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Sample records for block schistosoma japonicum

  1. Taxonomy Icon Data: Schistosoma japonicum [Taxonomy Icon

    Lifescience Database Archive (English)

    Full Text Available .png Schistosoma_japonicum_NL.png Schistosoma_japonicum_S.png Schistosoma_japonicum_NS.png http://biosciencedbc.jp/taxonomy..._icon/icon.cgi?i=Schistosoma+japonicum&t=L http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=S...chistosoma+japonicum&t=NL http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Schist...osoma+japonicum&t=S http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Schistosoma+japonicum&t=NS http://togodb.biosciencedbc.jp/togodb/view/taxonomy_icon_comment_en?species_id=132 ...

  2. Non-equilibrium plasma prevention of Schistosoma japonicum transmission.

    Science.gov (United States)

    Wang, Xing-Quan; Wang, Feng-Peng; Chen, Wei; Huang, Jun; Bazaka, Kateryna; Ostrikov, Kostya Ken

    2016-10-14

    Schistosoma japonicum is a widespread human and animal parasite that causes intestinal and hepatosplenic schistosomiasis linked to colon, liver and bladder cancers, and anemia. Estimated 230 million people are currently infected with Schistosoma spp, with 779 million people at risk of contracting the parasite. Infection occurs when a host comes into contact with cercariae, a planktonic larval stage of the parasite, and can be prevented by inactivating the larvae, commonly by chemical treatment. We investigated the use of physical non-equilibrium plasma generated at atmospheric pressure using custom-made dielectric barrier discharge reactor to kill S. japonicum cercariae. Survival rate decreased with treatment time and applied power. Plasmas generated in O2 and air gas discharges were more effective in killing S. japonicum cercariae than that generated in He, which is directly related to the mechanism by which cercariae are inactivated. Reactive oxygen species, such as O atoms, abundant in O2 plasma and NO in air plasma play a major role in killing of S. japonicum cercariae via oxidation mechanisms. Similar level of efficacy is also shown for a gliding arc discharge plasma jet generated in ambient air, a system that may be more appropriate for scale-up and integration into existing water treatment processes.

  3. Non-equilibrium plasma prevention of Schistosoma japonicum transmission

    Science.gov (United States)

    Wang, Xing-Quan; Wang, Feng-Peng; Chen, Wei; Huang, Jun; Bazaka, Kateryna; Ostrikov, Kostya (Ken)

    2016-10-01

    Schistosoma japonicum is a widespread human and animal parasite that causes intestinal and hepatosplenic schistosomiasis linked to colon, liver and bladder cancers, and anemia. Estimated 230 million people are currently infected with Schistosoma spp, with 779 million people at risk of contracting the parasite. Infection occurs when a host comes into contact with cercariae, a planktonic larval stage of the parasite, and can be prevented by inactivating the larvae, commonly by chemical treatment. We investigated the use of physical non-equilibrium plasma generated at atmospheric pressure using custom-made dielectric barrier discharge reactor to kill S. japonicum cercariae. Survival rate decreased with treatment time and applied power. Plasmas generated in O2 and air gas discharges were more effective in killing S. japonicum cercariae than that generated in He, which is directly related to the mechanism by which cercariae are inactivated. Reactive oxygen species, such as O atoms, abundant in O2 plasma and NO in air plasma play a major role in killing of S. japonicum cercariae via oxidation mechanisms. Similar level of efficacy is also shown for a gliding arc discharge plasma jet generated in ambient air, a system that may be more appropriate for scale-up and integration into existing water treatment processes.

  4. Transmission dynamics of Schistosoma japonicum in the lakes and marshlands of China.

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    Darren J Gray

    Full Text Available BACKGROUND: Schistosoma japonicum is a major public health concern in China, with over one million people infected and another 40 million living in areas at risk of infection. Unlike the disease caused by S. mansoni and S. haematobium, schistosomiasis japonica is a zoonosis, involving a number of different mammalian species as reservoir hosts. As a result of a number of published reports from China, it has long been considered that bovines, particularly water buffaloes, play a major role in human S. japonicum transmission there, and a drug-based intervention study (1998-2003 around the Poyang Lake in Jiangxi Province provided proof of concept that water buffaloes are, indeed, major reservoirs of human infection in this setting. METHODS AND FINDINGS: In this study we incorporated recently obtained epidemiological information to model the steady-state S. japonicum transmission as well as the impact of the removal of S. japonicum transmission attributable to water buffaloes on human infection rates across six different endemic scenarios within three villages in the Dongting (Hunan and Poyang (Jiangxi lakes of southern China. Similar results were obtained for all scenarios. Steady-state S. japonicum infection rates remained constant and human prevalence and incidence were predicted to fall considerably over time. The model showed that the contribution of S. japonicum water buffalo transmission to human infection ranged from 39.1% to 99.1% and predicted that the removal of water buffalo transmission would reduce parasite reproductive rates below 1. This indicates that without the contribution of water buffaloes, S. japonicum transmission is interrupted and unsustainable. These scenarios are generalizable to other endemic villages in the lake and marshland areas of China where a similar cycle of snail infection and infection/reinfection of humans and bovines occurs. CONCLUSIONS: Along with previous epidemiological data, our findings strongly support

  5. A REVIEW OF SCHISTOSOMIASIS IN INDONESIA WITH REFERENCE TO SCHISTOSOMA JAPONICUM

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    Lim Boo Liat

    2012-09-01

    Full Text Available Tinjauan tentang Schistosoma di Indonesia ini mencakup Schistosoma japonicum, S. incognitum, S. spindale dan Trichohilharzia brevis. Tinjauan dibuat atas dasar laporan-laporan penelitian yang telah diterbitkan. Di dalamnya dapat dijumpai uraian singkat tentang S. spinale dan T. brevis. Dari banyak publikasi tentang S. japonicum dan S. incoganitum dapat disajikan uraian tentang peranan kedua parasit tersebut sebagai penyebab penyakit baik manusia maupun hewan.

  6. Schistosoma japonicum-associated morbidity and its mechanisms among children and young adults in the Philippines

    NARCIS (Netherlands)

    Coutinho, H.M.

    2006-01-01

    This thesis presents results from a longitudinal study in Schistosoma japonicum infected children, adolescents and young adults, carried out in the Philippines. The aim of this thesis was to describe nutritional morbidity and hepatic fibrosis in the context of S. japonicum infection and reinfection

  7. Cloning and characterisation of Schistosoma japonicum insulin receptors.

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    Hong You

    Full Text Available BACKGROUND: Schistosomes depend for growth and development on host hormonal signals, which may include the insulin signalling pathway. We cloned and assessed the function of two insulin receptors from Schistosoma japonicum in order to shed light on their role in schistosome biology. METHODOLOGY/PRINCIPAL FINDINGS: We isolated, from S. japonicum, insulin receptors 1 (SjIR-1 and 2 (SjIR-2 sharing close sequence identity to their S. mansoni homologues (SmIR-1 and SmIR-2. SjIR-1 is located on the tegument basal membrane and the internal epithelium of adult worms, whereas SjIR-2 is located in the parenchyma of males and the vitelline tissue of females. Phylogenetic analysis showed that SjIR-2 and SmIR-2 are close to Echinococcus multilocularis insulin receptor (EmIR, suggesting that SjIR-2, SmIR-2 and EmIR share similar roles in growth and development in the three taxa. Structure homology modelling recovered the conserved structure between the SjIRs and Homo sapiens IR (HIR implying a common predicted binding mechanism in the ligand domain and the same downstream signal transduction processing in the tyrosine kinase domain as in HIR. Two-hybrid analysis was used to confirm that the ligand domains of SjIR-1 and SjIR-2 contain the insulin binding site. Incubation of adult worms in vitro, both with a specific insulin receptor inhibitor and anti-SjIRs antibodies, resulted in a significant decrease in worm glucose levels, suggesting again the same function for SjIRs in regulating glucose uptake as described for mammalian cells. CONCLUSIONS: Adult worms of S. japonicum possess insulin receptors that can specifically bind to insulin, indicating that the parasite can utilize host insulin for development and growth by sharing the same pathway as mammalian cells in regulating glucose uptake. A complete understanding of the role of SjIRs in the biology of S. japonicum may result in their use as new targets for drug and vaccine development against

  8. Biology and Control of Snail Intermediate Host of Schistosoma japonicum in The People's Republic of China

    DEFF Research Database (Denmark)

    Li, Z.J.; Ge, J; Dai, J.R.

    2016-01-01

    Schistosomiasis caused by Schistosoma japonicum is a severe parasitic disease in The People's Republic of China and imposed considerable burden on human and domestic animal health and socioeconomic development. The significant achievement in schistosomiasis control has been made in last 60years. ....... Oncomelania hupensis as the only intermediate host of S. japonicum plays a key role in disease transmission. The habitat complexity of the snails challenges to effective control. In this review we share the experiences in control and research of O. hupensis....

  9. Developmental gene expression profiles of the human pathogen Schistosoma japonicum

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    McManus Donald P

    2009-03-01

    Full Text Available Abstract Background The schistosome blood flukes are complex trematodes and cause a chronic parasitic disease of significant public health importance worldwide, schistosomiasis. Their life cycle is characterised by distinct parasitic and free-living phases involving mammalian and snail hosts and freshwater. Microarray analysis was used to profile developmental gene expression in the Asian species, Schistosoma japonicum. Total RNAs were isolated from the three distinct environmental phases of the lifecycle – aquatic/snail (eggs, miracidia, sporocysts, cercariae, juvenile (lung schistosomula and paired but pre-egg laying adults and adult (paired, mature males and egg-producing females, both examined separately. Advanced analyses including ANOVA, principal component analysis, and hierarchal clustering provided a global synopsis of gene expression relationships among the different developmental stages of the schistosome parasite. Results Gene expression profiles were linked to the major environmental settings through which the developmental stages of the fluke have to adapt during the course of its life cycle. Gene ontologies of the differentially expressed genes revealed a wide range of functions and processes. In addition, stage-specific, differentially expressed genes were identified that were involved in numerous biological pathways and functions including calcium signalling, sphingolipid metabolism and parasite defence. Conclusion The findings provide a comprehensive database of gene expression in an important human pathogen, including transcriptional changes in genes involved in evasion of the host immune response, nutrient acquisition, energy production, calcium signalling, sphingolipid metabolism, egg production and tegumental function during development. This resource should help facilitate the identification and prioritization of new anti-schistosome drug and vaccine targets for the control of schistosomiasis.

  10. Congenital infection of rabbits with Schistosoma japonicum and protective immunity of offspring

    Institute of Scientific and Technical Information of China (English)

    SHI You-en 石佑恩; Abdel-Moneim M Salim 沙里模; NING Chang-xiu 宁长修; GAN Yan 甘燕; ZHU Xiao-hua 朱晓华; PU Ling-yi 卜玲毅

    2004-01-01

    Background Recently congenital infection with Schistosoma japonicum (S. Japonicum) has been domonstrated in pigs, rabbits, mice and dogs. We explored the rabbit as an animal model for the congenital infection of schistosomiasis japonica and assessed the effect of a congenital S. Japonicum infection on the resistance of rabbit kittens to a postnatal challenge infection.Methods Sixteen pregnant New Zealand white rabbits were infected with a single dose of S. Japonicum cercariae. The exposed animals were divided into three groups according to the gestation age at the time of infection. Diagnosis of prenatally acquired S. Japonicum infection in the rabbit kittens was primarily based on serological tests in combination with parasitological and histopathological findings. Congenitally infected kittens were challenged percutaneously with 100 S. Japonicum cercariae to assess the effect of a congenital S. Japonicum infection on kitten resistance to a postnatal challenge infection.Conclusions This study demonstrates the possibility of congenital infection of S. Japonicum in rabbits and the resistance of congenitally infected kittens to a postnatal challenge infection. These results have important implications not only for epidemiological investigations, but also in designing government control programs for schistosomiasis.

  11. Identification of Host Insulin Binding Sites on Schistosoma japonicum Insulin Receptors.

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    Rachel J Stephenson

    Full Text Available Schistosoma japonicum insulin receptors (SjIRs have been identified as encouraging vaccine candidates. Interrupting or blocking the binding between host insulin and the schistosome insulin receptors (IRs may result in reduced glucose uptake leading to starvation and stunting of worms with a reduction in egg output. To further understand how schistosomes are able to exploit host insulin for development and growth, and whether these parasites and their mammalian hosts compete for the same insulin source, we identified insulin binding sites on the SjIRs. Based on sequence analysis and the predicted antigenic structure of the primary sequences of the SjIRs, we designed nine and eleven peptide analogues from SjIR-1 and SjIR-2, respectively. Using the Octet RED system, we identified analogues derived from SjIR-1 (10 and SjIR-2 (20, 21 and 22 with insulin-binding sequences specific for S. japonicum. Nevertheless, the human insulin receptor (HIR may compete with the SjIRs in binding human insulin in other positions which are important for HIR binding to insulin. However, no binding occurred between insulin and parasite analogues derived from SjIR-1 (2, 7 and 8 and SjIR-2 (14, 16 and 18 at the same locations as HIR sequences which have been shown to have strong insulin binding affinities. Importantly, we found two analogues (1 and 3, derived from SjIR-1, and two analogues (13 and 15 derived from SjIR-2, were responsible for the major insulin binding affinity in S. japonicum. These peptide analogues were shown to have more than 10 times (in KD value stronger binding capacity for human insulin compared with peptides derived from the HIR in the same sequence positions. Paradoxically, analogues 1, 3, 13 and 15 do not appear to contain major antigenic determinants which resulted in poor antibody responses to native S. japonicum protein. This argues against their future development as peptide-vaccine candidates.

  12. Constructing the recombinant plasmid-pLXSN-SjYF-of Yolk Ferritin gene of Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    ZhouJunmei; YuXinbing; WuZhongdao; ZhengYinan; LiYan

    1999-01-01

    TO construct the recotabinant plasmid —— pLXSN-SjYF to prepare for expression and DNA vaccine of Schistosoma japonicum gene. Methods :Mnpliffing DNA fragment coding yolk fereltin from a female adult Sehistosoma japonicum DNAs by PCR. The fragment was inseted into pLXSN retrovirus vector by digesting with restrictive enzymes and linking reactions. The positive clone was screened on LB plates contmaining amplcillin asld identified by restrictive enzymes digestion and PCR amplification, Resttlts The specific DNA fragmem SjYF was amplified from the female adult SjDNAs. IaLXSN-SjYF was constructed successfully and the further research will be carried out.

  13. Does multiple hosts mean multiple parasites? Population genetic structure of Schistosoma japonicum between definitive host species.

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    Wang, T P; Shrivastava, J; Johansen, M V; Zhang, S Q; Wang, F F; Webster, J P

    2006-10-01

    Multi-host parasites, those capable of infecting more than one species of host, are responsible for the majority of all zoonotic, emerging or persistent human and animal diseases and are considered one of the major challenges for the biomedical sciences in the 21st century. We characterized the population structure of the multi-host parasite Schistosoma japonicum in relation to its definitive host species by genotyping miracidia collected from humans and domestic animals across five villages around the Yangtze River in Anhui Province, mainland China, using microsatellite markers. High levels of polymorphisms were observed and two main genetic clusters were identified which separated water buffalo, cattle and humans from goats, pigs, dogs and cats. We thereby believe that we present the first evidence of definitive host-based genetic variation in Schistosoma japonicum which has important epidemiological, evolutionary, medical and veterinary implications.

  14. Suppression of the Insulin Receptors in Adult Schistosoma japonicum Impacts on Parasite Growth and Development: Further Evidence of Vaccine Potential.

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    You, Hong; Gobert, Geoffrey N; Cai, Pengfei; Mou, Rong; Nawaratna, Sujeevi; Fang, Guofu; Villinger, Francois; McManus, Donald P

    2015-05-01

    To further investigate the importance of insulin signaling in the growth, development, sexual maturation and egg production of adult schistosomes, we have focused attention on the insulin receptors (SjIRs) of Schistosoma japonicum, which we have previously cloned and partially characterised. We now show, by Biolayer Interferometry, that human insulin can bind the L1 subdomain (insulin binding domain) of recombinant (r)SjIR1 and rSjIR2 (designated SjLD1 and SjLD2) produced using the Drosophila S2 protein expression system. We have then used RNA interference (RNAi) to knock down the expression of the SjIRs in adult S. japonicum in vitro and show that, in addition to their reduced transcription, the transcript levels of other important downstream genes within the insulin pathway, associated with glucose metabolism and schistosome fecundity, were also impacted substantially. Further, a significant decrease in glucose uptake was observed in the SjIR-knockdown worms compared with luciferase controls. In vaccine/challenge experiments, we found that rSjLD1 and rSjLD2 depressed female growth, intestinal granuloma density and faecal egg production in S. japonicum in mice presented with a low dose challenge infection. These data re-emphasize the potential of the SjIRs as veterinary transmission blocking vaccine candidates against zoonotic schistosomiasis japonica in China and the Philippines.

  15. Identification and Characterization of Peptides Mimicking the Epitopes of Metalloprotease of Schistosoma Japonicum

    Institute of Scientific and Technical Information of China (English)

    Lianfei Tang; Yuxiao Chen; Linqian Wang; Shunke Zhang; Xianfang Zeng; Xinyuan Yi

    2005-01-01

    In an attempt to isolate and characterize peptides mimicking epitopes of metalloprotease and explore their immunological protection against Schistosoma japonicum (S. japonicum), polyclonal anti-metalloprotease sera was prepared to screen a 12-mer random peptide library to isolate phages binding specially to antisera IgG. Then,phage ELISA, animal immunization, DNA sequencing, Western blotting and enzymatic activity neutralizing analysis were used to characterize the selected phage clones. All of ten randomly picked clones were shown to be positive. Five peptides of different amino acid sequences deduced from DNA sequences were obtained and two of them (peptides 2 and 3) could induce significant reduction (31.0% and 31.8%, respectively) in worm burden and high reduction (52.6% and 54.9%, respectively) in liver eggs per gram (LEPG), while, unexpectedly, others (peptides 1, 4 and 5) could not elicit enough protection against infection of S. japonicum. Peptides 2 and 3 could be recognized by S. japonicum infected mouse sera (IMS) and could elicit neutralizing Abs. The results show that peptides 2 and 3 are antigenic and immunogenic. They are true mimics of epitopes of metalloprotease and useful as novel vaccine candidates against S. japonicum.

  16. Partial protection induced by phage library-selected peptides mimicking epitopes of Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    Objective To obtain peptide mimicking epitopes of Schistosoma japonicum (S.japonicum) through screening of a phage peptide library and to test their potential for induction of protection. Methods S.japonicum infected sera from Microtus fortis (IMFS) and normal sera from Microtus fortis (NMFS) were used respectively to screen a 12-mers random peptide library by testing the reactivity of anti-S.japonicum serum with the phagotopes. After three rounds of biopanning, the pooled phages were used to immunize mice, after which challenge infection was performed. Results Of 12 randomly picked clones, 10 clones selected using IMFS and 7 clones selected using NMFS were shown to be antigenic. Significant reduction in adult worms (22.6%) and a high reduction (68.9%) in liver eggs were achieved following immunization with phages screened with IMFS. However, no protection was elicited by those selected with NMFS. Conclusion The results show that the phagotopes are both antigenic and immunogenic, suggesting a potential use of phage displayed peptide as novel vaccines against S. japonicum.

  17. New perspectives on host-parasite interplay by comparative transcriptomic and proteomic analyses of Schistosoma japonicum.

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    Feng Liu

    2006-04-01

    Full Text Available Schistosomiasis remains a serious public health problem with an estimated 200 million people infected in 76 countries. Here we isolated ~ 8,400 potential protein-encoding cDNA contigs from Schistosoma japonicum after sequencing circa 84,000 expressed sequence tags. In tandem, we undertook a high-throughput proteomics approach to characterize the protein expression profiles of a number of developmental stages (cercariae, hepatic schistosomula, female and male adults, eggs, and miracidia and tissues at the host-parasite interface (eggshell and tegument by interrogating the protein database deduced from the contigs. Comparative analysis of these transcriptomic and proteomic data, the latter including 3,260 proteins with putative identities, revealed differential expression of genes among the various developmental stages and sexes of S. japonicum and localization of putative secretory and membrane antigens, enzymes, and other gene products on the adult tegument and eggshell, many of which displayed genetic polymorphisms. Numerous S. japonicum genes exhibited high levels of identity with those of their mammalian hosts, whereas many others appeared to be conserved only across the genus Schistosoma or Phylum Platyhelminthes. These findings are expected to provide new insights into the pathophysiology of schistosomiasis and for the development of improved interventions for disease control and will facilitate a more fundamental understanding of schistosome biology, evolution, and the host-parasite interplay.

  18. New Perspectives on Host-Parasite Interplay by Comparative Transcriptomic and Proteomic Analyses of Schistosoma japonicum.

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    2006-04-01

    Full Text Available Schistosomiasis remains a serious public health problem with an estimated 200 million people infected in 76 countries. Here we isolated ~ 8,400 potential protein-encoding cDNA contigs from Schistosoma japonicum after sequencing circa 84,000 expressed sequence tags. In tandem, we undertook a high-throughput proteomics approach to characterize the protein expression profiles of a number of developmental stages (cercariae, hepatic schistosomula, female and male adults, eggs, and miracidia and tissues at the host-parasite interface (eggshell and tegument by interrogating the protein database deduced from the contigs. Comparative analysis of these transcriptomic and proteomic data, the latter including 3,260 proteins with putative identities, revealed differential expression of genes among the various developmental stages and sexes of S. japonicum and localization of putative secretory and membrane antigens, enzymes, and other gene products on the adult tegument and eggshell, many of which displayed genetic polymorphisms. Numerous S. japonicum genes exhibited high levels of identity with those of their mammalian hosts, whereas many others appeared to be conserved only across the genus Schistosoma or Phylum Platyhelminthes. These findings are expected to provide new insights into the pathophysiology of schistosomiasis and for the development of improved interventions for disease control and will facilitate a more fundamental understanding of schistosome biology, evolution, and the host-parasite interplay.

  19. Homology-based annotation of non-coding RNAs in the genomes of Schistosoma mansoni and Schistosoma japonicum

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    Santana Clara

    2009-10-01

    Full Text Available Abstract Background Schistosomes are trematode parasites of the phylum Platyhelminthes. They are considered the most important of the human helminth parasites in terms of morbidity and mortality. Draft genome sequences are now available for Schistosoma mansoni and Schistosoma japonicum. Non-coding RNA (ncRNA plays a crucial role in gene expression regulation, cellular function and defense, homeostasis, and pathogenesis. The genome-wide annotation of ncRNAs is a non-trivial task unless well-annotated genomes of closely related species are already available. Results A homology search for structured ncRNA in the genome of S. mansoni resulted in 23 types of ncRNAs with conserved primary and secondary structure. Among these, we identified rRNA, snRNA, SL RNA, SRP, tRNAs and RNase P, and also possibly MRP and 7SK RNAs. In addition, we confirmed five miRNAs that have recently been reported in S. japonicum and found two additional homologs of known miRNAs. The tRNA complement of S. mansoni is comparable to that of the free-living planarian Schmidtea mediterranea, although for some amino acids differences of more than a factor of two are observed: Leu, Ser, and His are overrepresented, while Cys, Meth, and Ile are underrepresented in S. mansoni. On the other hand, the number of tRNAs in the genome of S. japonicum is reduced by more than a factor of four. Both schistosomes have a complete set of minor spliceosomal snRNAs. Several ncRNAs that are expected to exist in the S. mansoni genome were not found, among them the telomerase RNA, vault RNAs, and Y RNAs. Conclusion The ncRNA sequences and structures presented here represent the most complete dataset of ncRNA from any lophotrochozoan reported so far. This data set provides an important reference for further analysis of the genomes of schistosomes and indeed eukaryotic genomes at large.

  20. Genome-wide identification of Schistosoma japonicum microRNAs using a deep-sequencing approach.

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    Jian Huang

    Full Text Available BACKGROUND: Human schistosomiasis is one of the most prevalent and serious parasitic diseases worldwide. Schistosoma japonicum is one of important pathogens of this disease. MicroRNAs (miRNAs are a large group of non-coding RNAs that play important roles in regulating gene expression and protein translation in animals. Genome-wide identification of miRNAs in a given organism is a critical step to facilitating our understanding of genome organization, genome biology, evolution, and posttranscriptional regulation. METHODOLOGY/PRINCIPAL FINDINGS: We sequenced two small RNA libraries prepared from different stages of the life cycle of S. japonicum, immature schistosomula and mature pairing adults, through a deep DNA sequencing approach, which yielded approximately 12 million high-quality short sequence reads containing a total of approximately 2 million non-redundant tags. Based on a bioinformatics pipeline, we identified 176 new S. japonicum miRNAs, of which some exhibited a differential pattern of expression between the two stages. Although 21 S. japonicum miRNAs are orthologs of known miRNAs within the metazoans, some nucleotides at many positions of Schistosoma miRNAs, such as miR-8, let-7, miR-10, miR-31, miR-92, miR-124, and miR-125, are indeed significantly distinct from other bilaterian orthologs. In addition, both miR-71 and some miR-2 family members in tandem are found to be clustered in a reversal direction model on two genomic loci, and two pairs of novel S. japonicum miRNAs were derived from sense and antisense DNA strands at the same genomic loci. CONCLUSIONS/SIGNIFICANCE: The collection of S. japonicum miRNAs could be used as a new platform to study the genomic structure, gene regulation and networks, evolutionary processes, development, and host-parasite interactions. Some S. japonicum miRNAs and their clusters could represent the ancestral forms of the conserved orthologues and a model for the genesis of novel miRNAs.

  1. Follicular helper T cells promote liver pathology in mice during Schistosoma japonicum infection.

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    Xiaojun Chen

    2014-05-01

    Full Text Available Following Schistosoma japonicum (S. japonicum infection, granulomatous responses are induced by parasite eggs trapped in host organs, particular in the liver, during the acute stage of disease. While excessive liver granulomatous responses can lead to more severe fibrosis and circulatory impairment in chronically infected host. However, the exact mechanism of hepatic granuloma formation has remained obscure. In this study, we for the first time showed that follicular helper T (Tfh cells are recruited to the liver to upregulate hepatic granuloma formation and liver injury in S. japonicum-infected mice, and identified a novel function of macrophages in Tfh cell induction. In addition, our results showed that the generation of Tfh cells driven by macrophages is dependent on cell-cell contact and the level of inducible costimulator ligand (ICOSL on macrophages which is regulated by CD40-CD40L signaling. Our findings uncovered a previously unappreciated role for Tfh cells in liver pathology caused by S. japonicum infection in mice.

  2. Co-dispersal of the blood fluke Schistosoma japonicum and Homo sapiens in the Neolithic Age.

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    Yin, Mingbo; Zheng, Hong-Xiang; Su, Jing; Feng, Zheng; McManus, Donald P; Zhou, Xiao-Nong; Jin, Li; Hu, Wei

    2015-12-21

    The global spread of human infectious diseases is of considerable public health and biomedical interest. Little is known about the relationship between the distribution of ancient parasites and that of their human hosts. Schistosoma japonicum is one of the three major species of schistosome blood flukes causing the disease of schistosomiasis in humans. The parasite is prevalent in East and Southeast Asia, including the People's Republic of China, the Philippines and Indonesia. We studied the co-expansion of S. japonicum and its human definitive host. Phylogenetic reconstruction based on complete mitochondrial genome sequences showed that S. japonicum radiated from the middle and lower reaches of the Yangtze River to the mountainous areas of China, Japan and Southeast Asia. In addition, the parasite experienced two population expansions during the Neolithic agriculture era, coinciding with human migration and population growth. The data indicate that the advent of rice planting likely played a key role in the spread of schistosomiasis in Asia. Moreover, the presence of different subspecies of Oncomelania hupensis intermediate host snails in different localities in Asia allowed S. japonicum to survive in new rice-planting areas, and concurrently drove the intraspecies divergence of the parasite.

  3. Schistosoma japonicum: An ultraviolet-attenuated cercarial vaccine applicable in the field for water buffaloes

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    Shi, Y.E.; Jiang, C.F.; Han, J.J.; Li, Y.L.; Ruppel, A. (Tongii Medical Univ., Wuhan, Hubei Province (China))

    1990-07-01

    Water buffaloes were vaccinated three times with 10,000 Schistosoma japonicum cercariae irradiated with ultraviolet (uv) light at a dose of 400 microW x min/cm2. The irradiation was performed with cheap, simple, and portable equipment in a rural area of Hubei Province (People's Republic of China). A challenge infection of 1000 untreated cercariae was given to six vaccinated and six naive control buffaloes, while two vaccinated animals were not challenged. The experiment was terminated 6 weeks after the challenge. Control animals had lost body weight and harbored a mean of 110 worms and 37 eggs per gram of liver. The vaccinated animals gained weight after the challenge and developed 89% resistance to infection with S. japonicum. Since schistosomiasis japonica is nowadays transmitted in China predominantly by domestic livestock, a uv-attenuated cercarial vaccine for bovines may contribute to the control of this disease.

  4. Antischistosomal activity of N,N'-arylurea analogs against Schistosoma japonicum.

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    Yao, Houzong; Liu, Fengyou; Chen, Jinglei; Li, Yan; Cui, Jinhao; Qiao, Chunhua

    2016-03-01

    Although the antischistosomal activities of N,N'-arylurea analogs were reported, systematic structure-activity relationships have not been conducted. In this Letter, we reported the design, synthesis and evaluation of 45 N,N'-arylurea analogs. Among these prepared compounds, 13 compounds were urea linker modified and 32 were N,N'-arylurea derivatives. The activity evaluation revealed 12 analogs exhibited IC50 values lower than 22.6μM, and 7 of them had IC50 less than 10μM against the juvenile Schistosoma japonicum in vitro. Their worm killing potency was even higher against adult worm. Unfortunately, low to moderate worm burden reduction of 0-33.4% was recorded after administration of a single oral dose of 200mg/kg or 400mg/kg to mice harboring S. japonicum.

  5. Cytokine mRNA profiles in pigs exposed prenatally and postnatally to Schistosoma japonicum

    DEFF Research Database (Denmark)

    Techau, Michala E.; Johansen, Maria V.; Aasted, Bent;

    2007-01-01

    The pig is a natural host for Schistosoma japonicum and a useful animal model of human infection. The aim of the present study was to assess the differences between the cytokine profiles in prenatally or postnatally S. japonicum exposed pigs. Seven prenatally exposed pigs, 7 postnatally exposed...... pigs and 4 uninfected control pigs were compared 27 weeks post exposure. Variables included worm burdens, tissue egg counts, liver pathology and mRNA levels of IL-2, IL-4, IL-10, IL-12, TNF-alpha, TGF-beta 1 and IFN-gamma in the liver and the caecum, assessed by RT-PCR. Infection intensity and level...... of septal fibrosis were significantly higher in the postnatal group compared to the prenatal group (P TNF-alpha (P

  6. Induction of species-specific immunity against Schistosoma japonicum by exposure of rats to ultra-violet attenuated cercariae

    Energy Technology Data Exchange (ETDEWEB)

    Moloney, N.A.; Webbe, G.; Hinchcliffe, P.

    1987-02-01

    Single percutaneous immunizations of Fischer rats with 1000 ultra-violet attenuated Schistosoma japonicum cercariae induced 52-88% resistance to challenge 4 weeks later. Increasing this to 3 immunizations induced 90% resistance to challenge, and this level of protection remained undiminished for up to 40 weeks after vaccination. Rats vaccinated with gamma-irradiated S. mansoni cercariae were resistant to challenge with S. mansoni but not S. japonicum. Similarly rats vaccinated with u.v.-attenuated S. japonicum cercariae were not resistant to heterologous challenge. Thus irradiated vaccines are species-specific in both permissive and non-permissive hosts.

  7. Immunization of mice with cells from juvenile worms of Schistosoma japonicum provides immunoprotection against schistosomiasis

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    To validate the protective efficacy against schistosomiasis by immunization with cells from juvenile Schistosoma japonicum in a murine model and to analyze possible factors related to protection, in this study, two independent repeated vaccination trials were performed. After three subcutaneous vaccinations, in trial one, in the absence of adjuvant, primary juvenile worm cells (pJCs) from S. japonicum induced remarkable average reductions in worm burden (54.3%), liver eggs per gram (LEPG) load (59.8%) as well as egg granulomas size (66.5%) compared to PBS control group (P<0.01), which were significantly higher than those elicited by fractions of juvenile worm cells (JCFs) or fractions of juvenile worms (JWFs) (P<0.05). Non-cell components of worms (WNCs) showed no significant protection. In trial two, compared to PBS control group, significant protective effect was also observed for cultured juvenile worm cells (cJCs) from S. japonicum with 58.4% worm reduction and 68.1% LEPG reduction (P<0.01). However, cultured adult worms cells (cACs) showed significantly higher worm burden (P<0.05) and egg burden (P<0.01) when compared to cJCs. Immunological analysis of trial two revealed that cJCs engendered a Th1-biased mixed Th1/Th2 type of immune response while cACs elicited a Th2-type response. Our data indicated that immunization with both primary and cultured cells from S. japonicum juvenile worms provided high immunoprotection, for which the physical character of immunogens, stage-specific parasite and the type of immune response induced might be responsible, suggesting that vaccination with whole cells from S. japonicum larvae is a promising approach to produce protective immunity against schistosomiasis.

  8. A circular analysis of chronobiology of Schistosoma japonicum cercarial emergence from hilly areas of Anhui, China.

    Science.gov (United States)

    Su, Jing; Zhou, Feng; Lu, Da-Bing

    2013-10-01

    About 46 mammal species have been suspected as reservoir hosts for Schistosoma japonicum and therefore the track of the target parasites, in relation to definitive host species, may be of great importance in terms of theoretical and practical implications. The circadian rhythm of cercariae emergence, a genetically controlled behavior for parasites to adapt to their definitive hosts, may seem to be a perfect biological marker for S. japonicum. In this study, a late (or nocturnal) cercarial emergence pattern was observed on the parasites from one hilly region in Anhui of China, where rodents serve as reservoirs, and on the first generation of the parasites. Moreover, by using the circular statistics, the homogeneity of parasites in such trait was also demonstrated. All these provide evidence for the genetically controlled biological trait, which seems essential in the investigation of macro- or micro-dynamics of parasite transmission of interest. This is particularly true in the case of S. japonicum when multiple parasite isolates or strains are more likely to exist.

  9. High prevalence of Schistosoma japonicum and Fasciola gigantica in bovines from Northern Samar, the Philippines.

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    Catherine A Gordon

    2015-02-01

    Full Text Available The cause of zoonotic schistosomiasis in the Philippines is Schistosoma japonicum, which infects up to 46 mammalian hosts, including humans and bovines. In China, water buffaloes have been identified as major reservoir hosts for schistosomiasis japonica, contributing up to 75% of human transmission. In the Philippines, water buffaloes (carabao; Bubalus bubalis carabanesis have, historically, been considered unimportant reservoirs. We therefore revisited the possible role of bovines in schistosome transmission in the Philippines, using the recently described formalin-ethyl acetate sedimentation (FEA-SD technique and a qPCR assay to examine fecal samples from 153 bovines (both carabao and cattle from six barangays in Northern Samar. A high prevalence of S. japonicum was found using qPCR and FEA-SD in both cattle (87.50% and 77.08%, respectively and carabao (80.00% and 55.24%, respectively. The average daily egg output for each bovine was calculated at 195,000. High prevalence and infection intensity of F. gigantica was also found in the bovines by qPCR and FEA-SD (95.33% and 96.00%, respectively. The identification of bovines as major reservoir hosts for S. japonicum transmission suggests that bovine treatment and/or vaccination, as one becomes available, should be included in any future control program that aims to reduce the disease burden due to schistosomiasis in the Philippines.

  10. Diversification of Schistosoma japonicum in Mainland China revealed by mitochondrial DNA.

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    Qin Ping Zhao

    Full Text Available BACKGROUND: Schistosoma japonicum still causes severe parasitic disease in mainland China, but mainly in areas along the Yangtze River. However, the genetic diversity in populations of S. japonicum has not been well understood across its geographical distribution, and such data may provide insights into the epidemiology and possible control strategies for schistosomiasis. METHODOLOGY/PRINCIPAL FINDINGS: In this study infected Oncomelania snails were collected from areas in the middle and lower (ML reaches of the Yangtze River, including Hubei, Hunan, Anhui, Jiangxi and Jiangsu provinces, and in the upper reaches of the river, including Sichuan and Yunnan provinces in southwest (SW China. The adult parasites obtained from experimentally infected mice using isolated cercariae were sequenced individually for several fragments of mitochondrial regions, including Cytb-ND4L-ND4, 16S-12S and ND1. Populations in the ML reaches exhibited a relatively high level of diversity in nucleotides and haplotypes, whereas a low level was observed for populations in the SW, using either each single fragment or the combined sequence of the three fragments. Pairwise analyses of F-statistics (Fst revealed a significant genetic difference between populations in the ML reaches and those in the SW, with limited gene flow and no shared haplotypes in between. It is rather obvious that genetic diversity in the populations of S. japonicum was significantly correlated with the geographical distance, and the geographical separation/isolation was considered to be the major factor accounting for the observed difference between populations in the ML reaches and those in the SW in China. CONCLUSIONS: S. japonicum in mainland China exhibits a high degree of genetic diversity, with a similar pattern of genetic diversity as observed in the intermediate host snails in the same region in China.

  11. PRELIMINARY STUDY ON SCREENING OF EGG MIMOTOPE OF SCHISTOSOMA JAPONICUM%日本血吸虫模拟虫卵抗原的初步研究

    Institute of Scientific and Technical Information of China (English)

    钱旻; 白艳军; 章平; 吴自荣; 严自助

    2000-01-01

    @@ The egg of Schistosoma japonicum is a chief pathogenic factor. The preparation of egg antigen is a troublesome work, and needs a large number of experimental animals. In order to explore and develop an egg mimotope that can be used in the study of diagnostic reagents and vaccines, we employed the phage display random 15-peptides library to mimic the antigen epitope recognized by mAb 6B12 which is specific to egg antigen of Schistosoma japonicum.

  12. Immunization of pigs against infection with Schistosoma japonicum using ultraviolet-attenuated cercariae

    Energy Technology Data Exchange (ETDEWEB)

    Shi, Y.-E.; Jiang, C.-F.; Han, J.-J.; Li, Y.-L. (Tongji Medical Univ., Wuhan (China). Dept. of Parasitology); Ruppel, A. (Institute for Tropical Hygiene, Heidelberg (Germany))

    1993-06-01

    Since pigs are important in the zoonotic transmission of schistosomiasis japonica in China, a veterinary vaccine might contribute to the control of the disease in humans. Pigs were immunized with three doses each of 10 000 cercariae of Schistosoma japonicum attenuated with ultraviolet light (400 [mu]Watt.min/cm[sup 2]). The experiment was performed with portable irradiation equipment in a rural area of the Hubei Province (P.R. China). A challenge infection of 1000 untreated cercariae was given 2.5 or 6 months after the last immunization, and age-matched naive pigs were challenged as a control. Immunized pigs developed about 90% resistance against the challenge. The liver egg load of these animals was reduced by over 90%. Less than 0.01% of the immunizing cercariae developed to adult parasites and the vaccination had no apparent adverse influence on the pig's health. (Author).

  13. Delayed tail loss during the invasion of mouse skin by cercariae of Schistosoma japonicum.

    Science.gov (United States)

    Wang, Ting; Fang, Zheng-Ming; Lei, Jia-Hui; Guan, Fei; Liu, Wen-Qi; Bartlett, Ann; Whitfield, Phil; Li, Yong-Long

    2012-02-01

    A traditional assumption is that schistosome cercariae lose their tails at the onset of penetration. It has, however, recently been demonstrated that, for Schistosoma mansoni, cercarial tails were not invariably being shed as penetration took place and a high proportion of tails entered human skin under experimental conditions. This phenomenon was termed delayed tail loss (DTL). In this paper, we report that DTL also happens with S. japonicum cercariae during penetration of mouse skin. It occurred at all cercarial densities tested, from as few as 10 cercariae/2·25 cm(2) of mouse skin up to 200 cercariae. Furthermore, it was demonstrated that there was a density-dependent increase in DTL as cercarial densities increased. No such density-dependent enhancement was shown for percentage attachment over the same cercarial density range.

  14. Local Antiglycan Antibody Responses to Skin Stage and Migratory Schistosomula of Schistosoma japonicum.

    Science.gov (United States)

    Smit, Cornelis H; Kies, Christiaan L; McWilliam, Hamish E G; Meeusen, Els N T; Hokke, Cornelis H; van Diepen, Angela

    2015-10-12

    Schistosomiasis is a tropical disease affecting over 230 million people worldwide. Although effective drug treatment is available, reinfections are common, and development of immunity is slow. Most antibodies raised during schistosome infection are directed against glycans, some of which are thought to be protective. Developing schistosomula are considered most vulnerable to immune attack, and better understanding of local antibody responses raised against glycans expressed by this life stage might reveal possible glycan vaccine candidates for future vaccine research. We used antibody-secreting cell (ASC) probes to characterize local antiglycan antibody responses against migrating Schistosoma japonicum schistosomula in different tissues of rats. Analysis by shotgun Schistosoma glycan microarray resulted in the identification of antiglycan antibody response patterns that reflected the migratory pathway of schistosomula. Antibodies raised by skin lymph node (LN) ASC probes mainly targeted N-glycans with terminal mannose residues, Galβ1-4GlcNAc (LacNAc) and Galβ1-4(Fucα1-3)GlcNAc (LeX). Also, responses to antigenic and schistosome-specific glycosphingolipid (GSL) glycans containing highly fucosylated GalNAcβ1-4(GlcNAcβ1)n stretches that are believed to be present at the parasite's surface constitutively upon transformation were found. Antibody targets recognized by lung LN ASC probes were mainly N-glycans presenting GalNAcβ1-4GlcNAc (LDN) and GlcNAc motifs. Surprisingly, antibodies against highly antigenic multifucosylated motifs of GSL glycans were not observed in lung LN ASC probes, indicating that these antigens are not expressed in lung stage schistosomula or are not appropriately exposed to induce immune responses locally. The local antiglycan responses observed in this study highlight the stage- and tissue-specific expression of antigenic parasite glycans and provide insights into glycan targets possibly involved in resistance to S. japonicum infection.

  15. Screening and Primary Characterization of New Antigen Genes of Schistosoma Japonicum

    Institute of Scientific and Technical Information of China (English)

    王敏; 易新元; 李先平; 曾宪芳; 章洁

    2004-01-01

    To find Schistosoma japonicum(S.j) new antigen gene thus provide more useful vaccine candidates, the cDNA library of S. j adult worm was screened with sera of rabbits immtmized with the membrane antigens of Schistosoma japonicum hepato-portal schistosomula (SjHmAg). The positive clones were amplified by PCR and sequenced, then the sequences of clones were compared with all sequences in GenBank database using Blast process. The new clones were submitted to GenBank for accession numbers. Fifteen positive clones were obtained after three rounds of i mmunoscreening. The size of S. j cDNA fragments in positive clones ranged from 0.7 kb-3.0 kb after automatically excised with the helper phage. Sequence analysis revealed that partial sequence of clone M5 had significant homology with S.j mitochondria rnRNA, the other positive clones were new S.j genes. M2 clone sequence (GenBank accession number AF502579) was 730 bp long it had a 117 bp open reading frame (ORF). The sequence of M15 (GenBank accession number AF502582) has no transmembrane region and encodes 92 amino acids, and its protein contains a ferredoxins iron-sulfur binding region signature and two VWFC signal regions. The size of M1 ,M8, M9, M12( GenBank accession numbers: AF502578, AF502580, AF500622, AF502581 ) ranges from 402 bp to 766 bp. It concluded that the sera from rabbit immunized with SjHmAg could recognize S. j specific antigens molecules, and these antigens may induce the protective immunity against S.j infection.

  16. Spatial distribution of human Schistosoma japonicum infections in the Dongting Lake Region, China.

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    Giovanna Raso

    Full Text Available BACKGROUND: The aim of this study was to spatially model the effect of demographic, reservoir hosts and environmental factors on human Schistosoma japonicum infection prevalence in the Dongting Lake area of Hunan Province, China and to determine the potential of each indicator in targeting schistosomiasis control. METHODOLOGY/PRINCIPAL FINDINGS: Cross-sectional serological, coprological and demographic data were obtained from the 2004 nationwide periodic epidemiologic survey for Hunan Province. Environmental data were downloaded from the USGS EROS data centre. Bayesian geostatistical models were employed for spatial analysis of the infection prevalence among study participants. A total of 47,139 participants from 47 administrative villages were selected. Age, sex and occupation of residents and the presence of infected buffaloes and environmental factors, i.e. NDVI, distance to the lake and endemic type of setting, were significantly associated with S. japonicum infection prevalence. After taking into account spatial correlation, however, only demographic factors (age, sex and occupation and the presence of infected buffaloes remained significant indicators. CONCLUSIONS/SIGNIFICANCE: Long established demographic factors, as well presence of host reservoirs rather than environmental factors are driving human transmission. Findings of this work can be used for epidemiologic surveillance and for the future planning of interventions in the Dongting Lake area of Hunan Province.

  17. Molecular cloning and characterization of glutamine synthetase, a tegumental protein from Schistosoma japonicum.

    Science.gov (United States)

    Qiu, Chunhui; Hong, Yang; Cao, Yan; Wang, Fei; Fu, Zhiqiang; Shi, Yaojun; Wei, Meimei; Liu, Shengfa; Lin, Jiaojiao

    2012-12-01

    Glutamine synthetase catalyzes the synthesis of glutamine, providing nitrogen for the production of purines, pyrimidines, amino acids, and other compounds required in many pivotal cellular events. Herein, a full-length cDNA encoding Schistosoma japonicum glutamine synthetase (SjGS) was isolated from 21-day schistosomes. The entire open reading frame of SjGS contains a 1,095-bp coding region corresponding to 364 amino acids with a calculated molecular weight of 40.7 kDa. NCBIP blast shows that the putative amino acid of SjGS contains a classic β-grasp domain and a catalytic domain of glutamine synthetase. The relative mRNA expression of SjGS was evaluated in 7-, 13-, 21-, 28-, 35-, and 42-day worms of S. japonicum in the final host and higher expression at day 21, and 42 worms were observed. This protein was also detected in worm extracts using Western blot. Immunofluorescence studies indicated that the SjGS protein was mainly distributed on tegument and parenchyma in 28-day adult worms. The recombinant glutamine synthetase with a molecular weight of 45 kDa was expressed in Escherichia coli and purified in its active form. The enzyme activity of the recombinant protein was 3.30 ± 0.67 U.μg-1. The enzyme activity was highly stable over a wide range of pH (6-9) and temperature (25-40 °C) under physiological conditions. The transcription of SjGS was upregulated in praziquantel-treated worms at 2-, 4-, and 24-h posttreatment compared with the untreated control. As a first step towards the clarification of the role of glutamine synthetase in schistosome species, we have cloned and characterized cDNAs encoding SjGS in S. japonicum, and the data presented suggest that SjGS is an important molecule in the development of the schistosome.

  18. A strategy for emergency treatment of Schistosoma japonicum-infested water

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    Li Hong-Jun

    2011-11-01

    Full Text Available Abstract Background Schistosomiasis japonica, caused by contact with Schistosoma japonicum cercaria-infested water when washing, bathing or production, remains a major public-health concern in China. The purpose of the present study was to investigate the effect of a suspension concentrate of niclosamide (SCN on killing cercaria of S. japonicum that float on the water surface, and its toxicity to fish, so as to establish an emergency-treatment intervention for rapidly killing cercaria and eliminating water infectivity. Results At 30 min after spraying 100 mg/L SCN, with niclosamide dosages of 0.01, 0.02, 0.03, 0.04 g/m2, the water infectivity reduced significantly and no infectivity was found at 60 min after spraying SCN. The surface of static water was sprayed with 100 mg/L SCN, the peak concentration was found at 0 min, and the solution diffused to site with a water depth of 10 cm after 10 min. 30 min later, SCN diffused to the whole water body, and distributed evenly. After spraying 100 mg/L SCN onto the surface of the water with a volume of(3.14 × 202×50cm3, with niclosamide dosages of 0.02 g/m2, 96 h later, no death of zebra fish was observed. Conclusions By spraying 100 mg/L SCN, with a niclosamide dosage of 0.02 g/m2 onto the surface of S. japonicum-infested water, infectivity of the water can be eliminated after 30-60 min, and there is no evident toxicity to fish. This cercaria-killing method, as an emergency-treatment intervention for infested water, can be applied in those forecasting and early warning systems for schistosomiasis.

  19. Characterization and expression of a novel cystatin gene from Schistosoma japonicum.

    Science.gov (United States)

    He, Baohua; Cai, Guobin; Ni, Yonghui; Li, Ying; Zong, Hongying; He, Li

    2011-08-01

    Cystatins are a family of cysteine protease inhibitors that play a crucial role in the immune evasion from their host and in the adaptation to host defence. Here, we isolated a full-length cDNA sequence inferred to encode a novel cystatin gene from a blood fluke, Schistosoma japonicum. The cDNA, designated SjCystatin, comprised an open reading frame (ORF) of 306 bp, and encoded 101 amino acids with a predicted molecular weight of 11.3 kDa. This predicted protein shared a significant degree of sequence identity with the type I cystatin (stefin) of Schistosoma mansoni and Homo sapiens. These proteins exhibited a typical cystatin topology, including the absence of disulfide bonds and three conserved catalytic motifs, Gly at the N-terminus (Gly(6)), Gln-X-Val-X-Gly motif (Q(49)VVAG(53)) and an LP pair at the C-terminus (L(76)P(77)). The SjCystatin gene spanned 376 bp and contained three exons. The positions of two introns were conserved between the cystatin genes of trematodes and their vertebrate hosts. Reverse transcription polymerase chain reaction confirmed the transcription of SjCystatin in the egg, schistosomula and adult stages of S. japonicum. The encoding ORF region was cloned into pET-28a (+) prokaryotic expression vector. After purification, the recombinant protein SjCystatin (recSjCystatin), expressed in Escherichia coli, was used to immunize animals and produce its specific polyclonal antibody. Western blot analysis revealed that the native SjCystatin was expressed in the egg and adult stages. The enzyme activity assay of the recSjCystatin showed that it inhibited the proteolytic activity of papain. SjCystatin protein was mainly localized on the miracidium within eggs. Immunohistochemistry revealed that SjCystatin mainly localized in the epithelial cells lining the gut as well as the tegument on the surface of adult worms. The conserved genomic DNA structure among cystatin homologues of trematode and their vertebrate host emphasized the characteristics

  20. An IL-13 promoter polymorphism associated with liver fibrosis in patients with Schistosoma japonicum.

    Science.gov (United States)

    Long, Xin; Chen, Qian; Zhao, Jianping; Rafaels, Nicholas; Mathias, Priyanka; Liang, Huifang; Potee, Joseph; Campbell, Monica; Zhang, Bixiang; Gao, Li; Georas, Steve N; Vercelli, Donata; Beaty, Terri H; Ruczinski, Ingo; Mathias, Rasika; Barnes, Kathleen C; Chen, Xiaoping

    2015-01-01

    The aim of this study was to determine whether two polymorphisms in the gene encoding IL13 previously associated with Schistosoma hematobium (S. hematobium) and S. mansoni infection are associated with S. japonicum infection. Single nucleotide polymorphisms (SNPs) rs1800925 (IL13/-1112C>T) and rs20541 (IL13R130Q) were genotyped in 947 unrelated individuals (307 chronically infected, 339 late-stage with liver fibrosis, 301 uninfected controls) from a schistosomiasis-endemic area of Hubei province in China. Regression models were used to evaluate allelic and haplotypic associations with chronic and late-stage schistosomiasis adjusted for non-genetic covariates. Expression of IL-13 was measured in S. japonicun-infected liver fibrosis tissue and normal liver tissue from uninfected controls by immunohistochemistry (IHC). The role of rs1800925 in IL-13 transcription was further determined by Luciferase report assay using the recombinant PGL4.17-rs180092 plasmid. We found SNP rs1800925T was associated with late-stage schistosomiasis caused by S. japonicum but not chronic schistosomiasis (OR = 1.39, 95%CI = 1.02-1.91, p = 0.03) and uninfected controls (OR = 1.49, 95%CI = 1.03-2.13, p = 0.03). Moreover, the haplotype rs1800925T-rs20541C increased the risk of disease progression to late-stage schistosomiasis (OR = 1.46, p = 0.035), whereas haplotype rs1800925C-rs20541A showed a protective role against development of late-stage schistosomiasis (F = 0.188, OR = 0.61, p = 0.002). Furthermore, S. japonicum-induced fibrotic liver tissue had higher IL13 expression than normal liver tissue. Plasmid PGL4.17-rs1800925T showed a stronger relative luciferase activity than Plasmid PGL4.17-rs1800925C in 293FT, QSG-7701 and HL-7702 cell lines. In conclusion, the functional IL13 polymorphism, rs1800925T, previously associated with risk of schistosomiasis, also contributes to risk of late-stage schistosomiasis caused by S. japonicum.

  1. Exosome-like vesicles derived by Schistosoma japonicum adult worms mediates M1 type immune- activity of macrophage.

    Science.gov (United States)

    Wang, Lifu; Li, Zhitao; Shen, Jia; Liu, Zhen; Liang, Jinyi; Wu, Xiaoying; Sun, Xi; Wu, Zhongdao

    2015-05-01

    Exosomes are 30-100-nm membrane vesicles of endocytic origin that are released into the extracellular space upon fusion of the multi-vesicular bodies (MVB) with the plasma membrane, while initial studies described that the role of exosomes was a reticulocyte cargo-disposal mechanism allowing remodeling of the plasma membrane during the maturation of reticulocytes to erythrocytes. Recent studies indicate that exosomes are secreted by most cells and pathogens and play an important role in intercellular signaling and exert regulatory function by carrying bioactive molecules. As numerous pathogens, adult worm of Schistosoma japonicum (S. japonicum) reside in mesenteric veins of definitive host including man and mammal animals. It was reported that the worms or the eggs also have specialized secretion systems to export effector proteins or other molecules into host target cells. However, the mechanisms involved remained unclear. This study investigated the isolation of the exosome-like vesicles secreted by S. japonicum adult worms and its immune activity on microphage in vitro. In this report, we identified exosome-based secretion as a new mechanism for protein secretion by S. japonicum. Electron microscopy tomography revealed the previously unidentified ultrastructural detail of exosome-like vesicles with high resolution; they were found to be typical spherical shape and to have a diverse population that varies in size of 30-100 nm. Exosome-like vesicles isolated from S. japonicum contained a significantly different protein compared with debris pelleted and the apoptosis body. We also demonstrate that macrophages were preferentially differentiated into the M1 subtype while being treated with S. japonicum exosome-like vesicles. This study reveals there are exosome-like vesicles derived by S. japonicum adult worms, and the exosome-like vesicles can mediate M1-type immune- activity of macrophage.

  2. Silver-enhanced colloidal gold metalloimmunoassay for Schistosoma japonicum antibody detection.

    Science.gov (United States)

    Chu, Xia; Xiang, Zhi-Feng; Fu, Xin; Wang, Shi-Ping; Shen, Guo-Li; Yu, Ru-Qin

    2005-06-01

    A silver-enhanced colloidal gold metalloimmunoassay has been proposed for the determination of Schistosoma japonicum antibody (SjAb) in rabbit serum. The adult worm antigen of S. japonicum (SjAg) was adsorbed passively on the walls of a polystyrene microwell and then reacted with the desired SjAb. The colloidal gold-labeled goat anti-rabbit IgG secondary antibody was adsorbed on the walls of the polystyrene microwells through the reaction with SjAb, followed by the silver enhancement process, dissolution of silver metal atoms in an acidic solution, and determination of dissolved silver ions by anodic stripping voltammetry (ASV) at a glassy-carbon electrode. Assay conditions were optimized, including the reaction time of SjAg with SjAb, the interaction of SjAb with the colloidal gold-labeled secondary antibody, the dilution ratio of the colloidal gold-labeled secondary antibody and the silver enhancement time. The integration of the anodic stripping peak current depended linearly on the SjAb logarithmic concentration over the range of 6.4 ng/ml to 100 microg/ml. A detection limit as low as 3.0 ng/ml SjAb was achieved, which was better than the piezoelectric body acoustic wave sensor (detection limit of 7.2 microg/ml) and the renewable amperometric immunosensor (detection limit of 0.36 microg/ml). Rabbit serum samples with various degrees of infection were analyzed, and the results demonstrate that the proposed method meets the requirements of clinical analysis.

  3. A deep analysis of the small non-coding RNA population in Schistosoma japonicum eggs.

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    Pengfei Cai

    Full Text Available BACKGROUND: Schistosoma japonicum is a parasitic flatworm that causes zoonotic schistosomiasis. The typical outcome of schistosomiasis is hepatic granuloma and fibrosis, which is primarily induced by soluble egg-derived antigens. Although schistosomal eggs represent an important pathogenic stage to the host, the biology of this critical stage is largely unknown. We previously investigated the expression profiles of sncRNAs during different developmental stages of this parasite. However, using small RNA extracted from egg-deposited liver tissues generated limited information about sncRNAs in eggs. Here, we characterized the complete small RNAome in this stage of the parasite after optimization of RNA purification. METHODOLOGY AND PRINCIPAL FINDINGS: A library, SjE, was constructed with the small RNA extracted from S. japonicum eggs and subjected to high-throughput sequencing. The data were depicted by comprehensive bioinformatic analysis to explore the expression features of sncRNAs in the egg stage. MicroRNAs accounted for about one quarter of the total small RNA population in this stage, with a strongly biased expression pattern of certain miRNA family members. Sja-miR-71, sja-miR-71-5p, and sja-miR-36-3p were suggested to play important roles in embryo development. A panel of transfer RNA fragments (tRFs precisely processed from the 5' end of mature tRNAs was identified for the first time, which represented a strong egg stage-biased expression. The tRNA-Ala derived small RNAs were the most highly expressed Sj-tRFs in eggs. Further, the expression of siRNAs from 29 types of well-defined transposable elements (TEs was observed to be relatively stable among different developmental stages. CONCLUSIONS AND SIGNIFICANCE: In this study, we characterized the sncRNA profile in the egg stage of S. japonicum. Featured expression of sncRNAs, especially the tRNA-derived small RNAs, was identified, which was further compared with that of other developmental

  4. Evaluation of immunoassays for the diagnosis of Schistosoma japonicum infection using archived sera.

    Directory of Open Access Journals (Sweden)

    Jing Xu

    Full Text Available BACKGROUND: With a national program initiated recently to reduce transmission of Schistosoma japonicum in the People's Republic of China (P.R. China, there is an urgent need for accessible, quality-assured diagnostics for case detection, surveillance, and program monitoring of chemotherapy efficacy and other control interventions in areas of low endemicity. We compared the performance of nine immunodiagnostic tests developed in P.R. China for detection of antibodies against S. japonicum and established their priority for further assessment in field settings. METHODOLOGY/PRINCIPAL FINDINGS: Using the Kato-Katz technique as the reference standard, 240 well-characterized archived serum specimens (100 positive and 140 negative were evaluated in nine immunological tests developed in P.R. China. The enzyme-linked immunoelectrotransfer blot assay (EITB, which uses an adult worm extract of S. japonicum, supplied by the Center of Disease Control and Prevention, USA, was also evaluated. The sensitivity and specificity of each test were determined and the reproducibility of each test was assessed by evaluating operator-to-operator and run-to-run variation. In addition the simplicity of use for the end-user was evaluated. All tests showed good sensitivities ranging from 92.0% (95% confidence interval (CI: 86.7-97.3% to 98.0% (95% CI: 95.3-100.0%. The test specificities varied from 70.0% (95% CI: 62.4-77.6% to 97.1% (95% CI: 94.4-99.9%. All tests showed excellent reproducibility with a discordant rate in the range of 0-10.0% for operator-to-operator variation and run-to-run variation. All tests, except one magnetic particle-based enzyme-linked immunosorbent assay, were found to be easy to use, especially the dot immunogold filtration assays. CONCLUSIONS/SIGNIFICANCE: Most evaluated tests had acceptable performance characteristics and could make an impact on the schistosomiasis control programs in P.R. China. Three tests with the highest sensitivity

  5. Evaluation of Immunoassays for the Diagnosis of Schistosoma japonicum Infection Using Archived Sera

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    Xu, Jing; Peeling, Rosanna W.; Chen, Jia-Xu; Wu, Xiao-Hua; Wu, Zhong-Dao; Wang, Shi-Ping; Feng, Ting; Chen, Shao-Hong; Li, Hao; Guo, Jia-Gang; Zhou, Xiao-Nong

    2011-01-01

    Background With a national program initiated recently to reduce transmission of Schistosoma japonicum in the People's Republic of China (P.R. China), there is an urgent need for accessible, quality-assured diagnostics for case detection, surveillance, and program monitoring of chemotherapy efficacy and other control interventions in areas of low endemicity. We compared the performance of nine immunodiagnostic tests developed in P.R. China for detection of antibodies against S. japonicum and established their priority for further assessment in field settings. Methodology/Principal Findings Using the Kato-Katz technique as the reference standard, 240 well-characterized archived serum specimens (100 positive and 140 negative) were evaluated in nine immunological tests developed in P.R. China. The enzyme-linked immunoelectrotransfer blot assay (EITB), which uses an adult worm extract of S. japonicum, supplied by the Center of Disease Control and Prevention, USA, was also evaluated. The sensitivity and specificity of each test were determined and the reproducibility of each test was assessed by evaluating operator-to-operator and run-to-run variation. In addition the simplicity of use for the end-user was evaluated. All tests showed good sensitivities ranging from 92.0% (95% confidence interval (CI): 86.7–97.3%) to 98.0% (95% CI: 95.3–100.0%). The test specificities varied from 70.0% (95% CI: 62.4–77.6%) to 97.1% (95% CI: 94.4–99.9%). All tests showed excellent reproducibility with a discordant rate in the range of 0–10.0% for operator-to-operator variation and run-to-run variation. All tests, except one magnetic particle-based enzyme-linked immunosorbent assay, were found to be easy to use, especially the dot immunogold filtration assays. Conclusions/Significance Most evaluated tests had acceptable performance characteristics and could make an impact on the schistosomiasis control programs in P.R. China. Three tests with the highest sensitivity, specificity

  6. Mefloquine in combination with hemin causes severe damage to adult Schistosoma japonicum in vitro.

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    Xiao, Shu-hua; Qiao, Chunhua; Xue, Jian; Wang, Lili

    2014-03-01

    In order to explore the interaction of mefloquine with hemin against adult Schistosoma japonicum in vitro, the 50% and 95% lethal concentration (LC50 and LC95) of mefloquine and hemin against schistosomes, some factors, such as other iron providing agents, iron chelaters, zinc protoporphyrin-IX, and biological relevant reductants, that might impact on antischistosomal activity induced by interaction of mefloquine with hemin, and preliminary analysis of chemical interaction of both compounds were undertaken. The LC50 and LC95 of mefloquine and hemin alone against schistosomes were determined to be 6.5μg/ml and 7.8μg/ml as well as 232μg/ml and 355μg/ml, respectively. The LC50 and LC95 of mefloquine in the presence of hemin 100μg/ml was 0.24μg/ml and 0.59μg/ml, respectively. On the other hand the LC50 and LC95 of hemin in the presence of mefloquine 1μg/ml was 23.2μg/ml and 77.2μg/ml, respectively. Meanwhile, mefloquine/hemin combinations showed potential synergistic effects against adult S. japonicum, with combination index (CI) values mefloquine exhibited no toxic effect against schistosomes. On the other hand, addition of iron chelators (deferiprone, desferrioxamine mesylate, or 2,2'-bipyridine) to the medium containing mefloquine-hemin resulted in no protective effect on the worms. Furthermore, biological reductants like glutathione, vitamine C or cysteine showed no apparent worm protection effect from toxic mefloquine-hemin even at higher concentrations (242.3-614.6μg/ml, i.e., 6.4-17.8-fold higher than the concentration of hemin). Chemical interaction of mefloquine with hemin was studied in 40% DMSO-Tris buffer solution. Both UV-Vis spectrum and mass spectrum demonstrated the strong interaction of mefloquine with hemin, which resulted in a reduction of hemin color and emergence of an adduct formed by mefloquine and hemin. The results confirm that mefloquine combined with hemin exhibits potential synergistic effect against adult S. japonicum in vitro.

  7. Identification and characterization of microRNAs and endogenous siRNAs in Schistosoma japonicum

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    Wang Heng

    2010-01-01

    Full Text Available Abstract Background Small endogenous non-coding RNAs (sncRNAs such as small interfering RNA (siRNA, microRNA and other small RNA transcripts are derived from distinct loci in the genome and play critical roles in RNA-mediated gene silencing mechanisms in plants and metazoa. They are approximately 22 nucleotides long; regulate mRNA stability through perfect or imperfect match to the targets. The biological activities of sncRNAs have been related to many biological events, from resistance to microbe infections to cellular differentiation. The development of the zoonotic parasite Schistosoma japonicum parasite includes multiple steps of morphological alterations and biological differentiations, which provide a unique model for studies on the functions of small RNAs. Characterization of the genome-wide transcription of the sncRNAs will be a major step in understanding of the parasite biology. The objective of this study is to investigate the transcriptional profile and potential function of the small non-coding RNAs in the development of S. japanicum. Results The endogenous siRNAs were found mainly derived from transposable elements (TE or transposons and the natural antisense transcripts (NAT. In contrast to other organisms, the TE-derived siRNAs in S. japonicum were more predominant than other sncRNAs including microRNAs (miRNAs. Further, there were distinct length and 3'end variations in the sncRNAs, which were associated with the developmental differentiation of the parasite. Among the identified miRNA transcripts, there were 38 unique to S. japonicum and 16 that belonged to 13 miRNA families are common to other metazoan lineages. These miRNAs were either ubiquitously expressed, or they exhibited specific expression patterns related to the developmental stages or sex. Genes that encoded miRNAs are mainly located in clusters within the genome of S. japonicum. However, genes within one cluster could be differentially transcribed, which suggested

  8. Schistosoma japonicum:Isolation and Identification of Peptides Mimicking Ferritin Epitopes from Phage Display Library

    Institute of Scientific and Technical Information of China (English)

    Lian-Fei TANG; Xin-Yuan YI; Xian-Fang ZENG; Lin-Qian WANG; Shun-Ke ZHANG

    2004-01-01

    In an attempt to isolate and identify the antigenic epitopes on ferritin of Schistosoma japonicum(SjFer)and to test their protective potentiality against Schistosomajaponicum(S.j),polyclonal antisera against SjFer was prepared to screen a 12-mer random peptide library.Three rounds of biopanning were performed and resulted in an enrichment.Six peptides selected randomly from the third elute were all found to be positive by evaluating the binding to anti-SjFer sera by ELISA and Western blotting.Three amino acid sequences were deduced from the six phage clones by sequencing.When they were used to vaccinate mice,the three peptides could induce significant reduction in adult worms(26.7% ,20.4%,and 25.9%)as well as in liver eggs per gram(LEPG)(40.0%,38.2%,and 40.8%).This result showed that three mimotopes on SjFer were obtained and they could induce significant protective efficacy against S.j.

  9. Chronic Schistosoma japonicum infection reduces immune response to vaccine against hepatitis B in mice.

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    Lin Chen

    Full Text Available BACKGROUND: Hepatitis B and schistosomiasis are most prevalent in Africa and Asia, and co-infections of both are frequent in these areas. The immunomodulation reported to be induced by schistosome infections might restrict immune control of hepatitis B virus (HBV leading to more severe viral infection. Vaccination is the most effective measure to control and prevent HBV infection, but there is evidence for a reduced immune response to the vaccine in patients with chronic schistosomiasis japonica. METHODOLOGY/PRINCIPAL FINDINGS: In this paper, we demonstrate in a mouse model that a chronic Schistosoma japonicum infection can inhibit the immune response to hepatitis B vaccine (HBV vaccine and lead to lower production of anti-HBs antibodies, interferon-γ (IFN-γ and interleukin-2 (IL-2. After deworming with Praziquantel (PZQ, the level of anti-HBs antibodies gradually increased and the Th2-biased profile slowly tapered. At 16 weeks after deworming, the levels of anti-HBs antibodies and Th1/Th2 cytokines returned to the normal levels. CONCLUSIONS/SIGNIFICANCE: The results suggest that the preexisting Th2-dominated immune profile in the host infected with the parasite may down-regulate levels of anti-HBs antibodies and Th1 cytokines. To improve the efficacy of HBV vaccination in schistosome infected humans it may be valuable to treat them with praziquantel (PZQ some time prior to HBV vaccination.

  10. BEBERAPA FAKTOR YANG BERHUBUNGAN DENGAN PENULARAN Schistosoma japonicum DI DATARAN TINGGI NAPU KABUPATEN POSO SULAWESI TENGAH

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    Rosmini Rosmini

    2012-07-01

    Full Text Available Abstract. The transmission of schistosomiasis is related with human behavior or habit factors in the schistosomiasis endemic area. The Purpose of this study was to identify several factors in relationship with schistosomiasis in Napu Highland. The Study was observational with cross sectional design. Data collection were coducted stool survey and interviewes in among 196 respondents using simple random sampling. Stool samples were examined by the Kato-Katz method and interviewed was used  questionnaire to identify several factors in realtionship with schistosomiasis transmission. Out of 6 community behaviours suspected to contribute to infection of schistosomiasis, 5 were related with schistosomiasis, i. e. the habit of taking a bath and washing in the river (p=<0.001, always going to the focus area (p=0.04,utilizing water resources from well (p=0.001, washing legs or hand in the river(p=<0.001 and swimming (p=0.007. Schistosomiasis infection was related to the community behaviour of bathing and washing in the river, always going to the focus area, utilizing water resources from well, washing legs or hand in the river and swimming.Key word: Schistosomiasis, Schistosoma japonicum, behaviour, Napu Highland.Central Sulawesi

  11. Expression of Recombinant Baculovirus Carrying Schistosoma japonicum 26 ku GST in Mammalian Cells

    Institute of Scientific and Technical Information of China (English)

    YU Guangqing; SONG Jianhua; LIU Wenqi; LONG Xiaochun; MO Hongmei; LI Yonglong; CHEN Xinwen

    2006-01-01

    In order to construct recombinant baculovirus carrying Schistosoma japonicum 26 ku glutathione S-transferase gene (Sj26), and observe the expression of Sj26 in mammalian cells, the Sj26 gene was amplified with plasmid pGEX-3X as template by PCR, and then recombined into Tvector for sequencing. Sj26 gene was inserted into the downstream of CMV promoter of donor plasmid pFBDGC, and the recombinant donor plasmid pFBDGC-Sj26 transformed into DH10Bac,then the recombinant bacmid AcCMVSj26 was isolated and transfected into Sf9 cells. The recombinant baculovirus was harvested and final titer of vAcCMVSj26 was measured. BHK cells were transducted with recombinant baculovirus in vitro. By using Western blot, the expression of 26 ku glutathione S-transferase (GST) was detected. The results showed that after enzyme digestion and sequencing, the donor plasmid was successfully constructed. PCR confirmed that pFBDGC-Sj26 and Bacmid homologous recombination occurred in E. coli. After transfection of Sf9 cells with recombinant Bacmid, recombinant baculovirus was replicated in Sf9 cells and expressed green fluorescent protein. PCR further revealed recombinant baculovirus contained Sj26. The titer of the harvested baculovirus was 1.24 × 108. Western blot demonstrated that recombinant baculovirus could express 26 ku GST in BHK cells. It was concluded that Sj26 recombinant baculovirus was successfully constructed, and the 26 ku GST was expressed in mammalian cells.

  12. Construction of a recombinant plasmid pGEX-Sj26GST of Schistosoma japonicum and expression in Escherichia coli BL21( DE3)

    Institute of Scientific and Technical Information of China (English)

    张丽

    2014-01-01

    Objective To construct and express a recombinant plasmid pGEX-Sj26GST of Schistosoma japonicum(Sj)in Escherichia coli(E.coli)BL21(DE3).Methods Total RNA was extracted from Sj adult worms by RNeasy Mini kit,26 kilodalton glutathione-S-transferases of Schistosoma japonicum(Sj26GST)antigen gene was amplified by real-time PCR(RT-PCR)from the total RNA,then cloned into a prokaryotic expression plasmid pGEX-1λT and transformed into E.coli BL21(DE3)to construct pGEX-Sj26GST;BL21(pGEX-

  13. Cloning and expression in Escherichia coli of a new gene of Schistosoma japonicum encoding casein kinase Ⅱ beta subunit

    Institute of Scientific and Technical Information of China (English)

    彭寨玉; 余新炳; 吴忠道; 徐劲; 吴德; 李孜

    2004-01-01

    Background Nowadays it is now a focus topic in schistosomiasis research to find ideal vaccine candidates and new drug targets for developing anti-schistosomiasis vaccine. We cloned a new gene, casein kinase Ⅱ beta subunit, of Schistosoma japonicum (S. japonicum) and express it in Escherichia coli (E.coli).Methods The ESTs obtained in our laboratory were analyzed by homologous searching, and a new gene was recognized. The full-length cDNA of the new gene was obtained by joining the 3'RACE PCR fragment and the EST clone. To express the new gene, the cDNA was cloned into pGEX-4T-1 vector and then transformed into E.coli JM109. The recombinant protein was analyzed by SDS-PAGE and Western-blot. Results A 908 bp cDNA was isolated from S. japonicum and identified to be casein kinase Ⅱ beta subunit gene by sequence analysis. The open reading frame of the gene encodes a protein of 217 amino acids exhibiting 75.8%, 75.8%, 73.9%, 68.2%, 51.6% identity to the amino acids sequence of the corresponding genes of Homo sapiens (H. sapiens), Xenopus laevi (X. laevi), Drosophila melanogaster (D. melanogaster), Caenorhabditis elegan (C. elegan), and Schizosaccharomyces pombe (S. promber) respectively. The predicted molecular weight of the protein was 24.921 kDa. The new cDNA sequence had been submitted to GenBank, and its accession number is AY241391. This cDNA was subcloned into the pGEX-4T-1 vector and expressed in E.coli JM109.The recombinant protein could be recognized by the S. japonicum infected rabbit serum. Conclusion The full-length cDNA sequences encoding S. japonicum casein kinase Ⅱ beta subunit were firstly sequenced, cloned, and expressed in E.coli.

  14. Upregulated Expression of Cytotoxicity-Related Genes in IFN-γ Knockout Mice with Schistosoma japonicum Infection

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    Xiaotang Du

    2011-01-01

    Full Text Available It is well accepted that IFN-γ is important to the development of acquired resistance against murine schistosomiasis. However, the in vivo role of this immunoregulatory cytokine in helminth infection needs to be further investigated. In this study, parasite burden and host immune response were observed in IFN-γ knockout mice (IFNg KO infected with Schistosoma japonicum for 6 weeks. The results suggested that deficiency in IFN-γ led to decreased egg burden in mice, with low schistosome-specific IgG antibody response and enhanced activation of T cells during acute infection. Microarray and qRT-PCR data analyses showed significant upregulation of some cytotoxicity-related genes, including those from the granzyme family, tumor necrosis factor, Fas Ligand, and chemokines, in the spleen cells of IFNg KO mice. Furthermore, CD8+ cells instead of NK cells of IFNg KO mice exhibited increased transcription of cytotoxic genes compared with WT mice. Additionally, Schistosoma japonicum-specific egg antigen immunization also could activate CD8+ T cells to upregulate the expression of cytotoxic genes in IFNg KO mice. Our data suggest that IFN-γ is not always a positive regulator of immune responses. In certain situations, the disruption of IFN-γ signaling may up-regulate the cytotoxic T-cell-mediated immune responses to the parasite.

  15. Effect of photoperiod change on chronobiology of cercarial emergence of Schistosoma japonicum derived from hilly and marshy regions of China.

    Science.gov (United States)

    Wang, Su-Rong; Zhu, Yuan-Jian; Ge, Qing-Peng; Yang, Meng-Jia; Huang, Ji-Lei; Huang, Wen-Qiao; Zhuge, Hong-Xiang; Lu, Da-Bing

    2015-12-01

    The chronobiology of cercarial emergence appeared to be a genetically controlled behavior, adapted to definitive host species, for schistosome. However, a few physiological and ecological factors, for example the change of photoperiod, were reported to affect the rhythmic emergence of cercariae. Therefore, the effect of photoperiod change on cercarial emergence of two Schistosoma japonicum isolates, the hilly and the marshland, was investigated. Four shedding experiments each under a different photoperiod were conducted. Under a natural photoperiod, two distinct shedding modes, one from the hilly region and one from the marshland, were observed. Under a reversed photoperiod, the regular pattern (i.e. under a natural photoperiod) of S. japonicum cercarial emergence was reversed for the marshland isolate and disappeared for the hilly isolate. With an input of a 2 h darkness from 7am to 9am, the cercarial emergence peak were delayed for the two isolates; whereas with an input of a 2 h darkness from 5pm to 7pm, neither effect on the cercarial emergence rhythm was observed. The total cercariae emerged for both parasite isolates varied with a different photoperiod. The results indicate that the change of photoperiod could affect the chronobiology of S japonicum cercarial emergence.

  16. Combined TLR7/8 and TLR9 ligands potentiate the activity of a Schistosoma japonicum DNA vaccine.

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    Xuefeng Wang

    Full Text Available BACKGROUND: Toll-like receptor (TLR ligands have been explored as vaccine adjuvants for tumor and virus immunotherapy, but few TLR ligands affecting schistosoma vaccines have been characterized. Previously, we developed a partially protective DNA vaccine encoding the 26-kDa glutathione S-transferase of Schistosoma japonicum (pVAX1-Sj26GST. METHODOLOGY/PRINCIPAL FINDINGS: In this study, we evaluated a TLR7/8 ligand (R848 and a TLR9 ligand (CpG oligodeoxynucleotides, or CpG as adjuvants for pVAX1-Sj26GST and assessed their effects on the immune system and protection against S. japonicum. We show that combining CpG and R848 with pVAX1-Sj26GST immunization significantly increases splenocyte proliferation and IgG and IgG2a levels, decreases CD4(+CD25(+Foxp3(+ regulatory T cells (Treg frequency in vivo, and enhances protection against S. japonicum. CpG and R848 inhibited Treg-mediated immunosuppression, upregulated the production of interferon (IFN-γ, tumor necrosis factor (TNF-α, interleukin (IL-4, IL-10, IL-2, and IL-6, and decreased Foxp3 expression in vitro, which may contribute to prevent Treg suppression and conversion during vaccination and allow expansion of antigen-specific T cells against pathogens. CONCLUSIONS: Our data shows that selective TLR ligands can increase the protective efficacy of DNA vaccines against schistosomiasis, potentially through combined antagonism of Treg-mediated immunosuppression and conversion.

  17. An "in-depth" description of the small non-coding RNA population of Schistosoma japonicum schistosomulum.

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    Zhangxun Wang

    Full Text Available Parasitic flatworms of the genus Schistosoma are the causative agents of schistosomiasis, which afflicts more than 200 million people yearly in tropical regions of South America, Asia and Africa. A promising approach to the control of this and many other diseases involves the application of our understanding of small non-coding RNA function to the design of safe and effective means of treatment. In a previous study, we identified five conserved miRNAs from the adult stage of Schistosoma japonicum. Here, we applied Illumina Solexa high-throughput sequencing methods (deep sequencing to investigate the small RNAs expressed in S. japonicum schistosomulum (3 weeks post-infection. This has allowed us to examine over four million sequence reads including both frequently and infrequently represented members of the RNA population. Thus we have identified 20 conserved miRNA families that have orthologs in well-studied model organisms and 16 miRNA that appear to be specific to Schistosoma. We have also observed minor amounts of heterogeneity in both 3' and 5' terminal positions of some miRNA as well as RNA fragments resulting from the processing of miRNA precursor. An investigation of the genomic arrangement of the 36 identified miRNA revealed that seven were tightly linked in two clusters. We also identified members of the small RNA population whose structure indicates that they are part of an endogenously derived RNA silencing pathway, as evidenced by their extensive complementarities with retrotransposon and retrovirus-related Pol polyprotein from transposon.

  18. Immunodiagnostic efficacy of detection of Schistosoma japonicum human infections in China:a meta analysis

    Institute of Scientific and Technical Information of China (English)

    Wei Wang; Youzi Li; Hongjun Li; Yuntian Xing; Guoli Qu; Jianrong Dai; Yousheng Liang

    2012-01-01

    Objective: To assess the diagnostic efficacy of the currently most widely used indirect hemagglutination assay (IHA) and enzyme-linked immunosorbent assay (ELISA) for detection of Schistosoma japonicum human infections. Methods:A comprehensive search was undertaken from China National Knowledge Infrastructure, Wanfang Database, VIP Database, PubMed, Cochrane Library, Science Citation Index Expanded, Proquest, and the inclusion and exclusion criteria were strictly settled. The funnel plot was used to assess the publication bias, Cochran’s Q test was employed to measure the homogeneity between studies, a summary receiver operating characteristic (SROC) curve was used to compare the diagnostic accuracy between the IHA and ELISA qualitatively by means of the Weighted Least Square method, the Ordinary Least Square method and the Robust regression method, and the diagnostic odds ratio (DOR) was drawn to compare the accuracy quantitatively. Results:Out of 785 publications, 19 papers were eventually selected for analysis. Literature quality assessment indicated that minor publication bias existed in studies pertaining IHA test, but no bias was found in literatures regarding ELISA test. The heterogeneity test showed a heterogeneity between studies was present (χ2=466.07 and 34.67, both P values<0.0001). The areas under the SROC curves of IHA were all higher than that of ELISA test using the three methods (Weighted Least Square method:0.766 vs. 0.695, Ordinary Least Square method: 0.826 vs. 0.741, Robust regression: 0.815 vs. 0.715). The TPR*values for IHA and ELISA were 0.710, 0.759, 0.749, and 0.650, 0.686 and 0.666, respectively, and OR values were 5.997, 9.937, 8.893, and 3.432, 4.784 and 3.959, respectively. The DOR of IHA was 9.41 (95%CI:4.88-18.18), and 4.78 (95%CI:3.21-7.13) for ELISA. Conclusions:All above results revealed that the diagnostic performance of IHA is better than that of ELISA. However, taking into account their unsatisfactory diagnostic value in

  19. Cloning, expression and characterisation of a type II cystatin from Schistosoma japonicum, which could regulate macrophage activation.

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    Yang, Xiao; Liu, Ju; Yue, Yuan; Chen, Wei; Song, Man; Zhan, Ximei; Wu, Zhongkai

    2014-11-01

    Cystatin play an important role in parasite immune evasion. It is involved in many immune responses processes regulations such as inhibiting antigen presentation, modifying cytokines production and macrophage polarization. In recent years, more and more cystatins were used in treating some inflammatory diseases such as asthma and inflammation bowel diseases; however, cystatins from Schistosoma japonicum were rarely studied. In the present study, we have cloned a cystatin from the adult stage of Schistosoma japonicum, named as SjCystatin, and its sequence shares conserved domains with other type II family cystatins. It was further verified by enzyme inhibition assays. SjCystatin retained its inhibitory activity under a wide range of pH values and temperatures, can maintain its inhibitory activity at pH 6.5-7.5 and 37 °C, respectively. Then, we investigated the effects of SjCystatin on the lipopolysaccharide (LPS)-induced activated RAW264.7. Results showed that SjCystatin inhibit LPS-induced nitric oxide production in a dose-dependent manner. LPS-induced TNF-α and IL-6 production began to be inhibited at least 6 h after SjCystatin stimulation. SjCystatin significantly increased IL-10 production at 6 h after stimulation and its effect on IL-10 production diminished quickly. These results imply that SjCystatin can induce M2 macrophage polarization and can be expected to serve as a potential drug source for the medication of inflammatory disorders like other cystatins.

  20. iTRAQ-based comparative proteomic analysis of excretory-secretory proteins of schistosomula and adult worms of Schistosoma japonicum.

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    Cao, Xiaodan; Fu, Zhiqiang; Zhang, Min; Han, Yanhui; Han, Hongxiao; Han, Qian; Lu, Ke; Hong, Yang; Lin, Jiaojiao

    2016-04-14

    Schistosomiasis remains a serious public health problem with 200 million people infected and 779 million people at risk worldwide. The schistosomulum and adult worm are two stages of the complex lifecycle of Schistosoma japonicum and excretory/secretory proteins (ESPs) play a major role in host-parasite interactions. In this study, iTRAQ-coupled LC-MS/MS was used to investigate the proteome of ESPs obtained from schistosomula and adult worms of S. japonicum, and 298 differential ESPs were identified. Bioinformatics analysis of differential ESPs in the two developmental stages showed that 161 ESPs upregulated in schistosomula were associated with stress responses, carbohydrate metabolism and protein degradation, whereas ESPs upregulated in adult worms were mainly related to immunoregulation and purine metabolism. Recombinant heat shock protein 70 (HSP70) and thioredoxin peroxidase (TPx), two differential proteins identified in this study, were expressed. Further studies showed that rSjHSP70 and rSjTPx stimulated macrophages expressing high levels of the anti-inflammatory factors TGF-β, IL-10 and Arg-1, and suppressed the expression of the pro-inflammatory cytokines TNF-α, IL-1β, IL-6 and iNOS in LPS-induced macrophages. This study provides new insights into the survival and development of schistosomes in the final host and helps identify vaccine candidates or new diagnostic reagents for schistosomiasis.

  1. Multiple vaccinations with UV- attenuated cercariae in pig enhance protective immunity against Schistosoma japonicum infection as compared to single vaccination

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    Zhang Donghui

    2011-06-01

    Full Text Available Abstract Background Schistosomiasis japonica is a major public health problem in the endemic areas of China, the Philippines, and Indonesia. To date, a vaccine has not been developed against this disease but immunization with UV-attenuated cercariae can induce a high level of protective immunity in Landrace/Yorkshire/Duroc crossbred pigs. To compare the efficacy of a single vaccination and multiple vaccinations with UV-attenuated Schistosoma japonicum cercariae, two groups of pigs received either one or three exposures to 10,000 cercariae attenuated with 400 μw UV. Results Pigs with a single immunization had a 59.33% reduction in adult worm burden, a 89.87% reduction in hepatic eggs and a 86.27% reduction in fecal eggs at eight weeks post-challenge (P P Conclusion The high levels of protection against S. japonicum infection can be achieved with a UV-attenuated vaccine in pigs, and that three vaccinations were possibly more effective than a single vaccination. Moreover, triple vaccinations evoked a more vigorous IFN-γ response and a stronger antibody-mediated response, especially an increase in the levels of IgG2 antibodies.

  2. Single- or mixed-sex Schistosoma japonicum infections of intermediate host snails in hilly areas of Anhui, China.

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    Shi, Hui-Ping; Lu, Da-Bing; Shen, Lei; Shi, Tan; Gu, Jian

    2014-02-01

    Schistosomiasis japonicum is one of the most serious communicable diseases, and the transmission of the parasite is dependent of its complex life cycle on which many factors can have an impact. Multiple infections comprising both male and female schistosome within snail intermediate hosts, for example, would facilitate parasite transmission. However, no research on Schistosoma japonicum communities in field-collected Oncomelania hupensis hupensis in relation to schistosome sex has been reported. Therefore, snail survey was performed in a hilly region of Anhui, China, and single- or mixed-sex schistosome infections of snails were detected with final host mouse infection. A total of 8,563 snails were sampled in the field, and 67 were identified with schistosome infections. Of these infected snails, 46 were selected for final host infection. From this, 21 snails were infected with female schistosome, 23 with males and 2 with both males and females. More worms were recovered for snails with mixed-sex infections than with single-sex infection and for snails with male schistosome infection than with female infection (Psnails was significantly higher than would be expected if randomly distributed (Psnails was nearly equal and up to 95.65 % (44/46) of infected snails were single-sex infection. Schistosome infections in snails collected from the hilly area of Anhui Province were not randomly distributed but over-dispersed.

  3. Variable maturation and oviposition by female Schistosoma japonicum in mice: the effects of irradiation of the host prior to infection

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    Cheever, A.W.; Duvall, R.H.

    1987-11-01

    The maturation of female Schistosoma japonicum was found to vary greatly within each of two Philippine strains of this parasite and some females did not contain uterine eggs 7 to 15 weeks after infection while others contained numerous eggs before the fifth week of infection. It was found that female worms containing less than 20 uterine eggs contributed little to the accumulation of eggs in the tissues of infected mice. Such worms also generally appeared to be immature. The variable rate of maturation of worms is likely to have profound effects on the immune reactions of mice as well as on the pathologic response to infection. Systematic delay in oviposition was serendipitously found in worms from mice which had been irradiated for other purposes prior to exposure to S. japonicum, and from the fourth to the sixth week after infection egg production by worms in irradiated mice lagged well behind that in intact mice. Seven to 10 weeks after infection these worms were laying normal numbers of eggs, as judged by egg passage per worm pair in the feces and the accumulation of eggs in the tissues. S. mansoni developed normally in irradiated mice.

  4. Enhanced expression of the decoy receptor IL-13Rα2 in macrophages of Schistosoma japonicum-infected mice

    Institute of Scientific and Technical Information of China (English)

    WANG Wei; SHEN Yu-xian; LI Jing; ZHANG Shi-hai; LUO Qing-li; ZHONG Zhen-rong; JIANG Zuo-jun; SHEN Ji-long

    2009-01-01

    Background Type 2 cytokine interleukin (IL)-13 and its decoy receptor, IL-13 receptor (R)α2 appear to play a major role in tissue fibrosis of schistosomiasis and asthma. IL-13 is a key regulator of the extracellular matrix (ECM). It is known to signal to cells by binding to the IL-13Ra1, which then heterodimerizes with IL-4Rα. In contrast, IL-13Rα2 binds IL-13 with high affinity but does not signal. IL-13Rα2 is known to down-regulate granulomatous inflammation and prolong host survival in Schistosoma mansoni (S. Mansoni) infection, but little is known about the location and expression level of IL-13Ra2 in the context of S. Japonicum infection. Methods We established S. Japonicum-infected mouse models. Kinetic serum levels of IL-13Rα2 were examined with ELISA. IL-13Rα2 mRNA and protein of liver tissues were determined by PCR and immunoblotting analysis, respectively. Detection of IL-13Rα2 expression and location in macrophages was performed by TaqMan PCR and fluorescent immunocytochemistry technique, respectively. Results A marked elevation of mRNA and protein expression of IL-13Rα2 was observed in mice during S. Japonicum infection. An enhanced expression of IL-13Rg2 was further demonstrated in primary macrophages of murine schistosomiasis. Conclusions IL-13Rα2 in macrophages may be a critical contributor to pathogenesis of schistosomiasis. The data highlight the potential importance of cell signaling and antifibrotic gene therapeutics in T helper 2 cell (Th2)-mediated diseases.

  5. Comparative characterization of microRNAs in Schistosoma japonicum schistosomula from Wistar rats and BALB/c mice.

    Science.gov (United States)

    Han, Hongxiao; Peng, Jinbiao; Hong, Yang; Fu, Zhiqiang; Lu, Ke; Li, Hao; Zhu, Chuangang; Zhao, Qiuhua; Lin, Jiaojiao

    2015-07-01

    More than 40 kinds of mammals in China are known to be naturally infected with Schistosoma japonicum (S. japonicum) (Peng et al. Parasitol Res 106:967-76, 2010). Compared with permissive BALB/c mice, rats are less susceptible to S. japonicum infection and are considered to provide an unsuitable microenvironment for parasite growth and development. MicroRNAs (miRNAs), via the regulation of gene expression at the transcriptional and post-transcriptional levels, may be responsible for developmental differences between schistosomula in these two rodent hosts. Solexa deep-sequencing technology was used to identify differentially expressed miRNAs from schistosomula isolated from Wistar rats and BALB/c mice 10 days post-infection. The deep-sequencing analysis revealed that nearly 40 % of raw reads (10.37 and 10.84 million reads in schistosomula isolated from Wistar rats and BALB/c mice, respectively) can be mapped to selected mirs in miRBase or in species-specific genomes. Further analysis revealed that several miRNAs were differentially expressed in schistosomula isolated from these two rodents; 18 were downregulated (by 2-fold) (expression levels in rats compare with those in mice). Additionally, three novel miRNAs were primarily predicted and identified. Among the 41 differentially expressed miRNAs, 4 miRNAs had been identified with specific functions in schistosome development or host-parasite interaction, such as sexual maturation (sja-miR-1, sja-miR-7-5p), embryo development (sja-miR-36-3p) in schistosome, and pathogenesis of schistosomiasis (sja-bantam). Then, the target genes were mapped, filtered, and correlated with a set of genes that were differentially expressed genes in schistosomula isolated from mice and rats, which we identified in a S. japonicum oligonucleotide microarray analysis in a previous study. Gene Ontology (GO) analysis of the predicted target genes of 13 differentially expressed miRNAs revealed that they were involved in some important

  6. The nature and combination of subunits used in epitope-based Schistosoma japonicum vaccine formulations affect their efficacy

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    Liu Feng

    2010-11-01

    Full Text Available Abstract Background Schistosomiasis remains a major public health problem in endemic countries and is caused by infections with any one of three primary schistosome species. Although there are no vaccines available to date, this strategy appears feasible since natural immunity develops in individuals suffering from repeated infection during a lifetime. Since vaccinations resulting in both Th1- and Th2-type responses have been shown to contribute to protective immunity, a vaccine formulation with the capacity for stimulating multiple arms of the immune response will likely be the most effective. Previously we developed partially protective, single Th- and B cell-epitope-based peptide-DNA dual vaccines (PDDV (T3-PDDV and B3-PDDV, respectively capable of eliciting immune responses against the Schistosoma japonicum 22.6 kDa tegument antigen (Sj22.6 and a 62 kDa fragment of myosin (Sj62, respectively. Results In this study, we developed PDDV cocktails containing multiple epitopes of S. japonicum from Sj22.6, Sj62 and Sj97 antigens by predicting cytotoxic, helper, and B-cell epitopes, and evaluated vaccine potential in vivo. Results showed that mice immunized with a single-epitope PDDV elicited either Tc, Th, or B cell responses, respectively, and mice immunized with either the T3- or B3- single-epitope PDDV formulation were partially protected against infection. However, mice immunized with a multicomponent (3 PDDV components formulation elicited variable immune responses that were less immunoprotective than single-epitope PDDV formulations. Conclusions Our data show that combining these different antigens did not result in a more effective vaccine formulation when compared to each component administered individually, and further suggest that immune interference resulting from immunizations with antigenically distinct vaccine targets may be an important consideration in the development of multicomponent vaccine preparations.

  7. Utilization of ELISA using thioredoxin peroxidase-1 and tandem repeat proteins for diagnosis of Schistosoma japonicum infection among water buffaloes.

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    Jose Ma M Angeles

    Full Text Available BACKGROUND: The presence of animal reservoirs in Schistosoma japonicum infection has been a major obstacle in the control of schistosomiasis. Previous studies have proven that the inclusion of control measures on animal reservoir hosts for schistosomiasis contributed to the decrease of human cases. Animal surveillance should therefore be included to strengthen and improve the capabilities of current serological tests. METHODOLOGY/PRINCIPAL FINDINGS: Thioredoxin peroxidase-1 (SjTPx-1 and four tandem repeat proteins (Sj1TR, Sj2TR, Sj4TR, Sj7TR were initially evaluated against human sera. The previous test showed high sensitivity and specificity for antibody detection against SjTPx-1 and Sj7TR. In this study, the immunodiagnostic potential of these recombinant proteins was evaluated using enzyme-linked immunoassay on 50 water buffalo serum samples collected in Cagayan, the Philippines as compared with the soluble egg antigen (SEA. For specificity, 3 goat serum samples positive with Fasciola hepatica were used and among the antigens used, only SEA showed cross-reaction. Stool PCR targeting the S. japonicum 82 bp mitochondrial NAD 1 gene was done to confirm the true positives and served as the standard test. Twenty three samples were positive for stool PCR. SjTPx-1 and Sj1TR gave the highest sensitivity among the recombinant proteins tested for water buffalo samples with 82.61% and 78.26% respectively which were higher than that of SEA (69.57%. CONCLUSIONS/SIGNIFICANCE: These results prove that SjTPx-1 works both for humans and water buffaloes making it a good candidate antigen for zoonotic diagnosis. Sj1TR showed good results for water buffaloes and therefore can also be used as a possible candidate for detecting animal schistosome infection.

  8. Distinct genetic diversity of Oncomelania hupensis, intermediate host of Schistosoma japonicum in mainland China as revealed by ITS sequences.

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    Qin Ping Zhao

    Full Text Available BACKGROUND: Oncomelania hupensis is the unique intermediate host of Schistosoma japonicum, which causes schistosomiasis endemic in the Far East, and especially in mainland China. O. hupensis largely determines the parasite's geographical range. How O. hupensis's genetic diversity is distributed geographically in mainland China has never been well examined with DNA sequence data. METHODOLOGY/PRINCIPAL FINDINGS: In this study we investigate the genetic variation among O. hupensis from different geographical origins using the combined complete internal transcribed spacer 1 (ITS1 and ITS2 regions of nuclear ribosomal DNA. 165 O. hupensis isolates were obtained in 29 localities from 7 provinces across mainland China: lake/marshland and hill regions in Anhui, Hubei, Hunan, Jiangxi and Jiangsu provinces, located along the middle and lower reaches of Yangtze River, and mountainous regions in Sichuan and Yunnan provinces. Phylogenetic and haplotype network analyses showed distinct genetic diversity and no shared haplotypes between populations from lake/marshland regions of the middle and lower reaches of the Yangtze River and populations from mountainous regions of Sichuan and Yunnan provinces. The genetic distance between these two groups is up to 0.81 based on Fst, and branch time was estimated as 2-6 Ma. As revealed in the phylogenetic tree, snails from Sichuan and Yunnan provinces were also clustered separately. Geographical separation appears to be an important factor accounting for the diversification of the two groups of O. hupensis in mainland China, and probably for the separate clades between snails from Sichuan and Yunnan provinces. In lake/marshland and hill regions along the middle and lower reaches of the Yangtze River, three clades were identified in the phylogenetic tree, but without any obvious clustering of snails from different provinces. CONCLUSIONS: O. hupensis in mainland China may have considerable genetic diversity, and a more

  9. Selection of Immunogenic Peptide Mimics of Male Worm Origin of Schistosoma Japonicum using Phage Display Peptide Library

    Institute of Scientific and Technical Information of China (English)

    陈欲晓; 易新元; 曾宪芳; 王林纤; 唐连飞; 章洁; McreynoldsLarry

    2004-01-01

    To select the immunogenic short peptide mimics of male worm origin of Schistosoma japonicum (Sj) and to explore their protection effect against schistosomiasis in mice, the random phage display peptide hbrary of 12 - mer was screened with IgG to soluble male worm antigen of Sj, and the specific positive clones selected through three rounds of screenings were detected by Dot-ELISA, and then injected subcutaneously into mice for vaccination and protection assessment against Sj. It was found that 18 randomly picked phage displayed clones all showed definite antigenicity with various intensities. The pooled phages displayed clones could induce production of specific antibodies and cause 31.72% of worm reduction rate and 51.54 % of egg reduction rate in mice, revealing a significant difference ( P < 0. 001 ) in comparison with those of the controls. It concludes that the short peptide mimics of male worm origin of Sj obtained by affinity screening phage display ptide library can elicit partial protection against this pathogen.

  10. Cloning, expression and protective immunity evaluation of the full-length cDNA encoding succinate dehydrogenase iron-sulfur protein of Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    YU JunLong; WANG ShiPing; LI WenKai; DAI Gan; XU ShaoRui; HE Zhuo; PENG XianChu; ZHOU SongHua; LIU XueQin

    2007-01-01

    1071-bp fragment was obtained from the Schistosoma japonicum (Chinese strain) adult cDNA library after the 3' and 5' ends of the incomplete expression sequence tag (EST) of succinate dehydrogenase iron-sulfur protein of Schistosoma japonicum (SjSDISP) were amplified by the anchored PCR with 2pairs of primers designed according to the EST of SjSDISP and the sequence of multiclone sites of the library vector. Sequence analysis indicated that the fragment was a full-length cDNA with a complete open reading frame (ORF), encoding 278 amino acid residues. The fragment was cloned into prokaryotic expression vector pQE30, and subsequently sequenced and expressed in Escherichia coll.SDS-PAGE and Western-blot analyses showed that the recombinant protein was about 32 kD and could be recognized by the polyclonal antisera from rabbits immunized with Schistosoma japonicum adult worm antigen. Compared with the FCA controls, mice vaccinated with rSjSDISP (test) or rSjGST (positive control) all revealed high levels of specific antibody and significant reduction in worm burden, liver eggs per gram (LEPG), fecal eggs per gram (FEPG) and intrauterine eggs. These results suggest that SjSDISP may be a novel and partially protective vaccine candidate against schistosomiasis. In contrast to the worm burden reduction rate, the higher degree of egg reduction rate in the test group also suggested that SjSDISP vaccine may primarily play a role in anti-embryonation or anti-fecundity immunity.

  11. Cloning, expression and protective immunity evalua- tion of the full-length cDNA encoding succinate dehydrogenase iron-sulfur protein of Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    1071-bp fragment was obtained from the Schistosoma japonicum (Chinese strain) adult cDNA library after the 3′ and 5′ ends of the incomplete expression sequence tag (EST) of succinate dehydrogenase iron-sulfur protein of Schistosoma japonicum (SjSDISP) were amplified by the anchored PCR with 2 pairs of primers designed according to the EST of SjSDISP and the sequence of multiclone sites of the library vector. Sequence analysis indicated that the fragment was a full-length cDNA with a complete open reading frame (ORF), encoding 278 amino acid residues. The fragment was cloned into prokary- otic expression vector pQE30, and subsequently sequenced and expressed in Escherichia coli. SDS-PAGE and Western-blot analyses showed that the recombinant protein was about 32 kD and could be recognized by the polyclonal antisera from rabbits immunized with Schistosoma japonicum adult worm antigen. Compared with the FCA controls, mice vaccinated with rSjSDISP (test) or rSjGST (posi- tive control) all revealed high levels of specific antibody and significant reduction in worm burden, liver eggs per gram (LEPG), fecal eggs per gram (FEPG) and intrauterine eggs. These results suggest that SjSDISP may be a novel and partially protective vaccine candidate against schistosomiasis. In contrast to the worm burden reduction rate, the higher degree of egg reduction rate in the test group also sug- gested that SjSDISP vaccine may primarily play a role in anti-embryonation or anti-fecundity immunity.

  12. Performance of a dipstick dye immunoassay for rapid screening of Schistosoma japonicum infection in areas of low endemicity

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    Peeling Rosanna W

    2011-05-01

    Full Text Available Abstract Background The dipstick dye immunoassay (DDIA, recently commercially available in the People's Republic of China (P.R. China, is a rapid and simple test to detect human antibodies against Schistosoma Japonicum. Its performance and utility for screening schistosome infection in low endemic areas is little known. We therefore carried out a cross-sectional survey in seven villages with low endemicity of schistosomiasis in P.R. China and assessed the performance and utility of DDIA for diagnosis of schistosomiasis. Stool samples were collected and examined by the Kato-Katz method and the miracidium hatching technique. Serum samples, separated from whole blood of participants, were tested by DDIA. Results 6285 individuals aged 6-65 years old participated in this study, with a prevalence of schistosomiasis of 4.20%. Using stool examination as a gold reference standard, DDIA performed with a high overall sensitivity of 91.29% (95% CI: 87.89-94.69% and also a high negative predictive value, with a mean value of 99.29% (95% CI: 98.99-99.58%. The specificity of DDIA was only moderate (53.08%, 95% CI: 51.82-54.34%. Multivariate analysis indicated that age, occupation and history of schistosome infection were significantly associated with the false positive results of DDIA. Conclusions DDIA is a sensitive, rapid, simple and portable diagnostic assay and can be used as a primary approach for screening schistosome infection in areas of low endemicity. However, more sensitive and specific confirmatory assays need to be developed and combined with DDIA for targeting chemotherapy accurately.

  13. Confocal laser scanning microscopic observation on adult Schistosoma japonicum harbored in mice following treatment with single-dose mefloquine.

    Science.gov (United States)

    Xiao, Shu-Hua; Sun, Jun; Xue, Jian

    2012-06-01

    The aim of the present study is to assess the mefloquine-induced alteration of adult Schistosoma japonicum using confocal laser scanning microscopy (CLSM). Eight out of ten mice infected with 60-80 S. japonicum cercariae for 35 days were treated orally with mefloquine at a single dose of 400 mg/kg. Four groups of two mice were killed at 24 h and 3, 7, and 14 days post-treatment, and schistosomes were collected by perfusion from the liver and mesenteric veins, fixed in 70% alcohol, stained with acid carmine, and examined by CLSM. Worms obtained from untreated mice served as controls. Twenty-four hours post-treatment, focal tegument of adult male and female worms, which composed of fine and short villus-like materials, became thicker and longer, or disorder arrangement, while the musculatures beneath the tegument revealed in focal and irregular swelling with various degrees. In the gut of male and female schistosomes, severe dilatation accompanied by swelling, collapse, and peeling of gut mucosa was universal. In the reproductive organs, no apparent alteration in the testis structure of male worms was seen, while in female worms, slight damage to the ovary included loose arrangement of mature ovary cells accompanied by some of them degenerated and collapsed. As to vitelline glands, severe damage, such as swelling, indistinction, fusion or collapse of vitelline cells, and apparent swelling of parenchymal tissues in vitelline gland lobules, was seen. Meanwhile, abnormal ova emerged in the uterus at this time point. Three to 7 days post-treatment, the damage to the worms aggravated either in extent or in severity along with time. In some focally swollen worm body, the parenchymal tissues revealed in severe swelling. In addition, a large piece of degenerated and necrotic parenchymal tissues emerged closed to the severe destructed oral or ventral sucker. In the gut of male and female worms, the major alterations manifested by focal collapse or peeling of mucosa, and

  14. Further studies on mefloquine and praziquantel alone or interaction of both drugs against Schistosoma japonicum in vitro.

    Science.gov (United States)

    Xiao, Shu-hua; Xue, Jian; Zhang, Hao-bing

    2012-03-01

    The aim of the present study is to further understand and analyze the interaction of mefloquine with praziquantel against adult Schistosoma japonicum in vitro. Mice infected with S. japonicum cercariae for 35-37 days were sacrificed, and adult schistosomes were collected by perfusion. Schistosomes were placed to each of 12 wells of a Falcon plate and maintained in RPMI 1640 supplemented by 10% calf serum. For determination of 50% and 95% lethal concentration (LC50 and LC95) of the two drugs in vitro, schistosomes were exposed to mefloquine at concentrations of 1, 2, 3, 4, 5, 6, 7, and 10 μg/mL or praziquantel at concentrations of 0.001, 0.01, 0.05, 0.1, 0.2, 0.5, 1, 10, and 30 μg/mL. The plate was incubated at 37°C in 95% air + 5% CO₂ for 72 h. According to the half-life of oral mefloquine and praziquantel in mice, mefloquine combined with praziquantel simultaneously, mefloquine administered within 1 h after praziquantel and praziquantel administered within 17 h after mefloquine were used to evaluate the effect of mefloquine in combination with praziquantel against S. japonicum in vitro. The results showed that the LC50 and LC95 of mefloquine calculated by the Bliss method were 6.17 μg/mL (95% confidence limits, 5.84-6.517 μg/mL) and 8.703 μg/mL (95% confidence limits, 7.632-9.797 μg/mL), respectively. As to praziquantel, no worm death was seen when schistosomes were exposed to praziquantel at concentrations of 0.005-0.2 μg/mL for 72 h. While in the worms exposed to praziquantel 1, 10, and 30 μg/mL, strong spasmodic contractions of the worm body and vesiculation along the worm surface were observed, but 48-75% of the schistosomes survived the exposure in 72-h incubation. Meanwhile, the number of dead worms that emerged in each group was not proportion to the increasing concentrations. Therefore, it is not appropriate to calculate the LC50 and LC95 of praziquantel. For evaluation of the interaction with the two drugs, praziquantel 0.1 or 0.2

  15. Isolation and Characterization of 44.6 kDa Protein from Schistosoma japonicum Male Worm

    Institute of Scientific and Technical Information of China (English)

    陈欲晓; 易新元; 曾宪芳; 袁仕善; 张顺科; LarryMCREYNOLDS

    2004-01-01

    Soluble male worm antigen of Schistosoma japonicurn (Sj) was investigated for development of new vaccine candidate. SDS--PAGE and Western blotting were performed to compare the difference between soluble antigens from worms of different sex. Mice vaccination with the testing purified protein was followed by Sj cereariae challenge to detect the protective effect against Sj. Sixteen bands were seen for the soluble male worm antigen and 12 for the female worm. In addition, a distinct band of 44.6 kDa from the male worm antigen was observed, and its antigenicity was demonstrated by Western blotting. This 44.6 kDa protein could induce significant worm and egg reduction rate in mice (39.31%, 41.98%, P < 0.001). Inthis study a 44.6 kDa protein was isolated and partially characterized. Its antigenicity, immunogenicity and the partial immune protection suggest its potential vaccine candidte against Sj.

  16. Gene Gun Bombardment with DNA-Coated Golden Particles Enhanced the Protective Effect of a DNA Vaccine Based on Thioredoxin Glutathione Reductase of Schistosoma japonicum

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    Yan Cao

    2013-01-01

    Full Text Available Schistosomiasis, caused by infection with Schistosoma species, remains an important parasitic zoonosis. Thioredoxin glutathione reductase of Schistosoma japonicum (SjTGR plays an important role in the development of the parasite and for its survival. Here we present a recombinant plasmid DNA vaccine, pVAX1/SjTGR, to estimate its protection against S. japonicum in BALB/c mice. The DNA vaccine administrated by particle bombardment induced higher protection than by intramuscular injection. All animals vaccinated with pVAX1/SjTGR developed significant specific anti-SjTGR antibodies than control groups. Moreover, animals immunized by gene gun exhibited a splenocyte proliferative response, with an increase in IFN-γ and IL-4. The recombinant plasmid administrated by gene gun achieved a medium protective efficacy of 27.83–38.83% ( of worm reduction and 40.38–44.51% ( of liver egg count reduction. It suggests that different modes of administering a DNA vaccine can influence the protective efficacy induced by the vaccine. Interestingly, from the enzymatic activity results, we found that worms obtained from pVAX1/SjTGR-vaccinated animals expressed lower enzymatic activity than the control group and the antibodies weakened the enzymatic activity of SjTGR in vitro, too. It implies that the high-level antibodies may contribute to the protective effects.

  17. Deep sequencing-based identification of pathogen-specific microRNAs in the plasma of rabbits infected with Schistosoma japonicum.

    Science.gov (United States)

    Cheng, Guofeng; Luo, Rong; Hu, Chao; Cao, Jie; Jin, Youxin

    2013-12-01

    Circulating microRNAs (miRNAs) have received considerable attention as a novel class of biomarkers for the diagnosis of cancer and as signalling molecules in mediating intercellular communication. Schistosomes, the causative agents of schistosomiasis, live in the blood vessels of a mammalian host in the adult stage. In the present study, we characterized schistosome-specific small RNA populations in the plasma of rabbits infected with Schistosoma japonicum (S. japonicum) using a deep sequencing method and then identified five schistosome-specific miRNAs, including four known miRNAs (Bantam, miR-3479, miR-10 and miR-3096), and one novel miRNA (miR-0001, miRBase ID: sja-miR-8185). Four of the five schistosome-specific miRNAs were also detected by real-time RT-PCR in the plasma of S. japonicum-infected mice. In addition, our study indicated that schistosome Argonaute 2/3 may be an excretory-secretory (ES) protein. In summary, our findings are expected to provide useful information for further development of novel biomarkers for the diagnosis of schistosomiasis and also for deeper understanding of the mechanism of host-parasite interaction.

  18. Comparison of apoptosis between adult worms of Schistosoma japonicum from susceptible (BALB/c mice) and less-susceptible (Wistar rats) hosts.

    Science.gov (United States)

    Wang, Tao; Guo, Xiaoyong; Hong, Yang; Han, Hongxiao; Cao, Xiaodan; Han, Yanhui; Zhang, Min; Wu, Miaoli; Fu, Zhiqiang; Lu, Ke; Li, Hao; Zhao, Zhixin; Lin, Jiaojiao

    2016-10-30

    Schistosomiasis remains a serious public health concern in China. BALB/c mice are susceptible to Schistosoma japonicum infection, whereas the Wistar rats are less susceptible. Apoptosis phenomenon was observed in 42d adult worms of S. japonicum from both rats and mice at the morphologic, DNA, cellular, and gene levels by transmission electron microscopy (TEM), fluorometric terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) analysis, fluorescein isothiocyanate-annexin-V/propidium iodide staining flow cytometry (FCM) analysis, and real-time PCR. The results showed that the apoptotic state in worms from two different susceptible hosts was diverse. Several classical hallmarks of apoptosis, including cell shrinkage, chromatin condensation and lunate marginalization, splitting of the nucleoli, nuclear shrinkage and apoptotic body formation were observed by TEM. TUNEL analysis showed that there were much more apoptosis spots in adult worms from rats than those from mice. Statistical analysis revealed that the degree of apoptosis and percentage of necrotic cells in adult worms from Wistar rats were significantly greater (Pworms from Wistar rats, as compared to those from BALB/c mice. The results obtained in this study collectively demonstrated that differential development of adult S. japonicum in less-susceptible rats and susceptible mice was significantly associated with apoptosis in the worm, and provided valuable information to guide further investigations of the mechanisms governing apoptosis and host interactions in schistosome infection.

  19. The development and prospect of the vaccine against Schistosoma japonicum%日本血吸虫疫苗的研究进展与展望

    Institute of Scientific and Technical Information of China (English)

    景英霞; 邵筱

    2010-01-01

    The study on vaccine against Schistosoma has been included into the program of the treat-ment and prevention on the major diseases of WHO and our country with a great progress. The history of the study on vaccines against Schistosoma has passed from dead vaccine, attenuated vaccine, sub-unit vaccine,gene engineered vaccine, nucleic acid vaccine to muhivalent affiliate vaccine. Recently, the research progress on immunologic mechanism of Schiswsoma and the genome of Schistosoma has played a very important role in the research and development of vaccine against Schistosoma. This article reviewed the development of vaccine against Schistosoma japonicum in recent 10 years.%血吸虫疫苗的研究已经纳入了WHO和我国主要疾病防治规划,并且取得了很大的进展.血吸虫疫苗的研究历史经历了从死疫苗、致弱活疫苗、亚单仲疫苗、基因工程疫苗、核酸疫苗到多价联合疫苗等的探索过程.近年来开展的血吸虫免疫机制和血吸虫基因组的研究对血吸虫疫苗的研制起到了积极的推动作用.该文主要对日本血吸虫疫苗近十年的研究进展作一综述.

  20. Schistosoma japonicum:construction of phage display antibody library and its application in the immunodiagnosis of infection

    Institute of Scientific and Technical Information of China (English)

    陈代雄; 何蔼; 詹希美; 俞慕华; 雷智刚; 孟锦绣; 李卓雅; 梁瑜; 张瑞琳

    2004-01-01

    Background A monoclonal antibody would be an effective tool for the detection of circulating antigens in the serum of patients with schistosomiasis, but the traditional way of producing monoclonal antibodies is not cost-effective. The objective of this study was to find a new method for the large-scale production of monoclonal antibodies against Schistosoma japonicum (Sj).Methods A phage display antibody library for Sj was constructed. To obtain a single-chain variable fragment antibody (scFv) against Sj, the library was screened with metabolic antigens from adult Sj worms (Sj-MAg) using enzyme-linked immunosorbent assay. The soluble scFvs selected were used to detect Sj antigens in the serum of acute and chronic schistosomiasis patients.Results Six positive clones with good reactivity to Sj-MAg were obtained from the phage display antibody library of about 1.07×106 individual clones. Only two of these six clones bound specifically to Sj-MAg and were chosen for further analysis. Specific soluble anti-Sj-MAg scFvs were produced by inducing the 2 clones with isopropyl-D-thiogalactopyranoside. The characteristics of the scFvs were then determined. The results of Western blot showed that these scFvs could bind to Sj-MAg specifically and had a molecular weight of about 31 kD. When testing serum from schistosomiasis patients with one of the two specific scFvs, its sensitivity was found to be 60% and 37% in acute and chronic patients, respectively, with a specificity of 90%. When the two specific scFvs were combined, their sensitivity was found to be 75% and 57% in acute and chronic patients, respectively, with a specificity of 85%.Conclusions The results indicate that the scFvs are potentially useful for the diagnosis of schistosomiasis. The library construction also provides a useful tool for the further screening of other antibodies for both diagnostic and immunotherapeutic applications and for epitope analysis and vaccine design.

  1. Specific anti-glycan antibodies are sustained during and after parasite clearance in Schistosoma japonicum-infected rhesus macaques

    Science.gov (United States)

    Yang, Y. Y. Michelle; Li, Xiao Hong; Brzezicka, Katarzyna; Reichardt, Niels-Christian; Wilson, R. Alan; van Diepen, Angela

    2017-01-01

    Background Human immunity to Schistosoma infection requires many years of exposure, and multiple infections and treatments to develop. Unlike humans, rhesus macaques clear an established schistosome infection naturally at the same time acquiring immunity towards re-infection. In macaques, schistosome egg production decreases after 8 weeks post-infection and by week 22, physiological impairment of the worm caused by unclarified antibody-mediated processes is observed. Since strong antibody responses have been observed against schistosome glycan antigens in human and animal infections, we here investigate if anti-glycan antibodies are associated with immunity against schistosome infections in macaques. Methods We used a microarray containing a large repertoire of glycoprotein- and glycolipid-derived glycans from different schistosome life stages to analyse anti-glycan serum IgG and IgM from S. japonicum-infected macaques during the course of infection and self-cure. We also used an in vitro schistosomula assay to investigate whether macaque sera containing anti-glycan antibodies can kill schistosomula. Conclusions/significance Antibody responses towards schistosome glycans at week 4 post-infection were dominated by IgM while IgG was high at week 8. The profound increase in IgG was observed mainly for antibodies towards a large subset of glycans that contain (multi-)fucosylated terminal GalNAcβ1-4GlcNAc (LDN), and Galβ1-4(Fucα1–3)GlcNAc (LeX) motifs. In general, glycans with a higher degree of fucosylation gave rise to stronger antibody responses than non-fucosylated glycans. Interestingly, even though many IgG and IgM responses had declined by week 22 post-infection, IgG towards O-glycans with highly fucosylated LDN motifs remained. When incubating macaque serum with schistosomula in vitro, schistosomula death was positively correlated with the duration of infection of macaques; macaque serum taken 22 weeks post-infection caused most schistosomula to die

  2. 日本血吸虫DNA疫苗联合免疫的研究进展%Research Progress on combined immunization of DNA vaccine against Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    冯其梅; 张树菊; 汪世平

    2012-01-01

    疫苗作为预防和控制传染病的重要手段,已成功的控制了很多传染病的流行.因此,许多研究者希望研制一种安全有效的血吸虫病疫苗,以期达到控制血吸虫病流行的目的.近年来,鉴于单价疫苗有限的免疫效果,人们开始进行日本血吸虫DNA疫苗联合免疫的研究.本文对日本血吸虫DNA疫苗联合免疫的研究资料进行了归纳整理,发现日本血吸虫DNA疫苗联合免疫涉及鸡尾酒式DNA混合疫苗、双价和多价DNA联合疫苗以及佐剂研究等方面,比较减虫卒、减卵率等指标,联合免疫疫苗的效果明显优于单价DNA疫苗的效果,而且具有潜在的实际应用价值.%As an important mean of prevention and control of infectious diseases, vaccine has been successfully took control. Therefore, in order to control the prevalence of sehistosomiasis japonica, many scientists want to develop a safe and effective vaccine against Schistosoma japonicum. Recently, in view of the limited immune effect of monovalent vaccine, people have started to study combined immunization of DNA vaccine of Schistosoma japonicum. This paper review the development of combined immunization of DNA vaccine against Schistosoma japonicum , and found that combined DNA vaccines include cocktail hybrid DNA vaccines, bivalent and multivalent DNA vaccines, and DNA vaccines with adjuvant mainly. They induced higher worm reduction rate and liver egg reduction rate, and would be well worthy of to be study.

  3. Vaccination of Goats with 31 kDa and 32 kDa Schistosoma japonicum Antigens by DNA Priming and Protein Boosting

    Institute of Scientific and Technical Information of China (English)

    Lianfei Tang; Zhijun Zhou; Yuxiao Chen; Yonghui Luo; Linqian Wang; Liyu Chen; Fushen Huang; Xianfang Zeng; Xinyuan Yi

    2007-01-01

    Two Schistosoma japonicum vaccine candidate antigens Sj 31 and Sj 32, which have shown particular promise to induce protective immunity in mice, were used to immunize goats by using a DNA priming-protein boosting strategy in present work. DNA vaccine formulations of the two antigens (VRSj31 and VRSj32) were produced and injected intramuscularly twice at a 2-week interval and then recombinant proteins (rSj31 and rSj32) together with Freund Complete Adjuvant (FCA) were used to boost the goats. The experiment was repeated in different batche cercariae. A strong anamnestic antibody response was induced after boost. A significant reduction of liver egg counts and miracidial hatching was showed in both experiments. Significant protections against challenge infection were elicited with 31.6% of percentage reduction for worm recovery in the second experiment and 20.9% in the first experiment, respectively.

  4. Identification of Immunodominant Th1-type T cell Epitopes from Schistosoma japonicum 28 kDa Glutathione-S-transferase, a Vaccine Candidate

    Institute of Scientific and Technical Information of China (English)

    Guang-Fu LI; Guan-Ling WU; Yong WANG; Zhao-Song ZHANG; Xin-Jun WANG; Min-Jun JI; Xiang ZHU; Feng LIU; Xiao-Ping CAI; Hai-Wei WU

    2005-01-01

    Th1-type cytokines produced by the stimulation of Th1-type epitopes derived from defined schistosome-associated antigens are correlated with the development of resistance to the parasite infection.Schistosoma mansoni 28 kDa glutathione-S-transferase (Sm28GST), a major detoxification enzyme, has been recognized as a vaccine candidate and a phase Ⅱ clinical trial has been carried out. Sheep immunized with recombinant Schistosoma japonicum 28GST (Sj28GST) have shown immune protection against the parasite infection. In the present study, six candidate peptides (P1, P2, P3, P4, P7 and P8) from Sj28GST were predicted, using software, to be T cell epitopes, and peptides P5 and P6 were designed by extending five amino acids at the N-terminal and C-terminal of P1, respectively. The peptide 190-211 aa in Sj28GST corresponding to the Th1-type epitope (190-211 aa) identified from Sm28GST was selected and named P9.The nine candidate peptides were synthesized or produced as the fusion protein with thioredoxin in the pET32c(+)/BL21(DE3) system. Their capacity to induce a Th1-type response in vitro was measured using lymphocyte proliferation, cytokine detection experiments and flow cytometry. The results showed that P6(73-86 aa) generated the strongest stimulation effect on T cells among the nine candidate peptides, and drove the highest level of IFN-γ and IL-2. Therefore, P6 is a functional Th1-type T cell epitope that is different from that in Sm28GST, and will be useful for the development of effective vaccines which can trigger acquired immunity against S. japonicum. Moreover, our strategy of identifying the Th1-type epitope by a combination of software prediction and experimental confirmation provides a convenient and cost-saving alternative approach to previous methods.

  5. Evaluation of banked urine samples for the detection of circulating anodic and cathodic antigens in Schistosoma mekongi and S. japonicum infections: a proof-of-concept study.

    Science.gov (United States)

    van Dam, Govert J; Odermatt, Peter; Acosta, Luz; Bergquist, Robert; de Dood, Claudia J; Kornelis, Dieuwke; Muth, Sinuon; Utzinger, Jürg; Corstjens, Paul L A M

    2015-01-01

    In Asia, Schistosoma japonicum is the predominant schistosome species, while Schistosoma mekongi is confined to limited foci in Cambodia and Lao People's Democratic Republic. While the People's Republic of China has been successful in controlling schistosomiasis, the disease remains a major public health issue in other areas. In order to prioritise intervention areas, not only accurate diagnosis is important but also other factors, such as practicality, time-efficiency and cost-effectiveness, since they strongly influence the success of control programmes. To evaluate the highly specific urine-based assays for the schistosome circulating cathodic antigen (CCA) and the circulating anodic antigen (CAA), banked urine samples from Cambodia (n=106) and the Philippines (n=43) were examined by the upconverted phosphor lateral flow (UCP-LF) CAA assay and the point-of-care (POC)-CCA urine assay. Based on 250 μl urine samples, UCP-LF CAA sensitivity outcomes surpassed a single stool examination by the Kato-Katz technique. The banked urine samples in the current study did not allow the evaluation of larger volumes, which conceivably should deliver considerably higher readings. The sensitivity of a single urine POC-CCA was in the same order as that of a single Kato-Katz thick smear examination, while the sensitivity approached that of triplicate Kato-Katz when a combination of both CAA and CCA assays was used. The promising results from the current proof-of-concept study call for larger investigations that will determine the accuracy of the urine-based CCA and CAA assays for S. mekongi and S. japonicum diagnosis.

  6. Gene Transcription Profile in Mice Vaccinated with Ultraviolet-attenuated Cercariae of Schistosoma japonicum Reveals Molecules Contributing to Elevated IFN-γLevels

    Institute of Scientific and Technical Information of China (English)

    Xiang ZHU; Feng LIU; Chuan SU; Guan-Ling WU; Zhao-Song ZHANG; Min-Jun JI; Hai-Wei WU; Yong WANG; Xiao-Ping CAI; Lei ZHANG; Shu-Ying HU; Lin-Lin FU

    2005-01-01

    Vaccination with ultraviolet-attenuated cercariae of Schistosoma japonicum induced protective immunity against challenge infection in experimental animal models. Our preliminary study on the transcription levels of IFN-γ and IL-4 in splenic CD4+ T cells revealed that attenuated cercariae elicited predominantly a Thl response in mice at the early stage, whereas normal cercariae stimulated primarily Th2dependent responses. Further analysis on the gene profile of the skin-draining lymph nodes demonstrated that the levels of IFN-γ were significantly higher in vaccinated mice than those in infected mice at day 4, 7 and 14 post-vaccination or post-infection. However, for IL-12 and IL-4, the potent inducers of Th l and Th2 responses, respectively, as well as IL-10, there were no differences over the course of the experiment between the infected and vaccinated mice. To explore the underlying factors that may potentially contribute to elevated IFN-γ in vaccinated mice, the mRNA profiles of the skin-draining lymph nodes at day 4 postexposure were compared using oligonucleotide microarrays. Within the 847 probe sets with increased signal values, we focused on chemokines, cytokines and relevant receptors, which were validated by semi-quantitative RT-PCR. A comprehensive understanding of the immune mechanisms of attenuated cercariae-induced protection may contribute to developing efficient vaccination strategies against S. japonicum, especially during the early stage of infection.

  7. Oral delivery of the Sj23LHD-GST antigen by Salmonella typhimurium type III secretion system protects against Schistosoma japonicum infection in mice.

    Directory of Open Access Journals (Sweden)

    Guo Chen

    2011-09-01

    Full Text Available BACKGROUND: Schistosomiasis japonica is a zoonotic parasitic disease and oral vaccine delivery system would be benefit for prevention of this disease. Although attenuated salmonella has been used as an antigen expression vector for oral vaccine development, the membrane-bound vacuoles in which bacteria reside hinders the presentation of expressed heterologous antigens to the major histocompatibility complex (MHC molecules. The present work used an attenuated Salmonella typhimurium strain VNP20009 to secretory expression of Sj23LHDGST bivalent antigen from Schistosoma japonicum and tested the protective efficacy against S. japonicum infection in orally immunized mice. METHODOLOGY/PRINCIPAL FINDINGS: Promoters (nirB or pagC were used to express the antigen (Sj23LHDGST and the Salmonella type III or α-hemolysin secretion system was employed to secrete it. The immunoblotting analysis and fluorescent microscopy revealed that the antigen was effectively expressed and delivered to the cytosol of macrophages in vitro. Among recombinant vaccine strains, an engineered VNP20009 which expressed the antigen by nirB promoter and secreted it through type III secretion system (nirB-sopE(1-104-Sj23LHD-GST efficiently protected against S. japonicum infection in a mouse model. This strain elicited a predominantly IgG(2a antibody response and a markedly increase in the production of IL-12 and IFN-γ. The flow cytometric analysis demonstrated that this strain caused T cell activation as evidenced by significantly increased expression of CD44 and CD69. CONCLUSION/SIGNIFICANCE: Oral delivery of antigen by nirB-driven Salmonella typhimurium type III secretion system is a novel, safe, inexpensive, efficient and convenient approach for schistosome vaccine development.

  8. Schistosoma japonicum risk in Jiangsu province, People’s Republic of China: identification of a spatio-temporal risk pattern along the Yangtze River

    Directory of Open Access Journals (Sweden)

    Kun Yang

    2013-11-01

    Full Text Available The risk for Schistosoma japonicum infection in Jiangsu province, People’s Republic of China, was investigated by a mouse bioassay. Various investigations were conducted in the period 2009-2011 with the presentation here representing the summary of the results from 45-50 sites in the marshlands along the Yangtze River’s course through the province. Indices representing three aspects of the infection were collected to assess risk: (i the proportion of sentinel points where at least one mouse infection was recorded; (ii the proportion of infected mice at each of these sites; and (iii the average worm burdens. Directional distribution analysis and scan statistics were used to explore the spatio-temporal risk pattern. The spatial distribution was oriented along the Yangtze River and the directional distributions for the proportion of infected mice and mean worm burdens were similar for the positive sentinel sites. Four statistically significant clusters were detected in 2009, but only one in 2010 and 2011, respectively. Temporal windows for infection risk were seen in June and September. The study illustrates the utility of spatio-temporal analysis in assessing the risk for schistosomiasis. This approach should be useful with respect to surveillance and response that can be expected to be increasingly applied when moving from morbidity control to transmission control.

  9. 日本血吸虫和斯氏并殖吸虫的rDNA-ITS2遗传多态性研究%Genetic polymorphism of rDNA-ITS2 of Schistosoma japonicum and Paragonimus skrjabini

    Institute of Scientific and Technical Information of China (English)

    何谐; 张锡林; 牛靖萱; 陈琳

    2009-01-01

    Objective To analyze the genetic variation of rDNA-ITS2 of Schistosoma japonicum and Para-gonimus skrjabini in order to avoid the cross reactions in immune diagnosis between schistosomiasis and paragon-imiasis. Methods The 2 species of fluke were collected, and then the genomic DNA was extracted, to amplify the gene of rDNA-ITS2 by polymerase chain reaction (PCR) and then sequenced. Multiple sequences were aligned by Clusterx, the genetic distance was analyzed by MEGA4, and the phylogentic tree of rDNA-ITS2 genes of Schistosoma and Paragonimus constructed with MP and NJ method. The motifis of the rDNA-ITS2 gene sequences of Schistosoma and Paragonimus were analyzed with DNAsis. Results The results of phylogentic trees of rDNA-ITS2 genes of Schistosoma and Paragonimus were similar. The genetic distance between Schisto-soma japonicum and Paragonimus skrjabini was longer, though there was 42. 7% similarity between their rDNA-ITS2 gene. There were 3 same transcription factors in the sequences of Schistosoma japonicum as well as Para-gonimus skrjabini, AP_2_CS6, bHLH_CS and CAP_site. Conclusion Though there are obviously differences between the Schistosoma japonicum and Paragonimus skrjabini at the level of gene, they have significantly simi-larity in transcription factors of rDNA-ITS2 sequences. It may provide valuable theoretical information for studies on cross-reacting antigen between Schistosoma japonicum and Paragonimus skrjabini at gene level.%目的 分析日本血吸虫和斯氏并殖吸虫的核糖体DNA第二内转录间隔区(rDNA-ITS2)基因的遗传变异.方法 采集2种吸虫成虫标本,提取其基凶组DNA,PCR特异性扩增rDNA-ITS2基因并测序;Clusterx软件进行多序列对比;MEGA 4软件计算遗传距离,NJ法和MP法构建系统进化树;DNAsis软件对rDNA-ITS2序列片段进行模序分析.结果 2种方法构建的遗传系统进化树基本结构相似,日本血吸虫与斯氏并殖吸虫间存在较远的遗传距离,但二吸虫ITS2

  10. MORPHOMETRICS OF TWO DIMENSIONS ON SCHISTOSOMA JAPONICUM EGG%日本血吸虫虫卵的二维数据测量

    Institute of Scientific and Technical Information of China (English)

    彭善友; 黎辉; 等

    2001-01-01

    目的 设计人门静脉内日本血吸虫虫卵过滤器孔眼的大小。 方法 采用图像分析仪测量虫卵的周长、面积、长径、短径、等效直径、圆球度。依据长径、短径数据算出虫卵的体积和均值。 结果 长径最大值84.55 μm,最小值39.19 μm;短径最大值57.22 μm,最小值23.2 μm。体积最大值131 039 μm3,最小值14 423 μm3。 结论 长径99%范围内的最小值45.49 μm,短径99%范围内的最小值27.43 μm,体积99%范围内的最小值17 817.59 μm3。本研究结果为设计人门静脉内日本血吸虫虫卵过滤器孔眼的大小提供了科学依据。%Objective To design the hole size of a filter for Schistosoma japonicum egg in human portal vein, measuring relative data of the eggs is needed. Methods The eggs were gotten form the rabbits infected with schistosomiasis japonicum. The perimeter, area, long diameter, short diameter, eguivalent diameter and sphere grade of each egg were measured by image analyser, then the volume of eggs and its average were worked out according to the long diameter and short diameter. Results Long diameter maximum is 84.55 μm, its minimum is 39.19 μm, short diameter max is 57.22 μm, its minimum is 23.2 μm, volume maximum is 131 039 μm3 and the minimum is 14 423 μm3. Conclusion Minimum in 99% range of the long diameter is 45.49 μm, minimum is 99% range of the short diameter is 27.43 μm, minimum in 99% range of the volume is 17 817.59 μm3. The results supplied scientific warranty for designing the hole size of the filter.

  11. 日本血吸虫非编码小RNA研究进展%Advances in small noncoding RNAs of schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    阳青兰(综述); 肖建华(审校)

    2016-01-01

    Small noncoding RNAs,a growing family of regulatory RNAs that includes micro-RNAs,endogenous small inter-fering RNA,Piwi-interacting RNA and competitive endogenous RNA,have recently emerged as cruicial components in many biological processes. Recent findings have provided the small noncoding RNAs expression to undergo dramatic changes in the schistosomiasis. Accumulated evidence suggests that host miRNAs may be involved in regulating the pathogenesis of schistosomiasis (miR-454 and miR-203). MiR-223 and miR-451 may be involved in regulating the development of schisto-some. Besides,recent work implies that schistosome-derived miRNAs may play critical regulatory roles in schistosome de-velopment and sexual maturation,egg production,as well as the pathogenesis of schistosomiasis (sja-let-7,sja-bantam). Moreover,sja-miR-3479 and sja-miR-0001 could be used as biomarkers for schistosomiasis diagnosis. Here,we summarize recent progress in regarding small noncoding RNAs in schistosoma japonicum. This may be helpful in search for potential new drugs,and for biomarkers in control of schistosoma japonicum infection leading to schistosomiasis in the future.%非编码小RNA是一类通过调节基因的表达从而影响生命活动的小分子,主要包括微小RNA、内源性小干扰RNA、Piwi蛋白相互作用的RNA及竞争性内源RNA。研究发现日本血吸虫感染动物后,宿主和虫体的非编码小RNA的表达水平均发生了改变。宿主源的非编码小RNA主要为微小RNA,其中miR-454和miR-203与日本血吸虫病发生发展有关,miR-223和miR-451与日本血吸虫自身生长发育有关;日本血吸虫非编码小 RNA包括微小RNA、内源性小干扰RNA,这些非编码小RNA在日本血吸虫的生长、发育、性成熟及产卵中发挥着重要作用,如sja-let-7、sja-bantam,而且像sja-miR-3479和sja-miR-0001还可以诊断日本血吸虫病。就日本血吸虫非编码小RNA的研究进展进行综述,为日本血吸虫病的防治提供参考。

  12. IL-6RFP can protect liver function in the model of mice infected with Schistosoma japonicum%IL-6RFP 在小鼠血吸虫感染中对肝细胞的保护作用

    Institute of Scientific and Technical Information of China (English)

    平春光; 程海燕; 高闻达; 计永胜; 刘淼; 任翠平; 邵延靖; 华梦晴; 沈际佳

    2015-01-01

    necrosis factor-α (TNF-α) and chemokine (C-X-C motif) ligand 1 (CXCL1)mRNA levels in mouse liver tissue were detected by RT-PCR.Alanine aminotransferase (ALT)and as-partate transaminase (AST)were measured by the continuous monitoring method.Results HepG2 cells experi-ments showed that mIL-6RFP could reduce the effect of IL-6 on HepG2 cells,fibrinogen (FGG)and haptoglobin (HP)was significantly reduced (P <0.05).In the BALB /c mice infected with Schistosoma japonicum,when IL-6 was blocked the levels of ALT and AST were decreased significantly (P <0.05)compared with that of the solvent group,while there was no significant difference on granuloma formation.Conclusion In the BALB /c mice infec-ted with Schistosoma japonicum,mIL-6RFP can significantly improve the liver function but no obvious effect on granuloma formation.

  13. Effect of mefloquine administered orally at single, multiple, or combined with artemether, artesunate, or praziquantel in treatment of mice infected with Schistosoma japonicum.

    Science.gov (United States)

    Xiao, Shu-hua; Mei, Jing-yan; Jiao, Pei-ying

    2011-02-01

    The purpose of the study is to explore the efficacy of mefloquine administered orally at single, multiple doses, or in combination with artesuante, artemether, or praziquantel in mouse--Schistosoma japonicum model. A total of 205 mice were divided into 4 batches and each batch of mice was infected percutaneously with 40 S. japonicum cercariae for 35 days. The infected mice were treated orally with mefloquine at single doses, multiple daily doses, or combined with artesunate, artemether, or praziquantel, while infected but untreated mice served as control. All treated animals were killed 4 weeks post-treatment for assessment of effect. When infected mice were treated orally with mefloquine at single or multiple daily doses under the same total dose levels, the tendency to decrease the efficacy was seen. Particularly, when a lower single dose of 100 mg/kg was divided equally into five daily doses of 20 mg/kg, the efficacy decreased statistically significant (Pmefloquine or artesuante at a single dose of 100 mg/kg, a moderate effect against schistosomes was observed. No further significant reduction of total and female worm burdens was seen, when the two drugs combined together at the same dose level. On the other hand, administration of mefloquine combined with artesunate at single dose of 50 mg/kg, which exhibited no effect against schistosomes, resulted in significant reduction of total and female worm burdens in comparison with the groups treated with mefloquine and artesunate alone at the same dose level. Similar results were observed in treatment of infected mice with mefloquine in combination with artemether at the smaller dose of 50 mg/kg. The total worm burden was significantly lower than that of control and the female worm burden was also significant lower than that of groups treated with mefloquine and artemether alone. Interestingly, in administration of mefloquine 100 mg/kg combined with artemether 100 mg/kg to the infected mice, all female worms were

  14. Comparative Study on the Immunogenicity between Recombinant MS-Sj26GST Vaccine and Recombinant BCG-Sj26GST Vaccine in Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    戴五星; 高红; 黄海浪; 袁野; 胡佳杰; 皇甫永穆

    2003-01-01

    The BALB/c mice were immunized with rMS-Sj26GST and rBCG-Sj26GST vaccine inSchistosoma japonicum by subcutaneous injection. After they were immunized for 8 weeks, the eye-balls were removed to get blood and macrophages of abdominal cavity and spleen cells were harves-ted. The lymphocytic stimulating index (SI) was used to measure the cellular proliferating abilityand NO release was used to measure the phagocytic activity of the macrophages. By using ELISAkit, the levels of interleukin-2 (IL-2) and interferon-γ(IFN-γ) in serum and the splenic lymphocyt-ic cultured supernatant were detected. The results showed that after the mice were immunized with106 CFU of rMS-Sj26GST and rBCG-Sj26GST vaccine separately by subcutaneous injection, prolif-erating ability of splenic lymphocytes in the mice showed no difference (P>0.05), but both weresignificantly increased as compared with that in the control group(P<0.05); The contents of NOin the intraperitoneal macrophages of rMS-Sj26GST vaccine group were significantly lower than inthe control group (P<0. 001) and rBCG-Sj26GST vaccine group (P<0. 01); The levels of serumIL-2 in the rMS-Sj26GST vaccine group were significantly increased as compared with that in thecontrol group (P<0. 001), vector group (P<0.01) and rBCG-Sj26GST vaccine group (P<0.05);The contents of serum IFN-γ in the rMS-Sj26GST vaccine group were significantly increased ascompared with that in the control group (P<0.01) and rBCG-Sj26GST vaccine group (P<0.05).The contents of IFN-γ in the cultured supernatant were significantly lower than those of rBCG-Sj26GST vaccine group (P<0. 001), but were significantly increased as compared with that in thecontrol group (P<0.01). It was indicated that both vaccines could enhance the immune response ofthe mice, but rMS-Sj26GST vaccine had stronger immunogenicity than rBCG-Sj26GST vaccine.

  15. 水牛和猪感染日本血吸虫后的自愈现象及其机制%The self-cure phenomenon in buffalo and pig infected with Schistosoma japonicum and its mechanism

    Institute of Scientific and Technical Information of China (English)

    吕美云; 李宜锋; 林丹丹

    2010-01-01

    动物感染血吸虫后的自愈现象是指动物在感染血吸虫一段时间后虫负荷数急剧下降、虫体自然清除的现象.水牛和猪作为主要的血吸虫病传染源,在血吸虫病流行传播中起到重要作用.已有研究表明,水牛和猪感染日本血吸虫后有自愈现象,阐明其发生机制对于防治血吸虫病和研制血吸虫疫苗均有重要意义.该文主要围绕水牛、猪感染日本血吸虫后产生的与自愈有关的实验和现场研究作一综述,并对其发生的机制进行探讨.%Self-cure refers to the phenomenon that the number of worms declines sharply and parasites are cleared up naturally after animals infected with Schistosoma japonicum for some time. Buffaloes and pigs play an important role in schistosomiasis transmission as the main sources of infection. Some studies have shown that self-cure occurs in water buffaloes and pigs infected with S. japonicum. It has great significance to clarify the mechanisms behind the phenomenon of self-cure for prevention and control of schistosomiasis and development of anti-schistosomiasis vaccine. This paper summarized the experimental research and field epidemiologi-cal studies around self-cure in water buffaloes and pigs infected with 5. japonicum and discussed the mechanisms of self-cure.

  16. Significance of higher drug concentration in erythrocytes of mice infected with Schistosoma japonicum and treated orally with mefloquine at single doses.

    Science.gov (United States)

    Tao, Yi; Xue, Jian; Jiang, Bin; Zhang, Hao-Bing; Xiao, Shu-Hua

    2015-12-01

    The purpose of the present study is to understand the pharmacokinetic feature of mefloquine measured by erythrocytes and plasma in Schistosoma japonicum (S. j.)-infected mice and non-infected mice after oral administration of the drug at single doses. A high-performance liquid chromatography (HPLC) method was used to measure the plasma and erythrocyte concentrations of mefloquine at varying intervals posttreatment. Our results demonstrated that in non-infected mice treated orally with mefloquine at an ineffective dose of 50 mg/kg or effective dose of 200 mg/kg for 2-72 h, the erythrocyte-to-plasma ratios of mefloquine were 5.8-11.2 or 2-14.2. On the other hand, in S. j.-infected mice treated with the same single doses of the drug, the erythrocyte and plasma drug concentration ratios were 3.1-4.6 or 2.9-8.5, manifesting that either in infected mice or in non-infected mice that received oral mefloquine resulted in higher concentration of mefloquine in erythrocytes than that in plasma. Unexpectedly, under oral administration of mefloquine at a higher single dose of 200 mg/kg, the pharmacokinetic parameter C max values for plasma from S. j.-infected and non-infected mice were 1.6 ± 0.3 and 2.0 ± 0.4 μg/mL, respectively, which were below the determined in vitro LC50 (50 % lethal concentration) value of 4.93 μg/mL. Therefore, the plasma concentration of mefloquine may display a little effect against schistosomes during the treatment. Although the values of T 1/2 and AUC0-∞ for erythrocytes were significantly longer and higher in infected mice than those of corresponding non-infect mice that received the same single mefloqine dose of 50 mg/kg, the C max value was only 2.6 ± 0.4 μg/mL lower than the determined in vitro LC50, which may explain why this low single dose is ineffective against schistosomes in vivo. After administration of higher mefloquine dose of 200 mg/kg, the C max value for erythrocytes in infected mice was 30 % (7.4 ± 0

  17. Molecular characterization of thyroid hormone receptor beta from Schistosoma japonicum and assessment of its potential as a vaccine candidate antigen against schistosomiasis in BALB/c mice

    Directory of Open Access Journals (Sweden)

    Qiu Chunhui

    2012-08-01

    Full Text Available Abstract Background Thyroid hormones (TH modulate growth, development and differentiation and metabolic processes by interacting with thyroid hormone receptors (THRs. The purpose of this study was to identify a novel thyroid hormone receptor beta encoding gene of Schistosoma japonicum (SjTHRβ and to investigate its potential as a vaccine candidate antigen against schistosomiasis in BALB/c mice. Methods The full-length cDNA sequence of SjTHRβ, its gene organization, and its transcript levels were characterized, and the phylogenetic relationship between THR, RAR and RXR from other organisms were analysis, the ability of this protein binding to a conserved DNA core motif, and its potential as a vaccine candidate antigen against schistosomiasis in BALB/c mice were evaluated. Results The SjTHRβ cDNA was cloned, verified by 5’ and 3’ Rapid Amplification of cDNA Ends and shown to be polyadenylated at the 3’end, suggesting the transcript is full-length. SjTHRβ is homologous to THRs from other species and has a predicted conservative DNA binding domain and ligand binding domain that normally characterizes these receptors. A comparative quantitative PCR analysis showed that SjTHRβ was the highest expressed in 21d worms and the lowest in 7 d and 13 d schistosomula. The cDNA corresponding to DNA binding domain (SjTHRβ-DBD and ligand binding domain (SjTHRβ-LBD were cloned and subsequently expressed in E coli. The expressed proteins were used to immunize mice and generate specific serum against recombinant SjTHRβ (rSjTHRβ. Western blotting revealed that anti-rSjTHRβ-LBD serum recognized two protein bands in extracts from 21 d worm with molecular sizes of approximately 95 kDa and 72 kDa. Electrophoretic mobility shift assay (EMSA analysis showed that rSjTHRβ-DBD could bind to a conserved DNA core motif. Immunization of BALB/c mice with rSjTHRβ-LBD could induce partial protective efficacy(27.52% worm reduction and 29.50% liver eggs

  18. Risk factors of Schistosoma japonicum infection in Xingzi County%星子县居民血吸虫感染危险因素分析

    Institute of Scientific and Technical Information of China (English)

    肖国良; 翟敏玲; 陶波; 姜秋林; 刘建平; 习青华; 周宪民; 卢曲琴

    2016-01-01

    目的:分析江西省星子县居民血吸虫感染的危险因素。方法2013年采用整群抽样的方法从江西省星子县血吸虫病流行区随机抽取6个行政村进行调查,并采用Kato⁃Katz法对这6个村≥5岁的全部常住居民进行病原学检查;应用单因素χ2检验和多因素Logistic回归模型对影响血吸虫病流行的相关危险因素进行分析。结果2013年共粪检调查2050人,查出粪检阳性146人,血吸虫感染率为7.1%。单因素χ2检验显示,不同性别、年龄、文化程度、职业人群血吸虫感染率差异均有统计学意义(χ2=26.485、16.836、25.700、90.805,P均<0.05)。多因素Logistic回归分析显示,男性感染血吸虫的风险是女性的3.041倍;小学及以下文化程度人群血吸虫感染风险是大专及以上文化程度人群的8.870倍;初中和高中文化程度人群感染血吸虫的风险分别是大专及以上文化程度人群的5.598倍和2.995倍;渔民血吸虫感染的可能性最高,是其他职业的3.053倍。结论影响星子县居民血吸虫感染的危险因素主要包括性别、职业和文化程度;应强化血吸虫病健康教育,加强疫水接触防护工作,减少血吸虫感染发生。%Objective To explore the risk factors of Schistosoma japonicum infection in the residents in Xingzi County,Ji⁃angxi Province. Methods Six administrative villages from different areas were randomly selected by the cluster sampling meth⁃od as the study sites in Xingzi Country in 2013,and all the residents aged 5 years or above were investigated epidemiologically, and the schistosome infection was surveyed by Kato⁃Katz technique. The risk factors of schistosomiasis were analyzed by using the Chi⁃square test analysis and multivariate Logistic regression model. Results In 2013,there were 2 050 residents received the stool examination and 146 persons were positive,the schistosome infection rate was 7.1%. The Chi

  19. SEQUENCE ANALYSIS AND PRELIMINARY IDENTIFICATION OF MOLECULES WITH EF-HAND DOMAIN OF SCHISTOSOMA JAPONICUM%日本血吸虫含EF手性结构域分子序列的分析和初步鉴定

    Institute of Scientific and Technical Information of China (English)

    张艳丽; 贾侃; 李莹; 苑纯秀; 杨健美; 林矫矫; 冯新港

    2011-01-01

    为分离鉴定日本血吸虫含EF手性结构域分子及分析其序列结构特征,首先找到含有EF—hand结构域的分子,按照序列一级结构进行分类,之后进行生物信息学分析。然后用PCR方法以虫卵和成虫cDNA文库为模板扩增分类后的分子,构建重组质粒,诱导蛋白表达并纯化,选取14个纯化蛋白用Western blot进行初步免疫原性鉴定。结果269个原始含EF-hand结构域分子按照序列一级结构分为70个;成功表达和纯化了49个含EF手性分子,进化树分析将其分为9类;Western blot显示,14个纯化蛋白均可被日本血吸虫感染小鼠阳性血清识别。本研究结果为下一步采用这类分子进行动物保护性效果评价以及保护性免疫机制的研究奠定了基础。%To isolate and identify the molecules with EF-hand domain of Schistosoma japonicum and analyse the sequence structural characteristics of them, the molecules with EF-hand domain were firstly found and classified according to the primary structure, and then analyzed by bioinformatics. PCR was used to amplify the classified molecules with the S.japonicum adult and egg cDNA library. The recombinant plasmids were constructed and the protein with EF-hand domain were expressed and purified. Western blot was used to identify the preliminary immunogenicity of 14 purified proteins. Two hundred and sixty nine original molecules with EF-hand domain were divided into 70 molecules. Forty nine molecules with EF-hand domain were expressed and divided into 9 categories by phylogenetic tree. Western blot analysis showed that 14 purified proteins were specifically recognized by the positive serum from mice infected with S.japonicum. The study provided the basis for further study of immune protection of the molecules.

  20. 室内繁殖东方田鼠感染日本血吸虫的实验研究%Study of closed colony of Microtus fortis infected with Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    成钢; 曾文虎; 王京仁; 王文彬; 李淑红

    2013-01-01

    目的 比较室内繁殖东方田鼠与昆明鼠感染日本血吸虫后不同阶段脏器病变以及体内日本血吸虫发育与存活状况.方法 采用日本血吸虫尾蚴感染健康东方田鼠和昆明鼠,在感染后12、20、40d剖杀,观察并比较东方田鼠以及昆明鼠肝、肾、肺等脏器组织病变及体内日本血吸虫发育与存活情况.结果 日本血吸虫感染后12、20 d昆明小鼠和感染40d后东方田鼠的内脏器官均未出现明显病变;感染12 d及20d后的东方田鼠肝脏、肾脏及脾脏出现明显白色结节,且以感染12d的组织病变更为明显,部分东方田鼠病变组织仅见于肝脏.病理切片显示,病变肝脏和肾脏组织中存在完整日本血吸虫虫体,与正常组织界限分明;而无病变的肝脏和肾脏组织病理切片未发现虫体.结论 室内繁殖东方田鼠感染日本血吸虫后12d免疫反应剧烈且存在明显的个体差异.%Objective To understand the pathological changes of tissues and organs in Microtusfortis infected with Schistosoma japonicum for further insight into the biological dynamic variation and explore immune mechanisms of M.fortis against S.japonicum infection.Methods The healthy M.fortis and mouse were infected with cercariae of S.japonicum through abdominal skin.On the day 12,20 and 40,the M.fortis and mouse were sacrificed by cervical dislocation and dissected.The liver and kidney slices of M.fortis and mouse of the different groups were prepared and stained with hematoxylin eosin stain (HE) by conventional techniques,respectively.Results We did not find obvious lesions on the internal organs of mouse 12 d and 20 d after the infection and the lesions of M.fortis 40 d after the infection.However,white nodules appeared on the liver,kidney and spleen of the M.fortis 12 d and 20 d after the infection,and there were a lot of S.japonicum worms on pathological sections of inflamed liver and kidney tissues,and the boundary between worms and

  1. 日本血吸虫基因组核糖体移码序列的预测分析%Prediction and analysis of frameshifts in the genome of Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    王海彬; 杨忠; 刘锋; 胡薇

    2012-01-01

    目的 预测日本血吸虫基因组中核糖体移码的基因序列并进行鉴定.方法 挑选稳定并能够可靠预测RNA假结结构的软件,编写批量提交数据的程序并结合本地手段进行假结结构预测,计算序列最小自由能从而挑选稳定序列,进一步使用生物信息学软件Fsfinder分析序列中核糖体移码位点,进行开放性阅读框(open reading frame,0RF)分析,筛选出可能产生核糖体移码的日本血吸虫基因序列.利用日本血吸虫蛋白质组数据库中的肽段质谱数据进行比对,寻找对应的肽段信息.结果 从日本血吸虫的8452条基因编码序列中预测出26条可能含有促使核糖体移码假结结构的序列.经过日本血吸虫蛋白质组数据库中的肽段质谱数据进行比对,发现日本血吸虫输入蛋白(Sjimportin)移码之后产生的肽段.结论 整合已有的RNA假结预测软件以及核糖体移码预测软件,建立了日本血吸虫预测核糖体移码序列数据库,并成功获取日本血吸虫蛋白Sjimportin核糖体移码表达证据.%Objective To predict and identify the frameshift sequences in the genome of Schistosoma japonicum.Methods Perl software was used to predict candidate sequences with pseudoknots of S.japonicum automatically.The stable sequences were searched based on the calculation of the minimum free energy and the FSfinder software was used to predict frameshift sites of the sequences,which were compared with proteome database by BLAST.Results From the 8 452 sequences in the S.japonicum database,26 candidate sequences containing both pseudoknot and corresponding frameshift site were selected and the protein Sjimportin bearing frameshift with the evidence in proteome database of S.japonicum was found.Conclusion Combining the RNA pseudoknots prediction software and frameshift prediction software,the database of S.japonicum frameshift has been built and the evidence of frameshift for Sjimportin has been got.

  2. 日本血吸虫糖基化磷脂酰肌醇锚定蛋白的鉴定%Identification of Glycosylphosphatidylinositol-anchored Protein from Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    曹勤燕; 薛艳凤; 沈利

    2012-01-01

    目的 鉴定日本血吸虫的糖基化磷脂酰肌醇锚定蛋白(GPI-AP). 方法 根据曼氏血吸虫的糖基化磷脂酰肌醇(GPI)锚定蛋白Sm200的编码基因(GenBank登录号为XM 002569560.1),运用生物信息学方法寻找日本血吸虫的同源基因,分析后选取基因蛋白质编码区(CDS)的部分基因序列(SjGPIs,长约933 bp)进行PCR扩增,并克隆入原核表达载体pET-28a(+),重组质粒转化大肠埃希菌(E.coli) BL21 (DE3)感受态细胞,异丙基-β-D-硫代半乳糖苷(IPTG)诱导表达,用镍柱Ni-NTA亲和层析纯化重组肽段SjGPIs.用纯化的重组肽段SjGPIs免疫新西兰大耳兔,以制备的重组肽段抗血清检测日本血吸虫GPI锚定蛋白.用磷脂酰肌醇特异性磷脂酶C(PI-PLC)鉴定检测到的蛋白在日本血吸虫虫体上的锚定方式.检测日本血吸虫感染小鼠的白细胞,确定其是否吞噬GPI锚定蛋白. 结果 日本血吸虫的基因组中存在与曼氏血吸虫GPI锚定蛋白Sm200基因的同源基因序列,经比对拼接后获得3495 bp含完整编码蛋白C末端的基因编码序列.以所选基因序列进行肽段原核表达,获得重组质粒pET-28a(+)-SjGPIs.通过对蛋白C末端序列分析、经蛋白质印迹(Western blotting)分析和PI-PLC酶切验证,发现日本血吸虫被膜存在以GPI形式锚定的蛋白,相对分子质量约为Mr 200000,命名为SjGPI200.感染日本血吸虫小鼠的白细胞中可检测到完整的SjGPI200蛋白. 结论 日本血吸虫存在锚定蛋白SjGPI200,并以GPI形式锚定于虫体被膜上.%Objective To identify glycosylphosphatidylinositol (GPI) anchored protein of Schistosoma japonicum. Methods Based on the gene sequence of Schistosoma mansoni GPI anchored protein Sm200 (GenBank Assess No: XM_002569560.1), bioinformatics analysis was performed to find out its homologous gene sequence in S. japonicum, then a selected partial coding sequence (SjGPIs, about 933 bp) from the homologous gene sequence were amplified, and

  3. Research progress on transmission capacity of reservoir host of Schistosoma japonicum%日本血吸虫保虫宿主传播能量研究进展

    Institute of Scientific and Technical Information of China (English)

    汪奇志; 汪天平; 张世清

    2013-01-01

    本文对牛、羊、猪、犬、猫、野鼠等日本血吸虫保虫宿主传播能量的相关研究进行了综述,并结合当前实施的以传染源控制为主的综合防治策略,提出了在人和牛作为传染源的作用逐渐减弱的情况下,对非牛家畜(羊、犬、猪等)和野鼠等动物宿主的实际传播能量进行重新评价,以及探索血吸虫传播环节中相关传播阈值等研究方向.%This paper reviews the researches related to the reservoir hosts of Schistosoma japonicum, including bovine, sheep, dog, cat and wild mouse. Combined with the implementation of the comprehensive control strategy with an emphasis on infection source control in current, it puts forward several future research directions under the condition that the effects of people and bovine as infection sources weakened gradually, such as revaluation on the actual transmission capacity of non-bovine livestock, for example, sheep, dog, pig and wild mouse, as well as discussion on some transmission threshold values in the infection link of Oncomelania snails.

  4. 日本血吸虫原肌球蛋白cDNA的克隆及其在大肠杆菌中表达%Cloning of cDNA encoding Schistosoma japonicum tropomyosin and its expression in Escherichia coli

    Institute of Scientific and Technical Information of China (English)

    曹建平; 刘述先; 宋光承; 徐馀信

    2002-01-01

    Objective To perform cloning of the gene encod ing Chinese Schistosoma japonicum tropomyosin (SjcTM) and its expression in Escherichia coli.Methods SjcTM cDNA fragment, except for 14 amino acids at the amino terminus, was obtained by reverse transcriptase-polymerase chain reaction (RT-PCR) with total RNA extracted from adult worms of S. Japonicum . The RT-PCR product was cloned into T vector and sequenced. The SjcTM cDNA, derived from the constructed TA clone pGEM-SjcTM, was then subcloned into the expressing vector pBV220. After characterization by agarose gel electrophoresis, endonucleases digestion and PCR, the resultant recombinant plasmid was used for expression under the temperature-dependent condition. Results The RT-PCR product, cloned into a Tvector, was sequenced and shown to be 96.5% identical at the nuclei acid level and 98.1 % identical in deduced amino acid sequence to that of S. Mansoni tropomyosi n. The target DNA fragment was then subcloned into a prokaryotic vector pBV220 . Induced expression in E. Coli DH5α cells resulted in a constant level of recombinant protein production. The results of SDS-PAGE and Western blot rev ealed that the molecular weight of non-fusion recombinant protein (rSjcTM) was approximately 32 kDa and could be recognized specifically by a polyclonal antise rum specific for native S. Japonicum tropomyosin (SjcTM). Conclusion The engineering of the cDNA encodingS. Japonicum tropomyosin and its bacterial expression was successfully made.%目的克隆日本血吸虫原肌球蛋白编码基因,并在大肠杆菌中表达.方法抽提日本血吸虫(大陆株)成虫总RNA,经逆转录-聚合酶链反应(RT-PCR)获得编码日本血吸虫原肌球蛋白的cDNA片段,该片段与全序列比较,缺氨基端14个氨基酸.该PCR产物克隆入T 载体并对插入片段进行序列测定后,亚克隆入表达载体pbV220,经琼脂糖凝胶电泳、限制性酶切反应和PCR鉴定后,选择克隆用于温控表达.结果 RT-PCR产物

  5. 日本血吸虫尾蚴及童虫可溶性抗原蛋白质组研究%The soluble antigenic proteome of Schistosoma japonicum cercariae and schistosmula

    Institute of Scientific and Technical Information of China (English)

    马安; 王越; 汤益; 杨再峰; 施晓华; 朱明东; 刘晓龙; 干小仙

    2011-01-01

    应用免疫蛋白质组研究方法筛选、鉴定日本血吸虫尾蚴、童虫可溶性抗原蛋白质。方法日本血吸虫尾蚴可溶性粗抗原(SCAP)、童虫可溶性粗抗原(SLAP)分别用双向凝胶电泳(2-DE)分离蛋白质,每样本同时制3块胶,1块胶进行银染,2块胶通过电转印后再分别用感染兔和正常兔血清作Western印迹分析,确定特异性阳性反应点,再从相应银染胶图上找到匹配的抗原蛋白质点;用MALDI-TOF/TOF串联质谱鉴定抗原蛋白质。结果SCAP、SLAP与感染兔血清免疫反应分别获得94个和68个阳性点,在相应的银染胶图上分别获得33个和31个匹配蛋白质点;用MALDI-TOF/TOF串联质谱鉴定和NCBI数据库检索,鉴定成功率分别为100.0%(20/20)和68.2%(15/22)。已获鉴定的SCAP抗原中蛋白酶占62.5%(10/16),SLAP抗原中蛋白酶占36.4%(4/11);2个抗原蛋白为SCAP和SLAP共有。结论2-DE能有效地分离日本血吸虫可溶性抗原蛋白质,2-D Western印迹法能较好地筛选特异性抗原;2-D Western印迹法阳性点与2-DE胶图蛋白质点匹配率低,低丰度的抗原蛋白质易被漏检;尾蚴和童虫的抗原蛋白质组差异较大。%Objective To screen and identify specific antigenic proteins of Schistosoma japonicum cercariae and schistosomula using immunproteomics approaches. Methods Soluble antigenic proteins of Schistosoma japonicun cercarie (SCAP) and 15 days lung-stage schistosomulum (SLAP) were separated by two-dimensional electrophoresis (2-DE). For each sample, three gels were run in parallel with one gel for silver stain and the other two gels for Western blot using Schistosoma japonicum infected rabbit sera and normal rabbit sera separately. The specific antigenic protein spots were determined on the membrane of Western blot. The matched antigenic protein spots on the sliver stained gels were subsequently analysed by MALDI-TOF/TOF-MS/MS respectively. Results 94 and 68 positive

  6. 白头翁总皂苷对日本血吸虫虫卵毛蚴尾蚴的作用%Effect of Pulsatilla chinensis (Bunge) Regel saponins on eggs, miracidia and cercariae of Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    陈岩勤; 许琼明; 诸葛洪祥; 梁幼生; 李笑然; 杨世林

    2013-01-01

    目的 观察白头翁总皂苷(Pulsatilla chinensis(Bunge)Regel saponins,PRS)对日本血吸虫虫卵、毛蚴、尾蚴的杀灭效果,为白头翁总皂苷作为抗血吸虫新药提供理论和实验依据.方法 采用尾蚴腹部贴片法感染ICR小鼠,感染后42 d的鼠肝脏经研磨、过筛获得虫卵,毛蚴由虫卵孵化,尾蚴由阳性感染钉螺光照逸出,采用0、1、2、3、4、5、6、7、8 μg/ml的PRS药液分别于不同时间作用虫卵、毛蚴、尾蚴.结果 不同浓度PRS及对照药物吡喹酮(PZQ)对日本血吸虫虫卵作用24 h的孵化结果显示,PRS对虫卵孵化的抑制效果略优于PZQ,尤其在4μg/ml浓度时,日本血吸虫虫卵对PRS的作用更敏感.1、2、3、4、5、6、7、8 μg/ml的PRS药液作用于毛蚴30 min后,毛蚴死亡率分别为13.47%、26.05%、60.99%、90.84%、100%、100%、100%、100%.作用于尾蚴30 min后,尾蚴的死亡率分别为5.32%、18.81%、44.7%、76.87%、98.28%、100%、100%、100%.毛蚴、尾蚴的死亡率对PRS的作用时间和浓度有一定依赖性.结论 体外实验显示PRS对日本血吸虫虫卵、毛蚴、尾蚴均有杀伤作用,有望成为新的抗血吸虫药物.%Objective To investigate the effect of Pulsatilla chinensis (Bunge) Regel saponins (PRS) against the eggs, mira-cidia, cercariae of Schistosoma japonicum in vitro and compare its efficacy with praziquantel. Methods ICR mice were infected with the cercariae of S. japonicum by the patching abdominal method. The livers of the mice were grinded, screened, and then the eggs of S. japonicum were obtained 42 days post-infection. The miracidia were hatched by using the eggs, and the cercariae were obtained by using the infected Oncomelania snails on the light. The eggs, miracidia and cercariae of S. japonicum were incubated with 0,1,2,3,4,5,6,7,8 μg/ml PRS for different time, and praziquantel (PZQ) was used as the control. Results PRS suppressed the hatching rates of eggs for

  7. Characterization of a gene family encoding SEA (sea-urchin sperm protein, enterokinase and agrin-domain proteins with lectin-like and heme-binding properties from Schistosoma japonicum.

    Directory of Open Access Journals (Sweden)

    Evaristus Chibunna Mbanefo

    Full Text Available BACKGROUND: We previously identified a novel gene family dispersed in the genome of Schistosoma japonicum by retrotransposon-mediated gene duplication mechanism. Although many transcripts were identified, no homolog was readily identifiable from sequence information. METHODOLOGY/PRINCIPAL FINDINGS: Here, we utilized structural homology modeling and biochemical methods to identify remote homologs, and characterized the gene products as SEA (sea-urchin sperm protein, enterokinase and agrin-domain containing proteins. A common extracellular domain in this family was structurally similar to SEA-domain. SEA-domain is primarily a structural domain, known to assist or regulate binding to glycans. Recombinant proteins from three members of this gene family specifically interacted with glycosaminoglycans with high affinity, with potential implication in ligand acquisition and immune evasion. Similar approach was used to identify a heme-binding site on the SEA-domain. The heme-binding mode showed heme molecule inserted into a hydrophobic pocket, with heme iron putatively coordinated to two histidine axial ligands. Heme-binding properties were confirmed using biochemical assays and UV-visible absorption spectroscopy, which showed high affinity heme-binding (K D = 1.605×10(-6 M and cognate spectroscopic attributes of hexa-coordinated heme iron. The native proteins were oligomers, antigenic, and are localized on adult worm teguments and gastrodermis; major host-parasite interfaces and site for heme detoxification and acquisition. CONCLUSIONS: The results suggest potential role, at least in the nucleation step of heme crystallization (hemozoin formation, and as receptors for heme uptake. Survival strategies exploited by parasites, including heme homeostasis mechanism in hemoparasites, are paramount for successful parasitism. Thus, assessing prospects for application in disease intervention is warranted.

  8. Construction and Identification of Recombinant Baculovirus with Schistosoma japonicum Sj16 Gene%日本血吸虫Sj16基因重组杆状病毒的构建和鉴定

    Institute of Scientific and Technical Information of China (English)

    胡少敏; 王海; 阮志燕; 余新炳; 吴忠道

    2005-01-01

    目的利用杆状病毒-昆虫细胞系统构建日本血吸虫 (Schistosoma japonicum, Sj)Sj16基因重组杆状 病毒.方法从日本血吸虫尾蚴提取总 RNA,通过 RT- PCR扩增出 Sj16基因全编码区序列,将其克隆到载体 pET30a(+ )中,通过 PCR将 Sj16基因连同 pET30a(+ )多克隆位点下游的 6× His· Tag一起扩增出来,插入 donor载体 pFastBacHTa中,构建重组杆状病毒 donor载体 pFastBacHTa- Sj16- His,转化大肠杆菌 DH10Bac- GFP进行转座,提取重组 Bacmid,用 Lipofectin法转染昆虫细胞 Sf9使其包装成有感染性的重组杆状病毒.结果构建了含日本血吸虫 Sj16基因的重组 Bacmid- GFP- Sj16- His,转染 Sf9细胞后,获得了有感染力的重组杆状病毒. 结论成功构建了日本血吸虫 Sj16基因重组杆状病毒,为下一步重组 Sj16蛋白表达和 Sj16基因功能研究打下了基础.

  9. 日本血吸虫成虫吡喹酮用药前后差异表达序列的筛选%Screening of differential expression of expressed sequence tags (ESTs) in adult Schistosoma japonicum treated with praziquantel

    Institute of Scientific and Technical Information of China (English)

    周水茂; 江元山; 陈智; 熊燕; 田俊华; 周敦金; 徐明星; 孙东光

    2011-01-01

    Objective To establish subtracted cDNA libraries using the suppression subtractive hybridization (SSH)technique based on the livers of New Zealand white rabbits infected with Schistosoma japonicum before and after praziquantel treatment, providing the basis for screening the drug-response molecular target during the therapy.Methods New Zealand white rabbits infected with S.japonicum were assigned to two groups to receive praziquantel medication or blank, respectively.The SSH technique was conducted to build the forward and reverse subtracted cDNA libraries using the PCR-Select cDNA Subtraction kit.The differentially expressed ESTs were screened and positive clones were isolated from the libraries for bioinformatic analysis.Results Thirty-nine positive clones were isolated from two subtracted cDNA libraries, including 22 from the forward library and 17 from the reverse library.The analysis revealed that the ESTs were mainly responsible for coding enzymes and proteins associated with protein synthesis and degradation.Conclusion The subtracted cDNA libraries before and after praziquantel treatment for S.japonicum were successfully constructed, laying a solid foundation for further research into the molecular mechanisms of this medication therapy.%目的 应用抑制性消减杂交(SSH)技术构建吡喹酮治疗日本血吸虫感染新西兰大白兔前后肝脏内差减cDNA文库,为筛选吡喹酮治疗过程中日本血吸虫体内药物反应分子靶标奠定基础.方法 日本血吸虫感染新西兰大白兔,吡喹酮治疗者为实验组,未经吡喹酮治疗者为对照组,应用PCR-Select cDNA Subtraction试剂盒进行SSH分析,分别构建正向和反向消减cDNA文库,对文库进行筛选,挑取阳性克隆测序获得差异表达EST或新EST,并进行生物信息学分析.结果 从2个消减文库中共筛选到39个有效阳性克隆,其中正向文库22个,反向文库17个,分析表明这些EST编码主要是一些酶及与蛋白质合成和降

  10. Preliminary investigation on in vitro-cultivation and manipulation of Schistosoma japonicum sporocysts%日本血吸虫胞蚴的体外培养和操作初探

    Institute of Scientific and Technical Information of China (English)

    江艳; 龚仁敏; 杨艺; 李小红; 刘毅; 朱传刚; 陆珂; 郑浩

    2011-01-01

    目的 探究一种体外培养日本血吸虫胞蚴的新方法,并通过体外注射,感染阴性钉螺,培养阳性钉螺.方法 将日本血吸虫虫卵孵化的毛蚴与作为滋养层的草地夜蛾(Spodoptera frugiperda)卵巢细胞(sf9细胞)共培养,观察毛蚴的生长情况,并拍摄生长照片.用显微注射针将3~5只母胞蚴注射到钉螺体内,计算钉螺的存活率及尾蚴的阳性率,用钉螺逸出的尾蚴人工感染昆明鼠,检测尾蚴的活力,用组织化学染色法和PCR方法鉴定尾蚴的相关基因特性.结果 毛蚴培养至第12小时观察到纤毛板已脱落,培养2~3 d的虫体呈不断的伸缩运动,此后母胞蚴维持该状态并不转化成子胞蚴.3次实验的钉螺存活率依次为24.72%、28.23%和57.89%,活螺阳性率依次为4.55%、8.57%和14.29%.阳性钉螺释放的尾蚴具有感染性,且具有与日本血吸虫相同的基因特性.结论 体外培养毛蚴通过显微注射可以人工感染钉螺并逸出有感染活力的尾蚴,为日本血吸虫幼虫的体外操作提供一种新的方法.%Objective To explore a new method of in vitro cultivation of Schistosoma japonicum sporocysts,infection of negative snails through in vitro-injection,and cultivation of positive snails.Methods Isolated miracidia were directly cultivated in sf9 cells-conditioned medium,the growth of miracidia was observed and the growth picture was taken.3-5 mother sporocysts were inoculated into the snails body using a microscopic needle.The survival rate of the snails and the positive rate of cercariae were calculated.Kunming mice were infected with the cercariac and the activity of cercariae Was detected.The related gene characteristics of cercariae was identified by histochemical examination and PCR method.Results The pictures of miracidia were recorded after co-cultivation for 1 h,12 h,24 h and 3 d.The shedding of the cilia of miracidia occurred within 12 h,parasites exhibited continuous stretching movements after

  11. Generation and analysis of 113 adult stage Schistosoma japonicum (Chinese strain) expressed sequence tags%113个日本血吸虫(中国大陆株)成虫表达序列标签的获取及分析

    Institute of Scientific and Technical Information of China (English)

    卞国武; 余新炳; 吴忠道

    2002-01-01

    Objective To rapidly and economically obtain knowledge about adult stage Schistosoma japonicum (Chinese strain) expressed genes using expressed sequence tag (EST) .Methods A directional cDNA library constructed from Schis tosoma japonicum (Chinese strain ) adult stage RNA was used to generate expressed sequence tags(ESTs).These were compared against an EMBL-parasites database and GENBANK database by BLASTn and BLASTx. Results A total of 314 phage clones were randomly selected for generating expressed sequence tags(ESTs). From these clones, 132 EST-qu ality sequence were obtained. Among these EST-quality sequences,113 ESTs were successfully submitted to the dbEST at GenBanK. A total of 7.6% of these EST- quality sequences were previously identified sequence of Schistosoma japonicum, while 4.5% were putatively identified sequences of Schistosoma japonicum. A total of 23.5% of these EST-quality sequences were putatively identified sequence of Schistosoma mansoni or other organisms. 57.6% had no matches in the database and were classified as unknown sequences. Most ESTs with the putative prote in identified belonged to housekeeping proteins. Information about several inte resting genes was found. Conclusion Partial cDNA sequencing to generate express ed sequence tags (ESTs) has the potential to rapidly and economically increase our knowledge about adult stage Schistosoma japonicum(Chinese strain) expressed genes.%目的运用表达序列标签(Expressed Sequence Tag, EST)技术快速、节约、有效地获得有关日本血吸虫(中国大陆株)成虫表达基因的知识.方法随机挑取日本血吸虫(中国大陆株)成虫cDNA 文库单个重组克隆进行部分测序以获得EST,获得的EST通过BLAST程序同EMBL寄生虫数据库和GeneBank数据库进行比较及同源性分析.结果本研究共随机挑取日本血吸虫(中国大陆株)成虫cDNA 文库单个重组克隆314个,获得了132个有EST价值的序列,其中113个成功在GeneBank db

  12. In vitro effects of EGCG on Schistosoma japonicum cercariae%表没食子儿茶素没食子酸酯(EGCG)体外抗日本血吸虫尾蚴作用的研究

    Institute of Scientific and Technical Information of China (English)

    冯金梅; 袁发浒; 刘锴; 张泽华; 胡松; 邱文洪; Stephen Hsu; 宋文剑

    2016-01-01

    日本血吸虫尾蚴具有体外杀伤作用,发生的形态结构可能影响其侵袭宿主皮肤的能力.%Objectives To investigate whether epigallocatechin-3-gallate (EGCG) kills Schistosoma japonicum cercariae in vitro and its effects on the ultrastructure of those cercariae in order to evaluate whether EGCG protects against infection with S.japonicum cercariae.Methods Cercariae of S.japonicum were placed in distilled water (control group) or dif ferent concentrations of EGCG (0.2% and 1 %).Cercariae were treated for 30 and 60 min and then observed with a dissecting microscope.Cercariae that were treated for 30 min were also observed using scanning electron microscopy.Male BALB/c mice,6-8 weeks of age,were randomly divided into tlree groups with six animals each:group Ⅰ (control group,infected with S.japonicurn cercariae treated with 0% EGCG for 30 min),group Ⅱ (infected with S.japonicum cercariae treated with 0.1 % EGCG for 30 min),and group Ⅲ (infected with S.japonicum cercariae treated with 1% EGCG for 30 min).All experimental mice were anesthetized and percutaneously infected with 30 cercariae of S.japonicum that were placed on shaved abdominal skin.The animals were maintained in conventional animal housing at the Labo ratory Animal Center of Jianghan University,China.All mice were given standard rodent food pellets and water ad libitum.At six weeks post-infection,the mice were sacrificed and adult worms of S.japonicum were collected,and the rate of infection,the average worm burden,and the rate of worm clearance were calculated.Results Under a dissecting microscope,S.japonicum cercariae treated with 0% EGCG for 30 min (the controls) had a normal morphology and they moved freely,while cercariae treated with 0% EGCG for 60 min were rigid,curled up into a ball,and not moving.Cercariae that were treated with 0.2 % or 1% EGCG for 30 min were highly active,their tails lashed rapidly,they twitched,and their bodies were deformed

  13. Evaluation of early diagnostic value of 6 antigens from Schistosoma japonicum in mice%血吸虫感染小鼠早期诊断抗原的研究

    Institute of Scientific and Technical Information of China (English)

    王玠; 余传信; 殷旭仁; 钱春艳; 宋丽君; 许永良; 何伟; 曹国群

    2011-01-01

    值,免疫印迹试验的敏感性比酶联免疫吸附试验高.%Objective To find out the candidate antigen for immunoreagent, which could be used to diagnose Schistosoma ja-ponicum infection early in mice. Methods The mice were infected with cereariae of S. japonicum Chinese mainland strain. The sera of mice before and after infection at different time were collected. The recombinant fusion protein (GST-HD) of the large hydro-philic domain (HD) of 23 kDa membrane protein of 5. japonicum with the Glutathione-S-transferase (GST) of S. japonicum, soluble eggs antigen (SEA), TSP2 hydrophilic domain of S. japonkum (TSP2HD), IL4-inducing principle of S. mansoni eggs (IPSE), fusion protein GST-SjMPIO (SjMP-10), and recombinant S. japonicum (Chinese strain) signaling protein 14-3-3 (Sj 14-3-3) were used as diagnostic antigens, the specific IgG and IgM antibodies were measured respectively by enzyme linked immunosorbent assay (ELI-SA). The antigens with the value of diagnosing schistosomiasis early were screened by analyzing the changes of the levels of specific IgG (or IgM) antibodies and the positive rates of specific antibodies in the sera of mice before and post infection at different time. Moreover, the antigen' s value of early diagnosis was further validated by Immunoblot. Results On the 18th, 21st and 28th day post infection, the positive rates of specific antibody IgM against GST-HD were 60%, 70% and 100%, respectively; the positive rates of specific antibody IgG against GST-HD were 40%, 60% and 90%, respectively. The positive rates of antibody IgM againstSEA were 50%, 60% and 90%, respectively; the positive rates of antibody IgG against SEA were 20%, 50% and 70%, respectively. The positive rates of IgM against TSP2HD were 30%, 40% and 50%, respectively; the rates of IgG against TSP2HD were 20%, 30% and 70%, respectively. The positive rates of IgM against IPSE were 20%, 30% and 50%, respectively; the positive rates of IgG against IPSE were 20%, 30% and 60

  14. Effect of a suspension of niclosamide on cercariae of Schistosoma japonicum and prevention of infections%氯硝柳胺悬浮剂灭蚴和预防血吸虫感染的研究

    Institute of Scientific and Technical Information of China (English)

    神学慧; 戴建荣; 李叶芳; 梁幼生; 李洪军; 唐建霞; 陈祥平

    2011-01-01

    Objective To investigate the effect of a suspension of niclosamide (SN) on cercariae of Schistosoma japonicum and effective concentrations of that suspension in order to provide scientific evidence for control strategies to prevent acute schistosomiasis. Methods SN was weighed and formulated into solutions with active concentrations of niclosamide of 1×10-5, 0. 5×10-5, 1×10-6, 0.5×10-6, 1× 10-1, and 0. 5×10-7 ml/100 ml in the lab, and the effect of SN on cercariae was determined. Solutions with an active concentration of niclosamide of 1. 2× 10-5, 1. 2× 10-6, 1. 2 ×10-7, 1.2×10-8, 1.2×10-0, and 1. 2×10-10 were formulated. Cercariae were counted 0, 10 and 30 min after 200 S. japonicum cercariae were transferred to the solutions and 30 min after mice were infected with the cercariae. Forty-five days after infection, the mice were dissected and the state of infection was observed. SN with a concentration of 0. 02 g/ m2 was sprayed for 1 h on the surface of water in the field, and then the infectivity of water was determined using sentinel mice. Results All cercariae were killed after coming into contact with an active concentration of niclosamide of more than 0. 5× 10-6 for 1 min, an active concentration of niclosamide of more than 0. 5× 10-7 for 2 min, or an active concentration of niclosamide of more than 1× 10-8 for 30 min. However, all survived after coming into contact with an active concentration of niclosamide of less than 0. 5 × 10-8 for under 60 min. After the cercariae came into contact with niclosamide of 1. 2 × 10-5 for 10 or 30 min, the cercariae failed to infect mice. When the cercariae were exposed to niclosamide of 1. 2 × 10-6 for 10 min, 80% of mice were infected, and the worm burden of mice was 3. 2 worms per mouse, but 30 min later the rate of infection in mice decreased to 0. Water in the field was infective, with a rate of infection in sentinel mice of 33. 33%. After 0. 02 g/m2 SN was sprayed on the water surface for 1 h, water

  15. 日本血吸虫26kDa谷胱甘肽S-转移酶基因真核表达载体的构建及序列测定%CONSTRUCTION AND SEQUENCING OF 26-KILODALTON GLUTATHIONE TRANSFERASES GENE OF SCHISTOSOMA JAPONICUM INTO EUKARYOTIC EXPRESSION VECTOR

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    目的扩增日本血吸虫26kDa谷胱甘肽S-转移酶(Sj26GST)基因片段,构建其重组真核表达载体,以研制SjGST核酸疫苗。方法根据已知基因序列设计一对引物,运用RT-PCR技术扩增Sj26GST目的基因片段,将其克隆到pcD-NA3质粒中,并经酶切、PCR鉴定,测序证实。结果获得GST-pcDNA3重组克隆。结论:本实验获得Sj26GST重组克隆,为进一步研制核酸疫苗创造了条件。%Aim To amplify the gene of 26-Kilodalton glutathione S-transferases of Schistosoma japonicum and construct a recombinant eukaryotic expression vector. Methods One pair of primers was designed according to the sequence of GST. The gene fragment of Sj26GST was obtained by RT-PCR,then it was cloned into a vector pcDNA3. It was corroborated through restriction enzymes cleaving,PCR and sequencing. Results GST-pcDNA3 recombinant clone was constructed successfully. Conclusions Sj26GST is a promising candidate of vaccine for Schistosoma japonicum. GSTrecombinant clone was achieved in the experiment. It is prepared for further study for DNA vaccine.

  16. Extraction of IgG from serum of rabbit infected with Schistosoma japonicum by saturated ammonium sulphate%硫酸铵法提取血吸虫感染兔血清IgG初探

    Institute of Scientific and Technical Information of China (English)

    兰炜明; 姜唯声; 谢曙英; 徐银; 戴坤教; 高晓晖; 曾小军

    2016-01-01

    Objective To explore the feasibility of preparing the schistosomiasis positive reference product by the method of ammonium sulfate precipitation to extract the IgG from the infected rabbits ’ serum of Schistosoma japonicum. Methods We used the saturated ammonium sulfate solution to precipitate and separate the IgG from the infected rabbits ’ serum. Compared the titer of infected rabbits’ serum with the separated IgG,and compared unfreeze-drying titer of IgG with the freeze-drying by IHA,also studied the stability of freeze-drying IgG after we dissolve with water. Results The titer of the infected rabbits’ serum was 1:320 used to extract IgG by the method of ammonium sulfate precipitation,we dialysised,concentrated the liquid of the extracted IgG to the original volume,the extracted IgG titer was 1:320. the negative rabbits’ serum and sucrose as the protein protective agent added to the extracted IgG to freeze-drying,the extracted IgG titer of the unfreeze-drying and the freeze-drying were 1:160. The titer of the freeze-drying IgG dissolved in water changed with the time,the 0th,3rd,6th and 8th titer is 1:160,1:80,1:40 and 1:40 respectively. Conclusion All of the process of the ammonium sulfate precipitation method to extract the IgG and the freeze-dry-ing can not chang the titer of the antibody,but the process adding the protein protective agent to the extracted IgG reduced the titer,the titer of freeze-drying IgG dissolve with water falled after the 3rd. The ammonium sulfate precipitation method is suitable for extracting the IgG from the infected rabbit serum.%目的:探索硫酸铵法提取血吸虫感染兔血清IgG用于制备阳性参考品的可行性。方法采用饱和硫酸铵溶液对感染兔血清中IgG进行沉淀分离,IHA试剂测定分离前后、冻干前后的抗体滴度以及冻干IgG复溶后的稳定性。结果滴度为1:320的血吸虫感染兔血清经硫酸铵沉淀法提取IgG,经透析、浓缩至

  17. An investigation of polymorphism in the tetraspanin-2 gene of Schistosoma japonicum%日本血吸虫被膜蛋白Sj-TSP2基因序列多态性分析

    Institute of Scientific and Technical Information of China (English)

    张丽丽; 徐新东; 潘卫庆

    2011-01-01

    Objective To identify the polymorphic sites of Schistosoma japonicum tegument protein tetraspanin-2 (Sj-TSP2).Methods Paired worms were separated in cold PBS with the help of forceps under a microscope.Total RNA of individual worms was extracted by Trizol reagent.First strands cDNAs were synthesized using reverse transcriptase.The open reading frame(ORF) of Sj-TSP2 was amplified by PCR.The resulting PCR products were sequenced after inserted into the pBlu2KSM vector between BamH I and EcoR I sites.DnaSP was used to calculate the alleles and the polymorphic sites.Mega 4 was used to build Neighborjoining tree and calculate the ratio of non-synonymous to synonymous substitutions (dN/dS).Results 46 fulllength ORFs of Sj-TSP2 were successfully amplified by RT-PCR from 60 individual worms.In total,92 fulllength sequences were obtained from 46 worms,containing 20 unique alleles.20 alleles were generated by point mutation and Sj-TSP2-10 was the predominant allele.There were a total of 85 polymorphism sites.Of the 63 resulting codon changes,27 gave rise to amino acid substitution (non-synonymous changes).Among them,22 amino acid substitutions were located in the extracellular loop 2.Conclusion Sj-TSP2 was an extensive polymorphic gene with 20 alleles identified.%目的 分析日本血吸虫Sj-TSP2序列多态性.方法 抽提单条日本血吸虫成虫的总RNA,逆转录合成cDNA,PCR扩增目的片段.将目的片段克隆到载体pBlue2KSM中转化大肠杆菌,挑取10个阳性克隆测序.将获得的Sj-TSP2序列导入DnaSP序列分析软件进行分析,计算等位基因数目,鉴定单核苷酸多态性位点(single nuclear polymorphism,SNP).同时将序列导入Mega 4序列分析软件,计算错义突变率与同义突变率的比值(dN/dS),进行进化分析,构建进化树.结果 共抽提了60条日本血吸虫成虫的总RNA,其中46条通过RT-PCR获得了目的基因Sj-TSP2;序列分析共鉴定了20个等位基因,主要是由点突变引起,Sj-P2-10是

  18. 重组日本血吸虫磷酸丙糖转移酶免疫保护性的研究%Preliminary study of recombinant the triosephosphate transferase in immuno-protection of Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    陈多玲; 张婧; 姚涌; 汪学龙

    2012-01-01

    Objective To observe the immuno-protection in mice attacked with cercaria for a new vaccine candidate of Schistosoma japonicum, the recombinant protein triosephosphate transferase (SjTPM). Methods The BALB/c mice were immunized with the recombinant SjTPM in the dose of 50μg/100μl、 100μg/100μl and 200μg/100μl, the saline solution was used for control group, then attacked with cercaria. The percentage of worm reduction and egg reduction was calculated to evaluate the immuno-protection effect. HE staining method was used to detect the hepatic changes and granulomatous changes in the liver tissue. Results The percentage of worm reduction was 27.1%,32.1% and 35.2%, respectively, in the experimental groups. The number of eggs in liver tissue was reduced by 34.2% ,39.29% ,43.66%, respectively. There were remarkable difference among the experimental groups and control group by t-test. There was no remarkable difference between the experimental groups by X2-test. Gross observation showed that the livers in control groups were dark brown and hard, with miliary nodules or worm eggs diffused in the livers. The recombinant SjTPM group has some extent of amelioration, the livers were bright red, relative soft and smooth, with less nodules of worm eggs in livers. Hepatic histological section demonstrated that there're generous large granulo- degeneration and necrosis of hepatic cells. The recombinant SjTPM vaccine group reversed the hepatic injury better than the control group. The structure of hepatic lobule was integrated on the whole, and the inflammatory reaction around the worm egg granulomas was light. Conclusion The molecular vaccine of rSjTPM had some immuno-protection effect.It suggested that rSjTPM was a potential candidate of S.japonicum.%目的 观察重组日本血吸虫磷酸丙糖转移酶(rSj-TPM)在小鼠抗血吸虫感染中的免疫保护作用,寻找新的血吸虫疫苗候选分子.方法 以rSj-TPM每鼠50μg/100μl、100μg/100μl和200μg/100μl

  19. Advances in Schistosoma japonicum recombinant protein glutathione-S-transferase(SjGST)and Schistosoma japonicum signaling protein 14-3-3(Sj14-3-3)%日本血吸虫GST蛋白和14-3-3蛋白的研究进展

    Institute of Scientific and Technical Information of China (English)

    刘文; 徐振山; 宋礼华

    2011-01-01

    Schistosomiasis is one of the serious health-threatening parasitic zoonoses to human beings. It has been presented in 76 countries and regions. More than 200 million people have been suffered from the infectious diseases all over the world. At present,some comprehensive precautionary and therapeutic measures were introduced to prevent schistosomiasis of human beings and animals simultaneously. The measures have brought some obvious positive effects. but the high cost combined with the high re-infection rate and such problems are still barriers to the application of the measures. Thus, to seek novel therapeutic medicines, candidates for the vaccines and also manage to develop standard immunologic diagnosis reagents with high specificity and sensitivity is the essential topic of the basic schistosomiasis research currently. The recent study of progress of the SjGST protein was reviewed, most potential candidates for the vaccines presented by WHO were summarized. Besides. this paper briefly reviewed the Sj14-3-3 protein which was involved in many functions of Schistosoma.%血吸虫病是严重危害人类健康的人兽共患病之一,在全球范围内,血吸虫病曾在76个国家和地区流行,有超过2亿人感染了血吸虫病.目前,世界上采取了一些人畜同步治疗等综合防治措施,虽取得了一定的成效,但仍面临着成本高、再感染率高等一系列问题.因此,寻找新的治疗药物、疫苗候选分子以及开发高度特异且敏感的标准化免疫诊断试剂是当前日本血吸虫病基础研究的重要内容.主要对WHO提出的最具潜力的疫苗候选分子血吸虫GST蛋白以及参与血吸虫许多生物学功能的14-3-3蛋白的最新研究进展进行一些阐述.

  20. 日本血吸虫感染不同相容性动物宿主的比较研究%Comparative study on Schistosoma japonicum infection in different permissive animal hosts

    Institute of Scientific and Technical Information of China (English)

    杨健美; 林矫矫; 苑纯秀; 冯新港; 傅志强; 石耀军; 刘金明; 洪炀; 李浩; 陆珂

    2012-01-01

    The aim was to understand the difference for worm development , worm morphology and host's histopatholo-gy in Schistosoma j aponicum (S. j aponicum) infected animal hosts , including natural hosts and experimental animal hosts . We artificially infected six animal hosts (yellow cattle , water buffalo , goat, New Zealand rabbit , BALB/c mice , and Wistar rat) with the same source of S . japonicum cercariae . The parasites were perfused through the hepatic portal vein on day 49 after infection. The male and female worms were detached manually and counted , and the length and width of the worms were measured . The results showed that the worm recoveries in permissive hosts were higher than that in non permissive hosts , and the length of the worms from permissive hosts were greater than those from non permissive hosts . Parasites in all the hosts could develop into maturation and cause liver damage in their hosts except for Wistar rat . The livers from permissive hosts were fulfilled with white egg nodules , and composed of egg- granulomas ; while the livers from water buffalo were red but with few egg nodules ; the livers from Wistar rat had no damage at all . The histologies! sections from livers of infected natural hosts showed that in yellow cattle and goat , hepatocytes displayed mild swelling , and a large number of inflammatory cells were seen to be in -filtrating and aggregating , including eosinophils and lymphocytes , and typical striped eosinophilic deposits were observed . Compared with yellow cattle and goat , the structural integrity of the hepatic lobules in infected buffalo was intacted in the liv -ers . There was actinomorphous distribution of hepatic cord centered on central veins , polygonal hepatocytes without edema and inflammatory cell infiltration , leaving only scattered neutrophils and monocytes . This study provides reference data for labora- tory and field studies in understanding the differences of S. j aponicum infection in different

  1. 日本血吸虫中国株次黄嘌呤鸟嘌呤磷酸核糖转移酶的克隆、表达及其免疫保护性研究%Cloning, Expression and Immunization of The Hypoxanthine-guanine Phosphoribosyltransferase for Schistosoma japonicum Chinese Strain

    Institute of Scientific and Technical Information of China (English)

    余俊龙; 吕志跃; 彭先楚; 周松华; 刘雪琴; 汪世平; 何卓; 戴橄; 李文凯; 姜孝新; 曾少华; 肖小芹; 徐绍锐

    2006-01-01

    A 1 270 bp full-length cDNA fragment was obtained from the Schistosoma japonicum (Chinese strain) adult cDNA library after the '3' and 5' ends of the incomplete expression sequence tag (EST) of hypoxanthine-guanine phosphoribosyltransferase of Schistosoma japonicum (SjHGPRT) were amplified by the anchored PCR with 2 pairs of primer that were designed according to the published incomplete SjHGPRT EST and the sequence of multiclone sites of library λgt1 1 vector. Sequence analysis indicated that this fragment, with an identity of 82% to hypoxanthine-guanine phosphoribosyltransferase ofSchistosoma mansoni (SmHGPRT), contained a complete open reading frame(ORF). The deduced amino acid sequence showed 83% identity to that of SmHGPRT. This fragment was cloned into the prokaryotic expression vector pQE30, and subsequently sequenced and expressed in Escherichia coli. SDS-PAGE revealed that M of the recombinant protein was about 28 ku. Western-blot analysis showed that the recombinant protein was recognized by the polyclonal antisera from rabbits immunized with Schistosoma japonicum adult worm antigen. Mice vaccinated with recombinant protein revealed significant worm burden, liver eggs per gram (LEPG), fecal eggs per gram (FEPG) and intrauterine eggs of the female worms reduction percentage, compared with the controls. Taken together, the SjHGPRT full-length cDNA can be cloned and expressed in E. coli as a recombinant protein that elicited immunity against the challenge infection with Schistosoma japonicum, indicating its potential as a partia1 protection vaccine candidate.%根据基因库中日本血吸虫次黄嘌呤鸟嘌呤磷酸核糖转移酶(HGPRT)EST(BU803192)以及日本血吸虫成虫cDNA文库载体λgt11多克隆位点邻近核苷酸序列设计引物,以日本血吸虫成虫cDNA文库为模板,采用锚式PCR对SjHGPRT基因不完整的3'端和5'端进行扩增、测序,用电子软件拼接,获得SjHGPRT全长cDNA(1 270 bp),经序列分析,推断该片段

  2. DNA polymorphism in the partial mitochondrial atp6 gene among Schistosoma japonicum isolates from Hunan Province%湖南省日本血吸虫线粒体atp6基因部分序列的多态性分析

    Institute of Scientific and Technical Information of China (English)

    夏英定; 汪世平; 李娟; 吴昌义; 田智; 尹铁球; 周云飞; 张树菊; 冯其梅

    2011-01-01

    目的 研究湖南省日本血吸虫不同地域自然隔离群线粒体ATP合成酶F0亚单位6基因(atp6)部分序列的遗传多态性,为湖南不同地域自然隔离群日本血吸虫的遗传特性研究提供实验依据.方法 采用试剂盒抽提基因组总DNA,以特异性引物对线粒体atp6基因进行PCR扩增,通过单链构象多态性技术(SSCP)筛选出差异带型并进行测序,用DNAStar 5.0及Mega 4.0软件进行比对分析.结果 湖南省5个流行区的日本血吸虫PCR扩增后获得了483 bp atp6部分序列,检测出17个变异位点,变异率为3.52%,雌雄虫之间的差异为0.0%~1.3%,不同地理来源虫株间的差异率为0.0%~1.5%.聚类分析结果表明:湖南不同地域自然隔离群日本血吸虫的种系发育不能按照其来源归为相应的系统发育进化树.结论 湖南不同地域自然隔离群日本血吸虫的atp6基因存在明显的个体差异,是否由于生存环境的选择压力造成了同一地理来源日本血吸虫个体之间atp6基因的遗传差异,其原因尚有待于进一步研究.%The aim of this study was to examinc sequence variation in the partial mitochondrial ATP synthase F0 subunit atp6 gene of Schistosoma japonicum and to provide foundation for studying the characterization and genetic structure of S. japonicum from different endemic origins of Hunan province. After extracting genomic DNA, atp6 was amplified with specific primers and screened by SSCP technique. Representative samples with variable banding patterns were selected for sequencing.DNAStar 5.0 and Mega 4.0 were used to analyze the sequences subsequently. Results showed that partial atp6 gene of S. japonicum was 483 bp, and 17 mutation sites were detected (3. 52%). Sequence variations for the partial atp6 were 0.0% -1.3% between male and female parasites, and 0.0%-1.5 % among isolates from different geographical origins. Phylogenetic analysis revealed that S. japonicum in Hunan province could not be

  3. Comparative evaluation of Schistosoma mansoni, Schistosoma intercalatum, and Schistosoma haematobium alkaline phosphatase antigenicity by the alkaline phosphatase immunoassay (APIA).

    Science.gov (United States)

    Cesari, I M; Ballén, D E; Mendoza, L; Ferrer, A; Pointier, J-P; Kombila, M; Richard-Lenoble, D; Théron, A

    2014-04-01

    To know if alkaline phosphatase (AP) from schistosomes other than Schistosoma mansoni can be used as diagnostic marker for schistosomiasis in alkaline phosphatase immunocapture assay (APIA), we comparatively tested n-butanol extracts of adult worm membranes from a Venezuelan (JL) strain of S. mansoni (Ven/AWBE/Sm); a Cameroonian (EDEN) strain of Schistosoma intercalatum (Cam/AWBE/Si) and a Yemeni strain of Schistosoma haematobium (Yem/AWBE/Sh). APIA was evaluated with sera of patients from Venezuela, Senegal, and Gabon infected with S. mansoni, from Gabon infected with S. intercalatum or S. haematobium, from Chine infected with Schistosoma japonicum and from Cambodian patients infected with Schistosoma mekongi. Results indicate that 92.5% (37/40) of Venezuela sera, 75% (15/20) of Senegal sera, 39.5% (17/43) of S. haematobium sera, and 19.2% (5/26) S. intercalatum sera were APIA-positive with the Ven/AWBE/Sm preparation. APIA with the Cam/AWBE/Si preparation showed that 53.8% of S. intercalatum-positive sera had anti-AP antibodies, and 51.2% S. haematobium-positive sera cross-immunocapturing the S. intercalatum AP. APIA performed with Yem/AWBE/Sh showed that 55.8% S. haematobium sera were positive. Only two out of nine S. japonicum sera were APIA-positive with the Ven/AWBE/Sm and Cam/AWBE/Si, and no reaction was observed with Cambodian S. mekongi-positive sera. AP activity was shown to be present in all the schistosome species/strains studied. The use of APIA as a tool to explore the APs antigenicity and the presence of Schistosoma sp. infections through the detection of anti-Schistosoma sp. AP antibodies in a host, allowed us to demonstrate the antigenicity of APs of S. mansoni, S. intercalatum, and S. haematobium.

  4. Immuno-protection of SJIR-2 DNA vaccine with microspheres adjuvant in mice challenged with Schistosoma japonicum%日本血吸虫SJIR-2纳米微球核酸疫苗的免疫保护性研究

    Institute of Scientific and Technical Information of China (English)

    王正印; 潘丽红; 汪学龙

    2016-01-01

    目的:研究日本血吸虫胰岛素受体-2( SJIR-2)纳米微球核酸疫苗对小鼠攻击感染的免疫保护效果。方法构建pEGFP-SJIR-2重组质粒,双酶切鉴定并测序,大量提取pEGFP-SJIR-2质粒,用壳聚糖( CHS)修饰的聚乳酸-羟基乙酸共聚物( PLGA)微球包裹,用包裹后的SJIR-2纳米微球免疫小鼠。将40只雌性BALB/c小鼠随机分为4组( n=10),分别注射 PBS、空 pEGFP 质粒、CHS-PLGA 微球和 CHS-PL-GA-pEGFP-SJIR-2微球各100μg免疫小鼠,末次免疫2周后,用日本血吸虫尾蚴攻击感染小鼠,每次免疫及感染尾蚴前收集各组小鼠血清,ELISA法检测各组小鼠血清内免疫球蛋白( IgG)水平的变化。小鼠感染尾蚴42 d后全部剖杀,收集成虫和虫卵并计算减虫率和减卵率。结果成功构建了pEGFP-SJIR-2重组质粒,与PBS组比较, CHS-PLGA-pEGFP-SJIR-2组的成虫数和虫卵数差异有统计学意义( P<0.01)。CHS-PLGA-pEGFP-SJIR-2组的减虫率和减卵率分别为37.36%和46.82%,和PBS组相比,CHS-PLGA-pEGFP-SJIR-2组小鼠血清内IgG水平比明显增高( P<0.01),而pEGFP组和CHS-PLGA组成虫数和虫卵数与PBS组比较差异无统计学意义。结论 SJIR-2纳米微球核酸疫苗对感染血吸虫的BALB/c小鼠有一定的免疫保护效果,对其潜在的候选抗原疫苗的价值尚需深入研究。%Objective To research the immuno-protection of SJIR-2 DNA vaccine with nanometer microspheres a-gainst Schistosoma japonicum infection in mice. Methods To construct eukaryotic expression plasmid pEGFP-SJIR-2, identified by double digestion and sequenced delivery. The recombinant plasmid pEGFP-SJIR-2 was ex-tracted and was encapsulated into PLGA nanometer microspheres which were modified by CHS. 40 female BALB/c mice were randomly divided into 4 groups (n=10), each group of mice were injected with PBS, empty pEGFP plasmid, CHS-PLGA nanometer microspheres and CHS-PLGA-pEGFP-SJIR-2 nanometer microspheres 100 μg, re-spectively. Two weeks

  5. Survival of Oncomelania Snails from Lake Regions and Mountain Regions and Their Susceptibility to Schistosoma Japonicum under Simulated Field Conditions%模拟自然条件下湖沼型和山区型钉螺存活率及其血吸虫易感性研究

    Institute of Scientific and Technical Information of China (English)

    龚珍强; 李传昌; 汪雁; 黄重峰; 刘大仁; 陶波; 周宪民; 邹节新

    2014-01-01

    Objective To compare the 4-month survive rates of ribbed shell Oncomelania snails from lake regions of Xingzi County of Jiangxi Province,ribbed shell Oncomelania snails from mountain regions of Yushan County of Jiangxi Province and smooth shell Oncomelania snails from mountain regions of Pengshan County of Sichuan Province and their susceptibility to Schis-tosoma japonicum under simulated natural conditions.Methods Lake-region ribbed shell On-comelania snails,mountain-region ribbed shell Oncomelania snails and mountain-region smooth shell Oncomelania snails were collected from Xingzi County of Jiangxi Province,Yushan County of Jiangxi Province and Pengshan County of Sichuan Province,respectively.The infected and un-infected snails were screened using the cercariae escaping method.The Schistosoma japonicum-in-fected snails from Xingzi County were used to infect a rabbit.The liver was removed and miracidia were obtained after 45 days.Then snails were infected with the miracidia at a ratio of 120 at 26-30 ℃ under fluorescent lamp irradiation.These infected snails were fed for 4 months in Schistoso-miasis Control Station of Xingzi County under simulated natural conditions,and the survival rate and infection rate were determined.Results Oncomelania snails from mountain regions of Yushan County of Jiangxi Province and mountain regions of Pengshan County of Sichuan Province could survive under simulated natural conditions in Xingzi County of Jiangxi Province.But their survival rates were significantly lower than those from lake regions of Xingzi County of Jiangxi Province (P < 0.001 ).Furthermore,Oncomelania snails from lake regions of Xingzi County of Jiangxi Province and mountain regions of Yushan County of Jiangxi Province,but not those from moun-tain regions of Pengshan County of Sichuan Province,were easily infected with Schistosoma ja-ponicum miracidia from Xingzi County of Jiangxi Province.In addition,the infection rate of snails from lake regions of Xingzi

  6. 日本血吸虫SIEA66-68kDa抗原的分离、纯化及免疫保护性研究%Isolation and purification of the 66-68 kDa soluble immature egg antigen Schistosoma japonicum and its protective immunity

    Institute of Scientific and Technical Information of China (English)

    姜孝新; 汪世平; 刘雪琴; 李庆华; 刘明社; 何卓; 彭先楚; 徐绍锐

    2011-01-01

    To investigate the immunoprotection of 66-68 kDa soluble antigen of immature egg of Schistosoma japonicum (SIEA66-68kDa) and to evaluate its feasibility of being used as a molecular vaccine antigen, the SIEA66-68kDa was prepared through SDS-PAGE electrophoresis, protein band resection, electro-elution and ultrafiltration, and the isolated product of protein was used as antigen to immunize Kunming strain of mice. After 3 courses of immunization, mice were challenged with cercariae of S. japonicum, and the reduction rates of worms and eggs were compared with those of the control group with inoculation of PBS. It was found that the mono-specific antibody response against SIEA66-68kDa protein could be induced in mice immunized with this protein. The SIEA66-68kDa could induce significant reduction in adult worms (41.70 %). The rates of egg reduction in the SIEA66-68kDa group reach 51.23%, 59.26% and 45.17% in the liver, intestine tissue and feces, respectively, with significant difference when compared with the control group. It is concluded that the 66-68 kDa immature egg antigen of S. japonicum could protect the Kunming strain of mice from the infection of this parasite, and it could be regarded as the target antigen for the preparation of vaccine against S. japonicum infection.%目的 研究日本血吸虫未成熟虫卵可溶性抗原66-68kDa (SIEA66-68kDa)的动物免疫保护力,并对其作为天然分子疫苗的可行性进行评估.方法 采用电泳切胶、电洗脱、超滤离心等技术,分离纯化SIEA66-68kDa天然分子抗原,免疫小鼠,待其产生抗体后进行尾蚴攻击感染(40尾/只),计算减虫率和减卵率.结果 SIEA66-68kDa天然蛋白质分子能刺激机体产生抗日本血吸虫的作用,其减虫率为41.70%,每克肝脏减卵率为51.23%,雌虫子宫减卵率为29.30%,每克大肠减卵率为59.26%,每克粪便减卵率为45.17%,与对照组相比差异具有统计学意义(P<0.05).结论 SIEA66-68kDa天然分子抗

  7. Bioinformatic analysis, cloning and expression of Schistosoma japonicum SCP/TAPS gene family%日本血吸虫SCP/TAPS基因家族的生物信息学分析和克隆表达

    Institute of Scientific and Technical Information of China (English)

    陈静芳; 胡旭初; 王乐旬; 余新炳

    2011-01-01

    目的 以黄蜂Vv Vesv5蛋白为检索源检索日本血吸虫EST数据库,对检索出的序列进行生物信息学分析,并挑选出有代表性的基因进行克隆表达,为日本血吸虫SCP/TAPS家族蛋白的功能研究打下一定的基础.方法 利用生物信息学软件对检索出的日本血吸虫SCP/TAPS家族蛋白进行生物信息学分析,经PCR确定后,以AAW24579为代表进行进一步的研究,将其命名为SjVAL2.根据SjVAL2的序列设计引物,通过RT-PCR扩增出全长编码区域,克隆到原核表达载体pET-28a(+)中,在大肠埃希菌中诱导表达并纯化,免疫印迹实验(Western-blotting)鉴定纯化的重组蛋白.结果 日本血吸虫SCP/TAPS家族中至少有17个不同的蛋白成员存在,多序列比对分析发现它们初步可以分为3个不同的亚组.以日本血吸虫成虫cDNA为模板RT-PCR扩增出SjVAL2序列,PCR、双酶切及DNA测序均证实pET-28a(+)-SjVAL2重组质粒构建成功.SDS-PAGE结果表明,目的基因在大肠埃希菌中获得高效表达,重组蛋白能识别日本血吸虫感染的小鼠血清,说明其具有免疫反应性.结论 日本血吸虫SCP/TAPS家族中至少有17个不同的蛋白成员存在,其中SjVAL2基因可在原核表达系统中高效表达,为进一步研究该家族蛋白的功能打下基础.%The Vespula vulgaris Wasp Venom Allergen 5 (Vv Vesv5, GenBank accession No. AAA30333) protein sequence was used in a tBLASTn search of S. Japonicum ESTs. Detail bioinformatic analysis were conducted and an interesting gene were cloned and expressed to investigate the role in the life cycle of S. Japonicum. After PCR confirmation of the sequences from parasite-derived cDNA, AAW24579 was selected for further study and named S. Japonicum Venom allergen-like transcript (SjVAL2). The coding region of SjVAL2 was amplified by RT-PCR using specific primers and cloned into prokary-otic expression vector pET-28a( +), then expressed in E. Coli BL2KDE3). The purified recombinant

  8. Schistosoma mansoni

    DEFF Research Database (Denmark)

    Friis, Henrik; Byskov, Jens

    1987-01-01

    Recent surveys in Ngamiland, Botswana, indicate increasing prevalence of Schistosoma mansoni infections. With the introduction of a schistosomiasis control programme, 354 of 373 schoolchildren were examined quantitatively for eggs of S. mansoni, and 317 were examined clinically for hepato...

  9. 日本血吸虫虫卵、童虫和雌雄成虫膜蛋白的双向电泳%Analysis of membrane proteins from egg, schistosomulum, adult male and female worm of Schistosoma japonicum by two dimensional electrophoresis

    Institute of Scientific and Technical Information of China (English)

    程国锋; 冯新港; 林矫矫; 石耀军; 陆柯; 周元聪; 蔡幼民

    2005-01-01

    Membrane proteins were extracted from eggs, schistosomulum, adult male and female worms of Schistosoma japonicum in order to analyze the differently expressed profile by two dimensional electrophoresis. Schistosomulum and adult worms were obtained from rabbits infected with 1 500 cercariaes on 14 and 42 days after challenge, respectively. Adult male and female worms on 42 days were manually detached and stored into liquid nitrogen until use. Eggs were collected by PercollTM from the liver of rabbits. ProteoPrep Membrane Extraction KitTM was employed to extracted membrane proteins by reducing and alkylating with TBP and iodoacetamide from 200mg of eggs, schistosomulums, adult male worm and female worms, respectively. Immobilized pH gradient strips with a linear pH range of 3-10(130mm) were rehydrated together with membrane proteins (30μg) in 250μl solution containing 7mol urea, 2mol thiourea, 2% SB3-10, 4% CHAPS, 40mmol Tris, 30mmol DTT, then separated on 12.5% SDS polyacrylamide gel for the second dimensional electrophoresis. Gels were stained with silver, scanned by Labscan, and analyzed using ImageMasterTM Analysis software. The 2D maps of egg, schistosomulum, adult female worm and male worm were showed 78±3, 67±3, 108±4 and 122±4 spots respectively. There were 35±1 spots which showed specific expression in female worm as compared with male worm, but 45±2 spots were in male worms. Most differently expressed spots between male and female worms were located in the area of 40-70kD and pI 4-7. The large number of unique spots from sehistosomulum was located in the area of alkalescence. The 2D map of for adult male worms uniquely showed 5 spots as compared with that of schistosomulum and female worm. The female worm showed 4 unique spots as compared with that of schistosomulum, egg and male worm. The unique spots between male and female worms were identified by the database of SWISS 2D-PAGE according to the molecular weight and isoelectronic point

  10. 日本血吸虫感染鼠巨噬细胞IL-13Rα2的表达%Expression on IL-13Rα2 in macrophages of Schistosoma japonicum-infected mice

    Institute of Scientific and Technical Information of China (English)

    王维; 郑胜生; 祁瑶; 闻惠琴; 刘丽丽; 沈继龙

    2011-01-01

    目的 检测IL-13的诱骗受体IL-13Rα2在血吸虫病巨噬细胞的表达,为从细胞水平探讨Th2型免疫性疾病分子病理机制奠定基础.方法 建立日本血吸虫病鼠模型,采用免疫荧光标记法分别检测肝肉芽肿和原代巨噬细胞IL-13Rα2蛋白的表达情况.结果 荧光显微镜下观察:肝肉芽肿内可见似"咖啡豆样"散在分布的IL-13Rα2免疫阳性信号,巨噬细胞膜内缘呈异常增强的环状荧光.结论 巨噬细胞是IL-13Rα2阳性表达细胞,IL-13Rα2是血吸虫病重要的免疫病理调节分子.%The purpose of this study was to investigate the expression on intracellular proteins of IL-13Rα2 in the macrophages of Schistosomiasis japonicum mice, and to offer the experiment foundation for elucidating IL-13 responding cell and mechanism of fibrogenesis in Th2-mediated diseases. A large number of mice were exposed to cercariae and established S. japonicum mouse model. IL-13Rα2 expressed in the granulomas of liver tissues and peritoneal macrophages was determined by immunofluorescent labeling with primary antibody of goat anti-mouse IL-13Rα2 antibody and monoclonal anti-CD68 antibody,respectively. Coffee bean-like IL-13Rα2 positive staining was showed in the hepatic granuloma in fluorescentmicroscopy. Co-expression of IL-13Rα2 and CD68 protein in peritoneal macrophages from 8-week infected mice was observed. It exhibited the distribution of IL-13Rα2 in cytoplasm concentrated on the inner layer of membrane in macrophages. It's suggested that IL-13Rα2 expressing macrophages might be a critical contributor to the immunopathogenesis of schistosomiasis.

  11. Cloning and function analysis of high mobility group box 1(HMGB1)protein of Schistosoma japonicum(Mainland strain)%日本血吸虫高速泳动家族B1蛋白基因的克隆表达与功能分析

    Institute of Scientific and Technical Information of China (English)

    姚媛; 许永良; 杨静; 余传信; 宋丽君; 殷旭仁; 王玠; 金一; 沈双; 张伟; 高玒

    2014-01-01

    Objective To clone and express a high mobility group box 1(HMGB1)protein of Schistosoma japonicum(Main-land strain)and analyze its function. Methods The DNA fragment of open reading frame encoding Sj HMGB1 protein was ampli-fied by RT-PCR from the mRNA of S. japonicum worms,then it was subcloned into the expression vector pET28a(+)to form the recombinant expression plasmid SjHMGB1-pET28a. The recombinant expression plasmid was transformed into the component E. coli BL21(DE3),and the tranformant containing recombinant expression plasmid was induced with IPTG to express the recombi-nant protein SjHMGB1. The recombinant SjHMGB1 protein was purified by affinity chromatography with nickel chelating affinity chromatography agarose gel. The Gel retard experiment and animal immunization were performed to analyze the DNA binding ca- pacity and the immunologic property of recombinant SjHMGB1. The expression levels of HMGB1 in different life cycle stages of S. japonicum were analyzed by Western bloting and RT-PCR. Female ICR mice were immunized with the recombinant SjHMGB1 pro-tein and infected with 45±2 cercariae of S. japonicum after three immunizations. Forty-two days post-infection,the worms and eggs of S. japonicum were recovered from the portal vein and liver tissue,respectively. The worm and egg reduction rates were calculat-ed respectively. Results A 530 bp of specific DNA fragment was amplified from mRNA of S. japonicum by RT-PCR,which was the open reading frame(ORF)encoding SjHMGB1protein confirmed by DNA sequencing analysis. The recombinant expression plasmid SjHMGB1-pET28a was constructed by cloning the ORF of SjHMGB1 into a expression vector pET28a(+). The bacterium transformants containing the recombinant plasmid expressed a soluble recombinant protein about 28 kDa after induced by IPTG, and the recombinant SjHMGB1 protein was purified by nickel chelating affinity chromatography. The gel retard experiment showed that the recombinant SjHMGB1 protein could bind

  12. 鄱阳湖湖北钉螺日本血吸虫和外睾吸虫尾蚴逸出模式及其流行病学的初步调查%Preliminary investigation on the prevalence and cercarial rhythm pattern of Schistosoma japonicum and Exorchis sp.in Oncomelania hupensis (Gastropoda) populations in marshlands of Poyang Lake, China

    Institute of Scientific and Technical Information of China (English)

    Yassir Sulieman; Theerakamol Pengsakul; 黄帅钦; 彭午弦

    2013-01-01

    Digenean trematodes are a widespread group of parasitic flatworms that use snails somewhat exclusively as their first intermediate host and one species can competitively exclude other species in a single individual snail. In this study, prevalence of the trematodes, Schistosoma japonicum and Exorchis sp., among populations of Oncomelania hupensis and population density of the snail in four different marshland locations around Poyang Lake, China, was studied during spring (April) and autumn (October) of 2012, by using a 0.11 m2 frame sampler. Also, the rhythm of cercarial liberation of the two trematodes from this area was investigated. Results showed that the prevalence of Exorchis sp. infection among O. hupensis populations was high (2.01%), compared to S. japonicum (0.15%) over the four investigated sites during the spring- and autumn-collected samples. The mean density of live snails in spring was 12.2 per 0.11 m2 with overall infection of 0.3 snail per 0.11 m2;while in autumn, the mean density of live snails was 6.7 per 0.11 m2 with infection rate of 0.2 snail per 0.11 m2. Cercarial release pattern from S. japonicum was circadian, with a single peak of emergence between 12:00 and 14:00 pm, whereas that of Exorchis sp. displayed a bimodal peak of emergence: an early emergence peak between 06:00 and 08:00 am, and a late emergence peak between 16:00 and 18:00 pm. Finaly, the higher prevalence of Exorchis sp. infection among O. hupensis populations compared to S. japonicum at different sites and seasons in this study necessitates further investigation concerning interaction among trematode infections within O. hupensis populations, especially in regard to Exorchis sp., due to its importance in schistosomiasis biological control.%目的 复殖吸虫是一种广泛分布的寄生性扁形动物,它们专一性地利用贝类作为第一中间宿主.自然环境中常见到一种贝类是数种吸虫的中间宿主,但该贝类的每一个体通常只存在一

  13. EXPRESSION OF SCHISTOSOMA JAPONICUM CALRETICULIN PROTEIN IN SF9 INSECT CELLS AND PRELIMINARY ANALYSIS OF ITS ROLE IN THE ACTIVATION OF DENDRITIC CELLS%日本血吸虫钙网织蛋白在Sf9昆虫细胞中的表达及其活化树突细胞的初步分析

    Institute of Scientific and Technical Information of China (English)

    李莹; 李运燕; 徐瑞敏; 苑纯秀; 杨健美; 林矫矫; 李利珍; 冯新港

    2012-01-01

    为获得在Sf9昆虫细胞中表达的日本血吸虫钙网织蛋白(Schistosoma japonicum calreticulin protein,SjCRT)并分析其活化小鼠骨髓来源树突状细胞的功能,将构建的重组杆状病毒转移载体pFastBacHTA-SjCRT转入DH 10Bac细胞,得到重组穿梭质粒reBacmid-SjCRT,再转染到Sf9昆虫细胞,进行重组蛋白的表达.用Westem blot和间接免疫荧光对表达蛋白进行鉴定.His柱亲和层析法纯化表达的蛋白,Westen blot鉴定纯化后的蛋白.从BALB/c小鼠产生骨髓来源的树突细胞mDCs,用纯化的重组SjCRT蛋白与mDCs共培养,流式细胞术检测mDCs细胞的表面分子MHCⅡ、CD40和CD86的表达.结果显示,在Sf9昆虫细胞中成功表达了SjCRT蛋白;纯化后的重组SjCRT蛋白既能被感染日本血吸虫42 d的兔阳性血清识别,也能被原核表达的重组SJC RT蛋白免疫鼠的血清所识别.流式细胞术结果显示,与对照组的相比,SjCRT蛋白刺激组mDCs细胞表面分子MHCⅡ和CD86的表达量显著增强(P<0.05).可见,在Sf9昆虫细胞中表达的SjCRT蛋白能刺激小鼠骨髓来源树突细胞表型的成熟.

  14. 蒿甲醚或吡喹酮早期治疗感染血吸虫尾蚴兔和犬的肝脏显微镜检查%Microscopic observations on livers of rabbits and dogs infected with Schistosoma japonicum cercariae and early treatment with artemether or praziquantel

    Institute of Scientific and Technical Information of China (English)

    肖树华; 杨元清; 张超威; 尤纪青

    1996-01-01

    To study the histopathological change of the liver of the hosts treated with artemether (Art) or praziquantel (Pra) in early stage after infection with Schistosoma japonicum cercariae. METHODS:Dogs infected once with schistosome cercariae were by the repeated dosing once every 1 or 2 wk for 2 -4 times. In rabbits, infected with 48 - 52 schistosome cercariae once every other day for 5 times,infection, followed by the repeated dosing every 1 or 2 wk for 2 - 3 times. RESULTS: After above mentioned dogs or rabbits were treated ig with Art,ArtC or Pra, the female worm reduction rates were 92.1% - 100 %. Histopathological examination showed that the reduction rates of total granuloma in the liver sections of the dogs and rabbits were 70.9 % - 97.3 % and 76.5 % - 97.4 %, respectively. Meantime, the structure of the hepatic lobules was normal with normal arrangement of the liver bundle. CoNCLusIoNS: Early treatment with Art or Pra exhibited a promising effect of protection of the liver of the dogs and rabbits infected with schistosome cercariae.%观察感染血吸虫尾蚴后早期用蒿甲醚(Art)或吡喹酮(Pra)治疗,对宿主肝组织的影响.方法:犬感染198-202条尾蚴后d 7ig Art 10 mg·kg-1,Art胶囊(ArtC)15 mg·kg-1或感染后d 21ig Pra 30-40 mg·kg-1,1-2 wk重复给药1次,共2-4次;兔每隔日感染48-52条尾蚴,共5次,并于第1次感染后d 7或d 21 ig 上述剂量的Art和Pra,停药后4-5 wk剖杀取肝作切片观察.结果:犬与兔经Art或Pra早期治疗后,减♀虫率达92.1%-100%,肝切片中的总虫卵肉芽肿数分别减少70.9%-97.3%和76.5%-97.4%,肝叶结构和肝索排列正常.结论:2种药物早期治疗对宿主肝脏有保护作用.

  15. 日本血吸虫TGF-β受体I基因胞外段的克隆与表达%Cloning and expression of extracelluar fragment of TGF-β receptor Ⅰ gene of Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    高志岩; 李孜; 黎小妍; 马长玲; 陈姗; 黄俊

    2011-01-01

    Objective To clone and express the extracelluar region ofTGF-β receptor-Ⅰ in Schistosomajaponicum(SjTβRIout),and to identify its immunologic functions. Methods Part of the coding region of the SjTβRI gene was obtained by rapid amplification of cDNA ends (RACE). The extracelluar fragment of SjTβRI gene was amplified by RT-PCR, cloned into the prokaryotic expression vector pET28a(+), and then confirmed by sequencing method. The plasmid was then transfected into E.coli and the expression was induced by IPTG. The recombinant protein was purified using NJ-ion affinity chromatography. SD rats were immunized with the purified protein and Freund's adjuvant. ELISA was used to detect the production of the SjTβRIout specific antibody. Results The pET28a(+)-SjTβRIout recombinant expression plasmid was successfully constructed. Sequence analysis showed that the SjTβRIout gene had 399 bp (encoding 133 amino acids) and the gene was highly homologous with Schistosoma mansoni TGF-β receptor Ⅰ. SjTβRIout recombinant protein with a molecular weight of 20 kDa was expressed in E. coli. The recombinant protein immunized SD rats produced a high titer (> 1:100 000 000) of antigen-specific IgG antibody.Conclusion The pET28a(+)-SjTβRIout recombinant protein was proved to be immunogenic.%目的 获得日本血吸虫TGF-β受体I胞外段(SjTβ3RIout)的序列,行原核克隆和表达,并分析其蛋白的免疫学功能.方法 应用cDNA末端快速扩增技术(rapid amplification of cDNA ends,RACE)方法获得SjTβRI基因cDNA部分序列,用RT-PCR方法扩增SjTβRI胞外段(SjTβRlout)基因,构建pET28a(+)重组原核表达载体,并测序鉴定.将质粒转入大肠埃希菌,IPTG诱导表达,并使用Ni离子亲和层析并透析处理获得纯化重组蛋白.使用纯化蛋白与弗氏佐剂共同免疫SD大鼠获得抗血清,使用ELISA方法检测血清中SjTBRIout特异抗体IgG含量.结果 成功构建SjTβRIout原核表达载体.测序鉴定显示SjTβ3RIout

  16. Investigation on the role of praziquantel against Schistosoma japonicum induced liver fibrosis and expression change of host miRNA%吡喹酮对日本血吸虫病肝纤维化的治疗作用及宿主miRNA表达变化

    Institute of Scientific and Technical Information of China (English)

    陈超; 白瑞璞; 何兴; 潘卫庆

    2015-01-01

    目的 研究吡喹酮对日本血吸虫虫卵引起的肝纤维化的治疗作用及其可能的机制. 方法 选用BALB/c小鼠建立日本血吸虫感染小鼠模型,实验组以250 mg/(kg·d)吡喹酮连续用药3d进行杀虫,再以600 mg/(kg·d)连续给药30 d进行抗纤维化治疗;对照组仅进行吡喹酮杀虫.以肝组织羟脯氨酸含量等指标评定小鼠肝纤维化的程度,并通过实时荧光定量PCR (real-time PCR)检测各组肝组织中某些微小RNA (microRNA,miRNA)的表达水平.各组检测结果以t检验进行统计学分析.结果 经过吡喹酮治疗后,小鼠肝脏羟脯氨酸含量等肝纤维化指标明显降低,小鼠肝脏羟脯氨酸含量:感染后第75天,感染组、杀虫组、治疗组依次为(0.86±0.07)、(0.66±0.06)、(0.25±0.05) mg/g肝湿重,治疗组低于感染组和杀虫组(t=12.86,P<0.01).同时小鼠肝脏的部分miRNA表达水平发生了明显改变,治疗组与杀虫组相比较,Col I、miR-223、miR-146b、miR-142-5p、miR-199a-5p、miR-34c*、miR-195依次下调了62%、38%、75%、77%、40%、54%和56%. 结论 吡喹酮对日本血吸虫病肝纤维化有一定的治疗作用,其作用可能与调节某些宿主miRNA的表达水平有关.%Objective To investigate the role and mechanism of praziquantel against Schistosoma japonicum egg induced liver fibrosis.Methods BALB/c mice were infected with Schistosoma japonicum as experimental mouse model.In the experimental group,praziquantel at 250 mg/(kg·d) for consecutive 3 days as insecticide treatment and at 600 mg/(kg·d) for 30 days of continuous administration as antifibrotic therapy while in the control group,the mice only given praziquantel insecticide treatment.The extent of liver fibrosis was evaluated by detection on liver hydroxyproline content and other indicators,and by detection on certain micro RNA (microRNA,miRNA) expression levels in liver tissues with real-time quantitative PC R (real-time PCR).Test results of each

  17. CONSTRUCTION OF EUKARYOTIC EXPRESSION PLASMID AND SEQUENCE ANALYSIS OF Sj16 GENE OF SCHISTOSOMA JAPONICUM CHINESE STRAIN%日本血吸虫(中国大陆株)Sj16基因真核表达载体的构建及序列分析

    Institute of Scientific and Technical Information of China (English)

    卞国武; 余新炳; 吴忠道; 徐劲; 单志新; 马长玲; 邵筱

    2001-01-01

    目的为了进一步研究日本血吸虫(中国大陆株)Sj16的免疫调节功能,丰富有关血吸虫免疫逃避知识.方法根据曼氏血吸虫Sm1 6基因已知序列设计合成一对引物,用PCR技术从日本血吸虫(中国大陆株)成虫cDNA文库中扩增Sj16基因;将Sj16基因定向克隆入pcDNA3,转化感受态DH5a菌;用酶切、PCR扩增鉴定筛选得到的重组阳性克隆.以重组pcDNA3-Sj16质粒为模板,用双脱氧链末端终止法测定Sj16基因序列,应用软件辅助分析Sj16序列及进行Sj16与曼氏血吸虫的Sm16同源性比较.结果从日本血吸虫(中国大陆株)成虫cDNA文库中获取Sj16基因,重组质粒中含有Sj16基因,成功构建日本血吸虫(中国大陆株)Sj16基因真核表达重组质粒pcDNA3-Sj16;Sj16基因开放读码框有354碱基,编码¨7氨基酸,N端18个氨基酸可能为信号肽序列;Sj16与Sm16有高度同源性,只存在一个氨基酸的差异.结论成功克隆了Sj16基因的真核表达载体,并测定、分析了Sj16基因序列,为深入研究Sj16的免疫调节功能,丰富有关血吸虫免疫逃避知识打下基础.%Objective To further study the immunodulatory function of Schistosoma japonicum Sj16 and more obtain theknowledge about the immune evasion of schistosome. Methods A couple of primers were designed according to theknown sequence of Schistosoma mansoni Sm16 and the Sj16 gene obtained by amplification from cDNA library of S. japon-icum Chinese strain by using PCR technique. By cloning Sj16 gene into a eukaryotic expression vector, pcDNA3, a recombi-nant pcDNA3-Sj16 was constructed and transferred into DH5α. The positive recombinant pcDNA3-Sj16 was screened and i-dentified by agarose gel electrophoresis, endonuclease digestion and PCR technique. The pcDNA3-Sj1 6 recombinant plasmidwas used as template and nucleotide sequence of Sj16 gene was determined by the dideoxy chain termination method. Usingsoftware to analyze the structure and sequence homology of Sj16

  18. Congenital Transmission of Schistosoma japonicumin the Rabbit

    Institute of Scientific and Technical Information of China (English)

    QianBao-zhen; H.O.Bogh; M.V.Johansen; WangPeng-peng

    2005-01-01

    Fourteen pregnant rabbits were each infected with 300 cercariae of Schistosoma japonicum and divided into two groups.Group M (n =8)was infected during mid-gestation (the organogenetic stage)and group L (n=6)was infected during late-gestation (the post-organogenetic stage).Mother rabbits and rabbit kittens were killed 45-60 days after infection and perfused in order to obtain worm counts.Furthermore,faecal egg counts and tissue egg counts from livers were obtained from the mother rabbits as well as the rabbit kittens.All mother rabbits became infected harbouring 207.6+20.2 and 220.0+27.5 adult worms in group M and L,respectively.In groups M and L, 13.5%and 46.7% of the kittens were infected,respectively,In 12 of 14 litters at least one kitten was infected.Tne infected kittens harboured between one and three adult S.japonicum.The livers of the kittens infected with a worm pair displaced lesions,as a result of egg deposition.The results, therefore,show that congenital transmission of S.japonicum can occur in rabbits.The close anatomical resemblance between the rabbit and human placenta may be indicative of the presence of congenital transmssion of S.japomcum infection in humans.

  19. Phenotype comparison between bone marrow derived dendritic cell and DC2.4 cell stimulated with antigen from Schistosoma japonicum%血吸虫抗原刺激小鼠骨髓来源的树突状细胞与DC2.4细胞的表型比较

    Institute of Scientific and Technical Information of China (English)

    李小红; 曹建平; 汤林华; 王胜军; 成静

    2011-01-01

    目的 研究比较小鼠树突状细胞DC2.4和骨髓来源树突状细胞(bone marrow derived dendritic cell,BMDC)经血吸虫抗原谷胱甘肽转移酶(GST)刺激后表面分子的表达异同.方法 骨髓来源的细胞经白介素4(interleukin 4,IL-4)、粒细胞-巨噬细胞集落刺激因子(granulocyte-macrophage colonystimulating factor,GM-CSF)诱导培养,获得树突状细胞.常规方法培养DC2.4.体外用日本血吸虫抗原GST刺激前述两种细胞,以PBS和脂多糖(lipopolysaccharide,LPS)作对照,流式细胞仪检测细胞表面分子CD40、CDSO、CD86的平均荧光强度,并进行统计学分析.结果 日本血吸虫抗原GST刺激BMDC后,表面分子CD40、CD80、CD86的平均荧光强度依次为100.39、42.38、170.83,与PBS对照组比较,CD40无明显变化,而CD80、CD86表达上调(P<0.01);GST刺激DC2.4后,细胞表面分子CD40、CD80、CD86的平均荧光强度依次为23.73、72.13、59.58,与PBS对照组比较,CD40和CD86表达上调(P<0.01),而CD80变化不明显.结论 DC2.4与BMDC经日本血吸虫抗原刺激后表面分子的表达变化不同.%Objective To compare the phenotypes of bone marrow derived dendritic cell(BMDC)and DC2.4 cell stimulated with GST from Schistosoma japonicum.Methods Bone marrow cells were cultured in the presence of IL-4 and GM-CSF to induce dendritic cells.DC2.4 cells were cultured as routine.Both cells were stimulated with GST and the expressions of CD40,CD80 and CD86 on the cells'surface were analyzed by FACS,using PBS and lipopolysaccharide as controls. Results After stimulating with GST,the means of fluorescence intensity(MFI)for CD40,CD80 and CD86 on BMDC surface were 100.39,42.38 and 170.83,respectively.Compared with PBS control,the MFI of CD80 and CD86 on BMDC,but not CD40,enhanced significantly.The MFIs of CD40.CD80 and CD86 on DC2.4 loaded by GST were 23.73,72.13 and 59.58 respectively.Compared with PBS control,the expressions of CD40 and CD86 enhanced significantly after schistosome

  20. Recombinant Plasmid ZLW/pEGFP-C2 Transfection into Schistosomula of Schistosoma japonicum%日本血吸虫表膜特异结合肽重组质粒ZLW/pEGFP-C2体外转染日本血吸虫童虫的研究

    Institute of Scientific and Technical Information of China (English)

    刘彦; 曾庆仁; 杨胜辉; 魏琦; 周军; 李立新; 兰玲梅

    2011-01-01

    目的 观察日本血吸虫表膜特异结合肽合成基因(ZLW)构建的ZLW/pEGFP-C2重组质粒体外转染H本血吸虫童虫的效率,了解其抗虫作用.方法 用0.75%二甲基亚砜(DMSO)和高浓度质粒浸泡法,使用重组质粒ZLW/pEGFP-C2转染机械转化的日本血吸虫脱尾童虫,以瞬时表达的增强型绿色荧光蛋白(EGFP)基因为报告基因,在倒置荧光镜下观察虫体.抽提转染后培养48 h虫体的总RNA和全虫蛋白,分别用RT-PCR和蛋白质印迹(Western blotting)检测ZLW基因和EGFP基因在虫体内的表达情况.于转染后培养24、48、72和96 h,用美蓝染色法鉴别虫体存活情况,并计数.以上试验均设空质粒组和Tris-HCl缓冲盐溶液(TBS)对照组.结果 镜下观察结果显示,转染率约为10%,其EGFP主要分布于虫体皮层,以口、腹吸盘最为明显;空质粒转染组虫体腹吸盘处略带绿色荧光.RT-PCR结果显示,ZLW/pEGFP-C2转染组虫体的扩增产物大小约为259 bp,测序结果与ZLW序列一致.Western blotting分析证实EGFP基因在童虫体内表达.抗虫效果显示,转染后24 h和48 h,ZLW/pEGFP-C2转染组童虫死亡率(14.0%,48.8%)与空质粒组(15.9%,45.7%)和TBS对照组(16.9%,50.3%)间的差异无统计学意义(P>0.05);转染后72 h,ZLW/pEGFP-C2转染组死亡率(92.7%)高于空质粒组(73.2%)和TBS对照组(76.8%)(P<0.01);转染后96 h,ZLW/pEGFP-C2转染组童虫死亡率达100%.结论 DMSO作用的高浓度质粒浸泡法可将重组质粒ZLW/pEGFP-C2引入日本血吸虫章虫体内,并获得表达.%Objective To study the efficiency of ZLW/pEGFP-C2 plasmid transfection into Schistosoma japonicum schistosomula and observe its in vitro effect of anti-schistosomula. Methods Recombinant plasmid ZLW/pEGFP-C2 was transfected into mechanically transformed schistosomula by immersion in 0.75% DMSO and high concentration of plasmid. Enhanced green fluorescent protein (EGFP) transfected cells were observed under inverted fluorescence

  1. 血吸虫病肝纤维化小鼠IL-22表达及肝星状细胞衰老的意义%Dynamic expressions of IL-22 and hepatic stellate cells senescence in mice in-fected with Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    李兰; 孙颖; 王洪武; 黄宇; 黄加权; 宁琴

    2014-01-01

    Objective To investigate the dynamic expressions of interleukin-22(IL-22),Interleukin-22 receptor 1(IL-22R1),and hepatic stellate cells(HSC)senescence in mice with Schistosoma japonicum infection. Methods A murine model of S. japonicum infection was established and the serum samples and liver tissues were collected 4,6,8,12 weeks post-infection. The serum samples were detected for the levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST). The pathological changes and proliferation of hepatic collagen fibers in the liver tissue were observed after HE staining and Masson staining. The HSC senescence in fibrotic livers was determined by the detection of senescence-associatedβ-galactosidase(SA-β-Gal). Sandwich ELISA was used to measure the expressions of IL-22,and Real-time PCR was used to test the mRNA levels of IL-22 and IL-22R1. The control group without S. japonicum infection was set up. Results The serum levels of ALT and AST signifi-cantly increased 8 weeks and 12 weeks after the infection(vs. 0 week,all P<0.05). The level of IL-22 increased 4 weeks and 6 weeks after the infection(vs. 0 week,both P<0.05),but reduced 8 weeks post-infection,and was even lower 12 weeks post-in-fection(vs. 4 weeks and 6 weeks,both P<0.01). Being consistent with the dynamic expression of IL-22 protein,the mRNA ex-pression of IL-22 began to increase 4 weeks and reached the peak 6 weeks after the infection(vs. 0 week,both P<0.05),and continuously declined 8 weeks and 12 weeks post-infections(vs. 6 weeks,both P<0.05). The increase of the expression of IL-22R1 mRNA was correlated with the progression of fibrosis,and the peak was in 12 weeks post-infections(vs. 0 week and 6 weeks,both P<0.05). The number of senescence-associated beta-galactosidase-positive HSCs was reduced with the decreasing expression of IL-22 in the advanced liver fibrosis. Conclusion IL-22 and IL-22R1 are involved in the pathogenesis of schistoso-miasis liver fibrosis. As an inflammation factor

  2. 日本血吸虫重组质粒pGEX-Sj26GST的构建及在大肠埃希菌BL21(DE3)中的表达%Construction of a recombinant plasmid pGEX-Sj26GST of Schistosoma japonicum and expression in Escherichia coli BL21 (DE3)

    Institute of Scientific and Technical Information of China (English)

    张丽; 李文桂; 向进平

    2013-01-01

    目的 构建日本血吸虫(Schistosoma japonic um,Sj)重组质粒pGEX-Sj26GST,并研究其在大肠埃希菌(Escheric hia coli,E.coli BL21 (DE3)中的表达.方法 使用RNeasy Mini试剂盒提取sj成虫总RNA,实时荧光定量PCR(real-time PCR,RT-PCR)扩增日本血吸虫26 kDa的谷胱甘肽-S-转移酶(Sj26GST)编码基因,并将扩增产物定向克隆到pGEX-1λT载体上,构建重组质粒pGEX-Sj26GST;将重组质粒电穿孔转化感受态大肠埃希菌BL21(DE3),异丙基硫代-β-D-半乳糖苷(IPTG)诱导其表达;通过十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分析表达产物及蛋白印迹(Western blot)试验鉴定表达的融合蛋白的抗原性.结果 RT-PCR扩增出676 bp大小的Sj26GST抗原编码基因;双酶切试验证实Sj26GST抗原编码基因成功克隆到pGEX-1λT载体中;SDS-PAGE分析出现相对分子质量(Mr)约52×103的融合蛋白,薄层扫描分析显示表达的融合蛋白约占总菌体蛋白的20%,Western blot结果显示融合蛋白可被Sj感染的兔血清识别.结论 成功构建重组质粒pGEX-Sj26GST,其在大肠埃希菌BL21(DE3)中可获得表达,且表达的融合蛋白具有特异的抗原性.%Objective To construct and express a recombinant plasmid pGEX-Sj26GST of Schistosoma japonicum(Sj) in Escherichia coli(E.coli) BL21 (DE3).Methods Total RNA was extracted from Sj adult worms by RNeasy Mini kit,26 kilodalton glutathione-S-transferases of Schistosomajaponicum (Sj26GST) antigen gene was amplified by real-time PCR(RT-PCR) from the total RNA,then cloned into a prokaryotic expression plasmid pGEX1λT and transformed into E.coli BL21 (DE3) to construct pGEX-Sj26GST; BL21 (pGEX-Sj26GST) was induced with isopropyl-beta-D-thiogalactopyranoside (IPTG),and the expressed products were analyzed and identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE) and Western blotting.Results The 676 bp Sj26GST gene was successfully amplified by RT-PCR and restriction enzyme double

  3. Studies on the effect and mechanism of CD4+ CD25+ regulatory T calls in Schistosoma japonicum immune evasion%CD4+CD25+调节性T细胞在血吸虫免疫逃避中的作用及机制研究

    Institute of Scientific and Technical Information of China (English)

    唐春莲; 郭思洁; 杨进; 祝青; 刘晓宏

    2014-01-01

    (IL-5) and interleukin-10 (IL-10) in cultural suspension of splenic cells was detected by sandwich-ELISA after stimulation with ConA.Results Worm burden in anti-CD25 mAb group (23.17 ± 6.94) was extremely lower than in infected control group [(30.17 ± 5.85) pg/mL,P =0.047].Compared to those of normal [(1.98 ±0.33),P =0.049] and infected control group [(2.68 ±0.12),P =0.000],the ration of CD4+ CD25+ Tregs were extremely low in anti-CD25 mAb group (1.28 ± 0.30).The level of IFN-γ in anti-CD25 mAb group [(386.87 ± 24.85) pg/mL,P =0.000] was extremely higher than that of infected control group (61.32 ± 8.75)pg/mL,while the levels of IL-4,IL-5 and IL-10 were not significant different among the three groups.Conclusion These results hint that CD4+ CD25+ Tregs contribute to the escape of S.japonicum from the host immune assault,while anti-CD25 mAb can partially block CD4+ CD25+ Tregs and favor the host to clear parasite by enhancing Th1-type immune response.

  4. Preliminary Observation on the Effect of Mefolquine against Egg Granuloma Formation in the Liver of Mice Infected with Schistosoma japonicum%甲氟喹对感染小鼠日本血吸虫虫卵肉芽肿形成影响的初步观察

    Institute of Scientific and Technical Information of China (English)

    肖树华; 张超威

    2013-01-01

    形成具有抑制作用.%Objective To explore whether mefolquine possesses the effect on granuloma formation induced by Schistosoma japonicum eggs. Methods Seventeen out of twenty-eight mice infected with 20 S. japonicum cercariae for 35 days were treated orally with mefloquine at a single dose of 200 mg/kg, and groups of 2-3 mice were sacrificed at various intervals post-treatment. The livers removed from each group of mice were fixed in 10% formaldhyde. While the remained 11 untreated mice divided into 6 groups (1-2 mice per group) were sacrificed at the same time periods as groups of mice treated with mefloquine, and their livers served as untreated corresponding controls. The granulomas with single egg in the center were counted and their diameters were measured using an ocular micrometer. The liver tissue sections were stained with hematoxylin and eosin (HE), Foot's or Mallory's methods for observation on histopathological alteration of egg granulomas, and on the appearance of reticular and collagen fibers within the granulomas. Results After infected mice were treated with mefloquine for 3, 7, 14, 21, 28 and 35 days, i.e., 38, 42, 49, 56, 63, and 70 days post-infection, the mean diameters of granuloma with single egg measured in the liver tissues section were (161 ±19), (175±13), (195±9), (171±40), (180±13), and (145±25)μm, respectively, and each of them was significantly lower than that of its corresponding control group of (189±18), (197±11), (211±12), (208±19), (203±16), and (207±36) μm (P<0.01 or P<0.05). Histopatological observation showed that in mice treated with mefloquine, the eosinophil-predominant inflammatory cells around the egg granuloma were sustained to 14-21 d post treatment (49-56 d post infection), which was significantly different from the corresponding control groups that all the eggs were surrounded by fibroblasts at 42 d post infection. Up to 28-35 d post treatment (63-70 d post infection), the boundary of egg granulomas distributed in the liver tissues of

  5. 日本血吸虫间接血凝法抗体检测试剂盒稀释液的改进%Alternative diluent used for the commercial IHA assay kit for detection of Schistosoma japonicum anti-body

    Institute of Scientific and Technical Information of China (English)

    章乐生; 王恩木; 张世清; 汪奇志; 呼明闯; 司武敏; 杨荣笙

    2014-01-01

    目的:确定一种适用于安徽安吉医药科技有限公司新包装试剂盒的稀释液,替代原包装中的冻干致敏红细胞稀释液(生理盐水)和血清标本稀释液(蒸馏水)。方法以pH7.2磷酸盐缓冲液作为稀释液制备新包装试剂盒替代原包装,比较新包装试剂盒与原包装的敏感性和特异性以及与肝吸虫、肺吸虫、姜片虫和钩虫等病人血清交叉反应情况;比较各试剂盒的最低检出量,观察凝集反应模式。结果新包装试剂盒与原包装敏感性分别为95%和94.2%,差异无统计学意义(χ2=0.000,P=1.000);特异性分别为96.9%和95.8%,差异无统计学意义(χ2=0.000,P=1.000);约登指数分别为0.92和0.9;最低检出量均为1:1280,磷酸盐缓冲液组凝集模式较好;与肝吸虫、姜片虫和钩虫病人的血清交叉反应率均为0,与肺吸虫的交叉反应率均为19.4%。结论确定磷酸盐缓冲液作为日本血吸虫间接血凝法抗体检测试剂盒新包装的稀释液,替代原包装中的致敏红细胞稀释液和标本稀释液。%Objective To evaluate the feasibility of a new diluent as a substitute for the original ones (sa⁃line used for diluting the lyophilized sensitized red cells, and distilled water for diluting the serum specimen) packaged in the commercial indirect hemagglutination (IHA)assay kit manufactured by Anhui Anji Pharmaceu⁃tical Technology Co., Ltd. Methods Phosphate buffered solution(PBS, pH7.2)was used to replace the diluent in the original kit, and two preparations were compared regarding the sensitivity and specificity in detection of Schistosoma japonicum as well as the cross-reactivity with liver fluke, lung fluke, giant intestinal fluke and hook⁃worm. The minimal quantification and the agglutination pattern in each group were also observed. Results The sensitivity and specificity were 95% and 96.9% for the modified kit, 94.2% and 95.8% for the original one

  6. The complete mitochondrial genomes of Schistosoma haematobium and Schistosoma spindale and the evolutionary history of mitochondrial genome changes among parasitic flatworms.

    Science.gov (United States)

    Littlewood, D Timothy J; Lockyer, Anne E; Webster, Bonnie L; Johnston, David A; Le, Thanh Hoa

    2006-05-01

    Complete mitochondrial genome sequences for the schistosomes Schistosoma haematobium and Schistosoma. spindale have been characterized. S. haematobium is the causative agent of urinary schistosomiasis in humans and S. spindale uses ruminants as its definitive host; both are transmitted by freshwater snail intermediate hosts. Results confirm a major gene order rearrangement among schistosomes in all traditional Schistosoma species groups other than Schistosoma japonicum; i.e., species groups S. mansoni, S. haematobium, and S. indicum. These data lend support to the 'out of Asia' (East and Southeast Asia) hypothesis for Schistosoma. The gene order change involves translocation of atp6-nad2-trnA and a rearrangement of nad3-nad1 relative to other parasitic flatworm mt genomes so far sequenced. Gene order and tRNA secondary structure changes (loss and acquisition of the DHU and/or TPsiC arms of trnC, trnF, and trnR) between mitochondrial genomes of these and other (digenean and cestode) flatworms were inferred by character mapping onto a phylogeny estimated from nuclear small subunit rRNA gene sequences of these same species, in order to find additional rare genomic changes suitable as synapomorphies. Denser and wider taxon sampling of mt genomes across the Platyhelminthes will validate these putative characters.

  7. Symbiotic efficiency of selected indigenous Bradyrhizobium japonicum strains

    Directory of Open Access Journals (Sweden)

    Branka Komesarović

    2007-12-01

    Full Text Available Symbiotic nitrogen fixation has a unique role in a sustainable agriculture because the utilization of this natural process enables significant reduction of mineral nitrogen fertilization. Thereby, selection of highly efficient symbiotic nitrogen fixators is of great importance for successful legume inoculation due to the fact that the strains of nodule bacteria strongly differ in their symbiotic efficiency, competitiveness and compatibility. The main aim of the present study was to evaluate the symbiotic characteristics of indigenous and reference Bradyrhizobium japonicum and Bradyrhizobium elkanii strains and their compatibility with different soybean cultivars in agroecological conditions of eastern Slavonia. During two years of investigations, twofactorial field trials were set up in a randomized complete block design with five replicates. The results obtained in this study show that all B. japonicum strains had positive effect on all analyzed properties and particularly on seed yield as the most important parameter in soybean production. The highest values for nodule dry weight were obtained by application of indigenous B.japonicum strain S69 that was isolated from the similar agroecologicalconditions. Although the application of B. elkanii strain S76T resulted in the most abundant nodulation, at the same time it induced chlorosis on leaves in full blossom stage. Regarding the tested soybean cultivars, Ika and Nada, significant differences in nodulation were determined. Considerable rainfall deficiency in 2002 had strong influence on all tested properties. The highest nitrogen content in soybean aerial parts as well as the seed yield were determined in application of indigenous B. japonicum strains S69 and S75. The obtained results suggest that these two indigenous strains can becharacterized as the most efficient ones and could be used for soybean inoculant production for that region.

  8. Clonning and sequence analysis of the 26 kDa glutathiones-transferase gene of Schistosoma mekongi.

    Science.gov (United States)

    Vichasri-Grams, S; Grams, R; Korge, G; Viyanant, V; Upatham, E S

    1997-09-01

    The number of genomic DNA or cDNA sequences of Schistosoma mekongi accessible in Genbank or EMBL is very limited up to now. Recently, two reports have appeared on the molecular phylogeny of Schistosoma species inferred from partial sequence data of rRNA genes; no further sequence data of S. mekongi is available yet. Knowledge of the molecular structure of protein coding genes of S. mekongi will provide a better understanding of gene function in the genus Schistosoma. A cDNA library of S. mekongi adult male was constructed and a cDNA encoding the 26 kDa glutathione S-transferase protein of this species was cloned. Sequence analysis of this cDNA confirmed the close phylogenetic relationship of S. mekongi to S. japonicum.

  9. 日本血吸虫重组质粒pGEX-Sj14-3-3-Sj32的构建及其在大肠埃希菌BL21(DE3)中的表达%Construction and expression of a recombinant plasmid pGEX-Sj14-3-3-Sj32 of Schistosoma japonicum in Escherichia coli BL21(DE3)

    Institute of Scientific and Technical Information of China (English)

    覃婷; 李文桂; 谭建蓉

    2015-01-01

    目的 构建日本血吸虫重组质粒pGEX-Sj14-3-3-Sj32,探讨重组质粒在大肠埃希菌BL21 (DE3)中的表达.方法 以重庆医科大学附属第一医院传染病寄生虫病研究所保存的质粒pGEX-Sj14-3-3和pET28α-Sj32为模板,通过PCR扩增Sj14-3-3和Sj32抗原编码基因,然后采用基因拼接法(gene SOEing)剪接Sj14-3-3和Sj32基因,得到Sj14-3-3-Sj32融合基因,定向克隆入穿梭载体pGEX-1 λT,构建重组质粒pGEX-Sj14-3-3-Sj32,并采用双酶切法进行验证.将重组质粒转化大肠埃希菌BL21 (DE3),经异丙基硫代-β-D-半乳糖苷(IPTG)诱导表达后,用十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和蛋白质免疫印迹(Western blot)法对表达产物进行分析鉴定.结果 基因拼接法得到约1 750 bp的Sj14-3-3-Sj32融合基因;双酶切证实Sj14-3-3-Sj32融合基因成功插入pGEX-1λT载体中;SDS-PAGE分析显示,表达产物为相对分子质量约为73×103的重组蛋白;Western blot显示,重组蛋白可被日本血吸虫感染的兔血清识别.结论 成功构建日本血吸虫重组质粒pGEX-Sj 14-3-3-Sj32,该重组质粒在大肠埃希菌BL21(DE3)中得到了高效融合表达,且表达的融合蛋白具有特异的抗原性.%Objective To construct and express a recombinant plasmid pGEX-Sj 14-3-3-Sj32 of Schistosoma japonicum in Escherichia coli (E.Coli) BL21 (DE3).Methods Sj14-3-3 and Sj32 antigen genes were amplified by PCR from template of plasmids pGEX-Sj14-3-3 and pET28α-Sj32 which were extracted from recombinant bacteria BL21 (pET28α-Sj32) and BL21 (pGEX-Sj14-3-3) stored in Institute of Infectious and Parasitic Disease of the First Affiliated Hospital of Chongqing Medical University.Sj14-3-3-Sj32 fusion gene obtained with gene SOEing was cloned into the vector pGEX-1λT to construct pGEX-Sj14-3-3-Sj32 which was identified by double digestion.The recombinant plasmid pGEX-Sj 14-3-3-Sj32 was transformed into E.Coli BL21 (DE3).The recombinant strains were induced by

  10. Characterization of a Partially Protective B-cell Epitope within the 62 kDa Antigen of Schistosoma japonicun

    Institute of Scientific and Technical Information of China (English)

    Lei ZHANG; Xue YANG; Yanfen YANG; Jiaqing ZHAO; Jianghua YANG; Feng LIU; Zhaosong ZHANG; Guanling WU; Chuan SU

    2007-01-01

    Approximately 200 million people worldwide currently suffer from schistosomiasis, one of the most important human parasitic diseases. Although an established infection can be treated with anthelminthics and praziquantel, vaccination would be the ideal method for integral control of schistosomiasis. Schistosoma mansoni IrⅤ-5, recommended as a vaccine candidate by the World Health Organization/Special Programme for Research and Training in Tropical Diseases, produced high protection in animal models. We therefore focused on its homolog, the Schistosoma japonicum 62 kDa antigen, and analyzed it using B cell/antibodyrelated databases and analysis tools for the prediction of B-cell epitopes. Epitope B3 was selected for further investigation. Experiments using a murine model indicated that mice immunized with B3 resulted in lymphocytes proliferation and produced high levels of specific immunoglobulin G and G1, but did not produce impressive cytokines. The vaccination showed partial protective immunity, measured by worm burden and anti-fecundity immunity against S. japonicum. These results indicated that the epitope B3 from S. japonicum 62-kDa antigen might act as a candidate immunogen for future epitope vaccine investigation.

  11. A novel chalcone derivative from Onychium japonicum

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    A novel chalcone derivative was isolated from Onychium japonicum and its structure was elucidated by spectroscopic methods.The isolated compound showed cytotoxic activity against the human carcinoma cell lines Hela and BEL-7402 in vitro.(C) 2007 Hong Quan Duan. Published by Elsevier B.V. on behalf of Chinese Chemical Society. All rights reserved.

  12. Development and evaluation of a sensitive PCR-ELISA system for detection of schistosoma infection in feces.

    Directory of Open Access Journals (Sweden)

    Luciana Inácia Gomes

    Full Text Available BACKGROUND: A PCR-enzyme-linked immunosorbent assay (PCR-ELISA was developed to overcome the need for sensitive techniques for the efficient diagnosis of Schistosoma infection in endemic settings with low parasitic burden. METHODOLOGY/PRINCIPAL FINDINGS: This system amplifies a 121-base pair tandem repeat DNA sequence, immobilizes the resultant 5' biotinylated product on streptavidin-coated strip-well microplates and uses anti-fluorescein antibodies conjugated to horseradish peroxidase to detect the hybridized fluorescein-labeled oligonucleotide probe. The detection limit of the Schistosoma PCR-ELISA system was determined to be 1.3 fg of S. mansoni genomic DNA (less than the amount found in a single cell and estimated to be 0.15 S. mansoni eggs per gram of feces (fractions of an egg. The system showed good precision and genus specificity since the DNA target was found in seven Schistosoma DNA samples: S. mansoni, S. haematobium, S. bovis, S. intercalatum, S. japonicum, S. magrebowiei and S. rhodaini. By evaluating 206 patients living in an endemic area in Brazil, the prevalence of S. mansoni infection was determined to be 18% by examining 12 Kato-Katz slides (41.7 mg/smear, 500 mg total of a single fecal sample from each person, while the Schistosoma PCR-ELISA identified a 30% rate of infection using 500-mg of the same fecal sample. When considering the Kato-Katz method as the reference test, artificial sensitivity and specificity rates of the PCR-ELISA system were 97.4% and 85.1%, respectively. The potential for estimating parasitic load by DNA detection in feces was assessed by comparing absorbance values and eggs per gram of feces, with a Spearman correlation coefficient of 0.700 (P<0.0001. CONCLUSIONS/SIGNIFICANCE: This study reports the development and field evaluation of a sensitive Schistosoma PCR-ELISA, a system that may serve as an alternative for diagnosing Schistosoma infection.

  13. 埃及血吸虫的发现及其生物学%Discovery and biology of Schistosoma haematobium

    Institute of Scientific and Technical Information of China (English)

    汪伟; 洪青标; 梁幼生

    2014-01-01

    血吸虫病是一种严重危害人类健康、影响社会经济发展的被忽略的热带病。寄生于人体的血吸虫主要有日本血吸虫(Schistosoma japonicum)、曼氏血吸虫(S. mansoni)、埃及血吸虫(S. haematobium)、间插血吸虫(S. intercalatum)和湄公血吸虫(S. mekongi)5种。作为最早发现的一种血吸虫(1852年),人体埃及血吸虫感染与膀胱癌发病密切相关,但相关生物学资料甚少。本文主要综述了埃及血吸虫的发现历程及其形态、生活史和生态。%Schistosomiasis is a neglected tropical disease that severely threatens human health and affects the socioeconomic development. The causative agent that parasitizes in humans mainly involves Schistosoma japonicum,S. mansoni,S. haematobi-um,S. intercalatum and S. mekongi. As the firstly identified schistosome,S. haematobium infection is found to strongly correlate with bladder cancer. This paper mainly reviews the discovery,morphology and life cycle of S. haematobium.

  14. Establishment of Experimental Model of Hepatic Schistosoma Japonicum Egg Granulomas in Mice

    Institute of Scientific and Technical Information of China (English)

    冯振卿; 李玉华; 仇镇宁; 薛婉芬; 管晓虹

    1998-01-01

    IntroductionSchistosomaegggranulomasaretheprinci-palpathologicbasisofschistosomiasis.Toes-tablishanappropriateexperimentalmod...

  15. Schistosoma japonicum:establishment of a mouse model that demonstrates concomitant immunity

    Institute of Scientific and Technical Information of China (English)

    HE Li 何立; YI Xin-yuan 易新元; JIANG Ming-sen 蒋明森; ZENG Xian-fang 曾宪芳; OUYANG Li 欧阳理; Larry McReynolds

    2004-01-01

    @@ Concomitant immunity to schistosome reinfection in vivo is generally thought to be the immune response induced by and relying on living adult worms.1,2 However, a few authors3,4 have suggested that the mechanism of resistance may be related to the hepatic pathology caused by the primary infection. The vascular pathology might enable the challenge schistosomula to escape from the hepatic portal system, thus preventing normal parasite sequestration and maturation in the liver.

  16. Molybdate transport by Bradyrhizobium japonicum bacteroids.

    OpenAIRE

    Maier, R J; Graham, L.

    1988-01-01

    Bacteroid suspensions of Bradyrhizobium japonicum USDA 136 isolated from soybeans grown in Mo-deficient conditions were able to transport molybdate at a nearly constant rate for up to 1 min. The apparent Km for molybdate was 0.1 microM, and the Vmax was about 5 pmol/min per mg (dry weight) of bacteroid. Supplementation of bacteroid suspensions with oxidizable carbon sources did not markedly increase molybdate uptake rates. Anaerobically isolated bacteroids accumulated twice as much Mo in 1 h ...

  17. Comparison of Schistosoma mansoni soluble cercarial antigens and soluble egg antigens for serodiagnosing schistosome infections.

    Directory of Open Access Journals (Sweden)

    Huw Smith

    Full Text Available A Schistosoma mansoni cercarial antigen preparation (cercarial transformation fluid--SmCTF was evaluated for detection of anti-schistosome antibodies in human sera in 4 collaborating laboratories. The performance of SmCTF was compared with that of S. mansoni egg antigens (SmSEA in an indirect enzyme-immunoassay (ELISA antigen assay, the latter being used routinely in 3 of the 4 participating laboratories to diagnose S. mansoni and S. haematobium infections. In the fourth laboratory the performance of SmCTF was compared with that of S. japonicum egg antigens (SjSEA in ELISA for detection of anti-S. japonicum antibodies. In all 4 laboratories the results given by SmCTF in ELISA were very similar to those given by the antigen preparation routinely used in the respective laboratory to detect anti-schistosome antibodies in human infection sera. In so far as the ELISA results from SmCTF are thus so little different from those given by schistosome egg antigens and also cheaper to produce, the former is a potentially useful new diagnostic aid for schistosomiasis.

  18. Schistosoma mansoni and Host-Parasite Interactions

    NARCIS (Netherlands)

    S.M-C.A. de Walick (Saskia)

    2015-01-01

    markdownabstract__Abstract__ Blood-dwelling parasitic trematodes (flatworms) of the genus Schistosoma cause the disease schistosomiasis or Bilharzia. There are 5 different Schistosoma species that infect humans and many other infecting different mammals. Over 200 million people worldwide are infect

  19. [Historical aspects of the risk factors of Schistosoma intercalatum schistosomiasis].

    Science.gov (United States)

    Jusot, J F; Simarro, P; De Muynck, A

    1996-01-01

    Bilharziosis is a considerable public health problem. It is caused by many species of schistosoma, four of which have wide geographical distribution: Schistosoma mansoni, S. haematobium, S. japonicum and S. intercalatum. The recently discovered S. intercalatum is limited to central and west Africa. Its spread is progressive and its pathogenicity is not completely known. S. intercalatum bilharziosis is usually manifested in the form of dysentery. The physiopathologic explanation of this clinical manifestation is less clear. Immunopathologically, the formation of an inflammatory granuloma constitutes the origin of its symptoms. This is due to many biological factors including delayed hypersensitivity reactions. All cellular immunity changes will facilitate the appearance of symptoms. Our aim has been to show the importance of malnutrition as a pathogenic factor of S. intercalatum bilharziosis. The initial research hypothesis was as follows: malnutrition plays a role in the evolution of a patient from an asymptomatic state of infection to a symptomatic state of illness. We carried out the study in the suburbs of Bata, in Equatorial Guinea. The inhabitants of Ncolombong, essentially rural immigrants, comprised our study population. Following their consent, we recruited individuals less than 45 years of age who had not taken praziquantel during the last 12 months. We included a total of 297 patients. Our study was a case-control, matching on sex and age. A case was defined as an infected patient with acute or chronic diarrhea occurring within the last month' preceding the stool sample analysis. All cases were retained after exhaustive screening of the study population. Each case (group 1) was matched with one or several asymptomatic infected patients (group 2) and two or several asymptomatic noninfected patients chosen at random (group 3). The definition of malnutrition was as follows: weight/height endemic area (OR = 0.14, CI 95%: 0.03-0.76) and a poor or average

  20. A link between arabinose utilization and oxalotrophy in Bradyrhizobium japonicum.

    Science.gov (United States)

    Koch, Marion; Delmotte, Nathanaël; Ahrens, Christian H; Omasits, Ulrich; Schneider, Kathrin; Danza, Francesco; Padhi, Barnali; Murset, Valérie; Braissant, Olivier; Vorholt, Julia A; Hennecke, Hauke; Pessi, Gabriella

    2014-04-01

    Rhizobia have a versatile catabolism that allows them to compete successfully with other microorganisms for nutrients in the soil and in the rhizosphere of their respective host plants. In this study, Bradyrhizobium japonicum USDA 110 was found to be able to utilize oxalate as the sole carbon source. A proteome analysis of cells grown in minimal medium containing arabinose suggested that oxalate oxidation extends the arabinose degradation branch via glycolaldehyde. A mutant of the key pathway genes oxc (for oxalyl-coenzyme A decarboxylase) and frc (for formyl-coenzyme A transferase) was constructed and shown to be (i) impaired in growth on arabinose and (ii) unable to grow on oxalate. Oxalate was detected in roots and, at elevated levels, in root nodules of four different B. japonicum host plants. Mixed-inoculation experiments with wild-type and oxc-frc mutant cells revealed that oxalotrophy might be a beneficial trait of B. japonicum at some stage during legume root nodule colonization.

  1. Organic Acid Metabolism by Isolated Rhizobium japonicum Bacteroids

    Science.gov (United States)

    Stovall, Iris; Cole, Michael

    1978-01-01

    Rhizobium japonicum bacteroids isolated from soybean (Glycine max L.) nodules oxidized 14C-labeled succinate, pyruvate, and acetate in a manner consistent with operation of the tricarboxylic acid cycle and a partial glyoxylate cycle. Substrate carbon was incorporated into all major cellular components (cell wall + membrane, nucleic acids, and protein). PMID:16660386

  2. [Content of rare earth elements in wild Hypericum japonicum Thunb].

    Science.gov (United States)

    Wei, Zhen-Lin; Rui, Yu-Kui; Tian, Zhi-Huan

    2009-06-01

    Rare earth elements are important nutritional elements for human health, and today more and more attention has been paid to the effective components in Chinese traditional medicine, especially to rare earth elements. Fifteen rare earth elements in wild hypericum japonicum Thunb were analyzed by the methods of ICP-MS. The results showed that the concentrations of La, Ce, Pr, Nd, Sm, Eu, Gd, Tb, Dy, Ho, Er, Yb, Tm, Lu and Y ranged from 6 ng x g(-1) x DW to 14 522 ng x g(-1) x DW, and among them the concentrations of La, Ce and Nd were higher than 2 000 ng x g(-1) x DW. Compared with the concentration of rare earth elements in rice, corn, wheat and barley, the total concentration of rare earth elements in hypericum japonicum Thunb was much higher, which could be the mechanism of curative effect of hypericum japonicum Thunb on liverish diseases. The character of elements and the content of rare earth elements in soil should be responsible for the difference, but the distributive mechanism of rare earth elements in hypericum japonicum Thunb should be further studied.

  3. Immunolocalization of Schistosoma mansoni and Schistosoma haematobium antigens reacting with their Egyptian snail vectors.

    Science.gov (United States)

    El-Dafrawy, Shadia M; Mohamed, Amira H; Hammam, Olfat A; Rabia, Ibrahim

    2007-12-01

    The reaction of the haemolymph and the tissue of infected intermediate hosts, Biomphalaria alexandrina and Bulinus truncatus to Schistosoma mansoni and S. haematobium antigens were investigated using the indirect immunoperoxidase technique. A new technique, Agarose cell block was used in collection of haemolymph which helped in collecting plenty of well formed cells in comparison to the ordinary one using the cytospin. Collected haemolymph and prepared tissues of uninfected and infected B. alexandria and B. truncatus were fixed and then reacted with anti-S. mansoni and anti-S. haematobium IgG polyclonal antibodies. The haemolymph and tissue of infected B. alexandrina and B. truncatus gave a positive peroxidase reaction represented by a brown colour. In haemolymph, the positive peroxidase reaction was detected mainly in the cytoplasm of the amoebocytes. In the tissue, it was detected in epithelial cells lining the tubules, male cells in the lumen of the tubules and in female oogonia cells along the periphery of the tubules. The similarity in the strength and distribution of positive reaction in B. alexandrina and B. truncates was observed as compared to control. Thus, the immunoperoxidase technique proved to be an effective indicator for the schistosome-antigen in the snails.

  4. SmCL3, a gastrodermal cysteine protease of the human blood fluke Schistosoma mansoni.

    Directory of Open Access Journals (Sweden)

    Jan Dvorák

    Full Text Available Blood flukes of the genus Schistosoma are platyhelminth parasites that infect 200 million people worldwide. Digestion of nutrients from the host bloodstream is essential for parasite development and reproduction. A network of proteolytic enzymes (proteases facilitates hydrolysis of host hemoglobin and serum proteins.We identified a new cathepsin L termed SmCL3 using PCR strategies based on S. mansoni EST sequence data. An ortholog is present in Schistosoma japonicum. SmCL3 was heterologously expressed as an active enzyme in the yeast, Pichia pastoris. Recombinant SmCL3 has a broad pH activity range against peptidyl substrates and is inhibited by Clan CA protease inhibitors. Consistent with a function in degrading host proteins, SmCL3 hydrolyzes serum albumin and hemoglobin, is localized to the adult gastrodermis, and is expressed mainly in those life stages infecting the mammalian host. The predominant form of SmCL3 in the parasite exists as a zymogen, which is unusual for proteases. This zymogen includes an unusually long prodomain with alpha helical secondary structure motifs. The striking specificity of SmCL3 for amino acids with large aromatic side chains (Trp and Tyr at the P2 substrate position, as determined with positional scanning-synthetic combinatorial library, is consistent with a molecular model that shows a large and deep S2 pocket. A sequence similarity network (SSN view clusters SmCL3 and other cathepsins L in accordance with previous large-scale phylogenetic analyses that identify six super kingdoms.SmCL3 is a gut-associated cathepsin L that may contribute to the network of proteases involved in degrading host blood proteins as nutrients. Furthermore, this enzyme exhibits some unusual sequence and biophysical features that may result in additional functions. The visualization of network inter-relationships among cathepsins L suggests that these enzymes are suitable 'marker sequences' for inclusion in future phylogenetic analyses.

  5. Hypericum japonicum Thunb. ex Murray: Phytochemistry, Pharmacology, Quality Control and Pharmacokinetics of an Important Herbal Medicine

    Directory of Open Access Journals (Sweden)

    Lin-Sheng Liu

    2014-07-01

    Full Text Available Hypericum japonicum Thunb. ex Murray is mainly distributed throughout Asia, Oceania and North America and is used as an important herbal medicine. H. japonicum contains many valuable secondary metabolites, such as flavonoids, phloroglucinols and xanthones and has hepatoprotective, anti-tumor, antibacterial, antiviral, and antioxidant activities and effects on the cardiovascular system and immunity. Coupled with phytochemical and pharmacological research, a series of analytical methods have been developed to evaluate the quality of H. japonicum based on its bioactive components. A pharmacokinetics study involved the absorption of two main flavonoids of H. japonicum in rats. This review aims to present an up-to-date and comprehensive overview of the phytochemistry, pharmacology, quality control and pharmacokinetics of H. japonicum, which should be useful for the greater development of H. japonicum, especially in the development of new drugs and therapeutics for various diseases.

  6. Detection of Extracellular Enzyme Activities in Ganoderma neo-japonicum

    OpenAIRE

    Jo, Woo-Sik; Park, Ha-Na; Cho, Doo-Hyun; Yoo, Young-Bok; Park, Seung-Chun

    2011-01-01

    The ability of Ganoderma to produce extracellular enzymes, including β-glucosidase, cellulase, avicelase, pectinase, xylanase, protease, amylase, and ligninase was tested in chromogenic media. β-glucosidase showed the highest activity, among the eight tested enzymes. In particular, Ganoderma neo-japonicum showed significantly stronger activity for β-glucosidase than that of the other enzymes. Two Ganoderma lucidum isolates showed moderate activity for avicelase; however, Ganoderma neo-japonic...

  7. Exposure versus Susceptibility as Alternative Bases for New Approaches to Surveillance for Schistosoma japonicum in Low Transmission Environments.

    Directory of Open Access Journals (Sweden)

    Shuo Wang

    2016-03-01

    Full Text Available Currently, schistosomiasis in China provides an excellent example of many of the challenges of moving from low transmission to the elimination of transmission for infectious diseases generally. In response to the surveillance dimension of these challenges, we here explore two strategic approaches to inform priorities for the development of improved methods addressed specifically to schistosomiasis in the low transmission environment. We utilize an individually-based model and the exposure data used earlier to explore surveillance strategies, one focused on exposure assessment and the second on our estimates of variability in individual susceptibility in the practical context of the current situation in China and the theoretical context of the behavior of transmission dynamics near the zero state. Our findings suggest that individual susceptibility is the major single determinant of infection intensity in both the low and medium risk environments. We conclude that there is considerable motivation to search for a biomarker of susceptibility to infection in humans, but that there would also be value in a method for monitoring surface waters for the free-swimming forms of the parasite in endemic or formerly endemic environments as an early warning of infection risk.

  8. Formation and Controlled Drug Release Using a Three-Component Supramolecular Hydrogel for Anti-Schistosoma Japonicum Cercariae

    Directory of Open Access Journals (Sweden)

    Yibao Li

    2016-03-01

    Full Text Available A novel three-component supramolecular hydrogel based on riboflavin, melamine and amino acid derivatives were constructed for controlled release of pesticides, Niclosamide derivatives. The formation of hydrogel may be attributed to self-assemble via hydrogen bonding and π–π interaction, which have been researched via scanning electron microscopy (SEM and Fourier transform infrared (FT-IR spectra. The rheological experiments showed that the hydrogel materials and drug-loaded hydrogel all demonstrated good mechanical strength and high stability. Further experimental results indicated that the drug-loaded hydrogels show large drug loadings, long-term release time and relatively higher efficiency to anti-cercariae in the water environment.

  9. NCBI nr-aa BLAST: CBRC-TTRU-01-1326 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-1326 emb|CAX75571.1| Transmembrane protein 118 [Schistosoma japonicum]... emb|CAX75572.1| Transmembrane protein 118 [Schistosoma japonicum] emb|CAX75574.1| Transmembrane protein 118 [Schistosoma... japonicum] emb|CAX75575.1| Transmembrane protein 118 [Schistosoma japonicum] CAX75571.1 0.36 27% ...

  10. Gene : CBRC-CJAC-01-0715 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available protein [Schistosoma japonicum] 3e-09 24% MRCYKYSTNTIPHYTDTLLLILIWCYKYSTNTIPHYTDTLLILILIWCYKYSTNTIPHYTETLLLILMRCYKYSTNT...IPYYTDTLLLILIWCYKYSTNTIPHYTDTLLILILIWCYKYSTNTIPHYTDTLLLILIRCYKYSTNTIPCYTETLLLLLYGATNTLLILYHTILIALLLILIWCYKYSTILXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXILY ...

  11. NCBI nr-aa BLAST: CBRC-MMUR-01-0199 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUR-01-0199 emb|CAX76110.1| putative tubulin, beta, 2 [Schistosoma japonicum]... emb|CAX76111.1| putative tubulin, beta, 2 [Schistosoma japonicum] emb|CAX76112.1| putative tubulin, beta, 2 [Schistosoma... japonicum] emb|CAX76113.1| putative tubulin, beta, 2 [Schistosoma japonicum] emb|CAX76114.1| p...utative tubulin, beta, 2 [Schistosoma japonicum] emb|CAX76115.1| putative tubulin, beta, 2 [Schistosoma... japonicum] emb|CAX76116.1| putative tubulin, beta, 2 [Schistosoma japonicum] CAX76110.1 2e-31 82% ...

  12. Susceptibility of Iraqi fresh water snails to infection with Schistosoma haematobium and Schistosoma mansoni Egyptian strains.

    Science.gov (United States)

    Wajdi, N A; Hussain, W I; El-Hawary, M F

    1979-01-01

    A great number of Egyptian workers and farmers are seeking settlement in Iraq and some of them proved to have either Schistosoma Haematobium (S.h.) or Schistosoma mansoni (S.m) or even mixed infection. Besides, there is the possibility that some of the Iraqi fresh water snails may prove to be susceptible to infection by one or both of the Schistosoma Egyptian strains. The present study deals with investigations on the susceptibility of Iraqi B. truncatus, Gyranaulus ehrenbergi, Physa c.f. fontinalis, Lymnea lagetis, Melanoides tuberculata and Melanopsis nodes by these parasites. Egyptian S. haematobium but not Egyptian S. mansoni infect Iraqi B. truncatus and both proved to be unable to infect any of the other snails included in the study. Yet, the number of cercariae shedded by B. truncatus snails infected with the Egyptian S. haematobium strain, was much less that the number of cercariae shedded by these snails when infected with the Iraqi S. Haematobium strain.

  13. The cercarial glycocalyx of Schistosoma mansoni

    OpenAIRE

    1985-01-01

    Cercariae, the freshwater stage of Schistosoma mansoni infectious to man, are covered by a single unit membrane and an immunogenic glycocalyx. When cercariae penetrate the host skin, they transform to schistosomula by shedding tails, secreting mucous and enzymes, and forming microvilli over their surface. Here the loss of the glycocalyx from cercariae transforming in vitro was studied morphologically and biochemically. By scanning electron microscopy, the glycocalyx was a dense mesh composed ...

  14. Role of Bradyrhizobium japonicum and Trichoderma spp. in the control of root rot disease of soybean

    Directory of Open Access Journals (Sweden)

    Syed Ehteshamul-Haque

    2014-08-01

    Full Text Available Seed treatment of soybean with Bndyrhizobium japonicum, Trichoderma harzianum, T. viride, T. hamatum, T. koningii and T. pseudokoningii significantly controlled the infection of 30-day-old seedlingsby Maerophomina phaseolina, Rhizoctonia solani and Fusarium spp. In 60-day-old plants Trichoderma spp.. and B. japonicum inhibited the grouth of R. solani and Fusarium spp., whereas the use of B. japonicum (TAL-102 with T. harzianum. T. viride, T. koningii and T. pseudokoningii controlled the infection by M. phaseolina. Greater grain yield was recorded when B. japonium (TAI-102 was used with T. hamatum.

  15. Soybean Lectin Enhances Biofilm Formation by Bradyrhizobium japonicum in the Absence of Plants

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    Julieta Pérez-Giménez

    2009-01-01

    Full Text Available Soybean lectin (SBL purified from soybean seeds by affinity chromatography strongly bound to Bradyrhizobium japonicum USDA 110 cell surface. This lectin enhanced biofilm formation by B. japonicum in a concentration-dependent manner. Presence of galactose during biofilm formation had different effects in the presence or absence of SBL. Biofilms were completely inhibited in the presence of both SBL and galactose, while in the absence of SBL, galactose was less inhibitory. SBL was very stable, since its agglutinating activity of B. japonicum cells as well as of human group A+ erythrocytes was resistant to preincubation for one week at 60°C. Hence, we propose that plant remnants might constitute a source of this lectin, which might remain active in soil and thus favor B. japonicum biofilm formation in the interval between soybean crop seasons.

  16. Germline transgenesis and insertional mutagenesis in Schistosoma mansoni mediated by murine leukemia virus.

    Directory of Open Access Journals (Sweden)

    Gabriel Rinaldi

    Full Text Available Functional studies will facilitate characterization of role and essentiality of newly available genome sequences of the human schistosomes, Schistosoma mansoni, S. japonicum and S. haematobium. To develop transgenesis as a functional approach for these pathogens, we previously demonstrated that pseudotyped murine leukemia virus (MLV can transduce schistosomes leading to chromosomal integration of reporter transgenes and short hairpin RNA cassettes. Here we investigated vertical transmission of transgenes through the developmental cycle of S. mansoni after introducing transgenes into eggs. Although MLV infection of schistosome eggs from mouse livers was efficient in terms of snail infectivity, >10-fold higher transgene copy numbers were detected in cercariae derived from in vitro laid eggs (IVLE. After infecting snails with miracidia from eggs transduced by MLV, sequencing of genomic DNA from cercariae released from the snails also revealed the presence of transgenes, demonstrating that transgenes had been transmitted through the asexual developmental cycle, and thereby confirming germline transgenesis. High-throughput sequencing of genomic DNA from schistosome populations exposed to MLV mapped widespread and random insertion of transgenes throughout the genome, along each of the autosomes and sex chromosomes, validating the utility of this approach for insertional mutagenesis. In addition, the germline-transmitted transgene encoding neomycin phosphotransferase rescued cultured schistosomules from toxicity of the antibiotic G418, and PCR analysis of eggs resulting from sexual reproduction of the transgenic worms in mice confirmed that retroviral transgenes were transmitted to the next (F1 generation. These findings provide the first description of wide-scale, random insertional mutagenesis of chromosomes and of germline transmission of a transgene in schistosomes. Transgenic lines of schistosomes expressing antibiotic resistance could advance

  17. Modeling and analysis of Schistosoma Argonaute protein molecular spatial conformation

    Directory of Open Access Journals (Sweden)

    Jianhua Zhang

    2011-08-01

    Conclusions: The information relationship between the structure and function of the Argonaute protein can be initially established with bioinformatics tools and the internet server, and this provides the theoretical basis for further clarifying the function of Schistosoma Argonaute protein.

  18. Gynecological manifestations, histopathological findings, and schistosoma-specific polymerase chain reaction results among women with Schistosoma haematobium infection

    DEFF Research Database (Denmark)

    Randrianasolo, Bodo Sahondra; Jourdan, Peter Mark; Ravoniarimbinina, Pascaline;

    2015-01-01

    BACKGROUND: The pathophysiology of female genital schistosomiasis (FGS) is only partially understood. This study aims to describe the histopathological findings, polymerase chain reaction (PCR) results, and gynecological manifestations of FGS in women with different intensities of Schistosoma...

  19. Crystal Structure of Schistosoma mansoni Adenosine Phosphorylase/5’-Methylthioadenosine Phosphorylase and Its Importance on Adenosine Salvage Pathway

    Science.gov (United States)

    Torini, Juliana Roberta; Brandão-Neto, José; DeMarco, Ricardo; Pereira, Humberto D'Muniz

    2016-01-01

    Schistosoma mansoni do not have de novo purine pathways and rely on purine salvage for their purine supply. It has been demonstrated that, unlike humans, the S. mansoni is able to produce adenine directly from adenosine, although the enzyme responsible for this activity was unknown. In the present work we show that S. mansoni 5´-deoxy-5´-methylthioadenosine phosphorylase (MTAP, E.C. 2.4.2.28) is capable of use adenosine as a substrate to the production of adenine. Through kinetics assays, we show that the Schistosoma mansoni MTAP (SmMTAP), unlike the mammalian MTAP, uses adenosine substrate with the same efficiency as MTA phosphorolysis, which suggests that this enzyme is part of the purine pathway salvage in S. mansoni and could be a promising target for anti-schistosoma therapies. Here, we present 13 SmMTAP structures from the wild type (WT), including three single and one double mutant, and generate a solid structural framework for structure description. These crystal structures of SmMTAP reveal that the active site contains three substitutions within and near the active site when compared to it mammalian counterpart, thus opening up the possibility of developing specific inhibitors to the parasite MTAP. The structural and kinetic data for 5 substrates reveal the structural basis for this interaction, providing substract for inteligent design of new compounds for block this enzyme activity. PMID:27935959

  20. Ganoderma neo-japonicum Imazeki revisited: Domestication study and antioxidant properties of its basidiocarps and mycelia.

    Science.gov (United States)

    Tan, Wee-Cheat; Kuppusamy, Umah Rani; Phan, Chia-Wei; Tan, Yee-Shin; Raman, Jegadeesh; Anuar, Azliza Mad; Sabaratnam, Vikineswary

    2015-07-27

    Mushroom cultivation benefits humankind as it deliberately encourages wild mushrooms to be commercially propagated while recycling agricultural wastes. Ganoderma neo-japonicum is a rare polypore mushroom found growing on decaying Schizostachyum brachycladium (a tropical bamboo) clumps in Malaysia. The Malaysian indigenous tribes including the Temuans and Temiars use the basidiocarps of G. neo-japonicum to treat various ailments including diabetes. In this study, the domestication of G. neo-japonicum in artificial logs of different agricultural residues was investigated. Sawdust promoted the mycelia spawn colonisation in the shortest period of 38 ± 0.5 days. However, only sawdust and bamboo dust supported the primodia formation. Complex medium supported mycelium growth in submerged cultures and 27.11 ± 0.43 g/L of mycelia was obtained after 2 weeks of cultivation at 28 °C and 200 rpm. Antioxidant potential in mushroom may be influenced by different cultivation and extraction methods. The different extracts from the wild and cultivated basidiocarps as well as mycelia were then tested for their antioxidant properties. Aqueous and ethanol extracts of mycelia and basidiocarps tested had varying levels of antioxidant activities. To conclude, domestication of wild G. neo-japonicum using agroresidues may ensure a continuous supply of G. neo-japonicum for its medicinal use while ensuring the conservation of this rare species.

  1. Morphogenesis of Pistillate Flowers of Cercidiphyllum japonicum (Cercidiphyllaceae)

    Institute of Scientific and Technical Information of China (English)

    Xiao-Ling Yan; Yi Ren; Xian-Hua Tian; Xiao-Hui Zhang

    2007-01-01

    Floral morphogenesis and the development of Cercidiphyllumjaponicum Sieb. et Zucc. were observed by scanning electron microscopy (SEM). The results showed that the pistillate inflorescences were congested spikes with the flowers arranged opposite. Great differences between the so-called "bract" and the vegetative leaf were observed both in morphogenesis and morphology. In morphogenesis, the "bract" primordium is crescent-shaped, truncated at the apex and not conduplicate,has no stipule primordium at the base but does have some inconspicuous teeth in the margin that are not glandular. The leaf primordium is triangular, cycloidal at the apex, conduplicate, has two stipule primordia at the base, has one gland-tooth at the apex occurring at first and some gland-teeth in the margin that occur later. In morphology, the "bract" is also different to the vegetative leaf in some characteristics that were also illustrated in the present paper. Based on the hypothesis that the bract is more similar to the vegetative leaf than the tepal, we considered that the so-called "bract" of C. japonicum might be the tepal of the pistillate flower in morphological nature. Therefore, each pistillate flower contains a tepal and a carpel.We did not find any trace of other floral organs in the morphogenesis of the pistillate flower. Therefore we considered that the unicarpellate status of extant Cercidiphyllum might be to highly reduce and advance characteristics that make the extant Cercidiphyllum isolated from both fossil Cercidiphyllum-like plants and its extant affinities.

  2. First Report of Leaf Rust Caused by Puccinia caricis in Farfugium japonicum in Korea.

    Science.gov (United States)

    Yun, Yeo Hong; Kwon, Hyuk Woo; Ahn, Hong Seok; Kim, Seong Hwan

    2015-09-01

    Farfugium japonicum is used in traditional medicine and as an edible herb in China and Korea. In July 2013, leaf spots were observed in F. japonicum seedlings at Ulleung Island, Gyeongsangbuk Province, Korea. Early symptoms on the leaf adaxial surface included roughly circular yellow spots that later developed brown, necrotic centers. The aecia were hypophyllous, cupulate, yellowish, 180~430 µm in diameter, clustered, and erumpent with a peridium with a recurved margin. The aeciospores were globoid, 14~17 × 13~16 µm, light yellow or colorless, and densely verrucose. The 28S rDNA sequence of the isolate was identical to each other and shared 99% identity with Puccinia caricis. This is the first report of rust caused by P. caricis in F. japonicum in Korea or elsewhere in the world.

  3. Chemical composition and antimicrobial activity of essential oil isolated from the cultured mycelia of Ganoderma japonicum

    Institute of Scientific and Technical Information of China (English)

    Dandan Liu; Zheng Hu; Zhigang Liu; Bo Yang; Wenjuan Tu; Liang Li

    2009-01-01

    Objective:To explore a new natural antibiotic. Methods:The chemical composition of the essential oil from Ganoderma japonicum (G. japonicum) mycelia was analyzed by gas chromatography-mass spectrometry(GC-MS). The antimicrobial activity of the oil was evaluated against eighteen microorganisms, including bacteria, mildew and yeast by using a disc diffusion method. Furthermore, the minimum inhibitory concentrations(MIC) and the minimum bactericidal concentrations(MBC) of the essential oil against twelve clinical pathogens were determined. Results:The main components of the oil were nerolidol, decadienal, linaiool and benzyl alcohol. The antimicrobial results indicated that the oil inhibited all the tested bacterium, especially Methicillin-resistant Staphylococcus aureus (MRSA) in which the antibacterial activity exhibited a MBC of 1.03 mg/ml. Conclusion: The essential oil of G.japonicum mycelium has significant inhibitory activity. It is a potential medicinal resource that can be used as a natural antibiotic.

  4. Complete genome sequence of the mitochondrial DNA of the river lamprey, Lethenteron japonicum.

    Science.gov (United States)

    Kawai, Yuri L; Yura, Kei; Shindo, Miyuki; Kusakabe, Rie; Hayashi, Keiko; Hata, Kenichiro; Nakabayashi, Kazuhiko; Okamura, Kohji

    2015-01-01

    Lampreys are eel-like jawless fishes evolutionarily positioned between invertebrates and vertebrates, and have been used as model organisms to explore vertebrate evolution. In this study we determined the complete genome sequence of the mitochondrial DNA of the Japanese river lamprey, Lethenteron japonicum, using next-generation sequencers. The sequence was 16,272 bp in length. The gene content and order were identical to those of the sea lamprey, Petromyzon marinus, which has been the reference among lamprey species. However, the sequence similarity was less than 90%, suggesting the need for the whole-genome sequencing of L. japonicum.

  5. Rock blocks

    OpenAIRE

    Turner, W.

    2007-01-01

    Consider representation theory associated to symmetric groups, or to Hecke algebras in type A, or to q-Schur algebras, or to finite general linear groups in non-describing characteristic. Rock blocks are certain combinatorially defined blocks appearing in such a representation theory, first observed by R. Rouquier. Rock blocks are much more symmetric than general blocks, and every block is derived equivalent to a Rock block. Motivated by a theorem of J. Chuang and R. Kessar in the case of sym...

  6. Synthesis of new 4-oxo-2-thioxo-1,2,3,4-tetrahydropyrimidine derivatives with an incorporated thiazolidinone moiety and testing their possible serine protease and cercarial elastase inhibitory effects with a possible prospective to block penetration of Schistosoma mansoni cercariae into the mice skin.

    Science.gov (United States)

    Bahgat, Mahmoud Mohamed; Maghraby, Amany Sayed; Heiba, Mogeda Emam; Ruppel, Andreas; Fathalla, Omar Abd-elfattah Mohamed

    2005-09-01

    5-Substituted 4-oxo-2-thioxo-1,2,3,4-tetrahydropyrimidine were synthesized by interaction of 4-oxo-2-thioxo-1,2,3,4-tetrahydropyrimidine-5-sulfonylhydrazide with some aldehydes to give the corresponding Schiff-bases, which after cyclization gave corresponding thiazolidinones. For some of the thiazolidinones, Mannich bases reaction was carried out. All the derivatives were tested for their possible inhibitory effect on Schistosoma mansoni cercarial elastase (CE). Only, N-(4-methylbenzyledine)-4-oxo-2-thioxo-1,2,3,4-tetrahydropyrimidine-5-sulfonylhydrazide was found to have potent inhibitory effect on the CE activity with IC50 = 264 microM. Upon its use as a paint for mice tails before infection with S. mansoni cercariae, the compound formulated in jojoba oil caused a significant reduction (93%; P-value = 0.0002) in the worm burden. IgG & IgM in mice sera were measured by using several S. mansoni antigens by ELISA. Sera from treated infected mice (TIM) 2, 4, and 6 weeks (W) post infection (PI) showed 1.2 folds lower, 1.2 folds higher, 1.7 folds lower IgM reactivity against soluble cercarial antigenic preparation (CAP), respectively, when compared with sera collected from infected untreated mice (IUM). Sera from TIM 2, 4, and 6WPI showed 1.3, 1.6, and 1.7 folds higher IgG reactivity, respectively against CAP than the IgG reactivity from IUM. Sera from TIM 2, 4 and 6WPI showed 1.5, 1.2 folds lower and 1.4 folds higher IgM reactivity, respectively against soluble worm antigenic preparation (SWAP) when compared with sera collected from IUM. Sera from TIM 2, 4, and 6WPI showed 1.4, 1 folds lower and 1 fold higher IgG reactivity, respectivley to SWAP when compared with sera from IUM. Sera from TIM 2, 4, and 6WPI had generaly lower IgM and IgG reactivities against soluble egg antigen (SEA) when compared with sera from IUM.

  7. Lethal and sublethal effects of thiamethoxam on the whitefly predator Serangium japonicum (Coleoptera: Coccinellidae) through different exposure routes.

    Science.gov (United States)

    Yao, Feng-Luan; Zheng, Yu; Zhao, Jian-Wei; Desneux, Nicolas; He, Yu-Xian; Weng, Qi-Yong

    2015-06-01

    Given expectations for a booming usage of thiamethoxam and increasing availability of the promising biological agent Serangium japonicum for the control of Bemisia tabaci in China, an evaluation of their compatibility is crucial for integrated pest management (IPM). This study examined the lethal and sublethal effects of thiamethoxam on S. japonicum through three exposure routes. An acute toxicity bioassay showed that LC50 values of thiamethoxam for S. japonicum through residue contact, egg-dip, and systemic treatment were 6.65, 4.37, and 2.43 mg AI L(-1), respectively. The prey consumption of S. japonicum given different densities of B. tabaci eggs under control, discontinuous, egg-dip and systemic exposure scenarios showed a good fit to a Type II functional response. Predation of S. japonicum was most affected under systemic exposure, followed by egg-dip, and discontinuous, which was only slightly affected. In all cases tested, however, predators recovered their predation capacity rapidly, either after 24h of exposure or 24h after the end of exposure. Thiamethoxam was highly toxic to S. japonicum regardless of exposure routes. Sublethal effects of thiamethoxam applied systemically or foliar both impaired the biological control of S. japonicum on B. tabaci. Therefore, thiamethoxam should be used with caution in IPM of B. tabaci.

  8. Construction of Hemoglobin Degradation Pathway of Schistosoma and Analysis of Conserved Domains of Pathway Key Enzymes%血吸虫血红蛋白降解途径的重构及关键酶蛋白保守结构域的分析

    Institute of Scientific and Technical Information of China (English)

    魏佳; 李园园; 李亦学; 于复东

    2011-01-01

    On the basis of hemoglobin degradation pathway of Plasmodium falciparum, hemoglobin degradation pathways of Schistosoma japonicum and Schistosoma mansoni were constructed using bioinformatic methods. Sequences and structures of conserved domains of pathway key enzymes were analyzed after pathway construction. The catalytic sites of predicted hemoglobinases of Schistosoma are highly conserved with Plasmodium in both sequence and structure level. This paper provides a theoretical foundation for Schistosoma hemoglobinase study, and analysis of their interactions with substrates or inhibitors in three-dimensional level.%利用生物信息学方法,以疟原虫的血红蛋白降解途径为基础,成功地构建出了日本血吸虫和曼氏血吸虫的血红蛋白降解途径,并对降解途径上的关键酶蛋白的保守结构域进行了序列及结构分析。预测得到的血吸虫血红蛋白降解酶保守结构域,与疟原虫相比,在序列上催化位点高度保守,在结构上空间相对位置较一致,为研究血吸虫血红蛋白降解酶,及其与底物或抑制剂在三维结构上的相互作用提供了理论基础。

  9. Isolation of genes (nif/hup cosmids) involved in hydrogenase and nitrogenase activities in Rhizobium japonicum.

    Science.gov (United States)

    Hom, S S; Graham, L A; Maier, R J

    1985-03-01

    Recombinant cosmids containing a Rhizobium japonicum gene involved in both hydrogenase (Hup) and nitrogenase (Nif) activities were isolated. An R. japonicum gene bank utilizing broad-host-range cosmid pLAFR1 was conjugated into Hup- Nif- R. japonicum strain SR139. Transconjugants containing the nif/hup cosmid were identified by their resistance to tetracycline (Tcr) and ability to grow chemoautotrophically (Aut+) with hydrogen. All Tcr Aut+ transconjugants possessed high levels of H2 uptake activity, as determined amperometrically. Moreover, all Hup+ transconjugants tested possessed the ability to reduce acetylene (Nif+) in soybean nodules. Cosmid DNAs from 19 Hup+ transconjugants were transferred to Escherichia coli by transformation. When the cosmids were restricted with EcoRI, 15 of the 19 cosmids had a restriction pattern with 13.2-, 4.0-, 3.0-, and 2.5-kilobase DNA fragments. Six E. coli transformants containing the nif/hup cosmids were conjugated with strain SR139. All strain SR139 transconjugants were Hup+ Nif+. Moreover, one nif/hup cosmid was transferred to 15 other R. japonicum Hup- mutants. Hup+ transconjugants of six of the Hup- mutants appeared at a frequency of 1.0, whereas the transconjugants of the other nine mutants remained Hup-. These results indicate that the nif/hup gene cosmids contain a gene involved in both nitrogenase and hydrogenase activities and at least one and perhaps other hup genes which are exclusively involved in H2 uptake activity.

  10. Expression of symbiotic genes of Rhizobium japonicum USDA 191 in other rhizobia.

    OpenAIRE

    Appelbaum, E R; McLoughlin, T J; O'Connell, M; Chartrain, N

    1985-01-01

    A 200-megadalton plasmid was mobilized from Rhizobium japonicum USDA 191 to other Rhizobium strains either that cannot nodulate soybeans or that form Fix- nodules on certain cultivars. The symbiotic properties of the transconjugants indicate that both soybean specificity for nodulation and cultivar specificity for nitrogen fixation are plasmid encoded.

  11. Characterization of a Functional Role of the Bradyrhizobium japonicum Isocitrate Lyase in Desiccation Tolerance

    Directory of Open Access Journals (Sweden)

    Jeong-Min Jeon

    2015-07-01

    Full Text Available Bradyrhizobium japonicum is a nitrogen-fixing symbiont of soybean. In previous studies, transcriptomic profiling of B. japonicum USDA110, grown under various environmental conditions, revealed the highly induced gene aceA, encoding isocitrate lyase (ICL. The ICL catalyzes the conversion of isocitrate to succinate and glyoxylate in the glyoxylate bypass of the TCA cycle. Here, we evaluated the functional role of B. japonicum ICL under desiccation-induced stress conditions. We purified AceA (molecular mass = 65 kDa from B. japonicum USDA110, using a His-tag and Ni-NTA column approach, and confirmed its ICL enzyme activity. The aceA mutant showed higher sensitivity to desiccation stress (27% relative humidity (RH, compared to the wild type. ICL activity of the wild type strain increased approximately 2.5-fold upon exposure to 27% RH for 24 h. The aceA mutant also showed an increased susceptibility to salt stress. Gene expression analysis of aceA using qRT-PCR revealed a 148-fold induction by desiccation, while other genes involved in the glyoxylate pathway were not differentially expressed in this condition. Transcriptome analyses revealed that stress-related genes, such as chaperones, were upregulated in the wild-type under desiccating conditions, even though fold induction was not dramatic (ca. 1.5–2.5-fold.

  12. Plant recognition of Bradyrhizobium japonicum nod factors. Final report, September 15, 1992--March 14, 1997

    Energy Technology Data Exchange (ETDEWEB)

    Stacey, G.

    1998-01-01

    This grant had three objectives: (1) isolate and identify the unique nod factor metabolites made by different wild-type B. japonicum strains; (2) investigate the biological activity of these unique nod factors, especially as it relates to host range; and (3) initiate studies to define the mechanism of plant recognition of the nod factors. This report summarizes the results of this research.

  13. Diagnostic performance of Schistosoma real-time PCR in urine samples from Kenyan children infected with Schistosoma haematobium

    DEFF Research Database (Denmark)

    Vinkeles Melchers, Natalie V. S.; van Dam, Govert J.; Shaproski, David

    2014-01-01

    BACKGROUND: In an effort to enhance accuracy of diagnosis of Schistosoma haematobium, this study explores day-to-day variability and diagnostic performance of real-time PCR for detection and quantification of Schistosoma DNA compared to other diagnostic tools in an endemic area before and after......, respectively. Based on the 'gold standard', PCR showed high sensitivity (>92%) as compared to >31% sensitivity for microscopy, both pre- and post-treatment. CONCLUSIONS/SIGNIFICANCE: Detection and quantification of Schistosoma DNA in urine by real-time PCR was shown to be a powerful and specific diagnostic...... tool for detection of S. haematobium infections, with less day-to-day variation and higher sensitivity compared to microscopy. The superior performance of PCR before, and two and 18 months post-treatment provides a compelling argument for PCR as an accurate and reproducible tool for monitoring...

  14. CD8α¯ DC is the major DC subset which mediates inhibition of allergic responses by Schistosoma infection.

    Science.gov (United States)

    Liu, J-Y; Lu, P; Hu, L-Z; Shen, Y-J; Zhu, Y-J; Ren, J-L; Ji, W-H; Zhang, X-Z; Wu, Z-Q; Yang, X-Z; Yang, J; Li, L-Y; Yang, X; Liu, P-M

    2014-12-01

    Our and others' previous studies have shown that Schistosoma japonicum (SJ) infection can inhibit allergic reactions. We recently reported that DCs played an important role in SJ infection-mediated inhibition of allergy, which was associated with enhanced IL-10 and T regulatory cell responses. Here, we further compared the role of CD8α(+) DC and CD8α(-) DC subsets for the inhibitory effect. We sorted CD8α(+) DC (SJCD8α(+) DC) and CD8α(-) DC (SJCD8α(-) DC) from SJ-infected mice and tested their ability to modulate allergic responses in vivo. The data showed that the adoptive transfer of SJCD8α(-) DC was much more efficient than SJCD8α(+) DC for the suppression of allergic airway eosinophilia, mucus overproduction, antigen-specific IgE responses, and Th2 cytokines (IL-4 and IL-5). More importantly, we found that the transfer of SJCD8α(-) DC, but not SJCD8α(+) DC, significantly increased IL-10 and TGF-β production following OVA exposure. As control, the transfer of DC subsets from naïve mice had no significant effect on allergic inflammation. In addition, SJCD8α-DC expressed significantly higher IL-10 but lower IL-12, CD80 and CD86 than SJCD8α(+) DC, fitting a tolerogenic phenotype. The results suggest that CD8α(-) DC is the predominant DC subset which is involved in the parasitic infection-mediated inhibition of allergic inflammation and possibly through enhancing immunomodulatory cytokine (IL-10 and TGF-β) production.

  15. Characterization of SR3 reveals abundance of non-LTR retrotransposons of the RTE clade in the genome of the human blood fluke, Schistosoma mansoni

    Directory of Open Access Journals (Sweden)

    Brindley Paul J

    2005-11-01

    Full Text Available Abstract Background It is becoming apparent that perhaps as much as half of the genome of the human blood fluke Schistosoma mansoni is constituted of mobile genetic element-related sequences. Non-long terminal repeat (LTR retrotransposons, related to the LINE elements of mammals, comprise much of this repetitive component of the schistosome genome. Of more than 12 recognized clades of non-LTR retrotransposons, only members of the CR1, RTE, and R2 clades have been reported from the schistosome genome. Results Inspection of the nucleotide sequence of bacterial artificial chromosome number 49_J_14 from chromosome 1 of the genome of Schistosoma mansoni (GenBank AC093105 revealed the likely presence of several RTE-like retrotransposons. Among these, a new non-LTR retrotransposon designated SR3 was identified and is characterized here. Analysis of gene structure and phylogenetic analysis of both the reverse transcriptase and endonuclease domains of the mobile element indicated that SR3 represented a new family of RTE-like non-LTR retrotransposons. Remarkably, two full-length copies of SR3-like elements were present in BAC 49-J-14, and one of 3,211 bp in length appeared to be intact, indicating SR3 to be an active non-LTR retrotransposon. Both were flanked by target site duplications of 10–12 bp. Southern hybridization and bioinformatics analyses indicated the presence of numerous copies (probably >1,000 of SR3 interspersed throughout the genome of S. mansoni. Bioinformatics analyses also revealed SR3 to be transcribed in both larval and adult developmental stages of S. mansoni and to be also present in the genomes of the other major schistosome parasites of humans, Schistosoma haematobium and S. japonicum. Conclusion Numerous copies of SR3, a novel non-LTR retrotransposon of the RTE clade are present in the genome of S. mansoni. Non-LTR retrotransposons of the RTE clade including SR3 appear to have been remarkably successful in colonizing, and

  16. De novo transcriptome assembly of a fern, Lygodium japonicum, and a web resource database, Ljtrans DB.

    Science.gov (United States)

    Aya, Koichiro; Kobayashi, Masaaki; Tanaka, Junmu; Ohyanagi, Hajime; Suzuki, Takayuki; Yano, Kenji; Takano, Tomoyuki; Yano, Kentaro; Matsuoka, Makoto

    2015-01-01

    During plant evolution, ferns originally evolved as a major vascular plant with a distinctive life cycle in which the haploid and diploid generations are completely separated. However, the low level of genetic resources has limited studies of their physiological events, as well as hindering research on the evolutionary history of land plants. In this study, to identify a comprehensive catalog of transcripts and characterize their expression traits in the fern Lygodium japonicum, nine different RNA samples isolated from prothalli, trophophylls, rhizomes and sporophylls were sequenced using Roche 454 GS-FLX and Illumina HiSeq sequencers. The hybrid assembly of the high-quality 454 GS-FLX and Illumina HiSeq reads generated a set of 37,830 isoforms with an average length of 1,444 bp. Using four open reading frame (ORF) predictors, 38,142 representative ORFs were identified from a total of 37,830 transcript isoforms and 95 contigs, which were annotated by searching against several public databases. Furthermore, an orthoMCL analysis using the protein sequences of L. japonicum and five model plants revealed various sets of lineage-specific genes, including those detected among land plant lineages and those detected in only L. japonicum. We have also examined the expression patterns of all contigs/isoforms, along with the life cycle of L. japonicum, and identified the tissue-specific transcripts using statistical expression analyses. Finally, we developed a public web resource, the L. japonicum transcriptome database at http://bioinf.mind.meiji.ac.jp/kanikusa/, which provides important opportunities to accelerate molecular research in ferns.

  17. Structural studies of Schistosoma mansoni adenylate kinases

    Energy Technology Data Exchange (ETDEWEB)

    Marques, I.A. [Universidade Federal de Goias (UFG), Goiania, GO (Brazil); Pereira, H.M.; Garrat, R.C. [Universidade de Sao Paulo (USP-SC), Sao Carlos, SP (Brazil)

    2012-07-01

    Full text: Parasitic diseases are a major cause of death in developing countries, however receive little or no attention from pharmaceutical companies for the development of novel therapies. In this respect, the Center for Structural Molecular Biology (CBME) of the Institute of Physics of Sao Carlos (IFSC / USP) has developed expertise in all stages of the development of active compounds against target enzymes from parasitic diseases. The present work focuses on the adenylate kinase enzymes (ADK's) from Schistosoma mansoni. These enzymes are widely distributed and catalyze the reaction of phosphoryl exchange between nucleotides in the reaction 2ADP to ATP + AMP, which is critical for the cells life cycle. Due to the particular property of the reaction catalyzed, the ADK's are recognized as reporters of the cells energetic state, translating small changes in the balance between ATP and ADP into a large change in concentration of AMP. The genome of S. mansoni was recently sequenced by the Sanger Center in England. On performing searches for genes encoding adenylate kinases we found two such genes. The corresponding gene products were named ADK1 (197 residues) and ADK2 (239 residues), and the two sequences share only 28 percent identity. Both have been cloned into the pET-28a(+)vector, expressed in E. coli and purified. Preliminary tests of activity have been performed only for ADK1 showing it to be catalytically active. Crystallization trials were performed for both proteins and thus far, crystals of ADK1 have been obtained which diffract to 2.05 at the LNLS beamline MX2 and the structure solved by molecular replacement. Understanding, at the atomic level, the function of these enzymes may help in the development of specific inhibitors and may provide tools for developing diagnostic tests for schistosomiasis. (author)

  18. Influence of oxygen on DNA binding, positive control, and stability of the Bradyrhizobium japonicum NifA regulatory protein.

    OpenAIRE

    1991-01-01

    Central to the genetic regulatory circuit that controls Bradyrhizobium japonicum nif and fix gene expression is the NifA protein. NifA activates transcription of several nif and fix genes and autoregulates its expression during symbiosis in soybean root nodules or in free-living microaerobic conditions. High O2 tensions result in the lack of nif expression, possibly by inactivation of NifA through oxidation of an essential metal cofactor. Several B. japonicum nif and fix promoters have upstre...

  19. Population Blocks.

    Science.gov (United States)

    Smith, Martin H.

    1992-01-01

    Describes an educational game called "Population Blocks" that is designed to illustrate the concept of exponential growth of the human population and some potential effects of overpopulation. The game material consists of wooden blocks; 18 blocks are painted green (representing land), 7 are painted blue (representing water); and the remaining…

  20. Lack of glyphosate resistance gene transfer from Roundup Ready soybean to Bradyrhizobium japonicum under field and laboratory conditions.

    Science.gov (United States)

    Isaza, Laura Arango; Opelt, Katja; Wagner, Tobias; Mattes, Elke; Bieber, Evi; Hatley, Elwood O; Roth, Greg; Sanjuán, Juan; Fischer, Hans-Martin; Sandermann, Heinrich; Hartmann, Anton; Ernst, Dieter

    2011-01-01

    A field study was conducted at the Russell E. Larson Agricultural Research Center to determine the effect of transgenic glyphosate-resistant soybean in combination with herbicide (Roundup) application on its endosymbiont Bradyrhizobium japonicum. DNA of bacteroids from isolated nodules was analysed for the presence of the transgenic 5-enolpyruvylshikimate-3-phosphate synthase (CP4-EPSPS) DNA sequence using polymerase chain reaction (PCR). To further assess the likelihood that the EPSPS gene may be transferred from the Roundup Ready (RR) soybean to B. japonicum, we have examined the natural transformation efficiency of B. japonicum strain 110spc4. Analyses of nodules showed the presence of the transgenic EPSPS DNA sequence. In bacteroids that were isolated from nodules of transgenic soybean plants and then cultivated in the presence of glyphosate this sequence could not be detected. This indicates that no stable horizontal gene transfer (HGT) of the EPSPS gene had occurred under field conditions. Under laboratory conditions, no natural transformation was detected in B. japonicum strain 110spc4 in the presence of various amounts of recombinant plasmid DNA. Our results indicate that no natural competence state exists in B. japonicum 110spc4. Results from field and laboratory studies indicate the lack of functional transfer of the CP4-EPSPS gene from glyphosate-tolerant soybean treated with glyphosate to root-associated B. japonicum.

  1. Assessment of Trichogramma japonicum and T. chilonis as Potential Biological Control Agents of Yellow Stem Borer in Rice.

    Science.gov (United States)

    Tang, Rui; Babendreier, Dirk; Zhang, Feng; Kang, Min; Song, Kai; Hou, Mao-Lin

    2017-02-08

    Two species of Trichogramma wasps were assessed for their effectiveness against yellow stem borer Scirpophaga incertulas. A laboratory cage test with T. japonicum and T. chilonis showed that both species parasitized yellow stem borer egg masses at 60.0% ± 9.13% and 40.7% ± 7.11%, respectively, with egg parasitism rates of 15.8% ± 22.2% for T. japonicum and 2.8% ± 5.0% for T. chilonis. Once the host eggs were parasitized, emergence rates were high for both species (95.7% ± 0.12% for T. japonicum and 100% for T. chilonis). In paddy field trials, the two Trichogramma species were released at three densities (50,000/ha, 100,000/ha and 200,000/ha) in Southwestern China. Egg mass parasitism was 9% ± 7.7% for T. japonicum and 15% ± 14.1% for T. chilonis, and again only a relatively small fraction of eggs was successfully parasitized. No clear conclusion could be drawn on the most efficient release rate as no significant differences were found among the three release rates. A comparison of field-collected T. japonicum with T. japonicum and T. chilonis mass reared on Corcyra cephalonica showed significantly larger body size and ovipositor length in field-collected wasps, suggesting potentially higher effectiveness on yellow stem borer eggs after at least one generation on the target host. Factors contributing to the low field parasitism rates are discussed.

  2. Assessment of Trichogramma japonicum and T. chilonis as Potential Biological Control Agents of Yellow Stem Borer in Rice

    Directory of Open Access Journals (Sweden)

    Rui Tang

    2017-02-01

    Full Text Available Two species of Trichogramma wasps were assessed for their effectiveness against yellow stem borer Scirpophaga incertulas. A laboratory cage test with T. japonicum and T. chilonis showed that both species parasitized yellow stem borer egg masses at 60.0% ± 9.13% and 40.7% ± 7.11%, respectively, with egg parasitism rates of 15.8% ± 22.2% for T. japonicum and 2.8% ± 5.0% for T. chilonis. Once the host eggs were parasitized, emergence rates were high for both species (95.7% ± 0.12% for T. japonicum and 100% for T. chilonis. In paddy field trials, the two Trichogramma species were released at three densities (50,000/ha, 100,000/ha and 200,000/ha in Southwestern China. Egg mass parasitism was 9% ± 7.7% for T. japonicum and 15% ± 14.1% for T. chilonis, and again only a relatively small fraction of eggs was successfully parasitized. No clear conclusion could be drawn on the most efficient release rate as no significant differences were found among the three release rates. A comparison of field-collected T. japonicum with T. japonicum and T. chilonis mass reared on Corcyra cephalonica showed significantly larger body size and ovipositor length in field-collected wasps, suggesting potentially higher effectiveness on yellow stem borer eggs after at least one generation on the target host. Factors contributing to the low field parasitism rates are discussed.

  3. Bioactivity of miltefosine against aquatic stages of Schistosoma mansoni, Schistosoma haematobium and their snail hosts, supported by scanning electron microscopy

    Directory of Open Access Journals (Sweden)

    El Bardicy Samia

    2011-05-01

    Full Text Available Abstract Background Miltefosine, which is the first oral drug licensed for the treatment of leishmaniasis, was recently reported to be a promising lead compound for the synthesis of novel antischistosomal derivatives with potent activity in vivo against different developmental stages of Schistosoma mansoni. In this paper an in vitro study was carried out to investigate whether it has a biocidal activity against the aquatic stages of Schistosoma mansoni and its snail intermediate host, Biomphalaria alexandrina , thus being also a molluscicide. Additionally, to see whether miltefosine can have a broad spectrum antischistosomal activity, a similar in vitro study was carried out on the adult stage of Schistosoma haematobium, the second major human species, its larval stages and snail intermediate host, Bulinus truncutes. This was checked by scanning electron microscopy. Results Miltefosine proved to have in vitro ovicidal, schistolarvicidal and lethal activity on adult worms of both Schistosoma species and has considerable molluscicidal activity on their snail hosts. Scanning electron microscopy revealed several morphological changes on the different stages of the parasite and on the soft body of the snail, which further strengthens the current evidence of miltefosine's activity. This is the first report of mollusicidal activity of miltefosine and its in vitro schistosomicidal activity against S.haematobium. Conclusions This study highlights miltefosine not only as a potential promising lead compound for the synthesis of novel broad spectrum schistosomicidal derivatives, but also for molluscicidals.

  4. 南京所见一种裂体吸虫与日本血吸虫形态及生活史比较%DESCRIPTION OF A KIND OF SCHISTOSOMA IN NANJING REGION

    Institute of Scientific and Technical Information of China (English)

    徐国余; 陈天明; 赵勇进; 陈广梅; 庞浩

    2001-01-01

    In the paper, some morphological and biological materials of a kind of Schistosoma obtained from mice or rabbits experimentally infected with cercariae from natural snail living in Nanjing are presented as follows: (1) Adult worms superficially resemble to Schistosoma japonicum but smaller in size. For instance, male worms from mice 45d after infection measure (7.14±0.64) mm × (0.58±0.03) mm. In average, oral sucker is 0.54 mm ×0.59 mm, and ventral sucker 0.65 mm×0.68 mm which goes beyond the body width and no pronounced rim found. There are usually 8 testes in two series. Dosal integument presents densely clublike spines. Females from mice 45d after infection measure (7.19±1.12) mm×(0.37±0.02) mm. Oral suckers are bigger than ventral ones. The uterus contains as many as 20~80 eggs at one time. Egg has lateral hook-ring spine. (2) The typical habitat for oviposition is in small and terminal veins of superior and inferior mesenteric veins, potal veins in liver and vesical venous plexus. (3) In mice, the patent period is 32.6 d±1.2 d which is shorter than that of S. japonicum while in Oncomelania snails it is 9 wks under 26℃ which is 1 wk longer tahn that of S. japonicum. (4) Study on snail compatibility showed that infection rate of this Schistosma was 3.7%~6.6% but of S. japonicum 31.7%~46.6%.%用自然环境中钉螺逸出的尾蚴感染小鼠和兔,获得了一种比日本血吸虫小的裂体吸虫。雄虫大小(7.14±0.64)mm×(0.58±0.03)mm,口吸盘较大0.54mm×0.59mm;腹吸盘更大0.65mm×0.68mm,超过虫体宽度,且圆边构造不明显;睾丸4~8个,多为8个,重叠排成两行;背部有密集杵状小棘。雌虫(7.19±1.12)mm×(0.37±0.02)mm,子宫内有虫卵20~80个。雌雄合抱寄生在肠系膜下静脉、肠系膜上静脉、肝内门脉血管和膀胱静脉丛的小静脉和末梢静脉中。虫卵有钩状或环状微侧刺。终宿主小鼠的虫卵开放前期32.6d±1.2d,比日本血吸虫短1.4d

  5. NCBI nr-aa BLAST: CBRC-MMUR-01-0199 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUR-01-0199 emb|CAX75788.1| Tubulin beta-2C chain [Schistosoma japonicum] emb...|CAX75790.1| Tubulin beta-2C chain [Schistosoma japonicum] emb|CAX75791.1| Tubulin beta-2C chain [Schistosoma japonicum] CAX75788.1 2e-31 82% ...

  6. Allelopathy of the invasive plant Bidens frondosa on the seed germination of Geum japonicum var. chinense.

    Science.gov (United States)

    Wang, X F; Hassani, D; Cheng, Z W; Wang, C Y; Wu, J

    2014-12-12

    Five gradient concentrations (0.02, 0.04, 0.06, 0.08, and 0.10 g/mL) of leaching liquors from the roots, stems, and leaves of the invasive plant Bidens frondosa were used as conditioning fluid to examine its influence on seed germination conditions of the native plant Geum japonicum var. chinense in Huangshan. All leaching liquors of organs suppressed the seed germination of Geum japonicum var. chinense and reduced the final germination percentage and rate, and increased the germination inhibition rate, with a bimodal dependence on concentration. The leaching liquor inhibited the seed germination significantly at the concentration of 0.02 g/mL respectively. The seed germination was also inhibited as the concentration reached to 0.04 g/mL and beyond. Hence the allelopathic effects of the organs were significantly enhanced respectively. This phenomenon represented the presence of allelopathy substances in the root, stem and leaf of Bidens frondosa.

  7. Zitongdong Block

    Institute of Scientific and Technical Information of China (English)

    1996-01-01

    @@ The Zitongdong Block (Eastern Zitong Block) is located in the northwest of the Sichuan Basin. Tectonically, it is situated in the east part of Zitong Depression, southeast of mid-Longmenshan folded and faulted belt( as shown on Fig. 8 ), covering an area of 1 730 km2. The traffic is very convenient, the No. 108 national highway passes through the north of the block. Topographically, the area belongs to low hilly land at the elevation of 500-700 m.

  8. Transmissible Resistance to Penicillin G, Neomycin, and Chloramphenicol in Rhizobium japonicum1

    Science.gov (United States)

    Cole, Michael A.; Elkan, Gerald H.

    1973-01-01

    The genetic basis for resistance to a number of antibiotics was examined in Rhizobium japonicum. Resistance to penicillin G, neomycin, and chloramphenicol appears to be mediated by an extrachromosomal element similar to that found in the Enterobacteriaceae. Resistance to these antibiotics was eliminated from cells by treatment with acridine orange, and resistance to all three antibiotics could be transferred en bloc to Agrobacterium tumefaciens under conditions excluding transformation or transduction as possible genetic mechanisms. PMID:4491197

  9. Dynamic observation of splenocyte apoptosis in mice immunized with recombinant vaccine Bifidobacterium bifidum pGEX-Sj14-3-3 of Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    张宁

    2013-01-01

    Objective To investigate the effects of recombinant vaccine Bifidobacterium bifidum(Bb) pGEX-Sj14-3-3 on splenocyte apoptosis in BALB/c mice. Methods Ninety-six BALB/c mice were randomly divided into two groups according to their body mass: per os group(PO) and

  10. Functional diversity of Cercidiphyllum japonicum, communities in the Shennongjia Reserve, central China

    Institute of Scientific and Technical Information of China (English)

    Jintun Zhang; Bin Zhang; Zhiying Qian

    2015-01-01

    Functional diversity is significant to ecological processes of plant communities. We analyzed the variation of functional diversity of endangered species, Cercidi-phyllum japonicum, communities along an elevational gradient in the Shennongjia Reserve, central China. Sixty plots of 10 × 20 m from 1,350 to 2,050 m were set up and species composition, traits and environmental variables were measured and recorded. These data were analyzed using five functional diversity indices, functional attribute diversity, modified functional attribute diversity, plot based functional diversity, community based functional diversity and Rao’s functional diversity indices (Rao’s index), Functional diversities of C. japonicum communities were rich and varied greatly. Functional diversity declined non-linearly with increasing elevation. Functional diversity was significantly correlated with species richness and hetero-geneity. Elevation was a key environmental variable influencing functional diversity and species diversity. The five functional diversity indices were all effective for measuring functional diversity of communities. Functional diversity can be used as an indicator of conservation effi-ciency of endangered species such as C. japonicum.

  11. Indirect Effects of Oral Tolerance Inhibit Pulmonary Granulomas to Schistosoma mansoni Eggs

    Directory of Open Access Journals (Sweden)

    Geraldo Magela Azevedo

    2012-01-01

    Full Text Available Parenteral injection of tolerated proteins into orally tolerant mice inhibits the initiation of immunological responses to unrelated proteins and blocks severe chronic inflammatory reactions of immunological origin, such as autoimmune reactions. This inhibitory effect which we have called “indirect effects of oral tolerance” is also known as “bystander suppression.” Herein, we show that i.p. injection of OVA + Al(OH3 minutes before i.v. injection of Schistosoma mansoni eggs into OVA tolerant mice blocked the increase of pulmonary granulomas. In addition, the expression of ICAM-1 in lung parenchyma in areas outside the granulomas of OVA-orally tolerant mice was significantly reduced. However, at day 18 after granuloma induction there was no difference in immunofluorescency intensity to CD3, CD4, F4/80, andα-SMA per granuloma area of tolerant and control groups. Reduction of granulomas by reexposure to orally tolerated proteins was not correlated with a shift in Th-1/Th-2 cytokines in serum or lung tissue extract.

  12. Inheritance of Schistosoma mansoni infection incompatibility in Biomphalaria alexandrina snails

    OpenAIRE

    Iman F Abou El Naga; Eissa, Maha M.; Shereen F Mossallam; Safaa I Abd El-Halim

    2010-01-01

    In this study, we looked at the inheritance of susceptibility and resistance to Schistosoma mansoni infection in the first generation of crossbred Biomphalaria alexandrina snails. Our ultimate goal is to use such information to develop a biological method of controlling schistosomiasis. We infected laboratory-bred snails with S. mansoni miracidia and examined cercarial shedding to determine susceptibility and resistance. Five parental groups were used: Group I contained 30 susceptible snails,...

  13. Recombinant protein of Schistosoma japonicum calpain induced antibody production and detection of schistosoma infection%重组Calpain蛋白诱导抗体产生及检测日本血吸虫感染的研究

    Institute of Scientific and Technical Information of China (English)

    张仁利; 吴少庭; 高世同; 黄达娜; 袁仕善

    2003-01-01

    目的研究在大肠杆菌中表达的日本血吸虫Calpain蛋白的免疫原性及其在日本血吸虫病诊断上的应用. 方法用重组Calpain蛋白免疫BALB/c小鼠,ELISA法测定特异性IgG抗体的动态变化及其IgG亚型(IgG1,IgG2a,IgG2b,IgG3)产生特征;同时用重组Calpain蛋白作为诊断抗原,粪检血吸虫病阳性患者血清作为一抗,肝吸虫阳性患者血清作为考核交叉反应血清. 结果重组Calpain抗原免疫小鼠后,产生了一个极高的抗Calpain特异性抗体,免疫4周IgG类抗体达到高峰,与对照组鼠相比较免疫鼠血清中IgG1,IgG2a,IgG2b特异性抗体也显著上升;Calpain重组抗原血清检测日本血吸虫和粪检的符合率为100%,粪检阳性EPG低的个体抗Calpain抗体滴度高,EPG高的患者抗Calpain抗体滴度反而低,与肝吸虫的交叉反应率为37%. 结论日本血吸虫Calpain是一个具有免疫原性的蛋白,能够激发宿主产生高水平的免疫球蛋白,能够敏感地测定日本血吸虫的感染,提示Calpain可以发展成为日本血吸虫病的诊断抗原和日本血吸虫疫苗候选分子.

  14. 曼氏和日本血吸虫副肌球蛋白的抗原肽研究%Studies on Antigenic Peptides of Paramyosins of Schistosoma mansoni and Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    杨俊海; 许家喜

    2002-01-01

    副肌球蛋白是世界卫生组织选取的用来发展血吸虫疫苗的重要候选蛋白之一.在计算机辅助下,应用PCGene程序,根据曼氏和日本血吸虫副肌球蛋白的氨基酸序列,通过对其亲水性、柔韧性、可接近性和二级结构分析预测出8个抗原肽,并用固相法进行了合成.经Dot-ELISA法测定,其中的2个对抗日本血吸虫免疫球蛋白多克隆抗体(抗-Sj-IgG-PcAb)显示出抗原性,可作为抗血吸虫病合成多肽疫苗的候选肽段.

  15. Functional characterization of the Bradyrhizobium japonicum modA and modB genes involved in molybdenum transport.

    Science.gov (United States)

    Delgado, María J; Tresierra-Ayala, Alvaro; Talbi, Chouhra; Bedmar, Eulogio J

    2006-01-01

    A modABC gene cluster that encodes an ABC-type, high-affinity molybdate transporter from Bradyrhizobium japonicum has been isolated and characterized. B. japonicum modA and modB mutant strains were unable to grow aerobically or anaerobically with nitrate as nitrogen source or as respiratory substrate, respectively, and lacked nitrate reductase activity. The nitrogen-fixing ability of the mod mutants in symbiotic association with soybean plants grown in a Mo-deficient mineral solution was severely impaired. Addition of molybdate to the bacterial growth medium or to the plant mineral solution fully restored the wild-type phenotype. Because the amount of molybdate required for suppression of the mutant phenotype either under free-living or under symbiotic conditions was dependent on sulphate concentration, it is likely that a sulphate transporter is also involved in Mo uptake in B. japonicum. The promoter region of the modABC genes has been characterized by primer extension. Reverse transcription and expression of a transcriptional fusion, P(modA)-lacZ, was detected only in a B. japonicum modA mutant grown in a medium without molybdate supplementation. These findings indicate that transcription of the B. japonicum modABC genes is repressed by molybdate.

  16. CO-INOCULAÇÃO E MODOS DE APLICAÇÃO DE Bradyrhizobium japonicum e Azospirillum brasilense E ADUBAÇÃO NITROGENADA NA NODULAÇÃO DAS PLANTAS E RENDIMENTO DA CULTURA DA SOJA

    Directory of Open Access Journals (Sweden)

    Alessandro Lucca Braccini

    2016-03-01

    Full Text Available As new cultivation techniques arise, crops require the development of new technologies and management practices that aim higher yields with lower production cost. Thus, the objective of this study was to evaluate the efficiency of co-inoculation and different ways of application of Bradyrhizobium japonicum and Azospirillum brasilense and nitrogen fertilization on nodulation and yield of soybean. The experiment was established in November 2013 in an area located in the Iguatemi Experimental Farm, of State University of Maringá (UEM. The design was a randomized block with four replications. The treatments consisted of a control, fertilization with nitrogen (200 kg N ha-1, standard inoculation in seeds (liquid and peat inoculant, co-inoculation on seed and co-inoculation at sowing, using different doses of inoculant B. japonicum and A. brasilense. The following determinations were carried out in the field and in the laboratory: number of nodes at flowering, mass of nodules at flowering, the shoot dry mass, nitrogen content in the shoot, grain nitrogen content, number of pods/plant, thousand grain mass and yield. The results showed that inoculation through seed treatment with liquid inoculant presents an adequate option to farmers, in the conditions of this experiment. Nitrogen fertilization did not increase yield and yield components, being unnecessary.

  17. Zitongxi Block

    Institute of Scientific and Technical Information of China (English)

    1996-01-01

    @@ Zitongxi Block (Western Zitong Block), is located in Zitong County, northwest of Sichuan Province (as shown on Fig. 8 ). Geologically. it is situated in the Zitong Depression, southwest of the middle Longmenshan faulted and folded belt, covering an area of 1 830 km2. Transportation is very convenient. A crisscross network of highways run through the block and the Baocheng railway is nearby. The climate is moderate. Most area belongs to hilly land with the elevation of 500-600 m.The Tongjiang River runs across the area.

  18. Homologous modeling and function analysis on thioredoxin glutathione reductase from Schistosoma j aponicum%日本血吸虫硫氧还蛋白谷胱甘肽还原酶同源建模及功能分析

    Institute of Scientific and Technical Information of China (English)

    黄经纬; 郑予桐; 李家璜; 华子春

    2014-01-01

    目的:对日本血吸虫硫氧还蛋白谷胱甘肽还原酶(TGR)进行结构及功能分析。方法基于结构序列比较利用Swiss-Pdbviewer构建了日本血吸虫的TGR同源结构模型,并对模型进行结构评估;分析日本血吸虫TGR与底物结合时可能的位点,比较这些位点在不同来源TGR中的异同。结果日本血吸虫 TGR结构在 PROCHECK评估中被证实可靠;位点比较分析表明NADPH、GDS结合区是保守的位点;GSH结合区存在特异性。结论作用于GDS、NADPH结合区的其它来源的TGR抑制剂可能对日本血吸虫TGR也有作用;GSH结合区是设计寄生虫TGR特异性抑制剂的潜在靶点之一;TGR的C末端对电子传递起着重要作用并可能参与底物的结合,因而阻断日本血吸虫TGR的C末端摆动的抑制剂将可能有效地抑制日本血吸虫TGR活性。%To explore the structure and function of thioredoxin glutathione reductase (TGR) from Schistosoma j aponi-cum ,the homologous model of TGR in Schistosoma j aponicum was constructed by Swiss-Pdbviewer based on sequence and structure alignment .The potential substrates binding sites of TGR were analyzed and these sites of various TGRs were also as-sessed .Our results showed that the homologous model of Schistosoma japonicum TGR based on Schistosoma mansoni TGR structure was proved to be reasonable by PROCHECK program .Analysis of binding sites showed that NADPH and GDS bind-ing sites were conservative sites and GSH binding site was a specific site for parasite .Our data suggested that inhibitors which work in NADPH and GDS binding sites of other various TGRs may also interact with TGR form Schistosoma j aponicum .GSH binding region might be one of the potential targets for design of specific inhibitors of parasite TGRs .In addition ,C-terminal of TGR plays an important role in electron transfer and may participate in the binding of the substrate .Thus compound inhibiting swing of C-terminal could

  19. Chengzikou Block

    Institute of Scientific and Technical Information of China (English)

    1996-01-01

    @@ Chengzikou Block is located in the north of Hekou district, Dongying City, Shandong Province, adjacent to Bohai Bay. It can be geographically divided into three units: onshore, transitional zone and offshore ultrashallow zone, totally covering an area of 470 km2. The southern onshore area is low and gentle in topography;the northern shallow sea is at water depths of 2-8 m below sea level, and the transitional zone occupies more than 60% of the whole block. The climate belongs to temperate zone with seasonal wind. Highways are welldeveloped here, and the traffic is very convenient. The Chengzikou Block is about 80 km away from Dongying City and 290 km from Jinan City in the south. The northern offshore area of the block is 160 km away from Longkou port in the east and only 38 km away in the west from Zhuangxi port.

  20. Longmenshan Block

    Institute of Scientific and Technical Information of China (English)

    1996-01-01

    @@ Longmenshan Block is located in Jiange County of Jiangyou City in the northwest of Sichuan Basin. covering an area of 2 628 km2. Geologically, it is situated in the Mid-Longmenshan fault and fold belt, neighbouring Zitong Depression in its southeast. There are mountains surrounding its northwest , the rest area being hilly land,with the elevation of 500-700 m. The BaoCheng railway and the No. 108 highway run through the block, the traffic is very convenient.

  1. Decline in infection-related morbidities following drug-mediated reductions in the intensity of Schistosoma infection: A systematic review and meta-analysis

    Science.gov (United States)

    Andrade, Gisele; Bertsch, David J.; Gazzinelli, Andrea

    2017-01-01

    Background Since 1984, WHO has endorsed drug treatment to reduce Schistosoma infection and its consequent morbidity. Cross-sectional studies suggest pre-treatment correlation between infection intensity and risk for Schistosoma-related pathology. However, evidence also suggests that post-treatment reduction in intensity may not reverse morbidity because some morbidities occur at all levels of infection, and some reflect permanent tissue damage. The aim of this project was to systematically review evidence on drug-based control of schistosomiasis and to develop a quantitative estimate of the impact of post-treatment reductions in infection intensity on prevalence of infection-associated morbidity. Methodology/Principal findings This review was registered at inception with PROSPERO (CRD42015026080). Studies that evaluated morbidity before and after treatment were identified by online searches and searches of private archives. Post-treatment odds ratios or standardized mean differences were calculated for each outcome, and these were correlated to treatment-related egg count reduction ratios (ERRs) by meta-regression. A greater ERR correlated with greater reduction in odds of most morbidities. Random effects meta-analysis was used to derive summary estimates: after treatment of S. mansoni and S. japonicum, left-sided hepatomegaly was reduced by 54%, right-sided hepatomegaly by 47%, splenomegaly by 37%, periportal fibrosis by 52%, diarrhea by 53%, and blood in stools by 75%. For S. haematobium, hematuria was reduced by 92%, proteinuria by 90%, bladder lesions by 86%, and upper urinary tract lesions by 72%. There were no consistent changes in portal dilation or hemoglobin levels. In sub-group analysis, age, infection status, region, parasite species, and interval to follow-up were associated with meaningful differences in outcome. Conclusion/Significance While there are challenges to implementing therapy for schistosomiasis, and praziquantel therapy is not fully

  2. Survival and Competitiveness of Bradyrhizobium japonicum Strains 20 Years after Introduction into Field Locations in Poland.

    Science.gov (United States)

    Narożna, Dorota; Pudełko, Krzysztof; Króliczak, Joanna; Golińska, Barbara; Sugawara, Masayuki; Mądrzak, Cezary J; Sadowsky, Michael J

    2015-08-15

    It was previously demonstrated that there are no indigenous strains of Bradyrhizobium japonicum forming nitrogen-fixing root nodule symbioses with soybean plants in arable field soils in Poland. However, bacteria currently classified within this species are present (together with Bradyrhizobium canariense) as indigenous populations of strains specific for nodulation of legumes in the Genisteae tribe. These rhizobia, infecting legumes such as lupins, are well established in Polish soils. The studies described here were based on soybean nodulation field experiments, established at the Poznań University of Life Sciences Experiment Station in Gorzyń, Poland, and initiated in the spring of 1994. Long-term research was then conducted in order to study the relation between B. japonicum USDA 110 and USDA 123, introduced together into the same location, where no soybean rhizobia were earlier detected, and nodulation and competitive success were followed over time. Here we report the extra-long-term saprophytic survival of B. japonicum strains nodulating soybeans that were introduced as inoculants 20 years earlier and where soybeans were not grown for the next 17 years. The strains remained viable and symbiotically competent, and molecular and immunochemical methods showed that the strains were undistinguishable from the original inoculum strains USDA 110 and USDA 123. We also show that the strains had balanced numbers and their mobility in soil was low. To our knowledge, this is the first report showing the extra-long-term persistence of soybean-nodulating strains introduced into Polish soils and the first analyzing the long-term competitive relations of USDA 110 and USDA 123 after the two strains, neither of which was native, were introduced into the environment almost 2 decades ago.

  3. Chadong Block

    Institute of Scientific and Technical Information of China (English)

    1996-01-01

    @@ The Chadong Block, located in the east of Qaidam Basin, Qinghai Province, covers an area of 12 452 km2. It is bounded by Kunlum Mountains in the south and the northwest is closely adjacent to Aimunike Mountain.Rivers are widely distributed, which always run in NWSE direction, including the Sulunguole, Qaidam and Haluwusu Rivers. The traffic condition is good, the Qinghai-Tibet highway stretching through the whole area and the Lan-Qing railway, 20-50 km away from the block, passing from north to west. A lot of Mongolia minority people have settled there, of which herdsmen always live nearby the Qaidam River drainage area.

  4. Serodiagnosis of Schistosoma mansoni infections in an endemic area of Burkina Faso: performance of several immunological tests with different parasite antigens.

    Science.gov (United States)

    Sorgho, Hermann; Bahgat, Mahmoud; Poda, Jean-Noel; Song, Wenjian; Kirsten, Christa; Doenhoff, Michael J; Zongo, Issaka; Ouédraogo, Jean-Bosco; Ruppel, Andreas

    2005-02-01

    The performance of indirect haemagglutination assays (IHA), enzyme-linked immunosorbent assays (ELISA) and indirect immunofluorescent antibody tests (IFAT) were compared with 450 sera from a Schistosoma mansoni-endemic area in Burkina Faso. All participants in this survey provided at least one sample each of stool, urine and serum. From those with an egg-negative Kato-Katz thick smear, a second stool sample was examined. IHA was based on either extracts of adult S. mansoni worms (SmIHA) or S. japonicum egg antigen (SjIHA). For ELISA, three antigen preparations were used, namely: (i) soluble S. mansoni adult worm antigens (SWAP); (ii) soluble S. mansoni egg antigens (SEA); and (iii) a cationic exchange fraction of S. mansoni eggs (CEF6). IFAT was performed with S. mansoni male worm sections. Among the egg-excretors, the sensitivity of ELISA was high and egg antigens performed slightly better (SEA, 96%; CEF6, 97%) than worm antigen (94%). Sensitivity of IHA was satisfactory with homologous (Sm, >85%), but not heterologous (Sj, 56%) parasite antigen. In IFAT, the parenchyma-associated fluorescence showed high sensitivity (95%), but gut-associated fluorescence, which is known to be a sensitive diagnostic marker for schistosome-infected European travelers, was observed only in 76% of a sub-sample of 100 of the endemic sera. Among sera from egg-negative individuals, many gave positive reactions in several or all of the tests employed. These reactions (formally "false positive") are considered to represent true infections, since chemotherapy had not yet been delivered to this population. For the purpose of further surveys in Burkina Faso or other resource-poor settings, we suggest IHA as an accurate diagnostic test and propose to further improve its performance by including egg rather than worm antigens.

  5. Ultrastructural and biochemical detection of biotin and biotinylated polypeptides in Schistosoma mansoni

    Directory of Open Access Journals (Sweden)

    Santos P.R.P.

    1997-01-01

    Full Text Available Biotinylation is proposed for the identification of surface proteins in Schistosoma mansoni using the streptavidin-HRP conjugate for the detection of labeled polypeptides. However, control samples also showed several endogenous biotinylated polypeptides. In an attempt to determine the possibility of nonspecific binding between the streptavidin-HRP conjugate and polypeptides from S. mansoni, the conjugate was blocked with biotinamidecaproate-N-hydroxysuccinimide ester (BcapNHS before biotin-streptavidin blotting. No bands were detected on the nitrocellulose sheet, demonstrating the specific recognition of biotin by the streptavidin present in the conjugate. Whole cercariae and cercarial bodies and tails showed several endogenous biotinylated polypeptides. The biotin concentration was 13 µg/190,000 cercariae. Adult worms presented less endogenous biotinylated polypeptides than cercariae. These results may be due to changes in the environment from aerobic to anaerobic conditions when cercarial bodies (schistosomula are transformed into adult worms and a decrease in CO2 production may occur. Cercariae, cercarial bodies and adult male worms were examined by transmission electron microscopy employing an avidin-colloidal gold conjugate for the detection of endogenous biotin. Gold particles were distributed mainly on the muscle fibers, but dispersed granules were observed in the tegument, mitochondria and cytosol. The discovery of endogenous biotin in S. mansoni should be investigated in order to clarify the function of this vitamin in the parasite

  6. Detection of Schistosoma mansoni and Schistosoma haematobium by Real-Time PCR with High Resolution Melting Analysis.

    Science.gov (United States)

    Sady, Hany; Al-Mekhlafi, Hesham M; Ngui, Romano; Atroosh, Wahib M; Al-Delaimy, Ahmed K; Nasr, Nabil A; Dawaki, Salwa; Abdulsalam, Awatif M; Ithoi, Init; Lim, Yvonne A L; Chua, Kek Heng; Surin, Johari

    2015-07-16

    The present study describes a real-time PCR approach with high resolution melting-curve (HRM) assay developed for the detection and differentiation of Schistosoma mansoni and S. haematobium in fecal and urine samples collected from rural Yemen. The samples were screened by microscopy and PCR for the Schistosoma species infection. A pair of degenerate primers were designed targeting partial regions in the cytochrome oxidase subunit I (cox1) gene of S. mansoni and S. haematobium using real-time PCR-HRM assay. The overall prevalence of schistosomiasis was 31.8%; 23.8% of the participants were infected with S. haematobium and 9.3% were infected with S. mansoni. With regards to the intensity of infections, 22.1% and 77.9% of S. haematobium infections were of heavy and light intensities, respectively. Likewise, 8.1%, 40.5% and 51.4% of S. mansoni infections were of heavy, moderate and light intensities, respectively. The melting points were distinctive for S. mansoni and S. haematobium, categorized by peaks of 76.49 ± 0.25 °C and 75.43 ± 0.26 °C, respectively. HRM analysis showed high detection capability through the amplification of Schistosoma DNA with as low as 0.0001 ng/µL. Significant negative correlations were reported between the real-time PCR-HRM cycle threshold (Ct) values and microscopic egg counts for both S. mansoni in stool and S. haematobium in urine (p < 0.01). In conclusion, this closed-tube HRM protocol provides a potentially powerful screening molecular tool for the detection of S. mansoni and S. haematobium. It is a simple, rapid, accurate, and cost-effective method. Hence, this method is a good alternative approach to probe-based PCR assays.

  7. Detection of Schistosoma mansoni and Schistosoma haematobium by Real-Time PCR with High Resolution Melting Analysis

    Directory of Open Access Journals (Sweden)

    Hany Sady

    2015-07-01

    Full Text Available The present study describes a real-time PCR approach with high resolution melting-curve (HRM assay developed for the detection and differentiation of Schistosoma mansoni and S. haematobium in fecal and urine samples collected from rural Yemen. The samples were screened by microscopy and PCR for the Schistosoma species infection. A pair of degenerate primers were designed targeting partial regions in the cytochrome oxidase subunit I (cox1 gene of S. mansoni and S. haematobium using real-time PCR-HRM assay. The overall prevalence of schistosomiasis was 31.8%; 23.8% of the participants were infected with S. haematobium and 9.3% were infected with S. mansoni. With regards to the intensity of infections, 22.1% and 77.9% of S. haematobium infections were of heavy and light intensities, respectively. Likewise, 8.1%, 40.5% and 51.4% of S. mansoni infections were of heavy, moderate and light intensities, respectively. The melting points were distinctive for S. mansoni and S. haematobium, categorized by peaks of 76.49 ± 0.25 °C and 75.43 ± 0.26 °C, respectively. HRM analysis showed high detection capability through the amplification of Schistosoma DNA with as low as 0.0001 ng/µL. Significant negative correlations were reported between the real-time PCR-HRM cycle threshold (Ct values and microscopic egg counts for both S. mansoni in stool and S. haematobium in urine (p < 0.01. In conclusion, this closed-tube HRM protocol provides a potentially powerful screening molecular tool for the detection of S. mansoni and S. haematobium. It is a simple, rapid, accurate, and cost-effective method. Hence, this method is a good alternative approach to probe-based PCR assays.

  8. An iron uptake operon required for proper nodule development in the Bradyrhizobium japonicum-soybean symbiosis.

    Science.gov (United States)

    Benson, Heather P; Boncompagni, Eric; Guerinot, Mary Lou

    2005-09-01

    Rhizobia live in the soil or enter into a nitrogen-fixing symbiosis with a suitable host plant. Each environment presents different challenges with respect to iron acquisition. The soybean symbiont Bradyrhizobium japonicum 61A152 can utilize a variety of siderophores (Fe[III]-specific ligands). Purification of iron-regulated outer membrane proteins had previously allowed the cloning of a gene, fegA, from B. japonicum 61A152, whose predicted protein shares significant amino acid similarity with known TonB-dependent siderophore receptors. Here, we show that fegA is in an operon with a gene, fegB, that is predicted to encode an inner membrane protein. Characterization of fegAB and fegB mutants shows that bothfegA and fegB are required for utilization of the siderophore ferrichrome. Whereas thefegB mutant forms a normal symbiosis, the fegAB mutant has a dramatic phenotype in planta. Six weeks after inoculation with a fegAB strain, soybean nodules do not contain leghemoglobin and do not fix nitrogen. Infected cells contain few symbiosomes and are filled with vesicles. As ferrichrome is a fungal siderophore not likely to be available in nodules, the symbiotic defect suggests that the fegAB operon is serving a different function in planta, possibly one involved in signaling between the two partners.

  9. Differentiation of Cirsium japonicum and C. setosum by TLC and HPLC-MS.

    Science.gov (United States)

    Ganzera, Markus; Pöcher, Astrid; Stuppner, Hermann

    2005-01-01

    The Chinese Pharmacopoeia indicates the use of field thistle (Cirsium setosum) and Japanese field thistle (C. japonicum) in the treatment of bleeding and inflammation. In the absence of an analytical method for the differentiation and analysis of these two species, TLC and HPLC-MS methods have been developed for this purpose. Both species could be readily distinguished by their flavonoid pattern as revealed by TLC on silica gel layers eluted with ethyl acetate:formic acid:acetic acid:water. The quantitative determination of four flavonoids, namely hispidulin-7-neohesperidoside, linarin, pectolinarin and luteolin, was possible using HPLC. Their optimum separation was achieved on a C12 column eluted with water and 0.025% trifluoroacetic acid in acetonitrile. HPLC-MS experiments were performed to confirm peak identity. In samples of C. japonicum, pectolinarin was the major flavonoid (0.32-2.00%), followed by linarin, hispidulin-7-neohesperidoside and luteolin; the total flavonoid content varied from 0.81 to 3.67%. In C. setosum only one flavonoid (linarin; 1.36-2.83%) was assignable. The HPLC method was validated for linearity, limit of detection (< or = 1.7 ng on-column), peak purity, repeatability (< or = 2.3%) and accuracy (recovery rates of spiked samples were between 99.2 and 101.6%).

  10. (Iron regulation of gene expression in the Bradyrhizobium japonicum/soybean symbiosis)

    Energy Technology Data Exchange (ETDEWEB)

    Guerinot, M.L.

    1992-01-01

    We wish to address the question of whether iron plays a regulatory role in the Bradyrhizobium japonicum/soybeam symbiosis. Iron may be an important regulatory signal in planta as the bacteria must acquire iron from their plant hosts and iron-containing proteins figure prominently in all nitrogen-fixing symbioses. For example, the bacterial partner is believed to synthesize the heme moiety of leghemoglobin, which may represent as much as 25--30% of the total soluble protein in an infected plant cell. For this reason, we have focused our attention on the regulation by iron of the first step in the bacterial heme biosynthetic pathway. The enzyme which catalyzes this step, 5-aminolevulinic acid synthase, is encoded by the hemA gene which we had previously cloned and sequenced. Specific objectives include: to define the cis-acting sequences which confer iron regulation on the B. japonicum hemA gene; to identify trans-acting factors which regulate the expression of hemA by iron; to identify new loci which are transcriptionally responsive to changes in iron availability; and to examine the effects of mutations in various known regulatory genes for their effect on the expression of hemA.

  11. [Iron regulation of gene expression in the Bradyrhizobium japonicum/soybean symbiosis]. Progress report

    Energy Technology Data Exchange (ETDEWEB)

    Guerinot, M.L.

    1992-06-01

    We wish to address the question of whether iron plays a regulatory role in the Bradyrhizobium japonicum/soybeam symbiosis. Iron may be an important regulatory signal in planta as the bacteria must acquire iron from their plant hosts and iron-containing proteins figure prominently in all nitrogen-fixing symbioses. For example, the bacterial partner is believed to synthesize the heme moiety of leghemoglobin, which may represent as much as 25--30% of the total soluble protein in an infected plant cell. For this reason, we have focused our attention on the regulation by iron of the first step in the bacterial heme biosynthetic pathway. The enzyme which catalyzes this step, 5-aminolevulinic acid synthase, is encoded by the hemA gene which we had previously cloned and sequenced. Specific objectives include: to define the cis-acting sequences which confer iron regulation on the B. japonicum hemA gene; to identify trans-acting factors which regulate the expression of hemA by iron; to identify new loci which are transcriptionally responsive to changes in iron availability; and to examine the effects of mutations in various known regulatory genes for their effect on the expression of hemA.

  12. Mitigation of nitrous oxide emissions from soils by Bradyrhizobium japonicum inoculation

    Science.gov (United States)

    Itakura, Manabu; Uchida, Yoshitaka; Akiyama, Hiroko; Hoshino, Yuko Takada; Shimomura, Yumi; Morimoto, Sho; Tago, Kanako; Wang, Yong; Hayakawa, Chihiro; Uetake, Yusuke; Sánchez, Cristina; Eda, Shima; Hayatsu, Masahito; Minamisawa, Kiwamu

    2013-03-01

    Nitrous oxide (N2O) is a greenhouse gas that is also capable of destroying the ozone layer. Agricultural soil is the largest source of N2O (ref. ). Soybean is a globally important leguminous crop, and hosts symbiotic nitrogen-fixing soil bacteria (rhizobia) that can also produce N2O (ref. ). In agricultural soil, N2O is emitted from fertilizer and soil nitrogen. In soybean ecosystems, N2O is also emitted from the degradation of the root nodules. Organic nitrogen inside the nodules is mineralized to NH4+, followed by nitrification and denitrification that produce N2O. N2O is then emitted into the atmosphere or is further reduced to N2 by N2O reductase (N2OR), which is encoded by the nosZ gene. Pure culture and vermiculite pot experiments showed lower N2O emission by nosZ+ strains and nosZ++ strains (mutants with increased N2OR activity) of Bradyrhizobium japonicum than by nosZ- strains. A pot experiment using soil confirmed these results. Although enhancing N2OR activity has been suggested as a N2O mitigation option, this has never been tested in the field. Here, we show that post-harvest N2O emission from soybean ecosystems due to degradation of nodules can be mitigated by inoculation of nosZ+ and non-genetically modified organism nosZ++ strains of B. japonicum at a field scale.

  13. Semen quality in Schistosoma haematobium infected men in Madagascar.

    Science.gov (United States)

    Leutscher, Peter D C; Høst, Erik; Reimert, Claus M

    2009-01-01

    The seminal vesicles and the prostate are frequently affected by egg-induced inflammation in Schistosoma haematobium infected men. The objective of this study was to assess the semen quality in men with male genital schistosomiasis (MGS). The examination of the semen samples was performed in men aged 15 to 49 years living an S. haematobium endemic area in Madagascar prior to anti-schistosoma treatment with praziquantel and five months later. Men from the high endemic Sirama sugarcane plantation with positive egg excretion in the urine and circulating anodic antigen (CAA) present in serum (n=85) were compared to men (without egg excretion and no CAA) from the neighbouring low-endemic Mataipako village (n=57). The proportion of men with egg excretion in semen was significantly higher in Sirama than in Mataipako (53% versus 4%), whereas the median ejaculate volume was significantly lower in Sirama (1.8 ml versus 2.4 ml). There was no statistical difference in median spermatocyte counts and in the proportions of men detected with azoospermia. The mean apoptotic rate was 7.8% in a subgroup of 30 men from Sirama. A positive correlation was found between apoptotic rate and seminal eosinophil cationic protein (ECP) level (rho=0.560; Pinfection is associated with sperm apoptosis and a reduced production of seminal fluid. Egg induced inflammation in the seminal vesicles and the prostate could be underlying mechanism for both observations.

  14. THE SUSCEPTIBILITY OF RECENT ISOLATES OF Schistosoma mansoni TO PRAZIQUANTEL

    Directory of Open Access Journals (Sweden)

    Adriana Maria B. MENDONÇA

    2016-01-01

    Full Text Available Introduction: Schistosomiasis is a chronic disease caused by trematode flatworms of the genus Schistosoma and its control is dependent on a single drug, praziquantel (PZQ, but concerns over PZQ resistance have renewed interest in evaluating the in vitro susceptibility of recent isolates of Schistosoma mansoni to PZQ in comparison with well-established strains in the laboratory. Material and methods: The in vitro activity of PZQ (6.5-0.003 µg/mL was evaluated in terms of mortality, reduced motor activity and ultrastructural alterations against S. mansoni. Results: After 3 h of incubation, PZQ, at 6.5 µg/mL, caused 100% mortality of all adult worms in the three types of recent isolates, while PZQ was inactive at concentrations of 0.08-0.003 µg/mL after 3 h of incubation. The results show that the SLM and Sotave isolates basically presented the same pattern of susceptibility, differing only in the concentration of 6.5 µg/mL, where deaths occurred from the range of 1.5 h in Sotave and just in the 3 h range of SLM. Additionally, this article presents ultrastructural evidence of rapid severe PZQ-induced surface membrane damage in S. mansoni after treatment with the drug, such as disintegration, sloughing, and erosion of the surface. Conclusion: According to these results, PZQ is very effective to induce tegument destruction of recent isolates of S. mansoni.

  15. Effects of Cu on the content of chlorophylls and secondary metabolites in the Cu-hyperaccumulator lichen Stereocaulon japonicum.

    Science.gov (United States)

    Nakajima, Hiromitsu; Hara, Kojiro; Yamamoto, Yoshikazu; Itoh, Kiminori

    2015-03-01

    Understanding the relationship between Cu and Cu-hyperaccumulator lichens is important for their application in monitoring and assessing heavy metal pollution. We investigated the Cu-hyperaccumulator lichen Stereocaulon japonicum at several Cu-polluted and control sites in Japan, and found the lichen to be widely distributed. Its concentrations of Cu, chlorophylls, and secondary metabolites, chlorophyll-related indices, and absorption spectra were measured, and we observed negative effects of Cu on these concentrations and indices. For highly Cu-polluted samples (>100ppm dry weight), however, we found significant linear correlations between Cu and chlorophyll concentrations. This can be considered as the response of the photobiont in S. japonicum to Cu stress. In highly Cu-polluted samples the chlorophyll-related indices and concentration of total secondary metabolites were almost constant regardless of Cu concentration. This suggests that the increase in chlorophyll concentration with the increase in Cu concentration enhances photosynthetic productivity per unit biomass, which will allow the production of extra structure and energy for maintaining the chlorophyll-related indices under Cu stress. The relationship between the increase in chlorophyll concentration of S. japonicum and the decrease in secondary metabolite concentration of the lichen can be explained by considering the balance of carbohydrates in the lichen. We found that a spectral index A372-A394 can be a useful index of the concentrations of Cu and total secondary metabolites in S. japonicum. These findings show the adjustment of the content of chlorophylls and secondary metabolites in S. japonicum to Cu stress, and provide a better understanding of the relationship between Cu and the Cu-hyperaccumulator lichen.

  16. Identification of the Schistosoma mansoni Molecular Target for the Antimalarial Drug Artemether

    KAUST Repository

    Lepore, Rosalba

    2011-11-28

    Plasmodium falciparum and Schistosoma mansonii are the parasites responsible for most of the malaria and schistosomiasis cases in the world. Notwithstanding their many differences, the two agents have striking similarities in that they both are blood feeders and are targets of an overlapping set of drugs, including the well-known artemether molecule. Here we explore the possibility of using the known information about the mode of action of artemether in Plasmodium to identify the molecular target of the drug in Schistosoma and provide evidence that artemether binds to SmSERCA, a putative Ca2+-ATPase of Schistosoma. We also predict the putative binding mode of the molecule for both its Plasmodium and Schistosoma targets. Our analysis of the mode of binding of artemether to Ca2+-ATPases also provides an explanation for the apparent paradox that, although the molecule has no side effect in humans, it has been shown to possess antitumoral activity. © 2011 American Chemical Society.

  17. The bladder carcinoma secondary to schistosoma mansoni infection: A case report with review of the literature

    Directory of Open Access Journals (Sweden)

    Murat Can Kiremit

    2015-01-01

    Conclusion: Regarding the increasing travels all around the world, clinicians should remember that Schistosoma infection is certainly a part of the differential diagnosis of bladder carcinoma, even if the patients are not from endemic regions.

  18. Schistosome syntenin partially protects vaccinated mice against Schistosoma mansoni infection.

    Directory of Open Access Journals (Sweden)

    Barbara C Figueiredo

    2014-08-01

    Full Text Available Schistosomiasis is a neglected tropical disease caused by several species of trematode of the genus Schistosoma. The disease affects more than 200 million people in the world and causes up to 280,000 deaths per year, besides having high morbidity due to chronic illness that damages internal organs. Current schistosomiasis control strategies are mainly based on chemotherapy, but many researchers believe that the best long-term strategy to control disease is a combination of drug treatment and immunization with an anti-schistosome vaccine. Among the most promising molecules as vaccine candidates are the proteins present in the tegument and digestive tract of the parasite.In this study, we describe for the first time Schistosoma mansoni syntenin (SmSynt and we evaluate its potential as a recombinant vaccine. We demonstrate by real-time PCR that syntenin is mainly expressed in intravascular life stages (schistosomula and adult worms of the parasite life cycle and, by confocal microscopy, we localize it in digestive epithelia in adult worms and schistosomula. Administration of siRNAs targeting SmSynt leads to the knock-down of syntenin gene and protein levels, but this has no demonstrable impact on parasite morphology or viability, suggesting that high SmSynt gene expression is not essential for the parasites in vitro. Mice immunization with rSmSynt, formulated with Freund's adjuvant, induces a Th1-type response, as suggested by the production of IFN-γ and TNF-α by rSmSynt-stimulated cultured splenocytes. The protective effect conferred by vaccination with rSmSynt was demonstrated by 30-37% reduction of worm burden, 38-43% reduction in the number, and 35-37% reduction in the area, of liver granulomas.Our report is the first characterization of syntenin in Schistosoma mansoni and our data suggest that this protein is a potential candidate for the development of a multi-antigen vaccine to control schistosomiasis.

  19. Condiloma acuminado anal com ovos de Schistosoma mansoni em paciente HIV-positivo Condilomata acuminata with Schistosoma eggs in HIV-positive patient

    Directory of Open Access Journals (Sweden)

    Fabiana Pirani Carneiro

    2007-06-01

    Full Text Available Condiloma acuminado e ovos de Schistosoma são freqüentemente encontrados na região anal, mas não há nenhum caso descrito de associação dessas doenças nessa região. No colo uterino a associação de infecção por HPV (vírus do papiloma humano e ovos de Schistosoma em paciente HIV (vírus da imunodeficiência humana-positivo já foi relatada e há evidências de que essa associação possa alterar a história natural dessas doenças. Assim como no colo uterino, é possível que essa interação também ocorra na região anal. Nosso objetivo, portanto, é relatar um caso de condiloma anal em paciente HIV-positivo, que foi submetido a ressecção cirúrgica e que apresentou no exame histopatológico numerosos ovos de Schistosoma mansoni.Condilomata acuminata and Schistosoma eggs are frequently found in the anal region, but there is no report about the association of these diseases in this region. The association of HPV infection and Schistosoma eggs in an HIV-positive patient was found in uterine cervix and there is evidence suggesting that this association can alters the natural history of these diseases. Like in the cervix, it is possible that this interaction also occurs in the anal region. So, we report a case of anal Condilomata acuminata, in an HIV-positive patient, that was ressected and contained on histopathologic examination, multiple Schistosoma eggs.

  20. The effect of Zymomonas mobilis culture on experimental Schistosoma mansoni infection O efeito da cultura de Zymomonas mobilis na infecção experimental por Schistosoma mansoni

    Directory of Open Access Journals (Sweden)

    Juliana de Fátima Macedo Santos

    2004-12-01

    Full Text Available C57Bl/10 male mice infected with Schistosoma mansoni were distributed into mixed, prophylactic and curative groups. A culture of Zymomonas mobilis was orally administered to mice. A 61% protection from the infection was observed in the curative group (p Camundongos C57Bl/10 do sexo masculino, infectados com Schistosoma mansoni foram distribuídos nos grupos misto, profilático e curativo. Cultura de Zymomonas mobilis foi administrada oralmente aos camundongos. Uma proteção de 61% foi observada no grupo curativo (p<0,05. Os estudos histopatológicos dos fígados e intestinos mostraram resultados similares.

  1. [Effect of plant growth regulators on physiological activity of Bradyrhizobium japonicum ].

    Science.gov (United States)

    Leonova, N O; Tytova, L V; Tantsiurenko, O V; Antypchuk, A F

    2005-01-01

    Influence of plant growth regulators Ivin, Emistim C, Eney and Agrostimulin on the biomass production and exopolymers synthesis of soybean nodule bacteria, which have contrasting symbiotic properties, and glutamine synthetase activity of their cell-free extracts were studied. It was shown that the processes of the biomass and exopolymers accumulation had an opposite direction. Of all preparations only Ivin and Agrostimulin intensificol growth activity of the microorganisms under study. The level of glutamine synthetase activity and this enzymatic reaction specificity to the bivalent metal ions were determined by the special features of Bradyrhizobium strains and nature of the plant growth regulators. Only in the presence of Eney the increase of glutamine synthetase activity of both cultures of Bradyrhizobium japonicum was established.

  2. Cercarial emergence pattern of Schistosoma haematobium from Libreville, Gabon

    Directory of Open Access Journals (Sweden)

    Mintsa-Nguéma Rodrigue

    2014-01-01

    Full Text Available Although schistosomiasis has been a public health issue in Gabon for nearly a century, little is known about its current transmission dynamics. We analyzed the chronobiology of 137 cercarial emission profiles of Schistosoma haematobium from Libreville, the capital of Gabon, located in an open area for schistosomiasis. We found that 88% of the cercariae were shed between 11 a.m. and 3 p.m. and that the average pattern was of circadian type, with the average peak at 1 p.m., and representing 27% of the total number of cercariae of the day. The rhythms of emergence may be associated with environmental pressures on the parasite, especially those related to their definitive hosts.

  3. Schistosoma mansoni cercariae exploit an elastohydrodynamic coupling to swim efficiently

    CERN Document Server

    Krishnamurthy, Deepak; Bhargava, Arjun; Prakash, Manu

    2016-01-01

    The motility of many parasites is critical for the infection process of their host, as exemplified by the transmission cycle of the blood fluke Schistosoma mansoni. In their human infectious stage, immature, submillimetre-scale forms of the parasite known as cercariae swim in freshwater and infect humans by penetrating through the skin. This infection causes Schistosomiasis, a parasitic disease that is comparable to malaria in its global socio-economic impact. Given that cercariae do not feed and hence have a finite lifetime of around 12 hours, efficient motility is crucial for the parasite's survival and transmission of Schistosomiasis. However, a first-principles understanding of how cercariae swim is lacking. Via a combined experimental, theoretical and robotics based approach, we demonstrate that cercariae propel themselves against gravity by exploiting a unique elastohydrodynamic coupling. We show that cercariae beat their tail in a periodic fashion while maintaining a fixed flexibility near their poster...

  4. Cercarial emergence pattern of Schistosoma haematobium from Libreville, Gabon.

    Science.gov (United States)

    Mintsa-Nguéma, Rodrigue; Moné, Hélène; Ibikounlé, Moudachirou; Mengué-Ngou-Milama, Krystina; Kombila, Maryvonne; Mouahid, Gabriel

    2014-01-01

    Although schistosomiasis has been a public health issue in Gabon for nearly a century, little is known about its current transmission dynamics. We analyzed the chronobiology of 137 cercarial emission profiles of Schistosoma haematobium from Libreville, the capital of Gabon, located in an open area for schistosomiasis. We found that 88% of the cercariae were shed between 11 a.m. and 3 p.m. and that the average pattern was of circadian type, with the average peak at 1 p.m., and representing 27% of the total number of cercariae of the day. The rhythms of emergence may be associated with environmental pressures on the parasite, especially those related to their definitive hosts.

  5. Estudios inmunologicos en hamsters (Cricetus auratus infectados con Schistosoma mansoni

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    Eduardo Monge

    1986-08-01

    Full Text Available Los resultados de este trabajo muestran que el hamster (Cricetus auratus puede ser utilizado como un modelo experimental para estudios inmunológicos en la infección por Schistosoma mansoni. Los datos obtenidos, relativos a inmunidad concomitante, producción de anticuerpo letal e inmunosupresión se asemejan a los conseguidos en otros modelos experimentales ya establecidos. Estas observaciones indican que el hámster, además de ser un hospedero satisfactorio para el mantenimiento del parásito en el laboratorio, puede ser considerado como un modelo experimental alterno cuyo crecimiento y mantenimiento son relativamente simples y además es un animal de fácil manejo.

  6. A potential vector of Schistosoma mansoni in Uruguay Um vetor potencial do Schistosoma mansoni no Uruguai

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    W. Lobato Paraense

    1989-09-01

    Full Text Available Susceptibily experiments were carried out with a Biomphalaria straminea-like planorbid snail (Biomphalaria aff. straminea, species inquirenda from Espinillar, near Salto (Uruguay, in the area of the Salto Grande reservoir, exposed individually to 5 miracidia of Schistosoma mansoni (SJ2 and BH2 strains. Of 130 snails exposed to the SJ2 strain, originally infective to Biomphalaria tenagophila, 30 became infected (23%. The prepatent (precercaria period ranged from 35 to 65 days. The cercarial output was irregular, following no definite pattern, varying from 138 to 76,075 per snail (daily average 4.3 to 447.5 and ending up with death. Three specimens that died, without having shed cercarie, on days 69 (2 and 80 after exposure to miracidia, had developing secondary sporocysts in their tissues, justifying the prospect of a longer precercarial period in these cases. In a control group of 120 B. teangophila, exposed to the SJ2 strain, 40 became infected, showing an infection rate (33.3% not significantly different from that of the Espinillar snail (X [raised to the power of] 2 = 3.26. No cercarie were produced by any of the Espinilar snails exposed to miracidia of the BH2 strain, originally infective to Biomphalaria glabrata. Four specimens showed each a primary sporocyst in one tentacle, which disappeared between 15 and 25 days post-exposure, and two others died with immature, very slender sporocysts in their tissues on days 36 and 54. In a control group of 100 B. glabrata exposed to BH2 miracidia, 94 shed cercariae (94% and 6 remained negative. Calculation of Frandsen's (1979a, b TCP/100 index shows that "Espinillar Biomphalaria-SJ2 S. mansoni" is a vector-parasite "compatible" combination. Seeing that tenagophila-borne schistosomiasis is prevalent in Rio de Janeiro and São Paulo states and has recently spread sothwards to Santa Catarina state, and the range of B. tenagophila overlaps taht of the Espinillar Biomphalaria, the possibility of

  7. Characterization of human heterophil hemolysins induced by Schistosoma mansoni infection

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    Munir Chamone

    1983-09-01

    Full Text Available Heterophil antibodies could be detected in sera from normal or from patient with chronic schistosomiasis. Their hemolytic activities depend on the integrity of the complement classic pathway. The heterophil antibodies from patient sera presented a higher specificity for Schistosoma mansoni antigen preparations than those detected in normal sera. Most of the hemolytic activity observed in normal sera can be destroyed at 56ºC for 4 min. On the other hand, about 80% of the sera from infected patients are partially or totally resistant to this heat-treatment. The hemolytic activities of sera were eluted from a gel filtration column in different fractions of the first peak.Anticorpos heterófilos foram detectados nos soros de pacientes normais ou com esquistossomose mansoni crônica. Suas atividades hemolíticas dependem da integridade da via clássica do sistema do complemento. Os anticorpos heterófilos dos pacientes esquistossomóticos apresentaram maior especificidade para antígenos de Schistosoma mansoni do que aqueles anticorpos detectados nos soros de pacientes normais. A atividade hemolítica do anticorpo nos soros normais podia ser destruída pelo aquecimento destes soros a 56ºC durante 4 minutos. Por outro lado, cerca de 80% dos soros de pacientes esquistossomóticos eram parcial ou totalmente resistentes ao mesmo tratamento. As atividades dos anticorpos heterófilos foram eluídas através da filtração em gel, em diferentes frações no primeiro pico.

  8. Quantitative Phosphoproteomic Analysis of Soybean Root Hairs Inoculated with Bradyrhizobium japonicum

    Energy Technology Data Exchange (ETDEWEB)

    Nguyen, Tran H.; Brechenmacher, Laurent; Aldrich, Joshua T.; Clauss, Therese RW; Gritsenko, Marina A.; Hixson, Kim K.; Libault, Marc; Tanaka, Kiwamu; Yang, Feng; Yao, Qiuming; Pasa-Tolic, Ljiljana; Xu, Dong; Nguyen, Henry T.; Stacey, Gary

    2012-11-11

    Root hairs are single hair-forming cells on roots that function to increase root surface area, enhancing water and nutrient uptake. In leguminous plants, root hairs also play a critical role as the site of infection by symbiotic nitrogen fixing rhizobia, leading to the formation of a novel organ, the nodule. The initial steps in the rhizobia-root hair infection process are known to involve specific receptor kinases and subsequent kinase cascades. Here, we characterize the phosphoproteome of the root hairs and the corresponding stripped roots (i.e., roots from which root hairs were removed) during rhizobial colonization and infection to gain insight into the molecular mechanism of root hair cell biology. We chose soybean (Glycine max L.), one of the most important crop plants in the legume family, for this study because of its larger root size, which permits isolation of sufficient root hair material for phosphoproteomic analysis. Phosphopeptides derived from root hairs and stripped roots, mock inoculated or inoculated with the soybean-specific rhizobium Bradyrhizobium japonicum, were labeled with the isobaric tag 8-plex ITRAQ, enriched using Ni-NTA magnetic beads and subjected to nRPLC-MS/MS analysis using HCD and decision tree guided CID/ETD strategy. A total of 1,625 unique phosphopeptides, spanning 1,659 non-redundant phosphorylation sites, were detected from 1,126 soybean phosphoproteins. Among them, 273 phosphopeptides corresponding to 240 phosphoproteins were found to be significantly regulated (>1.5 fold abundance change) in response to inoculation with B. japonicum. The data reveal unique features of the soybean root hair phosphoproteome, including root hair and stripped root-specific phosphorylation suggesting a complex network of kinase-substrate and phosphatase-substrate interactions in response to rhizobial inoculation.

  9. Quantitative Phosphoproteomic Analysis of Soybean Root Hairs Inoculated with Bradyrhizobium japonicum*

    Science.gov (United States)

    Nguyen, Tran Hong Nha; Brechenmacher, Laurent; Aldrich, Joshua T.; Clauss, Therese R.; Gritsenko, Marina A.; Hixson, Kim K.; Libault, Marc; Tanaka, Kiwamu; Yang, Feng; Yao, Qiuming; Paša-Tolić, Ljiljana; Xu, Dong; Nguyen, Henry T.; Stacey, Gary

    2012-01-01

    Root hairs are single hair-forming cells on roots that function to increase root surface area, enhancing water and nutrient uptake. In leguminous plants, root hairs also play a critical role as the site of infection by symbiotic nitrogen fixing rhizobia, leading to the formation of a novel organ, the nodule. The initial steps in the rhizobia-root hair infection process are known to involve specific receptor kinases and subsequent kinase cascades. Here, we characterize the phosphoproteome of the root hairs and the corresponding stripped roots (i.e. roots from which root hairs were removed) during rhizobial colonization and infection to gain insight into the molecular mechanism of root hair cell biology. We chose soybean (Glycine max L.), one of the most important crop plants in the legume family, for this study because of its larger root size, which permits isolation of sufficient root hair material for phosphoproteomic analysis. Phosphopeptides derived from root hairs and stripped roots, mock inoculated or inoculated with the soybean-specific rhizobium Bradyrhizobium japonicum, were labeled with the isobaric tag eight-plex iTRAQ, enriched using Ni-NTA magnetic beads and subjected to nanoRPLC-MS/MS1 analysis using HCD and decision tree guided CID/ETD strategy. A total of 1625 unique phosphopeptides, spanning 1659 nonredundant phosphorylation sites, were detected from 1126 soybean phosphoproteins. Among them, 273 phosphopeptides corresponding to 240 phosphoproteins were found to be significantly regulated (>1.5-fold abundance change) in response to inoculation with B. japonicum. The data reveal unique features of the soybean root hair phosphoproteome, including root hair and stripped root-specific phosphorylation suggesting a complex network of kinase-substrate and phosphatase-substrate interactions in response to rhizobial inoculation. PMID:22843990

  10. Quantitative phosphoproteomic analysis of soybean root hairs inoculated with Bradyrhizobium japonicum.

    Science.gov (United States)

    Nguyen, Tran Hong Nha; Brechenmacher, Laurent; Aldrich, Joshua T; Clauss, Therese R; Gritsenko, Marina A; Hixson, Kim K; Libault, Marc; Tanaka, Kiwamu; Yang, Feng; Yao, Qiuming; Pasa-Tolić, Ljiljana; Xu, Dong; Nguyen, Henry T; Stacey, Gary

    2012-11-01

    Root hairs are single hair-forming cells on roots that function to increase root surface area, enhancing water and nutrient uptake. In leguminous plants, root hairs also play a critical role as the site of infection by symbiotic nitrogen fixing rhizobia, leading to the formation of a novel organ, the nodule. The initial steps in the rhizobia-root hair infection process are known to involve specific receptor kinases and subsequent kinase cascades. Here, we characterize the phosphoproteome of the root hairs and the corresponding stripped roots (i.e. roots from which root hairs were removed) during rhizobial colonization and infection to gain insight into the molecular mechanism of root hair cell biology. We chose soybean (Glycine max L.), one of the most important crop plants in the legume family, for this study because of its larger root size, which permits isolation of sufficient root hair material for phosphoproteomic analysis. Phosphopeptides derived from root hairs and stripped roots, mock inoculated or inoculated with the soybean-specific rhizobium Bradyrhizobium japonicum, were labeled with the isobaric tag eight-plex iTRAQ, enriched using Ni-NTA magnetic beads and subjected to nanoRPLC-MS/MS1 analysis using HCD and decision tree guided CID/ETD strategy. A total of 1625 unique phosphopeptides, spanning 1659 nonredundant phosphorylation sites, were detected from 1126 soybean phosphoproteins. Among them, 273 phosphopeptides corresponding to 240 phosphoproteins were found to be significantly regulated (>1.5-fold abundance change) in response to inoculation with B. japonicum. The data reveal unique features of the soybean root hair phosphoproteome, including root hair and stripped root-specific phosphorylation suggesting a complex network of kinase-substrate and phosphatase-substrate interactions in response to rhizobial inoculation.

  11. A cyclohexanecarboxamide derivative with inhibitory effects on Schistosoma mansoni cercarial serine protease and penetration of mice skin by the parasite.

    Science.gov (United States)

    Bahgat, Mahmoud; Aboul-Enein, Mohamed N; El Azzouny, Aida A; Maghraby, Amany; Ruppel, Andreas; Soliman, Wael M

    2009-01-01

    A cyclohexanecarboxamide derivative, N-phenyl-N-[1-(piperidine-1-carbonyl)cyclohexyl] benzamide (MNRC-5), was evaluated for its inhibitory effects on Schistosoma mansoni cercarial serine protease activity and cercarial penetration. MNRC-5 exerted an inhibitory effect on S. mansoni cercarial serine protease at serial concentrations of the specific chromogenic substrate Boc-Val-Leu-Gly-Arg-PNA for such enzyme family and the inhibitory coefficient (Ki) value was deduced. Moreover, topical treatment of mice tails with the most potent inhibitory concentration of MNRC-5 formulated in jojoba oil successfully blocked cercarial penetration as demonstrated by a significant reduction (75%; p jojoba oil base containing no MNRC-5. In addition, the IgM and IgG reactivities to crude S. mansoni cercarial, worm and egg antigens were generally lower in sera from treated infected mice than untreated infected mice. In conclusion, we report on a new serine protease inhibitor capable for blocking penetration of host skin by S. mansoni cercariae as measured by lowering worm burden and decrease in the levels of both IgM and IgG towards different bilharzial antigens upon topical treatment.

  12. Evidence for Novel Pharmacological Sensitivities of Transient Receptor Potential (TRP Channels in Schistosoma mansoni.

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    Swarna Bais

    2015-12-01

    Full Text Available Schistosomiasis, caused by parasitic flatworms of the genus Schistosoma, is a neglected tropical disease affecting hundreds of millions globally. Praziquantel (PZQ, the only drug currently available for treatment and control, is largely ineffective against juvenile worms, and reports of PZQ resistance lend added urgency to the need for development of new therapeutics. Ion channels, which underlie electrical excitability in cells, are validated targets for many current anthelmintics. Transient receptor potential (TRP channels are a large family of non-selective cation channels. TRP channels play key roles in sensory transduction and other critical functions, yet the properties of these channels have remained essentially unexplored in parasitic helminths. TRP channels fall into several (7-8 subfamilies, including TRPA and TRPV. Though schistosomes contain genes predicted to encode representatives of most of the TRP channel subfamilies, they do not appear to have genes for any TRPV channels. Nonetheless, we find that the TRPV1-selective activators capsaicin and resiniferatoxin (RTX induce dramatic hyperactivity in adult worms; capsaicin also increases motility in schistosomula. SB 366719, a highly-selective TRPV1 antagonist, blocks the capsaicin-induced hyperactivity in adults. Mammalian TRPA1 is not activated by capsaicin, yet knockdown of the single predicted TRPA1-like gene (SmTRPA in S. mansoni effectively abolishes capsaicin-induced responses in adult worms, suggesting that SmTRPA is required for capsaicin sensitivity in these parasites. Based on these results, we hypothesize that some schistosome TRP channels have novel pharmacological sensitivities that can be targeted to disrupt normal parasite neuromuscular function. These results also have implications for understanding the phylogeny of metazoan TRP channels and may help identify novel targets for new or repurposed therapeutics.

  13. Comparación entre coinoculación con Bradyrhizobium japonicum y Azospirillum brasilense e inoculación simple con Bradyrhizobium japonicum en la nodulación, crecimiento y acumulación de N en el cultivo de soja Comparison of coinoculation with Bradyrhizobium japonicum and Azospirillum brasilense with inoculation with Bradyrhizobium japonicum on nodulation, growth, and N accumulation in the soybean crop

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    S. Benintende

    2010-12-01

    Full Text Available El objetivo fue comparar la coinoculación con Bradyrhizobium japonicum y Azospirillum brasilense con relación a la inoculación simple con Bradyrhizobium japonicum en la nodulación, crecimiento y acumulación de N en el cultivo de soja y su rendimiento. Se realizaron dos experimentos en lotes en los que nunca se cultivó soja en años con diferentes condiciones hídricas. Los tratamientos fueron: testigo (sin inoculación, coinoculación con Bradyrhizobium japonicum y Azospirillum brasilense e inoculación simple con Bradyrhizobium japonicum. La nodulación en ambos experimentos fue superior en la coinoculación, aunque hubo diferencias significativas en el experimento con déficit hídrico. Los rendimientos, en ambos experimentos, presentaron valores medios superiores en la coinoculación, aunque sin diferencias significativas. La acumulación de biomasa aérea y N fue mayor en el año húmedo. El porcentaje de N derivado de la fijación biológica de N en el cultivo en el año seco fue de 26% para la inoculación simple y de 30% en la coinoculación, mientras que en el año húmedo este porcentaje fue de 41 y 47% respectivamente. A partir de estos resultados se concluye que existe un efecto de estimulación del crecimiento del cultivo de soja por la coinoculación. Sin embargo, es necesario continuar con estudios sobre este tema.The aim of this study was to compare coinoculation with Bradyrhizobium japonicum and Azospirillum brasilense with inoculation with Bradyrhizobium japonicum on nodulation, growth, and N accumulation in the soybean crop in field conditions. Two field tests were carried out for two years with different hydric conditions in areas not previously cultivated with soybean. The treatments were: control, coinoculation using Bradyrhizobium japonicum + Azospirillum brasilense and inoculation with Bradyrhizobium japonicum. Nodulation in both experiments was increased by dual inoculation, although significant differences were

  14. Ultrastructural and immunocytochemical studies on the MLS-type basal apparatus in the sperm of Lygodium japonicum (pteridophyta)

    OpenAIRE

    Sakaushi, Shinji

    2004-01-01

    Spiral formation of the multiflagellated sperm in Lygodium japonicum(pteridophyte) was ultrastructurally and biochemically analyzed to consider evolution of the basal apparatus in the archegoniates.Differentiation of the spermatid (spermiogenesis) was investigated in detail by electron microscopy, immunofluorescence and immunoelectron microscopy using anti-tubulin and anti-actin antibodies. At the mid stage of spermiogenesis, the MLS, a characteristic structure in the archegoniates and the C...

  15. Practical application of methanol-mediated mutualistic symbiosis between Methylobacterium species and a roof greening moss, Racomitrium japonicum.

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    Akio Tani

    Full Text Available Bryophytes, or mosses, are considered the most maintenance-free materials for roof greening. Racomitrium species are most often used due to their high tolerance to desiccation. Because they grow slowly, a technology for forcing their growth is desired. We succeeded in the efficient production of R. japonicum in liquid culture. The structure of the microbial community is crucial to stabilize the culture. A culture-independent technique revealed that the cultures contain methylotrophic bacteria. Using yeast cells that fluoresce in the presence of methanol, methanol emission from the moss was confirmed, suggesting that it is an important carbon and energy source for the bacteria. We isolated Methylobacterium species from the liquid culture and studied their characteristics. The isolates were able to strongly promote the growth of some mosses including R. japonicum and seed plants, but the plant-microbe combination was important, since growth promotion was not uniform across species. One of the isolates, strain 22A, was cultivated with R. japonicum in liquid culture and in a field experiment, resulting in strong growth promotion. Mutualistic symbiosis can thus be utilized for industrial moss production.

  16. Genetic manipulation of Schistosoma haematobium, the neglected schistosome.

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    Gabriel Rinaldi

    2011-10-01

    Full Text Available BACKGROUND: Minimal information on the genome and proteome of Schistosoma haematobium is available, in marked contrast to the situation with the other major species of human schistosomes for which draft genome sequences have been reported. Accordingly, little is known about functional genomics in S. haematobium, including the utility or not of RNA interference techniques that, if available, promise to guide development of new interventions for schistosomiasis haematobia. METHODS/FINDINGS: Here we isolated and cultured developmental stages of S. haematobium, derived from experimentally infected hamsters. Targeting different developmental stages, we investigated the utility of soaking and/or square wave electroporation in order to transfect S. haematobium with nucleic acid reporters including Cy3-labeled small RNAs, messenger RNA encoding firefly luciferase, and short interfering RNAs (siRNAs. Three hours after incubation of S. haematobium eggs in 50 ng/µl Cy3-labeled siRNA, fluorescent foci were evident indicating that labeled siRNA had penetrated into miracidia developing within the egg shell. Firefly luciferase activity was detected three hours after square wave electroporation of the schistosome eggs and adult worms in 150 ng/µl of mRNA. RNA interference knockdown (silencing of reporter luciferase activity was seen following the introduction of dsRNA specific for luciferase mRNA in eggs, schistosomules and mixed sex adults. Moreover, introduction of an endogenous gene-specific siRNA into adult schistosomes silenced transcription of tetraspanin 2 (Sh-tsp-2, the apparent orthologue of the Schistosoma mansoni gene Sm-tsp-2 which encodes the surface localized structural and signaling protein Sm-TSP-2. Together, knockdown of reporter luciferase and Sh-tsp-2 indicated the presence of an intact RNAi pathway in S. haematobium. Also, we employed laser scanning confocal microscopy to view the adult stages of S. haematobium. CONCLUSIONS: These findings

  17. Schistosoma mansoni antigenic extracts obtained by different extraction procedures

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    Miriam Tendler

    1981-06-01

    Full Text Available Solubilization of Schistosoma mansoni antigens was obtained by agitation of adult worms in a 3M KCl solution. The protein contents of the KCl extrats varied from 0.35 to 0.96 mg/ml. Sera from 97 patients with hepatointestinal shistosomiasis and viable eggs in stools from a Brazilian endemic area were studied by immunoelectroomophoresis and Ouchterlony immunodiffusion methods with the KCl extract and with another antigen, obtained by homogenization of adult schistosomes in saline. The rate of positiveness of immunoprecipitation deterctions by immunoelectroomophoresis with the KCl extract was 53.5%. A correlation was verified between methods of detection and extration procedures, resulting in a better association of the extract obtained by agitation in 3M KCl and immunoelectroomophoresis.Foi obtida a solubilização de antígenos do Schistosoma mansoni por agitação de vermes adultos em solução de KCl 3M. O teor protéico dos extratos de KCl variou de 0,35 a 0,96mg/ ml. Foram testados pelos métodos de imunoeletroosmoforese (IEOP e dupla imunodifusão (Ouchterlony, 97 soros de doentes de area endêmica brasileira de esquistossomose, forma clínica hepatointestinal e com exames coprológicos positivos para S. mansoni, com o extrato de KCl e outro antígeno obtido pela homogenização de vermes adultos em salina. A taxa de positividade das reações de imunoprecipitação por IEOP com o antígeno extraído pela ação do KCl 3M foi 53,5%. Foi verificada a correlação entre os métodos de detecção e de extração resultando numa melhor associação entre o extrato obtido por agitação no KCl 3M e a IEOP.

  18. Recent advances in the diagnosis of Schistosoma infection: the detection of parasite DNA.

    Science.gov (United States)

    Rabello, Ana; Pontes, Luís André; Dias-Neto, Emmanuel

    2002-01-01

    As Schistosoma sp. control programs are chiefly based on treatment of infected population, adequate case finding has a crucial role. The available diagnostic methods are far from ideal, since the search for eggs in stools and the detection of circulating antigens lack sensitivity in low prevalence and post-treatment situations and antibody detection lacks specificity. In most endemic foci, repeated treatment of infected people leaves a number of non-diagnosed and consequently non-treated persons, enough to maintain a persistent residue of 5 to 10% prevalence. In an attempt to surpass these diagnostic limitations we have developed a polymerase chain reaction (PCR) for the detection of Schistosoma sp. in feces that, in a first population study, has shown to be more sensitive than three-repeated stool Kato-Katz examination. The PCR may constitute a valuable tool for the diagnosis of the Schistosoma sp. infection in special situations, when high sensitivity and specificity are required and infrastructure is available.

  19. Evolution of sarcoma 180 (ascitic tumor in mice infected with Schistosoma mansoni

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    Fausto Edmundo Lima Pereira

    1986-03-01

    Full Text Available Mice infected with 60 cercariae of Schistosoma mansoni were more resistant to the sarcoma 180 ascites tumor. Tumor inoculation was performed 50 days after schistosoma infection and the animals were observed and weighed at 48 hours intervals for development and progression of malignancy. In infected mice the weight gain (ascites formation started later and was shorter than in uninfected Controls. Also, the number of tumor cells into the peritoneal cavity 72h after tumor implantation was shorter in infected group than incontrols. This in creased resistance against a transplantable tumor probably is related to the effect of endotoxin on tumoricidal activity of macrophages activated by the infection. The immunodepression induced by Schistosoma mansoni infection enhances the proliferation of endogenous bacteria increasing the amount of endotoxin absorbed from the gut.

  20. Preliminary analysis of miRNA pathway in Schistosoma mansoni.

    Science.gov (United States)

    Gomes, Matheus S; Cabral, Fernanda J; Jannotti-Passos, Liana K; Carvalho, Omar; Rodrigues, Vanderlei; Baba, Elio H; Sá, Renata G

    2009-03-01

    RNA silencing refers to a series of nuclear and cytoplasmatic processes involved in the post-transcriptional regulation of gene expression or post-transcriptional gene silencing (PTGS), either by sequence-specific mRNA degradation or by translational arrest. The best characterized small RNAs are microRNAs (miRNAs), which predominantly perform gene silencing through post-transcriptional mechanisms. In this work we used bioinformatic approaches to identify the parasitic trematode Schistosoma mansoni sequences that are similar to enzymes involved in the post-transcriptional gene silencing mediated by miRNA pathway. We used amino acid sequences of well-known proteins involved in the miRNA pathway against S. mansoni genome and transcriptome databases identifying a total of 13 putative proteins in the parasite. In addition, the transcript levels of SmDicer1 and SmAgo2/3/4 were identified by qRT-PCR using cercariae, adult worms, eggs and in vitro cultivated schistosomula. Our results showed that the SmDicer1 and SmAgo2/3/4 are differentially expressed during schistosomula development, suggesting that the miRNA pathway is regulated at the transcript level and therefore may control gene expression during the life cycle of S. mansoni.

  1. Modeling and analysis of Schistosoma Argonaute protein molecular spatial conformation

    Institute of Scientific and Technical Information of China (English)

    Jianhua Zhang; Zhigang Shang; Xiaohui Zhang; Yuntao Zhang

    2011-01-01

    Objective: To analyze the amino acid sequence composition, secondary structure, the spatial conformation of its domain and other characteristics of Argonaute protein. Methods:Bioinformatics tools and the internet server were used. Firstly, the amino acid sequence composition features of the Argonaute protein were analyzed, and the phylogenetic tree was constructed. Secondly, Argonaute protein’s distribution of secondary structure and its physicochemical properties were predicted. Lastly, the protein functional expression form of the domain group was established through the Phyre-based analysis on the spatial conformation of Argonaute protein domains. Results: 593 amino acids were encoded by Argonaute protein, the phylogenetic tree was constructed, and Argonaute protein’s distribution of secondary structure and its physicochemical properties were obtained through analysis. In addition, the functional expression form which comprised the N-terminal PAZ domain and C-terminal Piwi domain for the Argonaute protein was obtained with Phyre. Conclusions: The information relationship between the structure and function of the Argonaute protein can be initially established with bioinformatics tools and the internet server, and this provides the theoretical basis for further clarifying the function of Schistosoma Argonaute protein.

  2. Compatibility of Schistosoma mansoni Cameroon and Biomphalaria pfeifferi Senegal.

    Science.gov (United States)

    Southgate, V R; Tchuenté, L A; Théron, A; Jourdane, J; Ly, A; Moncrieff, C B; Gryseels, B

    2000-11-01

    The vectorial capacity of Biomphalaria pfeifferi from Ndiangue, Senegal, was investigated with an allopatric isolate of Schistosoma mansoni from Nkolbisson, Cameroon. The snail infection rate after exposure to a single miracidium per snail (MD1) was 56. 3 %, and 91.6%, for snails exposed to 5 miracidia per snail (MD5). The minimum pre-patent period was 21 days. The mean total cercarial production for the MDI group was 18,511 cercariae per snail, and 9757 cercariae for the MD5 group. The maximum production of cercariae for 1 day was 4892 observed in a snail from the MDI group at day 43 post-infection. The mean longevity of snails was higher in group MD1 (88 days p.i.) than in group MD5 (65 days p.i.). The chronobiological emergence pattern revealed a circadian rhythm with one shedding peak at mid-day. Comparisons are made with the vectorial capacity of the sympatric combination of B. pfeifferi Senegal/S. mansoni Senegal.

  3. Uncovering Notch pathway in the parasitic flatworm Schistosoma mansoni.

    Science.gov (United States)

    Magalhães, Lizandra G; Morais, Enyara R; Machado, Carla B; Gomes, Matheus S; Cabral, Fernanda J; Souza, Julia M; Soares, Cláudia S; Sá, Renata G; Castro-Borges, William; Rodrigues, Vanderlei

    2016-10-01

    Several signaling molecules that govern development in higher animals have been identified in the parasite Schistosoma mansoni, including the transforming growth factor β, protein tyrosine kinases, nuclear hormone receptors, among others. The Notch pathway is a highly conserved signaling mechanism which is involved in a wide variety of developmental processes including embryogenesis and oogenesis in worms and flies. Here we aimed to provide the molecular reconstitution of the Notch pathway in S. mansoni using the available transcriptome and genome databases. Our results also revealed the presence of the transcripts coded for SmNotch, SmSu(H), SmHes, and the gamma-secretase complex (SmNicastrin, SmAph-1, and SmPen-2), throughout all the life stages analyzed. Besides, it was observed that the viability and separation of adult worm pairs were not affected by treatment with N-[N(3,5)-difluorophenacetyl)-L-Alanyl]-S-phenylglycine t-butyl ester (DAPT), a Notch pathway inhibitor. Moreover, DAPT treatment decreased the production of phenotypically normal eggs and arrested their development in culture. Our results also showed a significant decrease in SmHes transcript levels in both adult worms and eggs treated with DAPT. These results provide, for the first time, functional validation of the Notch pathway in S. mansoni and suggest its involvement in parasite oogenesis and embryogenesis. Given the complexity of the Notch pathway, further experiments shall highlight the full repertoire of Notch-mediated cellular processes throughout the S. mansoni life cycle.

  4. Identification of Schistosoma mansoni candidate antigens for diagnosis of schistosomiasis

    Directory of Open Access Journals (Sweden)

    Gardenia Braz Figueiredo Carvalho

    2011-11-01

    Full Text Available The development of a more sensitive diagnostic test for schistosomiasis is needed to overcome the limitations of the use of stool examination in low endemic areas. Using parasite antigens in enzyme linked immunosorbent assay is a promising strategy, however a more rational selection of parasite antigens is necessary. In this study we performed in silico analysis of the Schistosoma mansoni genome, using SchistoDB database and bioinformatic tools for screening immunogenic antigens. Based on evidence of expression in all parasite life stage within the definitive host, extracellular or plasmatic membrane localization, low similarity to human and other helminthic proteins and presence of predicted B cell epitopes, six candidates were selected: a glycosylphosphatidylinositol-anchored 200 kDa protein, two putative cytochrome oxidase subunits, two expressed proteins and one hypothetical protein. The recognition in unidimensional and bidimensional Western blot of protein with similar molecular weight and isoelectric point to the selected antigens by sera from S. mansoni infected mice indicate a good correlation between these two approaches in selecting immunogenic proteins.

  5. Serological screening of the Schistosoma mansoni adult worm proteome.

    Directory of Open Access Journals (Sweden)

    Fernanda Ludolf

    2014-03-01

    Full Text Available BACKGROUND: New interventions tools are a priority for schistosomiasis control and elimination, as the disease is still highly prevalent. The identification of proteins associated with active infection and protective immune response may constitute the basis for the development of a successful vaccine and could also indicate new diagnostic candidates. In this context, post-genomic technologies have been progressing, resulting in a more rational discovery of new biomarkers of resistance and antigens for diagnosis. METHODOLOGY/PRINCIPAL FINDINGS: Two-dimensional electrophoresed Schistosoma mansoni adult worm protein extracts were probed with pooled sera of infected and non-infected (naturally resistant individuals from a S. mansoni endemic area. A total of 47 different immunoreactive proteins were identified by mass spectrometry. Although the different pooled sera shared most of the immunoreactive protein spots, nine protein spots reacted exclusively with the serum pool of infected individuals, which correspond to annexin, major egg antigen, troponin T, filamin, disulphide-isomerase ER-60 precursor, actin and reticulocalbin. One protein spot, corresponding to eukaryotic translation elongation factor, reacted exclusively with the pooled sera of non-infected individuals living in the endemic area. Western blotting of two selected recombinant proteins, major egg antigen and hemoglobinase, showed a similar recognition pattern of that of the native protein. CONCLUDING/SIGNIFICANCE: Using a serological proteome analysis, a group of antigens related to the different infection status of the endemic area residents was identified and may be related to susceptibility or resistance to infection.

  6. Inheritance of Schistosoma mansoni infection incompatibility in Biomphalaria alexandrina snails

    Directory of Open Access Journals (Sweden)

    Iman F Abou El Naga

    2010-03-01

    Full Text Available In this study, we looked at the inheritance of susceptibility and resistance to Schistosoma mansoni infection in the first generation of crossbred Biomphalaria alexandrina snails. Our ultimate goal is to use such information to develop a biological method of controlling schistosomiasis. We infected laboratory-bred snails with S. mansoni miracidia and examined cercarial shedding to determine susceptibility and resistance. Five parental groups were used: Group I contained 30 susceptible snails, Group II contained 30 resistant snails, Group III contained 15 susceptible and 15 resistant snails, Group IV contained 27 susceptible and three resistant snails and Group V contained three susceptible and 27 resistant snails. The percentage of resistant snails in the resulting progeny varied according to the ratio of susceptible and resistant parents per group; they are 7%, 100%, 68%, 45% and 97% from Groups I, II, III, IV and V, respectively. On increasing the percentage of resistant parent snails, the percentage of resistant progeny increased, while cercarial production in their susceptible progeny decreased.

  7. The genome of the blood fluke Schistosoma mansoni

    Science.gov (United States)

    Berriman, Matthew; Haas, Brian J.; LoVerde, Philip T.; Wilson, R. Alan; Dillon, Gary P.; Cerqueira, Gustavo C.; Mashiyama, Susan T.; Al-Lazikani, Bissan; Andrade, Luiza F.; Ashton, Peter D.; Aslett, Martin A.; Bartholomeu, Daniella C.; Blandin, Gaelle; Caffrey, Conor R.; Coghlan, Avril; Coulson, Richard; Day, Tim A.; Delcher, Art; DeMarco, Ricardo; Djikeng, Appoliniare; Eyre, Tina; Gamble, John A.; Ghedin, Elodie; Gu, Yong; Hertz-Fowler, Christiane; Hirai, Hirohisha; Hirai, Yuriko; Houston, Robin; Ivens, Alasdair; Johnston, David A.; Lacerda, Daniela; Macedo, Camila D.; McVeigh, Paul; Ning, Zemin; Oliveira, Guilherme; Overington, John P.; Parkhill, Julian; Pertea, Mihaela; Pierce, Raymond J.; Protasio, Anna V.; Quail, Michael A.; Rajandream, Marie-Adèle; Rogers, Jane; Sajid, Mohammed; Salzberg, Steven L.; Stanke, Mario; Tivey, Adrian R.; White, Owen; Williams, David L.; Wortman, Jennifer; Wu, Wenjie; Zamanian, Mostafa; Zerlotini, Adhemar; Fraser-Liggett, Claire M.; Barrell, Barclay G.; El-Sayed, Najib M.

    2009-01-01

    Schistosoma mansoni is responsible for the neglected tropical disease schistosomiasis that affects 210 million people in 76 countries. We report here analysis of the 363 megabase nuclear genome of the blood fluke. It encodes at least 11,809 genes, with an unusual intron size distribution, and novel families of micro-exon genes that undergo frequent alternate splicing. As the first sequenced flatworm, and a representative of the lophotrochozoa, it offers insights into early events in the evolution of the animals, including the development of a body pattern with bilateral symmetry, and the development of tissues into organs. Our analysis has been informed by the need to find new drug targets. The deficits in lipid metabolism that make schistosomes dependent on the host are revealed, while the identification of membrane receptors, ion channels and more than 300 proteases, provide new insights into the biology of the life cycle and novel targets. Bioinformatics approaches have identified metabolic chokepoints while a chemogenomic screen has pinpointed schistosome proteins for which existing drugs may be active. The information generated provides an invaluable resource for the research community to develop much needed new control tools for the treatment and eradication of this important and neglected disease. PMID:19606141

  8. UVB-induced immune suppression and infection with Schistosoma mansoni

    Energy Technology Data Exchange (ETDEWEB)

    Noonan, F.P.; Lewis, F.A. [George Washington Univ., Washington, DC (United States). School of Medicine]|[Biomedical Research Inst., Rockville, MD (United States)

    1995-01-01

    Irradiation with ultraviolet B (UVB, 290-320 nm) causes a systematic immunosuppression of cell-mediated immunity. The question of whether UV immunosuppression modulates the course of infectious diseases is important because UVB levels in sunlight are sufficient to predict significant UV-induced immunosuppression at most latitudes. We have investigated the effect of immunosuppressive doses of UVB on the disease caused by the helminth parasite Schistosoma mansoni. C57BL/6 mice were irradiated once or three times weekly over 60-80 days with UV from a bank of FS40 sunlamps. Each UV treatment consisted of an immunosuppressive UV dose, as determined by suppression of contact hypersensitivity to trinitrochlorobenzene, corresponding to about 15-30 min of noonday tropical sunlight exposure under ideal clear sky conditions. Cumulative UV doses were between 80 and 170 kJ/m{sup 2}. Worm and egg burdens, liver granuloma diameters and liver fibrosis showed minimal changes (< 20%) compared with parameters in unirradiated animals. Ultraviolet irradiation (a total of 55 kJ/m{sup 2} administered in six treatments) did not impair the resistance to rechallenge conferred by vaccination with {sup 60}Co-irradiated cercariae. We have observed a dichotomy between UV immnosuppression and both disease and vaccination in this helminth infection, in contrast to the effects of UVB shown in other infectious diseases. (author).

  9. Behavior of Bradyrhizobium japonicum strains under different herbicide concentrations Comportamento de estirpes de Bradyrhizobium japonicum em diferentes concentrações de herbicidas

    Directory of Open Access Journals (Sweden)

    J.S. Arruda

    2001-04-01

    Full Text Available The Bradyrhizobium japonicun strains SEMIA 5073, SEMIA 5074, SEMIA 5079 and SEMIA 5080 were grown in vitro using Vincent medium combined with different rates of the herbicides imazaquin (0, 0.04, 0.12, 0.24, 0.36 mg a.i. g-1, clomazone (0, 0.4, 0.8, 1.6 and 3.2 mg a.i. g-1 and sulfentrazone (0, 0.2, 0.4, 0.8 and 1.6 mg a.i. g-1 to evaluate the strains tolerance to herbicides. The three herbicides drastically inhibited all the rhizobium strains tested, showing a significant decrease of the CFU number as a function of herbicide rates. The rhizobium strains presented a differentiated tolerance to the herbicides. The herbicide rates that reduced 50% (I50 of the growth or survival of the rhizobium strains were below the recommended sprayed rates for weed control in the soybean crop, for all the three herbicides studied; however, sulfentrazone I50 was smaller than imazaquin and clomazone I50.As estirpes de Bradyrhizobium japonicum SEMIA 5073, SEMIA 5074, SEMIA 5079 e SEMIA 5080 foram cultivadas in vitro, utilizando meio de Vincent acrescido de diferentes doses dos herbicidas imazaquin (0; 0,04; 0,12; 0,24; 0,36 mg i.a. g-1, clomazone (0; 0,4; 0,8; 1,6; 3,2 m i.a. g-1 e sulfentrazone (0; 0,2; 0,4; 0,8; 1,6 mg i.a. g-1, com o objetivo de avaliar a tolerância destas estirpes aos herbicidas. Os três herbicidas afetaram drasticamente as estirpes de rizóbio testadas, causando decréscimo significativo no crescimento ou na sobrevivência, em função do incremento nas doses. As estirpes de rizóbio apresentaram tolerância diferenciada aos herbicidas. As doses de herbicidas que reduziram em 50% o crescimento ou a sobrevivência das estirpes (I50 foram inferiores àquelas recomendadas para o controle de plantas daninhas na cultura de soja, para os três herbicidas estudados; no entanto, o I50 do herbicida sulfentrazone foi significativamente menor que o I50 dos herbicidas imazaquin e clomazone.

  10. The susceptibility of rodents to schistosome infection, with special reference to Schistosoma haematobium.

    Science.gov (United States)

    Gear, J H; Davis, D H; Pitchford, R J

    1966-01-01

    In this investigation the susceptibility of several species of rodents-Praomys (Mastomys) natalensis, Saccostomus campestris, Arvicanthus niloticus, Aethomys chrysophilus, Tatera brantsi and the white mouse (SAIMR 200 strain)-to Schistosoma haematobium was determined and the pathology studied. From the results it is clear that these rodents are susceptible to infection with Schistosoma haematobium. For various reasons, notably adaptability to laboratory conditions, the most suitable as laboratory animals for the study of bilharziasis are Saccostomus campestris, Arvicanthus niloticus and Praomys (Mastomys) natalensis. These three species breed readily in the laboratory and show a high susceptibility to S. haematobium, with characteristic lesions involving several organs, including the lungs, liver, spleen, pancreas and intestine.

  11. Ultrasound guided supraclavicular block.

    LENUS (Irish Health Repository)

    Hanumanthaiah, Deepak

    2013-09-01

    Ultrasound guided regional anaesthesia is becoming increasingly popular. The supraclavicular block has been transformed by ultrasound guidance into a potentially safe superficial block. We reviewed the techniques of performing supraclavicular block with special focus on ultrasound guidance.

  12. A multistrain approach to studying the mechanisms underlying compatibility in the interaction between Biomphalaria glabrata and Schistosoma mansoni.

    Science.gov (United States)

    Galinier, Richard; Roger, Emmanuel; Moné, Yves; Duval, David; Portet, Anaïs; Pinaud, Silvain; Chaparro, Cristian; Grunau, Christoph; Genthon, Clémence; Dubois, Emeric; Rognon, Anne; Arancibia, Nathalie; Dejean, Bernard; Théron, André; Gourbal, Benjamin; Mitta, Guillaume

    2017-03-01

    In recent decades, numerous studies have sought to better understand the mechanisms underlying the compatibility between Biomphalaria glabrata and Schistosoma mansoni. The developments of comparative transcriptomics, comparative genomics, interactomics and more targeted approaches have enabled researchers to identify a series of candidate genes. However, no molecular comparative work has yet been performed on multiple populations displaying different levels of compatibility. Here, we seek to fill this gap in the literature. We focused on B. glabrata FREPs and S. mansoni SmPoMucs, which were previously demonstrated to be involved in snail/schistosome compatibility. We studied the expression and polymorphisms of these factors in combinations of snail and schistosome isolates that display different levels of compatibility. We found that the polymorphism and expression levels of FREPs and SmPoMucs could be linked to the compatibility level of S. mansoni. These data and our complementary results obtained by RNA-seq of samples from various snail strains indicate that the mechanism of compatibility is much more complex than previously thought, and that it is likely to be highly variable within and between populations. This complexity must be taken into account if we hope to identify the molecular pathways that are most likely to be good targets for strategies aimed at blocking transmission of the parasite through the snail intermediate host.

  13. Development of oral and branchial muscles in lancelet larvae of Branchiostoma japonicum.

    Science.gov (United States)

    Yasui, Kinya; Kaji, Takao; Morov, Arseniy R; Yonemura, Shigenobu

    2014-04-01

    The perforated pharynx has generally been regarded as a shared characteristic of chordates. However, there still remains phylogenetic ambiguity between the cilia-driven system in invertebrate chordates and the muscle-driven system in vertebrates. Giant larvae of the genus Asymmetron were reported to develop an orobranchial musculature similar to that of vertebrates more than 100 years ago. This discovery might represent an evolutionary link for the chordate branchial system, but few investigations of the lancelet orobranchial musculature have been completed since. We studied staged larvae of a Japanese population of Branchiostoma japonicum to characterize the developmental property of the orobranchial musculature. The larval mouth and the unpaired primary gills develop well-organized muscles. These muscles function only as obturators of the openings without antagonistic system. As the larval mouth enlarged posteriorly to the level of the ninth myomere, the oral musculature was fortified accordingly without segmental patterning. In contrast, the iterated branchial muscles coincided with the dorsal myomeric pattern before metamorphosis, but the pharynx was remodeled dynamically irrespective of the myomeric pattern during metamorphosis. The orobranchial musculature disappeared completely during metamorphosis, and adult muscles in the oral hood and velum, as well as on the pterygial coeloms developed independently. The lancelet orobranchial musculature is apparently a larval adaptation to prevent harmful intake. However, vestigial muscles appeared transiently with the secondary gill formation suggest a bilateral ancestral state of muscular gills, and a segmental pattern of developing branchial muscles without neural crest and placodal contributions is suggestive of a precursor of vertebrate branchiomeric pattern.

  14. EFFECT OF FERULIC ACID ON CHEMOTAXIS AND NODULATION OF Bradyrhizobium japonicum

    Directory of Open Access Journals (Sweden)

    María C. Nápoles

    2001-01-01

    Full Text Available En este trabajo se estudió el efecto de tres concentraciones de ácido ferúlico sobre la quimiotaxis y la nodulación de B. japonicum ICA 8001. También se evaluó el efecto de este ácido hidroxicinámico obtenido a partir de vainillina, así como su capacidad de inducción sobre los genes de nodulación mediante la detección de factores Nod sintetizados. Se obtuvo una actividad quimiotáctica positiva pero no fuerte y solamente 10 mM como componente del medio de cultivo mostró una influencia positiva sobre la nodulación. El ácido ferúlico sintetizado a partir de vainillina incrementó todos los parámetros de la nodulación. La actividad nod inductora de este ácido se evidenció con la producción de cuatro estructuras de lipoquitinoligosacáridos.

  15. The major Nod factor of Bradyrhizobium japonicum promotes early growth of soybean and corn.

    Science.gov (United States)

    Souleimanov, A; Prithiviraj, B; Smith, D L

    2002-09-01

    Greenhouse experiments were conducted to evaluate the effect of Nod factor Nod Bj-V (C18:1, MeFuc) of Badyrhizobium japonicum on the growth of soybean and corn. Three-day-old seedlings of soybean and corn were grown in hydroponic solutions containing four concentrations (0, 10(-7), 10(-9) or 10(-11) M) of Nod factor. After 7 d of treatment, Nod factor enhanced soybean and corn biomass. Nod factor elicited profound effects on root growth resulting in 34-44% longer roots in soybean. More detailed analyses of the roots, using a scanner based image analysis system, revealed that Nod factor increased the total length, projected area and surface area of the roots and decreased the diameter of soybean roots, while it increased the total length of corn roots. Stem injection of soybean plants with 10(-7) M Nod factor resulted in increased dry matter accumulation. These results suggest that Nod factor, besides mediating early stages of nodulation, has more general plant growth-promoting effects.

  16. Proteomic Analysis of Soybean [Glycine max (L.) Merrill] Roots Inoculated with Bradyrhizobium japonicum Strain CPAC 15.

    Science.gov (United States)

    Torres, Adalgisa R; Rodrigues, Elisete P; Batista, Jesiane Ss; Gomes, Douglas F; Hungria, Mariangela

    2013-01-01

    This research intended to analyze the expression pattern of proteins in roots of the Brazilian soybean cultivar Conquista when inoculated with Bradyrhizobium japonicum CPAC 15, a strain broadly used in commercial inoculants in Brazil. At ten days after bacterial inoculation, whole-cell proteins were extracted from roots and separated by 2-D gel electrophoresis. Comparative analysis revealed significant changes in the intensity of 37 spots due to the inoculation (17 up-regulated and 20 down-regulated proteins), identified by MALDI-TOF/TOF-TOF. Identified proteins were associated with COG functional categories of information storage and processing, cellular processes and signaling, metabolism, and also in the "poorly characterized" and "not in COG" categories. Among the up-regulated proteins, we identified sucrose synthase (nodulin-100), β-tubulin, rubisco activase, glutathione-S-transferase, a putative heat-shock 70-kDa protein, pyridine nucleotide-disulphideoxidoreductase and a putative transposase. Proteomic analysis allowed for the identification of some putative symbiotic functions and confirmed the main biological processes triggered in the nitrogen-fixing symbiosis with soybean.

  17. Molluscicidal Activity of Nerium indicum Mill, Pterocarya stenoptera DC, and Rumex japonicum Houtt on Oncomelania hupensis

    Institute of Scientific and Technical Information of China (English)

    HONG WANG; WEI-MIN CAI; WAN-XIAN WANG; JIAN-MIN YANG

    2006-01-01

    To evaluate the molluscicidal activities of three Chinese plants N. Indicum Mill, P. Stenoptera DC, and R.japonicum Houtt, and to clarify the molluscicidal mechanism. Methods N-butanol extracts and water extracts of the three plants were obtained. The reactions of EST isozyme, glycogen and total protein of snails to the plant extracts were studied.Results EST electrophoresis showed that EST was an important antidotal enzyme system and reacted strongly to environment.EST changed greatly during the whole exposure period so that it could be viewed as a pathological index of toxicity. Extracts decreased the glycogen content of the snails'soft tissues greatly, and also the protein content. Conclusion All extracts show strong molluscicidal activity. The LD50 value of the water extract of N. Indicum Mill is as low as 13.2 mg/L. EST can be viewed as a pathological index of toxicity. The energy metabolism abnormity is the key reason for the molluscicidal activities. The biochemical mechanism needs further research.

  18. Caracterização de cepas de Schistosoma mansoni por morfometria de vermes adultos provenientes de infecção unissexual Characterization of Schistosoma mansoni strains by morphometry of adult worms derived from single-sex infection

    Directory of Open Access Journals (Sweden)

    José Roberto Machado-Silva

    2003-12-01

    Full Text Available Camundongos foram infectados com cercárias, de um único sexo, de cepas simpátricas do Schistosoma mansoni. Nos vermes adultos, foram encontradas diferenças significativas (pMice were infected with only one sex cercaria derived from sympatric strains of Schistosoma mansoni. Adult worms presented significative differences (p<0.05 regarding suckers, testicular lobes, ovary and thickness of the tegument. Data show that morphometric study of unisexual infection worms can be also used for characterization of Schistosoma mansoni strains.

  19. Simple school questionnaires can map both Schistosoma mansoni and Schistosoma haematobium in the Democratic Republic of Congo.

    Science.gov (United States)

    Lengeler, C; Makwala, J; Ngimbi, D; Utzinger, J

    2000-01-05

    The use of self-administered questionnaires has been shown in different African countries to be inexpensive and reliable for the rapid identification of communities at highest risk of urinary schistosomiasis. For intestinal schistosomiasis due to Schistosoma mansoni there is a clear need for a similar approach. We report the results from a large-scale study undertaken in the western part of the Democratic Republic of Congo (DRC, formerly Zaire). Within 4 weeks questionnaires were correctly completed in 136 out of 160 schools (85%). In 57 of these schools children were screened for infections with schistosomes and geohelminths. The prevalence of 'schistosomiasis' as reported in the questionnaires showed the best correlation with the prevalence of S. mansoni infections (r = 0.77, P schools with a high risk of intestinal schistosomiasis gave positive predictive values of 87 and 62%, and negative predictive values of 74 and 87% for moderate and high infection thresholds, respectively. Reported 'blood in stool' was another useful indicator for intestinal schistosomiasis. Reported 'blood in urine' showed the best correlation with urinary schistosomiasis (r = 0.75, P school children in DRC have a distinct perception of intestinal and urinary schistosomiasis and that questionnaires could be useful to identify high-risk schools for both parasites.

  20. A single-strand conformation polymorphism (SSCP) approach for investigating genetic interactions of Schistosoma haematobium and Schistosoma guineensis in Loum, Cameroon.

    Science.gov (United States)

    Webster, B L; Tchuem Tchuenté, L A; Southgate, V R

    2007-03-01

    Single-strand conformation polymorphism (SSCP) analysis of the second internal transcribed spacer (ITS2) of nuclear ribosomal DNA provides a molecular tool for the identification of Schistosoma haematobium, Schistosoma guineensis and the hybrids of these two species. This molecular tool was utilized to provide a detailed analysis of the interactions between S. haematobium and S. guineensis in hybrid zones of Loum, Littoral Province, Cameroon. Individual hybrid schistosomes were identified within the natural populations collected from Loum in 1990, 1999 and 2000, which would have been misidentified as S. haematobium using solely morphological and sequence criteria. This study indicates the complexities of the hybridization between S. haematobium and S. guineensis and emphasizes the importance of assessing morphological, biological and molecular data to gain insights into the interaction of these two species over time.

  1. Block Cipher Analysis

    DEFF Research Database (Denmark)

    Miolane, Charlotte Vikkelsø

    Block ciphersarecryptographicprimitivesthatoperateon fixed sizetexts(blocks). Mostdesigns aim towards secure andfastencryption oflarge amounts ofdata. Block ciphers also serve as the building block of a number of hash functions and message authentication codes(MAC).Thetask of cryptanalysisisto en...... on small scale variants of AES. In the final part of the thesis we present a new block cipher proposal Present and examine its security against algebraic and differential cryptanalysis in particular.......Block ciphersarecryptographicprimitivesthatoperateon fixed sizetexts(blocks). Mostdesigns aim towards secure andfastencryption oflarge amounts ofdata. Block ciphers also serve as the building block of a number of hash functions and message authentication codes(MAC).Thetask of cryptanalysisisto...... ensurethat no attack violatesthe securitybounds specifiedbygeneric attack namely exhaustivekey search and table lookup attacks. This thesis contains a general introduction to cryptography with focus on block ciphers and important block cipher designs, in particular the Advanced Encryption Standard...

  2. Interspecies Complementation of Escherichia coli ccm Mutants: CcmE (CycJ) from Bradyrhizobium japonicum Acts as a Heme Chaperone during Cytochrome c Maturation

    OpenAIRE

    Schulz, Henk; Thöny-Meyer, Linda

    2000-01-01

    Biogenesis of c-type cytochromes in α- and γ-proteobacteria requires the function of a set of orthologous genes (ccm genes) that encode specific maturation factors. The Escherichia coli CcmE protein is a periplasmic heme chaperone. The membrane protein CcmC is required for loading CcmE with heme. By expressing CcmE (CycJ) from Bradyrhizobium japonicum in E. coli we demonstrated that heme is bound covalently to this protein at a strictly conserved histidine residue. The B. japonicum homologue ...

  3. Characterization of the phytochelatin synthase of Schistosoma mansoni.

    Directory of Open Access Journals (Sweden)

    Debalina Ray

    2011-05-01

    Full Text Available Treatment for schistosomiasis, which is responsible for more than 280,000 deaths annually, depends exclusively on the use of praziquantel. Millions of people are treated annually with praziquantel and drug resistant parasites are likely to evolve. In order to identify novel drug targets the Schistosoma mansoni sequence databases were queried for proteins involved in glutathione metabolism. One potential target identified was phytochelatin synthase (PCS. Phytochelatins are oligopeptides synthesized enzymatically from glutathione by PCS that sequester toxic heavy metals in many organisms. However, humans do not have a PCS gene and do not synthesize phytochelatins. In this study we have characterized the PCS of S. mansoni (SmPCS. The conserved catalytic triad of cysteine-histidine-aspartate found in PCS proteins and cysteine proteases is also found in SmPCS, as are several cysteine residues thought to be involved in heavy metal binding and enzyme activation. The SmPCS open reading frame is considerably extended at both the N- and C-termini compared to PCS from other organisms. Multiple PCS transcripts are produced from the single encoded gene by alternative splicing, resulting in both mitochondrial and cytoplasmic protein variants. Expression of SmPCS in yeast increased cadmium tolerance from less than 50 µM to more than 1,000 µM. We confirmed the function of SmPCS by identifying PCs in yeast cell extracts using HPLC-mass spectrometry. SmPCS was found to be expressed in all mammalian stages of worm development investigated. Increases in SmPCS expression were seen in ex vivo worms cultured in the presence of iron, copper, cadmium, or zinc. Collectively, these results indicate that SmPCS plays an important role in schistosome response to heavy metals and that PCS is a potential drug target for schistosomiasis treatment. This is the first characterization of a PCS from a parasitic organism.

  4. Praziquantel treatment decreases Schistosoma mansoni genetic diversity in experimental infections.

    Directory of Open Access Journals (Sweden)

    Regina Coeli

    Full Text Available BACKGROUND: Schistosomiasis has a considerable impact on public health in many tropical and subtropical areas. In the new world, schistosomiasis is caused by the digenetic trematode Schistosoma mansoni. Chemotherapy is the main measure for controlling schistosomiasis, and the current drug of choice for treatment is praziquantel (PZQ. Although PZQ is efficient and safe, its repetitive large-scale use in endemic areas may lead to the selection of resistant strains. Isolates less susceptible to PZQ have been found in the field and selected for in the laboratory. The impact of selecting strains with a decreased susceptibility phenotype on disease dynamics and parasite population genetics is not fully understood. This study addresses the impact of PZQ pressure on the genetics of a laboratory population by analyzing frequency variations of polymorphic genetic markers. METHODOLOGY: Infected mice were treated with increasing PZQ doses until the highest dose of 3 × 300 mg/Kg was reached. The effect of PZQ treatment on the parasite population was assessed using five polymorphic microsatellite markers. Parasitological and genetic data were compared with those of the untreated control. After six parasite generations submitted to treatment, it was possible to obtain a S. mansoni population with decreased susceptibility to PZQ. In our experiments we also observed that female worms were more susceptible to PZQ than male worms. CONCLUSIONS: The selective pressure exerted by PZQ led to decreased genetic variability in S. mansoni and increased endogamy. The understanding of how S. mansoni populations respond to successive drug pressure has important implications on the appearance and maintenance of a PZQ resistance phenotype in endemic regions.

  5. Impact of natural and anthropogenic factors on the trophic interactions of molluscivores and Schistosoma host snails

    NARCIS (Netherlands)

    Monde, C.

    2016-01-01

    Summary With a global disease burden of 240 million infected people, of which 90% live in sub-Saharan Africa, schistosomiasis is one of the most prevalent Neglected Tropical Diseases affecting people in the tropical and sub-tropical regions of the world. Schistosoma parasites are th

  6. Partial molecular characterization of Sm8, a tegumental antigen of Schistosoma mansoni

    Directory of Open Access Journals (Sweden)

    Frederico GC Abath

    2002-10-01

    Full Text Available Sm8 is a major tegumental antigen of Schistosoma mansoni. The partial cDNA was isolated and analyzed. Sequence analysis revealed transmembrane compatible hydrophobic domains and a putative leucine zipper pattern. The mRNA and the protein are predominantly expressed in adult worms.

  7. Schistosoma mansoni antigens alter activation markers and cytokine profile in lymphocytes of patients with asthma.

    Science.gov (United States)

    de Almeida, Tarcísio Vila Verde Santana; Fernandes, Jamille Souza; Lopes, Diego Mota; Andrade, Lorena Santana; Oliveira, Sérgio Costa; Carvalho, Edgar M; Araujo, Maria Ilma; Cruz, Álvaro A; Cardoso, Luciana Santos

    2017-02-01

    Asthma is a chronic disease characterized by airway inflammation, obstruction and hyperresponsiveness. Severe asthma affects a small proportion of subjects but results in most of the morbidity, costs and mortality associated with the disease. Studies have suggested that Schistosoma mansoni infection reduces the severity of asthma and prevent atopy.

  8. Update of the Gene Discovery Program in Schistosoma mansoni with the Expressed Sequence Tag Approach

    Directory of Open Access Journals (Sweden)

    Élida ML Rabelo

    1997-09-01

    Full Text Available Continuing the Schistosoma mansoni Genome Project 363 new templates were sequenced generating 205 more ESTs corresponding to 91 genes. Seventy four of these genes (81% had not previously been described in S. mansoni. Among the newly discovered genes there are several of significant biological interest such as synaptophysin, NIFs-like and rho-GDP dissociation inhibitor

  9. Glycogen metabolism in Schistosoma mansoni worms after their isolation from the host

    NARCIS (Netherlands)

    Tiolens, A.G.M.; Bergh, S.G. van den

    1987-01-01

    Adult Schistosoma mansoni worms rapidly degrade their endogenous glycogen stores immediately after isolation from the host. In NCTC 109 or in a diphasic culture medium the glycogen levels slowly recovered again after the initial decrease. The rapid degradation of glycogen could be prevented, even in

  10. Increased prevalence of leukocytes and elevated cytokine levels in semen from Schistosoma haematobium-infected individuals

    DEFF Research Database (Denmark)

    Leutscher, Peter D C; Pedersen, Mette; Raharisolo, Clairette

    2005-01-01

    In this study, we investigated the seminal inflammatory response to egg infestation of the urogenital organs in 240 semen-donating men aged 15-49 years living in a Schistosoma haematobium-endemic area of Madagascar. In 29 subjects (12%) with excretion of > or =5 ova/ejaculate, leukocytospermia (>...

  11. Effects of Endosulfan on Predator–Prey Interactions Between Catfish and Schistosoma Host Snails

    NARCIS (Netherlands)

    Monde, Concillia; Syampungani, Stephen; Brink, van den Paul J.

    2016-01-01

    The effect of the pesticide endosulfan on predator–prey interactions between catfish and Schistosoma host snails was assessed in static tank experiments. Hybrid catfish (Clarias gariepinus × C. ngamensis) and Bulinus globosus were subjected to various endosulfan concentrations including an untrea

  12. The effect of praziquantel against Schistosoma mansoni-infections in Botswana

    DEFF Research Database (Denmark)

    Friis, H; Byskov, Jens

    1989-01-01

    Chemotherapy of all infected individuals, using praziquantel 40 mg/kg in a single dose, was the initial component of the recently introduced control programme against Schistosoma mansoni-infections in Ngamiland, Botswana. To evaluate the effect of praziquantel in Ngamiland, 81 children were...

  13. Binding of von Willebrand factor and plasma proteins to the eggshell of Schistosoma mansoni

    NARCIS (Netherlands)

    Dewalick, Saskia; Hensbergen, Paul J; Bexkens, Michiel L; Grosserichter-Wagener, Christina; Hokke, Cornelis H; Deelder, André M; de Groot, Philip G; Tielens, Aloysius G M; van Hellemond, Jaap J

    2014-01-01

    Schistosoma mansoni eggs have to cross the endothelium and intestinal wall to leave the host and continue the life cycle. Mechanisms involved in this essential step are largely unknown. Here we describe direct binding to the S. mansoni eggshell of von Willebrand factor and other plasma proteins invo

  14. C-type lectin interactions with Schistosoma mansoni SEA : Molecular basis and function

    NARCIS (Netherlands)

    Liempt, van P.A.G.

    2007-01-01

    Outline of this thesis The studies described in this thesis have been performed to gain more insight in the recognition of Schistosoma mansoni glycans by C-type lectins and the consequences for dendritic cell mediated immune responses. As a first approach to understand the molecular interactions o

  15. Imatinib treatment causes substantial transcriptional changes in adult Schistosoma mansoni in vitro exhibiting pleiotropic effects.

    Directory of Open Access Journals (Sweden)

    Christin Buro

    2014-06-01

    Full Text Available BACKGROUND: Schistosome parasites cause schistosomiasis, one of the most important infectious diseases worldwide. For decades Praziquantel (PZQ is the only drug widely used for controlling schistosomiasis. The absence of a vaccine and fear of PZQ resistance have motivated the search for alternatives. Studies on protein kinases (PKs demonstrated their importance for diverse physiological processes in schistosomes. Among others two Abl tyrosine kinases, SmAbl1 and SmAbl2, were identified in Schistosoma mansoni and shown to be transcribed in the gonads and the gastrodermis. SmAbl1 activity was blocked by Imatinib, a known Abl-TK inhibitor used in human cancer therapy (Gleevec/Glivec. Imatinib exhibited dramatic effects on the morphology and physiology of adult schistosomes in vitro causing the death of the parasites. METHODOLOGY/PRINCIPAL FINDINGS: Here we show modeling data supporting the targeting of SmAbl1/2 by Imatinib. A biochemical assay confirmed that SmAbl2 activity is also inhibited by Imatinib. Microarray analyses and qRT-PCR experiments were done to unravel transcriptional processes influenced by Imatinib in adult schistosomes in vitro demonstrating a wide influence on worm physiology. Surface-, muscle-, gut and gonad-associated processes were affected as evidenced by the differential transcription of e.g. the gynecophoral canal protein gene GCP, paramyosin, titin, hemoglobinase, and cathepsins. Furthermore, transcript levels of VAL-7 and egg formation-associated genes such as tyrosinase 1, p14, and fs800-like were affected as well as those of signaling genes including a ribosomal protein S6 kinase and a glutamate receptor. Finally, a comparative in silico analysis of the obtained microarray data sets and previous data analyzing the effect of a TGFβR1 inhibitor on transcription provided first evidence for an association of TGFβ and Abl kinase signaling. Among others GCP and egg formation-associated genes were identified as common

  16. An L-glucitol oxidizing dehydrogenase from Bradyrhizobium japonicum USDA 110 for production of D-sorbose with enzymatic or electrochemical cofactor regeneration

    DEFF Research Database (Denmark)

    Gauer, Sabrina; Wang, Zhijie; Otten, Harm;

    2014-01-01

    A gene in Bradyrhizobium japonicum USDA 110, annotated as a ribitol dehydrogenase (RDH), had 87 % sequence identity (97 % positives) to the N-terminal 31 amino acids of an L-glucitol dehydrogenase from Stenotrophomonas maltophilia DSMZ 14322. The 729-bp long RDH gene coded for a protein consistin...

  17. Plastome sequences of Lygodium japonicum and Marsilea crenata reveal the genome organization transformation from basal ferns to core leptosporangiates.

    Science.gov (United States)

    Gao, Lei; Wang, Bo; Wang, Zhi-Wei; Zhou, Yuan; Su, Ying-Juan; Wang, Ting

    2013-01-01

    Previous studies have shown that core leptosporangiates, the most species-rich group of extant ferns (monilophytes), have a distinct plastid genome (plastome) organization pattern from basal fern lineages. However, the details of genome structure transformation from ancestral ferns to core leptosporangiates remain unclear because of limited plastome data available. Here, we have determined the complete chloroplast genome sequences of Lygodium japonicum (Lygodiaceae), a member of schizaeoid ferns (Schizaeales), and Marsilea crenata (Marsileaceae), a representative of heterosporous ferns (Salviniales). The two species represent the sister and the basal lineages of core leptosporangiates, respectively, for which the plastome sequences are currently unavailable. Comparative genomic analysis of all sequenced fern plastomes reveals that the gene order of L. japonicum plastome occupies an intermediate position between that of basal ferns and core leptosporangiates. The two exons of the fern ndhB gene have a unique pattern of intragenic copy number variances. Specifically, the substitution rate heterogeneity between the two exons is congruent with their copy number changes, confirming the constraint role that inverted repeats may play on the substitution rate of chloroplast gene sequences.

  18. Cloning, Sequencing and Characterization of 3-Hydroxybuty- rate Dehydrogenase Encoding Gene (bdh A) in Bradyrhizobium japonicum USDA110 Strain

    Institute of Scientific and Technical Information of China (English)

    DAI Mei-xue; WU Bo; BAI Xue-liang; ZHANG Cheng-gang; MA Qing-sheng; Charles Trevor C

    2002-01-01

    The current study describes the molecular characterization of a clone which can restore the ability of bdhA mutant strains NGRPA2 and Rm11107 to utilize 3-hydroxybutyrate as a sole carbon source (Hbu+). This clone was screened out by complementation experiment from Bradyrhizobium japonicum USDA110 genomic library, and the presence of bdhA gene in the clone was verified by Bdh assay and Southern blot analysis. Furthermore, the entire sequence of bdhA gene was sequenced and the sequence was deposited in GenBank database under the accession number AY077581. bdhA gene comprises 789 base pairs and encodes Bdh with 262 amino acid of MW 27.59 kDa. Interposon ΩKm was inserted into the bdh A ORF at EcoR I site and the bdhA mutant was constructed in B .japonicum by homologous recombination. Plant assay result did not show obvious effects of mutation of bdhA gene on nodulation and nitrogen-fixation.

  19. Culturable bacteria in hydroponic cultures of moss Racomitrium japonicum and their potential as biofertilizers for moss production.

    Science.gov (United States)

    Tani, Akio; Akita, Motomu; Murase, Haruhiko; Kimbara, Kazuhide

    2011-07-01

    The use of Racomitrium japonicum, a drought resistant bryophyte used for roof-greening, is gradually increasing. However, its utilization is hampered by slow growth rate. Here we isolated culturable bacteria from hydroponic cultivation samples to identify isolates that could promote moss growth. Most of the isolates belonged to Pseudomonas, Rhodococcus, and Duganella species. The isolates were biochemically characterized according to their type of interaction with plants, i.e., production of auxin, siderophores, or hydrogen cyanate, growth in the absence of an added nitrogen source, calcium phosphate solubilization, utilization of sugars, polymers, or aliphatic compounds, and antifungal activity. The isolates were applied to sterile protonemata and non-sterile adult gametophytes of R. japonicum to evaluate their effect on plant growth. Furthermore, we isolated fungi that inhibited moss growth. Our results suggest that the microbial community structure in hydroponic cultures is important to stabilize moss production and the isolates that promote moss growth have potential to be utilized as biofertilizers for moss production.

  20. Immunization against Egyptian Schistosoma mansoni infection by multivalent DNA vaccine

    Institute of Scientific and Technical Information of China (English)

    Mahmoud H Romeih; Hanem M Hassan; Tarek S Abou Shousha; Mohamed A Saber

    2008-01-01

    The development of multivalent vaccines consisting of several antigens is a novel approach to creating broad-range protection against different parasite strains and parasite life cycle stages. We have previously confirmed that the schistosome Sm21.7 and SmFimbrin (SmFim) proteins could induce protection in mice. Therefore, this study aimed to construct the multivalent DNA vaccine Sm21.7-SmFim/pBudCE4.1 and evaluate its immune efficacy. The open reading frames of two Schistosoma mansoni genes, Sm21.7 and SmFim, were inserted into the eukaryotic expression plasmid pBudCE4.1 designed for the independent expression of two genes in mammalian cells. To evaluate the in vitro expression of the multivalent Sm21.7-SmFim/pBudCE4.1 DNA vaccine and its immunological effect in mice, the recombinant plasmid Sm21.7-SmFim/pBudCE4.1 was used to transfect 293T cells, and the expression of mRNA and proteins was examined using reverse transcription-polymerase chain reaction and Western blot analysis. Then the ability of Sm21.7.SmFim/pBudCE4.1 to protect against S. mansoni challenge infections was analyzed according to worm burden and egg reduction rates after vaccination of mice. Vaccinated mice showed a significant level of protection (56%), and a decrease in the number and size, and change in the cellular profile, of granulomas. Egg reduction in liver and intestine was 41.53% and 55.63%,respectively, as determined relative to mice that received the empty vector only. In addition to reductions in worm viability,worm fecundity and egg hatching ability were observed following challenge infection in the immunized group.Results showed that Sm21.7-SmFim/pBudCE4.1 could express Sm 21.7 and SmFim mRN A and proteins. Enzyme-linked immunosorbent assay and Western blot analysis indicated that immunized mice generated specific immunoglobulin G against Sm21.7-SmFim/pBudCE4.1. These results suggest that vaccination with multivalent S. mansoni DNA vaccine (SmFim-Sm21.7/pBudCE4.1) not only induces a

  1. Cloning and Expression of Metalloprotease Gene from Schistosoma japoncum and its Immunoprotective Efficiency%日本血吸虫金属蛋白酶基因的克隆和表达及其对小鼠的免疫保护性研究

    Institute of Scientific and Technical Information of China (English)

    徐斌; 鞠川; 卢艳; 莫筱谨; 冯正; 许学年; 胡薇

    2011-01-01

    Objective To clone and express a metalloprotease gene of Schistosoma japonicum, purify the expressed protein, and investigate the induced immune response in mice and its localization in the parasite. Methods Specific primers were designed according to the EST sequence and used for amplification of the encoding sequence from the S.japonicum cDNA clone containing S.japonicum metalloprotease. The gene was subcloned into pET-28a plasmid and expressed, and the recombinant protein was purified with HisoTag affinity chromatography. Western blotting was used to analyze the immunogenicity. Eighteen C57BC/6 mice were divided into two groups. Mice in group A were immunized each with 25 μg purified recombinant SjB04 at every 2 weeks for 3 times. Mice in group B received only adjuvant as control. Each mouse was challenged by (40±2) cercariae at the third week after the last immunization. Fecal samples were collected for 6 days from 37th days after challenge. Eggs per gram feces and rate of egg reduction were calculated. S.japonicum adult worms were collected from infected mice, and used for preparing frozen sections and indirect immunofluorescence staining with specific polyclone antibody to S. japonicum metalloprotease. Results The metalloprotease gene SjB04 was cloned, sequenced and expressed. The immuno-fluorescence localization showed that SjB04 protein distributed mainly in the intestinal epithelium of the adult worm. The recombinant protein was specifically recognized by the S. japonicum-infected rabbit sera, showing that the expressed product possessed antigenicity. Mice immunized with the recombinant protein revealed a reduction in numher of adult worms, eggs in feces by 27.1% and 57.8%,respectively. Conclusion The recombinant protein of' S.japonicum metalloprotease has been obtained with Mr 36500.The protein locates in the intestinal epithelium of adult worm. Immunization with the SjB04 protein induces significant reduction of fecal eggs.%目的 克隆和表达

  2. A new rapid diagnostic test for detection of anti-Schistosoma mansoni and anti-Schistosoma haematobium antibodies

    Directory of Open Access Journals (Sweden)

    Coulibaly Jean T

    2013-01-01

    Full Text Available Abstract Background Parasitological methods are widely used for the diagnosis of schistosomiasis. However, they are insensitive, particularly in areas of low endemicity, and labour-intensive. Immunoassays based on detection of anti-schistosome antibodies have the merit of high sensitivity and recently a rapid diagnostic test (RDT, incorporating Schistosoma mansoni cercarial transformation fluid (SmCTF for detection of anti-schistosome antibodies in blood has been developed. Here, we assessed the diagnostic performance of the SmCTF-RDT for S. mansoni and S. haematobium infections by comparing it with microscopy for egg detection. Methods A cross-sectional survey was carried out in Azaguié, south Côte d’Ivoire. 118 pre-school-aged children submitted two stool and two urine samples, which were subjected to the Kato-Katz and urine filtration methods for the detection of S. mansoni and S. haematobium eggs, respectively. Urine was also subjected to a commercially available cassette test for S. mansoni, which detects circulating cathodic antigen. A finger-prick blood sample was used for the SmCTF-RDT for detection of anti-S. mansoni and anti-S. haematobium antibodies. Results The prevalence of both anti-S. mansoni and anti-S. haematobium antibodies was more than three times higher than the prevalence of infection estimated by egg detection under a microscope. Using quadruplicate Kato-Katz as the reference standard for the diagnosis of S. mansoni infection, the sensitivity, negative predictive value (NPV, and positive predictive value (PPV of the SmCTF-RDT was 75.0%, 84.2% and 22.5%, respectively. When two urine filtrations were considered as the reference standard for the diagnosis of S. haematobium infection, the sensitivity, NPV and PPV of SmCTF-RDT was 66.7%, 94.9% and 5.1%, respectively. The specificity of SmCTF-RDT, when using egg-detection as the reference standard, was estimated to be 34.4%. This low specificity may be a reflection of the

  3. Total Spinal Block after Thoracic Paravertebral Block.

    Science.gov (United States)

    Beyaz, Serbülent Gökhan; Özocak, Hande; Ergönenç, Tolga; Erdem, Ali Fuat; Palabıyık, Onur

    2014-02-01

    Thoracic paravertebral block (TPVB) can be performed with or without general anaesthesia for various surgical procedures. TPVB is a popular anaesthetic technique due to its low side effect profile and high analgesic potency. We used 20 mL of 0.5% levobupivacaine for a single injection of unilateral TPVB at the T7 level with neurostimulator in a 63 year old patient with co-morbid disease who underwent cholecystectomy. Following the application patient lost consciousness, and was intubated. Haemodynamic instability was normalised with rapid volume replacement and vasopressors. Anaesthetic drugs were stopped at the end of the surgery and muscle relaxant was antagonised. Return of mucle strenght was shown with neuromuscular block monitoring. Approximately three hours after TPVB, spontaneous breathing started and consciousness returned. A total spinal block is a rare and life-threatening complication. A total spinal block is a complication of spinal anaesthesia, and it can also occur after peripheral blocks. Clinical presentation is characterised by hypotension, bradicardia, apnea, and cardiac arrest. An early diagnosis and appropriate treatment is life saving. In this case report, we want to present total spinal block after TPVB.

  4. Whole genome resequencing of the human parasite Schistosoma mansoni reveals population history and effects of selection

    Science.gov (United States)

    Crellen, Thomas; Allan, Fiona; David, Sophia; Durrant, Caroline; Huckvale, Thomas; Holroyd, Nancy; Emery, Aidan M.; Rollinson, David; Aanensen, David M.; Berriman, Matthew; Webster, Joanne P.; Cotton, James A.

    2016-01-01

    Schistosoma mansoni is a parasitic fluke that infects millions of people in the developing world. This study presents the first application of population genomics to S. mansoni based on high-coverage resequencing data from 10 global isolates and an isolate of the closely-related Schistosoma rodhaini, which infects rodents. Using population genetic tests, we document genes under directional and balancing selection in S. mansoni that may facilitate adaptation to the human host. Coalescence modeling reveals the speciation of S. mansoni and S. rodhaini as 107.5–147.6KYA, a period which overlaps with the earliest archaeological evidence for fishing in Africa. Our results indicate that S. mansoni originated in East Africa and experienced a decline in effective population size 20–90KYA, before dispersing across the continent during the Holocene. In addition, we find strong evidence that S. mansoni migrated to the New World with the 16–19th Century Atlantic Slave Trade. PMID:26879532

  5. Generalized Block Failure

    DEFF Research Database (Denmark)

    Jönsson, Jeppe

    2015-01-01

    Block tearing is considered in several codes as a pure block tension or a pure block shear failure mechanism. However in many situations the load acts eccentrically and involves the transfer of a substantial moment in combination with the shear force and perhaps a normal force. A literature study...... yield lines around the block leads to simple interaction formulas similar to other interaction formulas in the codes.......Block tearing is considered in several codes as a pure block tension or a pure block shear failure mechanism. However in many situations the load acts eccentrically and involves the transfer of a substantial moment in combination with the shear force and perhaps a normal force. A literature study...

  6. Recent advances in the diagnosis of Schistosoma infection: the detection of parasite DNA

    OpenAIRE

    Rabello Ana; Pontes Luís André; Dias-Neto Emmanuel

    2002-01-01

    As Schistosoma sp. control programs are chiefly based on treatment of infected population, adequate case finding has a crucial role. The available diagnostic methods are far from ideal, since the search for eggs in stools and the detection of circulating antigens lack sensitivity in low prevalence and post-treatment situations and antibody detection lacks specificity. In most endemic foci, repeated treatment of infected people leaves a number of non-diagnosed and consequently non-treated pers...

  7. Detection of Urinary Tract Pathology in Some Schistosoma haematobium Infected Nigerian Adults

    OpenAIRE

    Onile, O. S.; Awobode, H. O.; Oladele, V. S.; A. M. Agunloye; Anumudu, C. I.

    2016-01-01

    Screening for Schistosoma haematobium infection and its possible morbidity was carried out in 257 adult participants in Eggua community, Ogun State, Nigeria. Parasitological assessment for the presence of ova of S. haematobium in urine and abdominopelvic ultrasonographic examination for bladder and secondary kidney pathology were carried out. S. haematobium prevalence of 25.68% (66/257) was recorded among the participants. There was a significantly higher prevalence of 69.2% of urinary schist...

  8. Analysis of regulatory protease sequences identified through bioinformatic data mining of the Schistosoma mansoni genome

    Directory of Open Access Journals (Sweden)

    Minchella Dennis J

    2009-10-01

    Full Text Available Abstract Background New chemotherapeutic agents against Schistosoma mansoni, an etiological agent of human schistosomiasis, are a priority due to the emerging drug resistance and the inability of current drug treatments to prevent reinfection. Proteases have been under scrutiny as targets of immunological or chemotherapeutic anti-Schistosoma agents because of their vital role in many stages of the parasitic life cycle. Function has been established for only a handful of identified S. mansoni proteases, and the vast majority of these are the digestive proteases; very few of the conserved classes of regulatory proteases have been identified from Schistosoma species, despite their vital role in numerous cellular processes. To that end, we identified protease protein coding genes from the S. mansoni genome project and EST library. Results We identified 255 protease sequences from five catalytic classes using predicted proteins of the S. mansoni genome. The vast majority of these show significant similarity to proteins in KEGG and the Conserved Domain Database. Proteases include calpains, caspases, cytosolic and mitochondrial signal peptidases, proteases that interact with ubiquitin and ubiquitin-like molecules, and proteases that perform regulated intramembrane proteolysis. Comparative analysis of classes of important regulatory proteases find conserved active site domains, and where appropriate, signal peptides and transmembrane helices. Phylogenetic analysis provides support for inferring functional divergence among regulatory aspartic, cysteine, and serine proteases. Conclusion Numerous proteases are identified for the first time in S. mansoni. We characterized important regulatory proteases and focus analysis on these proteases to complement the growing knowledge base of digestive proteases. This work provides a foundation for expanding knowledge of proteases in Schistosoma species and examining their diverse function and potential as targets

  9. Developmental expression analysis and immunolocalization of a biogenic amine receptor in Schistosoma mansoni

    OpenAIRE

    El-Shehabi, Fouad; Vermeire, Jon J.; Yoshino, Timothy P.; Ribeiro, Paula

    2009-01-01

    A Schistosoma mansoni G-protein coupled receptor (SmGPCR) was previously cloned and shown to be activated by the biogenic amine, histamine. Here we report a first investigation of the receptor’s subunit organization, tissue distribution and expression levels in different stages of the parasite. A polyclonal antibody was produced in rabbits against the recombinant third intracellular loop (il3) of SmGPCR. Western blot studies of the native receptor and recombinant protein expressed in HEK293 c...

  10. Schistosoma mansoni proteins attenuate gastrointestinal motility disturbances during experimental colitis in mice

    Institute of Scientific and Technical Information of China (English)

    Nathalie; E; Ruyssers; Benedicte; Y; De; Winter; Joris; G; De; Man; Natacha; D; Ruyssers; Ann; J; Van; Gils; Alex; Loukas; Mark; S; Pearson; Joel; V; Weinstock; Paul; A; Pelckmans; Tom; G; Moreels

    2010-01-01

    AIM:To investigate the therapeutic effect of Schistosoma mansoni(S.mansoni) soluble worm proteins on gastrointestinal motility disturbances during experimental colitis in mice. METHODS:Colitis was induced by intrarectal injection of trinitrobenzene sulphate(TNBS) and 6 h later,mice were treated ip with S.mansoni proteins.Experiments were performed 5 d after TNBS injection.Inflammationwas quantified using validated inflammation parameters. Gastric emptying and geometric center were measured to assess in vivo...

  11. Recent advances in the diagnosis of Schistosoma infection: the detection of parasite DNA

    Directory of Open Access Journals (Sweden)

    Rabello Ana

    2002-01-01

    Full Text Available As Schistosoma sp. control programs are chiefly based on treatment of infected population, adequate case finding has a crucial role. The available diagnostic methods are far from ideal, since the search for eggs in stools and the detection of circulating antigens lack sensitivity in low prevalence and post-treatment situations and antibody detection lacks specificity. In most endemic foci, repeated treatment of infected people leaves a number of non-diagnosed and consequently non-treated persons, enough to maintain a persistent residue of 5 to 10% prevalence. In an attempt to surpass these diagnostic limitations we have developed a polymerase chain reaction (PCR for the detection of Schistosoma sp. in feces that, in a first population study, has shown to be more sensitive than three-repeated stool Kato-Katz examination. The PCR may constitute a valuable tool for the diagnosis of the Schistosoma sp. infection in special situations, when high sensitivity and specificity are required and infrastructure is available.

  12. Two sequential PCR amplifications for detection of Schistosoma mansoni in stool samples with low parasite load

    Directory of Open Access Journals (Sweden)

    Maria Cristina Carvalho do Espírito-Santo

    2012-10-01

    Full Text Available Schistosomiasis constitutes a major public health problem, with an estimated 200 million individuals infected worldwide and 700 million people living in risk areas. In Brazil there are areas of high, medium and low endemicity. Studies have shown that in endemic areas with a low prevalence of Schistosoma infection the sensitivity of parasitological methods is clearly reduced. Consequently diagnosis is often impeded due to the presence of false-negative results. The aim of this study is to present the PCR reamplification (Re-PCR protocol for the detection of Schistosoma mansoni in samples with low parasite load (with less than 100 eggs per gram (epg of feces. Three methods were used for the lysis of the envelopes of the S. mansoni eggs and two techniques of DNA extraction were carried out. Extracted DNA was quantified, and the results suggested that the extraction technique, which mixed glass beads with a guanidine isothiocyanate/phenol/chloroform (GT solution, produced good results. PCR reamplification was conducted and detection sensitivity was found to be five eggs per 500 mg of artificially marked feces. The results achieved using these methods suggest that they are potentially viable for the detection of Schistosoma infection with low parasite load.

  13. Genetic Variation between Biomphalaria alexandrina Snails Susceptible and Resistant to Schistosoma mansoni Infection

    Directory of Open Access Journals (Sweden)

    Suzanne M. F. El-Nassery

    2013-01-01

    Full Text Available Much effort has been made to control schistosomiasis infection in Egypt. However, enduring effects from such strategies have not yet been achieved. In this study, we sought to determine the genetic variability related to the interaction between Biomphalaria alexandrina snails and Schistosoma mansoni. Using RAPD-PCR with eight (10 mers random primers, we were able to determine the polymorphic markers that differed between snails susceptible and resistant to Schistosoma mansoni infection using five primers out of the eight. Our results suggest that the RAPD-PCR technique is an efficient means by which to compare genomes and to detect genetic variations between schistosomiasis intermediate hosts. The RAPD technique with the above-noted primers can identify genomic markers that are specifically related to the Biomphalaria alexandrina/Schistosoma mansoni relationship in the absence of specific nucleotide sequence information. This approach could be used in epidemiologic surveys to investigate genetic diversity among Biomphalaria alexandrina snails. The ability to determine resistant markers in Biomphalaria alexandrina snails could potentially lead to further studies that use refractory snails as agents to control the spread of schistosomiasis.

  14. Genetic variation between Biomphalaria alexandrina snails susceptible and resistant to Schistosoma mansoni infection.

    Science.gov (United States)

    El-Nassery, Suzanne M F; Abou-El-Naga, Iman F; Allam, Sonia R; Shaat, Eman A; Mady, Rasha F M

    2013-01-01

    Much effort has been made to control schistosomiasis infection in Egypt. However, enduring effects from such strategies have not yet been achieved. In this study, we sought to determine the genetic variability related to the interaction between Biomphalaria alexandrina snails and Schistosoma mansoni. Using RAPD-PCR with eight (10 mers) random primers, we were able to determine the polymorphic markers that differed between snails susceptible and resistant to Schistosoma mansoni infection using five primers out of the eight. Our results suggest that the RAPD-PCR technique is an efficient means by which to compare genomes and to detect genetic variations between schistosomiasis intermediate hosts. The RAPD technique with the above-noted primers can identify genomic markers that are specifically related to the Biomphalaria alexandrina/Schistosoma mansoni relationship in the absence of specific nucleotide sequence information. This approach could be used in epidemiologic surveys to investigate genetic diversity among Biomphalaria alexandrina snails. The ability to determine resistant markers in Biomphalaria alexandrina snails could potentially lead to further studies that use refractory snails as agents to control the spread of schistosomiasis.

  15. BLOCK H-MATRICES AND SPECTRUM OF BLOCK MATRICES

    Institute of Scientific and Technical Information of China (English)

    黄廷祝; 黎稳

    2002-01-01

    The block H-matrices are studied by the concept of G-functions, several concepts of block matrices are introduced. Equivalent characters of block H-matrices are obtained. Spectrum localizations claracterized by Gfunctions for block matrices are got.

  16. Block TERM factorization of block matrices

    Institute of Scientific and Technical Information of China (English)

    SHE Yiyuan; HAO Pengwei

    2004-01-01

    Reversible integer mapping (or integer transform) is a useful way to realize Iossless coding, and this technique has been used for multi-component image compression in the new international image compression standard JPEG 2000. For any nonsingular linear transform of finite dimension, its integer transform can be implemented by factorizing the transform matrix into 3 triangular elementary reversible matrices (TERMs) or a series of single-row elementary reversible matrices (SERMs). To speed up and parallelize integer transforms, we study block TERM and SERM factorizations in this paper. First, to guarantee flexible scaling manners, the classical determinant (det) is generalized to a matrix function, DET, which is shown to have many important properties analogous to those of det. Then based on DET, a generic block TERM factorization,BLUS, is presented for any nonsingular block matrix. Our conclusions can cover the early optimal point factorizations and provide an efficient way to implement integer transforms for large matrices.

  17. 日本血吸虫成虫肠道内容物中金属蛋白酶的鉴定%Identification of Metalloproteinase in Intestinal Contents Of Adult Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    袁仕善; 易新元; 曾宪芳; 张顺科; 黄跃龙; Larry MeReynolds

    2002-01-01

    目的鉴定日本血吸虫成虫肠道内容物中的金属蛋白酶.方法以明胶作底物,利用明胶-SDS-PAGE分离日本血吸虫成虫肠道内容物,将电泳后的凝胶于不同pH缓冲液和酶抑制剂中进行孵育,对其中的金属蛋白酶进行分析和鉴定.结果日本血吸虫成虫肠道内容物中存在降解明胶的金属蛋白酶,其最适pH值为7~9.金属蛋白酶抑制剂EDTA抑制其活性.电泳分离获得有活性的酶蛋白,该酶是血吸虫感染血清识别的弱抗原.结论日本血吸虫成虫肠道内容物存在金属蛋白酶.

  18. Construction and analysis of Sj - Rho GTPase DNA vaccine and subunit vaccine against Schistosoma japonicum%日本血吸虫Rho GTPase DNA疫苗及亚单位疫苗的构建和分析

    Institute of Scientific and Technical Information of China (English)

    张冉; 易新元; 曾宪芳; 黄跃龙; 蔡春; 张顺科; Larry McReynolds

    2003-01-01

    目的构建日本血吸虫大陆株Sj-Rho GTPase-like真核及原核表达重组质粒,并进行鉴定分析.方法用PCR法将Si-Rho GTPase-like基因从已剪切阳性克隆中扩增出来,亚克隆至pcDNA3.1和pGEX-5X-3中,分别经PCR、双酶切、测序、SDS-PAGE和Western blot等方法鉴定.结果PCR和测序均证明Sj-Rho GTPase-like基因疫苗构建成功,重组蛋白经SDS-PAGE电泳可观察到与预期分子量相应的条带,转印后可被水牛感染血清识别.结论成功构建了Si-Rho GTPase-like基因的两种重组载体,为保护性免疫研究打下基础.

  19. 日本血吸虫Calpain序列分析及DNA疫苗体系的构建%Sequence analysis and DNA vaccine preparation of Calpain,Schistosoma japonicum vaccine candidate molecule

    Institute of Scientific and Technical Information of China (English)

    周爱琴; 张仁利; 石淑华; 龙彩虹; 高世同; 林敏; 吴少庭

    2003-01-01

    目的探讨日本血吸虫疫苗候选分子-钙离子激活的中性蛋白激酶(Calpain)在抗日本血吸虫感染的保护性作用及其保护性免疫机制.方法从日本血吸虫成虫中提取RNA,用RT-PCR扩增Calpain含多个B,T细胞表位的片段,引物中包含BamHI和EcoRI的酶切位点,PCR扩增产物经纯化后TA克隆,转化的阳性TA克隆经PCR筛选后,液体培养大肠杆菌并回收质粒DNA,质粒DNA经BamHⅠ和EcoRⅠ双酶切,目的基因亚克隆到真核表达质粒pVAC载体,构建pVAC-Calpain真核表达体系,转化的阳性亚克隆经PCR筛选,液体培养大肠杆菌并回收pVAC-Calpain质粒DNA,质粒DNA经BamHⅠ和EcoRⅠ双酶切和DNA序列分析鉴定被亚克隆的Calpain基因.结果 RT-PCR从日本血吸虫RNA中扩增了453bp的Calpain基因,经BamHⅠ和EcoRⅠ双酶切和DNA序列分析鉴定Calpain基因被克隆到真核表达质粒pVAC载体.结论日本血吸虫疫苗候选分子Calpain的DNA疫苗体系的建立将有助于解析这个疫苗候选分子抗日本血吸虫感染的保护性免疫作用及保护性免疫机制.

  20. 日本血吸虫Calpain疫苗候选分子抗感染的免疫机制%Immunoprotective Mechanism of Calpain as Vaccine Candidate against Schistosoma japonicum Infection

    Institute of Scientific and Technical Information of China (English)

    张仁利; 黄达娜; 高世同; 耿艺介; 吴少庭; 胡章立; 张顺祥; 程锦泉

    2006-01-01

    目的 探讨日本血吸虫疫苗候选分子-钙离子激活的中性蛋白激酶(Calpain)的保护性免疫力和免疫保护性机制.方法 用重组纯化的Calpain抗原免疫BALB/c小鼠,制备抗Calpain血清,抗Calpain血清与机械转化的日本血吸虫童虫和来自于同种鼠激活的嗜酸性粒细胞共同培养,观察细胞对血吸虫童虫的粘附及对童虫的杀伤效果;RT-PCR分析Calpain抗原免疫BALB/c小鼠一氧化氮激酶(iNos.eNos.nNos)mRNA的表达;ELISA分析免疫小鼠体外培养脾细胞产生的细胞因子IFN-γ和IL-4.结果 重组Calpain抗原免疫小鼠后,产生了一个极高的抗Calpain特异性抗体,此抗体介导了同种鼠的嗜酸性粒细胞对日本血吸虫童虫的粘附,37℃、5%CO2培养48 h后,33.8%的日本血吸虫童虫被杀死,与控制组14.5%的自然死亡率相比有一个显著性的增高(P<0.01);RT-PCR测定重组Calpain抗原免疫1周小鼠iNos mRNA的表达显著性的增强,培养24 h脾细胞上清中IFN-γ浓度高于对照组.结论 Calpain特异性抗体介导了对日本血吸虫童虫的细胞毒作用,Calpain抗原能够诱导IFN-γ的产生,提示Calpain诱导Th1的免疫应答反应来抗日本血吸虫的感染.

  1. Research progress on Sj14-3-3 vaccine of Schistosoma japonicum%日本血吸虫Sj14-3-3疫苗研究进展

    Institute of Scientific and Technical Information of China (English)

    谭建蓉; 李文桂

    2014-01-01

    日本血吸虫病是由日本血吸虫引起的一类严重危害人类健康的人兽共患寄生虫病,研制疫苗防治该病是目前的研究热点.Sj14-3-3蛋白是一种有效的疫苗分子,该文就Sj14-3-3蛋白疫苗和核酸疫苗的研究进展进行综述.%Schistosomiasis japonica is a serious health-threatening parasitic zoonosis to human beings,which is caused by Schistosomajaponicum.Developing vaccines for schistosomiasis is a hot spot in the present studies.Sj14-3-3 protein is an effective vaccine.This article reviewed the progress on Sj14-3-3 protein vaccines and DNA vaccines.

  2. EFFECT OF pH ON THE MUTAGENIC POTENCY OF ICR-170 IN THE REPRODUCTIVE GLANDS OF FEMALE SCHISTOSOMA JAPONICUM%PH对ICR-170诱变日本血吸虫雌虫生殖腺的影响

    Institute of Scientific and Technical Information of China (English)

    蒋守富; 潘彩娥; 陆钦淹

    2003-01-01

    目的探讨溶液pH对诱变剂ICR-170诱变日本血吸虫雌虫生殖腺的影响程度.方法用5种pH值(pH 7.2-8.0)的ICR-170溶液处理日本血吸虫尾蚴30 min或45 min后,经皮肤接种小鼠观察雌虫诱变率和成虫回收率.结果溶液pH值对ICR-170的诱变作用影响较大,诱变率随着溶液pH值的上升而下降.10μg/ml 45 min组和15μg/ml 30 min组在pH 7.2时的雌虫诱变率分别为pH 7.8时的13倍和6倍,当pH>7.4时,2种浓度的ICR-170引起的诱变率基本接近.溶液pH值对于成虫回收率的影响程度远低于对诱变率的影响程度.结论在诱变日本血吸虫雌虫生殖腺时,ICR-170具有较强的pH依赖型特性.

  3. Experiment on Gypenosides in decreasing fibrosis in liver of mice infected with Schistosoma japonicum%阿魏减轻小鼠血吸虫性肝纤维化试验

    Institute of Scientific and Technical Information of China (English)

    田洁; 赵红梅

    2009-01-01

    血吸虫病的病理基础是虫卵肉芽肿。无虫卵或虽有虫卵而不发生或少发生肉芽肿,则不发生血吸虫病或病情较轻。国内外学者发现,在肉芽肿形成中自由基O-2起着重要作用,抗氧化剂能缩小肉芽肿体积。阿魏(Resina ferulae)是一种天然的抗氧化中药,具有抗肿瘤、降血脂等多种药理作用,但未见其抗血吸虫性肝纤维化的研究报道。笔者等用阿魏治疗试验小鼠血吸虫病,观察其对血吸虫性肝纤维化形成的影响。

  4. Proteomic analysis of vomit and excretory/secretory products of Schistosoma japonicum%日本血吸虫成虫呕吐和排泄分泌物的蛋白质组学分析

    Institute of Scientific and Technical Information of China (English)

    余传信; 赵飞; 殷旭仁; 肖迪; 张建中; 华万全; 钱春艳; 宋丽君; 王玠

    2010-01-01

    目的 探讨日本血吸虫成虫呕吐物和排泄分泌物的蛋白质组成,寻找高丰度蛋白分子.方法 将日本血吸虫成虫浸泡于无菌水中15~30 min,收集成虫的呕吐和排泄分泌物.通过蛋白质二维电泳,考马斯亮蓝染色与飞行质谱分析,确定各蛋白点的基因种类.结果 日本血吸虫成虫的呕吐及排泄分泌物的二维电泳获得1 012个蛋白斑点,456个斑点的蛋白质种类被质谱分析成功鉴定.这些蛋白可归为139种,其中76种为虫源性蛋白,主要是由一些与血吸虫生命代谢、生长发育、免疫调控相关的蛋白组成,其中烯醇化酶、70 kDa热休克蛋白(HSP70)、果糖二磷酸醛缩酶、三磷酸甘油醛脱氢酶、磷酸甘油酸激酶、谷胱甘肽转移酶、抑免蛋白、硫氧还蛋白过氧化物酶、14-3-3蛋白等含量丰富,是高丰度的排泄分泌蛋白.结论 本研究揭示了日本血吸虫成虫呕吐和排泄分泌物的蛋白组成,为进一步开发用于控制血吸虫病或其他寄生虫病的疫苗、新的治疗药物及诊断方法 创造了条件.

  5. 血吸虫成虫排泄分泌抗原的组成及免疫反应性分析%Analysis of the proteomics and immunoreactivities of excretory-secretory antigens of Schistosoma japonicum adult worms

    Institute of Scientific and Technical Information of China (English)

    华万全; 余传信; 王玠; 殷旭仁; 钱春艳

    2009-01-01

    目的 分析日本血吸虫成虫排泄分泌抗原的组成及其与血吸虫感染兔疗前、疗后配对血清的免疫反应特征.方法采用7 cm(pH5~8)的IPG和12% SDS-PAGE对血吸虫成虫排泄分泌抗原的组成进行双向凝胶电泳分析,并用免疫印迹试验检测排泄分泌抗原各蛋白点与疗前、疗后血清的反应性;采用PDQuest8.0图像分析软件对二维图像斑点进行分析,寻找差异反应蛋白点. 结果 血吸虫成虫排泄分泌抗原双向凝胶电泳图谱主要由215个蛋白斑点组成,其中16个斑点大、染色深,可能为高丰度的循环抗原;免疫印迹显示可被感染6周兔血清识别,但不能被治疗12周兔血清识别的蛋白斑点有84个,与感染6周兔血清的反应强度比与治疗12周兔血清的反应强度高2倍的蛋白点有38个. 结论 血吸虫成虫排泄分泌抗原中存在高丰度的能被短程抗体识别的抗原分子.

  6. 日本血吸虫(中国大陆株)Sj16基因的原核表达%PROKARYOTIC EXPRESSION OF Sj16 GENE OF SCHISTOSOMA JAPONICUM CHINESE STRAIN

    Institute of Scientific and Technical Information of China (English)

    卞国武; 余新炳; 吴忠道; 徐劲; 周俊梅; 单志新; 马长玲; 邵筱

    2001-01-01

    目的进一步研究日本血吸虫(中国大陆株)Sj16的免疫调节功能,丰富有关血吸虫免疫逃避知识.方法根据曼氏血吸虫Sm16基因已知序列设计合成一对引物,用PCR技术从日本血吸虫(中国大陆株)成虫cDNA文库中扩增Sj16基因;将Sj16基因定向克隆入pGEX-4T-1,转化感受态BL21/DE3菌;用酶切、PCR扩增鉴定筛选得到的重组阳性克隆.IPTG诱导pGEX-4T-1-Sj16转化的BL21/DE3菌,用SDS-PAGE和Wester-blot分析表达产物.结果从日本血吸虫(中国大陆株)成虫cDNA文库中获取Sj16基因,重组质粒中含有Sj16基因,成功构建日本血吸虫(中国大陆株)Sj16基因原核表达重组质粒pGEX-4T-1-Sj16,pGEX-4T-1-Sj16转化的BL21/DE3菌经IPTG诱导可表达与GST融合的蛋白(约40 kDa),抗GST抗体能特异性识别该蛋白.结论成功原核表达了Sj16蛋白,为深入研究Sj16的免疫调节功能,丰富有关血吸虫免疫逃避知识打下了基础.

  7. 放射配体受体分析法对日本血吸虫体内 5-HT受体的测定%The Detection of 5-HT Receptor by RLBA in Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    王丽平; 沈丽琳; 陈伟华

    2003-01-01

    采用放射配体-受体分析法(RLBA)测定了日本血吸虫体内5-羟色胺(5-HT)受体及螺环哌啶酮(spiroperidol)对5-HT受体与3H-5-HT结合反应的影响.结果证实:日本血吸虫体内存在5-HT受体,与配体5-HT可进行饱和的、特异的结合,其Scatchard图呈下凹曲线,表明5-HT受体的复杂性和不均一性.其最大结合容量(Bmax)为210.4 fmol/mg protein,解离常数(Kd)为0.585 pmol/L, 螺环哌啶酮对3H-5-HT与受体的结合无影响,计算Bmax为 270.3 fmol/mg protein, Kd为0.327 pmol/L,与前值比较,差异不显著(P>0.05).

  8. 肝片形吸虫长菲策吸虫及日本血吸虫DNA的多态性检测%Detection of DNA polymorphism among Fasciola hepatica, Fischoederius elongates and Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    胥全彬; 沈永林; 刘耀兴

    2001-01-01

    本试验用随机扩增DNA多态性技术对肝片形吸虫、长菲策吸虫和日本血吸虫虫体总DNA的多态性进行检测,结果筛选出三条具有种鉴别意义的RAPD谱带.这些特征性条带可望标记成探针用于粪便学检查中虫卵的鉴定.

  9. 左旋咪唑防御日本血吸虫尾蚴感染的实验研究%EXPERIMENTAL STUDIES ON EFFICACY OF LEVAMISOLE IN PREVENTION FROM SCHISTOSOMA JAPONICUM CERCARIA INFECTION

    Institute of Scientific and Technical Information of China (English)

    杨明瑾; 漏磊君; 严晓岚; 吴玲娟; 李思温

    2004-01-01

    目的探讨左旋咪唑预防日本血吸虫尾蚴感染的作用.方法盐酸左旋咪唑和左旋咪唑碱口服剂量为26.25 mg/kg,分别于小鼠感染前2天口服,连服7 d;涂肤剂为1.0%、2.0%、3.0%、5.0%和7.0%,分别于感染前2、1天和当天涂肤.停药后4周解剖,检获虫体.结果盐酸左旋咪唑水溶液和左旋咪唑碱水溶液分别口服,两者的减虫率都为0;5.0%的盐酸左旋咪唑于感染当天涂肤、7.0%盐酸左旋咪唑于感染前1天涂肤,减虫率均为100.0%;2.0%、3.0%和5.0%的左旋咪唑碱于感染前1天涂药,减虫率即可达到100.0%.结论左旋咪唑涂剂能防御日本血吸虫尾蚴的感染,左旋咪唑碱效果优于盐酸左旋咪唑.口服无效.

  10. TOXICITY OF HEAVY METALS CADMIUM AND ZINC TO CERCARIA OF SCHISTOSOMA JAPONICUM%重金属镉、锌对日本血吸虫尾蚴毒性作用的观察

    Institute of Scientific and Technical Information of China (English)

    卢致民; 唐宏炜; 张进顺; 纪伟华

    2005-01-01

    目的观察重金属镉、锌单独和混合存在时对日本血吸虫尾蚴的毒性作用. 方法分别将40~50个尾蚴置于不同浓度(100、10、1、0.1 mg/L)的镉、锌、镉/锌混合溶液中,20~25℃温箱培养,每隔30 min解剖显微镜下观察尾蚴形态变化与存活数量. 结果尾蚴暴露于浓度为10、100 mg/L的镉、锌、镉/锌混合物溶液后,在相同观察时间内,尾蚴断尾率、死亡率显著高于对照组(P<0.01或P<0.05);尾蚴断尾率、死亡率随镉、锌、镉/锌混合溶液浓度升高而升高,以100 mg/L的镉/锌混合溶液毒性作用最强. 结论重金属镉、锌无论是单独存在还是混合存在,对日本血吸虫尾蚴都有一定的毒性作用,并且浓度越高,毒性作用越强.

  11. In Vitro Effect of Photoactivated Hypericin on Anti-Schistosoma japonicum Adult Male Worms%光诱导金丝桃素体外抗日本血吸虫雄虫作用的观察

    Institute of Scientific and Technical Information of China (English)

    蔡茹; 佘新平; 王瑜; 龚唯; 张惠琴; 夏超明

    2014-01-01

    目的 观察光诱导后的金丝桃素对日本血吸虫雄虫的体外杀伤作用. 方法 80只昆明小鼠经腹部皮肤感染由单只阳性钉螺逸出的日本血吸虫尾蚴60~80条,6周后剖检,收集雄虫置入含有3 ml DMEM培养液的平皿内(10条/皿).每批次实验均分为:金丝桃素提取物或金丝桃素纯品(以下简称金丝桃素)不同浓度(0.1、0.2、0.5、1.0、1.5、2.0和2.5 μmol/L)光照组以及不同浓度非光照组,二甲基亚砜(DMSO)对照组和空白对照组.各实验均重复2~5次.将金丝桃素提取物或金丝桃素纯品及DMSO液分别加入含有血吸虫的培养皿中,恒温避光共孵育6h,再分别照射30、60、90和120 min后,37℃避光培养过夜(16 h).次晨,洗涤、更换无药物新鲜培养液,继续避光培养48 h,观察光诱导后的金丝桃素对血吸虫的杀伤作用.选取1.0、2.5和5.0 μmol/L金丝桃素光照60 min组虫体在解剖镜下观察并摄像;2.0 μmol/L金丝桃素光照60 min组虫体进行扫描电镜观察. 结果 各浓度金丝桃素纯品光照组的虫体均有不同程度的死亡,0.1 μmol/L金丝桃素纯品光照30 min组,虫体死亡率为20%;2μmol/L金丝桃素纯品光照90 min组和2.5 μmol/L金丝桃素纯品光照60 min组,虫体均100%死亡.相应的金丝桃素纯品各浓度无光照组、DMSO对照组和空白对照组,虫体均100%存活.解剖镜下观察,1.0、2.5和5.0 μmol/L金丝桃素纯品光照60 min组虫体均出现显著的痉挛性麻痹现象,表现为不同程度的缩短卷曲、体态僵硬和活动终止等改变;扫描电镜观察,2.0 μmol/L金丝桃素纯品光照60 min组虫体皮层出现肿胀、空泡形成、破裂、糜烂、剥落和感觉乳突丢失等损害,甚至皮层正常结构完全消失.金丝桃素纯品各浓度光照组的虫体死亡率和形态结构的损伤改变与金丝桃素浓度和光照时间有关. 结论 金丝桃素经光诱导后,具有显著的体外抗日本血吸虫雄虫作用.

  12. 日本血吸虫精氨酸酶基因启动子序列的扩增及序列分析%Amplification and Sequence Analysis of Schistosoma japonicum Arginase Novel Gene's Promoter Sequence

    Institute of Scientific and Technical Information of China (English)

    李孜; 余新炳; 吴忠道

    2005-01-01

    目的获取日本血吸虫(Sj)精氨酸酶(ARG)新基因的DNA编码序列及其启动子序列,实验验证ARG基因编码序列的完整性.方法 以日本血吸虫成虫 DNA做模板,PCR扩增 ARG基因编码区的DNA序列并测序;根据Sj ARG基因已扩增的 DNA序列设计2条巢式引物,用TaKaRa LA Taq PCR Cloning in vitro Kit,扩增 Sj ARG基因的启动子序列;对已扩增得到的序列进行 TATA盒的寻找以分析启动子序列的位置,实验验证我们曾获取的 Sj ARG新基因编码序列的完整性. 结果 扩增得到长约 1 000bp ARG基因 DNA序列 , Sj ARG基因 DNA序列内没有内含子,与 cDNA序列完全一致.对启动子序列扩增后,得到一略大于 250bp的序列,测序后分析发现其中有 36bp与 ARG基因 DNA序列的 5′端重叠.因此 ,将精氨酸酶基因的 DNA序列又向前延伸了 221bp,与前面得到的序列拼接后得到一条 1 486bp的总序列,将该序列在 NCBI上进行 ORF的寻找发现其最长的 ORF达 1 164bp,起始密码子在第 156位,终止密码子在第 1 319位 ,该起始密码子前 32位为 TATA盒的位置.因此,确定该 Sj ARG基因全长 cDNA编码序列长应为 1 164bp.结论 成功扩增获得了日本血吸虫精氨酸酶基因编码区的 DNA序列,其不含有内含子;并扩增得到了该基因的启动子序列,从而获得了 Sj ARG基因真正的全长 cDNA序列,为进一步的功能鉴定奠定了基础.

  13. A rapid and simple method of detection of Blepharisma japonicum using PCR and immobilisation on FTA paper

    Directory of Open Access Journals (Sweden)

    Hughes Jacqueline M

    2003-09-01

    Full Text Available Abstract Background The rapid expansion in the availability of genome and DNA sequence information has opened up new possibilities for the development of methods for detecting free-living protozoa in environmental samples. The protozoan Blepharisma japonicum was used to investigate a rapid and simple detection system based on polymerase chain reaction amplification (PCR from organisms immobilised on FTA paper. Results Using primers designed from the α-tubulin genes of Blepharisma, specific and sensitive detection to the equivalent of a single Blepharisma cell could be achieved. Similar detection levels were found using water samples, containing Blepharisma, which were dried onto Whatman FTA paper. Conclusion This system has potential as a sensitive convenient detection system for Blepharisma and could be applied to other protozoan organisms.

  14. (±)-Japonones A and B, two pairs of new enantiomers with anti-KSHV activities from Hypericum japonicum

    Science.gov (United States)

    Hu, Linzhen; Zhu, Hucheng; Li, Lei; Huang, Jinfeng; Sun, Weiguang; Liu, Junjun; Li, Hua; Luo, Zengwei; Wang, Jianping; Xue, Yongbo; Zhang, Yu; Zhang, Yonghui

    2016-06-01

    Two pairs of new enantiomers with unusual 5,5-spiroketal cores, termed (±)-japonones A and B [(±)-1 and (±)-2], were obtained from Hypericum japonicum Thunb. The absolute configurations of (±)-1 and (±)-2 were characterized by extensive analyses of spectroscopic data and calculated electronic circular dichroism (ECD) spectra, the application of modified Mosher’s methods, and the assistance of quantum chemical predictions (QCP) of 13C NMR chemical shifts. Among these metabolites, (+)-1 exhibited some inhibitory activity on Kaposi’s sarcoma associated herpesvirus (KSHV). Virtual screening of (±)-1 and (±)-2 were conducted using the Surflex-Dock module in the Sybyl software, and (+)-1 exhibited ability to bind with ERK to form key interactions with residues Lys52, Pro56, Ile101, Asp165, Gly167 and Val99.

  15. Fluorescence studies with malate dehydrogenase from rhizobium japonicum 3I1B-143 bacteroids: a two-tryptophan containing protein

    Science.gov (United States)

    Ghiron, Camillo A.; Eftink, Maurice R.; Waters, James K.; Emerich, David W.

    1990-05-01

    A number of fluorescence studies, both of trp residues and bound NADH, have been reported for porcine MDH. The large number of trp residues (6) complicates the interpretation of some studies. To circumvent this we have performed studies with a two tryptophan (per subunit) MDH from Rhizobium japonicum 311B-143 bacteroids. We have performed phase/modulation fluorescence lifetime measurements, as a function of temperature and added quencher KI, in order to resolved the 1.3 ns (blue) and 6.6 ns (red) contributions from the two classes of trp residues. Anisotropy decay studies have also been performed. The binding of NADH dynamically quenches the fluorescence of both tip residues, but, unlike mammalian cytoplasmic and mitochondrial MDH, there is not a large enhancement in fluorescence of bound NADH upon forming a ternary complex with either tartronic acid or D-malonate.

  16. The dual actions of angiogenesis and anti-apoptosis induced by an isolated fraction from Geum japonicum repair muscle ischemia.

    Science.gov (United States)

    Cheung, Winghong L; Cheng, Lei; Liu, Hongwei; Gu, Xuemei; Li, Ming

    2007-03-01

    The fundamental improvement of muscle ischemia requires the re-establishment of sufficient vessel network. Despite many kinds of drugs have been used for ischemia, effective angiogenic drug is very limited. Here, we reported the identification and isolation of a potent angiogenic fraction (angio-T) from Geum japonicum and assessment of its therapeutic effects on muscle ischemia by reconstituting the insufficient blood supply network and enhancing cell survival potential. It was demonstrated that angio-T not only significantly enhanced the proliferation of cultured HCAECs in vitro, but also significantly enhanced the survival potential of the myofibers at risk and neovascularization in ischemic muscles leading to reconstitution of these vessel networks, significant reduction of ischemic areas, and significant myofiber regeneration in ischemic area one week post-ischemia.

  17. Impact of elevated CO2 on carbohydrate and ureide concentrations in soybean inoculated with different strains of Bradyrhizobium japonicum

    Energy Technology Data Exchange (ETDEWEB)

    Bertrand, Annick; Prevost, Danielle; Juge, Christine; Chalifour, Francois-P.

    2011-07-15

    Over the past years, the CO2 concentration in the atmosphere has constantly increased and it is expected to continue; this climate change will lead to important impacts on the crops' productivity. The authors undertook a study to determine soybean's response to higher CO2 concentration and different strains of Bradyrhizobium japonicum. They observed that, under elevated CO2, additional photosynthates are produced and used for nodular growth and that ureides increased significantly in leaves but not much in nodules. The different strains resulted in different nodular yield and ureides concentration resulting in diverse responses to elevated CO2 concentration: the commercial strain showed a feedback inhibition of nodule activity while the indigenous strain 12NS14 with a better nodule activity had a positive feedback reaction with soybean growth. This study demonstrated that, under higher CO2 concentration, a better soybean production could be achieve with the strain 12NS14.

  18. Use of monoclonal antibodies prepared against Schistosoma mansoni hatching fluid antigens for demonstration of Schistosoma haematobium circulating egg antigens in urine.

    Science.gov (United States)

    Nibbeling, H A; Kahama, A I; Van Zeyl, R J; Deelder, A M

    1998-05-01

    A panel of 17 monoclonal antibodies (MAbs) against Schistosoma soluble egg antigens (SEAs) was produced from BALB/c mice immunized with antigens secreted/excreted by Schistosoma mansoni eggs. In this study, we demonstrate that 16 MAbs were reactive with S. haematobium SEA in addition to S. mansoni SEA. The MAbs were tested as potential immunodiagnostic reagents in a homologous sandwich ELISA format to detect circulating soluble egg antigens (CSEAs) in serum and urine samples of S. mansoni- or S. haematobium-infected individuals. When samples of S. mansoni-infected individuals were tested, none of these MAbs performed as good as the previously described S. mansoni-specific 114-5B1-A and 114-4D12-A MAbs. However, 11 MAbs (of the IgM isotype) detected CSEA in urine samples of S. haematobium-infected individuals. Three MAbs, 290-2E6-A, 291-3D5-A, and 291-5D5-A, were selected for a pilot study with 47 urine samples of S. haematobium-infected individuals from Kenya. The CSEA levels detected with each of these ELISAs showed a significant correlation with urinary egg counts (Spearman rho > 0.37, P 0.74, P < 0.001). Based on the 92% specificity and 90% sensitivity of the assay, the ELISA using MAb 290-2E6-A was found to be the most promising assay for immunodiagnosis of S. haematobium infections.

  19. Lesson Thirteen Trifascicular Block

    Institute of Scientific and Technical Information of China (English)

    鲁端; 王劲

    2005-01-01

    @@ A complete trifascicular block would result in complete AV block. The idio ventricular rhythm has a slower rate and a wide QRS complex because the pacemaker is located at the peripheral part of the conduction system distal to the sites of the block1. Such a rhythm may be difficult to differentiate from bifascicular or bundle branch block combined with complete block at a higher level such as the AV node or His bundle2. Besides a slower ventricular rate, a change in the morphology of the QRS complex from a previous known bifascicular pattern would be strongly suggestive of a trifascicular origin of the complete AV block3. A His bundle recording is required for a definitive diagnosis, however.

  20. Isovaleryl-homoserine lactone, an unusual branched-chain quorum-sensing signal from the soybean symbiont Bradyrhizobium japonicum.

    Science.gov (United States)

    Lindemann, Andrea; Pessi, Gabriella; Schaefer, Amy L; Mattmann, Margrith E; Christensen, Quin H; Kessler, Aline; Hennecke, Hauke; Blackwell, Helen E; Greenberg, E Peter; Harwood, Caroline S

    2011-10-01

    Many species of Proteobacteria communicate by using LuxI-LuxR-type quorum-sensing systems that produce and detect acyl-homoserine lactone (acyl-HSL) signals. Most of the known signals are straight-chain fatty acyl-HSLs, and evidence indicates that LuxI homologs prefer fatty acid-acyl carrier protein (ACP) over fatty acyl-CoA as the acyl substrate for signal synthesis. Two related LuxI homologs, RpaI and BtaI from Rhodopseudomonas palustris and photosynthetic stem-nodulating bradyrhizobia, direct production of the aryl-HSLs p-coumaroyl-HSL and cinnamoyl-HSL, respectively. Here we report that BjaI from the soybean symbiont Bradyrhizobium japonicum USDA110 is closely related to RpaI and BtaI and catalyzes the synthesis of isovaleryl-HSL (IV-HSL), a branched-chain fatty acyl-HSL. We show that IV-HSL induces expression of bjaI, and in this way IV-HSL functions like many other acyl-HSL quorum-sensing signals. Purified histidine-tagged BjaI was an IV-HSL synthase, which was active with isovaleryl-CoA but not detectably so with isovaleryl-ACP. This suggests that the RpaI-BtaI-BjaI subfamily of acyl-HSL synthases may use CoA- rather than ACP-linked substrates for acyl-HSL synthesis. The bjaI-linked bjaR(1) gene is involved in the response to IV-HSL, and BjaR(1) is sensitive to IV-HSL at concentrations as low as 10 pM. Low but sufficient levels of IV-HSL (about 5 nM) accumulate in B. japonicum culture fluid. The low levels of IV-HSL synthesis have likely contributed to the fact that the quorum-sensing signal from this bacterium has not been described elsewhere.

  1. A multivariate analysis of the relationship between work ability and S. japonicum infection in Dongting Lake Region, in China Análise multivariada da relação entre capacidade de trabalho e infecção por S. japonicum na região dos lagos de Dongting, China

    Directory of Open Access Journals (Sweden)

    Li Yuesheng

    1993-08-01

    Full Text Available A cross-sectional case-control study on the association between the reduced work ability and S. japonicum infection was carried out in a moderate endemic area for schistosomiasis japonica in the southern part of Dongting lake in China. A total of 120 cases with reduced work ability and 240 controls paired to the case by age, sex, occupation and without reduced work ability, participated in the study. The mean age for individuals was 37.6 years old (21-60, the ratio of male: female was 60:40, the prevalence of S. japonicum in the individuals was 28.3%. The results obtained in this study showed that the infection of S. japonicum in case and control groups was 49.2% (59/120 and 17.9% (43/240, respectively. Odds ratio for reduced work ability among those who had schistosomiasis was 4.34 (95%, confidence interval was 2.58-7.34, and among those who had S. japonicum infection (egg per gram > 100 was up to 12.67 (95%, confidence interval was 3.64-46.39. After odds ratio was adjusted by multiple logistic regression, it was confirmed that heavier intensity of S. japonicum infection and splenomegaly due to S. japonicum infection were the main risk factors for reduced work ability in the population studied.Um estudo seccional de casos controles da associação entre a capacidade reduzida para o trabalho e a infecção por S. japonicum foi levada a efeito em região moderadamente endêmica para esquistossomose japônica na parte sul do lago Dongting, China. Um total de 120 casos com redução da capacidade de trabalho e 240 controles pareados no que diz respeito a idade, sexo, ocupação sem redução da capacidade de trabalho. A idade média dos pacientes foi 37,6 anos (21-60 e a relação masculino:feminino foi 60:40. A prevalência do S. japonicum foi de 28,3%. Os resultados obtidos neste estudo mostraram que a infecção nos casos e no grupo controle foi 49,2% (59/120 e 17,9% (43/240 respectivamente. A média para redução da capacidade de trabalho

  2. An L-glucitol oxidizing dehydrogenase from Bradyrhizobium japonicum USDA 110 for production of D-sorbose with enzymatic or electrochemical cofactor regeneration.

    Science.gov (United States)

    Gauer, Sabrina; Wang, Zhijie; Otten, Harm; Etienne, Mathieu; Bjerrum, Morten Jannik; Lo Leggio, Leila; Walcarius, Alain; Giffhorn, Friedrich; Kohring, Gert-Wieland

    2014-04-01

    A gene in Bradyrhizobium japonicum USDA 110, annotated as a ribitol dehydrogenase (RDH), had 87 % sequence identity (97 % positives) to the N-terminal 31 amino acids of an L-glucitol dehydrogenase from Stenotrophomonas maltophilia DSMZ 14322. The 729-bp long RDH gene coded for a protein consisting of 242 amino acids with a molecular mass of 26.1 kDa. The heterologously expressed protein not only exhibited the main enantio selective activity with D-glucitol oxidation to D-fructose but also converted L-glucitol to D-sorbose with enzymatic cofactor regeneration and a yield of 90 %. The temperature stability and the apparent K m value for L-glucitol oxidation let the enzyme appear as a promising subject for further improvement by enzyme evolution. We propose to rename the enzyme from the annotated RDH gene (locus tag bll6662) from B. japonicum USDA as a D-sorbitol dehydrogenase (EC 1.1.1.14).

  3. Blocked Urethral Valves

    Science.gov (United States)

    ... Blocked Urethral Valves Health Issues Listen Español Text Size Email Print Share Blocked Urethral Valves Page Content Article Body Urine leaves the bladder through a tube called the urethra, which in boys passes through the penis. Rarely, small membranes form across the urethra in ...

  4. Types of Heart Block

    Science.gov (United States)

    ... P wave as it normally would. If an electrical signal is blocked before it reaches the ventricles, they won't contract and pump blood to the lungs and the rest of the body. Second-degree heart block is divided into two ...

  5. Related Drupal Nodes Block

    NARCIS (Netherlands)

    Van der Vegt, Wim

    2010-01-01

    Related Drupal Nodes Block This module exposes a block that uses Latent Semantic Analysis (Lsa) internally to suggest three nodes that are relevant to the node a user is viewing. This module performs three tasks. 1) It periodically indexes a Drupal site and generates a Lsa Term Document Matrix. Inde

  6. Dicty_cDB: CHS126 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available SJS Schistosoma japonicum cDNA, mRNA sequence. 56 8e-04 1 CV682907 |CV682907.1 service...43_H06_sjs2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 8e-04 1 CV682854 |CV682854.1 service

  7. Dicty_cDB: SHD168 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service...43_H06_sjs2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service

  8. The Block Neighborhood

    CERN Document Server

    Arrighi, Pablo

    2010-01-01

    We define the block neighborhood of a reversible CA, which is related both to its decomposition into a product of block permutations and to quantum computing. We give a purely combinatorial characterization of the block neighborhood, which helps in two ways. First, it makes the computation of the block neighborhood of a given CA relatively easy. Second, it allows us to derive upper bounds on the block neighborhood: for a single CA as function of the classical and inverse neighborhoods, and for the composition of several CAs. One consequence of that is a characterization of a class of "elementary" CAs that cannot be written as the composition of two simpler parts whose neighborhoods and inverse neighborhoods would be reduced by one half.

  9. Ultrastructural analysis of miltefosine-induced surface membrane damage in adult Schistosoma mansoni BH strain worms.

    Science.gov (United States)

    Bertão, Humberto Gonçalves; da Silva, Renata Alexandre Ramos; Padilha, Rafael José R; de Azevedo Albuquerque, Mônica Camelo Pessôa; Rádis-Baptista, Gandhi

    2012-06-01

    Schistosomiasis is an infectious parasitic disease caused by helminths from the genus Schistosoma; it affects over 200 million people globally and is endemic in 70 countries. In Brazil, 6 million individuals are infected with Schistosoma mansoni. Furthermore, as the prevalence of S. mansoni infections is increasing, approximately 26 million citizens in 19 Brazilian states are at risk for infection. Schistosomiasis disease control involves predominately the administration of a single drug, praziquantel. Although praziquantel exhibits chemotherapeutic efficacy and safety, its massive use in endemic zones, the possibility of the emergence of drug-resistant Schistosoma parasites, and the lack of another efficacious antischistosomal drug demand the discovery of new schistosomicidal compounds. First developed as anti-tumor drug, miltefosine is an alkylphospholipid derivative that exhibits bioactivity against Leishmania and Trypanosoma parasites, free-living protozoa, bacteria, and fungi. With its anti-parasite activity, miltefosine was the first orally administered drug against visceral and cutaneous leishmaniasis approved. Previously, by means of the MTT cytotoxic assay and a DNA fragmentation test, we verified that, at doses of 100 and 200 μM (40 and 80 μg/mL), miltefosine exhibited in vitro schistosomicidal activity against adult S. mansoni worms. Here, we present ultrastructural evidence of rapid, severe miltefosine-induced surface membrane damage in S. mansoni following drug treatment. The number of dead parasites was concentration- and time-dependent following miltefosine treatment. At a miltefosine concentration of 200 μM (∼80 μg/mL), in vitro parasite killing was initiated as early as 3 h post-incubation, and it was maximal after 24 h of treatment. The parasite death was preceded by progressive surface membrane damage, characterized by tegument peeling, spine reduction and erosion, blister formation and rupture, and the emergence of holes. According to our

  10. Schistosoma mansoni Sambon, 1907: comparative morphologica studies of some Brazilian strains Schistosoma mansoni Sambon, 1907: estudos comparativos da morfologia de algumas cepas brasileiras

    Directory of Open Access Journals (Sweden)

    José Roberto Machado-Silva

    1995-10-01

    Full Text Available The morphology of Schistosoma mansoni adult male worms from three strains which have been maintained in albino mice for several generations, was compared to a strain that has been isolated from the natural host Nectomys squamipes (Rodentia: Muridae captured in Sumidouro (Rio de Janeiro State and have been maintained in the same sylvatic rodent under laboratory conditions. Total length of specimens, distance between suckers, the number of testes and extention of testes grouping were the taxonomic characters analysed. The worms recovered from N. squamipes showed expressive differences (pA morfologia de vermes adultos machos de tres cepas de Schistosoma mansoni, mantidas por várias gerações em camundongos albinos, foi comparada com uma cepa isolada do hospedeiro natural Nectomys squamipes (Rodentia: Muridae e mantida, em laboratório, neste mesmo roedor silvestre. Como caracteres taxonômicos foram analisados o comprimento total, o numero de testículos, a distância entre as ventosas e a distância ocupada pelos testículos nos espécimes. Os vermes recuperados de N. squamipes apresentaram diferenças significativas (p<0,01 em relação às outras cepas para quaisquer caracteres morfológicos estudados. As cepas mantidas em camundongos apresentaram diferença estatística em vários caracteres (p<0,01. Alguns vermes adultos além da disposição normal dos testículos, apresentavam também uma localização atípica destas glândulas sexuais. Conclui-se que a morfologia dos vermes adultos pode ser utilizada para caracterizar cepas de S. mansoni e que as passagens sucessivas de uma cepa em camundongos não induzem a alterações morfológicas nos vermes adultos.

  11. Block copolymer battery separator

    Science.gov (United States)

    Wong, David; Balsara, Nitash Pervez

    2016-04-26

    The invention herein described is the use of a block copolymer/homopolymer blend for creating nanoporous materials for transport applications. Specifically, this is demonstrated by using the block copolymer poly(styrene-block-ethylene-block-styrene) (SES) and blending it with homopolymer polystyrene (PS). After blending the polymers, a film is cast, and the film is submerged in tetrahydrofuran, which removes the PS. This creates a nanoporous polymer film, whereby the holes are lined with PS. Control of morphology of the system is achieved by manipulating the amount of PS added and the relative size of the PS added. The porous nature of these films was demonstrated by measuring the ionic conductivity in a traditional battery electrolyte, 1M LiPF.sub.6 in EC/DEC (1:1 v/v) using AC impedance spectroscopy and comparing these results to commercially available battery separators.

  12. Hawaii Census 2000 Blocks

    Data.gov (United States)

    U.S. Environmental Protection Agency — This data layer represents Census 2000 demographic data derived from the PL94-171 redistricting files and SF3. Census geographic entities include blocks, blockgroups...

  13. Steroidal neuromuscular blocking agents

    NARCIS (Netherlands)

    Wierda, JMKH; Mori, K; Ohmura, A; Toyooka, H; Hatano, Y; Shingu, K; Fukuda, K

    1998-01-01

    Since 1964 approximately 20 steroidal neuromuscular blocking agents have been evaluated clinically. Pancuronium, a bisquaternary compound designed on the drawingboard, was the first steroidal relaxant introduced into clinical practice worldwide in the 1970's. Although a major improvement, pancuroniu

  14. Superalloy Lattice Block Structures

    Science.gov (United States)

    Nathal, M. V.; Whittenberger, J. D.; Hebsur, M. G.; Kantzos, P. T.; Krause, D. L.

    2004-01-01

    Initial investigations of investment cast superalloy lattice block suggest that this technology will yield a low cost approach to utilize the high temperature strength and environmental resistance of superalloys in lightweight, damage tolerant structural configurations. Work to date has demonstrated that relatively large superalloy lattice block panels can be successfully investment cast from both IN-718 and Mar-M247. These castings exhibited mechanical properties consistent with the strength of the same superalloys measured from more conventional castings. The lattice block structure also accommodates significant deformation without failure, and is defect tolerant in fatigue. The potential of lattice block structures opens new opportunities for the use of superalloys in future generations of aircraft applications that demand strength and environmental resistance at elevated temperatures along with low weight.

  15. Structure-based study of natural products with anti-Schistosoma activity.

    Science.gov (United States)

    Akachukwu, Ibezim; Olubiyi, Olujide Oludayo; Kosisochukwu, Ata; John, Mbah Chika; Justina, Nwodo Ngozi

    2017-01-19

    Background Schistosomiasis is a parasitic protozoal disease caused by flatworms of the genus Schistosoma. Although the disease threatens millions of lives, it is still at the top list of neglected tropical diseases and praziquantel, the only common schistosocidal drug in use, has records of decreasing efficiency and cases of resistance. Also, reports revealed that people in the rural areas, who are most affected, rely mostly on traditional herbal medicines because of limited access to modern healthcare. The use of computers in drug development has become a routine practice because they are cost and time effective.

  16. Evolution of sarcoma 180 (ascitic tumor in mice infected with Schistosoma mansoni

    Directory of Open Access Journals (Sweden)

    Fausto Edmundo Lima Pereira

    1986-03-01

    Full Text Available Mice infected with 60 cercariae of Schistosoma mansoni were more resistant to the sarcoma 180 ascites tumor. Tumor inoculation was performed 50 days after schistosoma infection and the animals were observed and weighed at 48 hours intervals for development and progression of malignancy. In infected mice the weight gain (ascites formation started later and was shorter than in uninfected Controls. Also, the number of tumor cells into the peritoneal cavity 72h after tumor implantation was shorter in infected group than incontrols. This in creased resistance against a transplantable tumor probably is related to the effect of endotoxin on tumoricidal activity of macrophages activated by the infection. The immunodepression induced by Schistosoma mansoni infection enhances the proliferation of endogenous bacteria increasing the amount of endotoxin absorbed from the gut.Camundongos infectados com 60 cercárias de Schistosoma mansoni tomaram-se mais resistentes ao sarcoma 180 na forma de tumor ascítico. A inoculação das células tumorais foi feita no 50º dia de infecção e a evolução do tumor foi acompanhada através dapesagem dos animais com intervalos de 48 horas. Nos camundongos infectados o ganho de peso (formação da ascite começou mais tarde e foi menor do que nos controles não infectados. Também o número de células tumorais na cavidade peritoneal 72 horas após a implantação do tumor foi menor no grupo infectado. Este aumento de resistência a um tumor transplantávelpossivelmente está relacionado ao efeito de endotoxinas sobre a atividade tumoricida dos macrofagos ativados pela infecção. A imunossupressão induzida pela infecção favorece a proliferação de bactérias da flora endógena aumentando a quantidade de endotoxinas absorvidas pelo intestino.

  17. Sequential histological changes in Biomphalaria glabrata during the course of Schistosoma mansoni infection

    Directory of Open Access Journals (Sweden)

    Queli Teixeira Lemos

    2001-07-01

    Full Text Available Biomphalaria glabrata, highly susceptible to Schistosoma mansoni, were seen to shed less and less cercariae along the time of infection. Histological examination kept a close correlation with this changing pattern of cercarial shedding, turning an initial picture of no-reaction (tolerance gradually into one of hemocyte proliferation with formation of focal encapsulating lesions around disintegrating sporocysts and cercariae, a change that became disseminated toward the 142nd day post miracidial exposure. Findings were suggestive of a gradual installation of acquired immunity in snails infected with S. mansoni.

  18. Specific immunoglobulin measurements related to exposure and resistance to Schistosoma mansoni infection in Sudanese canal cleaners

    DEFF Research Database (Denmark)

    Satti, M.Z.; Lind, Peter; Vennervald, B.J.;

    1996-01-01

    The present work comprises a longitudinal study of Schistosoma mansoni infection in occupationally hyper-exposed canal cleaners in the Sudan and the influence of chemotherapy on humoral immune parameters. The study groups included chronically infected canal cleaners (n = 19), newly recruited canal...... was used to detect specific IgE and IgG subclasses in response to whole worm antigen (WWH) and soluble egg antigen (SEA) before and 3 months after praziquantel treatment in the groups of canal cleaners and before and 1 year after treatment in normally exposed adults. When intensity of infection...

  19. Cistitis por schistosoma haematobium, presentación de un caso

    OpenAIRE

    2014-01-01

    No es frecuente en nuestro medio la infección por Schistosoma. haematobium, pero existen todos los elementos de la triada ecológica para su transmisión. Se presenta el caso de un paciente masculino, de raza mestiza, con 46 años de edad y procedencia rural, que refiere dolores lumbares al orinar, disuria, tenesmo vesical y polaquiuria durante el último año. Al examen físico se encontró el abdomen discretamente doloroso a la palpación superficial y profunda en hipogastrio y fosas lumbares. Se t...

  20. Schistosoma mansoni: effects of anesthetics and antimonial drugs on worm shift in the mouse

    Directory of Open Access Journals (Sweden)

    José Renan da Cunha-Melo

    1986-08-01

    Full Text Available Mice experimentally infected with Schistosoma mansoni were injected with sodium thiopental or sodium antimonyl gluconate (Triostib R, or submitted to halothane inhalation, with or without a previous injection of thiopental. Data obtained showed that halothane and thiopental induce worm shift to the liver (99 and 76%, respectively. Sodium gluconate and antimonium (Triostib R shifted 52% of worms towards the liver. These results seem to indicate that the use of antimonium would be unnecessary, when surgical removal of schistosomules is carried out through the extracorporeal filtration technique, in patients with portal hypertension.

  1. The feasibility of MS and advanced data processing for monitoring Schistosoma mansoni infection

    DEFF Research Database (Denmark)

    Balog, Crina I.A.; Alexandrov, Theodore; Derks, Rico J.

    2010-01-01

    Sensitive diagnosis, monitoring of disease progression and the evaluation of chemotherapeutic interventions are of prime importance for the improvement of control and prevention strategies for Schistosomiasis. The aim of the present study was to identify novel markers of Schistosoma mansoni infec...

  2. Suppression of basophil histamine release and other IgE-dependent responses in childhood Schistosoma mansoni/hookworm coinfection

    DEFF Research Database (Denmark)

    Pinot de Moira, Angela; Fitzsimmons, Colin M; Jones, Frances M;

    2014-01-01

    in Ugandan schoolchildren coinfected with Schistosoma mansoni and hookworm. METHODS: Skin prick test (SPT) sensitivity to house dust mite allergen (HDM) and current wheeze were assessed pre-anthelmintic treatment. Nonspecific (anti-IgE), helminth-specific, and HDM-allergen-specific basophil histamine release...

  3. Phenotypic characterization of Schistosoma mansoni adult wormsrecovered from undernourished mice: a morphometric study focusing on the reproductive system

    Directory of Open Access Journals (Sweden)

    Neves Renata Heisler

    2002-01-01

    Full Text Available A morphometric study focusing on some features of the reproductive system of Schistosoma mansoni adult worms was performed, aiming to complete previously reported data concerning the effects of undernourishment of the host on the parasites. Male worms were significantly affected (p<0.05 regarding the testicular lobes.

  4. Humoral and cellular immune responses induced in mice by purified iridoid mixture that inhibits penetration of Schistosoma mansoni cercariae upon topical treatment of mice tails.

    Science.gov (United States)

    Bahgat, Mahmoud; Shalaby, Nagwa M M; Ruppel, Andreas; Maghraby, Amany S

    2005-08-01

    When tested for possible blocking effect on the cercarial, serine proteinase, elastase (CE) activity, an iridoid mixture extracted from leaves of Citharexylum quadrangular abolished 31% of the enzyme activity at final concentration 15 microg. When formulated in jojoba oil and applied to mice tails followed by infection with Schistosoma mansoni cercariae, the iridoid mixture blocked cercarial penetration and caused significant reducetion (94%; P < 0.05) in worm burden in treated mice in comparison to controls. Also, immunomodulatory effects of iridoid mixture, iridoid-treated S. mansoni worm homogenate on mice were studied by measuring IgG and IgM levels against E. coli lysates (ECL), solube S. mansoni worm antigenic preparation (SWAP) and cancer bladder homogenates (CBH) as antigens by ELISA. Cellular immune responses were studied by calculating mean percent of CD4+, CD8(+)-T, B-mesenteric lymph node cells (MLNC) and CD4+, CD8(+)-T thymocytes by direct immunofluorescence staining in treated mice as compared to untreated homogenate given mice or untreated mice. Injecting mice with serial dilutions of iridoid mixture resulted in fluctuation, peaks and troughs, in both IgG and IgM responses against the above mentioned antigens. 1st and 2nd immunizations with iridoid mixture treated homogenate resulted in significantly elevated (P < 0.05). IgM and IgG levels against the 3 used antigens in comparison with sera from control mice. Immunized mice with homogenate treated with iridoid mixture showed a significant increase (P < 0.05) in CD4+T thymocytes, a non significant increase in CD8+T thymocytes, a significant increase (P < 0.05) in CD4+T lymphocytes (MLNC) and a non significant increase in CD8+ T- and B-lymphocytes (MLNC) compared with mice immunized with untreated homogenate or non-injected normal mice.

  5. Papain-Based Vaccination Modulates Schistosoma mansoni Infection-Induced Cytokine Signals.

    Science.gov (United States)

    Abdel Aziz, N; Tallima, H; Hafez, E A; El Ridi, R

    2016-02-01

    We have previously shown that immunization of outbred rodents with cysteine peptidases-based vaccine elicited type 2-biased immune responses associated with consistent and reproducible protection against challenge Schistosoma mansoni. We herein start to elucidate the molecular basis of C57BL/6 mouse resistance to S. mansoni following treatment with the cysteine peptidase, papain. We evaluated the early cytokine signals delivered by epidermal, dermal, and draining lymph node cells of naïve, and S. mansoni -infected mice treated 1 day earlier with 0 or 50 μg papain, or immunized twice with papain only (10 μg/mouse), papain-free recombinant S. mansoni glyceraldehyde 3-phosphate dehydrogenase and 2-Cys peroxiredoxin peptide (10 and 15 μg/mouse, respectively = antigen Mix), or papain-adjuvanted antigen Mix. Schistosoma mansoni infection induced epidermal and lymph node cells to release type 1, type 2 and type 17 cytokines, known to counteract each other. Expectedly, humoral immune responses were negligible until patency. Papain pretreatment or papain-based vaccination diminished or shut off S. mansoni infection early induction of type 1, type 17 and type 2 cytokines except for thymic stromal lymphopoietin and programmed the immune system towards a polarized type 2 immune milieu, associated with highly significant (P < 0.005 - <0.0001) resistance to S. mansoni infection.

  6. Mefloquine interferes with glycolysis in schistosomula of Schistosoma mansoni via inhibition of enolase.

    Science.gov (United States)

    Manneck, Theresia; Keiser, Jennifer; Müller, Joachim

    2012-04-01

    The antimalarial drug mefloquine has promising antischistosomal properties killing haematophagous adult schistosomes as well as schistosomula. The mode of action and involved drug targets of mefloquine in Schistosoma mansoni schistosomula are unknown. In order to identify mefloquine-binding proteins and thus potential drug targets, mefloquine affinity chromatography with S. mansoni schistosomula crude extracts was performed. We found one specific mefloquine-binding protein that was identified by mass spectrometry as the glycolytic enzyme enolase (Q27877). Enolase activity assays were performed on schistosomula crude extracts and on the recombinant enolase Q27877 expressed in Escherichia coli. In schistosomula crude extracts enolase activity was inhibited by mefloquine and by the enolase inhibitor sodium fluoride, while activity of the recombinant enolase was not affected. In contrast to enolase from crude extracts, recombinant Q27877 did not bind to mefloquine-agarose. Using isothermal microcalorimetry, we next investigated the metabolic inhibition of mefloquine and 3 known glycolytic inhibitors in Schistosoma spp., namely sodium fluoride, 3-bromopyruvate and menadione on schistosomula in the presence or absence of glucose. We found that in the presence of glucose, schistosomula were less affected by mefloquine, sodium fluoride and 3-bromopyruvate, whereas glucose had no protective effect when schistosomula had been exposed to menadione. These results suggest a potential role of mefloquine as an inhibitor of glycolysis, at least in stages where other targets like haem degradation are not relevant.

  7. Cistitis por schistosoma haematobium, presentación de un caso

    Directory of Open Access Journals (Sweden)

    Ramón Zaragoza Durañona

    2014-08-01

    Full Text Available No es frecuente en nuestro medio la infección por Schistosoma. haematobium, pero existen todos los elementos de la triada ecológica para su transmisión. Se presenta el caso de un paciente masculino, de raza mestiza, con 46 años de edad y procedencia rural, que refiere dolores lumbares al orinar, disuria, tenesmo vesical y polaquiuria durante el último año. Al examen físico se encontró el abdomen discretamente doloroso a la palpación superficial y profunda en hipogastrio y fosas lumbares. Se trató como una infección urinaria sin mejoría. Los complementarios arrojaron discreta eosinofilia y reflujo vesicoureteral grado IV en la uretrocistografía retrógrada y miccional. La cistoscopia videoasistida reportó, a nivel del trígono, la mucosa vesical enrojecida con estructuras en forma de granos de arroz de aspecto puntiforme; la biopsia informó huevos de esquistosomas y cambios metaplásicos en el epitelio vesical. Se trató con praziquantel y desaparecieron los síntomas. A los seis meses los complementarios resultaron normales y el paciente está asintomático. Se concluye en la importancia de considerar la infección por Schistosoma haematobium entre los diagnósticos de la infección urinaria refractaria al tratamiento

  8. The schistosoma-specific antibody response after treatment in non-immune travellers.

    Science.gov (United States)

    Duus, Liv Marie; Christensen, Anders Vittrup; Navntoft, Dorte; Tarp, Britta; Nielsen, Henrik V; Petersen, Eskild

    2009-01-01

    Egg detection is the gold standard in diagnosing and controlling treatment in schistosomiasis, but sensitivity is poor in lightly infected individuals, whereas Schistosoma-specific antibodies are more sensitive. The purpose of the study was to evaluate use of Gut Associated Antigen (GAA) and Membrane Bound Antigen (MBA) assays in assessment of treatment efficacy and number of treated non-immune individuals with signs of treatment failure. In a retrospective study, residents in Denmark diagnosed with positive Schistosoma antibodies in the period 1987 - 2004 were offered follow-up including analyses for GAA, MBA, IgE and eosinophil count. Among 98 patients with positive antibody at time of diagnosis, 73 were examined for eggs and 27% had detectable eggs. 15% still had detectable living eggs after 1 course of treatment. At follow-up it was demonstrated that antibodies continued to increase for up to 6 months after treatment and average duration of positive GAA antibody was approximately 10 y. The study confirms that the GAA- and MBA-IFAT are not suitable in monitoring results of therapy. Treatment failure in 15% of non-immune patients indicates that studies are needed to define the correct dose of praziquantel in those individuals or to evaluate if resistance to praziquantel is a growing problem.

  9. Comparative Evaluation of Anthelmintic Activity of Edible and Ornamental Pomegranate Ethanolic Extracts against Schistosoma mansoni

    Directory of Open Access Journals (Sweden)

    Doaa A. Yones

    2016-01-01

    Full Text Available Due to the development of praziquantel (PZQ schistosomes resistant strains, the discovery of new antischistosomal agents is of high priority in research. This work reported the in vitro and in vivo effects of the edible and ornamental pomegranate extracts against Schistosoma mansoni. Leaves and stem bark ethanolic extracts of both dried pomegranates were prepared at 100, 300, and 500 μg/mL for in vitro and 600 and 800 mg/kg for in vivo. Adult worms Schistosoma mansoni in RPMI-1640 medium for in vitro and S. mansoni infected mice for in vivo tests were obtained from Theodor Bilharz Research Institute, Cairo, Egypt. In vitro activity was manifested by significant coupled worms separation, reduction of motor activity, lethality, and ultrastructural tegumental alterations in adult worms. In vivo activity was manifested revealed by significant reduction of hepatic granulomas number and diameter, decreased number of bilharzial eggs in liver tissues, lowered liver inflammatory infiltration, decreased hepatic fibrosis, and inducible nitric oxide synthase (iNOS expression. Ethanolic stem bark extract of edible pomegranate exhibited highest antischistosomal activities both in vitro and in vivo. Therefore, pomegranate showed a good potential to be used as a promising new candidate for the development of new schistosomicidal agents.

  10. Spatially explicit Schistosoma infection risk in eastern Africa using Bayesian geostatistical modelling

    DEFF Research Database (Denmark)

    Schur, Nadine; Hürlimann, Eveline; Stensgaard, Anna-Sofie

    2013-01-01

    Schistosomiasis remains one of the most prevalent parasitic diseases in the tropics and subtropics, but current statistics are outdated due to demographic and ecological transformations and ongoing control efforts. Reliable risk estimates are important to plan and evaluate interventions in a spat......Schistosomiasis remains one of the most prevalent parasitic diseases in the tropics and subtropics, but current statistics are outdated due to demographic and ecological transformations and ongoing control efforts. Reliable risk estimates are important to plan and evaluate interventions...... in a spatially explicit and cost-effective manner. We analysed a large ensemble of georeferenced survey data derived from an open-access neglected tropical diseases database to create smooth empirical prevalence maps for Schistosoma mansoni and Schistosoma haematobium for a total of 13 countries of eastern...... Africa. Bayesian geostatistical models based on climatic and other environmental data were used to account for potential spatial clustering in spatially structured exposures. Geostatistical variable selection was employed to reduce the set of covariates. Alignment factors were implemented to combine...

  11. Effects of NaCl Stress on Chlorophyll Fluorescence Parameters in Cinnamomum japonicum var. chenii%NaCl胁迫对普陀樟叶绿素荧光参数的影响

    Institute of Scientific and Technical Information of China (English)

    李影丽; 汪奎宏; 杜国坚; 许利群; 杨华; 肖纪军

    2008-01-01

    [Objective] Study on the changes of chlorophyll fluorescence parameters in Cinnamomum japonicum var. chenii under NaCl stress. [Method] The seedling growth increment, chlorophyll content and chlorophyll fluorescence parameters in leaves of 1-year old Cinnamomum japonicum var. chenii were investigated in field experiment. [Result] Under NaCl stress, seedling growth increment reduced and the chlorophyll content decreased to a stable value; changes of Fv/Fm and Fv/Fo showed identical increasing trend and double peak type. With the aggravation of salt stress, most variations were ob- served in Fo, correlations among chlorophyll fluorescence parameters presented "rise-drop" trend (in the treatment of 7 g/L NaCl). [Condusion] Cinna-momum japonicum var. chenii is endowed with strong salt resistance and wide adaptability.

  12. BIOCHEMICAL AND IMMUNOLOGICAL CHARACTERIZATION OF THE 200kD SCHISTOSOMULUM SURFACE ANTIGEN COMMON TO SCHISTOSOMA MANSONI AND SCHISTOSOMA JAPONICUM%曼氏和日本血吸虫童虫体表200kD共同抗原的生化与免疫学特性

    Institute of Scientific and Technical Information of China (English)

    易新元; 周金春; 王庆林

    2000-01-01

    Objective To study the biochemecal and immunological characterization of the 200 kD schistosomulum surface antigen Method and results A very high molecular weight schistosomulum surface antigen of Mr>200kD was identified and characterized using monoclonal antibodies. Carbohydrate modification experiments followed by radioimmunobinding assays demonstrated that the epitope recognised by the mAbs on this antigen was carbohydrate in nature, while protein digestion experiments followed by SDS-PAGE indicated that this antigen also contained protein. Immunoprecipitation of 125I-labelled cercarial, schistosomulum, adult worm and miracidial surface antigens followed by gel analysis showed the carbohydrate epitope to be present on 5 cercarial, 2 schistosomulum and 5 miracidial surface molecules, and suggested a possible ecological function involved in adapting the parasite to the aquatic free-living stages of its life cycle and possibly also in protecting the early schistosomula from host immune damage. The 5 cercarial surfacs antigens proved to be associated with the CHR, since all the mAbs which recognised those antigens could induce a strong CHR. A kinetic investigation of the carbohydrate epitope on schistosomula of different ages demonstrated a gradual and possibly irreversible loss during the culture period. The epitope completely disappeared from the surface of adult worms. Conclusion To demonstrate an epitope common to a number of surface molecules of various developmental stages of schistosome and therefore explains the immunological cross - reactivity among different stages at the molecular level.

  13. Sero-prevalence of Schistosoma species in cattle in Maiduguri Metropolis and Jere Local Government Areas of Borno State, Nigeria

    Directory of Open Access Journals (Sweden)

    Idris Umar Hambali

    2016-03-01

    Full Text Available Objectives: This study was designed to investigate the Sero-prevalence of Schistosoma species in cattle in Maiduguri Metropolis (MMC and Jere Local Government Areas (LGAs of Borno State, Nigeria. Materials and Method: Blood samples (n=200 from cattle were collected using a multistage sampling technique; 100 samples each were collected from MMC and Jere LGAs, respectively. The samples were subjected to screening for Schistosoma antibodies using Enzyme Linked Immunosorbent Assay (ELISA. Age, sex, breed and location of cattle were recorded. Results: The overall prevalence of Schistosoma infection among cattle in MMC and Jere LGAs was 10%. Jere LGA had a prevalence rate of 14% and MMC had 6%. At the ward levels, Custom Area in Jere LGA had the highest number of Schistosoma positive (50%. Out of 103 female and 97 male cattle screened, the prevalence of Schistosoma infection in female and male were 9.71% (n=10/103 and 10.31% (n=10/103. Out of the 177 serum samples from cattle aging >1-year (adult examined, 16 (9.04% were positive, while only 4 (17.39% out of 23 serum samples from cattle aging <1-year (young were positive. Out of the eight (8 breeds screened, the highest number of cases was recorded in Kuri breed (16.22%. This was followed by Sokoto Gudali (10.9% breed. The prevalence in other breeds was as follows: Abore- 10%, Red Bororo- 5.26%, and White Fulani- 6.67%. Conclusion: It is concluded that schistosomiasis in cattle was prevalent in MMC and Jere LGAs of Borno State. A regular checking program is suggested to constantly check out whether the prevalence rate is increasing, so that effective control measures can be strenthened. [J Adv Vet Anim Res 2016; 3(1.000: 56-61

  14. Right bundle branch block

    DEFF Research Database (Denmark)

    Bussink, Barbara E; Holst, Anders Gaarsdal; Jespersen, Lasse;

    2013-01-01

    AimsTo determine the prevalence, predictors of newly acquired, and the prognostic value of right bundle branch block (RBBB) and incomplete RBBB (IRBBB) on a resting 12-lead electrocardiogram in men and women from the general population.Methods and resultsWe followed 18 441 participants included.......5%/2.3% in women, P Right bundle branch block was associated with significantly.......60-1.62). The presence of IRBBB was not associated with any adverse outcome.ConclusionIn this cohort study, RBBB and IRBBB were two to three times more common among men than women. Right bundle branch block was associated with increased cardiovascular risk and all-cause mortality, whereas IRBBB was not. Contrary...

  15. Determination of Flavonoids in Different Parts and Different Harvesting Seasons of Hypericum japonicum%不同采收季节及部位田基黄药材中黄酮类成分的含量测定

    Institute of Scientific and Technical Information of China (English)

    席仲洪; 虞金宝; 吕武清; 李晶; 李才堂

    2009-01-01

    目的:测定不同采收季节及部位田基黄药材中黄酮类成分的含量.方法:采用比色法测定田基黄药材中总黄酮的含量;采用高效液相色谱(HPLC)法测定田基黄药材中槲皮素、芦丁、异鼠李素的含量.结果:不同采收季节田基黄药材中总黄酮的含量无显著差异;广西产田基黄药材中不含异鼠李素,但槲皮素、芦丁含量较高.田基黄药材不同部位中总黄酮含量以种子和叶高,根中低,且根中几乎不含异鼠李素、槲皮素、芦丁.结论:由于春季田基黄药材植株矮小、产量低,因此应以秋季采收为佳.规定田基黄药材的药用部位为全草是科学的.%OBJECTIVE: To determine the contents of flavonoids in different part and different harvesting seasons of Hypericum japonicum. METHODS: The total content of flavonoids in H. japonicum was determined by colorimetry and the contents of quercetin, rutin and isorhamnetin in H, japonicum were determined by HPLC. RESULTS: The H. japonicum harvested in different seasons showed no significant difference in terms of the contents of total flavonoids, H. japonicum from Guangxi province contained no isorhamnetin but which was rich in the contents of quercetin and rutin. In H. japonicum, the content of total flavonoids was rich in the seeds and leaves but poor in the roots, and hardly had isorhamnetin, quercetin and rutin been detected in the roots. CONCLUSION: H. japonicum harvested in the spring season is low and small in size with low - yield; therefore, fall is its optimal harvesting season. Furthermore, the specification in Chinese Pharmacopeia that the medicinal part of H. japonicum is the entire herb has been proved to be scientific.

  16. Contaminated soil concrete blocks

    NARCIS (Netherlands)

    Korte, de A.C.J.; Brouwers, H.J.H.; Limbachiya, Mukesh C.; Kew, Hsein Y.

    2009-01-01

    According to Dutch law the contaminated soil needs to be remediated or immobilised. The main focus in this article is the design of concrete blocks, containing contaminated soil, that are suitable for large production, financial feasible and meets all technical and environmental requirements. In ord

  17. Effects of Block Scheduling

    Directory of Open Access Journals (Sweden)

    William R. Veal

    1999-09-01

    Full Text Available This study examined the effects of a tri-schedule on the academic achievement of students in a high school. The tri-schedule consists of traditional, 4x4 block, and hybrid schedules running at the same time in the same high school. Effectiveness of the schedules was determined from the state mandated test of basic skills in reading, language, and mathematics. Students who were in a particular schedule their freshman year were tested at the beginning of their sophomore year. A statistical ANCOVA test was performed using the schedule types as independent variables and cognitive skill index and GPA as covariates. For reading and language, there was no statistically significant difference in test results. There was a statistical difference mathematics-computation. Block mathematics is an ideal format for obtaining more credits in mathematics, but the block format does little for mathematics achievement and conceptual understanding. The results have content specific implications for schools, administrations, and school boards who are considering block scheduling adoption.

  18. Concrete Block Pavements

    Science.gov (United States)

    1983-03-01

    1967, Cedergren 1974, Federal Highway .’,U .. V,47 -’":: 37 Administration 1980). Block pavements have essentially the same prob- lems with moisture...Vicksburg, Miss. Cedergren , H. R. 1974. Drainage of Highway and Airfield Pavements, John Wiley and Sons, New VOk. I Cement and Concrete Association

  19. Block That Pain!

    Science.gov (United States)

    ... along with the National Institute of Dental and Craniofacial Research (NIDCR) and the National Institute of General Medical Sciences (NIGMS). This finding shows that a specific combination of two molecules can block only pain-related neurons. It holds the promise of major ...

  20. "In vivo" leukocyte chemotaxis in experimental mice Schistosoma mansoni infection Quimiotaxia de leucócitos "in vivo" na infecção experimental por Schistosoma mansoni

    Directory of Open Access Journals (Sweden)

    Kirte Maria Teixeira

    1994-06-01

    Full Text Available The "in vivo" chemotaxis was studied in C57B1/10 mice 10, 30, 50 and 60 days after a Schistosoma mansoni infection in comparison to a control group (uninfected mice. Staphylococcal protein A was injected into a connective tissue air pouch of control and experimental mice and the leukocyte chemotaxis was counted. A decrease in polymorphonuclear (PMN leukocyte response was found in infected mice in comparison to the control group (pA quimiotaxia de leucócitos "in vivo" foi avaliada em camundongos da linhagem C57B1/10 e estudada 10, 30, 50 e 60 dias após a infecção por Schistosoma mansoni. A proteína A foi utilizada como quimiotático e injetada no tecido conjuntivo no dorso dos camundongos dos grupos experimentais e controle. Nos grupos experimentais foi observado uma diminuição na resposta dos leucócitos polimorfonucleares (PMN em comparação com o grupo controle (p<0.05. Os camundongos estudados 10 dias após a infecção, mostraram uma diminuição na resposta quimiotática de leucócitos PMN, comparando com o grupo controle (p<0.05 e este dado tornou-se mais evidente nos grupos experimentais estudados 30 e 50 dias após a infecção. Apesar da resposta quimiotática dos leucócitos PMN nos camundongos estudados 60 dias após a infecção aumentarem em comparação aos animais analisados 50 dias após a infecção, este aumento foi bem menor em relação ao grupo controle. A resposta quimiotática dos mononucleares não apresentou diferença significativa entre camundongos experimentais e controles

  1. Tegumental Ca-stimulated adenosine triphosphatase activity in adult Schistosoma mansoni worms Atividade da adenosina trifosfatase estimulada pelo Ca no tegumento de vermes adultos de Schistosoma mansoni

    Directory of Open Access Journals (Sweden)

    Italo M. Cesari

    1989-09-01

    Full Text Available A Ca-stimulated ATPase activity (pH 9.5 associated with the tegumental membrane enriched (TME fraction of Schistosoma mansoni adults was partially inhibited by NAP-taurine or by increasing concentrations of chlorpromazine; endogenous calmodulin was found associated with the TME fraction. A similar activity (pH 8.6 was histochemically visualized whithin the tegument of fixed worms on the cytoplasmic leaflet of both the doubel surface membrane and the basement membrane; this reaction was inhibited by 1 µM chloropromazine and it was also observed on the inner side of double membrane vesicles present in the TME fraction. No ATPase activity could be seen at alkaline pH with added Mg or Na/K ions. Without ATP, the addition of external Ca to the fixed worms induced the appearance of lead precipitates on the tegumental discoid bodies; this reaction was inhibited by molybdate and not by chlorpromazine. The intrategumentary regulation of calcium by the systems described and the possible use of phenothiazines against schistosimes are discussed.A atividade ATPse (pH 9.5 estimulada por ions de Ca associados a uma fração enriquecida de membranas do tegumento (fração EMT de vermes adultos de Schistosoma mansoni, foi inibida pro NAP-taurina ou por concentrações crescentes de clorpromacina. Foi encontrada calmodulina enfogena associada principlamente a esta fração. Em vermes adultos fixados com glutaraldeido se detectou histoquimicamente uma atividade ATPase similar (pH 8.6 na face citoplasmática da dupla membrana de superfície e da membrana por 1 µM de clorpromacina e foi também observada na face interna de vesículas de dupla membrana presentes na fração EMT. Não se pôde detectar atividade ATpase em pH alcalino na presença de ions de Mg ou Na/K. A adição externa de Ca, sem ATP, aos vermes fixados induz ao aparecimento de precipitados nos corpos discóides do tegumento; esta reação foi inibida. Os resultados são discutidos em relação a

  2. Oogram studies in mice infected with Schistosoma mansoni and treated with dexamethasone Oograma em camundongos infectados com Schistosoma mansoni e tratados com dexametasona

    Directory of Open Access Journals (Sweden)

    Marco Victor Hermeto

    1994-04-01

    Full Text Available Mice infected with about 90 cercariae of Schistosoma mansoni (LE strain were treated during five consecutive days with dexamethasone (50 mg/Kg, subcutaneously, starting on the 42th day of infection. Groups of five mice were then daily sacrificed from the first day after onset of treatment until the first day after. The perfusion of the portal system was performed and a piece of the intestine was processed for qualitative and quantitative oograms. This treatment carries to larger numbers of eggs in the tissues of treated mice, when compared with untreated groups. No changes were observed in the kinetics of oviposition, as all stages of viable eggs were observed in the tissues of treated and control mice. These data reinforce the hypothesis of a partial blockade of the egg excretion in immunossupressed mice.Camundongos infectados com cerca de 90 cercárias da cepa LE de Schistosoma mansoni foram tratados durante 5 dias consecutivos com dexametasona (50mg/ Kg, subcutaneamente a partir do 42º dia de infecção. Grupos de cinco camundongos foram sacrificados diariamente após o primeiro dia do início do tratamento até o primeiro dia após o término. A perfusão do sistema porta foi feita e fragmentos do intestino foram processados para a realização de oogramas qualitativos e quantitativos. O tratamento leva a um maior número de ovos nos tecidos dos camundongos tratados, se comparado com os grupos não tratados. Nenhuma mudança foi observada na cinética de oviposição, e ovos viáveis em todos os estádios evolutivos foram observados nos tecidos de camundongos tratados e controles. Estes dados reforçam a hipótese de um bloqueio parcial na saída de ovos dos tecidos do intestino para o lúmem intestinal em camundongos imunossuprimidos.

  3. Isolation and specific detection of two major schistosoma gut-associated circulating antigens%血吸虫肠相关循环抗原组分CAA和CCA的纯化与特异检测

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    目的为探讨血吸虫肠相关循环抗原CAA与CCA诊断靶微粒上表位特异性的差别,并试探获取其纯化制品用于定量检测的标准系列。 方法对两组分进行了亲和层析及阴离子交换剂高压液相纯化分离,并应用单抗检测系统进行同相和异相交互测试。 结果 Mono-Q HPLC梯度洗脱分离AWAj-TCA可溶组分,可获得一个带阳离子活性的非结合CCA-1峰及3个大小不等的,带阳离子活性的CCA-2,CCA-3,CCA-4非结合洗脱峰,以及一个带阴离子活性CAA-1洗脱峰。CAA活性峰在峰谱上与CCA-3有部分重叠。与单抗亲和层析纯品的活性对比测定显示CCA-1与CCA-2为该组分的主要构成,但CCA-2及CAA-1在本实验条件下均有微量的相互杂染。同相和异相双位点ELISA的4种组合交互检测,展示CCA只能在捕获与检测抗体同为抗CCA单抗的一种组合中被检示,而另3种组合都只能测出CAA组分。 结论血吸虫肠相关CCA组分为一兼含两性电荷的分子混合体;而CAA分子上具有一个可被抗CCA单抗识别的活性表位位点,从而可能影响纯化分离和特异检测。%Objectives To investigate the nature of the common epitopes of Schistosoma japonicum (S. japonicum) circulating anodic (CAA) and circulating cathodic antigen (CCA) and to try to obtain sufficient purified material to set up a standard series for quantitative determinations. Methods Isolation of the two worm fractions from a trichloroacetic acid (TCA) soluble preparation of S. japonicum adult worm antigen (AWAj-TCA) via Mono-Q anion exchange chromatography was performed and analysis of specific reactivity of the eluted fractions was done by antigen-capture Enzyme Linked Immuno Sorbent Assay (ELISA) specific for CAA or CCA with reference to affinity purified preparations of S. mansoni CAA and CCA. Results When an ionic strength gradient was used, CCA was eluted in two major peaks, an unbound

  4. Utility of Schistosoma bovis Adult Worm Antigens for Diagnosis of Human Schistosomiasis by Enzyme-Linked Immunosorbent Assay and Electroimmunotransfer Blot Techniques

    Science.gov (United States)

    Pardo, J.; Carranza, C.; Turrientes, M. C.; Arellano, J. L. Pérez; Vélez, R. López; Ramajo, V.; Muro, A.

    2004-01-01

    Immunodiagnostic methods based on the detection of antibodies continue to be the most effective and practical methods for the diagnosis of imported schistosomiasis. Schistosoma bovis is a species whose final natural hosts are bovines, ovines, caprines, and small wild ruminants. Different studies have demonstrated the analogies existing between S. bovis and other Schistosoma species which affect humans. The objective of this work was to evaluate the utility of S. bovis adult worm antigens (AWA) for the diagnosis of imported human schistosomiasis by enzyme-linked immunosorbent assay (ELISA) and electroimmunotransfer blotting (EITB) techniques. By detecting eggs, the ELISA for S. bovis AWA was able to definitively detect imported cases with a sensitivity of 94%. The specificity of the ELISA for S. bovis AWA was 97%. There were no differences between the results of the S. bovis AWA ELISA for patients infected with Schistosoma mansoni and those infected with Schistosoma haematobium. The EITB technique showed bands of 85, 37, and 20 kDa, which are characteristic of infections with Schistosoma spp. Specific bands to indicate infection by different species of Schistosoma have not been detected. The combined use of the ELISA for S. bovis AWA and EITB increased the global sensitivity of the study to 97%. Our findings suggest that the ELISA for S. bovis AWA is a useful test for the immunodiagnosis of imported schistosomiasis and that EITB for detecting S. bovis AWA permits the confirmation of diagnosis when the ELISA for S. bovis AWA is positive. PMID:15539523

  5. Genomic organization and genomic structural rearrangements of Sphingobium japonicum UT26, an archetypal γ-hexachlorocyclohexane-degrading bacterium.

    Science.gov (United States)

    Nagata, Yuji; Natsui, Shunsuke; Endo, Ryo; Ohtsubo, Yoshiyuki; Ichikawa, Natsuko; Ankai, Akiho; Oguchi, Akio; Fukui, Shigehiro; Fujita, Nobuyuki; Tsuda, Masataka

    2011-12-10

    The complete genome sequencing of a γ-hexachlorocyclohexane-degrading strain, Sphingobium japonicum UT26, revealed that the genome consists of two circular chromosomes [with sizes of 3.5 Mb (Chr1) and 682kb (Chr2)], a 191-kb large plasmid (pCHQ1), and two small plasmids with sizes of 32 and 5kb. The lin genes are dispersed on Chr1, Chr2, and pCHQ1. Comparison of the UT26 genome with those of other sphingomonad strains demonstrated that the "specific"lin genes for conversion of γ-HCH to β-ketoadipate (linA, linB, linC, linRED, and linF) are located on the DNA regions unique to the UT26 genome, suggesting the acquisition of these lin genes by horizontal transfer events. On the other hand, linGHIJ and linKLMN are located on the regions conserved in the genomes of sphingomonads, suggesting that the linGHIJ-encoded β-ketoadipate pathway and the LinKLMN-type ABC transporter system are involved in core functions of sphingomonads. Based on these results, we propose a hypothesis that UT26 was created by recruiting the specific lin genes into a strain having core functions of sphingomonads. Most of the specific lin genes in UT26 are associated with IS6100. Our analysis of spontaneous linA-, linC-, and linRED-deletion mutants of UT26 revealed the involvement of IS6100 in their deduced genome rearrangements. These facts strongly suggest that IS6100 plays important roles both in the dissemination of the specific lin genes and in the genome rearrangements.

  6. Recovery from blocking between outcomes.

    Science.gov (United States)

    Wheeler, Daniel S; Miller, Ralph R

    2005-10-01

    Contemporary associative learning research largely focuses on cue competition phenomena that occur when 2 cues are paired with a common outcome. Little research has been conducted to investigate similar phenomena occurring when a single cue is trained with 2 outcomes. Three conditioned lick suppression experiments with rats assessed whether treatments known to alleviate blocking between cues would also attenuate blocking between outcomes. In Experiment 1, conditioned responding recovered from blocking between outcomes when a long retention interval was interposed between training and testing. Experiment 2 obtained recovery from blocking between outcomes when the blocking outcome was extinguished after the blocking treatment. In Experiment 3, a recovery from blocking between outcomes occurred when a reminder stimulus was presented in a novel context prior to testing. Collectively, these studies demonstrate that blocking of outcomes, like blocking of cues, appears to be caused by a deficit in the expression of an acquired association.

  7. SUPERFICIAL CERVICAL PLEXUS BLOCK

    Directory of Open Access Journals (Sweden)

    Komang Mega Puspadisari

    2014-01-01

    Full Text Available Superficial cervical plexus block is one of the regional anesthesia in  neck were limited to thesuperficial fascia. Anesthesia is used to relieve pain caused either during or after the surgery iscompleted. This technique can be done by landmark or with ultrasound guiding. The midpointof posterior border of the Sternocleidomastoid was identified and the prosedure done on thatplace or on the level of cartilage cricoid.

  8. Scanning electron microscopy of the tegumental surface of adult Schistosoma spindale.

    Science.gov (United States)

    Kruatrachue, M; Riengrojpitak, S; Upatham, E S; Sahaphong, S

    1983-09-01

    The tegumental surfaces of adult male and female of Schistosoma spindale were studied by scanning electron microscopy. In general, the body surface of the male appears to be fairly uniform from anterior end to posterior end. It is characterized by the presence of transverse ridges and papillae of various types. These papillae are distributed fairly regularly over the whole body surface of the worm. The tegument lining the gynecophoral canal of the male worm is covered with numerous spines interspersed with papillae, some without cilia and some with crater-like holes in the centres and apical cilia. The tegument of the female worm is covered with smooth and perforated ridges and sensory bulbs with apical nodules.

  9. Acute Appendicitis and Pneumatosis in a Duplicated Appendix With Schistosoma Remnants.

    Science.gov (United States)

    Handra-Luca, Adriana; Bisseret, Damien; Dragoescu, Ema

    2016-02-01

    Appendiceal pneumatosis is rare, reported either in the context of acute appendicitis or enterocolitis. Here, we report the case of an elderly adult in whom the acute appendicitis was associated with pneumatosis and occurred in the context of a malformed appendix with pathogenic organism remnants. A 72-year-old man presented with abdominal pain 3 weeks after posttraumatic dorsolumbar surgery. The computed tomography scan showed acute appendicitis and 2 diverticula. On microscopy, the appendix showed acute appendicitis along with a Cave-Wallbridge type A duplication. In addition, several optically clear spaces were observed in the entire appendiceal wall consistent with pneumatosis of the appendix. Focally, calcified structures suggesting pathogenic organisms such as Schistosoma were noted as well. In conclusion, we report a case of appendiceal pneumatosis occurring in the context of acute appendicitis in a duplicated appendix, with presence of calcified structures suggestive of pathogenic organisms.

  10. Immunostimulatory effects of extract of Pulicaria crispa before and after Schistosoma mansoni infection.

    Science.gov (United States)

    Maghraby, Amany S; Shalaby, Nagwa; Abd-Alla, Howida I; Ahmed, Samia A; Khaled, Hussein M; Bahgat, Mahmoud M

    2010-01-01

    The immunostimulatory effects of methanolic extract from Pulicaria crispa were investigated in mice before and after infection with Schistosoma mansoni. Mice were subjected for daily intra-peritoneal injection by the extract (33 ng/mouse) for 10 successive days followed by infecting every mouse with 100 S. mansoni cercariae. Treatment with the extract induced significant increase (p < 0.05) in sera-IL-2 before and after infection. Upon using soluble worm antigen preparation or cancer bladder homogenates as antigens in ELISA, the detected levels of IgG were significantly (p < 0.05) higher in sera from treated-infected mice than untreated P. crispa infected mice. Using crude Escherichia coli lysate as an antigen in ELISA, it was detected a significant (p < 0.05) increase in IgG levels in sera from the extract-treated mice before and after infection.

  11. Construction and characterization of a Schistosoma mansoni bacterial artificial chromosome library.

    Science.gov (United States)

    Le Paslier, M C; Pierce, R J; Merlin, F; Hirai, H; Wu, W; Williams, D L; Johnston, D; LoVerde, P T; Le Paslier, D

    2000-04-15

    A bacterial artificial chromosome (BAC) library has been established from genomic DNA isolated from the trematode parasite of human, Schistosoma mansoni. This library consists of more than 21,000 recombinant clones carrying inserts in the pBeloBAC11 vector. The mean insert size was 100 kb, representing an approximate 7.95-fold genome coverage. Library screening with eight chromosome-specific or single-copy gene probes yielded between 1 and 9 positive clones, and none of those tested was absent from the library. End sequences were obtained for 93 randomly selected clones, and 37 showed sequence identity to S. mansoni sequences (ESTs, genes, or repetitive sequences). A preliminary analysis by fluorescence in situ hybridization localized 8 clones on schistosome chromosomes 1 (2 clones), 2, 3, 5, Z, and W (3 clones). This library provides a new resource for the physical mapping and sequencing of the genome of this important human pathogen.

  12. Purification and characterization of an elastinolytic proteinase secreted by cercariae of Schistosoma mansoni.

    Science.gov (United States)

    McKerrow, J H; Pino-Heiss, S; Lindquist, R; Werb, Z

    1985-03-25

    An elastinolytic proteinase secreted by tissue-invasive larvae of Schistosoma mansoni has been purified to homogeneity. Size-exclusion chromatography and chromatofocusing were used to purify the enzyme 18-fold from crude larval secretions. The native enzyme has a molecular weight of 30,000, a pI of 8, a pH optimum of 9, and a calcium dependence of 2 mM. A second Mr 17,000 form of the enzyme was present in crude secretions and appears to be an autoproteolysis product. The enzyme is a serine proteinase that preferentially binds tetrapeptide inhibitors or substrates with an aromatic or hydrophobic residue at the P-1 site. In addition to being active against elastin, the enzyme degrades Azocoll, gelatin, laminin, fibronectin, keratin, and type IV collagen.

  13. The growth and development of Schistosoma mansoni in mice exposed to sublethal doses of radiation

    Energy Technology Data Exchange (ETDEWEB)

    Aitken, R.; Wilson, R.A. (Univ. of York, Heslington (England))

    1989-12-01

    The maturation of Schistosoma mansoni was studied in mice exposed to various sublethal doses of radiation. Although the treatment of mice with 500 rads of radiation prior to infection did not alter parasite maturation, doses in excess of 500 rads led to a reduction in worm burden. This could not be attributed to a delay in the arrival of parasites in the hepatic portal system. Worms developing in mice treated with 800 rads commenced egg-laying about 1 wk later than worms in intact mice, and the rate of egg deposition appeared to be lower in irradiated hosts. The data demonstrate that exposure of C57BL/6 mice to doses of radiation in excess of 500 rads impairs their ability to carry infections of S. mansoni. The findings do not support the hypothesis that primary worm burdens in the mouse are controlled by a host immune response.

  14. Development of -omics research in Schistosoma spp. and omics-based new diagnostic tools for schistosomiasis

    Directory of Open Access Journals (Sweden)

    Wei eHu

    2014-06-01

    Full Text Available Schistosomiasis, caused by dioecious flatworms in the genus Schistosoma, is torturing people from many developing countries nowadays and frequently leads to severe morbidity and mortality of the patients. Praziquantel (PZQ based chemotherapy and morbidity control for this disease adopted currently necessitate viable and efficient diagnostic technologies. Fortunately, those -omics researches, which rely on high-throughput experimental technologies to produce massive amounts of informative data, have substantially contributed to the exploitation and innovation of diagnostic tools of schistosomiasis. In its first section, this review provides a concise conclusion on the progresses pertaining to schistosomal omics researches to date, followed by a comprehensive section on the diagnostic methods of schistosomiasis, especially those innovative ones based on the detection of antibodies, antigens, nucleic acids, and metabolites with a focus on those achievements inspired by omics researches. Finally, suggestions about the design of future diagnostic tools of schistosomiasis are proposed, in order to better harness those data produced by omics studies.

  15. Effects of Endosulfan on Predator-Prey Interactions Between Catfish and Schistosoma Host Snails.

    Science.gov (United States)

    Monde, Concillia; Syampungani, Stephen; Van den Brink, Paul J

    2016-08-01

    The effect of the pesticide endosulfan on predator-prey interactions between catfish and Schistosoma host snails was assessed in static tank experiments. Hybrid catfish (Clarias gariepinus × C. ngamensis) and Bulinus globosus were subjected to various endosulfan concentrations including an untreated control. The 48- and 96-h LC50 values for catfish were 1.0 and snails were 1137 and 810 µg/L. To assess sublethal effects on the feeding of the catfish on B. globosus, endosulfan concentrations between 0.03 and 1.0 µg/L were used. Predation was significantly greater (p snails using fish may be affected in endosulfan-polluted aquatic systems of Southern Africa because it has been found present at concentrations that are indicated to cause lethal effects on the evaluated hybrid catfish and to inhibit the predation of snails by this hybrid catfish.

  16. Compatibility of Ugandan Schistosoma mansoni isolates with Biomphalaria snail species from Lake Albert and Lake Victoria

    DEFF Research Database (Denmark)

    Adriko, Moses; Standley, Claire J.; Tinkitina, Benjamin

    2013-01-01

    In order to investigate the capacity of being intermediate host for Schistosoma mansoni, the Ugandan F1 generation of Biomphalaria snail species that were laboratory-bred from parent populations originally collected from either Lake Victoria or Lake Albert was challenged with sympatric and non......-sympatric S. mansoni isolates. After a prepatent period of 20 days, a daily 10-hourly snail shedding for cercariae was done to determine the infection rate, cercarial production per hour and survival period of infected snails. The study suggests that when parasite strains from a different geographical origin...... is used for infection, survival of infected snails increase, leading to an increased transmission potential. Although earlier literature had indicated that the Lake Victoria Biomphalaria sudanica is refractory to S. mansoni, we showed that all Ugandan Biomphalaria spp., including B. sudanica from all...

  17. The schistosoma-specific antibody response after treatment in non-immune travellers

    DEFF Research Database (Denmark)

    Duus, Liv Marie; Christensen, Anders Vittrup; Navntoft, Dorte;

    2009-01-01

    ) and Membrane Bound Antigen (MBA) assays in assessment of treatment efficacy and number of treated non-immune individuals with signs of treatment failure. In a retrospective study, residents in Denmark diagnosed with positive Schistosoma antibodies in the period 1987 - 2004 were offered follow-up including...... analyses for GAA, MBA, IgE and eosinophil count. Among 98 patients with positive antibody at time of diagnosis, 73 were examined for eggs and 27% had detectable eggs. 15% still had detectable living eggs after 1 course of treatment. At follow-up it was demonstrated that antibodies continued to increase...... for up to 6 months after treatment and average duration of positive GAA antibody was approximately 10 y. The study confirms that the GAA- and MBA-IFAT are not suitable in monitoring results of therapy. Treatment failure in 15% of non-immune patients indicates that studies are needed to define the correct...

  18. Garlic attenuates histological and histochemical alterations in livers of Schistosoma mansoni infected mice.

    Science.gov (United States)

    Mahmoud, Y I; Riad, N H; Taha, H

    2016-08-01

    Interest in screening for new anti-schistosomal agents is growing because of increased concerns about resistance to and safety of praziquantel. We investigated the anti-schistosomal action of prophylactic and therapeutic doses of garlic on the histological and histochemical alterations caused by Schistosoma mansoni infection. Livers of infected mice were characterized by granulomas, periportal inflammation and fibrosis, hepatocyte vacuolation, fatty degeneration and necrosis, and hypertrophy and pigmentation of Kupffer cells. Significant depletion of carbohydrates and increased lipid vacuoles also were observed. All garlic regimens caused suppression of granuloma formation and amelioration of histological and histochemical changes; the continuous treatment protocol produced the best results. Garlic appears to be a safe and economical anti-schistosomal adjuvant for attenuating the pathogenicity of schistosomiasis.

  19. Evidentiation of Paramyosin (Sm-97 as a Modulating Antigen on Granulomatous Hypersensitivity to Schistosoma mansoni Eggs

    Directory of Open Access Journals (Sweden)

    Hirsch Cristine

    1997-01-01

    Full Text Available A Schistosoma mansoni adult worm anionic fraction (PIII has previously been shown to protect mice against challenge infection and to reduce pulmonary and hepatic granulomatous hypersensitivity. Serum from PIII-immunized rabbit was used to screen a lgt11 cDNA library from S. mansoni adult worm in order to identify antigens capable of modulating granulomatous hypersensitivity. We obtained four clones with 400 (Sm-III.11, 900 (Sm-III.16, 1100 (Sm-III.10 and 1300 (Sm-III.12 bp of length. All clone-specific antibodies were able to recognize most of the PIII components. The sequence analysis showed that these clones presented high homology with S. mansoni paramyosin (Sm-97. These findings ascribe a new function to this antigen with an important role in modulation of granulomatous hypersensitivity to S. mansoni eggs

  20. Managing access block.

    Science.gov (United States)

    Cameron, Peter; Scown, Paul; Campbell, Donald

    2002-01-01

    There is pessimism regarding the ability of the Acute Health Sector to manage access block for emergency and elective patients. Melbourne Health suffered an acute bed crisis in 2001 resulting in record ambulance diversions and emergency department (ED) delays. We conducted an observational study to reduce access block for emergency patients whilst maintaining elective throughput at Melbourne Health. This involved a clinician-led taskforce using previously proven principles for organisational change to implement 51 actions to improve patient access over a three-month period. The primary outcome measures were ambulance diversion, emergency patients waiting more than 12 hours for an inpatient bed, elective throughput and theatre cancellations. Despite a reduction in multi-day bed numbers all primary objectives were met, ambulance diversion decreased to minimal levels, 12-hour waits decreased by 40% and elective throughput was maintained. Theatre cancellations were also minimised. We conclude that access block can be improved by clinician-led implementation of proven process improvements over a short time frame. The ability to sustain change over the longer term requires further study.

  1. Morfologia e desenvolvimento de Schistosoma mansoni Sambon, 1907 em infecções unissexuais experimentalmente produzidas no camundongo Morphology and development of Schistosoma mansoni Sambon, 1907 in unisexual infections produced experimentally in mice

    Directory of Open Access Journals (Sweden)

    Eliana Maria Zanotti

    1982-04-01

    Full Text Available Estudou-se o desenvolvimento de Schistosoma mansoni em infecções unissexuais no camundongo. Os esquistossomos fêmeos apresentaram-se menos desenvolvidos do que os machos. Houve correlação entre o comprimento dos machos e o número de testículos. Verificou-se que o isolamento sexual é prejudicial aos dois sexos, principalmente à fêmea.The Schistosoma mansoni development in mice submitted to unisexual infections was studied. The single female worms developed less than the single males. There was correlation between the male's length and the number of his tests. It was verified that sexual isolation of the schistosomes is prejudicial to both sexes, mainly for the female.

  2. Therapeutic effect of Arctium lappa in Schistosoma haematobium associated kidney disturbance: biochemical and molecular effects.

    Science.gov (United States)

    Koriem, Khaled M M; Idris, Zulzamri H; Haron, Hasniza F; Omar, Nurulhuda A; Lazain, Halita S

    2016-12-01

    Schistosoma haematobium (S. haematobium) infection has been found to be strongly associated with bladder cancer, which necessitates for discover of a natural new therapeutic agent. The aim of this study was to evaluate the therapeutic effect of Arctium lappa seed extract in S. haematobium associated kidney disturbance. Forty male albino mice were used and divided into four equal groups; group 1 control includes non-infected healthy mice, groups 2, 3 and 4 subcutaneous infected with S. haematobium cercariae. Groups 3 co-treated daily with oral dose of A. lappa seed extract (300 mg/kg, bwt) for 15 days in the same time of S. haematobium infection. Groups 4 post-treated daily for 15 days with oral dose of A. lappa seed extract (300 mg/kg, bwt) after 15 days of S. haematobium infection. The results obtained revealed that S. haematobium significantly decreased kidney weight and serum sodium, potassium and chloride, but increased urinary volume, urinary excretion of sodium, potassium and chloride, serum urea, creatinine and uric acid. Schistosoma haematobium also significantly decreased kidney superoxide dismutase, glutathione peroxidase and reduced glutathione levels while increased kidney lipid peroxidation level. Co- and post-treatment with A. lappa seed extract restore all the above parameters to approach the normal values. These results were supported with histopathological examinations. In conclusion, A. lappa seed extract has therapeutic effect in kidney disturbance caused by S. haematobium where co-treatment of A. lappa seed extract was more effective than post-treatment of the extract.

  3. Dynamic Observation of Liver Fibrosis in Mice Infected with Schistosoma japonica

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    The expression of TNF-α in the liver at different periods post Schistosoma japonica infection and the effect on liver fibrosis after supplementary injection of these eytokines were investigated. The mice infected with schistosome cercariae were divided into 3 groups: normal control group, TNF-α-untreated infection group and TNF-α-treated infection group. ABC immunohistochemistry and pathologic image multimedia quantification system were applied to dynamically detect the activity of TNF-α. The results showed that the levels of TNF-α in the liver in TNF-α-untreated infection group were slowly decreased with prolongation of infection time (from 8th, 11th, 14th to 18th week), while in the TNF-α-treated infection group, those were increased significantly after intraperitoneal injection of TNF-α at 6th week after infection. At first to 8th week after the final injection of TNF-α, the intrahepatic TNF-α levels in the TNF-α-treated infection group were significantly higher than in the other two groups (P<0.01), and the granulomatous inflammation and fibrosis in the liver were also milder in the normal control group. It was concluded that at the early stage of Schistosoma japonica infection mouse liver mainly released Th1 cytokine and TNF-α from Th1 activated macrophages. Six weeks after infection (post egg deposition), exogenous supplement with intraperitoneal injection of TNF-α could induce the enhanced expression of Th1 cytokines and alleviate the liver granulomatous inflammation and fibrosis.

  4. Block Transfer Handbook: Constructing and Negotiating Block Transfer Agreements.

    Science.gov (United States)

    Finlay, Finola

    The purpose of this handbook is to provide resources for institutions or articulation committees who are engaged in the task of investigating the feasibility of block transfer agreements. Block transfer is the process whereby a block of credits is granted to students who have successfully completed a certificate, diploma, or cluster of courses…

  5. The New Record of Trichogramma japonicum Ashmead,an Egg Parasitoid of Chilo suppressalis(Walker)in Jilin,China%吉林省稻螟赤眼蜂的发现和鉴定

    Institute of Scientific and Technical Information of China (English)

    郭震; 阮长春; 臧连生; 张帆; 靳锋云

    2011-01-01

    采用二化螟Chilo suppressalis(Walker)卵田间诱集的方法获得赤眼蜂Trichogramma寄生卵,室内培养羽化后,解剖并观察其雄性外生殖器特征,同时进行rDNA-ITS2序列分析,确定其为稻螟赤眼蜂T.japonicum Ashmead.BLAST分析结果表明,当地采集的供试赤眼蜂ITS2与广东的稻螟赤眼蜂相似性高达99.3%~100%.本研究首次确证吉林省存在寄生水稻二化螟的稻螟赤眼蜂自然种群.%Trichogramma wasps were collected using sentinel eggs of Chilo suppressalis (Walker) in rice fields. Throughmorphological identification of male genitalia and analysis of rDNA-ITS2 sequences, a Trichogramma species collectedwas identified as T. Japonicum Ashmead. The results of sequence alignment of ITS2 region showed that Trichogrammasp. Collected in Jilin had 99.3% - 100% similarity with T. Japonicum originated from Guangdong. The present workdemonstrated for the first time that there are natural populations of T. Japonicum parasitizing C. Suppressalis in Jilin,China.

  6. Effect of metal stress on photosynthetic pigments in the Cu-hyperaccumulating lichens Cladonia humilis and Stereocaulon japonicum growing in Cu-polluted sites in Japan.

    Science.gov (United States)

    Nakajima, Hiromitsu; Yamamoto, Yoshikazu; Yoshitani, Azusa; Itoh, Kiminori

    2013-11-01

    To understand the ecology and physiology of metal-accumulating lichens growing in Cu-polluted sites, we investigated lichens near temple and shrine buildings with Cu roofs in Japan and found that Stereocaulon japonicum Th. Fr. and Cladonia humilis (With.) J. R. Laundon grow in Cu-polluted sites. Metal concentrations in the lichen samples collected at some of these sites were determined by inductively coupled plasma mass spectroscopy (ICP-MS). UV-vis absorption spectra of pigments extracted from the lichen samples were measured, and the pigment concentrations were estimated from the spectral data using equations from the literature. Secondary metabolites extracted from the lichen samples were analyzed by high-performance liquid chromatography (HPLC) with a photodiode array detector. We found that S. japonicum and C. humilis are Cu-hyperaccumulating lichens. Differences in pigment concentrations and their absorption spectra were observed between the Cu-polluted and control samples of the 2 lichens. However, no correlation was found between Cu and pigment concentrations. We observed a positive correlation between Al and Fe concentrations and unexpectedly found high negative correlations between Al and pigment concentrations. This suggests that Al stress reduces pigment concentrations. The concentrations of secondary metabolites in C. humilis growing in the Cu-polluted sites agreed with those in C. humilis growing in the control sites. This indicates that the metabolite concentrations are independent of Cu stress.

  7. Effect of praziquantel treatment of Schistosoma mansoni during pregnancy on intensity of infection and antibody responses to schistosome antigens

    DEFF Research Database (Denmark)

    Tweyongyere, Robert; Mawa, Patrice A.; Emojong, Nicholas O.;

    2009-01-01

    Schistosoma mansoni during pregnancy, compared with treatment after delivery. Methods A nested cohort of 387 Schistosoma mansoni infected women was recruited within a larger trial of de-worming during pregnancy. Women were randomised to receive praziquantel or placebo during pregnancy. All women were treated...... after delivery. Infection intensity after treatment was assessed by a single Kato-Katz examination of stool samples with duplicate slides and categorised as undetected, light (1-99 eggs per gram (epg)), moderate (100-399 epg) or heavy (=400 epg). Antibodies against S. mansoni worm and egg antigens were...... infection (median (IQR) epg: 749 (521, 1169)) with S. mansoni. At six weeks after praziquantel treatment during pregnancy S. mansoni infection was not detectable in 81.9% of the women and prevalence and intensity had decreased to 11.8% light, 4.7% moderate and 1.6% heavy a similar reduction when compared...

  8. Dicty_cDB: SLF122 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 08990 Schistosoma mansoni, Phil LoVerde/Joe Merrick Schistosoma mansoni cDNA clon...osoma mansoni, Phil LoVerde/Joe Merrick Schistosoma mansoni cDNA clone SMNCH20 5' end similar to ATPase vacu...histosoma mansoni, Phil LoVerde/Joe Merrick Schistosoma mansoni cDNA clone SMNCG92 5' end similar to ATPase ...SJEBZB10 SJE Schistosoma japonicum cDNA, mRNA sequence. 54 0.003 1 AI067329 |AI067329.1 EST209007 Schistosom...a mansoni, Phil LoVerde/Joe Merrick Schistosoma mansoni cDNA clone SMNAS63 5' end

  9. Schistosoma mansoni venom allergen-like protein 4 (SmVAL4) is a novel lipid-binding SCP/TAPS protein that lacks the prototypical CAP motifs

    Energy Technology Data Exchange (ETDEWEB)

    Kelleher, Alan [Baylor College of Medicine, Houston, TX 77030 (United States); Darwiche, Rabih [University of Fribourg, Chemin du Musée 10, CH 1700 Fribourg (Switzerland); Rezende, Wanderson C. [Baylor College of Medicine, Houston, TX 77030 (United States); Farias, Leonardo P.; Leite, Luciana C. C. [Instituto Butantan, São Paulo, SP (Brazil); Schneiter, Roger [University of Fribourg, Chemin du Musée 10, CH 1700 Fribourg (Switzerland); Asojo, Oluwatoyin A., E-mail: asojo@bcm.edu [Baylor College of Medicine, Houston, TX 77030 (United States)

    2014-08-01

    The first structure of an S. mansoni venom allergen-like protein is presented. Schistosomiasis is a parasitic disease that affects over 200 million people. Vaccine candidates have been identified, including Schistosoma mansoni venom allergen-like proteins (SmVALs) from the SCP/TAPS (sperm-coating protein/Tpx/antigen 5/pathogenesis related-1/Sc7) superfamily. The first SmVAL structure, SmVAL4, was refined to a resolution limit of 2.16 Å. SmVAL4 has a unique structure that could not be predicted from homologous structures, with longer loops and an unusual C-terminal extension. SmVAL4 has the characteristic α/β-sandwich and central SCP/TAPS cavity. Furthermore, SmVAL4 has only one of the signature CAP cavity tetrad amino-acid residues and is missing the histidines that coordinate divalent cations such as Zn{sup 2+} in other SCP/TAPS proteins. SmVAL4 has a cavity between α-helices 1 and 4 that was observed to bind lipids in tablysin-15, suggesting the ability to bind lipids. Subsequently, SmVAL4 was shown to bind cholesterol in vitro. Additionally, SmVAL4 was shown to complement the in vivo sterol-export phenotype of yeast mutants lacking their endogenous CAP proteins. Expression of SmVAL4 in yeast cells lacking endogenous CAP function restores the block in sterol export. These studies suggest an evolutionarily conserved lipid-binding function shared by CAP proteins such as SmVAL4 and yeast CAP proteins such as Pry1.

  10. Nitric oxide production by Biomphalaria glabrata haemocytes: effects of Schistosoma mansoni ESPs and regulation through the extracellular signal-regulated kinase pathway

    Directory of Open Access Journals (Sweden)

    Kirk Ruth S

    2009-04-01

    Full Text Available Abstract Background Schistosoma mansoni uses Biomphalaria glabrata as an intermediate host during its complex life cycle. In the snail, the parasite initially transforms from a miracidium into a mother sporocyst and during this process excretory-secretory products (ESPs are released. Nitric oxide (NO and its reactive intermediates play an important role in host defence responses against pathogens. This study therefore aimed to determine the effects of S. mansoni ESPs on NO production in defence cells (haemocytes from schistosome-susceptible and schistosome-resistant B. glabrata strains. As S. mansoni ESPs have previously been shown to inhibit extracellular signal-regulated kinase (ERK phosphorylation (activation in haemocytes from susceptible, but not resistant, B. glabrata the regulation of NO output by ERK in these cells was also investigated. Results Haemocytes from resistant snails challenged with S. mansoni ESPs (20 μg/ml over 5 h displayed an increase in NO production that was 3.3 times greater than that observed for unchallenged haemocytes; lower concentrations of ESPs (0.1–10 μg/ml did not significantly increase NO output. In contrast, haemocytes from susceptible snails showed no significant change in NO output following challenge with ESPs at any concentration used (0.1–20 μg/ml. Western blotting revealed that U0126 (1 μM or 10 μM blocked the phosphorylation (activation status of ERK in haemocytes from both snail strains. Inhibition of ERK signalling by U0126 attenuated considerably intracellular NO production in haemocytes from both susceptible and resistant B. glabrata strains, identifying ERK as a key regulator of NO output in these cells. Conclusion S. mansoni ESPs differentially influence intracellular NO levels in susceptible and resistant B. glabrata haemocytes, possibly through modulation of the ERK signalling pathway. Such effects might facilitate survival of S. mansoni in its intermediate host.

  11. SmSak, the second Polo-like kinase of the helminth parasite Schistosoma mansoni: conserved and unexpected roles in meiosis.

    Directory of Open Access Journals (Sweden)

    Thavy Long

    Full Text Available Polo-like kinases (Plks are a family of conserved regulators of a variety of events throughout the cell cycle, expanded from one Plk in yeast to five Plks in mammals (Plk1-5. Plk1 is the best characterized member of the Plk family, homolog to the founding member Polo of Drosophila, and plays a major role in cell cycle progression by triggering G2/M transition. Plk4/Sak (for Snk (Serum-inducible kinase akin kinase is a unique member of the family, structurally distinct from other Plk members, with essential functions in centriole duplication. The genome of the trematode parasite Schistosoma mansoni contains only two Plk genes encoding SmPlk1 and SmSak. SmPlk1 has been shown already to be required for gametogenesis and parasite reproduction. In this work, in situ hybridization indicated that the structurally conserved Plk4 protein, SmSak, was largely expressed in schistosome female ovary and vitellarium. Expression of SmSak in Xenopus oocytes confirmed its Plk4 conserved function in centriole amplification. Moreover, analysis of the function of SmSak in meiosis progression of G2-blocked Xenopus oocytes indicated that, in contrast to SmPlk1, SmSak cannot induce G2/M transition in the absence of endogenous Plk1 (Plx1. Unexpectedly, meiosis progression was spontaneously observed in Plx1-depleted oocytes co-expressing SmSak and SmPlk1. Molecular interaction between SmSak and SmPlk1 was confirmed by co-immunoprecipitation of both proteins. These data indicate that Plk1 and Plk4 proteins have the potential to interact and cross-activate in cells, thus attributing for the first time a potential role of Plk4 proteins in meiosis/mitosis entry. This unexpected role of SmSak in meiosis could be relevant to further consider the function of this novel Plk in schistosome reproduction.

  12. Demographic Data - MDC_Block

    Data.gov (United States)

    NSGIC GIS Inventory (aka Ramona) — A polygon feature class of Miami-Dade Census 2000 Blocks. Census blocks are areas bounded on all sides by visible and/or invisible features shown on a map prepared...

  13. Habitat Blocks and Wildlife Corridors

    Data.gov (United States)

    Vermont Center for Geographic Information — Habitat blocks are areas of contiguous forest and other natural habitats that are unfragmented by roads, development, or agriculture. Vermonts habitat blocks are...

  14. Differential anti-glycan antibody responses in Schistosoma mansoni-infected children and adults studied by shotgun glycan microarray.

    Directory of Open Access Journals (Sweden)

    Angela van Diepen

    Full Text Available BACKGROUND: Schistosomiasis (bilharzia is a chronic and potentially deadly parasitic disease that affects millions of people in (subtropical areas. An important partial immunity to Schistosoma infections does develop in disease endemic areas, but this takes many years of exposure and maturation of the immune system. Therefore, children are far more susceptible to re-infection after treatment than older children and adults. This age-dependent immunity or susceptibility to re-infection has been shown to be associated with specific antibody and T cell responses. Many antibodies generated during Schistosoma infection are directed against the numerous glycans expressed by Schistosoma. The nature of glycan epitopes recognized by antibodies in natural schistosomiasis infection serum is largely unknown. METHODOLOGY/PRINCIPAL FINDINGS: The binding of serum antibodies to glycans can be analyzed efficiently and quantitatively using glycan microarray approaches. Very small amounts of a large number of glycans are presented on a solid surface allowing binding properties of various glycan binding proteins to be tested. We have generated a so-called shotgun glycan microarray containing natural N-glycan and lipid-glycan fractions derived from 4 different life stages of S. mansoni and applied this array to the analysis of IgG and IgM antibodies in sera from children and adults living in an endemic area. This resulted in the identification of differential glycan recognition profiles characteristic for the two different age groups, possibly reflecting differences in age or differences in length of exposure or infection. CONCLUSIONS/SIGNIFICANCE: Using the shotgun glycan microarray approach to study antibody response profiles against schistosome-derived glycan elements, we have defined groups of infected individuals as well as glycan element clusters to which antibody responses are directed in S. mansoni infections. These findings are significant for further

  15. Suppression of Basophil Histamine Release and Other IgE-dependent Responses in Childhood Schistosoma mansoni/hookworm Coinfection

    OpenAIRE

    Pinot de Moira, Angela; Fitzsimmons, Colin M.; Frances M Jones; Wilson, Shona; Cahen, Pierre; Tukahebwa, Edridah; Mpairwe, Harriet; Mwatha, Joseph K; Bethony, Jeffrey M.; Skov, Per S.; Kabatereine, Narcis B.; Dunne, David W.

    2014-01-01

    Background.  The poor correlation between allergen-specific immunoglobulin E (asIgE) and clinical signs of allergy in helminth infected populations suggests that helminth infections could protect against allergy by uncoupling asIgE from its effector mechanisms. We investigated this hypothesis in Ugandan schoolchildren coinfected with Schistosoma mansoni and hookworm. Methods.  Skin prick test (SPT) sensitivity to house dust mite allergen (HDM) and current wheeze were assessed pre-anthelmintic...

  16. Porous block nanofiber composite filters

    Energy Technology Data Exchange (ETDEWEB)

    Ginley, David S.; Curtis, Calvin J.; Miedaner, Alexander; Weiss, Alan J.; Paddock, Arnold

    2016-08-09

    Porous block nano-fiber composite (110), a filtration system (10) and methods of using the same are disclosed. An exemplary porous block nano-fiber composite (110) includes a porous block (100) having one or more pores (200). The porous block nano-fiber composite (110) also includes a plurality of inorganic nano-fibers (211) formed within at least one of the pores (200).

  17. Properties of blocked linear systems.

    Science.gov (United States)

    Chen, Weitian; Anderson, Brian D O; Deistler, Manfred; Filler, Alexander

    2012-10-01

    This paper presents a systematic study on the properties of blocked linear systems that have resulted from blocking discrete-time linear time invariant systems. The main idea is to explore the relationship between the blocked and the unblocked systems. Existing results are reviewed and a number of important new results are derived. Focus is given particularly on the zero properties of the blocked system as no such study has been found in the literature.

  18. Appropriate schemata and building blocks

    Institute of Scientific and Technical Information of China (English)

    Yang Haijun; Li Minqiang

    2005-01-01

    Appropriate schemata as a novel concept to characterize building blocks are introduced, and then, the traits of appropriate schemata are presented. The effects of building blocks by search operators are analyzed. Hence, the experiments on RR-8X8 are employed to verify that appropriate schemata construct the building blocks. The validity of appropriate schemata and building blocks from the views of theory and practice is presented.

  19. Blocking the Hawking radiation

    DEFF Research Database (Denmark)

    Autzen, M.; Kouvaris, C.

    2014-01-01

    grows after its formation (and eventually destroys the star) instead of evaporating. The fate of the black hole is dictated by the two opposite mechanics, i.e., accretion of nuclear matter from the center of the star and Hawking radiation that tends to decrease the mass of the black hole. We study how...... the assumptions for the accretion rate can in fact affect the critical mass beyond which a black hole always grows. We also study to what extent degenerate nuclear matter can impede Hawking radiation due to the fact that emitted particles can be Pauli blocked at the core of the star....

  20. Property Blocks: Games and Activities.

    Science.gov (United States)

    Humphreys, Alan, Ed.; Dailey, Jean, Ed.

    This pamphlet describes the property blocks produced by MINNEMAST, and discusses their use in the development of thinking processes. Classification systems, including block diagrams and tree diagrams, are discussed. Sixteen classroom activities and eleven games which use the blocks are described. Suggestions to the teacher for further reading are…

  1. An atlas for Schistosoma mansoni organs and life-cycle stages using cell type-specific markers and confocal microscopy.

    Directory of Open Access Journals (Sweden)

    James J Collins

    Full Text Available Schistosomiasis (bilharzia is a tropical disease caused by trematode parasites (Schistosoma that affects hundreds of millions of people in the developing world. Currently only a single drug (praziquantel is available to treat this disease, highlighting the importance of developing new techniques to study Schistosoma. While molecular advances, including RNA interference and the availability of complete genome sequences for two Schistosoma species, will help to revolutionize studies of these animals, an array of tools for visualizing the consequences of experimental perturbations on tissue integrity and development needs to be made widely available. To this end, we screened a battery of commercially available stains, antibodies and fluorescently labeled lectins, many of which have not been described previously for analyzing schistosomes, for their ability to label various cell and tissue types in the cercarial stage of S. mansoni. This analysis uncovered more than 20 new markers that label most cercarial tissues, including the tegument, the musculature, the protonephridia, the secretory system and the nervous system. Using these markers we present a high-resolution visual depiction of cercarial anatomy. Examining the effectiveness of a subset of these markers in S. mansoni adults and miracidia, we demonstrate the value of these tools for labeling tissues in a variety of life-cycle stages. The methodologies described here will facilitate functional analyses aimed at understanding fundamental biological processes in these parasites.

  2. Development of Schistosoma mansoni in Biomphalaria tenagophila, Biomphalaria straminea and Biomphalaria glabrata Desenvolvimento do Schistosoma mansoni em Biomphalaria tenagophila, Biomphalaria straminea e Biomphalaria glabrata

    Directory of Open Access Journals (Sweden)

    Cecilia Pereira de Souza

    1995-06-01

    Full Text Available A comparative study of the development of Schistosoma mansoni during the intra-molluscan phase was made by means of histological sections of Biomphalaria tenagophila, B. straminea and B. glabrata from Brazil. Two hundred snails of each species were individually exposed to 50 miracidia of the S. mansoni, AL line. No larvae were observed in the snails fixed 72 h after exposure. In specimens shedding cercariae, 31 days after exposure tissue reactions encapsulating the larvae were seen in B. tenagophila and B. straminea, in the head-foot, mantle collar and renal ducts. No tissue reactions occurred in the digestive glands of these two species. In B. glabrata the presence of numerous sporocysts and cercariae without tissue reactions was observed in the digestive gland, and other organs. The levels of infection of the snails and the average numbers of cercariae shed per day were 32.6% and 79±90 respectively for B. tenagophila, 11.3% and 112±100 for B. straminea and 75.3% and 432±436 for B. glabrata. The lower levels of infection and average numbers of cercariae shed by B. tenagophila and B. straminea are thus related to their more potent internal defense systems.Foi feito estudo comparativo do desenvolvimento do Schistosoma mansoni na fase intra-molusco, através de cortes histológicos, em Biomphalaria tenagophila, B. straminea e B. glabrata. Duzentos moluscos de cada espécie foram expostos individualmente a 50 miracídios de S. mansoni da linhagem AL. Nenhuma larva foi observada nos exemplares fixados 72 horas após a exposição. Nos exemplares eliminando cercárías, 31 dias após a exposição, foram observadas reações teciduais de encapsulamento de larvas em B. tenagophila e B. straminea, na região cefalopodal, colar do manto e dutos renais. Nas glândulas digestivas das duas espécies não foram observadas reações. Em B. glabrata foi registrada a presença de numerosos esporocistos e cercárias sem reação tecidual na gl

  3. Portal Doppler ultrasound evaluation in myelitis by Schistosoma mansoni Avaliação pela ultrassonografia com Doppler portal na mielite pelo Schistosoma mansoni

    Directory of Open Access Journals (Sweden)

    Claudio Henrique Fernandes Vidal

    2010-02-01

    Full Text Available The mechanism for development of schistosomal myelitis has not yet been completely understood. Few publications have tried to establish a link between the general clinical presentation of the illness and its neurological manifestations. Objective: To evaluated the portal blood flow in patients with schistosomal myelitis and controls. Method: Forty-three individuals were evaluated using portal Doppler ultrasound. Group I consisted of 13 patients with the spinal form of schistosomiasis. Group II included 10 patients with intestinal and hepatointestinal forms of schistosomiasis, but no neurological symptoms; while group III involved 20 healthy controls. Results: Portal venous speed and portal venous flow were significantly elevated in group I (pA fisiopatogênese da mielopatia esquistossomótica não é completamente entendida, sendo escassas as publicações que tentam inferir a relação entre a forma clínica da esquistossomose e suas manifestações neurológicas. Objetivo: Avaliar o fluxo sanguíneo do sistema porta em pacientes com mielite esquistossomótica, contribuindo no entendimento dos mecanismos que favorecem a chegada dos parasitas e/ou seus derivados ao sistema nervoso. Método: Foram avaliados 43 indivíduos pela Dopplerfluxometria portal. O grupo I constou de 13 pacientes com esquistossomose medular, sendo o grupo II composto por 10 portadores de formas intestinal e hepatointestinal (sem doença neurológica, e o grupo III, por 20 controles sadios. Resultados: A velocidade venosa portal e o fluxo venoso portal foram significativamente elevados (p-valor<0,001 no grupo I. Conclusão: A Dopplerfluxometria demonstrou a presença de estados hiperdinâmicos do leito vascular porta em pacientes com mielite pelo Schistosoma mansoni.

  4. Humanoid by ROBO-BLOCK

    Science.gov (United States)

    Niimi, Hirofumi; Koike, Minoru; Takeuchi, Seiichi; Douhara, Noriyoshi

    2007-12-01

    Humanoid by ROBO-BLOCK (robot block system) and the rational formula of robots were proposed. ROBO-BLOCK is composed of servo motors, the parts for servomotor rotor, the brackets for servomotor fixation, the board parts and the controllers. A robot can be assembled easily by ROBO-BLOCK. Meanwhile, it is convenient when the structure of the robot can be described easily as a character. The whole structure of the robot is expressed as rational formula of the robot to show molecule structure in chemistry. ROBO-BLOCK can be useful for not only the research but also the education. Creative student experiment was done in the college of industrial technology.

  5. Dicty_cDB: SHB650 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 1 CV682907 |CV682907.1 service43_H06_sjs2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 9e-04... 1 CV682854 |CV682854.1 service30_G09_sjs2-14-10C09-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 9e-... CV688215 |CV688215.1 sjs2-16-2_G01_sjs2-16-2G01-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 9e-04

  6. Dicty_cDB: SHE841 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available ice43_H06_sjs2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 ser...V688215 |CV688215.1 sjs2-16-2_G01_sjs2-16-2G01-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 serv...vice30_G09_sjs2-14-10C09-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001

  7. Dicty_cDB: SHG508 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 1 1 CV682854 |CV682854.1 service30_G09_sjs2-14-10C09-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.... 1 CV682907 |CV682907.1 service43_H06_sjs2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.00...1 CV688215 |CV688215.1 sjs2-16-2_G01_sjs2-16-2G01-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001

  8. Spintronics: Conceptual Building Blocks

    Science.gov (United States)

    Ansermet, J.-Ph.

    The purpose of this introduction to spintronics is to provide some elementary description of its conceptual building blocks. Thus, it is intended for a newcomer to the field. After recalling rudimentary descriptions of spin precession and spin relaxation, spin-dependent transport is treated within the Boltzmann formalism. This suffices to introduce key notions such as the spin asymmetry of the conductivities in the two-current model, the spin diffusion length, and spin accumulation. Two basic mechanisms of spin relaxation are then presented, one arising from spin-orbit scattering and the other from electron-magnon collisions. Finally, the action of a spin-polarized current on magnetization is presented in a thermodynamics framework. This introduces the notion of spin torque and the characteristic length scale over which the transverse spin polarization of conduction electron decays as it is injected into a magnet.

  9. Photovoltaic building blocks

    DEFF Research Database (Denmark)

    Hanberg, Peter Jesper; Jørgensen, Anders Michael

    2014-01-01

    it directcompetitive with fossil energy sources a further reduction is needed. By increasing the efficiency of the solar cells one gain an advantage through the whole chain of cost. So that per produced Watt of power less material is spent, installation costs are lower, less area is used etc. With an average...... efficiency of about 15% for commercial Silicon solar cells there is still much to gain. DTU Danchip provides research facilities, equipment and expertise for the building blocks that comprises fabricating the efficient solar cell. In order to get more of the sun light into the device we provide thin film......Photovoltaics (PV), better known as solar cells, are now a common day sight on many rooftops in Denmark.The installed capacity of PV systems worldwide is growing exponentially1 and is the third most importantrenewable energy source today. The cost of PV is decreasing fast with ~10%/year but to make...

  10. Celiac ganglia block

    Energy Technology Data Exchange (ETDEWEB)

    Akinci, Devrim [Department of Radiology, Hacettepe University School of Medicine, Sihhiye, 06100 Ankara (Turkey); Akhan, Okan [Department of Radiology, Hacettepe University School of Medicine, Sihhiye, 06100 Ankara (Turkey)]. E-mail: oakhan@hacettepe.edu.tr

    2005-09-01

    Pain occurs frequently in patients with advanced cancers. Tumors originating from upper abdominal viscera such as pancreas, stomach, duodenum, proximal small bowel, liver and biliary tract and from compressing enlarged lymph nodes can cause severe abdominal pain, which do not respond satisfactorily to medical treatment or radiotherapy. Percutaneous celiac ganglia block (CGB) can be performed with high success and low complication rates under imaging guidance to obtain pain relief in patients with upper abdominal malignancies. A significant relationship between pain relief and degree of tumoral celiac ganglia invasion according to CT features was described in the literature. Performing the procedure in the early grades of celiac ganglia invasion on CT can increase the effectiveness of the CGB, which is contrary to World Health Organization criteria stating that CGB must be performed in patients with advanced stage cancer. CGB may also be effectively performed in patients with chronic pancreatitis for pain palliation.

  11. Atomic Basic Blocks

    Science.gov (United States)

    Scheler, Fabian; Mitzlaff, Martin; Schröder-Preikschat, Wolfgang

    Die Entscheidung, einen zeit- bzw. ereignisgesteuerten Ansatz für ein Echtzeitsystem zu verwenden, ist schwierig und sehr weitreichend. Weitreichend vor allem deshalb, weil diese beiden Ansätze mit äußerst unterschiedlichen Kontrollflussabstraktionen verknüpft sind, die eine spätere Migration zum anderen Paradigma sehr schwer oder gar unmöglich machen. Wir schlagen daher die Verwendung einer Zwischendarstellung vor, die unabhängig von der jeweils verwendeten Kontrollflussabstraktion ist. Für diesen Zweck verwenden wir auf Basisblöcken basierende Atomic Basic Blocks (ABB) und bauen darauf ein Werkzeug, den Real-Time Systems Compiler (RTSC) auf, der die Migration zwischen zeit- und ereignisgesteuerten Systemen unterstützt.

  12. Establishment of AFLP Reaction System for Cypripedium japonicum Thunb.%扇脉杓兰AFLP反应体系的建立

    Institute of Scientific and Technical Information of China (English)

    李全健; 王彩霞; 田敏; 李翠新

    2012-01-01

    通过SDS法、高盐低pH法、常规CTAB法、高盐CTAB法和改良CTAB法提取扇脉杓兰(Cypripedium japonicum Thunb.)幼叶基因组DNA,并对其AFLP反应体系进行了优化.结果表明:改良CTAB法所提取的DNA电泳条带清晰无污染,A260和A280比值在1.8~2.0,用限制性内切酶酶切后条带均一,酶切时间以4h为宜.最终确定50 μL PCR反应体系的最佳条件为:预扩增产物稀释20倍4μL,dNTP(2.5 mmol/L)3 μL,Mg2+(25 mmol/L)4 μL,Mse I(10 μmol/L)和EcoR I(10 μmol/L)各1.5 μL,Taq DNA聚合酶(5 U/μL)0.3 μL.%Genomic DNA was extracted from young leaves of Cypripedium japonicum Thunb. By SDS method, high salt and low pH method, routine CTAB method, CTAB with high salt method and modified CTAB method, respectively, and an optimized AFLP reaction system was established for Cypripedium japonicum Thunb.. The results showed that DNA electrophoresis strips, which were extracted by the modified CTAB method, were clear and without contamination, and the proportion of A260 to A280 was from 1.8 to 2.0. After double digestion by restriction enzyme, the strips were uniform. 4 h is appropriate for enzyme restriction. The optimum PCR reaction system (50 μL) consisted of 4 μL 20 fold dilution of the pre-amplification product, 3 μL dNTPs (2.5 mmol/L), 4 (iL Mg2+ (25 mmol/L), 1.5 μL Mse I (10 μmol/L), 1.5 μL EcoR I (10 μmol/L) and 0.3 μL Taq DNA polymerase (5 U/μL)

  13. Additional Evaluation of the Point-of-Contact Circulating Cathodic Antigen Assay for Schistosoma mansoni Infection.

    Science.gov (United States)

    Mwinzi, Pauline N M; Kittur, Nupur; Ochola, Elizabeth; Cooper, Philip J; Campbell, Carl H; King, Charles H; Colley, Daniel G

    2015-01-01

    Studies of the urine-based point-of-contact cathodic circulating antigen test (POC-CCA) in Schistosoma mansoni-endemic settings in Africa indicate it has good sensitivity in detecting infections, but in areas of low prevalence, the POC-CCA can be positive for persons who are egg-negative by Kato-Katz stool assays. We examined the POC-CCA assay for: (a) batch-to-batch stability; (b) intra-reader and inter-reader variability; (c) day-to-day variability compared to Kato-Katz stool assays, and (d) to see if praziquantel (PZQ) treatment converted Kato-Katz-negative/POC-CCA positive individuals to POC-CCA negativity. We found essentially no batch-to-batch variation, negligible intra-reader variability (2%), and substantial agreement for inter-reader reliability. Some day-to-day variation was observed over 5 days of urine collection, but less than the variation in Kato-Katz stool assays over 3 days. To evaluate the effect of treatment on Kato-Katz(-)/POC-CCA(+) children, 149 children in an area of 10-15% prevalence who were Kato-Katz(-) based on 3 stool samples but POC-CCA(+) were enrolled. Seven days after treatment (PZQ 40 mg/kg) samples were again collected and tested. Almost half (47%) POC-CCA positive children turned negative. Those still POC-CCA positive received a second treatment, and 34% of them turned POC-CCA negative upon this second treatment. Most who remained POC-CCA positive shifted each time to a "lesser" POC-CCA "level of positivity." The data suggest that most Kato-Katz-negative/POC-CCA positive individuals harbor low-intensity infections, and each treatment kills all or some of their adult worms. The data also suggest that when evaluated by a more sensitive assay, the effective cure rates for PZQ are significantly less than those inferred from fecal testing. These findings have public health significance for the mapping and monitoring of Schistosoma infections and in planning the transition from schistosomiasis morbidity control to elimination of

  14. Emprego da azida sódica, como conservador de fezes, para a pesquisa de ovos de Schistosoma mansoni pelo método de Kato-Katz

    Directory of Open Access Journals (Sweden)

    Elenice Messias do Nascimento Gonçalves

    1988-06-01

    Full Text Available O método de Kato-Katz é muito utilizado para pesquisa de ovos de helmintos nasfezes e, em determinadas ocasiões, como por exemplo no trabalho de campo, afigura-se conveniente preservar o material a examinar, com o intuito de facilitar o transporte e operacionalidade. Na tentativa de poder usar conservador sôliào, capaz de, em relação a ovos de Schistosoma mansoni, manter a morfologia, impedir a evolução e não interferir no processo de clarificação pela glicerina, os autores utilizaram a azida sádica (NaN3, que foi misturada, na quantidade de 2-3mg em aproximadamente 2 g de fezes de pacientes eliminando número conhecido de ovos, quantificados pelo processo de Kato-Katz. As fezes com preservador ficaram mantidas em temperatura ambiente e foram feitas contagens, pela mesma técnica, após uma, duas, quatro, oito e doze semanas. As observações, feitas em 53 amostras, demonstraram que em 51 o número de ovos permaneceu, aproximadamente, idêntico e com estruturas conservadas, de molde a permitir o dignóstico. Em dois casos, nas oitava e décima-segunda semanas, as fezes estavam desidratadas, ressecadas e impróprias para a contagem. A azida sódica, portanto, mostrou-se adequada para a conservação de fezes a serem submetidas ao método de Kato-Katz.The Kato-Katz technique has been widely employed in the investigation of helminthic eggs in faeces. Sometimes, however, stool specimens must be preserved, specially in field surveys, in order to allow transport and improve feasibility. In an attempt to use a solid preservative, capable of keeping the morphology of Schistosoma mansoni eggs intact, blocking parasite development, without interfering with the usual glycerin clarification method, the authors have added 2-3 mg sodium azide (NaN3 to stools. Each specimen contained about 2 g stools and was obtained from patients, who eliminated a known egg burden, determined by the Kato-Katz' technique. Stools with sodium azide were kept at room

  15. Large Block Test Final Report

    Energy Technology Data Exchange (ETDEWEB)

    Lin, W

    2001-12-01

    This report documents the Large-Block Test (LBT) conducted at Fran Ridge near Yucca Mountain, Nevada. The LBT was a thermal test conducted on an exposed block of middle non-lithophysal Topopah Spring tuff (Tptpmn) and was designed to assist in understanding the thermal-hydrological-mechanical-chemical (THMC) processes associated with heating and then cooling a partially saturated fractured rock mass. The LBT was unique in that it was a large (3 x 3 x 4.5 m) block with top and sides exposed. Because the block was exposed at the surface, boundary conditions on five of the six sides of the block were relatively well known and controlled, making this test both easier to model and easier to monitor. This report presents a detailed description of the test as well as analyses of the data and conclusions drawn from the test. The rock block that was tested during the LBT was exposed by excavation and removal of the surrounding rock. The block was characterized and instrumented, and the sides were sealed and insulated to inhibit moisture and heat loss. Temperature on the top of the block was also controlled. The block was heated for 13 months, during which time temperature, moisture distribution, and deformation were monitored. After the test was completed and the block cooled down, a series of boreholes were drilled, and one of the heater holes was over-cored to collect samples for post-test characterization of mineralogy and mechanical properties. Section 2 provides background on the test. Section 3 lists the test objectives and describes the block site, the site configuration, and measurements made during the test. Section 3 also presents a chronology of events associated with the LBT, characterization of the block, and the pre-heat analyses of the test. Section 4 describes the fracture network contained in the block. Section 5 describes the heating/cooling system used to control the temperature in the block and presents the thermal history of the block during the test

  16. Dimensional Reduction for Conformal Blocks

    CERN Document Server

    Hogervorst, Matthijs

    2016-01-01

    We consider the dimensional reduction of a CFT, breaking multiplets of the d-dimensional conformal group SO(d+1,1) up into multiplets of SO(d,1). This leads to an expansion of d-dimensional conformal blocks in terms of blocks in d-1 dimensions. In particular, we obtain a formula for 3d conformal blocks as an infinite sum over 2F1 hypergeometric functions with closed-form coefficients.

  17. Effect of maternal Schistosoma mansoni infection and praziquantel treatment during pregnancy on Schistosoma mansoni infection and immune responsiveness among offspring at age five years.

    Directory of Open Access Journals (Sweden)

    Robert Tweyongyere

    Full Text Available Offspring of Schistosoma mansoni-infected women in schistosomiasis-endemic areas may be sensitised in-utero. This may influence their immune responsiveness to schistosome infection and schistosomiasis-associated morbidity. Effects of praziquantel treatment of S. mansoni during pregnancy on risk of S. mansoni infection among offspring, and on their immune responsiveness when they become exposed to S. mansoni, are unknown. Here we examined effects of praziquantel treatment of S. mansoni during pregnancy on prevalence of S. mansoni and immune responsiveness among offspring at age five years.In a trial in Uganda (ISRCTN32849447, http://www.controlled-trials.com/ISRCTN32849447/elliott, offspring of women treated with praziquantel or placebo during pregnancy were examined for S. mansoni infection and for cytokine and antibody responses to SWA and SEA, as well as for T cell expression of FoxP3, at age five years.Of the 1343 children examined, 32 (2.4% had S. mansoni infection at age five years based on a single stool sample. Infection prevalence did not differ between children of treated or untreated mothers. Cytokine (IFNγ, IL-5, IL-10 and IL-13 and antibody (IgG1, Ig4 and IgE responses to SWA and SEA, and FoxP3 expression, were higher among infected than uninfected children. Praziquantel treatment of S. mansoni during pregnancy had no effect on immune responses, with the exception of IL-10 responses to SWA, which was higher in offspring of women that received praziquantel during pregnancy than those who did not.We found no evidence that maternal S. mansoni infection and its treatment during pregnancy influence prevalence and intensity of S. mansoni infection or effector immune response to S. mansoni infection among offspring at age five years, but the observed effects on IL-10 responses to SWA suggest that maternal S. mansoni and its treatment during pregnancy may affect immunoregulatory responsiveness in childhood schistosomiasis. This might have

  18. Effect of Verticillium lecanii on biological characteristics and life table of Serangium japonicum (Coleoptera: Coccinellidae), a predator of whiteflies under laboratory conditions

    Institute of Scientific and Technical Information of China (English)

    Lazreg Fatiha; Zhen Huang; Shun-Xiang Ren; Shaukat Ali

    2008-01-01

    Effects of entomopathogenic fungus Verticillium lecanii on biological characteristics and life table of Serangiumjaponicum, a predator of whiteflies against five different conidial concentrations (1×104, 1×105, 1×106, 1×107, and 1×108 conidia/mL) were studied under laboratory conditions. The developmental periods for all immature stages (from eggs, 1st, 2nd, 3rd, 4th instar nymph and pupae up to emergence) among the treatments were significantly different when compared to that of control, and the longest development period was observed as treated with 1×108spore/mL. However, no significant difference on the percent survival of all immature stages was observed among the treatments and control. Also, there were no significantly different effects of V. lecanii on mean generation time, intrinsic rate, the finite rate of increase and longevity of S. japonicum among the treatments and control.

  19. Common blocks for ASQS(12

    Directory of Open Access Journals (Sweden)

    Lorenzo Milazzo

    1997-05-01

    Full Text Available An ASQS(v is a particular Steiner system featuring a set of v vertices and two separate families of blocks, B and G, whose elements have a respective cardinality of 4 and 6. It has the property that any three vertices of X belong either to a B-block or to a G-block. The parameter cb is the number of common blocks in two separate ASQSs, both defined on the same set of vertices X . In this paper it is shown that cb ≤ 29 for any pair of ASQSs(12.

  20. Eikonalization of Conformal Blocks

    CERN Document Server

    Fitzpatrick, A Liam; Walters, Matthew T; Wang, Junpu

    2015-01-01

    Classical field configurations such as the Coulomb potential and Schwarzschild solution are built from the $t$-channel exchange of many light degrees of freedom. We study the CFT analog of this phenomenon, which we term the `eikonalization' of conformal blocks. We show that when an operator $T$ appears in the OPE $\\mathcal{O}(x) \\mathcal{O}(0)$, then the large spin $\\ell$ Fock space states $[TT \\cdots T]_{\\ell}$ also appear in this OPE with a computable coefficient. The sum over the exchange of these Fock space states in an $\\langle \\mathcal{O} \\mathcal{O} \\mathcal{O} \\mathcal{O} \\rangle$ correlator build the classical `$T$ field' in the dual AdS description. In some limits the sum of all Fock space exchanges can be represented as the exponential of a single $T$ exchange in the 4-pt correlator of $\\mathcal{O}$. Our results should be useful for systematizing $1/\\ell$ perturbation theory in general CFTs and simplifying the computation of large spin OPE coefficients. As examples we obtain the leading $\\log \\ell$...

  1. Adductor Canal Block versus Femoral Nerve Block and Quadriceps Strength

    DEFF Research Database (Denmark)

    Jæger, Pia Therese; Nielsen, Zbigniew Jerzy Koscielniak; Henningsen, Lene Marianne;

    2013-01-01

    : The authors hypothesized that the adductor canal block (ACB), a predominant sensory blockade, reduces quadriceps strength compared with placebo (primary endpoint, area under the curve, 0.5-6 h), but less than the femoral nerve block (FNB; secondary endpoint). Other secondary endpoints were...

  2. The Bradyrhizobium japonicum fegA gene encodes an iron-regulated outer membrane protein with similarity to hydroxamate-type siderophore receptors.

    Science.gov (United States)

    LeVier, K; Guerinot, M L

    1996-12-01

    Iron is important in the symbiosis between soybean and its nitrogen-fixing endosymbiont Bradyrhizobium japonicum, yet little is known about rhizobial iron acquisition strategies. Analysis of outer membrane proteins (OMPs) from B. japonicum 61A152 identified three iron-regulated OMPs in the size range of several known receptors for Fe(III)-scavenging siderophores. One of the iron-regulated proteins, FegA, was purified and microsequenced, and a reverse genetics approach was used to clone a fegA-containing DNA fragment. Sequencing of this fragment revealed a single open reading frame of 750 amino acids. A putative N-terminal signal sequence of 14 amino acids which would result in a mature protein of 736 amino acids with a molecular mass of 80,851 Da was predicted. FegA shares significant amino acid similarity with several Fe(III)-siderophore receptors from gram-negative bacteria and has greater than 50% amino acid similarity and 33% amino acid identity with two [corrected] bacterial receptors for hydroxamate-type Fe(III)-siderophores. A dendrogram describing total inferred sequence similarity among 36 TonB-dependent OMPs was constructed; FegA grouped with Fe(III)-hydroxamate receptors. The transcriptional start site of fegA was mapped by primer extension analysis, and a putative Fur-binding site was found in the promoter. Primer extension and RNA slot blot analysis demonstrated that fegA was expressed only in cells grown under iron-limiting conditions. This is the first report of the cloning of a gene encoding a putative Fe(III)-siderophore receptor from nitrogen-fixing rhizobia.

  3. Parasitological characteristics of Schistosoma mansoni infection in swiss mice with underlying malnutrition

    Directory of Open Access Journals (Sweden)

    Simões Carla

    2002-01-01

    Full Text Available The effects of a protein-restricted diet (8% protein, 81% carbohydrate and 11% lipids on Schistosoma mansoni infectivity, fecal egg excretion and intestinal egg distribution in Swiss (SW mice were studied. Pregnant mice received a deficient diet from the middle of gestation until delivery. Seven-days-old mice were exposed to 50 cercariae (BH strain, Brazil. Offspring mice had a free access to the deficient diet since lactation until adulthood. The controls were fed with a commercial mice diet. A parasitological examination was performed between six and eight weeks post-infection while both groups were necropsied one week later. Mice on the experimental diet showed a significant loss in body weight. There was no significant difference (p > 0.05 in pre-patent period, kinetics of egg excretion and worm recovery from mice on either diet. Significant differences (p < 0.05 were found concerning to the percentage of deposited eggs in the distal segment of the small intestine from hosts on the experimental diet.Our data suggest that experimental malnutrition induced for a long term has no detrimental effect on the acute schistosomiais infection in SW mice.

  4. Vaccination with antioxidant enzymes confers protective immunity against challenge infection with Schistosoma mansoni

    Directory of Open Access Journals (Sweden)

    Philip T LoVerde

    2004-08-01

    Full Text Available Schistosoma mansoni, an intravascular parasite, lives in a hostile environment in close contact with host humoral and cellular cytotoxic factors. To establish itself in the host, the parasite has evolved a number of immune evasion mechanisms, such as antioxidant enzymes. Our laboratory has demonstrated that the expression of antioxidant enzymes is developmentally regulated, with the highest levels present in the adult worm, the stage least susceptible to immune elimination, and the lowest levels in the larval stages, the most susceptible to immune elimination. Vaccination of mice with naked DNA constructs containing Cu/Zn cytosolic superoxide dismutase (CT-SOD, signal-peptide containing SOD or glutathione peroxidase (GPX showed significant levels of protection compared to a control group. We have further shown that vaccination with SmCT-SOD but not SmGPX results in elimination of adult worms. Anti-oxidant enzyme vaccine candidates offer an advance over existing vaccine strategies that all seem to target the larval developmental stages in that they target adult worms and thus may have therapeutic as well as prophylactic value. To eliminate the potential for cross-reactivity of SmCT-SOD with human superoxide dismutase, we identified parasite-specific epitope-containing peptides. Our results serve as a basis for developing a subunit vaccine against schistosomiasis.

  5. Ultrasound monitoring of structural urinary tract disease in Schistosoma haematobium infection

    Directory of Open Access Journals (Sweden)

    King Charles H

    2002-01-01

    Full Text Available A major advance in our understanding of the natural history of Schistosoma haematobium-related morbidity has come through the introduction of the portable ultrasound machines for non-invasive examination of the kidneys and bladder. With the use of generators or battery packs to supply power in non-clinical field settings, and with the use of instant photography or miniaturized thermal printers to record permanent images, it is possible to examine scores of individuals in endemic communities every day. Broad-based ultrasound screening has allowed better definition of age-specific disease risks in urinary schistosomiasis. Results indicate that urinary tract abnormalities are common (18% overall prevalence in S. haematobium transmission areas, with a 2-4% risk of either severe bladder abnormality or advanced ureteral obstruction. In longitudinal surveys, ultrasound studies have shown that praziquantel and metrifonate therapy are rapidly effective in reversing urinary tract abnormalities among children. The benefits of treating adults are less well known, but research in progress should help to define this issue. Similarly, the prognosis of specific ultrasound findings needs to be clarified, and the ease of sonographic examination will make such long-term follow-up studies feasible. In summary, the painless, quick, and reproducible ultrasound examination has become an essential tool in the study of urinary schistosomiasis.

  6. Detection of Urinary Tract Pathology in Some Schistosoma haematobium Infected Nigerian Adults.

    Science.gov (United States)

    Onile, O S; Awobode, H O; Oladele, V S; Agunloye, A M; Anumudu, C I

    2016-01-01

    Screening for Schistosoma haematobium infection and its possible morbidity was carried out in 257 adult participants in Eggua community, Ogun State, Nigeria. Parasitological assessment for the presence of ova of S. haematobium in urine and abdominopelvic ultrasonographic examination for bladder and secondary kidney pathology were carried out. S. haematobium prevalence of 25.68% (66/257) was recorded among the participants. There was a significantly higher prevalence of 69.2% of urinary schistosomiasis in the females than the prevalence of 31.8% in males (P = 0.902). The intensity of infections was mostly light (55) (21.8%) compared to heavy (10) (3.9%) with the mean intensity of 16.7 eggs/10 mL urine. Structural bladder pathology prevalence among participants was 33.9%. The bladder and kidney pathologies observed by ultrasound in subjects with S. haematobium infections included abnormal bladder wall thickness (59%), abnormal bladder shape (15.2%), bladder wall irregularities (15.2%), bladder masses (1.5%), bladder calcification (1.5%), and hydronephrosis (3%). Infection with S. haematobium was associated with bladder pathology. Higher frequencies of bladder abnormalities were observed more in the participants with light intensity of S. haematobium infection than in those with heavy infection. More bladder pathology was also seen in women than in men, although this was not statistically significant. In conclusion, there is evidence that the development of bladder pathology may be associated with S. haematobium infection.

  7. Circulating anodic antigen for detection of Schistosoma mansoni infection in Egyptian patients.

    Science.gov (United States)

    El-Morshedy, H; Kinosien, B; Barakat, R; Omer, E; Khamis, N; Deelder, A M; Phillips, M

    1996-02-01

    We evaluated the ability of circulating anodic antigen (CAA) to identify infection with Schistosoma mansoni in a prospective cohort study of 257 Egyptian men, 147 with infection diagnosed by repeated Kato thick smears, and 110 without detectable infection. The CAA levels were obtained and the stool examinations were performed two weeks and one, two, four, and six months after praziquantel therapy for infected men. A CAA enzyme-linked immunosorbent assay was repeated twice on subjects who were otherwise negative for schistosomiasis. Circulating anodic antigen was detected in 117 cases, with an overall test sensitivity before treatment of 0.8. Sensitivity was related to the intensity of infection, ranging from 1.00 with > 400 eggs per gram (epg) of feces to 0.60 for those with or = 4. We conclude that CAA has moderate sensitivity and excellent specificity when used to identify infection with schistosomiasis, as well as to monitor the results of therapy after at least one month after treatment.

  8. Increased prevalence of leukocytes and elevated cytokine levels in semen from Schistosoma haematobium-infected individuals.

    Science.gov (United States)

    Leutscher, Peter D C; Pedersen, Mette; Raharisolo, Clairette; Jensen, Jørgen Skov; Hoffmann, Steen; Lisse, Ida; Ostrowski, Sisse R; Reimert, Claus M; Mauclere, Philippe; Ullum, Henrik

    2005-05-15

    In this study, we investigated the seminal inflammatory response to egg infestation of the urogenital organs in 240 semen-donating men aged 15-49 years living in a Schistosoma haematobium-endemic area of Madagascar. In 29 subjects (12%) with excretion of > or =5 ova/ejaculate, leukocytospermia (>10(6) leukocytes/mL) and the presence of seminal lymphocytes and eosinophil leukocytes were each significantly more prevalent than in 74 subjects (31%) who were S. haematobium negative (Pinfection-negative subjects (Pinfection (STI) with Neisseria gonorrhoeae, Chlamydia trachomatis, Mycoplasma genitalium, and/or Trichomonas vaginalis did not act as a confounding factor for the observed associations. At follow-up, 6 months after systematic antischistosomiasis and STI syndrome treatment at baseline, the prevalence of seminal leukocytes decreased significantly among the previously seminal egg-positive subjects. The same tendency was observed for the posttreatment levels of cytokines. Numerous studies have already shown an association between STI-associated genital inflammation and human immunodeficiency virus (HIV) propagation. Therefore, the results of the present study suggest that male urogenital schistosomiasis may constitute a risk factor for HIV transmission, as a result of egg-induced inflammation in the semen-producing pelvic organs.

  9. The extracellular release of Schistosoma mansoni HMGB1 nuclear protein is mediated by acetylation

    Energy Technology Data Exchange (ETDEWEB)

    Coutinho Carneiro, Vitor; Moraes Maciel, Renata de; Caetano de Abreu da Silva, Isabel; Furtado Madeira da Costa, Rodrigo [Instituto de Bioquimica Medica, Programa de Biotecnologia e Biologia Molecular, Universidade Federal do Rio de Janeiro, CCS, Ilha do Fundao, Rio de Janeiro 21941-590 (Brazil); Neto Paiva, Claudia; Torres Bozza, Marcelo [Departamento de Imunologia, Instituto de Microbiologia Professor Paulo de Goes, Universidade Federal do Rio de Janeiro, CCS, Ilha do Fundao, Rio de Janeiro 21941-590 (Brazil); Rosado Fantappie, Marcelo, E-mail: fantappie@bioqmed.ufrj.br [Instituto de Bioquimica Medica, Programa de Biotecnologia e Biologia Molecular, Universidade Federal do Rio de Janeiro, CCS, Ilha do Fundao, Rio de Janeiro 21941-590 (Brazil)

    2009-12-25

    Schistosoma mansoni HMGB1 (SmHMGB1) was revealed to be a substrate for the parasite histone acetyltransferases SmGCN5 and SmCBP1. We found that full-length SmHMGB1, as well as its HMG-box B (but not HMG-box A) were acetylated in vitro by SmGCN5 and SmCBP1. However, SmCBP1 was able to acetylate both substrates more efficiently than SmGCN5. Interestingly, the removal of the C-terminal acidic tail of SmHMGB1 (SmHMGB1{Delta}C) resulted in increased acetylation of the protein. We showed by mammalian cell transfection assays that SmHMGB1 and SmHMGB1{Delta}C were transported from the nucleus to the cytoplasm after sodium butyrate (NaB) treatment. Importantly, after NaB treatment, SmHMGB1 was also present outside the cell. Together, our data suggest that acetylation of SmHMGB1 plays a role in cellular trafficking, culminating with its secretion to the extracellular milieu. The possible role of SmHMGB1 acetylation in the pathogenesis of schistosomiasis is discussed.

  10. Scanning electron microscopic study of the tegumental surface of adult Schistosoma sinensium.

    Science.gov (United States)

    Kruatrachue, M; Upatham, E S; Sahaphong, S; Tongthong, T; Khunborivan, V

    1983-12-01

    The SEM study of tegumental surface of adult Schistosoma sinensium reveals that the male tegument lacks tubercles or bosses; instead it is corrugated with small pits or perforated ridges. On the dorsal surface, spines are present whose number and size progressively increase towards the posterior end of the body. In addition, there are three types of papillae interspersed among the ridges and spines. The first type of papillae has crater-like holes surrounded by a circular doughnut-shaped elevation; some are ciliated and others are non-ciliated. They are generally found on the dorsal and ventral surfaces. The second is sensory papillae which are hemispherical in shape bearing apical cilia. They are found to be concentrated around the oral sucker and on the posterior end of the worm. The third is fungiform papillae without cilia which are found on the posterior end. There are short spines present on the tegument lining the gynecophoral canal of the male worm. The tegument of the female S. sinensium is corrugated with ridges on the ventral surface. Small spines are present on the anterior portion of the dorsal surface. They become larger and increased in number towards the posterior end of the worm. The three types of papillae are present but they are much fewer and less developed than those in the male worm.

  11. Therapy with bone marrow cells reduces liver alterations in mice chronically infected by Schistosoma mansoni

    Institute of Scientific and Technical Information of China (English)

    Sheilla Andrade Oliveira; Bruno Solano Freitas Souza; Cada Adriana Guimar(a)es-Ferreira; Elton Sá Barreto; Siane Campos Souza; Luiz Antonio Rodrigues Freitas; Ricardo Ribeiro-dos-Santos; Milena Botelho Pereira Soares

    2008-01-01

    AIM: To investigate the potential of bone marrow mononuclear cells (BM-MCs) in the regeneration of hepatic lesions induced by Schistosoma mansoni (S.mansoni) chronic infection.METHODS: Female mice chronically infected with S.mansoni were treated with BM-MCs obtained from male green fluorescent protein (GFP) transgenic mice by intravenous or intralobular injections. Control mice received injections of saline in similar conditions. Enzyme-linked immunosorbent assay (ELISA) assay for transforming growth factor-beta (TGF-β), polymerase chain reaction (PCR) for GFP DNA, immunofluorescence and morphometric studies were performed.RESULTS: Transplanted GFP+ cells migrated to granuloma areas and reduced the percentage of liver fibrosis. The presence of donor-derived cells was confirmed by Fluorescence in situ hybridization (FISH) analysis for detection of cells bearing Y chromosome and by PCR analysis for detection of GFP DNA. The levels of TGF-β, a cytokine associated with fibrosis deposition, in liver fragments of mice submitted to therapy were reduced. The number of oval cells in liver sections of S.rnansoni-infected mice increased 3-4 fold after transplantation. A partial recovery in albumin expression, which is decreased upon infection with S.mansoni, was found in livers of infected mice after cellular therapy.CONCLUSION: In conclusion, transplanted BMCs migrate to and reduce the damage of chronic fibrotic liver lesions caused by S.mansoni.

  12. Experimental Evaluation of the Pathogenicity of Different Strains of Schistosoma mansoni

    Directory of Open Access Journals (Sweden)

    Antônio Aurélio Euzébio

    2012-01-01

    Full Text Available The pathogenesis of three different Schistosoma mansoni strains from the Brazilian states of Minas Gerais (BH strain and São Paulo (SJ and SD strains was evaluated in experimentally infected mice. Observations of the most severe clinical cases among local patients treated (SD strain in the city of Campinas (São Paulo, Brazil formed the basis of this study. Mice were used as definitive hosts and were infected with cercariae from Biomphalaria tenagophila (SJ and SD strains and Biomphalaria glabrata (BH strains. The parameters analyzed were as follows: number of S. mansoni eggs in mice feces; number of granulomas per tissue area in liver, spleen, lungs, pancreas, and ascending colon; measurements of hepatic and intestinal granulomas; number of adult worms; and measurements of trematode eggs. The comparison among the three strains indicated that the SD strain, isolated in Campinas, presented a higher worm recovery relative to the number of penetrating cercariae. In addition, when compared to the SJ and BH strains, the SD strain demonstrated similar pathogenicity to the BH strain, with a greater quantity of granulomas in the viscera, as well as larger granulomas and eggs. Furthermore, a greater quantity of trematode eggs was also shed in the feces.

  13. Experimental Evaluation of the Pathogenicity of Different Strains of Schistosoma mansoni.

    Science.gov (United States)

    Euzébio, Antônio Aurélio; Zuim, Nádia Regina Borim; Linhares, Arício Xavier; Magalhães, Luiz Augusto; Zanotti-Magalhães, Eliana Maria

    2012-01-01

    The pathogenesis of three different Schistosoma mansoni strains from the Brazilian states of Minas Gerais (BH strain) and São Paulo (SJ and SD strains) was evaluated in experimentally infected mice. Observations of the most severe clinical cases among local patients treated (SD strain) in the city of Campinas (São Paulo, Brazil) formed the basis of this study. Mice were used as definitive hosts and were infected with cercariae from Biomphalaria tenagophila (SJ and SD strains) and Biomphalaria glabrata (BH strains). The parameters analyzed were as follows: number of S. mansoni eggs in mice feces; number of granulomas per tissue area in liver, spleen, lungs, pancreas, and ascending colon; measurements of hepatic and intestinal granulomas; number of adult worms; and measurements of trematode eggs. The comparison among the three strains indicated that the SD strain, isolated in Campinas, presented a higher worm recovery relative to the number of penetrating cercariae. In addition, when compared to the SJ and BH strains, the SD strain demonstrated similar pathogenicity to the BH strain, with a greater quantity of granulomas in the viscera, as well as larger granulomas and eggs. Furthermore, a greater quantity of trematode eggs was also shed in the feces.

  14. Characterization of export receptor exportins (XPOs) in the parasite Schistosoma mansoni.

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    Abreu, Fabiano C P; Pereira, Roberta V; Oliveira, Victor F; Gomes, Matheus de S; Jannotti-Passos, Liana K; Borges, William C; Guerra-Sá, Renata

    2013-12-01

    Several proteins and different species of RNA that are produced in the nucleus are exported through the nuclear pore complexes, which require a family of conserved nuclear export receptors called exportins (XPOs). It has been reported that the XPOs (XPO1, XPO5, and XPOT) are directly involved in the transport processes of noncoding RNAs from the nucleus to the cytoplasm and/or from cytoplasm to the nucleus. All three genes are present in fungi, plants, and deuterostome metazoans. However, protostome metazoan species lack one of the three genes across evolution. In this report, we have demonstrated that all three XPO proteins are present in the parasite protostome Schistosoma mansoni. As this parasite has a complex life cycle presenting several stages in different hosts and environments, implying a differential gene regulation, we proposed a genomic analysis of XPOs to validate their annotation. The results showed the conservation of exportin family members and gene duplication events in S. mansoni. We performed quantitative RT-PCR, which revealed an upregulation of SmXPO1 in 24 h schistosomula (sixfold when compared with cercariae), and similar transcription levels were observed for SmXPO5 and SmXPOT in all the analyzed stages. These three XPO proteins have been identified for the first time in the protostome clade, which suggests a higher complexity in RNA transport in the parasite S. mansoni. Taken together, these results suggest that RNA transport by exportins might control cellular processes during cercariae, schistosomula, and adult worm development.

  15. Antischistosomal activity of a calcium channel antagonist on schistosomula and adult Schistosoma mansoni worms

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    Vanessa Silva-Moraes

    2013-08-01

    Full Text Available Current schistosomiasis control strategies are largely based on chemotherapeutic agents and a limited number of drugs are available today. Praziquantel (PZQ is the only drug currently used in schistosomiasis control programs. Unfortunately, this drug shows poor efficacy in patients during the earliest infection phases. The effects of PZQ appear to operate on the voltage-operated Ca2+channels, which are located on the external Schistosoma mansoni membrane. Because some Ca2+channels have dihydropyridine drug class (a class that includes nifedipine sensitivity, an in vitro analysis using a calcium channel antagonist (clinically used for cardiovascular hypertension was performed to determine the antischistosomal effects of nifedipine on schistosomula and adult worm cultures. Nifedipine demonstrated antischistosomal activity against schistosomula and significantly reduced viability at all of the concentrations used alone or in combination with PZQ. In contrast, PZQ did not show significant efficacy when used alone. Adult worms were also affected by nifedipine after a 24 h incubation and exhibited impaired motility, several lesions on the tegument and intense contractility. These data support the idea of Ca2+channels subunits as drug targets and favour alternative therapeutic schemes when drug resistance has been reported. In this paper, strong arguments encouraging drug research are presented, with a focus on exploring schistosomal Ca2+channels.

  16. Isoforms of Hsp70-binding human LDL in adult Schistosoma mansoni worms.

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    Pereira, Adriana S A; Cavalcanti, Marília G S; Zingali, Russolina B; Lima-Filho, José L; Chaves, Maria E C

    2015-03-01

    Schistosoma mansoni is one of the most common parasites infecting humans. They are well adapted to the host, and this parasite's longevity is a consequence of effective escape from the host immune system. In the blood circulation, lipoproteins not only help to conceal the worm from attack by host antibodies but also act as a source of lipids for S. mansoni. Previous SEM studies showed that the low-density lipoprotein (LDL) particles present on the surface of adult S. mansoni worms decreased in size when the incubation time increased. In this study, immunocytochemical and proteomic analyses were used to locate and identify S. mansoni binding proteins to human plasma LDL. Ultrathin sections of adult worms were cut transversely from the anterior, medial and posterior regions of the parasite. Immunocytochemical experiments revealed particles of gold in the tegument, muscle region and spine in male worms and around vitelline cells in females. Immunoblotting and 2D-electrophoresis using incubations with human serum, anti-LDL antibodies and anti-chicken IgG peroxidase conjugate were performed to identify LDL-binding proteins in S. mansoni. Analysis of the binding proteins using LC-MS identified two isoforms of the Hsp70 chaperone in S. mansoni. Hsp70 is involved in the interaction with apoB in the cytoplasm and its transport to the endoplasmic reticulum. However, further studies are needed to clarify the functional role of Hsp70 in S. mansoni, mainly related to the interaction with human LDL.

  17. Anti-Inflammatory Properties of Menthol and Menthone in Schistosoma mansoni Infection

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    Zaia, Mauricio G.; Cagnazzo, Túlio di Orlando; Feitosa, Karina A.; Soares, Edson G.; Faccioli, Lúcia H.; Allegretti, Silmara M.; Afonso, Ana; Anibal, Fernanda de Freitas

    2016-01-01

    Schistosomiasis is a parasitic disease caused by several species of trematode worms and it is believed that more than 261 million people are affected worldwide. New drug development has become essential because there is a risk of the parasite becoming resistant to Praziquantel, the only drug available for this infection. This study evaluated parasitological, immunological and histological parameters in mice infected with Schistosoma mansoni and treated with an herbal commercial medicine. This drug consists of menthol (30–55%) and menthone (14–32%). A 60 day treatment regimen with the herbal medicine decreased the number of S. mansoni eggs in the feces, liver, and intestine and reduced the number of hepatic granulomas. We observed a reduction of 84% in blood eosinophilia and a decrease in the IL-4 and IL-10 blood levels after treatment. Therefore, we propose that schistosomiasis treatment with this herbal medicine for 60 days has an immunomodulatory and anti-inflammatory action in this animal model for schistosomiasis thus contributing to the decrease in physio pathological effects caused by S. mansoni infection. PMID:27378927

  18. La queratotaxia cercariana en la diferenciacion de sexos de schistosoma mansoni

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    Luz Arelis Pino

    1988-09-01

    Full Text Available El número de papilas argirófilas superficiales y su modelo de disposición en el tegumento de las cercarias (quetotaxia de Schistosoma mansoni nos permitió diferenciar los sexos a nivel del mencionado estadío larvario, mediante los siguientes critérios: - Mayor homogeneidad en las cercarias machos, en cuanto al número total de papilas ventrales y dorsales a nivel de cuerpo y cola cercarianos (C.V. = 4,1%, que en las cercarias hembras (C. V. = 18,3% (P < 0,001. - Presencia en el 80% de las cercarias machos de cuatro papilas en los cuadrantes "C" ó "D" inferior-izquierdo e inferior-derecho, respectivamente ventrales, mientras que dicho caracter sólo está presente en el 40% de las cercarias hembras (P < 0,001. - Diferencia estadísticamente significativa (P < 0,001 entre el número total promedio de papilas corporales centrales de las cercarias hembras (X = 11,9 ± 0,2 y el de las cercarias machos (X = 11,1 ± 0,3. - Diferencia estadísticamente significativa (P < 0,05 para la mayor distancia promedio entre las papilas AIL y AIIL, de cada cercaria, en relación con el sexo (femenino = 25,5 µm ± 0,33; masculino = 27,3 µm ± 0,26.

  19. Vaccination with Enzymatically Cleaved GPI-Anchored Proteins from Schistosoma mansoni Induces Protection against Challenge Infection

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    Vicente P. Martins

    2012-01-01

    Full Text Available The flatworm Schistosoma mansoni is a blood fluke parasite that causes schistosomiasis, a debilitating disease that occurs throughout the developing world. Current schistosomiasis control strategies are mainly based on chemotherapy, but many researchers believe that the best long-term strategy to control schistosomiasis is through immunization with an antischistosomiasis vaccine combined with drug treatment. In the search for potential vaccine candidates, numerous tegument antigens have been assessed. As the major interface between parasite and mammalian host, the tegument plays crucial roles in the establishment and further course of schistosomiasis. Herein, we evaluated the potential of a GPI fraction, containing representative molecules located on the outer surface of adult worms, as vaccine candidate. Immunization of mice with GPI-anchored proteins induced a mixed Th1/Th2 type of immune response with production of IFN-γ and TNF-α, and low levels of IL-5 into the supernatant of splenocyte cultures. The protection engendered by this vaccination protocol was confirmed by 42% reduction in worm burden, 45% reduction in eggs per gram of hepatic tissue, 29% reduction in the number of granulomas per area, and 53% reduction in the granuloma fibrosis. Taken together, the data herein support the potential of surface-exposed GPI-anchored antigens from the S. mansoni tegument as vaccine candidate.

  20. Mechanisms of evasion of Schistosoma mansoni schistosomula to the lethal activity of complement

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    F. Juarez Ramalho-Pinto

    1992-01-01

    Full Text Available Schistosomula of Schistosoma mansoni became resistant to antibody-dependent complement damage in vitro after pre-incubation with normal human erythrocytes (NHuE whatever the ABO or Rh blood group. Resistant parasites were shown to acquire host decay accelerating factor (DAF , a 70 kDa glycoprotein attached to the membrane of NHue by a GPI anchor. IgG2a mAb anti-human DAF (IA10 immunoprecipitated a 70 kDa molecule from 125I-labeled schistosomula pre-incubated with NHuE and inhibited their resistance to complement-dependent killing in vtro. Incubationof schistosomula with erytrocytes from patients with paroxsimal nocturnal hemoglobinuria (PNHE or SRBC, wich are DAF-deficient, did not protect the parasites from complement lesion. Supernatant of 100,000 x g collected from NHuE incubated for 24 h in defined medium was shown to contain a soluble form of DAF and to protect schistosomula from complement killing. Schistosomula treated with trypsin before incubation with NHuE ghosts did not become resistant to complement damage. On the other hand, pre-treatment with chymotrypsin did not interfere with the acquisition of resistance by the schistosomula. These results indicate that, in vitro, NHuE DAF can be transferred to schistosomula in a soluble form and that the binding of this molecule to the parasite surface is dependent upon trypsin-sensitive chymotrypsin-insensitive polipeptide(s present on the surface of the worm.

  1. Extra-cellular matrix changes in Schistosoma mansoni-infected Biomphalaria glabrata

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    Borges Claudia Maria da Cunha

    2003-01-01

    Full Text Available Reactivity of snails against parasites exhibits a primitive focal reaction, with encapsulation, phagocytosis and destruction of parasite larvae by macrophage-like cells - the hemocytes. This reaction mimics granulomatous inflammation seen in higher animals. However, different from the latter, little is known about the participation of extra-cellular matrix in such snail defense reactions. Normal and Schistosoma mansoni-infected Biomphalaria glabrata of different strains were submitted to cytological, histological, ultrastructural and biochemical methods in order to investigate the behavior of extra-cellular tissues at the site of anti-parasite reactions. In spite of the presence of two cell-types in peripheral hemolymph, only one cell-type was present at the sites of tissue reactions. Although pre-existent collagen and elastic fibers and microfibrils sometimes appeared slightly compressed around focal reactions, no evidences of duplication, synthesis or deposition of connective-tissue extra-cellular components were observed within or around the zones of reactive cell accumulations. Thus, tissue reactions against S. mansoni in the snail B. glabrata appeared exclusively dependent on one specific population of hemocytes.

  2. A bacterial artificial chromosome library for Biomphalaria glabrata, intermediate snail host of Schistosoma mansoni

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    Coen M Adema

    2006-10-01

    Full Text Available To provide a novel resource for analysis of the genome of Biomphalaria glabrata, members of the international Biomphalaria glabrata Genome Initiative (biology.unm.edu/biomphalaria-genome.html, working with the Arizona Genomics Institute (AGI and supported by the National Human Genome Research Institute (NHGRI, produced a high quality bacterial artificial chromosome (BAC library. The BB02 strain B. glabrata, a field isolate (Belo Horizonte, Minas Gerais, Brasil that is susceptible to several strains of Schistosoma mansoni, was selfed for two generations to reduce haplotype diversity in the offspring. High molecular weight DNA was isolated from ovotestes of 40 snails, partially digested with HindIII, and ligated into pAGIBAC1 vector. The resulting B. glabrata BAC library (BG_BBa consists of 61824 clones (136.3 kb average insert size and provides 9.05 × coverage of the 931 Mb genome. Probing with single/low copy number genes from B. glabrata and fingerprinting of selected BAC clones indicated that the BAC library sufficiently represents the gene complement. BAC end sequence data (514 reads, 299860 nt indicated that the genome of B. glabrata contains ~ 63% AT, and disclosed several novel genes, transposable elements, and groups of high frequency sequence elements. This BG_BBa BAC library, available from AGI at cost to the research community, gains in relevance because BB02 strain B. glabrata is targeted whole genome sequencing by NHGRI.

  3. Anthelmintic effects of the essential oil of fennel (Foeniculum vulgare Mill., Apiaceae) against Schistosoma mansoni.

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    Wakabayashi, Kamila A L; de Melo, Nathalya I; Aguiar, Daniela P; de Oliveira, Pollyanna F; Groppo, Milton; da Silva Filho, Ademar A; Rodrigues, Vanderlei; Cunha, Wilson R; Tavares, Denise C; Magalhães, Lizandra G; Crotti, Antônio E M

    2015-07-01

    Foeniculum vulgare Mill. (Apiaceae), known as fennel, is a widespread aromatic herbaceous plant, and its essential oil is used as additive in the food, pharmaceutical, cosmetic, and perfume industries. The in vitro antischistosomal activity and cytotoxic effects against V79 cells of the essential oil of F. vulgare cultivated in southeastern Brazil (FV-EO) was investigated. The FV-EO was obtained by hydrodistillation and characterized by GC-FID and GC/MS analyses. (E)-Anethole (69.8%) and limonene (22.5%) were identified as the major constituents. Its anthelmintic activity against Schistosoma mansoni was evaluated at concentrations of 10, 50, and 100 μg/ml, and it was found to be active against adult S. mansoni worms, although it was less effective than the positive control praziquantel (PZQ) in terms of separation of the coupled pairs, mortality, and decreased motor activity. However, FV-EO elicited an interesting dose-dependent reduction in the number of S. mansoni eggs. On their own, (E)-anethole and the limonene enantiomers were much less effective than FV-EO and PZQ. An XTT-cytotoxicity-based assay evidenced no FV-EO cytotoxicity against V79 cells. In summary, FV-EO displayed moderate in vitro schistosomicidal activity against adult S. mansoni worms, exerted remarkable inhibitory effects on the egg development, and was of low toxicity.

  4. Polymyxin B as inhibitor of LPS contamination of Schistosoma mansoni recombinant proteins in human cytokine analysis

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    Pacífico Lucila G

    2007-01-01

    Full Text Available Abstract Background Recombinant proteins expressed in Escherichia coli vectors are generally contaminated with endotoxin. In this study, we evaluated the ability of Polymyxin B to neutralize the effect of LPS present as contaminant on Schistosoma mansoni recombinant proteins produced in E. coli in inducing TNF-α and IL-10. Peripheral blood mononuclear cells from individuals chronically infected with S. mansoni were stimulated in vitro with recombinant Sm22.6, Sm14 and P24 antigens (10 μg/mL in the presence of Polymyxin B (10 μg/mL. Results The levels of cytokines were measured using ELISA. There was greater than 90 % reduction (p S. mansoni recombinant proteins in the presence of Polymyxin B, a reduction in the levels of TNF-α and IL-10 was also observed. However, the percentage of reduction was lower when compared to the cultures stimulated with LPS, probably because these proteins are able to induce the production of these cytokines by themselves. Conclusion This study showed that Polymyxin B was able to neutralize the effect of endotoxin, as contaminant in S. mansoni recombinant antigens produced in E. coli, in inducing TNF-α and IL-10 production.

  5. Functional visualization of the excretory system of adult Schistosoma mansoni by the fluorescent marker resorufin.

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    Sato, H; Kusel, J R; Thornhill, J

    2002-12-01

    Excretion of metabolic wastes as well as xenobiotics is a major concern of all living organisms, and the Platyhelminthes including Schistosoma mansoni possess the protonephridial excretory system for their survival. Except for some ultra-structural and biochemical information, little is known about the protonephridium of platyhelminths due to a lack of established techniques for exploring the excretory activity. This study describes a new technique to assess the excretory activity of S. mansoni by use of the fluorescent marker resorufin, which is a potential substrate of the drug efflux pump, P-glycoprotein. After simple diffusion into the schistosome body, fluorescent resorufin was concentrated in the excretory tubules by an energy-dependent mechanism and excreted via the nephridiopore. The present technique of labelling functionally the excretory system was applicable to adult worms, but not schistosomula or cercariae. A variety of modulators known to interfere with mammalian P-glycoprotein function perturbed resorufin excretion from male adult schistosomes, including cyclosporin A, Ro11-2933, verapamil, or nifedipine. This technique of labelling the excretory system with fluorescent resorufin has enabled us to study aspects of the physiological function, hitherto unknown, of the protonephridial system of S. mansoni.

  6. Schistosoma mansoni infection modulates the immune response against allergic and auto-immune diseases

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    Maria Ilma Araújo

    2004-08-01

    Full Text Available Chronic Schistosoma mansoni infection leads to a type 2-immune response with increased production of interleukin (IL-10. Evidence indicates chronic exposure to S. mansoni down regulates the type 1 immune response and prevents the onset of Th1-mediated diseases such as multiple sclerosis, diabetes mellitus and Cronh's disease. Furthermore, our own studies have revealed that chronic exposure to S. mansoni also down regulates atopic disease, Th2-mediated diseases. Our studies show an inverse association between the skin prick test reactivity and infection with S. mansoni and show the severity of asthma is reduced in subjects living in an endemic area of S. mansoni. Moreover, we hypothesize the mechanisms involved in the modulation of inflammatory response in atopic individuals, is likely dependent on IL-10 production, an anti-inflammatory cytokine elevated during helminth infections. Patients with asthma and helminth infections produced less IL-5 than patients with asthma without helminth infections, and this down regulation could, in part, be mediated by IL-10. In conclusion, helminthic infections, through induction of regulatory mechanisms, such as IL-10 production, are able to modulate the inflammatory immune response involved in the pathology of auto-immune and allergic disease.

  7. Flavonoids and Sesquiterpene Lactones from Artemisia absinthium and Tanacetum parthenium against Schistosoma mansoni Worms

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    Luísa Maria Silveira de Almeida

    2016-01-01

    Full Text Available Human schistosomiasis, caused by trematode worms of the genus Schistosoma, is one of the most significant neglected tropical diseases, affecting more than 200 million individuals worldwide and praziquantel is the only available drug to treat this disease. Artemisia absinthium L. and Tanacetum parthenium L. are species popularly used as anthelmintics. We investigated the in vitro schistosomicidal activity of crude extracts of A. absinthium (AA and T. parthenium (TP and their isolated compounds. AA and TP, at 200 μg/mL, were active, causing 100% mortality of all adult worms. Chromatographic fractionation of AA leads to isolation of artemetin and hydroxypelenolide, while santin, apigenin, and parthenolide were isolated from TP. Artemetin, hydroxypelenolide, santin, and apigenin, at 100 μM, were inactive against adult worms. Parthenolide (12.5 to 100 μM caused 100% mortality, tegumental alterations, and reduction of motor activity of all adult worms of S. mansoni, without affecting mammalian cells. Confocal laser scanning microscopy showed tegumental morphological alterations and changes on the numbers of tubercles of S. mansoni worms. This report provides the first evidence for the in vitro activity of parthenolide against adult worms of S. mansoni, opening the route to further schistosomicidal studies with this compound.

  8. Predicting the spatial distribution of Biomphalaria straminea, a potential intermediate host for Schistosoma mansoni, in China

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    Mohamed R. Habib

    2016-11-01

    Full Text Available Schistosomiasis is one of the most prevalent parasitic diseases impacting human health in the tropics and sub-tropics. The geographic distribution of Schistosoma mansoni, the most widespread species, includes areas in Africa, the Middle East, South America and the Caribbean. Snails of the genus Biomphalaria act as intermediate host for S. mansoni. Biomphalaria straminea is not indigenous in China but was accidentally introduced to Hong Kong from South America and has spread to other habitats in the southern parts of the country. This species is known for its great dispersal capacity that highlights the importance of the snail as a potential host for S. mansoni in China. In this connection, although no such infection has been recorded in the field so far, the continuous expansion of China’s projects in endemic areas of Africa and import of the infection via returning workers or visitors deserve attention. The purpose of this study was to map and predict the spatial distribution of B. straminea in China. Snail occurrence data were assembled and investigated using MaxEnt software, along with climatic and environmental variables to produce a predictive risk map. Of the environmental variables tested, the precipitation of warmest quarter was the most contribution factor for snail’s spatial distribution. Risk areas were found in southeastern China and it is expected that they will guide policies and control programmes to potential areas area of snail abundance and used for spatial targeting of control interventions for this invasive species.

  9. Polymerase Chain Reaction: A Better Method for Diagnosing Chronic Schistosoma mansoni Infections.

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    Abdel-Hafeez, Ekhlas Hamed; Mohamed, Rabie M; Belal, Usama S; Abdel-Raheem, Ehab M; Naoi, Koji; Norose, Kazumi

    2015-12-01

    For more effective diagnosis of the acute and chronic stages of Schistosoma mansoni infection in humans, the polymerase chain reaction (PCR) technique was compared with the Kato-Katz method. A total of 150 stool samples were collected from inpatient and outpatient clinics at the Department of Tropical Medicine, Minia University Hospital, Egypt. Three groups of patients, 50 with acute intestinal schistosomiasis, 70 with chronic intestinal schistosomiasis and 30 normal healthy controls were studied. Stool samples were analyzed by PCR and the Kato-Katz method. The mean number of eggs per gram of feces was 4.6 when estimated by the Kato-Katz method in positive stool samples from acute schistosomiasis cases but only 1.7 in chronic cases. In acute intestinal schistosomiasis, 15 and 45 out of 50 cases were positive by Kato-Katz and PCR, respectively. In the chronic intestinal schistosomiasis cases, 6 and 68 out of 70 cases were positive by the Kato-Katz and PCR methods, respectively. We conclude that PCR appears to be an effective diagnostic technique for S. mansoni infection, especially where a low worm burden exists, such as in chronic cases.

  10. Schistosoma mansoni: quantification of skin penetration and early migration by differential external radioassay and autoradiography

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    Georgi, J.R. (New York State College of Veterinary Medicine, Ithaca (USA))

    1982-04-01

    Eleven gamma-emitting radionuclides (/sup 49/Sc, /sup 54/Mn, /sup 59/Fe, /sup 60/Co, /sup 65/Zn, /sup 75/Se (as selenomethionine, selenocystine, selenite and selenate), /sup 109/Cd, /sup 125/Sb, /sup 133/Ba, /sup 137/Cs and /sup 203/Hg) were screened as labelling agents for Schistosoma mansoni cercariae by incubation of infected Biomphalaria glabrata snails in radioactive solution. Only (/sup 75/Se)methionine yielded satisfactorily labelled cercariae. Differential external radioassay, a new technique employing partial body shielding within a total body counter, permitted separate estimation of tail and body radioactivity of conscious mice previously exposed by tail immersion to /sup 75/Se-labelled cercariae, with measurements repeated as often as desired. Approximately 39% of the /sup 75/Se present in emergent cercariae was retained by schistosomula transformed in vitro but this was subject to considerable variation, especially in schistosomula transformed in vivo. Secreted or catabolized label from penetrant cercariae and schistosomula was rapidly removed from the skin by the bloodstream. Numbers of schistosomula in tail skin were directly proportional to the number of reduced silver foci counted on tail autoradiograms; only a very small fraction of tail radioactivity represented unbound ('spurious') label. Migration of schistosomula away from skin was 50% complete at 3.8-4.3 days, as determined by probit analysis of autoradiographic data.

  11. Comparison between morphological and staining characteristics of live and dead eggs of Schistosoma mansoni

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    AK Sarvel

    2006-10-01

    Full Text Available Schistosoma mansoni eggs are classified, according to morphological characteristics, as follows: viable mature and immature eggs; dead mature and immature eggs, shells and granulomas. The scope of this study was to compare the staining characteristics of different morphological types of eggs in the presence of fluorescent labels and vital dyes, aiming at differentiating live and dead eggs. The eggs were obtained from the intestines of infected mice, and put into saline 0.85%. The fluorescent labels were Hoechst 33258 and Acridine Orange + Ethidium Bromide and vital dyes (Trypan Blue 0.4% and Neutral Red 1%. When labelled with the probe Hoechst 33258, some immature eggs, morphologically considered viable, presented fluorescence (a staining characteristic detected only in dead eggs; mature eggs did not present fluorescence, and the other types of dead eggs, morphologically defined, showed fluorescence. As far as Acridine Orange + Ethidium Bromide are concerned, either the eggs considered to be live, or the dead ones, presented staining with green color, and only the hatched and motionless miracidium was stained with an orange color. Trypan Blue was not able to stain the eggs, considered to be dead but only dead miracidia which had emerged out of the shell. Neutral Red stained both live and dead eggs. Only the fluorescent Hoechst 33258 can be considered a useful tool for differentiation between dead and live eggs.

  12. Towards an understanding of the function of the phytochelatin synthase of Schistosoma mansoni.

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    Rigouin, Coraline; Nylin, Elyse; Cogswell, Alexis A; Schaumlöffel, Dirk; Dobritzsch, Dirk; Williams, David L

    2013-01-01

    Phytochelatin synthase (PCS) is a protease-like enzyme that catalyzes the production of metal chelating peptides, the phytochelatins, from glutathione (GSH). In plants, algae, and fungi phytochelatin production is important for metal tolerance and detoxification. PCS proteins also function in xenobiotic metabolism by processing GSH S-conjugates. The aim of the present study is to elucidate the role of PCS in the parasitic worm Schistosoma mansoni. Recombinant S. mansoni PCS proteins expressed in bacteria could both synthesize phytochelatins and hydrolyze various GSH S-conjugates. We found that both the N-truncated protein and the N- and C-terminal truncated form of the enzyme (corresponding to only the catalytic domain) work through a thiol-dependant and, notably, metal-independent mechanism for both transpeptidase (phytochelatin synthesis) and peptidase (hydrolysis of GSH S-conjugates) activities. PCS transcript abundance was increased by metals and xenobiotics in cultured adult worms. In addition, these treatments were found to increase transcript abundance of other enzymes involved in GSH metabolism. Highest levels of PCS transcripts were identified in the esophageal gland of adult worms. Taken together, these results suggest that S. mansoni PCS participates in both metal homoeostasis and xenobiotic metabolism rather than metal detoxification as previously suggested and that the enzyme may be part of a global stress response in the worm. Because humans do not have PCS, this enzyme is of particular interest as a drug target for schistosomiasis.

  13. Exposure to Hycanthone alters chromatin structure around specific gene functions and specific repeats in Schistosoma mansoni

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    David eRoquis

    2014-07-01

    Full Text Available Schistosoma mansoni is a parasitic plathyhelminth responsible for intestinal schistosomiasis (or bilharziasis, a disease affecting 67 million people worldwide and causing an important economic burden. The schistosomicides hycanthone, and its later proxy oxamniquine, were widely used for treatments in endemic areas during the 20th century. Recently, the mechanism of action, as well as the genetic origin of a stably and Mendelian inherited resistance for both drugs was elucidated in two strains. However, several observations suggested early on that alternative mechanisms might exist, by which resistance could be induced for these two drugs in sensitive lines of schistosomes. This induced resistance appeared rapidly, within the first generation, but was metastable (not stably inherited. Epigenetic inheritance could explain such a phenomenon and we therefore re-analyzed the historical data with our current knowledge of epigenetics. In addition, we performed new experiments such as ChIP-seq on hycanthone treated worms. We found distinct chromatin structure changes between sensitive worms and induced resistant worms from the same strain. No specific pathway was discovered, but genes in which chromatin structure modification were observed are mostly associated with transport and catabolism, which makes sense in the context of the elimination of the drug. Specific differences were observed in the repetitive compartment of the genome. We finally describe what types of experiments are needed to understand the complexity of heritability that can be based on genetic and/or epigenetic mechanisms for drug resistance in schistosomes.

  14. The genomic proliferation of transposable elements in colonizing populations: Schistosoma mansoni in the new world.

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    Wijayawardena, Bhagya K; DeWoody, J Andrew; Minchella, Dennis J

    2015-06-01

    Transposable elements (TEs) are mobile genes with an inherent ability to move within and among genomes. Theory predicts that TEs proliferate extensively during physiological stress due to the breakdown of TE repression systems. We tested this hypothesis in Schistosoma mansoni, a widespread trematode parasite that causes the human disease schistosomiasis. According to phylogenetic analysis, S. mansoni invaded the new world during the last 500 years. We hypothesized that new world strains of S. mansoni would have more copies of TEs than old world strains due to the physiological stress associated with invasion of the new world. We quantified the copy number of six TEs (Saci-1, Saci-2 and Saci-3, Perere-1, Merlin-sm1, and SmTRC1) in the genome and the transcriptome of old world and new world strains of S. mansoni, using qPCR relative quantification. As predicted, the genomes of new world parasites contain significantly more copies of class I and class II TEs in both laboratory and field strains. However, such differences are not observed in the transcriptome suggesting that either TE silencing mechanisms have reactivated to control the expression of these elements or the presence of inactive truncated copies of TEs.

  15. Hybridism between Biomphalaria cousini and Biomphalaria amazonica and its susceptibility to Schistosoma mansoni

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    Tatiana Maria Teodoro

    2011-11-01

    Full Text Available Molecular techniques can aid in the classification of Biomphalaria species because morphological differentiation between these species is difficult. Previous studies using phylogeny, morphological and molecular taxonomy showed that some populations studied were Biomphalaria cousini instead of Biomphalaria amazonica. Three different molecular profiles were observed that enabled the separation of B. amazonica from B. cousini. The third profile showed an association between the two and suggested the possibility of hybrids between them. Therefore, the aim of this work was to investigate the hybridism between B. cousini and B. amazonica and to verify if the hybrids are susceptible to Schistosoma mansoni. Crosses using the albinism factor as a genetic marker were performed, with pigmented B. cousini and albino B. amazonica snails identified by polymerase chain reaction-restriction fragment length polymorphism. This procedure was conducted using B. cousini and B. amazonica of the type locality accordingly to Paraense, 1966. In addition, susceptibility studies were performed using snails obtained from the crosses (hybrids and three S. mansoni strains (LE, SJ, AL. The crosses between B. amazonica and B. cousini confirmed the occurrence of hybrids. Moreover, hybrids can be considered potential hosts of S. mansoni because they are susceptible to LE, SJ and AL strains (4.4%, 5.6% and 2.2%, respectively. These results indicate that there is a risk of introducing schistosomiasis mansoni into new areas.

  16. Therapeutic Effects of Allium sativum and Allium cepa in Schistosoma mansoni experimental infection

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    Mona Mohamed Mantawy

    2011-06-01

    Full Text Available The effects of both garlic (Allium sativum and onion (Allium cepa on some biochemical parameters in Schistosoma mansoni infected mice individually and mixed either with or without the currently used drug, praziquantel (PZQ were investigated. These involved some immunological parameters, namely IgM, IgG, interleukins 2 and 6 (IL-2 and 6 and tumor necrosis factor (TNF-α, some antioxidant enzymes [catalase, superoxide dismutase (SOD and glutathione peroxidase (GPX]. In addition, parasitological and histopathological investigations were performed. No changes were observed in the normal control mice treated with dry extract of onion or garlic, individually or mixed, with or without PZQ, compared to the normal healthy control group. Infection with S. mansoni showed an increase in IgG, IgM, IL-2, IL-6, TNF-α and catalase enzyme, accompanied with a decrease in GPX and SOD antioxidant enzyme activities. Remarkable amelioration was noticed in the levels of all the measured parameters in S. mansoni infected mice after administration of the studied extracts. Moreover a significant reduction in worm burden, hepatic and intestinal eggs and oogram count was noticed which was reflected in normalization of liver architecture.

  17. Aristolochia gehrtiiinhibits liver toxicity and apoptosis inSchistosoma malayensis infection

    Institute of Scientific and Technical Information of China (English)

    Khaled M M Koriem; Razmy E Shahabudin; Rafiq Z Jamaludin

    2014-01-01

    Objective:To evaluate a protective effect ofAristolochia gehrtii(A. gehrtii) leaves to inhibit liver toxicity and apoptosis inSchistosoma malayensis(S. malayensis) infection.Methods:Forty male albino mice were divided into four equal groups: group1 control including non-infected healthy mice and groups2,3 &4 subcutaneously infected withS. malayensis cercariae where groups3 &4 pretreated withA. gehrtii leaves(200 mg/kg, bwt) & cinnamoylamide(250 mg/kg, bwt), respectively.Results:S. malayensis caused a significant increase in serumAST, ALT,ALP,MDA,NO, bilirubin, urea, creatinine, total cholesterol,LDL, triglycerides, andHDL levels.The pretreatment ofA. gehrtii leaves and cinnamoylamide significantly inhibited that increase.On the other hand,S. malayensis induced a significant decrease in serum total protein, albumin, globulin, albumin/globulin ratio, bloodSOD andGPx, whileA. gehrtii leaves and cinnamoylamide pretreatment increased the above parameters.Treatment withA. gehrtii leaves and cinnamoylamide toS. malayensis infected mice increased p53 expression but decreased bcl-2 expression.These results were supported by histopathological investigations.Conclusions:A. gehrtii inhibits liver toxicity and apoptosis inS. malayensis infection and this effect is associated with the major cinnamoylamide ingredient ofA. gehrtii leaves.

  18. Schistosomicidal Activity of the Essential Oil of Ageratum conyzoides L. (Asteraceae against Adult Schistosoma mansoni Worms

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    Wilson R. Cunha

    2011-01-01

    Full Text Available The in vitro schistosomicidal effects of the essential oil of Ageratum conyzoides L. (Ac-EO against adult worms of Schistosoma mansoni is reported in this paper. Concerning this activity, Ac-EO was considered to be active, but less effective than the positive control (praziquantel, PZQ in terms of separation of coupled pairs, mortality, decrease in motor activity, and tegumental alterations. However, Ac-EO caused an interesting dose-dependent reduction in the number of eggs of S. mansoni. Precocene I (74.30% and (E-caryophyllene (14.23% were identified as the two major constituents of Ac-EO. These compounds were tested individually and were found to be much less effective than Ac-EO and PZQ. A mixture of the two major compounds in a ratio similar to that found in the Ac-EO was also less effective than Ac-EO, thus revealing that there are no synergistic effects between these components. These results suggest that the essential oil of A. conyzoides is very promising for the development of new schistosomicidal agents.

  19. Morphological Study of Adult Male Worms of Schistosoma mansoni Sambon, 1907 by Scanning Electron Microscopy

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    José Roberto Machado-Silva

    1997-09-01

    Full Text Available Tubercles, spines and sensory receptors are the most studied structures of adult male worms of Schistosoma mansoni isolated in other countries. The purpose of this investigation was to properly define these structures in Brazilian worms. Specimens 7-8 weeks after infection were recovered from albino SW mice and from a wild rodent (Nectomys squamipes and processed for scanning electron microscopy studies. Photomicrographs of the anterior region with the aspects related to the outer and inner regions of both suckers were considered. The ventral portion of the middle region was represented by the anterior of gynaecophoric canal while the dorsal surface was studied in its ventral and dorsal regions mainly focusing the aspect of the tubercles, spines and sensorial papillae. The outer surface of the oral sucker is spiny and spines are bigger, sharp with sensory receptors in their posterior edge. Tubercles with spines or receptors are more concentrated in the middle region and in one of the margins of the gynaecophoric canal. An excretory pore-like structure in the posterior portion was observed. The gynaecophoric canal has few sensory structures, spines broadned in their mid-region and are sharp pointed at the distal end. It was concluded that the presently studied characters are similar to those previously reported

  20. Schistosoma mansoni Sambon, 1907: morphometric differences between adult worms from sympatric rodent and human isolates

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    Neves Renata Heisler

    1998-01-01

    Full Text Available A computer software for image analysis (IMAGE PRO PLUS, MEDIA CYBERNETICS was utilized in male and females adult worms, aiming the morphological characterization of Schistosoma mansoni samples isolated from a slyvatic rodent, Nectomys squamipes, and humans in Sumidouro, Rio de Janeiro, Brazil and recovered from Mus musculus C3H/He. The following characters for males's testicular lobes were analyzed: number, area, density, larger and smaller diameter, longer and shorter axis and perimeter and extension; for females: area, longer and shorter axis, larger and smaller diameter and perimeter of the eggs and spine; oral and ventral suckers area and distance between them in both sex were determined. By the analysis of variance (one way ANOVA significant differences (p<0.05 were observed in all studied characters, except for the density of testicular lobes. Significant differences (p<0.05 were detected for all characters in the female worms. Data ratify that sympatric isolates present phenotypic differences and the adult female characters are useful for the proper identification of S. mansoni isolates.

  1. Host-induced morphological changes of Schistosoma mansoni Sambon, 1907 male worms

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    José Roberto Machado-Silva

    1994-09-01

    Full Text Available In order to evaluate the permissiveness of Nectomys squamipes to Schistosoma mansoni and the influence of the albino mice on the morphological aspects of adult worms derived from a population isolated from N. squamipes, the morphology of adult S. mansoni Sambon, 1907 male worms was studied using a digital image analyser (MOP VIDEOPLAN and light microscopy. Their sources were as follows: (1 recovered from the wild rodent N. squamipes Brants naturally infected from Sumidouro, RJ, Brazil; (2 recovered from albino mice experimentally infected with the strain derived from N. squamipes; (3 recovered after the isolation of a strain derived from aboriginal human infections in Sumidouro. Worms recovered from N. squamipes (group 1 showed body lenght and distance between suckers significantly bigger than those of the specimens maintained in mice (groups 2 and 3. The number of tests in group 1 was statistically less than of groups 2 and 3. Group 2 strains which were maintained in mice, presented the lenght of the worms as the only significant different character. Data show that: (1 N. squamipes is a more suitable host for the development of S. mansoni when compared to the albino mice; (2 a strain of S. mansoni isolated from a natural host undergoes morphological changes after its passage in the white mouse.

  2. Morphometric study of Schistosoma mansoni adult worms recovered from undernourished infected mice

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    Sheilla A Oliveira

    2003-07-01

    Full Text Available Some unfavourable effects of malnutrition of the host on Schistosoma mansoni worm biology and structure have been reported based upon brigthfield microscopy. This paper aims to study by morphometric techniques, some morphological parameters in male and female adult worms recovered from undernourished albino mice in comparison with parasites recovered from well-fed infected mice. Undernourished animals were fed a multideficient and essentially low protein diet (RBD diet and compared to well-fed control mice fed with the commercial diet NUVILAB. Seventy-five days post-infection with 80 cercarie (BL strain animals were sacrificed. All adult worms were fixed in 10% formalin and stained with carmine chloride. One hundred male and 60 female specimens from each group (undernourished and control were examined using an image system analysis Leica Quantimet 500C and the Sigma Scan Measurement System. The following morphometrical parameters were studied: body length and width, oral and ventral suckers, number and area of testicular lobes, length and width of ovary and uterine egg. For statistical analysis, the Student's t test for unpaired samples was applied. Significant differences (p < 0.05 were detected in body length and width, in parameters of suckers, uterine egg width, ovary length and area of testicular lobes, with lower values for specimens from undernourished mice. The nutritional status of the host has negative influence on S. mansoni adult worms, probably through unavailability of essential nutrients to the parasites.

  3. Morphological study of adult male worms of Schistosoma mansoni Sambon, 1907 by confocal laser scanning microscopy

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    Machado-Silva José Roberto

    1998-01-01

    Full Text Available Aiming to detail data obtained through brightfield microscopy (BM on reproductive, excretory and digestive system, specimens of Schistosoma mansoni eight weeks old, were recovered from SW mice, stained with Langeron's carmine and analyzed under a confocal laser scanning microscope CLSM 410 (Carl Zeiss. The reproductive system presented a single and lobate testis, with intercommunications between the lobes without efferent duct. Supernumerary testicular lobe was amorphous and isolated from the normal ones. Collecting tubules (excretory ducts, followed by the excretory bladder, opening to the external media through the excretory pore, were observed at the posterior extremity of the body. In the digestive tract, a cecal swelling was noted at the junction that originates the single cecum. It was concluded that through confocal laser scanning microscopy, new interpretations of morphological structures of S. mansoni worms could be achieved, modifying adopted and current descriptions. The gonad consists of a single lobed testis, similar to that observed in some trematode species. Moreover, the same specimens can be observed either by BM or CLSM, considering that the latter causes only focal and limited damage in tissue structures.

  4. Development of Schistosoma mansoni worms in mice analyzed by bright field and confocal microscopy

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    Carla de Lamare Biolchini

    2006-10-01

    Full Text Available The blood flukes of mammals (Digenea: Schistosomatidae are among trematodes unique whose adult worms have separeted sexes which are dissimilar in appearance. The developmental features, growth and organogenesis of Schistosoma mansoni were studied in Swiss Webster mice by a digital system for image analysis and confocal microscopy. Data so far obtained showed two phases with significative morphological changes at 3-4 weeks post-infection, and a gradual similar development onwards in the reproductive system and tegument. Our male-dependent phase demonstrated that mating occurs before sexual maturing. At week three, the majority of male worms (59% had formed the gynaecophoric canal although testicular lobes and tegumental tubercles were absent. By this time, 33% females had an incipient ovary (without cellular differentiation. At week four, 77.2% males presented testicular lobes with few germinative cells while 26% had developing tegumental tubercles. The immature ovary was observed in 69% females. Suckers followed different pattern of growth between male and females. The size of oral and ventral suckers from six-week-old male worms grew abruptly (3.0 fold more than that of three-week-old. In female worms, maximum growth was attained at week four, reducing in size thereafter. From sixth week onwards, all specimens showed the fully developed reproductive system. Probably, these features are morphological traits which schistosome has experienced from hermaphrodite to dioecy.

  5. Parasite glycans and antibody-mediated immune responses in Schistosoma infection.

    Science.gov (United States)

    van Diepen, Angela; Van der Velden, Niels S J; Smit, Cornelis H; Meevissen, Moniek H J; Hokke, Cornelis H

    2012-08-01

    Schistosome infections in humans are characterized by the development of chronic disease and high re-infection rates after treatment due to the slow development of immunity. It appears that anti-schistosome antibodies are at least partially mediating protective mechanisms. Efforts to develop a vaccine based on immunization with surface-exposed or secreted larval or worm proteins are ongoing. Schistosomes also express a large number of glycans as part of their glycoprotein and glycolipid repertoire, and antibody responses to those glycans are mounted by the infected host. This observation raises the question if glycans might also form novel vaccine targets for immune intervention in schistosomiasis. This review summarizes current knowledge of antibody responses and immunity in experimental and natural infections with Schistosoma, the expression profiles of schistosome glycans (the glycome), and antibody responses to individual antigenic glycan motifs. Future directions to study anti-glycan responses in schistosomiasis in more detail in order to address more precisely the possible role of glycans in antibody-mediated immunity are discussed.

  6. Haemoproteus and Schistosoma synthesize heme polymers similar to Plasmodium hemozoin and beta-hematin.

    Science.gov (United States)

    Chen, M M; Shi, L; Sullivan, D J

    2001-03-01

    Many parasites digest hemoglobin as an amino acid source, but only a few produce heme polymer pigment instead of catabolizing heme via heme oxygenase. This work compares purified heme polymers produced by Haemoproteus columbae and Schistosoma mansoni to that of Plasmodium falciparum hemozoin and synthetic beta-hematin. Fourier-transform infrared spectroscopy identifies the signature peaks of the common iron-carboxylate bond characteristic in all four heme polymers. However, all pigments could be distinguished by quite different three-dimensional structure visualized by Field Emission Inlens Scanning Electron Microscopy. Both P. falciparum and H. columbae heme polymers had a symmetrical shape unlike the amorphous S. mansoni heme polymer and beta-hematin. All four heme pigments serve as templates for heme polymer extension, which was inhibitable by chloroquine and other quinoline antimalarials. The polymers showed different levels of resistance to hydrogen peroxide degradation. This work identifies another genus, Haemoproteus, capable of intracellular heme polymer formation. The different three-dimensional structures of each pigment implicate genus specific formation of heme polymer, variation of inhibition of polymer extension by the quinolines and degradation by hydrogen peroxide.

  7. [Schistosoma haematobium among immigrants consulting in municipal free clinics in Paris in 2003].

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    Deniaud, F; Collignon, A; Guesnon, M S; Squinazi, F; Rouvier, J; Derouineau, J; Ayache, B

    2006-05-01

    The "Consultations de Diagnostic et d'Orientation" (CDO) are free medical consultations for precarious populations, proposed by the Health Department of Paris. More than two-thirds of the CDO patients come from Sub-Saharan Africa. Schistosoma haematobium (SH) is one of the most frequent infectious diseases detected within CDO. More than a thousand people have consulted for the first time in CDO in 2003 in one of the municipal free Clinics which proposes this service. Parasitologic test of urine has been performed among 220 patients and found 24 positive results: viable eggs of SH (10.8%). All 24 patients are male, most of them are under 35 years-old and come from the region of the Senegal River that lies in the junction of Mali, Senegal and Mauritania. We want to remind physicians in non-tropical setting to think of SH when they see a patient originating from Africa. To ask him if he presents haematuria and if not, to prescribe a parasitologic test of urine. If all patients from endemic regions had undergone this screening in 2003, we would have detected about 20 more cases of SH. If treated early enough, it could avoid severe uronephrological complications, which are rare but represent a high health care cost (bladder tumor, renal failure).

  8. Detection of Urinary Tract Pathology in Some Schistosoma haematobium Infected Nigerian Adults

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    O. S. Onile

    2016-01-01

    Full Text Available Screening for Schistosoma haematobium infection and its possible morbidity was carried out in 257 adult participants in Eggua community, Ogun State, Nigeria. Parasitological assessment for the presence of ova of S. haematobium in urine and abdominopelvic ultrasonographic examination for bladder and secondary kidney pathology were carried out. S. haematobium prevalence of 25.68% (66/257 was recorded among the participants. There was a significantly higher prevalence of 69.2% of urinary schistosomiasis in the females than the prevalence of 31.8% in males (P=0.902. The intensity of infections was mostly light (55 (21.8% compared to heavy (10 (3.9% with the mean intensity of 16.7 eggs/10 mL urine. Structural bladder pathology prevalence among participants was 33.9%. The bladder and kidney pathologies observed by ultrasound in subjects with S. haematobium infections included abnormal bladder wall thickness (59%, abnormal bladder shape (15.2%, bladder wall irregularities (15.2%, bladder masses (1.5%, bladder calcification (1.5%, and hydronephrosis (3%. Infection with S. haematobium was associated with bladder pathology. Higher frequencies of bladder abnormalities were observed more in the participants with light intensity of S. haematobium infection than in those with heavy infection. More bladder pathology was also seen in women than in men, although this was not statistically significant. In conclusion, there is evidence that the development of bladder pathology may be associated with S. haematobium infection.

  9. Factors Involved in Schistosoma mansoni Infection, in Rural Areas of Northeast Brazil

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    Coutinho Eridan M

    1997-01-01

    Full Text Available Two contiguous villages in Tracunhaém county (State of Pernambuco, endemic for schistosomiasis, were studied: Itapinassu (138 inhabitants and São Joaquim (91 inhabitants. Agriculture predominates in the former region while ceramics is the main activity in the latter. Although no statistical difference was found regarding prevalence, severe infection (>400 epg predominated in Itapinassu, probably related to the kind of occupation. No association was found between parasite burden and severity of disease, in spite of the high infection rates for Schistosoma mansoni in both communities (approx. 60%. Typical epidemiological features of schistosomiasis such as age-related prevalences and intensities of infection (high in children, low in adults were also mutual characteristics. Nutritional status determined through anthropometric evaluation was carried out by measuring specific anthropometric indicators. A deficit of energy intake, as well as vitamin A and riboflavin deficien