker for hemolysis. Because the characteristics of the HI vary from one commercial platform to another, no standardization or harmonization of the HI is currently available. Specimens (N=40) randomly selected from clinical patients were artificially hemolyzed in vitro. The serum of the specimens was then diluted with a 20 mg/dL difference between 0~300 mg/dL based on serum hemoglobin measured using the XE-2100 hematology automation equipment (Sysmex, Japan). Diluted serum was measured using the Hitachi-7600 biochemical automation equipment (Hitachi, Japan) to differentiate between HI and serum hemoglobin. The data showed linearity between HI and serum hemoglobin and that HI 1 contained approximately 20 mg/dL of serum hemoglobin. To determine the blood rejection threshold, the HI was divided into three groups: HI 0~1, HI 4~6, HI 9~15. After another batch of clinical specimens (N=40) was measured using a Hitachi-7600 (Hitachi, Japan), each specimen was moved forward and backward with the piston of the syringe to induce an artificial in vitro hemolysis, then measured again with a Hitachi-7600 (Hitachi, Japan). The percentage difference between the three groups was analyzed by ANOVA or the Kruskal-Wallis test. In the post-test, there were significant differences between the HI 0~1 and the HI 5~6: Glucose, creatinine, total protein, AST, direct bilirubin, uric acid, phosphorus, triglyceride, LDH, CPK, Magnesium, and potassium levels. Because many clinical tests differed significantly, the threshold for hemolysis could be appropriate for HI 5 (serum hemoglobin 100 mg/dL).

The Effect of in vitro Hemolysis on Vitamin $B_{12}$ / Folic Acid Results (용혈이 Vitamin $B_{12}$ / Folic acid 검사 결과에 미치는 영향)

  • Kim, Mi-Sook;Cho, Youn-Kyo;Im, Do-Hw;Seo, So-Yeon
    • The Korean Journal of Nuclear Medicine Technology
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    • v.12 no.1
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    • pp.62-65
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    • 2008
  • Purpose: Vitamin $B_{12}$ and folic acid are type of water-soluble vitamin and those work as a secondary vital enzyme, but especially those are involved in the nuclear DNA synthesis. Those are mainly measured in order to diagnose megaloblastic anemia and to assess the storage of folic acid during pregnancy. It is generally known that the hemolyzed serum is useless for folic acid and vitamin $B_{12}$, but it is not easy to abide by this information because our lab (Samkwang Medical Laboratories) is reference laboratory. We tested how much the extent of hemolyzed serum had influence on the results. Materials and Method: We performed the tests of vitamin $B_{12}$, folic acid. For the hemolysis effect study, we used 40 cases. According to the level of A, B and C groups, each group had 10 samples which were mechanically hemolyzed serum. Results: Hemolysis did not affect the vitamin $B_{12}$ results. However in case of folic acid, the value increased according to the degree of hemolysis. And severe hemolyzed cases had the highest value (greater than 20 ng/ml). Conclusions: Preventing the hemolysis, it has to be informed that blood-collecting, separation and storage are performed more carefully. As you see from the above results, hemolyzed serum is not proper for folic acid test, and hemolysis does not affect the results of vitamin $B_{12}$. However, for the more accurate results, it is necessary to avoid hemolysis.

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A STUDY ON THE HEMOLYTIC PROPERTIES OF PREVOTELLA NIGRESCENS (Prevotella nigrescens의 용혈특성에 관한 연구)

  • Kwak, Ju-Seok;Jang, Hoon-Sang;Jang, Seok-Woo;Lee, Su-Jong;Yu, Yong-Wook;Min, Kyung-San
    • Restorative Dentistry and Endodontics
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    • v.30 no.4
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    • pp.335-343
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    • 2005
  • Hemolytic property is a specific feature of bacteria to obtain iron which is essential for its survival in host tissues. Therefore, it is thought to be one of several factors of virulence. The purpose of this study was to investigate the hemolytic properties of Prevotella nigrescens isolated from the teeth diagnosed as pulp necrosis and apical periodontitis under the presence of hemolysin inhibitors such as $NaN_3$ and dithiothreitol. heat, various pH and cultural conditions. The results were as follows; 1. Clinically isolated P. nigrescens strains and standard P. nigrscens ATCC 33563 showed hemolytic activity. 2. P. nigrescens showed higher hemolytic activity against human erythrocytes than sheep or horse erythrocytes. 3. $NaN_3$ and dithiothreitol (DTT) reduced the hemolytic activity of P. nigrescens in a dose dependent manner (p<0.05). 4. Optimal pH for the maximum hemolytic activity of P. nigrescens was 4.0 and the hemolysin was stable under the $50^{\circ}C$, but the hemolytic activity was significantly decreased at $95^{\circ}C$. 5. P. nigrescens cultured in $10\%\;CO_2$ condition showed higher hemolytic activity than the bacteria cultured in the anaerobic condition.

A Study on Hemolysis Characteristics of Intra-Cardiac Axial Flow Blood Pump (심장내 이식형 축류 혈액펌프 용혈특성에 관한 연구)

  • 김동욱
    • Journal of Biomedical Engineering Research
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    • v.21 no.4
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    • pp.353-362
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    • 2000
  • Minimization of hemolysis is one of the key factors for successful axial flow blood pumps. It is, however, difficult to estimate the hemolytic performance of axial flow blood pumps without experiments. Instead, the Computational Fluid Dynamics(CFD) analysis enables the prediction of hemolysis. Three-dimensional fluid dynamics of axial flow pumps with different impellers were analyzed using the CFD software, FLOTRAN. The turbulence model k-$\varepsilon$ was used. The changes in turbulent kinetic energy applied to each particle (red blood cell) flowing through the pumps were computed and displayed by the particle trace method (particle spacing of 10 msec). Also, the Reynolds shear stress was calculated from the turbulent kinetic energy. The shear stress was higher behind the impellers than elsewhere. The CFD analysis could predict in vitro results of hemolysis and also the areas where hemolysis occurred. The CFD analysis was found to be a useful tool for designing less hemolytic rotary blood pumps.

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Purification of Hemolysin from Vibrio anguillarum Isolated from Fish (어류분리 Vibrio anguillarum 용혈소의 정제)

  • 김영희
    • Journal of Life Science
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    • v.8 no.5
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    • pp.598-603
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    • 1998
  • A marine microbe, Vibrio anguillarum was isolated from fish and studied for its concerning pathogenic substance of hemolysin. Purification of hemolysin was achieved by the procedure of ammonium sulfate precipitation from cul-ture filtrate, DEAE-cellulose chromatography, and G-200 gel filtration with 36 fold of purification and 2.3% yield. The molecular weight of the purified hemolysin was 38,000 dalton by SDS-PAGE. The purified hemolysin was stable at pH 6-9, below 45$^{\circ}C$, and up to 1% of NaCl, respectively. $Ca^{2+}, Cu^{2+}, Zn^{2+}, Fe^{2+}$ inhibited the hemolytic activity whereas EDTA and $Mg^{2+}$ did not.

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Studies on the Hemolytic Activities of Korean Wild Mushrooms (II) - Screening of 22 Mushrooms Including Laccaria vinaceoavellanea for Their Hemolytic Activities - (한국산 야생버섯의 용혈작용에 대한 연구 -제 2보: 색시졸각버섯(Laccaria vinaceoavellanea)등 22종의 in vitro 용혈활성 검색 -)

  • Chung, Kyeong-Soo;Lee, Ji-Seon;Jo, Moon-Joo;Lee, Im-Seon
    • The Korean Journal of Mycology
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    • v.28 no.2
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    • pp.123-125
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    • 2000
  • To investigate the hemolytic toxicities of the Korean basidiomycetes, the cold-water extracts of 22 wild mushrooms were tested for their hemolytic activities on the erythrocytes of a mouse and a rat. As a result, three species including Collybia confluence, Laccaria vinaceoavellanea and Lenzites betulina showed strong hemolytic activities. Of these, L. vinaceoavellanea retained more than 80% of its hemolytic activity even after it was boiled for 30 minutes, while the other two were inactivited in five minutes.

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Studies on the Hemolytic Activities of Korean Wild Mushrooms (III) - Hemolytic Characteristics and in vivo Toxicity of Hemolysin of Hebelma crustuliniforme - (한국산 야생버섯의 용혈작용에 대한 연구 - 제 3보 : 무우자갈버섯(Hebeloma crustuliniforme) 용혈독소의 용혈특성 및 in vivo 독성 -)

  • Yang, Hee-Jung;Lee, Ji-Seon;Chung, Kyeong-Soo
    • The Korean Journal of Mycology
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    • v.30 no.2
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    • pp.119-123
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    • 2002
  • In the previous studies, we surveyed 68 Korean wild mushrooms for their hemolytic activity and found that cold-water extract of Hebeloma crustuliniforme contained heat-resistant hemolysin. In this study, partially purified hemolysin of the mushroom was obtained by cold-water extraction followed by precipitation with ammonium sulfate, solubility fractionation and then dialysis. The hemolysin was found to be > 12,000 in molecular weight and its optimal hemolytic temperature was $37^{\circ}C$ and it's hemolytic activity, on washed erythrocytes and unwashed erythrocytes, respectively, was in the order of sheep > rat > human ${\geq}$ mouse > chicken and sheep > mouse > human ${\geq}$ rat > chicken. When ip injected into ICR mice at 1.38 mg/kg, it incurred prompt hemolysis as well as severe renal toxicity and hepatotoxicity. These results strongly suggest that the toxicity of Hebeloma crustuliniforme, which had been well-known as a toxic wild mushroom, may be at least partly due to its hemolysin.

The Effect of Hemolysis sample on the Result of Nuclear Medicine Blood test (용혈검체가 핵의학 검체검사 결과에 미치는 영향)

  • Kim, Jin-Tae;Lee, Jong-Pil;Lee, Soo-Bin;Kim, Dong-Min
    • The Korean Journal of Nuclear Medicine Technology
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    • v.25 no.1
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    • pp.41-43
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    • 2021
  • Purpose In nuclear medicine blood tests, hemolysis samples are considered as inappropriate sample and are recommended not to be used for blood test. So, the lab are required to collect the blood again in the blood collection room However, The effect of hemolyzed samples on radioimmunoassay has not studied yet. This study was designed to evaluate effects of hemolysis on radioimmunoassay. Materials and Methods The kit manuals of 23 test items were reviewed to confirm whether hemolyzed samples were used. The subjects were 19 general applicants(male : 9, female : 13) and the samples were collected by each two SST tubes, one tube was obtained by centrifugation normally, and the other was obtained hemolyzed sample by centrifugation after external shock. It has been known that highly hemolyzed samples can affect the test results, so the test was performed using the severe hemolyzed sample. The test was performed for each test item using 23 normal serum and hemolysis serum, and SPSS19 program was used for statistical comparison of the test result. Results There was no significant difference between normal serum and hemolysis serum in 21 of 23 test items, but the results of insulin and C-peptide were significantly different(P<0.05). Conclusion It has been known that hemolysis in blood samples can affect the results of biochemical and hematological test, However, hemolysis effect is relatively low. Similarly, this study showed that hemolysis had not much effect on most of immunological radioimmunoassay except for some tests. Therefore, it is thought that the demand for re-collection due to hemolysis will be reduced in the laboratory, which will improve the work process of the laboratory.