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1.
Parasitol Res ; 120(2): 535-545, 2021 Feb.
Article in English | MEDLINE | ID: mdl-33415393

ABSTRACT

Paragonimiasis is a foodborne trematode infection that affects 23 million people, mainly in Asia. Lung fluke infections lead frequently to chronic cough with fever and hemoptysis, and are often confused with lung cancer or tuberculosis. Paragonimiasis can be efficiently treated with praziquantel, but diagnosis is often delayed, and patients are frequently treated for other conditions. To improve diagnosis, we selected five Paragonimus kellicotti proteins based on transcriptional abundance, recognition by patient sera, and conservation among trematodes and expressed them as His-fusion proteins in Escherichia coli. Sequences for these proteins have 76-99% identity with amino acid sequences for orthologs in the genomes of Paragonimus westermani, Paragonimus heterotremus, and Paragonimus miyazakii. Immunohistology studies showed that antibodies raised to four recombinant proteins bound to the tegument of adult P. kellicotti worms, at the parasite host interface. Only a known egg antigen was absent from the tegument but present in developing and mature eggs. We evaluated the diagnostic potential of these antigens by Western blot with sera from patients with paragonimiasis (from MO and the Philippines), fascioliasis, and schistosomiasis, and with sera from healthy North American controls. Two recombinant proteins (a cysteine protease and a myoglobin) showed the highest sensitivity and specificity as diagnostic antigens, and they detected antibodies in sera from paragonimiasis patients with early or mature infections. In contrast, antibodies to egg yolk ferritin appeared to be specific marker for patients with adult fluke infections that produce eggs. Our study has identified and localized antigens that are promising for serodiagnosis of human paragonimiasis.


Subject(s)
Antibodies, Helminth/blood , Antigens, Helminth/immunology , Paragonimiasis/diagnosis , Paragonimus/immunology , Praziquantel/therapeutic use , Adult , Animals , Anthelmintics , Antigens, Helminth/metabolism , Asia , Gerbillinae , Humans , Immunohistochemistry , Paragonimiasis/metabolism , Paragonimiasis/parasitology , Paragonimus westermani/immunology , Recombinant Proteins , Sensitivity and Specificity , Serologic Tests
2.
Korean J Intern Med ; 30(1): 56-61, 2015 Jan.
Article in English | MEDLINE | ID: mdl-25589836

ABSTRACT

BACKGROUND/AIMS: Pleuropulmonary paragonimiasis produces no specific symptoms or radiologic findings, allowing for the possibility of misdiagnosis. We evaluated the specific clinical and pleural fluid features of pleuropulmonary paragonimiasis masquerading as pleural tuberculosis. METHODS: We retrospectively analyzed the clinical and radiologic characteristics of 20 patients diagnosed with pleuropulmonary paragonimiasis between 2001 and 2011. RESULTS: In total, 17 patients presented with respiratory symptoms, including dyspnea (30%), hemoptysis (20%), cough (20%), and pleuritic chest pain (15%). Chest radiographs revealed intrapulmonary parenchymal lesions, including air-space consolidation (30%), nodular opacities (20%), cystic lesions (15%), ground-glass opacities (10%), and pneumothorax (5%). A pleural fluid examination revealed eosinophilia, low glucose levels, and high lactate dehydrogenase (LDH) levels in 87%, 76%, and 88% of the patients, respectively. These traits helped to distinguish pleuropulmonary paragonimiasis from other pleural diseases such as parapneumonic effusion, malignancy, and pleural tuberculosis. CONCLUSIONS: Pleuropulmonary paragonimiasis is often initially misdiagnosed as other pleural diseases. Therefore, it is important to establish the correct diagnosis. In patients with unexplained pleural effusion living in paragonimiasis-endemic areas, pleural fluid obtained by thoracentesis should be examined to distinguish pleuropulmonary paragonimiasis. When marked eosinophilia, high LDH levels, and low glucose levels are identified in pleural fluid, physicians could consider a diagnosis of pleuropulmonary paragonimiasis.


Subject(s)
Lung Diseases, Parasitic/diagnosis , Paragonimiasis/diagnosis , Paragonimus westermani/isolation & purification , Pleural Effusion/diagnosis , Tuberculosis, Pleural/diagnosis , Adolescent , Adult , Aged , Animals , Biomarkers/analysis , Child , Child, Preschool , Diagnosis, Differential , Enzyme-Linked Immunosorbent Assay , Eosinophilia/diagnosis , Eosinophilia/parasitology , Female , Glucose/analysis , Humans , L-Lactate Dehydrogenase/analysis , Lung Diseases, Parasitic/diagnostic imaging , Lung Diseases, Parasitic/metabolism , Lung Diseases, Parasitic/parasitology , Male , Middle Aged , Paracentesis , Paragonimiasis/diagnostic imaging , Paragonimiasis/metabolism , Paragonimiasis/parasitology , Pleural Effusion/diagnostic imaging , Pleural Effusion/metabolism , Pleural Effusion/parasitology , Predictive Value of Tests , Retrospective Studies , Tomography, X-Ray Computed , Young Adult
3.
Int J Parasitol ; 40(14): 1651-8, 2010 Dec.
Article in English | MEDLINE | ID: mdl-20716443

ABSTRACT

We previously showed that adult Paragonimus westermani, the causative agent of paragonimiasis and whose habitat is the host lung, possesses both aerobic and anaerobic respiratory chains, i.e., cyanide-sensitive succinate oxidase and NADH-fumarate reductase systems, in isolated mitochondria (Takamiya et al., 1994). This finding raises the intriguing question as to whether adult Paragonimus worms possess two different populations of mitochondria, one having an aerobic succinate oxidase system and the other an anaerobic fumarate reductase system, or whether the worms possess a single population of mitochondria possessing both respiratory chains (i.e., mixed-functional mitochondria). Staining of trematode tissues for cytochrome c oxidase activity showed three types of mitochondrial populations: small, strongly stained mitochondria with many cristae, localised in the tegument and tegumental cells; and two larger parenchymal cell mitochondria, one with developed cristae and the other with few cristae. The tegumental and parenchymal mitochondria could be separated by isopycnic density-gradient centrifugation and showed different morphological characteristics and respiratory activities, with low-density tegumental mitochondria having cytochrome c oxidase activity and high-density parenchymal mitochondria having fumarate reductase activity. These results indicate that Paragonimus worms possess three different populations of mitochondria, which are distributed throughout trematode tissues and function facultatively, rather than having mixed-functional mitochondria.


Subject(s)
Dog Diseases/parasitology , Mitochondria/metabolism , Oxygen/metabolism , Paragonimiasis/veterinary , Paragonimus westermani/physiology , Adaptation, Physiological , Aerobiosis , Anaerobiosis , Animals , Dog Diseases/metabolism , Dogs , Electron Transport Complex IV/metabolism , Helminth Proteins , Host-Parasite Interactions , Lung/metabolism , Lung/parasitology , Mitochondria/enzymology , Oxidoreductases Acting on CH-CH Group Donors/metabolism , Paragonimiasis/metabolism , Paragonimiasis/parasitology , Paragonimus westermani/enzymology
4.
Neurosci Res ; 65(2): 141-7, 2009 Oct.
Article in English | MEDLINE | ID: mdl-19539668

ABSTRACT

Excretory-secretory products (ESP) from helminthic parasites may play pivotal roles in the immune regulation in hosts. Previously, we reported that ESP produced from Paragonimus westermani induced morphological activation of microglial cells and markedly stimulated nitric oxide (NO) production via activation of mitogen-activated protein kinases (MAPKs). In the present study, we investigated the role of protein kinase C and protein kinase A in MAPKs-dependent NO production by ESP. We found that treatment with protein kinase C inhibitor Go6976 strongly inhibited the phosphorylation of p38 and JNK, but not ERK, of MAPKs and decreased the production of NO in ESP-stimulated microglial cells. Inhibition of ERK, p38 or PKC decreased the ESP-induced activation of NF-kappaB, an important transcription factor for iNOS expression. Furthermore, ESP increased the level of p-CREB in microglial cells. However, adenylyl cyclase activator (forskolin), adenylyl cyclase inhibitor (SQ22536), cAMP analogue (db-cAMP) or protein kinase A inhibitor (H89) was not able to change iNOS expression and NO production in ESP-treated microglial cells. It implies that the cAMP-PKA-CREB pathway is not implicated in the ESP-evoked NO production in microglial cells. Thus, our results indicate that ESP stimulates microglial expression of iNOS via both PKC-dependent and -independent MAPKs phosphorylation and NF-kappaB activation.


Subject(s)
Encephalitis/metabolism , Microglia/metabolism , Nitric Oxide/metabolism , Paragonimiasis/metabolism , Paragonimus westermani/metabolism , Protein Kinase C/metabolism , Animals , Brain/metabolism , Brain/parasitology , Brain/physiopathology , Cells, Cultured , Central Nervous System Infections/metabolism , Central Nervous System Infections/parasitology , Central Nervous System Infections/physiopathology , Cyclic AMP/metabolism , Cyclic AMP Response Element-Binding Protein/metabolism , Cyclic AMP-Dependent Protein Kinases/metabolism , Encephalitis/parasitology , Encephalitis/physiopathology , Enzyme Inhibitors/pharmacology , JNK Mitogen-Activated Protein Kinases/metabolism , MAP Kinase Signaling System/physiology , Microglia/parasitology , NF-kappa B/metabolism , Nitric Oxide Synthase Type II/metabolism , Paragonimiasis/physiopathology , Phosphorylation/drug effects , Phosphorylation/physiology , Rats , Rats, Sprague-Dawley , p38 Mitogen-Activated Protein Kinases/metabolism
5.
Korean J Parasitol ; 46(2): 95-9, 2008 Jun.
Article in English | MEDLINE | ID: mdl-18552546

ABSTRACT

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.


Subject(s)
Cell Degranulation , Cysteine Endopeptidases/immunology , Eosinophils/immunology , Helminth Proteins/immunology , Paragonimiasis/immunology , Paragonimus westermani/enzymology , Superoxides/immunology , Animals , Astacoidea/parasitology , Cysteine Endopeptidases/isolation & purification , Eosinophils/metabolism , Helminth Proteins/isolation & purification , Humans , Paragonimiasis/metabolism , Paragonimus westermani/isolation & purification
6.
Proteomics ; 6(4): 1290-300, 2006 Feb.
Article in English | MEDLINE | ID: mdl-16404718

ABSTRACT

Paragonimus westermani causes inflammatory lung disease in humans. The parasite excretes a host of biologically active molecules, which are thought to be involved in pathophysiological and immunological events during infection. Analyses of the 2-DE protein profiles of the excretory-secretory products (ESP) of adult P. westermani revealed approximately 147 protein spots, at least 15 of which were identified as cysteine proteases (CPs), at pHs between 4.5 and 8.5, and molecular weights (MWs) between 27 and 35 kDa. An additional three CPs (designated as PwCP-3, -8 and -11) were newly recognized by TOF/TOF MS. Their molecular biological information, which shared a high level sequence homology, was elucidated. The majority of the CPs reacted strongly with sera from paragonimiasis patients. When we observed the chronological changes in the antibody responses of the respective CPs against canine sera collected serially at 1, 3, 5, 7, 11 and 14 wk after experimental infection, these molecules exhibited a multiplicity of distinct immune recognition patterns. Our results clearly showed that P. westermani adult ESP were principally composed of excretory-secretory CPs, and that these CPs may exert effects not only on host tissue degradation and nutrient uptake, but also on the immune-regulating cells via synergistic and independent interactions.


Subject(s)
Cysteine Endopeptidases , Helminth Proteins/immunology , Paragonimiasis/immunology , Paragonimus westermani/enzymology , Proteome/analysis , Amino Acid Sequence , Animals , Biomarkers/metabolism , Cysteine Endopeptidases/immunology , Cysteine Endopeptidases/metabolism , Dogs , Electrophoresis, Gel, Two-Dimensional , Helminth Proteins/metabolism , Humans , Molecular Sequence Data , Paragonimiasis/diagnosis , Paragonimiasis/metabolism , Paragonimus westermani/growth & development , Paragonimus westermani/immunology , Sequence Homology, Amino Acid , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
7.
Clin Exp Immunol ; 123(1): 94-8, 2001 Jan.
Article in English | MEDLINE | ID: mdl-11168004

ABSTRACT

To investigate the pathogenic mechanisms of eosinophilic pleural effusion in patients with paragonimiasis, we measured the levels of IL-5, granulocyte-macrophage colony-stimulating factor (GM-CSF) and interferon-gamma (IFN-gamma) in pleural effusions. Samples were obtained from 11 patients with Paragonimus westermani infection. In addition, samples from 12 patients with pleural transudates, 16 with tuberculous pleurisy, seven with empyema and 20 with lung cancer were also examined. Eosinophilia was remarkable in peripheral blood (range 4-34%, median 23.4%) and pleural fluid (range 0-95%, median 71%) of paragonimiasis patients. IL-5 concentrations in pleural effusions of paragonimiasis were markedly higher than those in other groups. Although marked elevation of GM-CSF and IFN-gamma levels was observed in pleural effusion of empyema and tuberculosis patients, it was marginal in the pleural effusion of paragonimiasis patients. In paragonimiasis patients, IL-5 levels in the pleural effusion correlated well with the percentage of eosinophils in peripheral blood and pleural fluid. Such a correlation was not observed between GM-CSF levels in pleural effusion and percentages of eosinophils in pleural fluid or peripheral blood. Our findings suggest that in paragonimiasis IL-5 in the local inflammatory site is particularly important in mediating eosinophilia in peripheral blood and pleural effusion.


Subject(s)
Interleukin-5/metabolism , Paragonimiasis/immunology , Pleural Effusion/immunology , Pleural Effusion/metabolism , Adult , Aged , Cell Movement/immunology , Eosinophilia/blood , Eosinophilia/immunology , Eosinophilia/parasitology , Female , Granulocyte-Macrophage Colony-Stimulating Factor/metabolism , Humans , Interferon-gamma/metabolism , Interleukin-5/blood , Leukocyte Count , Male , Middle Aged , Paragonimiasis/blood , Paragonimiasis/metabolism , Pleural Effusion/parasitology , Pleural Effusion/pathology
8.
Intern Med ; 39(5): 433-6, 2000 May.
Article in English | MEDLINE | ID: mdl-10830191

ABSTRACT

We report a case of Paragonimus westermani infection with a reticulonodular lesion in the right lung, left pleural effusion, and a mobile subcutaneous mass. Analyses of pleural effusion and bronchoalveolar lavage fluid (BALF) showed marked eosinophilia and high levels of eosinophil cationic protein and interleukin (IL)-5. Transbronchial lung biopsy revealed the presence of pneumonia with mild eosinophilic infiltration but remarkable lymphocytic infiltration. In this patient, high IL-5 levels in both BALF and pleural effusion could explain the remarkable eosinophilia.


Subject(s)
Lung Diseases, Parasitic/diagnosis , Paragonimiasis/diagnosis , Ribonucleases , Skin Diseases, Parasitic/diagnosis , Aged , Blood Proteins/metabolism , Bronchoalveolar Lavage Fluid/chemistry , Eosinophil Granule Proteins , Eosinophilia/etiology , Female , Humans , Interleukin-5/metabolism , Lung Diseases, Interstitial/etiology , Paragonimiasis/complications , Paragonimiasis/metabolism , Pleural Effusion/metabolism
9.
Am J Trop Med Hyg ; 26(4): 688-92, 1977 Jul.
Article in English | MEDLINE | ID: mdl-889010

ABSTRACT

Menichlopholan, biphenyl compound, in a single dose of 2 mg/kg body weight, gave a 73%-90% cure rate in the treatment of pulmonary infections due to Paragonimus uterobilateralis at two centers in Nigeria. Ninety-five patients were followed up for 4 months, and 58 for 1 year. Side effects of the drug included sweating and body pains, but there was no significnat biochemical or hematological evidence of damage to the internal organs. The advantages of single dose therapy are likely to make menichlopholan the treatment of choice for paragonimiasis in Africa, and probably elsewhere.


Subject(s)
Lung Diseases, Parasitic/drug therapy , Niclofolan/therapeutic use , Nitrophenols/therapeutic use , Paragonimiasis/drug therapy , Adolescent , Adult , Bithionol/therapeutic use , Child , Female , Hemoptysis/etiology , Humans , Liver Function Tests , Lung/diagnostic imaging , Lung Diseases, Parasitic/complications , Lung Diseases, Parasitic/diagnostic imaging , Lung Diseases, Parasitic/metabolism , Male , Middle Aged , Niclofolan/adverse effects , Paragonimiasis/complications , Paragonimiasis/diagnostic imaging , Paragonimiasis/metabolism , Parasite Egg Count , Radiography
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