• Title/Summary/Keyword: Crude antigen

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A Surge in the Seroprevalence of Toxoplasmosis among the Residents of Islands in Gangwha-gun, Incheon, Korea

  • Yang, Zhaoshou;Cho, Pyo-Yun;Ahn, Seong-Kyu;Ahn, Hye-Jin;Kim, Tong-Soo;Chong, Chom-Kyu;Hong, Sung-Jong;Cha, Seok-Ho;Nam, Ho-Woo
    • Parasites, Hosts and Diseases
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    • v.50 no.3
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    • pp.191-197
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    • 2012
  • Seroepidemiological changes of Toxoplasma gondii infection among the residents of the islands of Gangwha-gun, Incheon for 2 years were surveyed and evaluated by ELISA using a crude extract antigen. In 2010, sera of 919 adult residents in Gyodong-myeon and 313 adults in Samsan-myeon were collected and checked for IgG antibody titers, which showed 14.5% (133 sera) and 19.8% (62 sera) positive rates, respectively. In 2011, sera of 955 adults in Gyodong-myeon and 341 adults in Samsan-myeon were examined, which showed an increase of positive rates to 23.8% (227 sera) and 31.7% (108 sera), respectively. Totally, the seroprevalence of the first year was 15.8% and it increased rapidly to 25.8% in the second year. The positive rates of both sexes increased simultaneously with the significant ratio of males to females by 1.7-2.2 fold (P<0.05). In both myeons, 661 sera were collected every year and showed changes in optical density (OD) in 177 sera; newly found as positives in 73 persons (11.0%), negative conversion in 10 persons (1.5%), and maintained or increased in 94 persons (14.2%). This rapid increase in the prevalence of toxoplasmosis in Gangwha islands may be due to in part peculiar changes in the toxoplasmic environment of the islands and presumably the consumption of the pork bred domestically within the islands or imported from high endemic nations. It is necessary to find out symptomatic toxoplasmic patients and confirm the risk factors for further infection in the islands of Gangwha-gun.

Immunoblot observation of antigenic protein fractions in Paragonimus tvestermani reacting with humall patients sera (폐흡충 항원단백질에 대한 폐흠충증 한자 혈청의 반응 양상)

  • Kim, Sung-Hwan;Kong, Yoon;Kim, Suk-Il;Kang, Shin-Yong;Cho, Seung-Yull
    • Parasites, Hosts and Diseases
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    • v.26 no.4
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    • pp.239-244
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    • 1988
  • In order to observe the antigenic fractions in saline extract of adult Paragonimus westermani, proteins in the crude extract were separated by sodium dodecyl sulfate-polyacylamide gel electrophoresis(SDS.PAGE) in reducing conditions. The separated protein fractions were transferred to nitrocellulose paper on which 20 sera from human paragonimiasis were reacted and immunoblottrd. Out of 15 stained protein bands in SDS-PAGE, 7 reacted with infected sera while 8 did not. Additionally, 7 unstained protein bands in SDS-PAGE reacted with the sera. Of 14 reacted bands, 30 kilodalton(kDa) band was the most frequently reacted (95%) and was a strong antigen. Protein bands of 23 and 46 kDa were also strong antigens. Bands of over 150 kDa, 120 kDa, 92 kDa, 86 kDa, 74 kDa, 62 kDa, 51 kDa, 32 kDa, 28 kDa, 16.5 kDa and 15.5 kDa were also reactive but their frequencies of the reaction were variable. Key words: Paragonimus westermani, human paragonimiasis, antigenic proteins, SDS-PAGE/ immunoblot

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Variation of antigenic proteins of eggs and developmental stages of Paragonimus westermani (폐흡충 발육 단계에 따른 항원 단백질의 변화)

  • Yoon KONG;Joon-Yong CHUNG;Doo-Hee YUN;Lee-Su KIM;Shin-Yong KANG;Akira ITO;Liang MA;Seung-Yull CHO
    • Parasites, Hosts and Diseases
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    • v.35 no.3
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    • pp.197-202
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    • 1997
  • Diagnosis of early paragonimiasis is difficult because parasitological evidence is not easily obtained. Antibody tests have been proposed as a good substitute for classical diagnostic techniques. Using the crude extracts of Parnsonimus westermnni eggs, metacercariae. 4- and 7-week Juveniles, and 16-week adults as antigens, we observed the early antibody responses. Sera were obtained from 4 experimental cats, fed 50 metacercariae each, at intervals until 13 weeks post-infection. Antibody (IgG) responses were identified by ELISA using extracts of 4-week juveniles, followed by those of 7- and 16-week worms. Antibody responses were minimal against the metacercarial extracts. Antibodies to p. westemoni egg extracts were elevated after 10 weeks post-infection. In immunoblot analysis, more than nine protein bands in 4-week juveniles reacted with the early infection sera. Antigenic proteins in adult worms were different from those of juveniles. After four weeks of infection, 32 and 35 kDa bands in the adult extracts were increasingly reactive. Egg specific proteins at 28, 46 and 94 kDa were reactive only after 10 weeks. Antigenic components reactin료 to the early infection sera changed during the maturation stages of P. westermani; almost all juvenile antigens were replaced by adult antigen components.

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Characterization of the Immunologically Active Components of Glycyrrhiza uralensis Prepared as Herbal Kimchi

  • Hwang, Jong-Hyun;Lee, Kyong-Haeng;Yu, Kwang-Won
    • Preventive Nutrition and Food Science
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    • v.8 no.1
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    • pp.29-35
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    • 2003
  • A crude polysaccharide fraction (GU-3) from the roots of Glycyrrhiza uralensis (licorice root), a screened herbal plant used in the preparation of herbal kimchi, enhanced Peyer's patch mediated bone marrow cell proliferation and NK cell-mediated tumor cytotoxicity against Yac-1 cells. GU-3 was further purified by DEAE-Sepharose CL-6B yielding fractions designated as GU-3I, and 3IIa∼3IIe. GU-3IIa is mainly composed of arabinose, galactose and galacturonic acid, and showed the highest bone marrow cell proliferation activity. In addition, GU-3IIb had arabinose, galactose, rhamnose and galacturonic acid as the component sugars with a small quantity of protein; GU-3IIb also enhanced activity of NK cell-mediated tumor cytotoxicity. After these fractions were further fractionated via gel filtration on Sepharose CL-6B or Sephacryl S-300, two immunological active polysaccharides, GU-3IIa-2 and 3IIb-1 were purified from the respective fractions. GU-3IIa-2 mostly contained neutral sugars (75%) such as arabinose and galactose (molar ratio; 1.0 : 0.7) in addition to a considerable amount of galacturonic acid (20%), whereas GU-3IIb-1 was composed of arabinose, galactose, rhamnose and galacturonic acid (molar ratio; 0.3 : 0.5 : 0.1 : 1.0). Methylation analysis indicated that GU-3IIa-2 was composed mainly of terminal, 4- or 5-linked and 3,4- or 3,5-branched arabinose, 3-linked, 4-linked and 3,6-branched galactose, and terminal and 4-linked galacturonic acid whereas GU-3IIb-1 contained various glycosidic linkages such as terminal and 4- or 5-linked arabinose, 2,4-branched rhamnose, terminal and 4-linked galactose, and terminal and 4-galacturonic arid. Single radial gel diffusion indicated that only GU-3IIa-2 strongly reacted with β-D-glucosyl-Yariv antigen. These results suggest that bone marrow cell proliferating activity and enhancement of NK cell-mediated tumor cytotoxicity of GU-3 are caused by polysaccharides containing a pectic arabinogalactan (GU-3IIa-2) and pectic polysaccharide (GU-3IIb-1).

Purification of antigenic proteins of Paragonimus westermani and their applicability to experimental cat paragonimiasis (폐(肺)디스토마(Paragonimus westermani) 감염(感染) 고양이 혈청(血淸)에 대(對)한 ELISA 항체가(抗體價)의 의의(意義))

  • Choi, Won-Young;Yoo, Jae-Eul;Nam, Ho-Woo;Choi, Hyung-Rak
    • Parasites, Hosts and Diseases
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    • v.24 no.2
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    • pp.177-186
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    • 1986
  • This study was designed to evaluate the partially purified antigens which were fractionated from crude extract of Paragonimus westermani and to monitor the enzyme-linked immunosorbent assay (ELISA) in experimental cat paragonimiasis during the course of infection as well as before and after chemotherapy. Crude extract of 6-month-old adult P. westermani was fractionated to 5 antigens by successive applications of ammonium sulfate precipitation, ion exchange chromatography and gel filtration. And the cats, 10 in each group, were infected with 60, 30, 15, and 5 metacercariae, then the half of each group was treated with praziquantel 2 times in one day of 100mg per kilogram of weight on 150 days after the infection. Sera were collected every 10 days. ELISA was performed with the concentration of $2{\mu}g/ml$ antigen, 100 times diluted sera and 1,000 times diluted alkaline phosphatase conjugated anti-cat IgG. The results were as follows: 1. Absorbance by ELISA with proteins precipitated by differential concentration of ammonium sulfate was the highest at $51{\sim}65%$ precipitate (PA2), followed by $0{\sim}50%$ precipitate (PAl), $66{\sim}80%$ precipitate (PA3), and $81{\sim}90%$ precipitate (PA4). Unprecipitated protein over 90% ammonium sulfate (PA5) showed the lowest antigenicity. 2. Fractionation of PA1, PA2, and PA3 through the DEAE-cellulose column did not differentiate the antigenic proteins. 3. By passing through the Sephadex G-200 column, PA1 and PA2 were fractionated to high molecular weight proteins and those of low molecular weight which showed high absorbance by ELISA (PA1-I, II and PA2-I, II). But PA3 was shown to have a fraction of high molecular weight proteins (PA3-I) which showed high antigenicity. 4. SDS-polyacrylamide gel electrophoresis of PA1-I, P A1-II, PA2-I, PA2-II, PA3-I, and crude extract was performed. Fraction PA1-I was composed of proteins which had the molecular weight of 270 kilodaltons(KD) to 196 KD; of them 220KD protein was major band. Fraction PA2-I was composed of $255{\sim}225\;KD$, and PA3-I, $255{\sim}240\;KD$, respectively. Fraction PA1-II and fraction PA2-II consisted of 30 KD proteins. 5. Absorbance by ELISA began to increase within $10{\sim}20$ days after the infection and reached the highest on $140{\sim}180$ days, then made plateau thereafter. 6. Absorbance by ELISA decreased after praziquantel treatment. In 60 metacercariae infection group, the absorbance had been decreasing, but remained within the positive range during observation period, while those of 30, 15, and 5 metacercariae infection groups turned to negative range. 7. Fraction PA1-II showed the highest antigenicity in ELISA, then fraction PA2-I, fraction PA1-I, fraction PA2-II, fraction PA3-I and crude extract followed. In early phase of infection, the absorbance of fraction PA1-II showed more rapid increase than those of the other fractions and it came to positive range at $20{\sim}30$ days after infection.

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