• Title/Summary/Keyword: cell agglutination

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Development of PCR assay for the detection of Brucella spp in bovine semen (종모우 정액중 Brucella균 신속 검출을 위한 PCR기법 개발)

  • Jung, Suck-chan;Jung, Byeong-yeal;Woo, Seong-ryong;Cho, Dong-hee;Kim, Jong-yeom;Kim, Woo-taek;Lee, Jung-mi;Park, Yong-ho;Baek, Byeong-kirl
    • Korean Journal of Veterinary Research
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    • v.38 no.2
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    • pp.345-352
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    • 1998
  • The diagnosis of brucellosis is currently based on serological and microbiological tests. However, the microbiological isolation and identification have several disadvantages such as time-consuming and laborious, and the serological methods have been reported to cross-react with antigens other than those from Brucella spp. To develop a sensitive and rapid diagnostic method for detection of Brucella species, the genus-specific primers were designed and synthesized from the sequence of gene encoding a 31kDa cell surface protein(BCSP) and a 36kDa outer membrane protein(OMPB) of B abortus. The amplified 711bp and 982bp DNA fragments were only visible in each species of Brucella by PCR method using the BCSP and OMPB primers, respectively. However, PCR product was not obtained with DNA from other Gram-negative bacteria. As little as 1pg of the B abortus genomic DNA could be detected by this PCR method. Using the PCR technique, semen samples from 185 bulls of Brucella-seronegative herds in Cheju island were examined for comparison of this PCR method with conventional methods in 1995. The semen samples from 5 bulls were positive by culture method and PCR, and one was positive and 5 were suspect by semen plasma agglutination test. However, the semen samples obtained from 177 bulls were negative by semen plasma agglutination, culture and PCR methods in 1996. The results of comparison tests suggested that PCR was a better test than agglutination test against semen of bulls. This study indicated that the PCR technique was a valuable for the diagnosis of bovine brucellosis, particulary in bull semens.

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Studies on Lectins from Marine Shells (III) : Screening of Lectin-like Agglutinins from Marine Shells

  • Chung, See-Ryun;Kim, Jang-Hwan;Suh, Young-Ah;Jeunechung, Kyung-Hee
    • Archives of Pharmacal Research
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    • v.9 no.4
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    • pp.201-203
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    • 1986
  • Forty species of marine shells were collected from Korean coasts and studied extensively for their lectin activities by using erythrocytes of human blooc A, AB, B. O group and rabbit blood. In total, 7 species contained lectines :Neptunea intersculpta, Omphalius nigerrimus and Scapharca subcrenata, blood group nonspecific; Saxidomus purpuratus, human blood A and AB group specific; Lepidozona coreana, Tegilarca granosa and Neptunea polycosta, rabbit blood specific.

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Studies on Lectins from Marine Animal Chlorostoma argyrostoma turbinatum (해양동물 구멍밤고둥의 렉틴 성분 연구)

  • Chung, See-Ryun;Choi, Il-Shik;Jeune, Kyung-Hee
    • Korean Journal of Pharmacognosy
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    • v.25 no.2
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    • pp.121-131
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    • 1994
  • Two kinds of new lectins, CATL-I and CATL-II, were partially purified from the intestine of Chlorostoma argyrostoma turbunatum by physical saline extraction, salt fractionation, ion exchange chromatography and gel filtration. CATL-I and CATL-II were purified 39.4 and 15.8 fold with a yield of 8.8 and 7.4%, respectively. On polyacrylamide gel electrophoresis, CATL-I demonstrated one major and one minor bands. This lectin agglutinated human and other animal erythrocytes nonspecifically and also agglutinated murine splenic lymphocytes. Carbohydrate specificity of the lectins was determined by inhibition of the agglutinability by methyl-${\alpha}-_D$-galactopyranoside and $_L-rhamnose$ at a final concentration of 6 mM. The lectins contained relatively high amounts of acidic amino acids, but the contents of sulfur containing amino acids were very low or was not estimated. Immunochemical studies were carried out to identify some properties of marine animal lectins.

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Electrophoretical Analysis of 36-Kilodalton Outer Membrane Protein of Vibrio vulnificus ATCC 27562

  • Moon-Soo Heo;Cho-Rok Jung
    • Journal of Life Science
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    • v.9 no.1
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    • pp.35-39
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    • 1999
  • Elecrophoreticl analysis of a 36 kDa protein was runned by SDS-PAGE, isoelectric focusing (IEF) and two dimensional electrophoresis pattern. Major 36 kDa and 25, 46, 48, 66 kDa protein were detected by Coomassie blue stain on SDS-PAGE. Major 36kDa protein was eluted for production of antiserum for serological analysis, IEF and two dimensional electrophoresis. Isoelectric point of 36kDa was aout pH 8.5. Two dimensional electrophoresis of eluted 36kDa showed one point on the gel. Anti-36 kDa serum made by newzilland rabbit for serological test. In ELISA, final titer of antibody was 100×{TEX}$2^5}${/TEX} : 1. Neutralize ability of serum was examined by slide agglutination test and colonization test in rat. Anti-36 kDa serum agglutinated whole cell of V. vulnificus were inhibited colonization on intestine in rat. Accordingly In this paper contain some electrophoretical analysis and serological test of a 36 kDa OMP of V. vulnificus.

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Immune-mediated hemolysis after administration of human intravenous immunoglobulin in a dog: a case report

  • Minji Kim;Youngju Kim;Hyeona Bae;Rankyung Jung;Minjeong Kang;Sumin Cha;Kyu-Woan Cho;Dong-In Jung;DoHyeon Yu
    • Korean Journal of Veterinary Research
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    • v.63 no.3
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    • pp.28.1-28.5
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    • 2023
  • A 10-year-old spayed female Maltese presented with purpura and hematemesis. Initial laboratory evaluation revealed immune-mediated thrombocytopenia, but evidence of hemolytic anemia was not identified. Three milligrams of human intravenous immunoglobulin (hIVIG) was administered for 3 hours following prednisolone and mycophenolate mofetil. A pale mucous membrane was identified, and the packed cell volume decreased by 3%. Blood film examination revealed significant spherocytosis with auto-agglutination. Blood transfusions and immunosuppression were continued for 4 days, and hIVIG was discontinued. This report describes a case of increased immune-mediated hemolysis after hIVIG administration, possibly due to new-onset immune-mediated hemolytic anemia or enhanced immunogenicity.

The Frequency and Distribution of Unexpected Red Cell Antibodies and Analysis of Antigen Exposure

  • Yang, Sung Sik;Yang, Hoi Joo;Park, Hyun Jun;Kwon, Seog Woon
    • Korean Journal of Clinical Laboratory Science
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    • v.45 no.4
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    • pp.164-169
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    • 2013
  • Red cell alloantibodies other than naturally occurring anti-A or anti-B are called unexpected red cell antibodies, and can be detected by performing an antibody screening. The frequency and distribution of unexpected antibody identified in Asan Medical Center were analyzed. We investigated a total of 135,238 cases of antibody screening test in AMC for 3 years from 2010 to 2012. Using column agglutination techniques, antibody identification tests were performed for the cases with positive antibody screening. Among 135,238 cases, 854 (0.6%) cases showed positive results of antibody screening test. In the order of frequency, 284 (33.3%) anti-Rh, 89 (10.4%) anti-MNS, 62 (7.3%) anti-Lewis, 34 (4.0%) anti-Kidd, 10 (1.2%) anti-Duffy, and 9 (1.1%) anti-P were identified. Multiple antibodies were detected in 199 (23.3%) cases. Among 381 subjects investigated for transfusion history, 299 (78.5%) had history of transfusion while 82 (21.5%) had unknown history. Thus the incidence of unexpected antibody was higher in the group with history of transfusion than the group without (p<0.001). Also, among 435 subjects investigated for the history of pregnancy, 46 (10.6%) had no history while 389 (89.4%) had history of pregnancy, showing higher incidence of unexpected antibody in the group with history of pregnancy than the group without pregnancy (p<0.001). Evaluated amounts and frequency of antigen exposure due to transfusion and pregnancy is suggested to increase the frequency of identification of unexpected antibody.

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Characteristics and Cancerostatic Activity of the Starfish Lectin (별불가사리 렉틴의 특성 및 암 세포 성장저해 효과)

  • Jeune, Kyung-Hee;Park, Chae-Soo;Park, Won-Hark;Choi, Soo-Jeong;So, Myung-Suk;Chung, See-Ryun
    • YAKHAK HOEJI
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    • v.41 no.4
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    • pp.421-432
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    • 1997
  • A new lectin was partially purified from starfish,Asterina pectinifera by means of physiological saline extraction, salt fractionation, ion exchange chromatography and hy droxyapatite chromatography, and it was named APL. The biochemical properties of the APL were characterized. In addition, its effects on lymphocyte mitogenicity and cancer cell agglutinability were tested. The APL agglutinated nonspecifically human erythrocytes and rabbit blood cells. Agglutinability was decreased to 30% of control activity below pH 5 and above pH 9 and was relatively unstable at increasing temperatures above 60$^{\circ}C$. The activity was reduced by addition of two kinds of metal ions, $Ba^{2+},\;Mn^{2+}$ and chelating agent, EDTA. APL was proved to be glycoproteins containing 9% sugars. For carbohydrate specificity, it was found that the activity of APL was inhibited by D(+)-glucosamine, D(+)-galactosamine, stachyose, N-acetyl-galactosamine and methyl-${\alpha}$-D-galactopyranoside among 35 sugars tested. In amino acid composition, the contents of acidic amino acids such as aspartic acid and glutamic acid were relatively high. This result suggest that the isoelectric point would be in a lower range. APL was found that it promotes the division of human lymphocytes. APL was proved to be a potent agglutinin for cancer cells such as HeLa, L929 and L1210 cells. Significant changes on the HeLa cell surfaces affected by APL were observed under the electron microscope.

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Development and Immunochemical Properties of Two Monoclonal Antibodies Specific to Human Chorionic Gonadotropin

  • Kim, You-Hee;Koh, Kwan-Sam
    • BMB Reports
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    • v.32 no.5
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    • pp.474-479
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    • 1999
  • Using a hybridoma technique, spleen cells of Balb/c mice immunized with human chorionic gonadotropin (hCG) were fused with NS-1 mouse myeloma cells. Two hybrid cell lines, clones KS-8 and KS-19, secreting monoclonal antibodies to hCG, were isolated. KS-8 and KS-19 belong to the immunoglobulin $G_1$ subclass. With the aid of a double-antibody radioimmunoassay, it was established that the KS-8 monoclonal antibody recognizes an immunodeterminant of the $\beta$-subunit of hCG, whereas the KS-19 monoclonal antibody recognizes an epitope present on the $\alpha$-subunit of hCG. The KS-8 monoclonal antibody specifically reacts with human chorionic gonadotropin and shows cross-reactivity of less than 0.3% to other related human glycoprotein hormones. On the other hand, using a hemagglutination test based on antibody-induced agglutination of sheep red blood cells coated with hCG, It was shown that only the KS-19 monoclonal antibody was capable of inducing a positive reaction, although both monoclonal antibodies had similar binding capacity to the coated cells. The results from the dual screening procedures demonstrate that KS-8 and KS-19 monoclonal antibodies show high sensitivity in two different assays, and are hence useful for the qualitative and quantitative determination of hCG by both radioimmunoassay and hemagglutination inhibition tests.

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Purification and Characterization of a New Galactoside Specific Lectin from Trichosanthes kirilowii Root

  • Yun, Doo-Hee;Park, Eun-Ju;Park, Jong-Ok;Lee, Young-Han;Seo, Jeong-Kon;Ryu, Sung-Ho;Suh, Pann-Ghill;Kim, Hee-Sook
    • BMB Reports
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    • v.28 no.1
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    • pp.6-11
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    • 1995
  • A new lectin, named TRA, was purified from Trichosanthes kirilowii root by acid-treated Sepharose 6B, Mono-Q, and TSK-gel 3000SW column sequential chromatography. The lectin appeared homogeneous by native gel electrophoresis at pH 4.3 and gave two protein bands of Mr=31 and 28 kDa by SDS-PAGE. The N-terminal amino acid sequences of the polypeptides of TRA have not been reported in amino acid sequences of the lectins. TRA lectin formed a precipitate with asialofetuin, neuraminidase-treated fetuin. A sugar inhibition assay indicated that N-acetyl-D-galactosamine, among the monosaccharides tested, was the most potent inhibitor of TRA-induced hemagglutination. Asialofetuin showed a 260-times stronger inhibitory activity than N-acetyl-D-galactosamine. TRA lectin also showed agglutination with normal leukocytes and lymphoma cells, but not with premature hemopoietic cells. These results suggest that TRA is a novel plant lectin.

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Effect of Lavender Oil on the Ultraviolet-Damaged Mice Skin (자외선으로 손상된 마우스 피부에 대한 라벤더 오일 효과)

  • Rhie, Sung-Ja;Sim, Mi-Ja;Kim, Young-Chul
    • Environmental Analysis Health and Toxicology
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    • v.23 no.4
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    • pp.315-323
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    • 2008
  • In this study, the backs with a hair cut of 6-week-old healthy ICR male mice were once exposed to a dose of $400\;mJ/cm^2$ UVB. An acute dermal inflammation was observed, and the certified 100% pure and natural lavender essential oil were applied to the UVB-exposed mice skin twice a day. It was observed that the mice exposed to UVB resulted in an acute inflammation, and when treated with lavender oil the degree of inflammation was much alleviated, and the inflamed skins of both the control and lavender oil-treatment groups were cured almost completely after 6 days of the UVB exposure. At 24 hours after UVB exposure, the epidermal keratinocytes in the control group showed a cell-membrane damage with the destruction of intercellular junctions, agglutination of tonofilaments within the cytoplasm and nucleus damage, while the lavender oil-treatment group had much less cell damage than the control group. While the control group showed a significant increase (p<0.05) in the activity of XO up to 144 hours, the lavender oil-treatment group did not show any significant increase except for 48 hours after the UVB exposure. Both the control and lavender essential oil-treatment groups had a significant decrease in the activities of CAT and SOD up to 96 hours. Particularly, the CAT activity was significantly lower(p<0.05) in the lavender oil-treatment group than the control group up to 48 hours, and higher than the control group at and after 96 hours. The GST activity was significantly decreased in both the control and lavender oil-treatment groups up to 96 hours after the UVB exposure except for the control group at 24 hours, and that of the lavender oil-treatment group was higher than the control group at and after 96 hours. Therefore, it is assumed that the application of the lavender oil to the ultraviolet-damaged mice skin can be effective in treatment for the damaged skin.