• Parasites, Hosts and Diseases
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• 제38권1호
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• pp.17-23
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• 2000

Eosinophils are important effector cells in host defense against parasites. Excretory-secretory product (ESP) produced by helminthic worms plays important roles in the uptake of nutrients, migration in the host tissue, and in immune modulation. However little is known about the ability of the ESP to directly trigger eosinophil apoptosis. This study investigated whether the ESP of newly excysted metacercariae of Paragonimus westermani could induce apoptosis in human eosinophils. Apoptosis was assayed by staining the cells with FITC-annexin V, and the cells were analyzed by flow cytometry. It was found that the ESP of newly excysted metacercariae of P. westemani induced a direct time- and concentration-dependent increase in the rate of constitutive apoptosis in mature human eosinophils. Eosinophil apoptosis was first apparent 3 hr after treatment with the ESP and continued to increase after 6 hr of incubation with respect to the cells cultured in the absence of the ESP. While only 2.8% of the eosinophils incubated in the medium for 3 hr were apoptotic, 7.6%, 10.9% and 22.6% of the eosinophils treated with 10. 30 and $100{\;}\mu\textrm{g}/ml$ ESP were apoptotic, respectively. This result suggests that the ESP of newly excysted metacercariae of P. westermani directly induce eosinophil apoptosis, which may be important for the survival of the parasites and the reduction of eosinophilic inflammation in vivo.

• Parasites, Hosts and Diseases
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• 제38권3호
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• pp.173-175
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• 2000

The present study examined postmetacercarial changes in the excysted metacercariae of Echinostoma caproni maintained in the defined medium Mixture 199 plus 20% calf serum for 7 days at $41^{\circ}C$. The gas phase was atmospheric air. Each culture was inoculated with 25 excysted metacerariae. Cultures were maintained upright in closed 15 ml plastic centrifuge tubes each containing 10 ml of medium plus 200 units of penicillin/ml and $200{\;}\mu\textrm{g}$ of streptomycin/ml. By 4 days in culture, most metacercariae had voided their excretory concretions. Organisms were clumped or solitary at the bottom of the cultures. Many organisms showed flaring of the oral collar and extension of both the collar and tegumentary spines. By 4 days in culture, posterior protuberances or bumps were noted on many of the organisms and some organisms showed abnormal vesicular growths or blebs at their posterior ends. Some mortality was noted in culture by day 5, but most organisms were still alive when the cultures were terminated on day 7.

• 한국어병학회지
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• 제13권2호
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• pp.137-142
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• 2000

간흡충 탈낭유충에 대한 잉어, 붕어, 참붕어의 보체 살충능 및 식세포 chemiluminescent 반응 차이를 조사하였다. 낙동강에서 채집한 참붕어는 모두 간흡충 피낭유충에 중감염되어 있었으나, 진영의 저수지에서 채집한 참붕어 및 낙동강에서 채집한 붕어와 양식 잉어에서는 간흡충 피낭유충이 전혀 검출되지 않았다. 각 실험어의 신선혈청을 이용하여 간흡충 탈낭유충에 대한 살충능을 조사한 결과 간흡충 피낭유충에 중감염된 참붕어 혈청과 붕어의 혈청은 잉어의 혈청에 비해 높은 살충능을 나타낸 반면, 간흡충 피낭유충에 감염되지 않은 참붕어의 혈청은 가장 낮은 살충능을 나타냈다. 각 어종의 혈청을 $47^{\circ}C$에서 30분간 처리하여 보체를 불활화시킨 결과 모든 실험어종에서 혈청의 살충능이 완전히 사라졌다. 간흡충 탈낭유충 균질액의 상층액을 각 어종의 두신 식세포에 첨가하여 chemiluminescent(CL) 반응을 측정한 결과 참붕어와 잉어에서 CL반응이 zymosan만을 첨가했을때에 비해 유의적(P<0.05)으로 감소하였으며, 각 어종별 간흡충 탈낭유충 균질액의 상충액 첨가에 의한 CL반응 감소율은 참붕어의 경우 22.9%, 붕어 9.6%,잉어 12.4%로 나타났다.

• Parasites, Hosts and Diseases
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• 제29권1호
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• pp.43-54
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• 1991

폐홉충(Paragonimus westermani) 초기 감염시의 숙주 면역 기전을 알아보기 위하여 폐흡충 피낭유충을 백서에 감염시켜 주별(주별)로 항혈청을 분리하고 등일 백서에서 복강 대식세포(homologous rat peritoneal macrophage)를 얻은 다음, 이들과 폐흡충 탈낭유충 및 숙주 조직 이행중인 유약충으로 effector system을 조작하여 폐흡충 유충에 대한 항체 의존성 대식세포 공격기전을 관찰하였다. 백서 복강 대식세포는 항혈청 출현 시에만 폐흡충 탈낭유충에 대한 세포부착 반응(cell adherence reaction)을 보였고 이 반응에는 보체가 관여하지 않았다. 폐흡충 감염 1주에서 8주까지의 백서 항혈청 중 2주째에 분리한 항쳔청 실험군에서 100%의 세포부착 반응을 나타내었고 사멸된 충체를 관찰할 수 있었으며, 세포부착 반응은 탈낭 유충기에서만 나타나는 반면 조직이행 유약충에서는 전혀 그 반응을 인정할 수 얼어 폐흡충 감염에 따른 항체의존성 숙주 세포 매개독성(antibodydependent cell·mediated cytotoxicity)은 발육단계 별 특이성을 보였다. 한편, 대식세포의 공격에 의하여 사멸된 폐흡충 탈낭유충의 형태학적 특징은 대식세포와 유충 사이에 형성되는 fuzzy material, tegumental syncytium 내의 미세구조의 변성 및 syncytium과 근육층을 연결하는 tubular tunnel의 형성 등이었다.

• Parasites, Hosts and Diseases
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• 제38권2호
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• pp.103-106
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• 2000

It is well known that the cysteine proteases in excretory-secretory product (ESP) of Paragonimus westermani newly excysted metacercariae (PwNEM) are capable of degrading IgG in vitro. Recent evidence suggests that the IgG-coated surface, such as found on parasites, is one of the most effective physiologic stimuli for granulocyte activation. Therefore, this study was designed to investigate the effect of excretory-secretory product (ESP) of PwNEM on superoxide production of granulocytes stimulated with IgG The 96-well plates were coated with human IgG (0, 10, 30, $100{\;}\mu\textrm{g}/ml$) in the absence or presence of ESP. When granulocytes were incubated in the wells coated with human IgG in the presence of ESP, the level of superoxide production of granulocytes was reduced to about 90% when compared to the cells incubated in the wells coated with IgG alone. This inhibitory effect of the ESP on IgG-induced superoxide production of granulocytes was concentration-dependent. These results suggest that ESP secreted by PwNEM may be important in the control of effector functions of granulocytes stimulated with IgG in human paragonimiasis.

• Parasites, Hosts and Diseases
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• 제45권3호
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• pp.199-204
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• 2007

Metacercariae of Gynaecotyla squatarolae (Digenea: Microphallidae) were discovered from the shore crab, Macrophthalmus dilatatus, purchased at a market in a coastal town of Taean-Eup, Chungcheongnam-do, Republic of Korea. Their adult flukes were confirmed by experimental infection of rats. The metacercariae of G. squatarolae were elliptical ($391.1{\times}362.5{\mu}m$), and the excysted metacercariae had progenetic genital organs, including the ovary and testes. To obtain adult flukes, 6 Sprague-Dawley rats were fed 500 metacercariae each, and killed at days 2, 4, and 6 post-infection. The adult flukes were identified as G. squatarolae (Yamaguti, 1934) Yamaguti, 1939, based on morphological characters, including 2 ventral suckers (1 large and 1 small), a large genital atrium equipped with the cirrus and the metraterm, separated male and female genital pores, a transversely long cirrus pouch, and extensive vitelline follicles. In the present study, it has been first proven that the shore crab M. dilatatus is a second intermediate host for G. squatarolae in the Republic of Korea.

• Parasites, Hosts and Diseases
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• 제36권4호
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• pp.217-225
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• 1998

미야타흡충 (Metagonimus miyatai) 피낭유충 및 성충의 표피 미세구조를 주자전자현미경으로 관찰하였다. 피낭유충은 피라미 (Zacco platypus)의 비늘에서 분리하였고, 성충은 흰쥐에 피낭유충을 감염시킨 후 1-4주에 소장으로부터 회수하였다. 탈낭한 피낭유충의 전체적인 외형은 요코가와흡충과 큰 차이가 없었으며, 복흡반은 충체 중앙으로부터 오른쪽에 위치하였다. 구흡반의 구순에는 제I형 감각유두 및 제II형 감각유두가 여러 개 관찰되었다. 복흡반 앞쪽에는 끝이 5-7분지된 피극들이 빽빽하게 덮혀 있었으며, 충체 후반부의 배설공 부근에는 2-3분지된 피극으로 덮혀 있었다. 감염 1주된 성충의 경우, 구흡반의 구순에는 제II형 감각유두 7개가 분포하고 있었으며, 구흡반 내벽에는 큰 제I형 감각유두가 1쌍씩, 작은 제I형 감각유두가 2쌍씩 좌우측에서 각각 관찰되었다. 피극의 분포는 피낭유충의 경우와 비슷하였으나, 분화도는 증가하여 9-11분지된 피극들이 관찰되었다. 감염 2-4주된 성충의 표피 미세구조는 감염 1주된 성충과 대동소이하였으나, 정자가 충체 배측 표면으로 개구된 Laurer＇s canal로 들어가고 있는 것이 관찰되었다. 이상의 결과를 종합할 때, 미야타흡충의 표피 미세구조는 요코가와흡충과 전반적으로 비슷하였으나 피극의 분포 및 분화도, 구흡반 주변의 감각유두 분포 등에 다소 차이가 있음을 알 수 있었으며 이러한 차이가 분류학적으로 의미있는 것일 가능성이 제시되었다.

• Parasites, Hosts and Diseases
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• 제34권4호
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• pp.283-285
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• 1996

196년 9월 7일 전남 목표항에서 구입한 붕장어의 아가미 새엽에서 6개체의 피낭유충이 검출되었다. 이중 탈낭된 한 개의 형태를 관찰한 바 긴이형흡충의 피낭유충으로 동정되었으며, 붕장어가 긴이형흡층의 제2중간숙주 역활을 한다는 것으로 보고있다.

• 한국임상수의학회지
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• 제25권4호
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• pp.307-309
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• 2008

The metacercariae species have often been isolated from fresh water fish in Korea. In this study, we reported Clinostomum complanatum, a species of metacercariae isolated from gill and muscle of Korean barbel (Hemibarbus mylodon), a natural monument fresh water fish in Korea, by morphological observation. The excysted metacercariae, tongue-shaped and progenetic, were 3.11-4.62 mm in length and 0.90-1.43 mm in width. Fresh water fishes are known as the second intermediate hosts of C. complanatum. This is the first isolation report from Korean barbel.

• Parasites, Hosts and Diseases
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• 제46권2호
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• pp.95-99
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• 2008

Eosinophil degranulation plays a crucial role in tissue inflammatory reactions associated with helminth parasitic infections and allergic diseases. Paragonimus westermani, a lung fluke causing human paragonimiasis, secretes a large amount of cysteine proteases, which are involved in nutrient uptake, tissue invasion, and modulation of hos's immune responses. There is, however, limited information about the response of eosinophils to direct stimulation by cysteine proteases (CP) secreted by P. westermani. In the present study, we tested whether degranulation and superoxide production from human eosinophils can be induced by stimulation of the 2 CP (27 kDa and 28 kDa) purified from excretory-secretory products (ESP) of P. westermani newly excysted metacercariae (PwNEM). A large quantity of eosinophil-derived neurotoxin (EDN) was detected in the culture supernatant when human eosinophils isolated from the peripheral blood were incubated with the purified 27 kDa CP. Furthermore, the 27 kDa CP induced superoxide anion production by eosinophils in time- and dose-dependent manners. In contrast, the purified 28 kDa CP did not induce superoxide production and degranulation. These findings suggest that the 27 kDa CP secreted by PwNEM induces superoxide production and degranulation of human eosinophils, which may be involved in eosinophil-mediated tissue inflammatory responses during the larval migration in human paragonimiasis.