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Takeshi Agatsuma - One of the best experts on this subject based on the ideXlab platform.
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gene structure of the two domain taurocyamine kinase from Paragonimus Westermani evidence for a distinct lineage of trematode phosphagen kinases
FEBS Letters, 2013Co-Authors: Mitsuru Nagataki, Blanca R Jarilla, Shinji Tokuhiro, Tomohiko Suzuki, Luz P Acosta, Takeshi AgatsumaAbstract:Taurocyamine kinase (TK) is an enzyme that catalyzes the reversible transfer of a phosphate between ATP and taurocyamine. Annelid TKs were suggested to have evolved from a CK ancestor. However, TKs from the lung fluke Paragonimus Westermani comprised another lineage. Construction of phylogenetic tree and comparison of exon/intron organization showed that P. Westermani TK and other trematode TKs evolved from a molluscan arginine kinase (AK) gene. Exon shuffling probably caused the changes in amino acid sequence thereby changing the affinity from AK to TK. The present study provides new insights on the evolution of phosphagen kinases found in trematodes.
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presence of three distinct genotypes within the Paragonimus Westermani complex in northeastern india
Parasitology, 2013Co-Authors: Rekha K Devi, David Blair, Kanwar Narain, Jagadish Mahanta, Tulika Nirmolia, Sidhartha Protim Saikia, Takeshi AgatsumaAbstract:The name Paragonimus Westermani (Kerbert, 1878) is commonly applied to members of a species complex that includes the well-known Asian lung fluke of medical and veterinary importance. Unambiguous molecular and morphological evidence showing the presence of a member of the complex in India has recently been published. In the present study we report the occurrence of 2 more members of the P. Westermani complex in northeastern (NE) India. Surveys of the freshwater crabs Maydelliatelphusa lugubris in NE India revealed 2 morphologically distinct types of lung fluke metacercariae. Phylogenetic analyses, using DNA sequences from ITS2, 28S and cox1 gene regions indicate that these lung metacercariae belong to P. Westermani complex. Type 1 metacercariae have a more basal position within the complex whereas type 2 metacercariae are closely related to the relatively derived forms of P. Westermani from NE Asia (Japan, Korea, China) and Vietnam. A third type of metacercaria (type 3), detected in another crab host, Sartoriana spinigera in Assam, was phylogenetically close to P. siamensis, also a member of the P. Westermani group. Molecular evidence has demonstrated the existence of 3 genotypes of lung flukes within the Paragonimus Westermani complex in NE India. Two of these were previously unknown.
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morphological and molecular characterization of Paragonimus Westermani in northeastern india
Acta Tropica, 2010Co-Authors: Rekha K Devi, David Blair, Lalani Yatawara, Takeshi Agatsuma, Kanwar Narain, Mitsuru Nagataki, Susiji Wickramasinghe, Jagadish MahantaAbstract:Evidence for the presence of lung flukes of the Paragonimus Westermani in India remains scant. In particular, evidence based on morphology of adult worms is lacking. Metacercariae of the genus Paragonimus, recovered from crabs in two regions of northeastern India, were raised to adulthood in laboratory rats. Morphologically, these worms appear to be P. Westermani. DNA sequences from the second internal transcribed spacer (ITS2) and a portion of the ribosomal large subunit gene (28S) of the nuclear ribosomal RNA gene repeat, as well as fragments of the mitochondrial cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase subunit 1 (nad1) genes, all supported this identification. Molecular phylogenetic methods were used for studying the relatedness of these Indian flukes with counterparts from southeast and far-east Asia. Molecular data showed that Indian representatives of the P. Westermani complex represent a distinct lineage. It is unclear whether the Indian form can cause disease in humans as some members of the complex do elsewhere.
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molecular characterization and kinetic properties of a novel two domain taurocyamine kinase from the lung fluke Paragonimus Westermani
FEBS Letters, 2009Co-Authors: Blanca R Jarilla, Sungjong Hong, Mitsuru Nagataki, Shinji Tokuhiro, Tomohiko Suzuki, Takeshi AgatsumaAbstract:Taurocyamine kinase (TK) was previously reported to be restricted to certain marine annelids; however, the present study has proven otherwise. The lung fluke Paragonimus Westermani has a contiguous two-domain TK with a mass of 80 216 Da consisting of 713 amino acid residues sharing higher sequence identity with molluscan arginine kinase (AK). Both domains of P. Westermani TK have significant activity for the substrate taurocyamine and exhibited synergism during substrate binding. Since TK plays a key role in energy metabolism and is not present in mammals, inhibitors against P. Westermani TK could be effective novel chemotherapeutic agents and could be utilized for the development of specific diagnostic tools for the detection of paragonimiasis.
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the first intermediate host of Paragonimus Westermani in sri lanka
Acta Tropica, 2009Co-Authors: Moritoshi Iwagami, R Jayanthe P V Rajapakse, Lalani Yatawara, Shigeyuki Kano, Takeshi AgatsumaAbstract:Abstract Freshwater snails (family Paludomidae, genus Paludomus ) were collected from streams in Hedeniya and Peradeniya (the campus of Peradeniya University), Kandy district, Central Province, Sri Lanka, and found to harbor rediae and cercariae of a Paragonimus sp. These larvae were identified as Paragonimus Westermani by using ITS2 DNA sequences. The infection rates of P. Westermani in Paludomus sp. in Hedeniya and Peradeniya were 0.1% (one of 1014) and 0.2% (two of 1006), respectively. The snail has not been identified to species in the present study. This is the first report of the snail host of Paragonimus in Sri Lanka.
Yukifumi Nawa - One of the best experts on this subject based on the ideXlab platform.
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molecular and morphological variation of Paragonimus Westermani in vietnam with records of new second intermediate crab hosts and a new locality in a northern province
Parasitology, 2016Co-Authors: Pham N Doanh, David Blair, Yoichiro Horii, Luu A Tu, Ho T Loan, Nariaki Nonaka, Yukifumi NawaAbstract:Paragonimus Westermani is one of the most medically important lung flukes and is widely distributed in Asia. It exhibits considerable variation in morphological, genetic and biological features. In central provinces of Vietnam, a high prevalence of metacercariae of this species has been reported from the crab intermediate host, Vietopotamon aluoiense. In this study, we detected P. Westermani metacercariae in two additional crab hosts, Donopotamon haii in Quang Tri Province, central Vietnam and Indochinamon tannanti in Yen Bai Province in the north. The latter is a new locality for P. Westermani in a northern region of Vietnam where P. heterotremus is the only species currently known to cause human paragonimiasis. Paragonimus Westermani metacercariae found in Vietnam showed considerable morphological variation but slight genetic variation based on DNA sequences from the nuclear ribosomal ITS2 region and the mitochondrial 16S gene. Co-infection of the same individual crabs with P. Westermani and P. heterotremus and/or some other Paragonimus species was found frequently, suggesting potential for co-infection in humans. The findings of the present study emphasize the need for highly specific molecular and immunodiagnostic methods to differentially diagnose between P. Westermani and P. heterotremus infections.
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Paragonimus Westermani and Paragonimus species
2014Co-Authors: Yukifumi Nawa, Urusa Thaenkham, Ngoc P Doanh, David BlairAbstract:Paragonimiasis is a zoonotic food-borne lung disease caused by lung flukes of the genus Paragonimus and acquired by consumption of raw/undercooked freshwater crabs/crayfish or wild boar meat. Paragonimus Westermani is the best known species to infect humans in Asia, but several other species of human pathogens are also present. Molecular phylogenetic analyses assign most Asian Paragonimus species into four species complexes with some correlation to human pathogenesis. Paragonimus species exploit a range of mammalian definitive hosts and, as intermediate hosts, freshwater snails and crustaceans, eradication of which is not feasible. Avoidance of consumption of raw/undercooked foods and early diagnosis/treatment are recommended for the control of this disease.
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helminth trematode Paragonimus Westermani and Paragonimus species
Reference Module in Food Science#R##N#Encyclopedia of Food Safety, 2014Co-Authors: Yukifumi Nawa, Urusa Thaenkham, Ngoc P Doanh, David BlairAbstract:Paragonimiasis is a zoonotic food-borne lung disease caused by lung flukes of the genus Paragonimus and acquired by consumption of raw/undercooked freshwater crabs/crayfish or wild boar meat. Paragonimus Westermani is the best known species to infect humans in Asia, but several other species of human pathogens are also present. Molecular phylogenetic analyses assign most Asian Paragonimus species into four species complexes with some correlation to human pathogenesis. Paragonimus species exploit a range of mammalian definitive hosts and, as intermediate hosts, freshwater snails and crustaceans, eradication of which is not feasible. Avoidance of consumption of raw/undercooked foods and early diagnosis/treatment are recommended for the control of this disease.
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infection of a group of boar hunting dogs with Paragonimus Westermani in miyazaki prefecture japan
Veterinary Parasitology, 2008Co-Authors: Yumi Kirino, Yukifumi Nawa, Fukumi Nakamurauchiyama, Natsuko Nakano, Mitsuyoshi Hagio, Yuichi Hidaka, Yoichiro HoriiAbstract:A 5-year-old male mixed-bred boar-hunting dog with a Plott hound background weighing 23 kg was brought to the Veterinary Teaching Hospital, University of Miyazaki, in October 2002. The dog was diagnosed with active infection with the lung fluke Paragonimus Westermani by serological testing and also by detection of parasite eggs in his feces. Subsequent examination of four other dogs working with this dog as a boar-hunting team revealed that all five dogs were infected with P. Westermani.
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active hepatic capsulitis caused by Paragonimus Westermani infection
Internal Medicine, 2002Co-Authors: Michiro Sasaki, Takuro Kamiyama, Takeshi Yano, Fukumi Nakamurauchiyama, Yukifumi NawaAbstract:Paragonimiasis is an important re-emerging parasitosis in Japan. Although the lungs and pleural cavity are the principal sites affected with the parasite, ectopic infection can occur in unexpected sites such as skin and brain. This case report describes a patient with active hepatic capsulitis due to Paragonimus Westermani infection. The patient was successfully treated with praziquantel at the dose of 75 mg/kg/day for 3 days.(Internal Medicine 41: 661-663, 2002)
David Blair - One of the best experts on this subject based on the ideXlab platform.
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molecular and morphological variation of Paragonimus Westermani in vietnam with records of new second intermediate crab hosts and a new locality in a northern province
Parasitology, 2016Co-Authors: Pham N Doanh, David Blair, Yoichiro Horii, Luu A Tu, Ho T Loan, Nariaki Nonaka, Yukifumi NawaAbstract:Paragonimus Westermani is one of the most medically important lung flukes and is widely distributed in Asia. It exhibits considerable variation in morphological, genetic and biological features. In central provinces of Vietnam, a high prevalence of metacercariae of this species has been reported from the crab intermediate host, Vietopotamon aluoiense. In this study, we detected P. Westermani metacercariae in two additional crab hosts, Donopotamon haii in Quang Tri Province, central Vietnam and Indochinamon tannanti in Yen Bai Province in the north. The latter is a new locality for P. Westermani in a northern region of Vietnam where P. heterotremus is the only species currently known to cause human paragonimiasis. Paragonimus Westermani metacercariae found in Vietnam showed considerable morphological variation but slight genetic variation based on DNA sequences from the nuclear ribosomal ITS2 region and the mitochondrial 16S gene. Co-infection of the same individual crabs with P. Westermani and P. heterotremus and/or some other Paragonimus species was found frequently, suggesting potential for co-infection in humans. The findings of the present study emphasize the need for highly specific molecular and immunodiagnostic methods to differentially diagnose between P. Westermani and P. heterotremus infections.
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Paragonimus Westermani and Paragonimus species
2014Co-Authors: Yukifumi Nawa, Urusa Thaenkham, Ngoc P Doanh, David BlairAbstract:Paragonimiasis is a zoonotic food-borne lung disease caused by lung flukes of the genus Paragonimus and acquired by consumption of raw/undercooked freshwater crabs/crayfish or wild boar meat. Paragonimus Westermani is the best known species to infect humans in Asia, but several other species of human pathogens are also present. Molecular phylogenetic analyses assign most Asian Paragonimus species into four species complexes with some correlation to human pathogenesis. Paragonimus species exploit a range of mammalian definitive hosts and, as intermediate hosts, freshwater snails and crustaceans, eradication of which is not feasible. Avoidance of consumption of raw/undercooked foods and early diagnosis/treatment are recommended for the control of this disease.
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helminth trematode Paragonimus Westermani and Paragonimus species
Reference Module in Food Science#R##N#Encyclopedia of Food Safety, 2014Co-Authors: Yukifumi Nawa, Urusa Thaenkham, Ngoc P Doanh, David BlairAbstract:Paragonimiasis is a zoonotic food-borne lung disease caused by lung flukes of the genus Paragonimus and acquired by consumption of raw/undercooked freshwater crabs/crayfish or wild boar meat. Paragonimus Westermani is the best known species to infect humans in Asia, but several other species of human pathogens are also present. Molecular phylogenetic analyses assign most Asian Paragonimus species into four species complexes with some correlation to human pathogenesis. Paragonimus species exploit a range of mammalian definitive hosts and, as intermediate hosts, freshwater snails and crustaceans, eradication of which is not feasible. Avoidance of consumption of raw/undercooked foods and early diagnosis/treatment are recommended for the control of this disease.
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presence of three distinct genotypes within the Paragonimus Westermani complex in northeastern india
Parasitology, 2013Co-Authors: Rekha K Devi, David Blair, Kanwar Narain, Jagadish Mahanta, Tulika Nirmolia, Sidhartha Protim Saikia, Takeshi AgatsumaAbstract:The name Paragonimus Westermani (Kerbert, 1878) is commonly applied to members of a species complex that includes the well-known Asian lung fluke of medical and veterinary importance. Unambiguous molecular and morphological evidence showing the presence of a member of the complex in India has recently been published. In the present study we report the occurrence of 2 more members of the P. Westermani complex in northeastern (NE) India. Surveys of the freshwater crabs Maydelliatelphusa lugubris in NE India revealed 2 morphologically distinct types of lung fluke metacercariae. Phylogenetic analyses, using DNA sequences from ITS2, 28S and cox1 gene regions indicate that these lung metacercariae belong to P. Westermani complex. Type 1 metacercariae have a more basal position within the complex whereas type 2 metacercariae are closely related to the relatively derived forms of P. Westermani from NE Asia (Japan, Korea, China) and Vietnam. A third type of metacercaria (type 3), detected in another crab host, Sartoriana spinigera in Assam, was phylogenetically close to P. siamensis, also a member of the P. Westermani group. Molecular evidence has demonstrated the existence of 3 genotypes of lung flukes within the Paragonimus Westermani complex in NE India. Two of these were previously unknown.
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morphological and molecular characterization of Paragonimus Westermani in northeastern india
Acta Tropica, 2010Co-Authors: Rekha K Devi, David Blair, Lalani Yatawara, Takeshi Agatsuma, Kanwar Narain, Mitsuru Nagataki, Susiji Wickramasinghe, Jagadish MahantaAbstract:Evidence for the presence of lung flukes of the Paragonimus Westermani in India remains scant. In particular, evidence based on morphology of adult worms is lacking. Metacercariae of the genus Paragonimus, recovered from crabs in two regions of northeastern India, were raised to adulthood in laboratory rats. Morphologically, these worms appear to be P. Westermani. DNA sequences from the second internal transcribed spacer (ITS2) and a portion of the ribosomal large subunit gene (28S) of the nuclear ribosomal RNA gene repeat, as well as fragments of the mitochondrial cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase subunit 1 (nad1) genes, all supported this identification. Molecular phylogenetic methods were used for studying the relatedness of these Indian flukes with counterparts from southeast and far-east Asia. Molecular data showed that Indian representatives of the P. Westermani complex represent a distinct lineage. It is unclear whether the Indian form can cause disease in humans as some members of the complex do elsewhere.
Yoon Kong - One of the best experts on this subject based on the ideXlab platform.
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Paragonimus Westermani biochemical and immunological characterizations of paramyosin
Experimental Parasitology, 2007Co-Authors: Yoon Kong, Qin Ping Zhao, Sungung Moon, Byoungkuk Na, Woonmok Sohn, Mingsen JiangAbstract:Paramyosin of the helminth parasite is a muscle protein that plays multifunctional roles in host-parasite relationships. In this study, we have cloned a gene encoding Paragonimus Westermani paramyosin (PwPmy) and characterized biochemical and immunological properties of the recombinant protein. The recombinant PwPmy (rPwPmy) was shown to bind both human immunoglobulin G (IgG) and collagen. The protein was constitutively expressed in various developmental stages of the parasite and its expression level increased progressively as the parasite matured. Immunohistological analysis revealed that PwPmy was mainly localized in subtegumental muscle, tegument and cells surrounding the oral sucker, intestine, and ovary of the parasite. Sera from patients with paragonimiasis showed antibody reactivity against rPwPmy, and IgG1 and IgG4 were predominant. Immunization of mice with rPwPmy also induced high IgG responses. Biochemical and immunological characterization of PwPmy may provide valuable information for the further study to develop a vaccine or a chemotherapeutic agent for paragonimiasis.
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molecular cloning and characterization of copper zinc superoxide dismutase of Paragonimus Westermani
Journal of Parasitology, 2005Co-Authors: Aihua Li, Yoon Kong, Byoungkuk Na, Qin Ping ZhaoAbstract:Superoxide dismutases (SODs; EC 1.15.1.1) play important roles in the protection of the parasites against cellular oxygen–mediated killing of the hosts. A copper/zinc-containing SOD (Cu/Zn-SOD) was identified previously from lung fluke, Paragonimus Westermani. To expand our understanding of P. Westermani SOD, we isolated a complementary DNA encoding a Cu/Zn-SOD, expressed the active enzyme in Escherichia coli, and characterized its biochemical properties. The deduced amino acid (aa) sequence of the gene shared up to 73.7% identities with Cu/Zn-SODs of other helminths and shared well-conserved characteristic motifs and essential aa residues involved in coordinating copper and zinc enzymatic functions. Recombinant Cu/ Zn-SOD exhibited comparable biochemical properties with that of the native enzyme, including pH optima and potassium cyanide–and hydrogen peroxide–sensitive inhibition profiles. The active enzyme consisted of 2 identical subunits covalently linked by disulfide bonds. The enzyme was constitutiv...
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divergent long terminal repeat retrotransposon families in the genome of Paragonimus Westermani
Korean Journal of Parasitology, 2003Co-Authors: Yoon KongAbstract:To gain information on retrotransposons in the genome of Paragonimus Westermani, PCR was carried out with degenerate primers, specific to protease and reverse transcriptase (rt) genes of long-terminal-repeat (LTR) retrotransposons. The PCR products were cloned and sequenced, after which 12 different retrotransposon-related sequences were isolated from the trematode genome. These showed various degrees of identity to the polyprotein of divergent retrotransposon families. A phylogenetic analysis demonstrated that these sequences could be classified into three different families of LTR retrotransposons, namely, Xena, Bel, and Gypsy families. Of these, two mRNA transcripts were detected by reverse transcriptase-PCR, showing that these two elements preserved their mobile activities. The genomic distributions of these two sequences were found to be highly repetitive. These results suggest that there are diverse retrotransposons including the ancient Xena family in the genome of P. Westermani, which may have been involved in the evolution of the host genome.
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excretory bladder the source of cysteine proteases in Paragonimus Westermani metacercariae
Korean Journal of Parasitology, 2002Co-Authors: Hyunjong Yang, Shinyong Kang, Y B Chung, Yoon KongAbstract:The cysteine proteases of Paragonimus Westermani metacercariae are involved in metacercarial excystment, host immune modulation, and possibly in tissue penetration. In order to clarify the origin of the enzymes, 28 and 27 kDa cysteine proteases in metacercarial excretory-secretory products were purified through the FPLC system using Mono Q column chromatography. The polyclonal antibodies to the enzymes were produced in BALB/c mice. Immunolocalization studies revealed that both cysteine proteases were distributed at the linings of excretory bladder and excretory concretions of the metacercariae. It was suggested that the excretory epithelium of P. Westermani undertake the secretory function of metacercarial cysteine proteases, in addition to its role as a route for eliminating waste products.
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recombinant Paragonimus Westermani yolk ferritin is a useful serodiagnostic antigen
The Journal of Infectious Diseases, 2002Co-Authors: Shinyong Kang, Yoon Kong, Kom Sukomtason, Sungjong HongAbstract:A recombinant protein of Paragonimus Westermani yolk ferritin was bacterially produced from a previously cloned complementary DNA and was used as an antigen for an enzyme-linked immunosorbent assay (ELISA) against paragonimiasis- and other helminth-infected sera to evaluate its serodiagnostic potential. The ELISA revealed that paragonimiasis Westermani had 88.2% sensitivity and 100% specificity. The positive and negative predictive values of the ELISA were calculated to be 100% and 97.1%, respectively. Sera from cats experimentally infected with P. Westermani began to produce immunoglobulin G antibodies against the yolk ferritin at 13 weeks after infection, which suggests that the corresponding antigen was derived from the vitellaria in accordance with maturation of P. Westermani. These results indicate that the recombinant P. Westermani yolk ferritin is a potent serodiagnostic reagent for paragonimiasis Westermani from an early stage of the infection.
Sungjong Hong - One of the best experts on this subject based on the ideXlab platform.
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inactivation of Paragonimus Westermani metacercariae in soy sauce marinated and frozen freshwater crabs
Parasitology Research, 2017Co-Authors: Sera Oh, Hyunjong Yang, Sangdo Ha, Sungjong HongAbstract:Soy sauce-marinated freshwater crabs (Eriocheir japonicus) are a source of human paragonimiasis. The viability of Paragonimus Westermani metacercariae (PwMc) in marinated crabs was investigated in an experimental setting. The PwMc collected from freshwater crayfish were inoculated into freshwater crabs, which were then frozen or marinated in soy sauce. All PwMc in the freshwater crabs were inactivated after freezing for 48 h at −20 °C and after freezing for 12 h at −40 °C. After marinating for 32 days, the survival rate of PwMc in 5% NaCl soy sauce was 50%, in 7.5% NaCl soy sauce it was 33.3%, and in 10.0% NaCl soy sauce it was 31.3%. When marinated for 64 days, all PwMc were inactivated in all experimental groups. These results revealed that freezing and soy sauce marination were detrimental to the survival of PwMc in freshwater crabs. Specifically, freezing crabs for more than 48 h or soaking them in soy sauce containing at least 5.0% NaCl for 64 days can inactivate PwMc. These results can inform the production of the traditional Korean soy sauce-marinated freshwater crabs known as gejang.
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molecular characterization and kinetic properties of a novel two domain taurocyamine kinase from the lung fluke Paragonimus Westermani
FEBS Letters, 2009Co-Authors: Blanca R Jarilla, Sungjong Hong, Mitsuru Nagataki, Shinji Tokuhiro, Tomohiko Suzuki, Takeshi AgatsumaAbstract:Taurocyamine kinase (TK) was previously reported to be restricted to certain marine annelids; however, the present study has proven otherwise. The lung fluke Paragonimus Westermani has a contiguous two-domain TK with a mass of 80 216 Da consisting of 713 amino acid residues sharing higher sequence identity with molluscan arginine kinase (AK). Both domains of P. Westermani TK have significant activity for the substrate taurocyamine and exhibited synergism during substrate binding. Since TK plays a key role in energy metabolism and is not present in mammals, inhibitors against P. Westermani TK could be effective novel chemotherapeutic agents and could be utilized for the development of specific diagnostic tools for the detection of paragonimiasis.
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Paragonimus Westermani molecular cloning expression and characterization of a recombinant yolk ferritin
Experimental Parasitology, 2002Co-Authors: Shinyong Kang, Sungjong HongAbstract:Abstract Ferritin is an intracellular protein involved in iron metabolism. A cDNA PwYF-1 cloned from the adult Paragonimus Westermani cDNA library encoded a putative polypeptide of 216 amino acids homologous with ferritins of vertebrates and invertebrates. Fe-binding motifs identified in PwYF-1 polypeptide were conserved and predicted to form a ferroxidase center. PwYF-1 polypeptide contained an extended peptide of 45 amino acids at its C-terminus. Recombinant PwYF-1 protein, expressed and purified from Escherichia coli, showed iron-uptake ability and ferroxidase activity. Ferroxidase activity of recombinant PwYF-1 protein was reactivated by secondary addition of apotransferrin to assay mixture. Mouse immune serum raised against the recombinant PwYF-1 protein recognized specifically 24 kDa protein from adult P. Westermani lysate. PwYF-1 protein was localized to vitelline follicles and the eggs of P. Westermani. Collectively, PwYF-1 protein was identified as a P. Westermani yolk ferritin. Abbreviations: PwYF-1, Paragonimus Westermani yolk ferritin-1; PCR, polymerase chain reaction; MTPBS, mouse tonicity phosphate-buffered saline; Ni–NTA, nickel–nitrilotriacetic acid
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recombinant Paragonimus Westermani yolk ferritin is a useful serodiagnostic antigen
The Journal of Infectious Diseases, 2002Co-Authors: Shinyong Kang, Yoon Kong, Kom Sukomtason, Sungjong HongAbstract:A recombinant protein of Paragonimus Westermani yolk ferritin was bacterially produced from a previously cloned complementary DNA and was used as an antigen for an enzyme-linked immunosorbent assay (ELISA) against paragonimiasis- and other helminth-infected sera to evaluate its serodiagnostic potential. The ELISA revealed that paragonimiasis Westermani had 88.2% sensitivity and 100% specificity. The positive and negative predictive values of the ELISA were calculated to be 100% and 97.1%, respectively. Sera from cats experimentally infected with P. Westermani began to produce immunoglobulin G antibodies against the yolk ferritin at 13 weeks after infection, which suggests that the corresponding antigen was derived from the vitellaria in accordance with maturation of P. Westermani. These results indicate that the recombinant P. Westermani yolk ferritin is a potent serodiagnostic reagent for paragonimiasis Westermani from an early stage of the infection.
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Paragonimus Westermani a cytosolic glutathione s transferase of a σ class in adult stage
Experimental Parasitology, 2000Co-Authors: Sungjong Hong, Shinyong Kang, Y B Chung, Minhyun Chung, Younjin Oh, Insug Kang, Young Yil Bahk, Yoon KongAbstract:Abstract Hong, S.-J., Kang, S.-Y., Chung, Y.-B., Chung, M.-H., Oh, Y.-J., Kang, I., Bahk, Y.Y., Kong, Y., and Cho, S.-Y. 2000. Paragonimus Westermani: A cytosolic glutathione S -transferase of a σ-class in adult stage. Experimental Parasitology 94, 180–189. We purified cytosolic glutathione S -transferase (GST) of adult Paragonimus Westermani monitoring its activity with 1-chloro-2,4-dinitrobenzene (CDNB). The enzyme was purified 18.4-fold to electrophoretic homogeneity with 21% recovery rate through a three-step procedure. The purified enzyme (Pw28GST) has a subunit molecular weight of 28 kDa with an isoelectric point at 4.6. Monoclonal antibody (anti-Pw28GST) against Pw28GST did not cross-react with GSTs from other helminths. cDNA library was constructed in λZAP II bacteriophage and screened with anti-Pw28GST. The corresponding gene containing a single open reading frame of 804 bp encoded 211 amino acids. The predicted amino acid sequence exhibited a higher homology with catalytic domain near N-terminus of class σ GSTs (58%) than with schistosome 28-kDa GSTs (45–41%) or with class σ GSTs themselves (33–31%). The sequence contained both Tyr-6 and Tyr-10 that are highly conserved in mammalian and helminth GSTs. The apparent K m value of a recombinant enzyme was 0.78 mM. Both native and recombinant enzymes showed the highest activity against CDNB, relatively weak activity against ethacrynic acid and reactive carbonyls, and no activity against epoxy-3-( p -nitrophenoxy)-propane. The activities were inhibited by bromosulfophthalein, cibacron blue, and albendazole, but not by praziquantel. These findings indicate that adult P. Westermani has a class σ GST.
