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Internal Medicine (Tokyo, Japan) Aug 2020An asymptomatic 47-year-old woman was admitted with pleural effusion and pulmonary infiltrates 1 month after ingesting raw wild boar and deer meat. Both her blood and...
An asymptomatic 47-year-old woman was admitted with pleural effusion and pulmonary infiltrates 1 month after ingesting raw wild boar and deer meat. Both her blood and pleural fluid were eosinophilic. Thoracoscopy revealed multiple nodules of the pleura, and biopsy samples of the nodules showed necrosis with epithelioid cell granulomas. An enzyme-linked immunosorbent assay was positive for antibodies against Paragonimus westermani, and the patient was successfully treated with praziquantel. This is the first reported case of pulmonary or pleuropulmonary paragonimiasis where several pleural nodules were observed. The detection of pleural nodules on thoracoscopy can contribute to the prompt and accurate diagnosis of paragonimiasis.
Topics: Animals; Deer; Enzyme-Linked Immunosorbent Assay; Female; Humans; Meat; Middle Aged; Paragonimiasis; Paragonimus westermani; Pleura; Pleural Effusion; Praziquantel; Respiratory Tract Infections; Sus scrofa; Thoracoscopy
PubMed: 32350198
DOI: 10.2169/internalmedicine.4457-20 -
Clinical and Diagnostic Laboratory... Jul 1998In 40 cases of human paragonimiases caused by Paragonimus westermani (20 cases), P. miyazakii (10 cases), and P. skrjabini (10 cases), responses of serum immunoglobulin...
In 40 cases of human paragonimiases caused by Paragonimus westermani (20 cases), P. miyazakii (10 cases), and P. skrjabini (10 cases), responses of serum immunoglobulin G (IgG), IgG subclasses, and IgE were analyzed by immunoblotting with crude antigens prepared from egg, 4-week-old juvenile, and adult forms of P. westermani. The 32- and 35-kDa proteins in the adult extracts showed specific reactions regardless of the causative species (39 of 40 cases; 98%). Sera of patients infected with P. westermani and P. miyazakii reacted strongly with the 28-, 46-, and 94-kDa proteins of egg extracts, while those from patients infected with P. skrjabini reacted faintly. No sera from patients with other trematodiases (0 of 15 cases), cestodiases (0 of 20 cases), or lung cancer (0 of 5 cases) or from healthy controls (0 of 10 individuals) showed positive reactions. Analysis by IgG subclass revealed that IgG4 (33 of 40 cases; 83%) and IgG1 (29 of 40 cases; 73%) antibodies in the patient sera recognized the 32- and 35-kDa proteins predominantly. IgG3 reaction was found in 50% (10 of 20 cases) and 30% (3 of 10 cases) of the sera of patients infected with P. westermani and P. miyazakii, respectively. In an IgE immunoblot, 83% (33 of 40 cases) of the sera from paragonimiasis patients reacted with the 32- and 35-kDa proteins while no sera from patients with heterologous diseases and healthy controls showed a positive reaction. Both 32- and 35-kDa proteins in adult extracts of P. westermani were highly reliable for serodiagnosis of human paragonimiases.
Topics: Animals; Antibodies, Helminth; Antibody Specificity; Antigens, Helminth; Case-Control Studies; Helminth Proteins; Humans; Immunoblotting; Immunoglobulin E; Immunoglobulin G; Lung Diseases, Parasitic; Molecular Weight; Paragonimiasis; Paragonimus; Sensitivity and Specificity; Serologic Tests; Species Specificity
PubMed: 9665951
DOI: 10.1128/CDLI.5.4.474-478.1998 -
Tropical Medicine and Infectious Disease Dec 2023Human pulmonary paragonimiasis, an emerging concern in North East India, frequently masquerades as pulmonary tuberculosis due to clinical and radiological similarities,...
Human pulmonary paragonimiasis, an emerging concern in North East India, frequently masquerades as pulmonary tuberculosis due to clinical and radiological similarities, leading to diagnostic challenges. This research aimed to harness the immunoblotting technique to discern immunodiagnostic protein antigens from both adult worm and excretory-secretory (ES) extracts of the prevalent type 1 in Arunachal Pradesh, North East India. We studied the time kinetics of immunoreactive patterns in relation to the duration of infection in rodent models. Immunoblot analyses were also conducted using sera from ELISA-positive patients confirmed with paragonimiasis, facilitating the selection of antigenic extracts with diagnostic potential. Further, ES protein antigens were subjected to 2D immunoblot analysis and immunoreactive protein spots identified using MALDI-TOF MS. The immunoreactivity patterns of ES antigens with sera of paragonimiasis-positive patients were detailed, and specific immunoreactive protein antigens were pinpointed using peptide mass fingerprinting (MALDI-TOF). This work underscores the enhanced diagnostic accuracy when combining ELISA with immunoblotting for pulmonary paragonimiasis in regions like North East India, marked by co-existing helminth infections.
PubMed: 38251203
DOI: 10.3390/tropicalmed9010006 -
PeerJ 2014Among helminth parasites, Paragonimus (zoonotic lung fluke) gains considerable importance from veterinary and medical points of view because of its diversified effect on...
Among helminth parasites, Paragonimus (zoonotic lung fluke) gains considerable importance from veterinary and medical points of view because of its diversified effect on its host. Nearly fifty species of Paragonimus have been described across the globe. It is estimated that more than 20 million people are infected worldwide and the best known species is Paragonimus westermani, whose type locality is probably India and which infects millions of people in Asia causing disease symptoms that mimic tuberculosis. Human infections occur through eating raw crustaceans containing metacercarie or ingestion of uncooked meat of paratenic hosts such as pigs. Though the fluke is known to parasitize a wide range of mammalian hosts representing as many as eleven families, the status of its prevalence, host range, pathogenic manifestations and its possible survivors in nature from where the human beings contract the infection is not well documented in India. We took advantage of the whole genome sequence data for P. westermani, generated by Next Generation Sequencing, and its comparison with the existing data for the P. westermani for comparative mt DNA phylogenomic analyses. Specific primers were designed for the 12 protein coding genes with the aid of existing P. westermani mtDNA as the reference. The Ion torrent next generation sequencing platform was harnessed to completely sequence the mitochondrial genome, and applied innovative approaches to bioinformatically assemble and annotate it. A strategic PCR primer design utilizing the whole genome sequence data from P. westermani enabled us to design specific primers capable of amplifying all regions of the mitochondrial genome from P. westermani. Assembly of NGS data from libraries enriched in mtDNA sequence by PCR gave rise to a total of 11 contigs spanning the entire 14.7 kb mt DNA sequence of P. westermani available at NCBI. We conducted gap-filling by traditional Sanger sequencing to fill in the gaps. Annotation of non-protein coding genes successfully identified tRNA regions for the 24 tRNAs coded in mtDNA and 12 protein coding genes. Bayesian phylogenetic analyses of the concatenated protein coding genes placed P. westermani within the family Opisthorchida. The complete mtDNA sequence of P. westermani is 15,004 base pairs long; the lung fluke is the major etiological agent of paragonimiasis and the first Indian representative for the family Paragonimidae to be fully sequenced that provides important genetic markers for ecological, population and biogeographical studies and molecular diagnostic of digeneans that cause trematodiases.
PubMed: 25165620
DOI: 10.7717/peerj.484 -
BMC Genomics Oct 2008Retrotransposons have been known to involve in the remodeling and evolution of host genome. These reverse transcribing elements, which show a complex evolutionary... (Comparative Study)
Comparative Study
PwRn1, a novel Ty3/gypsy-like retrotransposon of Paragonimus westermani: molecular characters and its differentially preserved mobile potential according to host chromosomal polyploidy.
BACKGROUND
Retrotransposons have been known to involve in the remodeling and evolution of host genome. These reverse transcribing elements, which show a complex evolutionary pathway with diverse intermediate forms, have been comprehensively analyzed from a wide range of host genomes, while the information remains limited to only a few species in the phylum Platyhelminthes.
RESULTS
A LTR retrotransposon and its homologs with a strong phylogenetic affinity toward CsRn1 of Clonorchis sinensis were isolated from a trematode parasite Paragonimus westermani via a degenerate PCR method and from an insect species Anopheles gambiae by in silico analysis of the whole mosquito genome, respectively. These elements, designated PwRn1 and AgCR-1 - AgCR-14 conserved unique features including a t-RNATrp primer binding site and the unusual CHCC signature of Gag proteins. Their flanking LTRs displayed >97% nucleotide identities and thus, these elements were likely to have expanded recently in the trematode and insect genomes. They evolved heterogeneous expression strategies: a single fused ORF, two separate ORFs with an identical reading frame and two ORFs overlapped by -1 frameshifting. Phylogenetic analyses suggested that the elements with the separate ORFs had evolved from an ancestral form(s) with the overlapped ORFs. The mobile potential of PwRn1 was likely to be maintained differentially in association with the karyotype of host genomes, as was examined by the presence/absence of intergenomic polymorphism and mRNA transcripts.
CONCLUSION
Our results on the structural diversity of CsRn1-like elements can provide a molecular tool to dissect a more detailed evolutionary episode of LTR retrotransposons. The PwRn1-associated genomic polymorphism, which is substantial in diploids, will also be informative in addressing genomic diversification following inter-/intra-specific hybridization in P. westermani populations.
Topics: Amino Acid Sequence; Animals; Anopheles; DNA, Helminth; Dogs; Drosophila melanogaster; Evolution, Molecular; Genome, Helminth; Genomic Library; Molecular Sequence Data; Open Reading Frames; Paragonimus westermani; Phylogeny; Polymorphism, Genetic; Polyploidy; Retroelements; Reverse Transcriptase Polymerase Chain Reaction; Sequence Analysis, DNA
PubMed: 18851759
DOI: 10.1186/1471-2164-9-482 -
The Journal of Veterinary Medical... Aug 2017Infection of boar-hunting dogs with Paragonimus westermani was investigated in Western Japan. Blood and rectal feces were collected from 441 dogs in the three districts...
Infection of boar-hunting dogs with Paragonimus westermani was investigated in Western Japan. Blood and rectal feces were collected from 441 dogs in the three districts (205 in Kinki, 131 in Chugoku and 105 in Shikoku District). In a screening ELISA for serum antibody against P. westermani antigen, 195 dogs (44.2%) showed positive reaction. In the 195 dogs, 8 dogs were found excreting P. westermani eggs after molecular analysis of fecal eggs, and additional 7 were identified serologically for the parasite infection because of their stronger reactivity against P. westermani antigen than against antigens of other species of Paragonimus. A spatial analysis showed that all of the P. westermani infections were found in Kinki and Chugoku Districts. In this area, dogs' experience of being fed with raw boar meat showed high odds ratio (3.35) to the sero-positivity in the screening ELISA, and the frequency of such experiences was significantly higher in sero-positive dogs. While clear relationship was not obtained between predation of boars by dogs during hunting and their sero-positivity. Therefore, it is suggested that human activity of feeding with wild boar meat is the risk factor for P. westermani infection in boar-hunting dogs. Considering that hunting dogs could play as a major definitive host and maintain the present distribution of P. westermani in Western Japan, control measures for the infection in hunting dogs, such as prohibition of raw meat feeding and regular deworming, should be undertaken.
Topics: Animal Feed; Animals; Dog Diseases; Dogs; Enzyme-Linked Immunosorbent Assay; Japan; Meat; Paragonimiasis; Paragonimus; Sus scrofa; Swine; Swine Diseases
PubMed: 28717056
DOI: 10.1292/jvms.17-0149 -
European Journal of Biochemistry Oct 1995Acid cysteine protease was purified from metacercariae of the mammalian trematode parasite Paragonimus westermani. The purified enzyme had a molecular mass of 27 kDa and...
Acid cysteine protease was purified from metacercariae of the mammalian trematode parasite Paragonimus westermani. The purified enzyme had a molecular mass of 27 kDa and was a monomeric polypeptide. The protease had an absolute requirement for a reducing agent for full activity towards fluorescein-isothiocyanate-labeled hemoglobin, and it was active in the acidic pH range, with an optimum pH of 4.0. While acidic proteolysis was insensitive to the aspartic protease inhibitor pepstatin A, activity was significantly inhibited by the cysteine protease inhibitors, leupeptin, chymostatin and L-trans-epoxy-succinyl-L-leucylamido(4-guanidino)-butane. The sensitivity of the enzyme to the inhibitors was similar to that of cathepsins B and L, but the specificity of the protease towards chromogenic substrates was slightly different from that of the cathepsins. The purified enzyme was highly specific for N-substituted peptidyl substrates containing arginine in the P1 position and phenylalanine in the P2 position, and the protease extensively degraded human native proteins, such as human serum albumin, immunoglobulins, complement components and also endogenous protease inhibitors. Since the protease hydrolyzes both soluble proteins and components of human defense systems, it may facilitate parasite nutrition and evasion of host defense mechanisms.
Topics: Amino Acid Sequence; Animals; Cysteine Endopeptidases; Cysteine Proteinase Inhibitors; Humans; Hydrogen-Ion Concentration; Molecular Sequence Data; Paragonimus
PubMed: 7588793
DOI: 10.1111/j.1432-1033.1995.490_2.x -
The Korean Journal of Parasitology Jun 2017An epidemiological study was performed to know the recent infection status of metacercariae (PwMc) in freshwater crayfish, , from 2 streams in Jeollanam-do, Republic of...
An epidemiological study was performed to know the recent infection status of metacercariae (PwMc) in freshwater crayfish, , from 2 streams in Jeollanam-do, Republic of Korea. Crayfish were collected from creeks in Bogil-do (Island), Wando-gun, and in a creek near Daeheung Temple in Haenam-gun. The infection rate of crayfish with PwMc in Bogil-do was 89.8%, and the metacercarial burden was 37 PwMc per the infected crayfish. Crayfish in a creek near Daeheung Temple were larger and twice heavier than those in Bogil-do. Of them, 96.5% were infected with PwMc. An average of 140 metacercariae was found in the infected crayfish, almost quadruple to those of Bogil-do. There was a strong correlation between the number of PwMc and body weight of the crayfish. These results suggest that metacercariae are still prevalent in crayfish of the 2 regions in Jeollanam-do, Korea.
Topics: Animals; Astacoidea; Body Weight; Fresh Water; Incidence; Metacercariae; Paragonimiasis; Paragonimus westermani; Republic of Korea
PubMed: 28719962
DOI: 10.3347/kjp.2017.55.3.347 -
Journal of Infection and Chemotherapy :... Dec 2016Herein, we report a case of Paragonimus westermani infection, which required differentiation from recurrent lung cancer. A 66-year old Japanese man with a history of...
Herein, we report a case of Paragonimus westermani infection, which required differentiation from recurrent lung cancer. A 66-year old Japanese man with a history of lung cancer who had undergone a lobectomy was referred to our clinic for treatment of cough, sputum, dyspnea, and a right pulmonary nodule. He had previously eaten seafood he visited China. P. westermani infection was confirmed by the presence of antibody against P. westermani antigen in the patient's serum and eggs in his sputum. Eventually, molecular identification by PCR-restriction fragment length polymorphism analysis and sequencing confirmed that the patient was infected with triploid forms of P. westermani.
Topics: Aged; Animals; Humans; Lung Neoplasms; Male; Neoplasm Recurrence, Local; Paragonimiasis; Paragonimus westermani; Sputum
PubMed: 27498617
DOI: 10.1016/j.jiac.2016.07.002 -
Clinical and Diagnostic Laboratory... Nov 2000A complete cDNA sequence encoding a 28-kDa cruzipain-like cysteine protease of adult Paragonimus westermani, termed Pw28CCP, was isolated from an adult cDNA library. The...
A complete cDNA sequence encoding a 28-kDa cruzipain-like cysteine protease of adult Paragonimus westermani, termed Pw28CCP, was isolated from an adult cDNA library. The cDNA contained a single open reading frame of 975 bp encoding 325 amino acids, which exhibited the structural motif and domain organization characteristic of cysteine proteases of non-cathepsin Bs including a hydrophobic signal sequence, an ERFNIN motif, and essential cysteine residues as well as active sites in the mature catalytic region. Analysis of its phylogenetic position revealed that this novel enzyme belonged to the cruzipain-like cysteine proteases. The sequence of the first 13 amino acids predicted from the mature domain of Pw28CCP was in accord with that determined from the native 28-kDa enzyme purified from the adult worm. Expression of Pw28CCP was observed specifically in juvenile and adult worms, with a location in the intestinal epithelium, suggesting that this enzyme could be secreted and involved in nutrient uptake and immune modulation. The recombinant protein expressed in Escherichia coli was used to assess antigenicity by immunoblotting with sera from patients with active paragonimiasis and from those with other parasitic infections. The resulting sensitivity of 86.2% (56 of 65 samples) and specificity of 98% (147 of 150 samples) suggest its potential as an antigen for use in immunodiagnosis.
Topics: Amino Acid Sequence; Animals; Base Sequence; Cats; Cysteine Endopeptidases; DNA Primers; Dogs; Gene Expression Regulation, Developmental; Gene Expression Regulation, Enzymologic; Genes, Helminth; Immunologic Tests; In Situ Hybridization; Molecular Sequence Data; Molecular Weight; Paragonimiasis; Paragonimus; Phylogeny; Protozoan Proteins; Sequence Homology, Amino Acid
PubMed: 11063501
DOI: 10.1128/CDLI.7.6.932-939.2000