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Zhong Quan Wang - One of the best experts on this subject based on the ideXlab platform.
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Molecular cloning and characterization of a Spirometra erinacei casein kinase I.
Parasitology Research, 2015Co-Authors: Han Wang, Ming Guo Xu, Li Fu Wang, Xin Qi, Man Wang, Xi Zhang, Peng Jiang, Zhong Quan WangAbstract:The Spirometra erinacei casein kinase I (SeCKI) gene was cloned and expressed in Escherichia coli, and its characteristics were investigated in this study. The recombinant SeCP protein (rSeCKI) was purified. The vaccination of mice with rSeCKI induced the Th1/Th2-mixed type of immune response with Th2 predominant (high levels of IgG1). Western blotting analysis showed that rSeCP was recognized by the sera of Plerocercoid-infected mice, and anti-rSeCP serum recognized the native SeCP protein of Plerocercoid crude antigens. Transcription and expression of SeCP was observed at the Plerocercoid and adult stages of S. erinacei. Immunolocalization identified SeCKI in the tegument and parenchymal tissues of Plerocercoids and in the teguments of adults. SeCKI appeared to be essential indispensable for the S. erinacei development and survival in host, but its biological functions need to be further investigated.
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characterization of spirometra erinaceieuropaei Plerocercoid cysteine protease and potential application for serodiagnosis of sparganosis
PLOS Neglected Tropical Diseases, 2015Co-Authors: Li Na Liu, Zhong Quan Wang, Xi Zhang, Peng Jiang, Ruo Dan Liu, Zi Fang Zhang, Jing CuiAbstract:Background Sparganosis is a neglected but important food-borne parasitic zoonosis. Clinical diagnosis of sparganosis is difficult because there are no specific manifestations. ELISA using Plerocercoid crude or excretory–secretory (ES) antigens has high sensitivity but has cross-reactions with other helminthiases. The aim of this study was to characterize Spirometra erinaceieuropaei cysteine protease (SeCP) and to evaluate its potential application for serodiagnosis of sparganosis. Methodology/Principal Findings The full length SeCP gene was cloned, and recombinant SeCP (rSeCP) was expressed and purified. Western blotting showed that rSeCP was recognized by the serum of sparganum-infected mice, and anti-rSeCP serum recognized the native SeCP protein of Plerocercoid crude or ES antigens. Expression of SeCP was observed at Plerocercoid stages but not at the adult and egg stages. Immunolocalization identified SeCP in Plerocercoid tegument and parenchymal tissue. The rSeCP had CP activity, and the optimum pH and temperature were 5.5 and 37°C, respectively. Enzymatic activity was significantly inhibited by E-64. rSeCP functions to degrade different proteins and the function was inhibited by anti-rSeCP serum and E-64. Immunization of mice with rSeCP induced Th2-predominant immune responses and anti-rSeCP antibodies had the potential capabilities to kill Plerocercoids in an ADCC assay. The sensitivity of rSeCP-ELISA and ES antigen ELISA was 100% when performed on sera of patients with sparganosis. The specificity of rSeCP-ELISA and ES antigen ELISA was 98.22% (166/169) and 87.57% (148/169), respectively (P<0.05). Conclusions The rSeCP had the CP enzymatic activity and SeCP seems to be important for the survival of Plerocercoids in host. The rSeCP is a potential diagnostic antigen for sparganosis.
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Expression and immunolocalization of SeCP at different developmental stages of S. erinaceieuropaei.
2015Co-Authors: Li Na Liu, Zhong Quan Wang, Xi Zhang, Peng Jiang, Ruo Dan Liu, Zi Fang Zhang, Jing CuiAbstract:The results of IFT with the sections of S. erinaceieuropaei Plerocercoids and adult worms reacted with anti-rSeCP serum. The immunostaining is seen at the teguments and parenchymal tissues of Plerocercoids from mice (A) and Plerocercoids from frogs (B), but no immunostaining was observed in the sections of gravid and eggs (C), mature (D), immature (E) proglottids, and scolex and neck (F) of adult worm. The Plerocercoid reacted with serum of mice infected with Plerocercoids as a positive control (G), and did not show recognition by normal mouse serum (H) and PBS (I) as a negative control.
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Characterization of Spirometra erinaceieuropaei Plerocercoid Cysteine Protease and Potential Application for Serodiagnosis of Sparganosis
2015Co-Authors: Li Na Liu, Zhong Quan Wang, Xi Zhang, Peng Jiang, Ruo Dan Liu, Zi Fang Zhang, Jing CuiAbstract:BackgroundSparganosis is a neglected but important food-borne parasitic zoonosis. Clinical diagnosis of sparganosis is difficult because there are no specific manifestations. ELISA using Plerocercoid crude or excretory–secretory (ES) antigens has high sensitivity but has cross-reactions with other helminthiases. The aim of this study was to characterize Spirometra erinaceieuropaei cysteine protease (SeCP) and to evaluate its potential application for serodiagnosis of sparganosis.Methodology/Principal FindingsThe full length SeCP gene was cloned, and recombinant SeCP (rSeCP) was expressed and purified. Western blotting showed that rSeCP was recognized by the serum of sparganum-infected mice, and anti-rSeCP serum recognized the native SeCP protein of Plerocercoid crude or ES antigens. Expression of SeCP was observed at Plerocercoid stages but not at the adult and egg stages. Immunolocalization identified SeCP in Plerocercoid tegument and parenchymal tissue. The rSeCP had CP activity, and the optimum pH and temperature were 5.5 and 37°C, respectively. Enzymatic activity was significantly inhibited by E-64. rSeCP functions to degrade different proteins and the function was inhibited by anti-rSeCP serum and E-64. Immunization of mice with rSeCP induced Th2-predominant immune responses and anti-rSeCP antibodies had the potential capabilities to kill Plerocercoids in an ADCC assay. The sensitivity of rSeCP-ELISA and ES antigen ELISA was 100% when performed on sera of patients with sparganosis. The specificity of rSeCP-ELISA and ES antigen ELISA was 98.22% (166/169) and 87.57% (148/169), respectively (P
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Detection of anti-Plerocercoid IgG antibodies in serum samples of patients with sparganosis and other parasitosis by rSeCP-ELISA and ES-ELISA.
2015Co-Authors: Li Na Liu, Zhong Quan Wang, Xi Zhang, Peng Jiang, Ruo Dan Liu, Zi Fang Zhang, Jing CuiAbstract:Detection of anti-Plerocercoid IgG antibodies in serum samples of patients with sparganosis and other parasitosis by rSeCP-ELISA and ES-ELISA.
Jing Cui - One of the best experts on this subject based on the ideXlab platform.
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characterization of spirometra erinaceieuropaei Plerocercoid cysteine protease and potential application for serodiagnosis of sparganosis
PLOS Neglected Tropical Diseases, 2015Co-Authors: Li Na Liu, Zhong Quan Wang, Xi Zhang, Peng Jiang, Ruo Dan Liu, Zi Fang Zhang, Jing CuiAbstract:Background Sparganosis is a neglected but important food-borne parasitic zoonosis. Clinical diagnosis of sparganosis is difficult because there are no specific manifestations. ELISA using Plerocercoid crude or excretory–secretory (ES) antigens has high sensitivity but has cross-reactions with other helminthiases. The aim of this study was to characterize Spirometra erinaceieuropaei cysteine protease (SeCP) and to evaluate its potential application for serodiagnosis of sparganosis. Methodology/Principal Findings The full length SeCP gene was cloned, and recombinant SeCP (rSeCP) was expressed and purified. Western blotting showed that rSeCP was recognized by the serum of sparganum-infected mice, and anti-rSeCP serum recognized the native SeCP protein of Plerocercoid crude or ES antigens. Expression of SeCP was observed at Plerocercoid stages but not at the adult and egg stages. Immunolocalization identified SeCP in Plerocercoid tegument and parenchymal tissue. The rSeCP had CP activity, and the optimum pH and temperature were 5.5 and 37°C, respectively. Enzymatic activity was significantly inhibited by E-64. rSeCP functions to degrade different proteins and the function was inhibited by anti-rSeCP serum and E-64. Immunization of mice with rSeCP induced Th2-predominant immune responses and anti-rSeCP antibodies had the potential capabilities to kill Plerocercoids in an ADCC assay. The sensitivity of rSeCP-ELISA and ES antigen ELISA was 100% when performed on sera of patients with sparganosis. The specificity of rSeCP-ELISA and ES antigen ELISA was 98.22% (166/169) and 87.57% (148/169), respectively (P<0.05). Conclusions The rSeCP had the CP enzymatic activity and SeCP seems to be important for the survival of Plerocercoids in host. The rSeCP is a potential diagnostic antigen for sparganosis.
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Expression and immunolocalization of SeCP at different developmental stages of S. erinaceieuropaei.
2015Co-Authors: Li Na Liu, Zhong Quan Wang, Xi Zhang, Peng Jiang, Ruo Dan Liu, Zi Fang Zhang, Jing CuiAbstract:The results of IFT with the sections of S. erinaceieuropaei Plerocercoids and adult worms reacted with anti-rSeCP serum. The immunostaining is seen at the teguments and parenchymal tissues of Plerocercoids from mice (A) and Plerocercoids from frogs (B), but no immunostaining was observed in the sections of gravid and eggs (C), mature (D), immature (E) proglottids, and scolex and neck (F) of adult worm. The Plerocercoid reacted with serum of mice infected with Plerocercoids as a positive control (G), and did not show recognition by normal mouse serum (H) and PBS (I) as a negative control.
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Characterization of Spirometra erinaceieuropaei Plerocercoid Cysteine Protease and Potential Application for Serodiagnosis of Sparganosis
2015Co-Authors: Li Na Liu, Zhong Quan Wang, Xi Zhang, Peng Jiang, Ruo Dan Liu, Zi Fang Zhang, Jing CuiAbstract:BackgroundSparganosis is a neglected but important food-borne parasitic zoonosis. Clinical diagnosis of sparganosis is difficult because there are no specific manifestations. ELISA using Plerocercoid crude or excretory–secretory (ES) antigens has high sensitivity but has cross-reactions with other helminthiases. The aim of this study was to characterize Spirometra erinaceieuropaei cysteine protease (SeCP) and to evaluate its potential application for serodiagnosis of sparganosis.Methodology/Principal FindingsThe full length SeCP gene was cloned, and recombinant SeCP (rSeCP) was expressed and purified. Western blotting showed that rSeCP was recognized by the serum of sparganum-infected mice, and anti-rSeCP serum recognized the native SeCP protein of Plerocercoid crude or ES antigens. Expression of SeCP was observed at Plerocercoid stages but not at the adult and egg stages. Immunolocalization identified SeCP in Plerocercoid tegument and parenchymal tissue. The rSeCP had CP activity, and the optimum pH and temperature were 5.5 and 37°C, respectively. Enzymatic activity was significantly inhibited by E-64. rSeCP functions to degrade different proteins and the function was inhibited by anti-rSeCP serum and E-64. Immunization of mice with rSeCP induced Th2-predominant immune responses and anti-rSeCP antibodies had the potential capabilities to kill Plerocercoids in an ADCC assay. The sensitivity of rSeCP-ELISA and ES antigen ELISA was 100% when performed on sera of patients with sparganosis. The specificity of rSeCP-ELISA and ES antigen ELISA was 98.22% (166/169) and 87.57% (148/169), respectively (P
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Detection of anti-Plerocercoid IgG antibodies in serum samples of patients with sparganosis and other parasitosis by rSeCP-ELISA and ES-ELISA.
2015Co-Authors: Li Na Liu, Zhong Quan Wang, Xi Zhang, Peng Jiang, Ruo Dan Liu, Zi Fang Zhang, Jing CuiAbstract:Detection of anti-Plerocercoid IgG antibodies in serum samples of patients with sparganosis and other parasitosis by rSeCP-ELISA and ES-ELISA.
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sparganosis henan province central china
Emerging Infectious Diseases, 2011Co-Authors: Jing Cui, Xi Meng Lin, Hong Wei Zhang, Zhong Quan WangAbstract:To the Editor: Sparganosis is a parasitic zoonosis caused by invasion of the spargana, the Plerocercoid larvae of various diphyllobothroid tapeworms belonging to the genus Spirometra (1). Although human sparganosis is cosmopolitan, it is most frequently found in eastern and southeastern Asia (2). During 1927–2009 in the People’s Republic of China, >1,000 cases in humans in 27 provinces were reported; most cases were in southern China, where human infections were mainly acquired by eating raw or insufficiently cooked meat of frogs and snakes or by placing frog or snake flesh on open wounds for treatment of skin ulcers or on eyes to treat inflammation (3,4). Sparganosis is rarely seen in central and northern China. Before 2006, only 3 imported cases from southern China had been reported in Henan Province in central China (5). However, since 2006 in Henan Province, 20 autochthonous cases caused by ingestion of live tadpoles have emerged. To assess the risk for human infection with sparganosis in this province and to strengthen public safety awareness, we investigated spargana infection in the animal hosts of Spirometra tapeworms. During July 2007–July 2010, wild frogs and frog tadpoles were collected from the cities of Shangqiu, Zhoukou, and Luohe in Henan Province. Necropsies identified Plerocercoids in 11.93% (163/1,366) of tadpoles and in 26.58% (172/647) of frogs. By frog species, Plerocercoids were found in 31.09% (111/357) of Rana limmocharis and 26.29% (61/232) of R. nigromaculata frogs, each significantly (p<0.05) more numerous in these species than in R. temporaria frogs (0/58). In addition, 177 wild frogs sold at markets in Luohe were also examined; Plerocercoids were detected in 30.39% (31/102) of R. limmocharis and 28% (21/75) of R. nigromaculata frogs. Thus, in Henan Province, R. limmocharis and R. nigromaculata frogs are the main intermediate hosts of Spirometra tapeworms. No pathologic changes associated with the tapeworms were found in dissected tadpoles and frogs. We found 250 Plerocercoids in tadpoles and 1,387 in frogs. Tadpoles contained 1–14 (mean 1.53) and frogs 1–87 (mean 6.85) tapeworms. In frogs, most Plerocercoids were located in the muscles of hind legs and back; some were in the muscles of the abdominal wall and forelegs. Plerocercoids dissected from tadpole and frog tissues were wrinkled, whitish, and ribbon-shaped and continuously moved while in normal saline. Plerocercoids from tadpoles were 1–8 mm long and 0.2–0.5 mm wide; those from frogs were 1–13 cm long and 1–2.5 mm wide. Cyclops were collected from ponds and ditches by using a 425-µm mesh (no. 40) sieve and species were identified by microscopic appearance as Mesocyclops leuckarti (6,7). Procercoids were microscopically found in the hemocele of 3.53% (3/85) of cyclops; 3–5 worms per cyclop were found. Fecal examination of dogs and cats found Spirometra mansoni tapeworm eggs in 19.35% (6/31) of dogs and 33.33% (1/3) cats. In addition, a 3-month-old specific pathogen–free cat was orally inoculated with 33 Plerocercoids from tadpoles, fecal samples were microscopically examined by sedimentation during 10–25 days postinfection (dpi), and the cat was euthanized and examined for adult worms. S. mansoni tapeworm eggs were found in the feces during 12–25 dpi, and 17 adult worms, 26–45 cm long, were recovered from the small intestines at 25 dpi; however, no Plerocercoids were seen in the tissues. The adult worms were morphologically identified as S. mansoni according to the following features: scolex with 2 longitudinal grooves, mature and gravid proglottids with conspicuous uterus at the center of segments, spiral-shaped uterus (unlike the rosette-shaped uterus of Diphyllobothrium latum), and cirrus and vaginal pore with separate openings (8,9). Although in recent years in Henan Province wild frogs have been sold clandestinely at markets, persons in this province do not routinely eat raw frog meat or use raw meat as poultices. However, some villagers in Henan do believe that eating live tadpoles has a medicinal role for skin diseases and, thus, they contract sparganosis. Accordingly, the route of Plerocercoid infection for humans in Henan Province differs from that in southern China. Because this disease is rare in central and northern China, sparganosis is often neglected and misdiagnosed. Our survey showed that in Henan Province, 11.93% of tadpoles are infected with Plerocercoids and 3.53% of cyclops are infected with procercoids. Therefore, eating live tadpoles poses a high risk for Plerocercoid infection and must be discouraged. In addition, drinking raw water containing cyclops also poses a slight risk for sparganosis.
Motomi Torii - One of the best experts on this subject based on the ideXlab platform.
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the fatty acids of each lipid fraction and their use in providing energy source of the Plerocercoid of spirometra erinacei
International Journal for Parasitology, 1995Co-Authors: Tetsuhito Fukushima, Yosuke Yamane, Akio Isobe, Nobumasa Hojo, Kuninori Shiwaku, Motomi ToriiAbstract:Abstract The fatty acid concentration of each lipid fraction of Plerocercoids of Spirometra erinacei and the host snake serum was investigated. The major fatty acids of phospholipid of the Plerocercoids were C18:1, C18:0 and C16:0, and those of the host snake serum were C16:0, C18:1 and C18:0, in order of amount in both cases. The changes of the fatty acid composition of phospholipid of the Plerocercoids when theywere incubated in physiological saline at 18°C and at 37°C for 24 h were investigated in both cases. Polyunsaturated fatty acids increased at 18°C, and saturated fatty acids increased at 37°C. Michaelis constants ( K m) of beta-hydroxyacyl-CoA dehydrogenase (HAD), NADH: ubiquinone oxidoreductase (complex I) (NADH: ferricyanide reaction) and complex I (NADH: ubiquinone reaction) for NADH were 20.6, 50 and 13.3 μM, respectively. The ATP production in mitochondria of the Plerocercoids was accelerated by adding ADP and inhibited by adding such electron transport system inhibitors as rotenone, antimycin A and sodium cyanide. These results suggested that the fatty acids in the Plerocercoids played an important role in regulating the fluidity of membrane by changing the composition in membrane lipid corresponding with the change of temperature circumstance. The NADH reduced by HAD might be accepted by the complex I in the electron transport system, and thus the parasites were capable of ATP production in a classical pathway of the oxidative phosphorylation system.
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the metabolism of arachidonic acid to prostaglandin e2 in Plerocercoids of spirometra erinacei
Parasitology Research, 1993Co-Authors: Tetsuhito Fukushima, Yosuke Yamane, Akio Isobe, Nobumasa Hojo, K Shiwaku, Motomi ToriiAbstract:With a simplified method of extracting and purifying prostaglandins, trace prostaglandins (nanogram order) were detected by gas chromatography-mass spectrometry. Arachidonic acid was metabolized to prostaglandin E2 (PGE2) by Plerocercoids ofSpirometra erinacei, the PGE2 was detected in the medium after incubation with arachidonic acid, and the role of albumin in the absorption of free arachidonic acid by Plerocercoids and in the release of its metabolite was investigated. Plerocercoids absorbed arachidonic acid-binding albumin and released PGE2 efficiently. PGE2 is known to suppress the functions of mononuclear cells of the host. The selective release of PGE2 may be related to the escape mechanism of Plerocercoids ofS. erinacei from the host immune system to become established larva migrans, i.e., sparganosis.
A S Clarke - One of the best experts on this subject based on the ideXlab platform.
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studies on the life cycle of the pseudophyllidean cestode schistocephalus solidus
Journal of Zoology, 2010Co-Authors: A S ClarkeAbstract:Summary 1A study of host-parasite relationships has been made on the cestode Schistocephalus solidus Muller by examining conditions relating to the infection and growth of the Plerocercoid in the stickleback, and- its transfer to other hosts for maturation; the infection of Cyclops by the coracidimn with its subsequent change to a procercoid; and the passage of the procercoid into the body cavity of the fish to become a Plerocercoid. It has been possible, by establishing suitable techniques, to carry out the whole life cycle in the laboratory and so facilitate these studies. 2Eggs were obtained from Plerocercoids and observations made on their development. The time of hatching of the coracidium varies considerably, both with temperature and with intrinsic factors. The average time is about three weeks but hatching may occur up to six months after the eggs have been shed. 3From such eggs, free coracidia and, subsequently, procercoids in Cyclops and nauplii, were secured and studied. The growth rate of procercoids was determined together with the effect of infection on the growth of nauplii. 4Infections in fish were secured from infected Cyclops but not in sufficient numbers to justify more than tentative conclusions about the growth rate of Plerocercoids. The time required for penetration of the fish gut wall by the procercoid can be as short as two hours. 5Changes in morphology of the worm in its transformation from a coracidium to a Plerocercoid were surveyed and it was shown how the formation of the musculature precedes the initiation of strobilation which occurs rapidly and completely, followed immediately by the formation of the genital rudiments. 6The excretory system of the Plerocercoid could be displayed effectively by a method of embedding in transparent plastic. 7Infected fish from one source were collected throughout the year and the weights of the fish and of the worms within them were ascertained. It was found that in the autumn there was an increase in the number of large fish in the marginal zone–possibly related to an increase of plankton–but during the months from winter to summer the mean size of fish was less and remained relatively stable in this zone. The mean weights of Plerocercoids however, increased during this period as did the temperature of the water. Some indications of the growth rate of the Plerocercoid were observed from the collected data. 8An examination of the stomach contents of the fish showed a wide variety of diet with little or no change throughout the year and also indicated that Cyclops form an almost insignificant feature in this diet.
Kirk Phares - One of the best experts on this subject based on the ideXlab platform.
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an unusual host parasite relationship the growth hormone like factor from Plerocercoids of spirometrid tapeworms
International Journal for Parasitology, 1996Co-Authors: Kirk PharesAbstract:Abstract Accelerated body growth is associated with experimental infections with the Plerocercoid stage of the pseudophyllidean tapeworm, Spirometra mansonoides . The growth response is due to a protein that is synthesized and released by Plerocercoids in the host. Plerocercoid growth factor (PGF) is transported by the blood, interacts with growth hormone (GH) receptors and mimics many of the biological actions of GH. Purification and characterization of PGF reveals that the GH-like factor is a 27.5 kDa cysteine proteinase. Sequencing of a cDNA for PGF revealed 40–50% homology to mammalian cysteine proteinases, but no homology to any GH. The GH-like factor/proteinase is found in abundance in the tegument of Plerocercoids where the proteinase activity would be most effective in aiding tissue invasion, and possibly evasion, of the host's immune response to the parasite. The advantage of the GH-like activity is less clear, but it may affect the species survival by suppressing specific elements of the host's immune system and by improving the probability that the infected host will be eaten by a definitive host by increasing morbidity and/or mortality, conditions which are associated with long-term exposure to elevated levels of GH and PGF.
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the growth factor from Plerocercoids of spirometra mansonoides is both a growth hormone agonist and a cysteine proteinase
Journal of Parasitology, 1996Co-Authors: Kirk Phares, Jacy KubikAbstract:Plerocercoids of the tapeworm Spirometra mansonoides produce a substance that stimulates growth of experimental hosts. We report purification of Plerocercoid growth factor (PGF) to homogeneity by a process involving isolation and solubilization of Plerocercoid membranes, isoelectric point selection by chromatofocusing chromatography or preparative isoelectric focusing, and anion-exchange chromatography. A radioreceptor assay (RRA) for human growth hormone (hGH) was used to detect PGF and purity of the 27.5-kDa protein was judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Proteolytic activity was detected in the 27.5-kDa protein by gelatin substrate PAGE. Characterization of PGF as a neutral cysteine proteinase was based on substrate and inhibitor specificities and dependence on pH and thiol-containing reagents. The association of hGH agonist and proteinase activities was shown by comparing RRA and hydrolytic activities in the presence and absence of the cysteine proteinase inhibitor E-64.