• 대한수의학회지
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• 제39권1호
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• pp.159-168
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• 1999

Swine whipworm(Trichuris suis) is cosmopolitan nematode which can cause serious pathology in immature stage(larva2~larva5) of infected pigs, such as anorexia, diarrhea, anemia, and death in heavy infections. In this larval stages, it is very difficult to diagnose the infection of whipworm and to differentiate from other common swine gastrointestinal disorders such as 21 day scours which are associated with TGE virus, rota virus, coccidium, and the stress of weaning. In this experiment, the isolated antigens of Trichuris spp. were carried out to examine the structure and specificity of antigens and to select the reasonable antigens which would be used in serological diagnosis by electrophoresis, Western blotting, ELISA. The results of this experiment were as follows; 1. The common fractions of each Trichuris suis antigen were identified 28,32,45, 80kDa by SDS-PAGE with silver stain and four major fractions could be detected in positive swine sera by Western blot analysis. 2. The OD(optical density) values of somatic and excretory-secretory antigens which were reacted against positive(negative) sera from pigs infected with Trichuris suis by ELISA reader were; 1) OD values($mean{\pm}SD$) of adult somatic antigen against positive(negative) sera were $0.30{\pm}0.12(0.09{\pm}0.006)$ and third-stage larva of somatic antigen were $0.28{\pm}0.038(0.10{\pm}0.005)$. And OD values of excretory-secretory antigens of adult and third-stage larva were $0.24{\pm}0.031(0.11{\pm}0.005)$ and $0.08{\pm}0.013(0.10{\pm}0.003)$, respectively. 2) OD values of adult somatic, larval somatic antigen and adult excretory-secretory antigen response to positive sera were significantly (p<0.01) associated with negative swine sera. And the Cut-off OD values(minimum positive value) were determined to be mean negative value plus 3 SD that would minimized the risk of false positives. 3. The OD values of somatic antigens of T suis and T vulpis against swine positive(negative) sera were $0.30{\pm}0.120(0.09{\pm}0.006)$ and $0.25{\pm}0.141(0.09{\pm}0.003)$. These data mean that the somatic antigens of T suis and T vulpis were able to diagnose T vulpis infection in dogs as well as T suis infection in pigs. These results suggest that somatic antigen of third-stage larva and excretory-secretory antigen of adult T suis could be used the diagnostic antigen by serological test(ELISA) in immature Trichuris spp. infection.

• Applied Microscopy
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• 제18권2호
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• pp.91-101
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• 1988

The ultrastructure of the sac and tail portion of the large ampullate gland and production of the silk materials in the orb web spider, Nephila clavata L. Koch, are studied with electron microscope. Large ampullate glands, the largest glands among the seven kinds of silk glands in this species, are composed of three parts which are the excretory duct, the storage sac and the convoluted tail. The wall of the sac is composed of a single layer of columnar epithelial cells. In the cytoplasm of these cells several kinds of the secretory granules, which are commonly associated with the rough endoplasmic reticula and had characteristic crystalloid, are seen. According to the morphology and internal textures of these granules, the sac portion is subdivided into proximal(connected with the convoluted tail) and distal(connected with excretory duct) portion. Between these two portions, the proximal portion is longer than the distal by four times. Silk materials, being synthesized to the shape of secretory granules, within the glandular epithelial cell of the tail portion are released to the inner cavity by the mechanism of the eccrine secretion. These secretory granules are originated from the rough endoplasmic reticula of the glandular epithelial cells, whereas no Golgi complexes has been found in any of the cells which have been examined.

• 한국동물학회:학술대회논문집
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• 한국동물학회 1995년도 한국생물과학협회 학술발표대회
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• pp.288.2-288
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• 1995

No Abstract, See Full Text

• Parasites, Hosts and Diseases
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• 제45권1호
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• pp.19-26
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• 2007

This study describes the isolation of a Toxocara canis species-specific excretory-secretory(ES) antigen and the development of an enzyme-linked immunosorbent assay(ELISA) based on this antigen. Analysis of the ES antigens of T. canis, Toxocara vitulorum, Ascaris lumbricoides and Necator americanus larval antigen was performed by SDS-PAGE followed by western blotting. A 57 kDa T. canis-specific antibody fraction(TcES-57) was identified by western blotting and labelling with anti-Toxocara antibodies(from experimental rabbits and human patients) and tracing with anti-human or anti-rabbit peroxidase conjugate. No protein fraction of 57 kDa was detected in ES or larval antigens collected from T. canis, T. vitulorum, A. lumbricoides and N. americanus. Using TcES-57, a specific anti-serum was produced in rabbits and a double sandwich ELISA was developed. This test was validated using known seropositive sera from toxocariasis patients, sera from A. lumbricoides or N. americanus patients, and 50 serum samples from cats. These tests revealed that TcES-57 antigen is specific to T. canis infection and does not cross react with sera of other related infections. Thus, ELISA based on TcES-57 antigen was proven to be an effective tool in the diagnosis of toxocariasis and studies on the role of T. canis in the epidemiology of human toxocariasis.

• Parasites, Hosts and Diseases
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• 제50권2호
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• pp.165-171
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• 2012

Larval excretory-secretory products of Anisakis simplex are known to cause allergic reactions in humans. A cDNA library of A. simplex 3rd-stage larvae (L3) was immunoscreened with polyclonal rabbit serum raised against A. simplex L3 excretory-secretory products to identify an antigen that elicits the immune response. One cDNA clone, designated as ${\alpha}$-methylacyl CoA racemase (Amacr) contained a 1,412 bp cDNA transcript with a single open reading frame that encoded 418 amino acids. A. simplex Amacr showed a high degree of homology compared to Amacr orthologs from other species. Amacr mRNA was highly and constitutively expressed regardless of temperature (10-$40^{\circ}C$) and time (24-48 hr). Immunohistochemical analysis revealed that Amacr was expressed mainly in the ventriculus of A. simplex larvae. The Amacr protein produced in large quantities from the ventriculus is probably responsible for many functions in the development and growth of A. simplex larvae.

• Parasites, Hosts and Diseases
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• 제46권3호
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• pp.127-132
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• 2008

Clonorchis sinensis is one of the most prevalent parasitic helminths in Korea. Although cholangiocarcinoma can be induced by C. sinensis infection, the underlying mechanism is not clearly understood. To assess the role of C. sinensis infection in carcinogenesis, an in vitro system was established using the human epithelial cell line HEK293T. In cells exposed to the excretory/secretory products (ESP) of C. sinensis and the carcinogen dimethylnitrosamine (DMN), cellular proliferation and the proportion of cells in the G2/M phase increased. Moreover, the expression of the cell cycle proteins E2F1, p-pRb, and cyclin B was dramatically increased when ESP and DMN were added together. Similarly, the transcription factor E2F1 showed its highest level of activity when ESP and DMN were added simultaneously. These findings indicate that DMN and ESP synergistically affect the regulation of cell cycle-related proteins. Our results suggest that exposure to C. sinensis and a small amount of a carcinogen such as DMN can promote carcinogenesis in the bile duct epithelium via uncontrolled cellular proliferation and the upregulation of cell cycle-related proteins.

• Parasites, Hosts and Diseases
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• 제57권4호
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• pp.379-387
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• 2019

Clonorchis sinensis is a carcinogenic human liver fluke that promotes hepatic inflammatory environments via direct contact or through their excretory-secretory products (ESPs), subsequently leading to cholangitis, periductal fibrosis, liver cirrhosis, and even cholangiocarcinoma (CCA). This study was conducted to examine the host inflammatory responses to C. sinensis ESPs and their putative protein components selected from C. sinensis expressed sequenced tag (EST) pool databases, including $TGF-{\beta}$ receptor interacting protein 1(CsTRIP1), legumain (CsLeg), and growth factor binding protein 2 (CsGrb2). Treatment of CCA cells (HuCCT1) with the ESPs or bacterial recombinant C. sinensis proteins differentially promoted the secretion of proinflammatory cytokines ($IL-1{\beta}$, IL-6, and $TNF-{\alpha}$) as well as anti-inflammatory cytokines (IL-10, $TGF-{\beta}1$, and $TGF-{\beta}2$) in a time-dependent manner. In particular, recombinant C. sinensis protein treatment resulted in increase (at maximum) of ~7-fold in $TGF-{\beta}1$, ~30-fold in $TGF-{\beta}2$, and ~3-fold in $TNF-{\alpha}$ compared with the increase produced by ESPs, indicating that CsTrip1, CsLeg, and CsGrb2 function as strong inducers for secretion of these cytokines in host cells. These results suggest that C. sinensis ESPs contribute to the immunopathological response in host cells, leading to clonorchiasis-associated hepatobiliary abnormalities of greater severity.

• 한국어병학회지
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• 제27권1호
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• pp.25-33
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• 2014

Anisakis simplex sensu stricto와 A. pegreffii의 3기 유충과 4기 유충에서 얻은 excretory-secretory (ES) products 및 somatic extracts의 가수분해효소 활성을 API ZYM kit를 이용하여 비교하였다. Esterase 그룹의 가수분해효소 중 acid phosphatase는 A. simplex (s.s.)와 A. pegreffii 모두에서 높은 활성을 나타냈으며, esterase (C 4)의 경우 somatic extracts에서만 가수분해효소 활성이 나타났는데 A. simplex (s.s.)가 A. pegreffii와 비교하여 3기와 4기 유충 모두에서 2배 가량 높은 활성을 나타냈다. alkaline phosphatase, acid phosphatase 그리고 naphthol-AS-BI-phosphohydrolase의 경우 A. simplex (s.s.)와 A. pegreffii 모두 3기 유충보다 4기 유충에서 더 높은 가수분해효소 활성이 확인되었다. Aminopeptidase 그룹의 가수분해효소 활성은 leucine arylamidase에서 관찰되었는데, somatic extracts의 경우 A. pegreffii 보다 A. simplex (s.s.)에서 가수분해 효소 활성이 두 배 가량 높게 확인되었으며, 대부분의 다른 효소들에서는 활성이 거의 나타나지 않았다. Glycosidase 그룹의 가수분해효소 활성은 N-acetyl-${\beta}$-glucosaminidase, ${\alpha}$-mannosidase 그리고 ${\alpha}$-fucosidase에서 확인되었는데, A. simplex (s.s.) 보다 A. pegreffii에서 높은 가수분해효소 활성을 확인할 수 있었으며, 대부분 4기 유충보다 3기 유충에서 더 높은 가수분해효소 활성이 확인되었다. 이러한 아니사키스속 선충의 종과 유충단계에 따른 가수분해 효소의 활성 차이는 선충의 성장, 탈피, 소화, 섭이 등의 대사과정의 차이에 기인한 것으로 생각된다.

• 대한의생명과학회지
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• 제3권2호
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• pp.89-94
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• 1997

가토에게 간흡충을 감염시키고 3개월 후 거살한 다음 간흡충의 충체, 간흡충의 분비배설액 그리고 간토의 담즙을 조항원으로 제조한 후 이에 대한 항원단백질의 구성물질을 분석하였다. 그리고 cysteine계 면역억제제인 E-64와 serine계 면역억제제인 PMSF를 첨가하였을 때 단백질 구성물질의 발현 양상을 관찰하였다. 실험적으로 가토에서 얻은 간흡충 성충의 조항원은 200-9 kDa의 범위에서 26개의 분획 을 관찰하였으며, 간흡충 성충 조항원에 cysteine계 단백질분해효소 억제제인 E-64를 첨거하였을 때 잘 보존하여 200-9 kDa의 범위에서 29개의 분획을 관찰하였다. 간흡충 성충의 분비배설액 조항원은 200-9 kDa범위에서 27개의 분획이 관찰되었으며, cysteine계 단백질분해효소 억제제인 E-64를 첨거하였을 때 잘 보존하여 200-9 kDa의 범위에서 29개의 분획이 관찰되었다. 그리고 간흡충을 감염시킨 가토의 담즙 조항원은 200-9 kDa의 범위에서 19개의 분획이 관찰되었으며, serine 계 단백질분해효소인 PMSF를 첨거하였을 때 200-10 kDa의 범위에서 22개의 분획이 관찰되었으나, cysteine계 단백질분해효소인 E-64에서도 비슷한 양상을 보였다. 대조군인 건강 가토의 담즙 조항원은 200-10 kDa의 범위에서 22개의 분획이 관찰되었으며, serine계 단백질분해효소 억제제인 PMSF를 첨가하였을 때 잘 보존하여 200-12 kDa의 범위에서 23개의 분획이 관찰되었다. 앞으로 각 항원에 대한 면역학적 특징 및 단세포군 항체에 대한 구체적인 규명이 계속되어야겠다.

• Applied Microscopy
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• 제19권1호
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• pp.49-58
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• 1989

Ultrastructure of the excretory duct of the small ampullate gland in the orb web spider, Nephila clavata L. Koch are studied with light and electron microscopes. The small ampullate glands, located near the midline portion of the abdominal cavity, are connected with the spigots(large spinning tubes) on the middle spinnerets and composed of three parts which are the excretory duct, the storage sac and the convoluted tail. The excretory duct of this gland is enclosed by a thin layer of the outer connective tissues. By the morphology of the apical cuticles and internal textures of the epithelial cells, the duct is subdivided into two regions which are proximal duct region near the sac and distal duct region near the spinnerets. At the distal region of the ducts, the subcuticle which had the function of water removal form the progenetive silk material is well developed, whereas at the proximal region this cuticle disappeared and instead of these, endocuticle is developed. Moreover the epithelium of the distal duct region is composed of columnar epithelial cells, but at the proximal region the epithelium is changed to squamous or cuboidal forms. In the cytoplasm of the epithelial cells, rough endoplasmic reticula, Golgi comlexes and large secretory vesicles related to the production of the cuticular materials are well developed. And between the adjacent epithelial cells, specialized septate junction and desmosomes are formed along the plasma membrane.