• Title/Summary/Keyword: Spirometra mansoni plerocercoid(sparganum)

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Partial Purification and Characterization of a Cysteine Protease Inhibitor from the Plerocercoid of Spirometra erinacei

  • Chung, Young-Bae;Yang, Hyun-Jong
    • Parasites, Hosts and Diseases
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    • v.46 no.3
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    • pp.183-186
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    • 2008
  • Helminthic cysteine proteases are well known to play critical roles in tissue invasion, nutrient uptake, and immune evasion of the parasites. In the same manner, the sparganum, the plerocercoid of Spirometra mansoni, is also known to secrete a large amount of cysteine proteases. However, cysteine protease inhibitors regulating the proteolytic activities of the cysteine protease are poorly illustrated. In this regard, we partially purified an endogenous cysteine protease inhibitor from spargana and characterized its biochemical properties. The cysteine protease inhibitor was purified by sequential chromatographies using Resource Q anion exchanger and Superdex 200 HR gel filtration from crude extracts of spargana. The molecular weight of the purified protein was estimated to be about 11 kD on SDS-PAGE. It was able to inhibit papain and 27 kDa cysteine protease of spargana with the ratio of 25.7% and 49.1%, respectively, while did not inhibit chymotrypsin. This finding suggests that the cysteine protease inhibitor of spargana may be involved in regulation of endogenous cysteine proteases of the parasite, rather than interact with cysteine proteases from their hosts.

Sparganum in a Hen (닭의 Sparganum 감염(感染)에 대(對)하여)

  • Kim, Young Sup;Kim, Dong Hee
    • Korean Journal of Veterinary Research
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    • v.13 no.1
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    • pp.75-77
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    • 1973
  • A large plerocercoid was found from abdominal cavity of hen slaughtered in a slaughter-chicken house at Chyung Yang market in Seoul. The worm was identified as Diphyllobothrium mansoni of Genus Spirometra and the length of whole body was approximately 50 cm.

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Enzymatic N-glycan analysis of 31 kDa molecule in plerocercoid of Spirometra mansoni (sparganum) and its antigenicity after chemical oxidation

  • Chung, Young-Bae;Kong, Yoon;Yang, Hyun-Jong
    • Parasites, Hosts and Diseases
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    • v.42 no.2
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    • pp.57-60
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    • 2004
  • A highly specific antigenic protein of 31 kDa from plerocercoid of Spirometra mansoni (sparganum) was obtained by gelatin affinity and Mono Q anion-exchange column chromatography. The purified 31 kDa protein was subjected to N-glycan enzymatic digestion for structural analysis. The relative electrophoretic mobility was analyzed by SDS-PAGE, before and after digestion. On SDS-PAGE after enzymatic digestion, the 31 kDa protein showed a molecular shift of approximately 2 kDa, which indicated the possession of complex N-linked oligosaccharides (N-glycosidase F sensitive) but not of high-mannose oligosaccharides (endo-beta-N-acetylglucosaminidase H, non-sensitive). Chemically periodated 31 kDa protein showed statistically non-significant changes with human sparganosis sera by enzyme linked immunosorbent assay (ELISA). Therefore, the dominant epitopes of the 31 kDa molecule in human sparganosis were found to be mainly polypeptide, while N-glycans of the antigenic molecule in sparganum was minimal in anti-carbohydrate antibody production.

Feminization and reduction of testicular weight in mouse sparganosis

  • Yang, Hyun-Jong
    • Parasites, Hosts and Diseases
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    • v.44 no.2 s.138
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    • pp.167-169
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    • 2006
  • After infection of male mice with the plerocercoids (spargana) of Spirometra mansoni, serum levels of estrogen and testicular weight were analyzed by enzyme-linked immunosorbent assay (ELISA) and weighing machine, respectively. The serum level of estrogen increased progressively in infected mice compared with normal controls, whereas the testicular weight of infected mice decreased significantly (P < 0.05). These results suggest that certain substances from spargana change the steroid hormone metabolisms in the host by unknown pathways, and chronic infection may contribute to change of the function of steroid hormone target organ, i.e., testis, in male mice.

Separation of the Syncytial Layer of Spargana using Urea

  • Yang, Hyun-Jong
    • Parasites, Hosts and Diseases
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    • v.47 no.1
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    • pp.69-71
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    • 2009
  • The tegument of tapeworms is known to be composed of an outer syncytial cytoplasm layer which includes microtriches and cytoplasmic organelles (= syncytial layer), and a parenchymatous cytoplasm layer that contains subtegumental cell nuclei (= subtegumental layer) and organelles. In the present study, separation of the syncytial layer of the sparganum, the plerocercoid stage of Spirometra mansoni, was tried using urea as the chemical reagent. Histological sections were prepared to visualize the status of separation after staining with hematoxylin and eosin. The results showed that the syncytial layer of the sparganum tegument which includes microtriches and cytoplasmic organelles were successfully separated from the parenchyma using 3 M urea.

Production of Polyclonal Antibodies against the Tegument of Sparganum (Plerocercoid of Spirometra mansoni) and Its Immunolocalization

  • Yang, Hyun-Jong
    • Parasites, Hosts and Diseases
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    • v.48 no.2
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    • pp.183-185
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    • 2010
  • In a previous study, the author developed a method for separation of the tegument of spargana (plerocercoids of Spirometra mansoni) from the parenchyme using urea. The present study, as a next step, was performed to evaluate which molecules are present in the outer tegument. Two major proteins, 180 and 200 kDa, are present in the tegument and we could make polyclonal antibodies against these molecules. Their immunolocalization was processed and the outermost layer of the spargana showed strong positive staining. Conclusively, we could confirm that the 180 and 200 kDa molecules might be tightly bound membrane proteins in the tegument of spargana.

Immunoblot findings of calcareous corpuscles binding proteins in cyst fluid of Taenia solium metacestodes

  • Yang, Hyun-Jong
    • Parasites, Hosts and Diseases
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    • v.42 no.3
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    • pp.141-143
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    • 2004
  • After collecting calcareous corpuscles from plerocercoid of Spirometra mansoni (sparganum), we evaluated the antigenic values of calcareous corpuscles binding proteins obtained from the cyst fluid of Taenia solium metacestodes. Immunoblot analysis revealed that cysticercosis patient sera strongly recognized 10 and 95 kDa calcareous corpuscles binding proteins. This result demonstrated that calcareous corpuscles are bound with major secretory antigenic proteins, which is possibly involved in the secretory pathways of the 10 and 95 kDa proteins presenting in the cyst fluid of T. solium metacestodes.

Component Proteins and Protease Activities in Excretory-Secretory Product of Sparganum (스파르가눔 분비배설항원의 단백질 봉성 및 단백질분해효소 활성)

  • Cho, Seung-Yull;Chung, Young-Bae;Kong, Yoon
    • Parasites, Hosts and Diseases
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    • v.30 no.3
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    • pp.227-230
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    • 1992
  • Spirometra mansoni plerocercoid (sparganum) was incubated in saline at $4^{\circ}C{\;}or{\;}37^{\circ}C$ up to 100 hours. Protein contests in the excretory$.$secretory product (ESP) were rather constant (mean 7.7 mg of protein/gram of sparganum) in the preparations. Reducing SDS-PAGE of ESP showed similar protein subunit compositions with those in crude extract. Antigenic 36 and 31 kDa Proteins were major bands in ESP. ESP exhibited specific activities of protease(2.9~5.3 units/mg) at pH 6.0 and pH 7.5. Presence of protease activity in ESP may be a supporting evidence that hitherto known cysteiRe protease of sparganum is possibly secreted.

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Single step purification of potent antigenic protein from sparganum by gelatin-affinity chromatography (젤라틴 친화성 크로마토그래피를 이용한 스파르가눔 성분단백질의 순수분리)

  • Yoon Kong;Shin-Yong Kang;Seung-Yull Cho
    • Parasites, Hosts and Diseases
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    • v.29 no.1
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    • pp.1-8
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    • 1991
  • Out of many component proteins in crude saline extract of Spirometra mansoni plerocercoid (sparganum) , 36 kDa and 29 kDa proteins were found to be the most antigenic and were already purified by immunoaffinity chromatography using monoclonal antibody as a ligand. In this study, a single step purification of these potent antigenic proteins of sparganum extract was investigated. When the crude saline extract was charged to gelatin-Sepharose 4B affinity column, 36 kDa and 29 kDa protein fractions were bound. SDS-polyacrylamide gel electrophoresis (PAGE) and SDS-PAGE/immunoblot confirmed that the bound protein to gelatin was serologically pure. When evaluated by ELISA with patients sera, the purified protein of 36 and 29 kDa also showed improved antigenicity.

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IgG antibody responses in early experimental sparganosis and IgG subclass responses in human sparganosis

  • Chung, Young-Bae;Kong, Yoon;Yang, Hyun-Jong;Cho, Seung-Yull
    • Parasites, Hosts and Diseases
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    • v.38 no.3
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    • pp.145-150
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    • 2000
  • Antigenic components in the crude extracts of Spirometra mansoni plerocercoid were analyzed in early experimental infections and in IgG subclass observed in clinical sparganosis. By IgG immunoblot, sera obtained serially from experimental mice, fed 5 spargana each, were reacted with the crude extracts. Protein bands at 36-26 kDa and 103 kDa showed positive reactions since two weeks after infection. In a differential immunoblot, in which a monospecific antibody against sparganum chymase at 36 kDa was pre-treated, the reactions at 36-26 kDa disappeared, indicating that the sparganum chymase and its degradation products invoked IgG antibody reactions. When 69 patients sera of human sparganosis were examined for their IgG subclass responses, IgG4 levels showed the highest reaction which was followed by IgG 1 The IgG4 antibody also reacted mainly with 36-31 kDa protease. These results indicate that 36 kDa chymase of 5. nansoni plerocercoid is the main antigenic component inducing Ige antibody response in early stage of experimental sparganosis and for specific IgG subclass reactions in human sparganosis.

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