Backgrounds: After publishing a paper shown diarrheagenic and emetic activity with Kudoa septempuctata spores inoculated on 4-5-day old ddY mice, the Centers for Disease Control and Prevention in Korea reported 11 events occurred in 2015 as ‘Kudoa food posioning’. Because epidemiological design of the report was not an analytical but a descriptive study, it needs to evaluate evidences related to pathogenicity of K. septempuctata. Methods: Related articles showing evidences of pathogenicity were searched using citation discovery tool in PubMed based on the first report worldwide. Information for kinds of experimental animal, oral administrated spores and the results were extracted. Results: Four papers evaluating pathogenicity of Kudoa species were selected. Among them, the first-reported paper only suggested the pathogenicity. Other 3 papers reported no abnormal symptoms and histo-pathologic changes. Conclusions: Based on these controversies, the evidences for pathogenicity of K. septempuctata are weak and make a suspicion about pathogenic suggestion. Thus further studies evaluating pathogenicity of K. septempuctata are need urgently. Keyword: Fish parasitology, Virulence, Food parasitology, Parasitic intestinal diseases, Myxozoa

After publishing results of a study that revealed diarrheagenic and emetic activity in 4-5-day old mice infected with Kudoa septempunctata (K. septempunctata) spores, the Korea Centers for Disease Control and Prevention reported 11 events of "Kudoa food poisoning" in 2015. The epidemiological design of the previous study was descriptive rather than analytical; therefore, this study aimed to further investigate the pathogenicity of K. septempunctata. Academic articles showing evidence of the pathogenicity of K. septempunctata were searched via PubMed using the citation discovery tool. Information regarding the kinds of experimental animals and inoculum spores used, as well as study results were extracted. Four articles evaluating the pathogenicity of Myxospran parasites were selected; the first article suggested the pathogenicity of K. septempunctata, while the remaining three articles reported no abnormal symptoms or histopathologic changes. Our findings indicate that there is weak evidence supporting the pathogenicity of K. septempunctata. Further studies evaluating the pathogenicity of K. septempunctata are needed urgently.

A case of Taenia asiatica infection detected by small bowel series and colonoscopy is described. The patient was a 42-year-old Korean man accompanied by discharge of movable proglottids via anus. He used to eat raw pig liver but seldom ate beef. Small bowel series radiologic examinations showed flat tape-like filling defects on the ileum. By colonoscopy, a moving flat tapeworm was observed from the terminal ileum to the ascending colon. The tapeworm was identified as T. asiatica by mitochondrial DNA sequencing. The patient was prescribed with a single oral dose (16 mg/kg) of praziquantel.

PURPOSE:Recent studies have used the term "gastroallergic anisakiasis" to describe incidental gastrointestinal infection with Anisakis spp. larvae, proposed as a causative agent of food hypersensitivity. However, it is unknown whether this condition represents an independent disease entity distinguishable from acute gastric anisakiasis. To better understand the role of the allergic response in Anisakis infections we examined the clinical and immunological implications of Anisakis-specific IgE. METHODS:A prospective study was performed in a geographic region where the consumption of raw seafood is common. Case subjects who had been clinically diagnosed with gastroallergic anisakiasis were selected, along with controls who frequently ate raw seafood but had never experienced gastroallergic anisakiasis-like symptoms. Clinical and immunological features were compared based on atopic status, sensitization rates to Anisakis, and serum titer of Anisakis-specific IgE. RESULTS:Seventeen case subjects and 135 controls were included in this study. The case subjects had experienced gastrointestinal symptoms after raw seafood ingestion, along with additional mucocutaneous, respiratory, or multisystemic symptoms. Case subjects were significantly sensitized to Anisakis excretory-secretory product and crude extract compared with controls (76.5% vs 19.3%, P<0.001, and 88.2% vs 30.3%, P<0.001, respectively). Anisakis-specific serum IgE titers were also significantly higher in case subjects than in controls. Both the results of skin prick tests and elevated Anisakis-specific IgE titers (>17.5 kU/L) were found to be reliable indicators for the diagnosis of gastroallergic anisakiasis. CONCLUSIONS:Among patients presenting acute gastric anisakiasis-like symptoms, a diagnosis of gastroallergic anisakiasis may be strongly supported by a high Anisakis-specific IgE titer.

The authors partially purified glutathione S-transferase (GST) of Neodiplostomum seoulense monitoring its activity using 1-chloro-2,4-dinitrobenzene (CDNB). The enzyme was partially purified using glutathione?agarose affinity columnchromatography. The purification fold and recovery rate were 9.7 and 50.4%, respectively. The molecular weight of thepurified GST was estimated to be 28 kDa on 7.5-15% gradient SDS-PAGE. From this study, the authors firstly reported thepresence of GST from N. seoulense and further studies such as inhibition test, subclass of GST as well as immunologicalstudies would be required.

Panonychus citri damages the leaves of citrus trees, causing defoliation, and induces T-helper type 2 (T H 2) immune responses (occupational asthma) via a hitherto unknown mechanism. This is a particular problem on Jeju Island, which is located to the south of the Korean peninsula. In this study, we show for the first time how P. citri induces TH2 immunity. Exposure to P. citri induces the production of thymic stromal lymphopoietin (TSLP) by either basophils or CD4+ T cells (it is not certain which), which results in the production of interleukin 4 (IL-4). IL-4 promotes the production of immunoglobulin E (IgE), which ultimately contributes to the process of allergic inflammation. Therefore, TSLP plays an important role in the P. citri-induced T H 2 immune response.

Several studies have reported that the citrus red mites Panonychus citri were an important allergen of citrus-cultivating farmers in Jeju Island. The aim of the present study was to purify and assess properties of a cysteine protease from the mites acting as a potentially pathogenic factor to citrus-cultivating farmers. A cysteine protease was purified using column chromatography of Mono Q anion exchanger and Superdex 200 HR gel filtration. It was estimated to be 46 kDa by gel filtration column chromatography and consisted of 2 polypeptides, at least. Cysteine protease inhibitors, such as trans poxy-succinyl-L-leucyl-amido (4-guanidino) butane (E-64) and iodoacetic acid (IAA) totally inhibited the enzyme activities, whereas serine or metalloprotease inhibitors did not affect the activities. In addition, the purified enzyme degraded human IgG, collagen, and fibronectin, but not egg albumin. From these results, the cysteine protease of the mites might be involved in the pathogenesis such as tissue destruction and penetration instead of nutrient digestion.

PURPOSE:To evaluate the clinical and CT findings in patients with small-bowel Anisakiasis.MATERIALS AND METHODS:Nineteen patients with small-bowel Anisakiasis and who underwent abdominal CT between 2005 and 2012 were enrolled in our study. All of these patients were diagnosed using either a serologic test for Anisakiasis (n = 18) or by pathology (n = 1). Their medical records were reviewed in order to determine the clinical findings. CT images were retrospectively reviewed by two radiologists to evaluate the characteristics of the involved bowel wall and the ancillary findings.RESULTS:All patients had presented with the acute onset of severe abdominal pain as well as a history of having recently eaten raw fish. The mean time interval from eating the fish to the onset of abdominal pain was 1.7 days. Eighteen patients were treated conservatively and experienced resolution of their symptoms within seven days of hospitalization. One patient underwent surgical exploration for presumed small-bowel ischemia. The sites of involvement included the ileum (16/19, 84%) and jejunum (3/19, 16%). All patients had circumferential bowel-wall thickening (mean, 0.8 cm) with an intermediate length of involved bowel (mean, 7.9 cm). Small-bowel obstruction occurred in 16 patients (84%). The target sign was present in 17 patients (89%), ascites, particularly in the perilesional area, in 16 patients (84%), and mesenteric edema in 15 patients (79%).CONCLUSION:Small-bowel Anisakiasis should be considered in the differential diagnosis of acute abdomen in order to avoid unnecessary surgery when patients present with abdominal pain after having recently eaten raw fish, concentric bowel-wall thickening with the target sign in the ileum, perilesional ascites, as well as bowel obstruction seen on CT.

A cysteine protease from excretion-secretion products of adult Philippine Fasciola gigantica was purified to homogeneity by sequential gel filtration and anion exchange chromatographies, FPLC Resource Q anion exchange chromatography and then characterized biochemically. The molecular weight was estimated at 26 kDa by SDS-PAGE. It exhibited marked increase in proteolytic activity in the presence of dithiotreitol, slight activity with phenylmethyl sulphonyl fluoride but was totally inhibited by cysteine protease inhibitors, iodoacetic acid and leupeptin. Optimal activity was observed at pH 4 and at temperature of 35oC. It was stable at pH 4.5 to 5 when stored at 4oC. The enzyme was able to hydrolyze bovine serum albumin and bovine immunoglobulin G but not gelatin and hemoglobin at acidic pH. In addition, it was antigenic. This cysteine protease from adult Philippine F. gigantica may have potential use in diagnosis and prevention of ruminant fasciolosis. Keywords: characterization, cysteine protease, Fasciola gigantica, fasciolosis, purification

The evaluation of sensitization to aeroallergens is valuable in diagnosis and treatment of atopic respiratory disorders. Because of the identical climates in Jeju, Korea, the sensitization rates may differ from other regions of Korea. The sensitization rates in Jeju was rarely reported. To examine the sensitization rates to common inhalant allergens and to suggest the proper evaluation panels of skin prick tests in Jeju, Korea The results of skin prick tests in atopic individuals were analyzed retrospectively and cross-sectionally from allergy clinics. Two hundred and eighty seven atopic individuals (mean age of 33) were enrolled from 3regional facilities. Skin prick tests were performed with different allergens from different manufacturers. House dust mites, cockroach, oak and mugwort are commonly sensitized aeroal1ergens in Jeju. Some of allergens showed different sensitization rates according to the manufacturers. Common aeroallergens for skin prick tests in Jeju were suggested. Standardization of aeroallergens for skin prick tests should be performed.

Hypersensitivity to human seminal fluid is rare but can be life threatening. We report a case of IgE-mediated anaphylaxis to seminal plasma that was diagnosed by skin prick tests and successfully treated by local desensitization. A 32-year-old woman suffering from angioedema and hypotension after exposure to semen was treated with epinephrine upon admission. Skin prick tests and immunoblotting for IgE binding components showed that she was sensitized to her husband's seminal plasma. Local desensitization, which persisted for six months, was achieved by intravaginal administration of serial dilutions of her husband's seminal plasma.

Helminthic cysteine proteases are well known to play critical roles in tissue invasion, nutrient uptake, and immune evasion of the parasites. In the same manner, the sparganum, the plerocercoid of Spirometra mansoni, is also known to secrete a large amount of cysteine proteases. However, cysteine protease inhibitors regulating the proteolytic activities of the cysteine protease are poorly illustrated. In this regard, we partially purified an endogenous cysteine protease inhibitor from spargana and characterized its biochemical properties. The cysteine protease inhibitor was purified by sequential chromatographies using Resource Q anion exchanger and Superdex 200 HR gel filtration from crude extracts of spargana. The molecular weight of the purified protein was estimated to be about 11 kD on SDS-PAGE. It was able to inhibit papain and 27 1 cysteine protease of spargana with the ratio of 25.7% and 49.1%, respectively, while did not inhibit chymotrypsin. This finding suggests that the cysteine protease inhibitor of spargana may be involved in regulation of endogenous cysteine proteases of the parasite, rather than interact with cysteine proteases from their hosts.

The aim of this study is to determine the Toxocara seropositive rate among healthy people with eosinophilia. A total of 97 people residing in Seoul who were healthy and whose blood eosinophilia was over 10%, as shown by regular health check-ups in 2004, were subjected to this study. Their sera were tested by immunoblotting and ELISA with the antigen of larval Toxocara canis excretory-secretory (ES) protein. Sixty-five sera were band-positive (67.0%). The seropositve control sera were positive to band sizes of 66 kDa, 56 kDa, 32 kDa, and 13 kDa. In ELISA, 63 sera (65.0%) were positive to T. canis ES protein. There was no significant correlation between the IgG ELISA titer and the level of eosinophilia (r = 0.156, P = 0.156). As there were insufficient data to determine whether there were cross-reactions with other helminthic infections, or whether atopy occurred, further studies are required to verify the cause of the seropositive reactions against T. canis ES antigen. Toxocariasis seropositivity is suggested to be the major cause of eosinophilia, since the Toxocara seroprevalence among Korean rural adults was shown to be approximately 5%.

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.

A cysteine protease activity of Panonychus citri was detected and partially characterized. Proteolytic activity of the enzyme was enhanced by the addition of reducing agent, dithiothreitol (DTT), and also it was absolutely inhibited by cysteine protease specific inhibitors, such as trans-epoxy-succinyl-L-leucyl-amido (4-guanidino) butane (E.64) and iodoacetic acid (IAA). The other specific inhibitors of serine- and metallo-proteases could not inhibit the enzyme activity. This result showed for the first time cysteine protease activity of P. citri. Further studies on the biological roles of the enzyme are required.

A 29 kDa cysteine protease of Taenia solium metacestodes was purified by Mono Q anion-exchanger and Superose 6 HR gel filtration chromatography. The enzyme was effectively inhibited by cysteine protease inhibitors, such as iodoacetic acid (IAA) and trans-epoxy-succinyl-L-leucyl-amido (4-guanidino) butane (E-64) while inhibitors acting on serine- or metallo-proteases did not affect the enzyme activity. The purified enzyme degraded human immunoglobulin G (IgG), collagen and bovine serum albumin (BSA), but human IgG was more susceptible for proteolysis by the enzyme. To define the precise biological roles of the enzyme, more detailed biochemical and functional studies would be required.

Eosinophil degranulation is considered to be a key effector function for the killing of helminthic worms and tissue inflammation at worm-infected lesion sites. However, relatively little data are available with regard to eosinophil response after stimulation with worm-secreted products which contain a large quantity of cysteine proteases. In this study, we attempted to determine whether the degranulation of human eosinophils could be induced by the direct stimulation of the excretory-secretory products (ESP) of Paragonimus westermani, which causes pulmonary paragonimiasis in human beings. Incubation of eosinophils for 3 hr with Paragonimus-secreted products resulted in marked degranulation, as evidenced by the release of eosinophil-derived neurotoxin (EDN) in the culture supernatants. Moreover, superoxide anion was produced by eosinophils after stimulation of the ESP. The ESP-induced EDN release was found to be significantly inhibited when the ESP was pretreated with protease inhibitor cocktail or the cysteine protease inhibitor, E-64. These findings suggest that human eosinophils become degranulated in response to P. westermani-secreted proteases, which may contribute to in vivo tissue inflammation around the worms.

A highly specific antigenic protein of 31 kDa from plerocercoid of Spirometra mansoni (sparganum) was obtained by gelatin affinity and Mono Q anion-exchange column chromatography. The purified 31 kDa protein was subjected to N-glycan enzymatic digestion for structural analysis. The relative electrophoretic mobility was analyzed by SDS-PAGE, before and after digestion. On SDS-PAGE after enzymatic digestion, the 31 kDa protein showed a molecular shift of approximately 2 kDa, which indicated the possession of complex N-linked oligosaccharides (N-glycosidase F sensitive) but not of high-mannose oligosaccharides (endo-beta-N-acetylglucosaminidase H, non-sensitive). Chemically periodated 31 kDa protein showed statistically non-significant changes with human sparganosis sera by enzyme linked immunosorbent assay (ELISA). Therefore, the dominant epitopes of the 31 kDa molecule in human sparganosis were found to be mainly polypeptide, while N-glycans of the antigenic molecule in sparganum was minimal in anti-carbohydrate antibody production.

The 150 kDa protein of cyst fluid (CF) of Taenia solium metacestodes was purified by ammonium sulfate fractionation and Superose 6 HR gel filtration chromatography. The purified protein consisted of three subunits (15, 10 and 7 kDa proteins), which were analyzed with the use of a 7.5-15% gradient sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Immunofluorescence study was carried out by using immunize specific polyclonal antibody. Positive reactions were noticed at bladder walls, calcareous corpuscles, granules of cyst fluid and some host tissue surrounding the bladder wall of the metacestodes. These results suggest that the 150 kDa protein was secreted into host tissues, inducing immune responses in the host, and it may play important roles in the cellular physiology of the parasites.

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