1.Functional Genes and Proteins of Clonorchis sinensis.
Tae Im KIM ; Byoung Kuk NA ; Sung Jong HONG
The Korean Journal of Parasitology 2009;47(Suppl):S59-S68
During the past several decades, researches on parasite genetics have progressed from biochemical and serodiagnostic studies to protein chemistry, molecular biology, and functional gene studies. Nowadays, bioinformatics, genomics, and proteomics approaches are being applied by Korean parasitology researchers. As for Clonorchis sinensis, investigations have been carried out to identify its functional genes using forward and reverse genetic approaches and to characterize the biochemical and biological properties of its gene products. The authors review the proteins of cloned genes, which include antigenic proteins, physiologic and metabolic enzymes, and the gene expression profile of Clonorchis sinensis.
Animals
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Clonorchiasis/parasitology
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Clonorchis sinensis/enzymology/*genetics/*metabolism
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Gene Expression Regulation
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Helminth Proteins/*genetics/*metabolism
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Humans
2.Metabolism of C(14)-glycine by Clonorchis sinensis.
Soo Hyun SEONG ; Byong Seol SEO
The Korean Journal of Parasitology 1966;4(2):14-22
Radioactive C(14)-glycine was given to Clonorchis sinensis in Tyrode medium in order to trace the metabolic fate of the labeled carbon. The labeled carbon from glycine enters into every major fraction of Clonorchis sinensis and is highest in the fraction of protein and nucleic acid. Significant amount of C(14)-glycine is incorporated into respiratory carbon dioxide. Relatively high percentage of C(14)-glycine in medium is converted to amino acid fraction and lipid fraction of the worm. In general, glycine is continuously being utilized in the synthesis of proteins and for energy production despite the uptake rate of glycine decreased gradually as incubation proceeds.
parasitology-helminth-trematoda
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Clonorchis sinensis
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metabolism
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biochemistry
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glycine
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amino acid
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nucleic acid
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protein
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lipid
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Tyrode medium
3.Study on the (14)C-glucose metabolism by Clonorchis sinensis.
Il Kwon KANG ; Soon Hyung LEE ; Byong Seol SEO
The Korean Journal of Parasitology 1969;7(3):143-152
Radioactive (14)C-glucose(U) was given to Clonorchis sinensis in Tris buffer medium, in corder to trace the metabolic fate of the labelled carbon. The labelled carbon from glucose enters into intermediary metabolites and end products of anaeroblic glycolysis, Embden-Meyerhof pathway, and of aerobic Krebs cycle. These product were identified by one or two-dimensional paper chromatography in combination with autoradoigraphy. The labelled metabolites detected in this experiment corresponded to pyruvic acid, latic acid, malic acid, succinic acid and fumaric acid. Amino acids, such as alanine, aspartic acid, glutamic acid, valine, theronine, and serine, derived by the degradation of (14)C- glycose were also found. Labelled compounds behaving like alanine, aspartic acid and glutamic acid were observed in the chroma to gram of incubation medium. The preciptation which suggests a positive reaction for protein occured when absolute ethanol was added to the incubation medium.
parasitology-helminth-trematoda
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Clonorchis sinensis
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two-dimensional paper chromatography
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autoradiography
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metabolism
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glucose
4.Ultrastructural localization of 28 kDa glutathione S-transferase in adult Clonorchis sinensis.
Sung Jong HONG ; Jae Ran YU ; Shin Yong KANG
The Korean Journal of Parasitology 2002;40(4):173-176
Glutathione S-transferase (28GST) with molecular mass of 28 kDa is an antioxidant enzyme abundant in Clonorchis sinensis. In adult C. sinensis, 28GST was localized in tegumental syncytium, cytons, parenchyma, and sperm tails examined by immunoelectron microscopy. C. sinensis 28GST was earlier found to neutralize bioreactive compounds and to be rich in eggs. Accordingly, it is suggested that 28GST plays important roles in phase II defense system and physiological roles in worm fecundity of C. sinensis.
Animals
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Clonorchis sinensis/*enzymology
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Glutathione Transferase/*metabolism/physiology
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Immunohistochemistry
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Microscopy, Immunoelectron
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Molecular Weight
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Support, Non-U.S. Gov't
5.Effects of Excretory/Secretory Products from Clonorchis sinensis and the Carcinogen Dimethylnitrosamine on the Proliferation and Cell Cycle Modulation of Human Epithelial HEK293T Cells.
Eun Min KIM ; June Sung KIM ; Min Ho CHOI ; Sung Tae HONG ; Young Mee BAE
The Korean Journal of Parasitology 2008;46(3):127-132
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.
Animals
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Carcinogens/*metabolism/*toxicity
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Cell Cycle/drug effects
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Cell Line
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Cell Proliferation
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Clonorchis sinensis/*metabolism
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Dimethylnitrosamine/*toxicity
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Epithelial Cells/*drug effects
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Humans
6.Pathological Lesions and Inducible Nitric Oxide Synthase Expressions in the Liver of Mice Experimentally Infected with Clonorchis sinensis.
Qing Li YANG ; Ji Qing SHEN ; Yan XUE ; Xiao Bing CHENG ; Zhi Hua JIANG ; Yi Chao YANG ; Ying Dan CHEN ; Xiao Nong ZHOU
The Korean Journal of Parasitology 2015;53(6):777-783
The nitric oxide (NO) formation and intrinsic nitrosation may be involved in the possible mechanisms of liver fluke-associated carcinogenesis. We still do not know much about the responses of inducible NO synthase (iNOS) induced by Clonorchis sinensis infection. This study was conducted to explore the pathological lesions and iNOS expressions in the liver of mice with different infection intensity levels of C. sinensis. Extensive periductal inflammatory cell infiltration, bile duct hyperplasia, and fibrosis were commonly observed during the infection. The different pathological responses in liver tissues strongly correlated with the infection intensity of C. sinensis. Massive acute spotty necrosis occurred in the liver parenchyma after a severe infection. The iNOS activity in liver tissues increased, and iNOS-expressing cells with morphological differences were observed after a moderate or severe infection. The iNOS-expressing cells in liver tissues had multiple origins.
Animals
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Clonorchiasis/*enzymology/genetics/parasitology/*pathology
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Clonorchis sinensis/*physiology
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Female
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Humans
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Liver/*enzymology/parasitology/pathology
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Mice
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Mice, Inbred BALB C
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Nitric Oxide Synthase Type II/*genetics/metabolism
7.Changing Patterns of Serum and Bile Antibodies in Re-infected Rats with Clonorchis sinensis.
Hongman ZHANG ; Byung Suk CHUNG ; Shunyu LI ; Min Ho CHOI ; Sung Tae HONG
The Korean Journal of Parasitology 2008;46(1):17-22
Rats develop strong resistance to re-infection and super-infection by Clonorchis sinensis. The present study investigated the antibodies present in the sera and bile juice of rats that were primary infected and re-infected with C. sinensis. The serum level of specific IgG antibodies, which were elevated 2 wk of the primary infection, peaked at 4 wk and subsequently remained unchanged even during re-infection. The total IgE level in serum increased slowly from 388 ng / ml to 3,426 ng / ml beginning 2 wk after the primary infection, and remained high up to 8 wk but dropped to a normal level (259 ng / ml) after treatment. In resistant re-infected rats, the serum IgE level increased rapidly and peaked within 1 wk, whereas no increase was observed in immunosuppressed rats. The serum level of specific IgA antibodies was elevated beginning 1 wk after infection, and decreased 4 wk after treatment. The total bile IgA level unchanged during the primary infection but increased in treated and re-infected rats. The elevated levels of serum IgE and bile IgA indicate that these immunoglobulins may be correlated with the development of resistance to re-infection by C. sinensis in rats.
Animals
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Antibodies, Helminth/analysis/*blood/*immunology/metabolism
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Bile/*immunology
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Clonorchiasis/blood/*immunology
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Clonorchis sinensis/*immunology/*physiology
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Immunoglobulin A/analysis/blood
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Male
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Rats
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Rats, Sprague-Dawley
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Time Factors
8.Changing Patterns of Serum and Bile Antibodies in Re-infected Rats with Clonorchis sinensis.
Hongman ZHANG ; Byung Suk CHUNG ; Shunyu LI ; Min Ho CHOI ; Sung Tae HONG
The Korean Journal of Parasitology 2008;46(1):17-22
Rats develop strong resistance to re-infection and super-infection by Clonorchis sinensis. The present study investigated the antibodies present in the sera and bile juice of rats that were primary infected and re-infected with C. sinensis. The serum level of specific IgG antibodies, which were elevated 2 wk of the primary infection, peaked at 4 wk and subsequently remained unchanged even during re-infection. The total IgE level in serum increased slowly from 388 ng / ml to 3,426 ng / ml beginning 2 wk after the primary infection, and remained high up to 8 wk but dropped to a normal level (259 ng / ml) after treatment. In resistant re-infected rats, the serum IgE level increased rapidly and peaked within 1 wk, whereas no increase was observed in immunosuppressed rats. The serum level of specific IgA antibodies was elevated beginning 1 wk after infection, and decreased 4 wk after treatment. The total bile IgA level unchanged during the primary infection but increased in treated and re-infected rats. The elevated levels of serum IgE and bile IgA indicate that these immunoglobulins may be correlated with the development of resistance to re-infection by C. sinensis in rats.
Animals
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Antibodies, Helminth/analysis/*blood/*immunology/metabolism
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Bile/*immunology
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Clonorchiasis/blood/*immunology
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Clonorchis sinensis/*immunology/*physiology
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Immunoglobulin A/analysis/blood
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Male
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Rats
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Rats, Sprague-Dawley
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Time Factors
9.Excretory/secretory antigens from Clonorchis sinensis induces hepatic fibrosis in rats.
Feng-Yu HU ; Xu-Chu HU ; Chang-Ling MA ; Jin XU ; Xin-Bing YU
Journal of Southern Medical University 2009;29(3):393-396
OBJECTIVETo investigate the role of excretory/secretory antigens from Clonorchis sinensis (CsESAs) in hepatic fibrosis induced by C. sinensis infection in rats and explore the possible mechanism.
METHODSCsESAs was collected from adult C. sinensis cultured in sterile condition for 12 h and injected intraperitoneally in Wistar rats. Masson staining was used to observe the changes in the hepatic collagen fiber after the injection. HE staining and immunofluorescence staining were performed to detect the expression of alpha-smooth muscle actin (alpha-SMA) to examine the proliferation and the activity of hepatic stellate cells. The specific antibody titer of CsESAs was determined using enzyme-linked immunosorbent assay to investigate the role of the antigen-antibody complex in the development of hepatic fibrosis.
RESULTSAfter intraperitoneal injection of CsESAs, obvious hepatic fibrosis and hepatic stellate cell proliferation and activation were observed in the rat livers. The severity of the hepatic fibrosis was associated with the dose of CsESAs injected, whereas the titer of the specific antibody against CsESAs showed no direct relation to the hepatic fibrosis.
CONCLUSIONIntraperitoneal injection of CsESAs can cause hepatic stellate cell activation and hepatic fibrosis in rats, but the antigen-antibody complex does not seem to play the key role in the activation of the hepatic stellate cells.
Actins ; metabolism ; Animals ; Antigens, Helminth ; immunology ; Clonorchiasis ; parasitology ; Clonorchis sinensis ; immunology ; pathogenicity ; Hepatic Stellate Cells ; pathology ; Liver Cirrhosis ; immunology ; parasitology ; Male ; Rats ; Rats, Wistar
10.Cloning of a pore-forming subunit of ATP-sensitive potassium channel from Clonorchis sinensis.
Seung Young HWANG ; Hye Jin HAN ; So Hee KIM ; Sae Gwang PARK ; Dae Hyun SEOG ; Na Ri KIM ; Jin HAN ; Joon Yong CHUNG ; Weon Gyu KHO
The Korean Journal of Parasitology 2003;41(2):129-133
A complete cDNA sequence encoding a pore-forming subunit (Kir6.2) of ATP-senstive potassium channel in the adult worm, Clonorchis sinensis, termed CsKir6.2, was isolated from an adult cDNA library. The cDNA contained a single open-reading frame of 333 amino acids, which has a structural motif (a GFG-motif) of the putative pore-forming loop of the Kir6.2. Peculiarly, the CsKir6.2 shows a lack-sequence structure, which deleted 57 amino acids were deleted from its N-terminus. The predicted amino acid sequence revealed a highly conserved sequence as other known other Kir6.2 subunits. The mRNA was weekly expressed in the adult worm.
Adenosine Triphosphate/metabolism
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Amino Acid Sequence
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Animals
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Base Sequence
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Cloning, Molecular
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Clonorchis sinensis/*genetics/metabolism
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Helminth Proteins/*genetics/metabolism
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Human
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Molecular Sequence Data
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Potassium Channels, Inwardly Rectifying/*genetics/metabolism
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RNA, Helminth/chemistry/genetics
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Sequence Alignment
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Support, Non-U.S. Gov't