• Title/Summary/Keyword: Excretory-secretory

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Degradation of human immunoglobulins and cytotoxicity on HeLa cells by live Trichomonas vaginalis (질편모충 단백질분해효소의 세포독성 및 인체면역글로불린 분해능)

  • 민득영;류재숙
    • Parasites, Hosts and Diseases
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    • v.35 no.1
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    • pp.39-46
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    • 1997
  • The present study was undertaken to determine whether live T. uaginnlis degrades human secretory IgA, serum If and IgG molecules. Human immunoglobulins were exposed to live trophozoites, parasite Iysate, and excretory-secretory product (ESP) of T ucginnlis. To determine the fragmentation of immunoglobulins, the reaction sample was subjected to SDS-PAGE and EITB, and peroxidase conjugated antihuman IgA and IgG were used as probes. Live trophozoites degraded secretory IgA, serum IgA and IgG, and degradation were pressed forward by the prolongation of the incubation time and by increasing the number of trichomonads respectively. Also the Iysates and ESP of trichomonads degraded IgA and IgG. The cysteine and serine proteinase inhibitors such as I-64, antipain, iodoacetic acid, iodoacetamide, TLCK reduced the ability of cleaving immunoglobulins. The proteinase activity and cytotoxicity of T. uaginnlis to HeLa cells were decreased when live T. vusinalis was treated with metallo-proteinase inhibitor as well as cysteine and serine proteinase inhibitors. These results suggest that proteinase secreted from live T ucginclis may play a part role in host pathogenesis by T. uosinnlis, and the cleaving ability of host immunoglobulins by the proteinase may contribute as a one of immune evasion mechanism for parasite survival in the host.

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Immunohistoehemical Observation on the Antigens Inducing IgG and IgM Antibodies against Sparganum (IgG와 IgM 항체를 유도하는 sparganum의 항원에 관한 면역조직화학적 및 전기영동에 의한 연구)

  • 김창환;최완성
    • Parasites, Hosts and Diseases
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    • v.29 no.4
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    • pp.339-354
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    • 1991
  • Localization and characterization of the antigenic components of sparganum which induced IgG and IsM antibodies in the host were studied by immunohistochemical techniques and SDS-PAGT and Western blotting. The antigen recognized by IgG antibody of rats or mice which were immunised by infection or injection of crude extracts of metacestodes of Spirometra erinacei, was located in the parenchyme of sparganum, especially at the cortex and around the calcareous corpuscles. The immunoreaction was demonstrated not only in the encysted fibrous wall of host but around the arterioles or venules in the connective tissue of host. The antigen recognized by IgM antibody of rats or mice was also observed in the parenchyme of sparganum and in the connective tissue of host. By 5∼20% gradient SDS-PAGE and EIBT, we detected antigenic components by IgG and 1gG antibodies of the rat or mouse immunized by infection or injection of crude extract of spargana. Twenty-three antigenic bands from crude extracts of spargana were recognized by IgG antibody and 15 components by IgM antibody of immunized rats. Out of the bands recognized by IgG and IgM antibodies, 15 were cross-reacted each other. Twenty components of eBlcretory-secretory proteins from spargana were recognized by IgG, and 5 components by IgM antibody of immunized rats. By IgG and IgM antibodies of immunized mice, 16 components of crude extracts were recognized by IgG antibody and 9 components by IgM antibody. Twenty components of excretory-secretory preparation were recognized by IgG antibody and 5 components by IgM antibody. Thirteen components of crude extracts were cross-reacted by IgG antibody of rats and mice.

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Postnatal development of Harderian gland in Mongolian gerbil (몽고리안 저빌의 Harderian gland의 출생후 형태학적 변화)

  • Oh, Seung-hyun;Park, Ji-young;Yoon, Yeo-sung;Kim, Dae-joong;Nam, Sang-yoon;Lee, Joon-sup;Seong, Je-kyung
    • Korean Journal of Veterinary Research
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    • v.39 no.1
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    • pp.1-11
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    • 1999
  • This investigation was carried out to study morphological and chronological aspects of the development of the Harderian gland in the Mongolian gerbil(Meriones unguiculatus). Male and female Mongolian gerbils were sacrificed on days 1, 3, 5, 10, 30 and 60 after birth and their Harderian glands were processed for light microscopic observation. The results obtained were summarized as follows; 1. In 1-day-old Mongolian gerbil, Harderian gland was well distinguished from other tissue structures. It was composed of several immature tubules, and these tubules were separated each other by undifferentiated mesenchymal connective tissues. 2. In 3-day and 5-day-old Mongolian gerbils, the arrangement of tubules in the gland was more condensed than that of 1-day-old Mongolian gerbil. The excretory ducts started to appear in the connective tissues located between lobes. 3. In 10-day-old Mongolian gerbil, small lipid vacuoles began to be found in the cytoplasm of the secretory cells of the Harderian gland. There were some mucus-secreting cells within the epithelium of the excretory duct found in the interlobar connective tissues. 4. In 30-day-old Mongolian gerbil, there was markedly increased number of the tubules in the glands. The epithelial cells of the tubules were typically columnar in shape. Most of the columnar epithelial cells contained many small lipid vacuoles, although a few cells contained large lipid vacuoles. 5. In 60-day-old Mongolian gerbil, the Harderian gland exhibited the typical structural characteristics of the adult gland. The mature glandular structures were more distinct than those of 30-day-old animals.

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Ultrastructural studies on the surface of Puragonimus westermani metacercaria (폐흡충 피낭유충 표피의 형태학적 관찰)

  • Kim, Gyeong-Min;An, Myeong-Hui;Min, Deuk-Yeong
    • Parasites, Hosts and Diseases
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    • v.25 no.2
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    • pp.129-140
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    • 1987
  • The present study was undertaken to demonstrate the surface structure of Paragonimus westermani metacercaria in Korea with special reference to the distribution of sensory papillae. Metacercariae were isolated from crayfish, one of the second intermediate tost of P. westermani in Bogil island, Chollanam-do (Province), Korea, where has been known as an endemic area of human paragonimiasis. Isolated metacercariae were encysted and examined with light. scanning and transmission electron microscopes for morphological features. On the surface of iBetacercariae, three types of sensory FaFillae were identified. Large domed papillae ($3~5{\mu\textrm{m}}$), which were covered with wrinkled plasma n!embrane of the worm, were distributed on the oral and ventral suckers only. On the oral sucker, these large domed papillae were 12~13 in number. On the other hand large domed papillae on the ventral sucker were constantly 6 in number and hexagonal in distribution. Small domed papillae ($2~3{\mu\textrm{m}}$), of which surface was more smooth than those of large ones, were distributed symmetrically on the ventral (30~32 pairs) and dorsal surfaces (40~42 Pairs). Ciliated Papillae ($0.8~1.5{\mu\textrm{m}}$) were observed about 5~6 in number around the oral sucker and 3~5 pairs each on the ventral and dorsal surface of the body. Single Fcinted spines covered the entire surface of the body except around the excretory pore. Spines on the anterior fart of the body were 0.9~2.0${\mu\textrm{m}}$ in length and $45~55/100{\mu\textrm{m}}$2 in number, and were gradually reduced in length ($0.4~1.4{\mu\textrm{m}}$) and in nuns.her ($12~27/100{\mu\textrm{m}}$2) toward the posterior part. The body wall of p. westermoni metacercariae was consisted with anucleated syncytium layer, fibrous interstitial layer and musclar layer. In the anucleated syncytium, biconcave ($0.15~0.55{\mu\textrm{m}}$) and spherical ($0.08~0.16{\mu\textrm{m}}$) secretory granules, which were transferred from epidermal cells via protoplasmic tubules, mitochondria and rihoEorses, T-ere observed. Spines originated around the basement membrane protruded externally. Epidermal cells were consisted with a nucleus and a cytoplasm, and connected to syncytium with protoplasmic tubules. In the cytoplasm many secretory granules, mitochondria, Golgi complex, endoplasmic reticula, ribosomes and lipid droplets were observed.

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Organ-specific antigens of Clonorchis sinensis

  • Li, Shun-Yu;Chung, Byung-Suk;Choi, Min-Ho;Hong, Sung-Tae
    • Parasites, Hosts and Diseases
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    • v.42 no.4
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    • pp.169-174
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    • 2004
  • This study was carried out to find out specific proteins from different organs of Clonorchis sinensis. Crude extract, organ-specific and excretory-secretory (ES) proteins were analyzed by immunoblot with infected human sera. The bands of 7- and 17 -kDa were main component of intestinal fluid and ES protein and commonly found in all organ-specific proteins. The 17-kDa protein was observed from ES antigen, intestinal fluid, eggs and sperms, 26- and 28-kDa proteins were from the uterus, vitellaria, and ovary, and 34-, 37-, 43- and 50-kDa proteins were mainly from the testis and sperms. Serum of mice immunized with sperms reacted to the 50-kDa protein by immunoblotting and immunohistochemical staining showed a positive reaction at the seminal receptacle and seminiferous tubule. The present results show that the 7-kDa protein is a common antigen of every part or organ of C. sinensis, but different organs express their specific antigenic protein bands.

Expression on Echinostoma hortense Infection Manifestation on the Cytokine of the Splenocytes of BALB/c and C3H/HeN Mice

  • Ryang Yong-Suk;Cho Yoon-Kyung;IM Jee-Aee;Lee Kyu-Je;Kim Insik;Sung Ho-Joong
    • Biomedical Science Letters
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    • v.10 no.3
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    • pp.219-230
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    • 2004
  • This experiment was performed to examine the in vitro and in vivo affects of the two different haplotype strains of mice, BALB/c and C3H/HeN infected with Echinostoma hortense, and the manifestation of the profiles of cytokine in the splenocytes. In the in vitro experiment, the two mice's splenocytes were divided and stimulated with antigen of crude extracts and the antigen of excretory and secretory products of an adult warm and the manifestation of cytokine mRNA was verified with RT-PCR. As a result, the two different strains of mice both strongly manifested the Th2 cytokine rather than the Thl cytokine and in the case of the Th2 cytokine, the BALB/c mice manifested more strongly than the C3H/HeN mice. In the experiment using the ELISA method, the protem cytokine manifestation had the same result as the mRNA experiment. In the in vivo experiment, the mice was infected via oral route with the metacercaria of the Echinostoma hortense and the manifestation of cytokine was verified by RT-PCR and ELISA and the results were the same as the in vitro experiment. Therefore, in the two strains of BALB/c and C3H/HeN, the C3H/HeN showed a higher susceptivity to the Echinostoma hortense.

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Serodiagnosis of Toxocariasis by ELISA Using Crude Antigen of Toxocara canis Larvae

  • Jin, Yan;Shen, Chenghua;Huh, Sun;Sohn, Woon-Mok;Choi, Min-Ho;Hong, Sung-Tae
    • Parasites, Hosts and Diseases
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    • v.51 no.4
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    • pp.433-440
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    • 2013
  • Toxocariasis is a worldwide zoonosis caused by larvae of ascarid nematodes of dogs or cats, Toxocara canis or T. cati. Diagnosis of human toxocariasis currently relies on serology that uses T. canis excretory-secretory antigen to detect specific IgG antibodies by ELISA. We investigated the serodiagnostic efficacy of ELISA using crude antigen of T. canis larvae (TCLA). Serum specimens of 64 clinically confirmed toxocariasis, 115 healthy controls, and 119 other tissue-invading helminthiases were screened by ELISA using TCLA. The ELISA using TCLA showed 92.2% (59/64 patient samples) sensitivity and 86.6% (103/119) specificity. Its positive diagnostic predictivity was 78.7% and negative predictivity was 97.8%. No serum of healthy controls reacted but that of anisakiasis (45.5%), gnathostomiasis (19.2%), clonorchiasis (15.8%), sparganosis (11.1%), and cysticercosis (6.3%) cross-reacted. Immunoblot analysis on TCLA recognized antigenic proteins of 28- and 30-kDa bands in their dominant protein quantity and strong blotting reactivity. The present results indicate that the ELISA using our TCLA antigen is acceptable by the sensitivity and specificity for serodiagnosis of human toxocariasis. ELISA with TCLA is recommended to make differential diagnosis for patients with any sign of organ infiltration and eosinophilia.

Comparative Characterization of Four Calcium-Binding EF Hand Proteins from Opisthorchis viverrini

  • Emmanoch, Palida;Kosa, Nanthawat;Vichasri-Grams, Suksiri;Tesana, Smarn;Grams, Rudi;Geadkaew-Krenc, Amornrat
    • Parasites, Hosts and Diseases
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    • v.56 no.1
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    • pp.81-86
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    • 2018
  • Four isoforms of calcium binding proteins containing 2 EF hand motifs and a dynein light chain-like domain in the human liver fluke Opisthorchis viverrini, namely OvCaBP1, 2, 3, and 4, were characterized. They had molecular weights of 22.7, 21.6, 23.7, and 22.5 kDa, respectively and showed 37.2-42.1% sequence identity to CaBP22.8 of O. viverrini. All were detected in 2- and 4-week-old immature and mature parasites. Additionally, OvCaBP4 was found in newly excysted juveniles. Polyclonal antibodies against each isoform were generated to detect the native proteins in parasite extracts by Western blot analysis. All OvCaBPs were detected in soluble and insoluble crude worm extracts and in the excretory-secretory product, at approximate sizes of 21-23 kDa. The ion-binding properties of the proteins were analyzed by mobility shift assays with the divalent cations $Ca^{2+}$, $Mg^{2+}$, $Zn^{2+}$, and $Cu^{2+}$. All OvCaBPs showed mobility shifts with $Ca^{2+}$ and $Zn^{2+}$. OvCaBP1 showed also positive results with $Mg^{2+}$ and $Cu^{2+}$. As tegumental proteins, OvCaBP1, 2, and 3 are interesting drug targets for the treatment of opisthorchiasis.

The histological structure and the pathological lesions of gill in teleosts (어류의 아기미의 조직학적 구조와 병변)

  • Huh, Min-Do;Jeong, Hyun-Do
    • Journal of fish pathology
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    • v.6 no.1
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    • pp.65-70
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    • 1993
  • The delicate histological structure of gill in teleosts can be easily affected by a variety of biological, chemical or physical detrimental agents because it is directly exposed to the surrounding water. The epithelium of secondary lamella is thin to allow efficient gaseous exchange and this also renders it particularly vulnerable to various pathogens. As well as the main respiratory role, the gill has other various important functions such as acid-base balance, osmoregulation or the excretion of nitrogenous waste products. Thus destruction of epithelial integrity such as epithelial necrosis or thickening can render a fish very vulnerable to respiratory, secretory and excretory difficulties. This article was tried to describe in detail the common processes of pathological responses correlated to the normal histological structures of the gill in teleosts.

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A 27 kDa Cysteine Protease Secreted by Newly Excysted Paragonimus westermani Metacercariae Induces Superoxide Anion Production and Degranulation of Human Eosinophils

  • Chung, Young-Bae;Kita, Hirohito;Shin, Myeong-Heon
    • Parasites, Hosts and Diseases
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    • v.46 no.2
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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.