VP Market spreads its wings further

Index Scriptorium Estoniae

2004-01-01

Jaekaubandusettevõte VP Market süüdistab ehituskaupade firmat Senukai tarnijatele surve avaldamises mitte müüa oma tooteid Ermitazas'es, mis on VP Marketi poolt asutatud ehitus- ja majapidamiskaupade poe kett. VP Market'i Eesti esindaja sõnul kaalub ettevõte mõne kohaliku keti ostmist

White spot syndrome virus envelope protein VP28 is involved in the systemic infection of shrimp

NARCIS (Netherlands)

Hulten, van M.C.W.; Witteveldt, J.; Snippe, M.; Vlak, J.M.

2001-01-01

White spot syndrome virus (WSSV) is a large DNA virus infecting shrimp and other crustaceans. The virus particles contain at least five major virion proteins, of which three (VP26, VP24, and VP15) are present in the rod-shaped nucleocapsid and two (VP28 and VP19) reside in the envelope. The mode of

Expression and immunogenicity of novel subunit enterovirus 71 VP1 antigens

International Nuclear Information System (INIS)

Xu, Juan; Wang, Shixia; Gan, Weihua; Zhang, Wenhong; Ju, Liwen; Huang, Zuhu; Lu, Shan

2012-01-01

Highlights: ► EV71 is a major emerging infectious disease in many Asian countries. ► Inactivated EV71 vaccines are in clinical studies but their safety and efficacy are unknown. ► Developing subunit based EV71 vaccines is significant and novel antigen design is needed. ► DNA immunization is an efficient tool to test the immunogenicity of VP1 based EV71 vaccines. ► Multiple VP1 antigens are developed showing immunogenic potential. -- Abstract: Hand, foot, and mouth disease (HFMD) is a common viral illness in young children. HFMD is caused by viruses belonging to the enterovirus genus of the picornavirus family. Recently, enterovirus 71 (EV71) has emerged as a virulent agent for HFMD with severe clinical outcomes. In the current report, we conducted a pilot antigen engineering study to optimize the expression and immunogenicity of subunit VP1 antigen for the design of EV71 vaccines. DNA immunization was adopted as a simple technical approach to test different designs of VP1 antigens without the need to express VP1 protein in vitro first. Our studies indicated that the expression and immunogenicity of VP1 protein can be improved with alternated VP1 antigen designs. Data presented in the current report revealed novel pathways to optimize the design of VP1 antigen-based EV71 vaccines.

[Escherichia coli heat-labile enterotoxin B subunit enhances the immune response against canine parvovirus VP2 in mice immunized by VP2 DNA vaccine].

Science.gov (United States)

Han, Dongmei; Zhong, Fei; Li, Xiujin; Wang, Wei; Wang, Xingxing; Pan, Sumin

2011-01-01

To investigate the effect of Escherichia coli heat-labile enterotoxin (LT) B subunit (LTB) gene on canine parvovirus (CPV) VP2 gene vaccine. The LTB gene was amplified by PCR from genomic DNA of E. coli 44815 strain. The VP2-70 fragment (210 bp) encoding major epitope of VP2 (70 amino acids) was amplified by PCR from a plasmid encoding VP2 gene. VP2-70 and LTB genes were inserted into the eukaryotic vector to construct VP2-70 gene,LTB gene and VP2-70-LTB fused gene vectors. The mice were immunized with VP2-70 vector, VP2-70-LTB fused vector, or VP2-70 vector plus LTB vector, respectively. The antibody titers at the different time were measured by using ELISA method. The spleen lymphocyte proliferation activity was analyzed by 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. The sequence of VP2-70 and LTB genes was identified. The recombinant VP2-70 and LTB proteins could be expressed in HEK293T cells in a secretory manner. The mice immunized with VP2-70 vector, VP2-70-LTB vector or VP2-70 vector plus LTB vector could generate the specific antibody against VP2 protein. The antibody titer immunized with VP2-70-LTB vector reached 1:5120 at 35 d post immunization, significantly higher than that of other two groups (P vaccine in mice.

Dynamic Phosphorylation of VP30 Is Essential for Ebola Virus Life Cycle.

Science.gov (United States)

Biedenkopf, Nadine; Lier, Clemens; Becker, Stephan

2016-05-15

the situation is more complex and that primary transcription as well as the rescue of recombinant Ebola virus also requires the transient phosphorylation of VP30. VP30 encodes six N-proximal serine residues that serve as phosphorylation acceptor sites. The present study shows that the dynamic phosphorylation of serine at position 29 alone is sufficient to activate primary viral transcription. Our results indicate a series of phosphorylation/dephosphorylation events that trigger binding to and release from the nucleocapsid and transcription complex to be essential for the full activity of VP30. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

Expression and immunogenicity of novel subunit enterovirus 71 VP1 antigens

Energy Technology Data Exchange (ETDEWEB)

Xu, Juan [China-US Vaccine Research Center, The First Affiliated Hospital, Nanjing Medical University (China); Department of Microbiology and Immunology, Nanjing Medical University (China); Wang, Shixia [China-US Vaccine Research Center, The First Affiliated Hospital, Nanjing Medical University (China); Department of Medicine, University of Massachusetts Medical School (United States); Gan, Weihua [Department of Pediatrics, The Second Affiliated Hospital, Nanjing Medical University (China); Zhang, Wenhong [Department of Infectious Diseases, Huashan Hospital, Fudan University (China); Ju, Liwen [School of Public Health, Fudan University (China); Huang, Zuhu [Department of Infectious Diseases, The First Affiliated Hospital, Nanjing Medical University (China); China-US Vaccine Research Center, The First Affiliated Hospital, Nanjing Medical University (China); Lu, Shan, E-mail: shan.lu@umassmed.edu [Department of Infectious Diseases, The First Affiliated Hospital, Nanjing Medical University (China); China-US Vaccine Research Center, The First Affiliated Hospital, Nanjing Medical University (China); Department of Medicine, University of Massachusetts Medical School (United States)

2012-04-20

Highlights: Black-Right-Pointing-Pointer EV71 is a major emerging infectious disease in many Asian countries. Black-Right-Pointing-Pointer Inactivated EV71 vaccines are in clinical studies but their safety and efficacy are unknown. Black-Right-Pointing-Pointer Developing subunit based EV71 vaccines is significant and novel antigen design is needed. Black-Right-Pointing-Pointer DNA immunization is an efficient tool to test the immunogenicity of VP1 based EV71 vaccines. Black-Right-Pointing-Pointer Multiple VP1 antigens are developed showing immunogenic potential. -- Abstract: Hand, foot, and mouth disease (HFMD) is a common viral illness in young children. HFMD is caused by viruses belonging to the enterovirus genus of the picornavirus family. Recently, enterovirus 71 (EV71) has emerged as a virulent agent for HFMD with severe clinical outcomes. In the current report, we conducted a pilot antigen engineering study to optimize the expression and immunogenicity of subunit VP1 antigen for the design of EV71 vaccines. DNA immunization was adopted as a simple technical approach to test different designs of VP1 antigens without the need to express VP1 protein in vitro first. Our studies indicated that the expression and immunogenicity of VP1 protein can be improved with alternated VP1 antigen designs. Data presented in the current report revealed novel pathways to optimize the design of VP1 antigen-based EV71 vaccines.

Structures of Rotavirus Reassortants Demonstrate Correlation of Altered Conformation of the VP4 Spike and Expression of Unexpected VP4-Associated Phenotypes

Science.gov (United States)

Pesavento, Joseph B.; Billingsley, Angela M.; Roberts, Ed J.; Ramig, Robert F.; Prasad, B. V. Venkataram

2003-01-01

Numerous prior studies have indicated that viable rotavirus reassortants containing structural proteins of heterologous parental origin may express unexpected phenotypes, such as changes in infectivity and immunogenicity. To provide a structural basis for alterations in phenotypic expression, a three-dimensional structural analysis of these reassortants was conducted. The structures of the reassortants show that while VP4 generally maintains the parental structure when moved to a heterologous protein background, in certain reassortants, there are subtle alterations in the conformation of VP4. The alterations in VP4 conformation correlated with expression of unexpected VP4-associated phenotypes. Interactions between heterologous VP4 and VP7 in reassortants expressing unexpected phenotypes appeared to induce the conformational alterations seen in VP4. PMID:12584352

VP7: an attachment protein of bluetongue virus for cellular receptors in Culicoides variipennis.

Science.gov (United States)

Xu, G; Wilson, W; Mecham, J; Murphy, K; Zhou, E M; Tabachnick, W

1997-07-01

The importance of VP7 of bluetongue virus (BTV) in the binding of BTV to membrane proteins of the BTV vector Culicoides variipennis was investigated. Core BTV particles, prepared from whole viruses, lacked outer proteins VP2 and VP5 and had VP7 exposed. More core particles and whole viruses bound to membrane preparations of adults of C. variipennis and KC cells, which were cultured from this vector insect, than to membrane preparations of Manduca sexta larvae. More core particles than whole viruses bound to membrane preparations of adults of C. variipennis and KC cells. Polyclonal anti-idiotypic antibodies (anti-Id), which were made against an antigen-combining region of an anti-BTV-10 VP7 antibody and functionally mimicked VP7, bound more to the membrane preparations of adults of C. variipennis and KC cells, and less to cytosol preparations. In Western overalay analysis, the Culicoides plasma membrane preparation reduced binding of an anti-VP7 monoclonal antibody to VP7. Whole and core BTV particles and the anti-Id bound to a membrane protein with a molecular mass of 23 kDa that was present predominantly in membrane preparations of adults of C. variipennis and KC cells. This protein was present in much lower concentrations in membrane preparations of C6/36 and DM-2 insect cells.

Comparing Enterovirus 71 with Coxsackievirus A16 by analyzing nucleotide sequences and antigenicity of recombinant proteins of VP1s and VP4s

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Sun Yu

2011-11-01

Full Text Available Abstract Background Enterovirus 71 (EV71 and Coxsackievirus A16 (CA16 are two major etiological agents of Hand, Foot and Mouth Disease (HFMD. EV71 is associated with severe cases but not CA16. The mechanisms contributed to the different pathogenesis of these two viruses are unknown. VP1 and VP4 are two major structural proteins of these viruses, and should be paid close attention to. Results The sequences of vp1s from 14 EV71 and 14 CA16, and vp4s from 10 EV71 and 1 CA16 isolated in this study during 2007 to 2009 HFMD seasons were analyzed together with the corresponding sequences available in GenBank using DNAStar and MEGA 4.0. Phylogenetic analysis of complete vp1s or vp4s showed that EV71 isolated in Beijing belonged to C4 and CA16 belonged to lineage B2 (lineage C. VP1s and VP4s from 4 strains of viruses expressed in E. coli BL21 cells were used to detect IgM and IgG in human sera by Western Blot. The detection of IgM against VP1s of EV71 and CA16 showed consistent results with current infection, while none of the sera were positive against VP4s of EV71 and CA16. There was significant difference in the positive rates between EV71 VP1 and CA16 VP1 (χ2 = 5.02, P 2 = 15.30, P 2 = 26.47, P 2 = 16.78, P Conclusions EV71 and CA16 were highly diverse in the nucleotide sequences of vp1s and vp4s. The sera positive rates of VP1 and VP4 of EV71 were lower than those of CA16 respectively, which suggested a less exposure rate to EV71 than CA16 in Beijing population. Human serum antibodies detected by Western blot using VP1s and VP4s as antigen indicated that the immunological reaction to VP1 and VP4 of both EV71 and CA16 was different.

Magma intrusion in the upper crust of Abu Dabbab area, South East of Egypt from Vp and Vp/Vs tomography

International Nuclear Information System (INIS)

Hosny, A.; El Hady, S.M.; Mohamed, A.A.; Panza, G.F.

2007-12-01

3-D images of P-wave velocity and Vp/Vs ratio have been produced for the upper crust of the Abu Dabbab area, North Mars Alam city. The inversion of local travel times of high quality data recorded at eleven mobile seismic stations around the study area is carried out. The best, in the least-squares sense, 1-D Vp model and the average value of Vp/Vs (1.72) were computed as prerequisites of the 3-D inversion that reaches a depth of 14 km. From the 3-D model it is evident that the distributions of Vp and Vp/Vs are characterized by marked lateral and vertical variations delineating structural heterogeneities. Due to the presence of a thin layer of sedimentary rocks saturated with surface water, low P-wave velocity and high Vp/Vs values are noticed near the surface. At greater depths, high Vp and low Vp/Vs zones may indicate crustal rocks with relatively higher rigidity and brittle behavior, while high Vp/Vs and low Vp may identify zones of relatively softer rocks, with ductile behavior. Low P-wave velocity values are observed at the intersections among the faults. Some magma intrusions could be associated to the Vp/Vs values which form an elongated anomaly, in the western part of the study area, which extends from a depth of 12 km to about 1-2 km of depth. If the obtained 3-D model is used in the relocation of selected events, they turn out to be strongly clustered in correspondence with the high velocity anomalies detected in the central part of the study area. Most of the seismicity tends to occur at the boundaries between the high and low velocity anomalies and at pre-existing weakness zones, i.e. the areas of intersection among different faults. The occurrence of the seismic activity in the vicinity of low velocity anomalies and at the boundary between velocity contrast could also be explained by the occurrence of serpentinization processes in the crust of the study area. (author)

High-resolution Crystal Structure of Dimeric VP40 From Sudan ebolavirus.

Science.gov (United States)

Clifton, Matthew C; Bruhn, Jessica F; Atkins, Kateri; Webb, Terry L; Baydo, Ruth O; Raymond, Amy; Lorimer, Donald D; Edwards, Thomas E; Myler, Peter J; Saphire, Erica Ollmann

2015-10-01

Ebolaviruses cause severe hemorrhagic fever. Central to the Ebola life cycle is the matrix protein VP40, which oligomerizes and drives viral budding. Here we present the crystal structure of the Sudan virus (SUDV) matrix protein. This structure is higher resolution (1.6 Å) than previously achievable. Despite differences in the protein purification, we find that it still forms a stable dimer in solution, as was noted for other Ebola VP40s. Although the N-terminal domain interface by which VP40 dimerizes is conserved between Ebola virus and SUDV, the C-terminal domain interface by which VP40 dimers may further assemble is significantly smaller in this SUDV assembly. © The Author 2015. Published by Oxford University Press on behalf of the Infectious Diseases Society of America. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

Phylogenetic characterization of Canine Parvovirus VP2 partial sequences from symptomatic dogs samples.

Science.gov (United States)

Zienius, D; Lelešius, R; Kavaliauskis, H; Stankevičius, A; Šalomskas, A

2016-01-01

The aim of the present study was to detect canine parvovirus (CPV) from faecal samples of clinically ill domestic dogs by polymerase chain reaction (PCR) followed by VP2 gene partial sequencing and molecular characterization of circulating strains in Lithuania. Eleven clinically and antigen-tested positive dog faecal samples, collected during the period of 2014-2015, were investigated by using PCR. The phylogenetic investigations indicated that the Lithuanian CPV VP2 partial sequences (3025-3706 cds) were closely related and showed 99.0-99.9% identity. All Lithuanian sequences were associated with one phylogroup, but grouped in different clusters. Ten of investigated Lithuanian CPV VP2 sequences were closely associated with CPV 2a antigenic variant (99.4% nt identity). Five CPV VP2 sequences from Lithuania were related to CPV-2a, but were rather divergent (6.8 nt differences). Only one CPV VP2 sequence from Lithuania was associated (99.3% nt identity) with CPV-2b VP2 sequences from France, Italy, USA and Korea. The four of eleven investigated Lithuanian dogs with CPV infection symptoms were vaccinated with CPV-2 vaccine, but their VP2 sequences were phylogenetically distantly associated with CPV vaccine strains VP2 sequences (11.5-15.8 nt differences). Ten Lithuanian CPV VP2 sequences had monophyletic relations among the close geographically associated samples, but five of them were rather divergent (1.0% less sequence similarity). The one Lithuanian CPV VP2 sequence was closely related with CPV-2b antigenic variant. All the Lithuanian CPV VP2 partial sequences were conservative and phylogenetically low associated with most commonly used CPV vaccine strains.

Image quality and radiation dose of brain computed tomography in children: effects of decreasing tube voltage from 120 kVp to 80 kVp

International Nuclear Information System (INIS)

Park, Ji Eun; Choi, Young Hun; Cheon, Jung-Eun; Kim, Woo Sun; Kim, In-One; Cho, Hyun Suk; Ryu, Young Jin; Kim, Yu Jin

2017-01-01

Computed tomography (CT) has generated public concern associated with radiation exposure, especially for children. Lowering the tube voltage is one strategy to reduce radiation dose. To assess the image quality and radiation dose of non-enhanced brain CT scans acquired at 80 kilo-voltage peak (kVp) compared to those at 120 kVp in children. Thirty children who had undergone both 80- and 120-kVp non-enhanced brain CT were enrolled. For quantitative analysis, the mean attenuation of white and gray matter, attenuation difference, noise, signal-to-noise ratio, contrast-to-noise ratio and posterior fossa artifact index were measured. For qualitative analysis, noise, gray-white matter differentiation, artifact and overall image quality were scored. Radiation doses were evaluated by CT dose index, dose-length product and effective dose. The mean attenuations of gray and white matter and contrast-to-noise ratio were significantly increased at 80 kVp, while parameters related to image noise, i.e. noise, signal-to-noise ratio and posterior fossa artifact index were higher at 80 kVp than at 120 kVp. In qualitative analysis, 80-kVp images showed improved gray-white differentiation but more artifacts compared to 120-kVp images. Subjective image noise and overall image quality scores were similar between the two scans. Radiation dose parameters were significantly lower at 80 kVp than at 120 kVp. In pediatric non-enhanced brain CT scans, a decrease in tube voltage from 120 kVp to 80 kVp resulted in improved gray-white matter contrast, comparable image quality and decreased radiation dose. (orig.)

Image quality and radiation dose of brain computed tomography in children: effects of decreasing tube voltage from 120 kVp to 80 kVp.

Science.gov (United States)

Park, Ji Eun; Choi, Young Hun; Cheon, Jung-Eun; Kim, Woo Sun; Kim, In-One; Cho, Hyun Suk; Ryu, Young Jin; Kim, Yu Jin

2017-05-01

Computed tomography (CT) has generated public concern associated with radiation exposure, especially for children. Lowering the tube voltage is one strategy to reduce radiation dose. To assess the image quality and radiation dose of non-enhanced brain CT scans acquired at 80 kilo-voltage peak (kVp) compared to those at 120 kVp in children. Thirty children who had undergone both 80- and 120-kVp non-enhanced brain CT were enrolled. For quantitative analysis, the mean attenuation of white and gray matter, attenuation difference, noise, signal-to-noise ratio, contrast-to-noise ratio and posterior fossa artifact index were measured. For qualitative analysis, noise, gray-white matter differentiation, artifact and overall image quality were scored. Radiation doses were evaluated by CT dose index, dose-length product and effective dose. The mean attenuations of gray and white matter and contrast-to-noise ratio were significantly increased at 80 kVp, while parameters related to image noise, i.e. noise, signal-to-noise ratio and posterior fossa artifact index were higher at 80 kVp than at 120 kVp. In qualitative analysis, 80-kVp images showed improved gray-white differentiation but more artifacts compared to 120-kVp images. Subjective image noise and overall image quality scores were similar between the two scans. Radiation dose parameters were significantly lower at 80 kVp than at 120 kVp. In pediatric non-enhanced brain CT scans, a decrease in tube voltage from 120 kVp to 80 kVp resulted in improved gray-white matter contrast, comparable image quality and decreased radiation dose.

Image quality and radiation dose of brain computed tomography in children: effects of decreasing tube voltage from 120 kVp to 80 kVp

Energy Technology Data Exchange (ETDEWEB)

Park, Ji Eun [Kyung Hee University Hospital, Department of Radiology, Graduate School, Seoul (Korea, Republic of); Choi, Young Hun [Seoul National University Hospital, Department of Radiology, Seoul (Korea, Republic of); Seoul National University College of Medicine, Department of Radiology, Seoul (Korea, Republic of); Cheon, Jung-Eun; Kim, Woo Sun; Kim, In-One [Seoul National University Hospital, Department of Radiology, Seoul (Korea, Republic of); Seoul National University College of Medicine, Department of Radiology, Seoul (Korea, Republic of); Seoul National University Medical Research Center, Institute of Radiation Medicine, Seoul (Korea, Republic of); Cho, Hyun Suk; Ryu, Young Jin; Kim, Yu Jin [Seoul National University Hospital, Department of Radiology, Seoul (Korea, Republic of)

2017-05-15

Computed tomography (CT) has generated public concern associated with radiation exposure, especially for children. Lowering the tube voltage is one strategy to reduce radiation dose. To assess the image quality and radiation dose of non-enhanced brain CT scans acquired at 80 kilo-voltage peak (kVp) compared to those at 120 kVp in children. Thirty children who had undergone both 80- and 120-kVp non-enhanced brain CT were enrolled. For quantitative analysis, the mean attenuation of white and gray matter, attenuation difference, noise, signal-to-noise ratio, contrast-to-noise ratio and posterior fossa artifact index were measured. For qualitative analysis, noise, gray-white matter differentiation, artifact and overall image quality were scored. Radiation doses were evaluated by CT dose index, dose-length product and effective dose. The mean attenuations of gray and white matter and contrast-to-noise ratio were significantly increased at 80 kVp, while parameters related to image noise, i.e. noise, signal-to-noise ratio and posterior fossa artifact index were higher at 80 kVp than at 120 kVp. In qualitative analysis, 80-kVp images showed improved gray-white differentiation but more artifacts compared to 120-kVp images. Subjective image noise and overall image quality scores were similar between the two scans. Radiation dose parameters were significantly lower at 80 kVp than at 120 kVp. In pediatric non-enhanced brain CT scans, a decrease in tube voltage from 120 kVp to 80 kVp resulted in improved gray-white matter contrast, comparable image quality and decreased radiation dose. (orig.)

Ces-VP: consultation expert system for vector programming of nuclear codes

International Nuclear Information System (INIS)

Fujisaki, Masahide; Makino, Mitsuhiro; Ishiguro, Misako

1988-08-01

Ces-VP is a prototype rule-based expert system for consulting the vector programming, based on the knowledge of vectorization of nuclear codes at JAERI during these 10 years. Experts in vectorization can restructure nuclear codes with high performance on vector processors, since they have know-how for choosing the best technique among a lot of techniques that were acquired from the experience of vectorization in the past. Frequency in trial and error will be reduced if a beginner can easily use the know-how of experts. In this report, at first the contents of Ces-VP and its intention are shown. Then, the method for acquiring the know-how of vectorization and the method for making rules from the know-how are described. The outline of Ces-VP implemented on Fujitsu expert tool ESHELL is described. Finally, the availability of Ces-VP is evaluated from the data gathered from practical use and its present problems are discussed. (author)

Rapid detection of EBOLA VP40 in microchip immunofiltration assay

Science.gov (United States)

Miethe, Peter; Gary, Dominik; Hlawatsch, Nadine; Gad, Anne-Marie

2015-05-01

In the spring of 2014, the Ebola virus (EBOV) strain Zaire caused a dramatic outbreak in several regions of West Africa. The RT-PCR and antigen capture diagnostic proved to be effective for detecting EBOV in blood and serum. In this paper, we present data of a rapid antigen capture test for the detection of VP40. The test was performed in a microfluidic chip for immunofiltration analysis. The chip integrates all necessary assay components. The analytical sensitivity of the rapid test was 8 ng/ml for recombinant VP40. In serum and whole blood samples spiked with virus culture material, the detection limit was 2.2 x 102 PFU/ml. The performance data of the rapid test (15 min) are comparable to that of the VP40 laboratory ELISA.

Local licensers and recovering in VP ellipsis

Directory of Open Access Journals (Sweden)

Sonia Cyrino

2005-12-01

Full Text Available The core properties of VP ellipsis in English and Portuguese may be captured assuming that the elliptical constituent is licensed under local c-command by a verbal element in a sentence functional head. However, the lack of VP ellipsis in most Romance languages and in German, despite the existence of verb movement to sentence functional projections in these languages, suggests that a parameter is involved. Empirical evidence indicates that this parametric variation should not be attributed to a specific functional head because the functional head occupied by the verbal licenser may vary across languages and language varieties. So we will claim that the existence vs. absence of VP ellipsis in the languages considered in this study is due to the features of the functional head that intervenes between the verbal licenser and the elliptical vP phase.

Expresión y purificación de las proteínas estructurales del rotavirus VP5* y VP8* en bacterias E. coli BL21(DE3

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Luz Yurany Moreno

2013-01-01

dirigidos contra ellas,  anticipan su utilidad como herramientas  en la caracterización de la interacción virus-célula.  Palabras clave: rotavirus; VP5*; VP8*; proteínas recombinantes; E. coli BL21(DE3 Abstract The characterization of rotavirus structural proteins and the cell surface proteins involved in virion binding and penetration depends on the availability of substantial amounts of highly purified proteins. The aim of the present work was to express and purify the rotavirus structural proteins VP5* and VP8* in order to produce polyclonal antibodies against them. Recombinant proteins VP5* (rVP5* and VP8* (rVP8* were expressed in E. coli cells transfected with pGEX-4T or pET 28a containing their corresponding encoding sequences. Culture medium, bacterial concentration before induction, inductor concentration and induction time were used as variables. The greater proportion of rVP8* was obtained when transfected bacteria were grown in medium LB and induction was started by adding 1 mM IPTG to cells at OD 600 nm 0.5 followed by 6 h-induction. The highest proportion of rVP5* was reached when cells at OD 600 nm 0.2 were induced with 0.5 mM IPTG for 4 h in medium 2XYT containing 2% glucose. The recombinant proteins accumulated in the insoluble fraction were solubilized with ionic and non-ionic detergents, and purified by affinity chromatography before being used as antigens for production of rabbit polyclonal antibodies. The antibodies produced were characterized through their ability to recognize the corresponding antigens in ELISA, Western blotting, and immunochemistry assays in rotavirus infected cells. The amount and purity of recombinant proteins obtained in this work, and the antibodies against them, are expected to be useful for the characterization of the virus-cell interaction. Keywords: rotavirus; VP5*; VP8*; recombinant proteins; E. coli BL21(DE3

Identification of continuous human B-cell epitopes in the VP35, VP40, nucleoprotein and glycoprotein of Ebola virus.

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Pierre Becquart

Full Text Available Ebola virus (EBOV is a highly virulent human pathogen. Recovery of infected patients is associated with efficient EBOV-specific immunoglobulin G (IgG responses, whereas fatal outcome is associated with defective humoral immunity. As B-cell epitopes on EBOV are poorly defined, we sought to identify specific epitopes in four EBOV proteins (Glycoprotein (GP, Nucleoprotein (NP, and matrix Viral Protein (VP40 and VP35. For the first time, we tested EBOV IgG+ sera from asymptomatic individuals and symptomatic Gabonese survivors, collected during the early humoral response (seven days after the end of symptoms and the late memory phase (7-12 years post-infection. We also tested sera from EBOV-seropositive patients who had never had clinical signs of hemorrhagic fever or who lived in non-epidemic areas (asymptomatic subjects. We found that serum from asymptomatic individuals was more strongly reactive to VP40 peptides than to GP, NP or VP35. Interestingly, anti-EBOV IgG from asymptomatic patients targeted three immunodominant regions of VP40 reported to play a crucial role in virus assembly and budding. In contrast, serum from most survivors of the three outbreaks, collected a few days after the end of symptoms, reacted mainly with GP peptides. However, in asymptomatic subjects the longest immunodominant domains were identified in GP, and analysis of the GP crystal structure revealed that these domains covered a larger surface area of the chalice bowl formed by three GP1 subunits. The B-cell epitopes we identified in the EBOV VP35, VP40, NP and GP proteins may represent important tools for understanding the humoral response to this virus and for developing new antibody-based therapeutics or detection methods.

Identification of continuous human B-cell epitopes in the VP35, VP40, nucleoprotein and glycoprotein of Ebola virus.

Science.gov (United States)

Becquart, Pierre; Mahlakõiv, Tanel; Nkoghe, Dieudonné; Leroy, Eric M

2014-01-01

Ebola virus (EBOV) is a highly virulent human pathogen. Recovery of infected patients is associated with efficient EBOV-specific immunoglobulin G (IgG) responses, whereas fatal outcome is associated with defective humoral immunity. As B-cell epitopes on EBOV are poorly defined, we sought to identify specific epitopes in four EBOV proteins (Glycoprotein (GP), Nucleoprotein (NP), and matrix Viral Protein (VP)40 and VP35). For the first time, we tested EBOV IgG+ sera from asymptomatic individuals and symptomatic Gabonese survivors, collected during the early humoral response (seven days after the end of symptoms) and the late memory phase (7-12 years post-infection). We also tested sera from EBOV-seropositive patients who had never had clinical signs of hemorrhagic fever or who lived in non-epidemic areas (asymptomatic subjects). We found that serum from asymptomatic individuals was more strongly reactive to VP40 peptides than to GP, NP or VP35. Interestingly, anti-EBOV IgG from asymptomatic patients targeted three immunodominant regions of VP40 reported to play a crucial role in virus assembly and budding. In contrast, serum from most survivors of the three outbreaks, collected a few days after the end of symptoms, reacted mainly with GP peptides. However, in asymptomatic subjects the longest immunodominant domains were identified in GP, and analysis of the GP crystal structure revealed that these domains covered a larger surface area of the chalice bowl formed by three GP1 subunits. The B-cell epitopes we identified in the EBOV VP35, VP40, NP and GP proteins may represent important tools for understanding the humoral response to this virus and for developing new antibody-based therapeutics or detection methods.

Ebolavirus VP35 uses a bimodal strategy to bind dsRNA for innate immune suppression

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Kimberlin, Christopher R.; Bornholdt, Zachary A.; Li, Sheng; Woods, Jr., Virgil L.; MacRae, Ian J.; Saphire, Erica Ollmann (Scripps); (UCSD)

2010-03-12

Ebolavirus causes a severe hemorrhagic fever and is divided into five distinct species, of which Reston ebolavirus is uniquely nonpathogenic to humans. Disease caused by ebolavirus is marked by early immunosuppression of innate immune signaling events, involving silencing and sequestration of double-stranded RNA (dsRNA) by the viral protein VP35. Here we present unbound and dsRNA-bound crystal structures of the dsRNA-binding domain of Reston ebolavirus VP35. The structures show that VP35 forms an unusual, asymmetric dimer on dsRNA binding, with each of the monomers binding dsRNA in a different way: one binds the backbone whereas the other caps the terminus. Additional SAXS, DXMS, and dsRNA-binding experiments presented here support a model of cooperative dsRNA recognition in which binding of the first monomer assists binding of the next monomer of the oligomeric VP35 protein. This work illustrates how ebolavirus VP35 could mask key recognition sites of molecules such as RIG-I, MDA-5, and Dicer to silence viral dsRNA in infection.

Molecular cloning and sequence analysis of VP6 gene of giant ...

African Journals Online (AJOL)

Rotavirus (family Reoviridae) is the leading cause of severe gastroenteritis in human and animals worldwide. The genome of rotavirus comprises of 11 segments of dsRNA and encodes six structural proteins (VP1 to VP4, VP6 and VP7) and six non structural proteins (NSP1 to NSP6). VP6 is a group of antigen of rotavirus ...

RNA binding specificity of Ebola virus transcription factor VP30.

Science.gov (United States)

Schlereth, Julia; Grünweller, Arnold; Biedenkopf, Nadine; Becker, Stephan; Hartmann, Roland K

2016-09-01

The transcription factor VP30 of the non-segmented RNA negative strand Ebola virus balances viral transcription and replication. Here, we comprehensively studied RNA binding by VP30. Using a novel VP30:RNA electrophoretic mobility shift assay, we tested truncated variants of 2 potential natural RNA substrates of VP30 - the genomic Ebola viral 3'-leader region and its complementary antigenomic counterpart (each ∼155 nt in length) - and a series of other non-viral RNAs. Based on oligonucleotide interference, the major VP30 binding region on the genomic 3'-leader substrate was assigned to the internal expanded single-stranded region (∼ nt 125-80). Best binding to VP30 was obtained with ssRNAs of optimally ∼ 40 nt and mixed base composition; underrepresentation of purines or pyrimidines was tolerated, but homopolymeric sequences impaired binding. A stem-loop structure, particularly at the 3'-end or positioned internally, supports stable binding to VP30. In contrast, dsRNA or RNAs exposing large internal loops flanked by entirely helical arms on both sides are not bound. Introduction of a 5´-Cap(0) structure impaired VP30 binding. Also, ssDNAs bind substantially weaker than isosequential ssRNAs and heparin competes with RNA for binding to VP30, indicating that ribose 2'-hydroxyls and electrostatic contacts of the phosphate groups contribute to the formation of VP30:RNA complexes. Our results indicate a rather relaxed RNA binding specificity of filoviral VP30, which largely differs from that of the functionally related transcription factor of the Paramyxoviridae which binds to ssRNAs as short as 13 nt with a preference for oligo(A) sequences.

In commemoration of professor V.P. Karpov

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Semyonova L.S.

2013-10-01

Full Text Available This article is about professor Karpov V.P., a prominent scientist, first rector of Yekaterinoslav Medical Academy. Biography of a great investigator, his main achievements in the area of histology, biology, theory and history of medicine was studied. Professor Karpov V.P. always combined his great scientific, organizational and research work with social activity. Monographs of professor Karpov V.P. and conferences organized by him were of great importance in the solution of such new problems as theary of microscope and cell amitosis. Professor Karpov is a founder of a large school of histology. Thanks to his active participation and personal guidance, in 1917 department of histology was founded in Yekaterinoslav Medical Institute. The author of the article has analyzed Hippocrates` works translated into Russian by professor Karpov V.P. and pointed out their significance for modern medical science and practice.

Structure of the Ebola VP35 interferon inhibitory domain.

Science.gov (United States)

Leung, Daisy W; Ginder, Nathaniel D; Fulton, D Bruce; Nix, Jay; Basler, Christopher F; Honzatko, Richard B; Amarasinghe, Gaya K

2009-01-13

Ebola viruses (EBOVs) cause rare but highly fatal outbreaks of viral hemorrhagic fever in humans, and approved treatments for these infections are currently lacking. The Ebola VP35 protein is multifunctional, acting as a component of the viral RNA polymerase complex, a viral assembly factor, and an inhibitor of host interferon (IFN) production. Mutation of select basic residues within the C-terminal half of VP35 abrogates its dsRNA-binding activity, impairs VP35-mediated IFN antagonism, and attenuates EBOV growth in vitro and in vivo. Because VP35 contributes to viral escape from host innate immunity and is required for EBOV virulence, understanding the structural basis for VP35 dsRNA binding, which correlates with suppression of IFN activity, is of high importance. Here, we report the structure of the C-terminal VP35 IFN inhibitory domain (IID) solved to a resolution of 1.4 A and show that VP35 IID forms a unique fold. In the structure, we identify 2 basic residue clusters, one of which is important for dsRNA binding. The dsRNA binding cluster is centered on Arg-312, a highly conserved residue required for IFN inhibition. Mutation of residues within this cluster significantly changes the surface electrostatic potential and diminishes dsRNA binding activity. The high-resolution structure and the identification of the conserved dsRNA binding residue cluster provide opportunities for antiviral therapeutic design. Our results suggest a structure-based model for dsRNA-mediated innate immune antagonism by Ebola VP35 and other similarly constructed viral antagonists.

Recombinant VP1 protein as a potential marker for the diagnosis of acute hepatitis A virus infection.

Science.gov (United States)

da Silva Junior, Haroldo Cid; da Silva, Edimilson Domingos; Lewis-Ximenez de Souza Rodrigues, Lia Laura; Medeiros, Marco Alberto

2017-07-01

Since hepatitis A virus (HAV) production is time-consuming and expensive, the use of recombinant proteins may represent an alternative source of antigens for diagnostic purposes. The present study aimed to express, purify and evaluate the potential of recombinant VP1 protein (rVP1) as a marker for the diagnosis of acute HAV infection. The rVP1 was expressed and purified successfully from Escherichia coli. The purified rVP1 was used to establish an in-house enzyme-linked immunosorbent assay (ELISA-rVP1) for detection of IgM antibodies in sera from HAV-positive patients. For a cut-off point of 0.351, the sensitivity and specificity of ELISA-rVP1 were 100.0% and 95.0%, respectively. These results indicate that rVP1 may be a useful antigen for detection of IgM antibodies against HAV. Copyright © 2017 Elsevier B.V. All rights reserved.

Unusual structural transition of antimicrobial VP1 peptide.

Science.gov (United States)

Shanmugam, Ganesh; Phambu, Nsoki; Polavarapu, Prasad L

2011-05-01

VP1 peptide, an active domain of m-calpain enzyme with antimicrobial activity is found to undergo an unusual conformational transition in trifluoroethanol (TFE) solvent. The nature of, and time dependent variations in, circular dichroism associated with the amide I vibrations, suggest that VP1 undergoes self-aggregation forming anti-parallel β-sheet structure in TFE. Transmission electron micrograph (TEM) images revealed that β-sheet aggregates formed by VP1 possess fibril-like assemblies. Copyright © 2011 Elsevier B.V. All rights reserved.

Expression and purification of recombinant polyomavirus VP2 protein and its interactions with polyomavirus proteins

Science.gov (United States)

Cai, X.; Chang, D.; Rottinghaus, S.; Consigli, R. A.; Spooner, B. S. (Principal Investigator)

1994-01-01

Recombinant polyomavirus VP2 protein was expressed in Escherichia coli (RK1448), using the recombinant expression system pFPYV2. Recombinant VP2 was purified to near homogeneity by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, electroelution, and Extracti-Gel chromatography. Polyclonal serum to this protein which reacted specifically with recombinant VP2 as well as polyomavirus virion VP2 and VP3 on Western blots (immunoblots) was produced. Purified VP2 was used to establish an in vitro protein-protein interaction assay with polyomavirus structural proteins and purified recombinant VP1. Recombinant VP2 interacted with recombinant VP1, virion VP1, and the four virion histones. Recombinant VP1 coimmunoprecipitated with recombinant VP2 or truncated VP2 (delta C12VP2), which lacked the carboxy-terminal 12 amino acids. These experiments confirmed the interaction between VP1 and VP2 and revealed that the carboxyterminal 12 amino acids of VP2 and VP3 were not necessary for formation of this interaction. In vivo VP1-VP2 interaction study accomplished by cotransfection of COS-7 cells with VP2 and truncated VP1 (delta N11VP1) lacking the nuclear localization signal demonstrated that VP2 was capable of translocating delta N11VP1 into the nucleus. These studies suggest that complexes of VP1 and VP2 may be formed in the cytoplasm and cotransported to the nucleus for virion assembly to occur.

Oligomerization and polymerization of the filovirus matrix protein VP40

International Nuclear Information System (INIS)

Timmins, Joanna; Schoehn, Guy; Kohlhaas, Christine; Klenk, Hans-Dieter; Ruigrok, Rob W.H.; Weissenhorn, Winfried

2003-01-01

The matrix protein VP40 from Ebola virus plays an important role in the assembly process of virus particles by interacting with cellular factors, cellular membranes, and the ribonuclearprotein particle complex. Here we show that the N-terminal domain of VP40 folds into a mixture of two different oligomeric states in vitro, namely hexameric and octameric ringlike structures, as detected by gel filtration chromatography, chemical cross-linking, and electron microscopy. Octamer formation depends largely on the interaction with nucleic acids, which in turn confers in vitro SDS resistance. Refolding experiments with a nucleic acid free N-terminal domain preparation reveal a mostly dimeric form of VP40, which is transformed into an SDS resistant octamer upon incubation with E. coli nucleic acids. In addition, we demonstrate that the N-terminal domain of Marburg virus VP40 also folds into ringlike structures, similar to Ebola virus VP40. Interestingly, Marburg virus VP40 rings reveal a high tendency to polymerize into rods composed of stacked rings. These results may suggest distinct roles for different oligomeric forms of VP40 in the filovirus life cycle

[Immunoreactivity of chimeric proteins carrying poliovirus epitopes on the VP6 of rotavirus as a vector].

Science.gov (United States)

Pan, X-X; Zhao, B-X; Teng, Y-M; Xia, W-Y; Wang, J; Li, X-F; Liao, G-Y; Yang, С; Chen, Y-D

2016-01-01

Rotavirus and poliovirus continue to present significant risks and burden of disease to children in developing countries. Developing a combined vaccine may effectively prevent both illnesses and may be advantageous in terms of maximizing compliance and vaccine coverage at the same visit. Recently, we sought to generate a vaccine vector by incorporating multiple epitopes into the rotavirus group antigenic protein, VP6. In the present study, a foreign epitope presenting a system using VP6 as a vector was created with six sites on the outer surface of the vector that could be used for insertion of foreign epitopes, and three VP6-based PV1 epitope chimeric proteins were constructed. The chimeric proteins were confirmed by immunoblot, immunofluorescence assay, and injected into guinea pigs to analyze the epitope-specific humoral response. Results showed that these chimeric proteins reacted with anti-VP6F and -PV1 antibodies, and elicited antibodies against both proteins in guinea pigs. Antibodies against the chimeric proteins carrying PV1 epitopes neutralized rotavirus Wa and PV1 infection in vitro. Our study contributes to a better understanding of the use of VP6-based vectors as multiple-epitope delivery vehicles and the epitopes displayed in this form could be considered for development of epitope-based vaccines against rotavirus and poliovirus.

VpStyA1/VpStyA2B of Variovorax paradoxus EPS: An Aryl Alkyl Sulfoxidase Rather than a Styrene Epoxidizing Monooxygenase

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Dirk Tischler

2018-04-01

Full Text Available Herein we describe the first representative of an E2-type two-component styrene monooxygenase of proteobacteria. It comprises a single epoxidase protein (VpStyA1 and a two domain protein (VpStyA2B harboring an epoxidase (A2 and a FAD-reductase (B domain. It was annotated as VpStyA1/VpStyA2B of Variovorax paradoxus EPS. VpStyA2B serves mainly as NADH:FAD-oxidoreductase. A Km of 33.6 ± 4.0 µM for FAD and a kcat of 22.3 ± 1.1 s−1 were determined and resulted in a catalytic efficiency (kcat Km−1 of 0.64 s−1 μM−1. To investigate its NADH:FAD-oxidoreductase function the linker between A2- and B-domain (AREAV was mutated. One mutant (AAAAA showed 18.7-fold higher affinity for FAD (kcat Km−1 of 5.21 s−1 μM−1 while keeping wildtype NADH-affinity and -oxidation activity. Both components, VpStyA2B and VpStyA1, showed monooxygenase activity on styrene of 0.14 U mg−1 and 0.46 U mg−1, as well as on benzyl methyl sulfide of 1.62 U mg−1 and 3.11 U mg−1, respectively. The high sulfoxidase activity was the reason to test several thioanisole-like substrates in biotransformations. VpStyA1 showed high substrate conversions (up to 95% in 2 h and produced dominantly (S-enantiomeric sulfoxides of all tested substrates. The AAAAA-mutant showed a 1.6-fold increased monooxygenase activity. In comparison, the GQWCSQY-mutant did neither show monooxygenase nor efficient FAD-reductase activity. Hence, the linker between the two domains of VpStyA2B has effects on the reductase as well as on the monooxygenase performance. Overall, this monooxygenase represents a promising candidate for biocatalyst development and studying natural fusion proteins.

Surface-displayed VP2 of IBDV on Lactobacillus casei%干酪乳杆菌表面展示IBDV VP2蛋白

Institute of Scientific and Technical Information of China (English)

林红丽; 王嵩; 王宇鹏; 栾云艳; 余丽芸; 侯喜林

2014-01-01

旨在利用pLA载体将IBDV B87株VP2蛋白展示在干酪乳杆菌表面.首先通过RT-PCR扩增VP2基因片段,测序鉴定正确后,将目的片段构建在pLA穿梭质粒上,命名为pLA-VP2,然后将重组质粒电转化到干酪乳杆菌中,构建IBDV重组干酪乳杆菌.应用SDS-PAGE检测重组菌表达目的蛋白VP2,得到约90 kDa的融合蛋白,与预期结果相符.Western blot结果显示VP2蛋白可以与抗IBDV多抗血清发生特异性反应,具有很好反应原性.流式细胞仪的散射光检测器可以捕捉到pLA-VP2重组菌菌体表面的荧光,且重组菌荧光强度强于pLA菌对照组,应用正态分布函数计算pgsA-VP2融合蛋白的表达效率,约为80％.结果表明IBDV VP2蛋白成功展示在干酪乳杆菌表面.

Human rotavirus strains bearing VP4 gene P[6] allele recovered from asymptomatic or symptomatic infections share similar, if not identical, VP4 neutralization specificities

International Nuclear Information System (INIS)

Hoshino, Yasutaka; Jones, Ronald W.; Ross, Jerri; Santos, Norma; Kapikian, Albert Z.

2003-01-01

A rotavirus VP4 gene P[6] allele has been documented in a number of countries to be characteristically associated with an endemic predominantly asymptomatic infection in neonates in maternity hospital nurseries. The mechanisms underlying the endemicity and asymptomatic nature of such neonatal infections remain unknown. Rotavirus strains sharing this same P genotype, however, have more recently been recovered from an increasing number of symptomatic diarrheal episodes in infants and young children in various parts of the world. Previously, we have shown that an asymptomatic P[6] rotavirus neonatal infection is not associated with a unique VP7 (G) serotype but may occur in conjunction with various G types. Although amino acid sequence comparisons of the VP4 gene between selected 'asymptomatic' and 'symptomatic' P[6] rotavirus strains have been reported and yielded information concerning their VP4 genotypes, serotypic comparisons of the outer capsid spike protein VP4 of such viruses have not been studied systematically by two-way cross-neutralizations. We determined the VP4 neutralization specificities of four asymptomatic and four symptomatic P[6] strains: two each of asymptomatic and symptomatic strains by two-way tests, and two each of additional asymptomatic and symptomatic strains by one-way tests. Both asymptomatic and symptomatic P[6] strains were shown to bear similar, if not identical, VP4 neutralization specificities. Thus, P[6] rotavirus strains causing asymptomatic or symptomatic infections did not appear to belong to unique P (VP4) serotypes. In addition, a close VP4 serotypic relationship between human P[6] rotavirus strains and the porcine P[6] rotavirus Gottfried strain was confirmed

Immunogenicity of virus-like particles containing modified goose parvovirus VP2 protein.

Science.gov (United States)

Chen, Zongyan; Li, Chuanfeng; Zhu, Yingqi; Wang, Binbin; Meng, Chunchun; Liu, Guangqing

2012-10-01

The major capsid protein VP2 of goose parvovirus (GPV) expressed using a baculovirus expression system (BES) assembles into virus-like particles (VLPs). To optimize VP2 gene expression in Sf9 cells, we converted wild-type VP2 (VP2) codons into codons that are more common in insect genes. This change greatly increased VP2 protein production in Sf9 cells. The protein generated from the codon-optimized VP2 (optVP2) was detected by immunoblotting and an indirect immunofluorescence assay (IFA). Transmission electron microscopy analysis revealed the formation of VLPs. These findings indicate that optVP2 yielded stable and high-quality VLPs. Immunogenicity assays revealed that the VLPs are highly immunogenic, elicit a high level of neutralizing antibodies and provide protection against lethal challenge. The antibody levels appeared to be directly related to the number of GP-Ag-positive hepatocytes. The variation trends for GP-Ag-positive hepatocytes were similar in the vaccine groups. In comparison with the control group, the optVP2 VLPs groups exhibited obviously better responses. These data indicate that the VLPs retained immunoreactivity and had strong immunogenicity in susceptible geese. Thus, GPV optVP2 appears to be a good candidate for the vaccination of goslings. Copyright © 2012 Elsevier B.V. All rights reserved.

Dynamic phosphorylation of Ebola virus VP30 in NP-induced inclusion bodies.

Science.gov (United States)

Lier, Clemens; Becker, Stephan; Biedenkopf, Nadine

2017-12-01

Zaire Ebolavirus (EBOV) causes a severe feverish disease with high case fatality rates. Transcription of EBOV is dependent on the activity of the nucleocapsid protein VP30 which represents an essential viral transcription factor. Activity of VP30 is regulated via phosphorylation at six N-terminal serine residues. Recent data demonstrated that dynamic phosphorylation and dephosphorylation of serine residue 29 is essential for transcriptional support activity of VP30. To analyze the spatio/temporal dynamics of VP30 phosphorylation, we generated a peptide antibody recognizing specifically VP30 phosphorylated at serine 29. Using this antibody we could demonstrate that (i) the majority of VP30 molecules in EBOV-infected cells is dephosphorylated at the crucial position serine 29, (ii) both, VP30 phosphorylation and dephosphorylation take place in viral inclusion bodies that are induced by the nucleoprotein NP and (iii) NP influences the phosphorylation state of VP30. Copyright © 2017 Elsevier Inc. All rights reserved.

Genetic diversity of the VP1/VP2 gene of canine parvovirus type 2b amplified from clinical specimens in Brazil Diversidade genética no gene VP1/VP2 do parvovirus canino tipo 2b amplificado de material clínico no Brasil

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Cesar A. D. Pereira

2000-10-01

Full Text Available We evaluated the genetic diversity in the VP1/VP2 gene of CPV type 2b isolates from symptomatic dogs in Brazil. A total of 21 isolates collected from 1990 through 1995 previously typed as CPV2b by PCR assay were studied. Overall we found a high degree of similarity among sequences from different CPV clinical isolates collected. Genetic analysis of this selected region gave no indication of a specific Brazilian parvovirus lineage.Neste estudo foi avaliada a diversidade genética no gene VP1/VP2 do parvovírus canino tipo 2b a partir de amostras isoladas de cães sintomáticos no Brasil. Foram estudadas 21 amostras coletadas no período de 1990 à 1995, previamente caracterizadas como CPV 2b pela técnica de PCR. Observou-se alto grau de similaridade entre as seqüências estudadas e a análise genética da região selecionada não indicou a presença de uma linhagem brasileira específica.

Intracellular trafficking of VP22 in bovine herpesvirus-1 infected cells

International Nuclear Information System (INIS)

Lobanov, Vladislav A.; Babiuk, Lorne A.; Drunen Littel-van den Hurk, Sylvia van

2010-01-01

The intracellular trafficking of different VP22-enhanced yellow fluorescent protein (EYFP) fusion proteins expressed by bovine herpesvirus-1 (BHV-1) recombinants was examined by live-cell imaging. Our results demonstrate that (i) the fusion of EYFP to the C terminus of VP22 does not alter the trafficking of the protein in infected cells, (ii) VP22 expressed during BHV-1 infection translocates to the nucleus through three different pathways, namely early mitosis-dependent nuclear translocation, late massive nuclear translocation that follows a prolonged cytoplasmic stage of the protein in non-mitotic cells, and accumulation of a small subset of VP22 in discrete dot-like nuclear domains during its early cytoplasmic stage, (iii) the addition of the SV40 large-T-antigen nuclear localization signal (NLS) to VP22-EYFP abrogates its early cytoplasmic stage, and (iv) the VP22 131 PRPR 134 NLS is not required for the late massive nuclear translocation of the protein, but this motif is essential for the targeting of VP22 to discrete dot-like nuclear domains during the early cytoplasmic stage. These results show that the amount of VP22 in the nucleus is precisely regulated at different stages of BHV-1 infection and suggest that the early pathways of VP22 nuclear accumulation may be more relevant to the infection process as the late massive nuclear influx starts when most of the viral progeny has already emerged from the cell.

Proteomic analysis reveals differential accumulation of small heat shock proteins and late embryogenesis abundant proteins between ABA-deficient mutant vp5 seeds and wild-type Vp5 seeds in maize

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Xiaolin eWu

2015-01-01

Full Text Available ABA is a major plant hormone that plays important roles during many phases of plant life cycle, including seed development, maturity and dormancy, and especially the acquisition of desiccation tolerance. Understanding of the molecular basis of ABA-mediated plant response to stress is of interest not only in basic research on plant adaptation but also in applied research on plant productivity. Maize mutant viviparous-5 (vp5, deficient in ABA biosynthesis in seeds, is a useful material for studying ABA-mediated response in maize. Due to carotenoid deficiency, vp5 endosperm is white, compared to yellow Vp5 endosperm. However, the background difference at proteome level between vp5 and Vp5 seeds is unclear. This study aimed to characterize proteome alterations of maize vp5 seeds and to identify ABA-dependent proteins during seed maturation. We compared the embryo and endosperm proteomes of vp5 and Vp5 seeds by gel-based proteomics. Up to 46 protein spots, most in embryos, were found to be differentially accumulated between vp5 and Vp5. The identified proteins included small heat shock proteins (sHSPs, late embryogenesis abundant (LEA proteins, stress proteins, storage proteins and enzymes among others. However, EMB564, the most abundant LEA protein in maize embryo, accumulated in comparable levels between vp5 and Vp5 embryos, which contrasted to previously characterized, greatly lowered expression of emb564 mRNA in vp5 embryos. Moreover, LEA proteins and sHSPs displayed differential accumulations in vp5 embryos: six out of eight identified LEA proteins decreased while nine sHSPs increased in abundance. Finally, we discussed the possible causes of global proteome alterations, especially the observed differential accumulation of identified LEA proteins and sHSPs in vp5 embryos. The data derived from this study provides new insight into ABA-dependent proteins and ABA-mediated response during maize seed maturation.

Phylogenetic similarity of the canine parvovirus wild-type isolates on the basis of VP1/VP2 gene fragment sequence analysis.

Science.gov (United States)

Rypul, K; Chmielewski, R; Smielewska-Loś, E; Klimentowski, S

2002-04-01

Biological material was taken from dogs with diarrhoea. Faecal samples were taken from within live animals and intestinal tract fragments (i.e. small intestine, and stomach) were taken from dead animals. In total, 18 specimens were investigated from dogs housed alone or in large groups. To test for the presence of the virus, latex (On Site Biotech, Uppsala, Sweden) and direct immunofluorescence tests were performed. At the same time, polymerase chain reaction (PCR) with primers complementary to a conservative region of VP1/VP2 was carried out. The products of amplification were analysed on 2% agarose gel. The purified products were cloned with the Template Generation System (Finnzymes, Espoo, Finland) using a transposition reaction and positive clones were searched using the 'colony screening by PCR' method. The sequencing gave 12 sequences of VP1/VP2 gene fragments that were of high similarity. Among the 12 analysed sequences, six exhibited 88% similarity, four exhibited 100% similarity and two exhibited 71% similarity.

Structural basis for Marburg virus VP35-mediated immune evasion mechanisms

Energy Technology Data Exchange (ETDEWEB)

Ramanan, Parameshwaran; Edwards, Megan R.; Shabman, Reed S.; Leung, Daisy W.; Endlich-Frazier, Ariel C.; Borek, Dominika M.; Otwinowski, Zbyszek; Liu, Gai; Huh, Juyoung; Basler, Christopher F.; Amarasinghe, Gaya K. [Sinai; (WU-MED); (UTSMC)

2013-07-22

Filoviruses, marburgvirus (MARV) and ebolavirus (EBOV), are causative agents of highly lethal hemorrhagic fever in humans. MARV and EBOV share a common genome organization but show important differences in replication complex formation, cell entry, host tropism, transcriptional regulation, and immune evasion. Multifunctional filoviral viral protein (VP) 35 proteins inhibit innate immune responses. Recent studies suggest double-stranded (ds)RNA sequestration is a potential mechanism that allows EBOV VP35 to antagonize retinoic-acid inducible gene-I (RIG-I) like receptors (RLRs) that are activated by viral pathogen–associated molecular patterns (PAMPs), such as double-strandedness and dsRNA blunt ends. Here, we show that MARV VP35 can inhibit IFN production at multiple steps in the signaling pathways downstream of RLRs. The crystal structure of MARV VP35 IID in complex with 18-bp dsRNA reveals that despite the similar protein fold as EBOV VP35 IID, MARV VP35 IID interacts with the dsRNA backbone and not with blunt ends. Functional studies show that MARV VP35 can inhibit dsRNA-dependent RLR activation and interferon (IFN) regulatory factor 3 (IRF3) phosphorylation by IFN kinases TRAF family member-associated NFkb activator (TANK) binding kinase-1 (TBK-1) and IFN kB kinase e (IKKe) in cell-based studies. We also show that MARV VP35 can only inhibit RIG-I and melanoma differentiation associated gene 5 (MDA5) activation by double strandedness of RNA PAMPs (coating backbone) but is unable to inhibit activation of RLRs by dsRNA blunt ends (end capping). In contrast, EBOV VP35 can inhibit activation by both PAMPs. Insights on differential PAMP recognition and inhibition of IFN induction by a similar filoviral VP35 fold, as shown here, reveal the structural and functional plasticity of a highly conserved virulence factor.

VP3 is crucial for the stability of Nora virus virions.

Science.gov (United States)

Sadanandan, Sajna Anand; Ekström, Jens-Ola; Jonna, Venkateswara Rao; Hofer, Anders; Hultmark, Dan

2016-09-02

Nora virus is an enteric virus that causes persistent, non-pathological infection in Drosophila melanogaster. It replicates in the fly gut and is transmitted via the fecal-oral route. Nora virus has a single-stranded positive-sense RNA genome, which is translated in four open reading frames. Reading frame three encodes the VP3 protein, the structure and function of which we have investigated in this work. We have shown that VP3 is a trimer that has an α-helical secondary structure, with a functionally important coiled-coil domain. In order to identify the role of VP3 in the Nora virus life cycle, we constructed VP3-mutants using the cDNA clone of the virus. Our results show that VP3 does not have a role in the actual assembly of the virus particles, but virions that lack VP3 or harbor VP3 with a disrupted coiled coil domain are incapable of transmission via the fecal-oral route. Removing the region downstream of the putative coiled coil appears to have an effect on the fitness of the virus but does not hamper its replication or transmission. We also found that the VP3 protein and particularly the coiled coil domain are crucial for the stability of Nora virus virions when exposed to heat or proteases. Hence, we propose that VP3 is imperative to Nora virus virions as it confers stability to the viral capsid. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

Hydrometer in the mantle: dln(Vs)/dln(Vp)

Science.gov (United States)

Li, L.; Weidner, D. J.

2010-12-01

The absorption of water into nominally non-hydrous phases is the probable storage mechanism of hydrogen throughout most of the mantle. Thus the water capacity in the mantle is greatest in the transition zone owing to the large water-solubility of ringwoodite and wadsleyite. However, the actual amount of water that is stored there is highly uncertain. Since water is probably brought down by subduction activity, it’s abundance is probably laterally variable. Thus, a metric that is sensitive to variations of water content are good candidates for hydrometers. Here we evaluate the parameter, dln(Vs)/dln(Vp), as such a parameter. It is useful to detect lateral variations of water if the effects of hydration on the parameter are different than those of temperature or composition. We compare the value of dln(Vs)/dln(Vp) due to the temperature with that due to the water content as a function of depth for the upper mantle. We have calculated dln(Vs)/dln(Vp) due to both water and temperature using a density functional theory approach, and available experimental data. Our results indicate that dln(Vs)/dln(Vp) due to water is distinguishable from dln(Vs)/dln(Vp) due to temperature or variations in iron content, particularly in ringwoodite. The difference increases with depth and making the lower part of the transition zone most identifiable as a water reservoir.

High Resolution Vp and Vp/Vs Local Earthquake Tomography of the Val d'Agri Region (Southern Apennines, Italy).

Science.gov (United States)

Improta, L.; Bagh, S.; De Gori, P.; Pastori, M.; Piccinini, D.; Valoroso, L.; Anselmi, M.; Buttinelli, M.; Chiarabba, C.

2015-12-01

The Val d'Agri (VA) Quaternary basin in the southern Apennines extensional belt hosts the largest oilfield in onshore Europe and normal-fault systems with high (up to M7) seismogenic potential. Frequent small-magnitude swarms related to both active crustal extension and anthropogenic activity have occurred in the region. Causal factors for induced seismicity are a water impoundment with severe seasonal oscillations and a high-rate wastewater injection well. We analyzed around 1200 earthquakes (MLENI petroleum company. We used local earthquake tomography to investigate static and transient features of the crustal velocity structure and to accurately locate earthquakes. Vp and Vp/Vs models are parameterized by a 3x3x2 km spacing and well resolved down to about 12 km depth. The complex Vp model illuminates broad antiformal structures corresponding to wide ramp-anticlines involving Mesozoic carbonates of the Apulia hydrocarbon reservoir, and NW-SE trending low Vp regions related to thrust-sheet-top clastic basins. The VA basin corresponds to shallow low-Vp region. Focal mechanisms show normal faulting kinematics with minor strike slip solutions in agreement with the local extensional regime. Earthquake locations and focal solutions depict shallow (< 5 km depth) E-dipping extensional structures beneath the artificial lake located in the southern sector of the basin, and along the western margin of the VA. A few swarms define relatively deep transfer structures accommodating the differential extension between main normal faults. The spatio-temporal distribution of around 220 events correlates with wastewater disposal activity, illuminating a NE-dipping fault between 2-5 km depth in the carbonate reservoir. The fault measures 5 km along dip and corresponds to a pre-existing thrust fault favorably oriented with respect to the local extensional field.

Estimating crustal thickness and Vp/Vs ratio with joint constraints of receiver function and gravity data

Science.gov (United States)

Shi, Lei; Guo, Lianghui; Ma, Yawei; Li, Yonghua; Wang, Weilai

2018-05-01

The technique of teleseismic receiver function H-κ stacking is popular for estimating the crustal thickness and Vp/Vs ratio. However, it has large uncertainty or ambiguity when the Moho multiples in receiver function are not easy to be identified. We present an improved technique to estimate the crustal thickness and Vp/Vs ratio by joint constraints of receiver function and gravity data. The complete Bouguer gravity anomalies, composed of the anomalies due to the relief of the Moho interface and the heterogeneous density distribution within the crust, are associated with the crustal thickness, density and Vp/Vs ratio. According to their relationship formulae presented by Lowry and Pérez-Gussinyé, we invert the complete Bouguer gravity anomalies by using a common algorithm of likelihood estimation to obtain the crustal thickness and Vp/Vs ratio, and then utilize them to constrain the receiver function H-κ stacking result. We verified the improved technique on three synthetic crustal models and evaluated the influence of selected parameters, the results of which demonstrated that the novel technique could reduce the ambiguity and enhance the accuracy of estimation. Real data test at two given stations in the NE margin of Tibetan Plateau illustrated that the improved technique provided reliable estimations of crustal thickness and Vp/Vs ratio.

A highly conserved amino acid in VP1 regulates maturation of enterovirus 71.

Directory of Open Access Journals (Sweden)

Yong-Xin Zhang

2017-09-01

Full Text Available Enterovirus 71 (EV71 is the major causative agent of hand, foot and mouth disease (HFMD in children, causing severe clinical outcomes and even death. Here, we report an important role of the highly conserved alanine residue at position 107 in the capsid protein VP1 (VP1A107 in the efficient replication of EV71. Substitutional mutations of VP1A107 significantly diminish viral growth kinetics without significant effect on viral entry, expression of viral genes and viral production. The results of mechanistic studies reveal that VP1A107 regulates the efficient cleavage of the VP0 precursor during EV71 assembly, which is required, in the next round of infection, for the transformation of the mature virion (160S into an intermediate or A-particle (135S, a key step of virus uncoating. Furthermore, the results of molecular dynamic simulations and hydrogen-bond networks analysis of VP1A107 suggest that flexibility of the VP1 BC loop or the region surrounding the VP1107 residue directly correlates with viral infectivity. It is possible that sufficient flexibility of the region surrounding the VP1107 residue favors VP0 conformational change that is required for the efficient cleavage of VP0 as well as subsequent viral uncoating and viral replication. Taken together, our data reveal the structural role of the highly conserved VP1A107 in regulating EV71 maturation. Characterization of this novel determinant of EV71 virulence would promote the study on pathogenesis of Enteroviruses.

Ebola virus VP35 blocks stress granule assembly.

Science.gov (United States)

Le Sage, Valerie; Cinti, Alessandro; McCarthy, Stephen; Amorim, Raquel; Rao, Shringar; Daino, Gian Luca; Tramontano, Enzo; Branch, Donald R; Mouland, Andrew J

2017-02-01

Stress granules (SGs) are dynamic cytoplasmic aggregates of translationally silenced mRNAs that assemble in response to environmental stress. SGs appear to play an important role in antiviral innate immunity and many viruses have evolved to block or subvert SGs components for their own benefit. Here, we demonstrate that intracellular Ebola virus (EBOV) replication and transcription-competent virus like particles (trVLP) infection does not lead to SG assembly but leads to a blockade to Arsenite-induced SG assembly. Moreover we show that EBOV VP35 represses the assembly of canonical and non-canonical SGs induced by a variety of pharmacological stresses. This SG blockade requires, at least in part, the C-terminal domain of VP35. Furthermore, results from our co-immunoprecipitation studies indicate that VP35 interacts with multiple SG components, including G3BP1, eIF3 and eEF2 through a stress- and RNA-independent mechanism. These data suggest a novel function for EBOV VP35 in the repression of SG assembly. Copyright © 2016 Elsevier Inc. All rights reserved.

Comparison of VP broadband tiltmeter and VS vertical pendulum tiltmeter

OpenAIRE

Ma, Wugang; Wu, Yanxia; Zhao, Huiqin

2015-01-01

Vertical pendulum (VP) tiltmeter is a kind of earthquake precursor observation equipment, which is used to record the interaction force associated with astronomical tidal tilts caused. Currently, VP broadband tiltmeter and vertical sensor (VS) vertical pendulum tiltmeter are primarily used. In this paper, we compare the two different instruments by using four aspects—mechanical structure, circuitry, zeroing, and bandwidth—based on their working principles and applications. We conclude that VP...

Truncated forms of viral VP2 proteins fused to EGFP assemble into fluorescent parvovirus-like particles

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Vuento Matti

2006-12-01

Full Text Available Abstract Fluorescence correlation spectroscopy (FCS monitors random movements of fluorescent molecules in solution, giving information about the number and the size of for example nano-particles. The canine parvovirus VP2 structural protein as well as N-terminal deletion mutants of VP2 (-14, -23, and -40 amino acids were fused to the C-terminus of the enhanced green fluorescent protein (EGFP. The proteins were produced in insect cells, purified, and analyzed by western blotting, confocal and electron microscopy as well as FCS. The non-truncated form, EGFP-VP2, diffused with a hydrodynamic radius of 17 nm, whereas the fluorescent mutants truncated by 14, 23 and 40 amino acids showed hydrodynamic radii of 7, 20 and 14 nm, respectively. These results show that the non-truncated EGFP-VP2 fusion protein and the EGFP-VP2 constructs truncated by 23 and by as much as 40 amino acids were able to form virus-like particles (VLPs. The fluorescent VLP, harbouring VP2 truncated by 23 amino acids, showed a somewhat larger hydrodynamic radius compared to the non-truncated EGFP-VP2. In contrast, the construct containing EGFP-VP2 truncated by 14 amino acids was not able to assemble into VLP-resembling structures. Formation of capsid structures was confirmed by confocal and electron microscopy. The number of fluorescent fusion protein molecules present within the different VLPs was determined by FCS. In conclusion, FCS provides a novel strategy to analyze virus assembly and gives valuable structural information for strategic development of parvovirus-like particles.

W 2 and Q 2 dependence of charged hadron and pion multiplicities in vp andbar vp charged current interactionscharged current interactions

Science.gov (United States)

Jones, G. T.; Jones, R. W. L.; Morrison, D. R. O.; Mobayyen, M. M.; Wainstein, S.; Aderholz, M.; Hantke, D.; Hoffmann, E.; Katz, U. F.; Kern, J.; Schmitz, N.; Wittek, W.; Allport, P.; Borner, H. P.; Myatt, G.; Radojicic, D.; Bullock, F. W.; Burke, S.

1990-03-01

Using data on vp andbar vp charged current interactions from a bubble chamber experiment with BEBC at CERN, the average multiplicities of charged hadrons and pions are determined as functions of W 2 and Q 2. The analysis is based on ˜20000 events with incident v and ˜10000 events with incidentbar v. In addition to the known dependence of the average multiplicity on W 2 a weak dependence on Q 2 for fixed intervals of W is observed. For W>2 GeV and Q 2>0.1 GeV2 the average multiplicity of charged hadrons is well described by =a 1+ a 2ln( W 2/GeV2)+ a 3ln( Q 2/GeV2) with a 1=0.465±0.053, a 2=1.211±0.021, a 3=0.103±0.014 for the vp and a 1=-0.372±0.073, a 2=1.245±0.028, a 3=0.093±0.015 for thebar vp reaction.

The K 0/π- ratio and strangeness supression in v p andbar vp charged current interactions

Science.gov (United States)

Jones, G. T.; Kennedy, B. W.; O'Neale, S. W.; Böckmann, K.; Gebel, W.; Geich-Gimbel, C.; Nellen, B.; Cooper-Sarkar, A. M.; Grant, A.; Klein, H.; Morrison, D. R. O.; Schmid, P.; Wachsmuth, H.; Chima, J. S.; Mobayyen, M. M.; Talebzadeh, M.; Villalobos-Baillie, O.; Aderholz, M.; Deck, L.; Schmitz, N.; Wernhard, K. L.; Wittek, W.; Corrigan, G.; Myatt, G.; Radojicic, D.; Saitta, B.; Wells, J.; Towers, S.; Shotton, P.

1985-03-01

Neutral kaon to negative pion production ratios from vp andbar vp charged current interactions in BEBC are presented and compared with LUND fragmentation model predictions. Good agreement is obtained with a strangeness suppression factor λ=0.203±0.014(stat)±0.010(sys). No evidence is seen for an energy dependence of λ in our kinematic region.

Effects of mutations in the VP2/VP4 cleavage site of Swine vesicular disease virus on RNA encapsidation and viral infectivity

NARCIS (Netherlands)

Rebel, J.M.J.; Leendertse, C.H.; Dekker, A.; Moormann, R.J.M.

2003-01-01

We studied VP0 cleavage of Swine vesicular disease virus (SVDV), a member of the Picornaviridae using a full-length cDNA copy of the Dutch SVDV isolate. The influences of mutations, introduced at the cleavage site of SVDV, on VP0 cleavage, RNA encapsidation and viral infection were studied. Double

Structure of Rotavirus Outer-Layer Protein VP7 Bound with a Neutralizing Fab

Energy Technology Data Exchange (ETDEWEB)

Aoki, Scott T.; Settembre, Ethan C.; Trask, Shane D.; Greenberg, Harry B.; Harrison, Stephen C.; Dormitzer, Philip R.; (Stanford-MED); (CH-Boston)

2009-06-17

Rotavirus outer-layer protein VP7 is a principal target of protective antibodies. Removal of free calcium ions (Ca{sup 2+}) dissociates VP7 trimers into monomers, releasing VP7 from the virion, and initiates penetration-inducing conformational changes in the other outer-layer protein, VP4. We report the crystal structure at 3.4 angstrom resolution of VP7 bound with the Fab fragment of a neutralizing monoclonal antibody. The Fab binds across the outer surface of the intersubunit contact, which contains two Ca{sup 2+} sites. Mutations that escape neutralization by other antibodies suggest that the same region bears the epitopes of most neutralizing antibodies. The monovalent Fab is sufficient to neutralize infectivity. We propose that neutralizing antibodies against VP7 act by stabilizing the trimer, thereby inhibiting the uncoating trigger for VP4 rearrangement. A disulfide-linked trimer is a potential subunit immunogen.

The Role of Exosomal VP40 in Ebola Virus Disease.

Science.gov (United States)

Pleet, Michelle L; DeMarino, Catherine; Lepene, Benjamin; Aman, M Javad; Kashanchi, Fatah

2017-04-01

Ebola virus (EBOV) can cause a devastating hemorrhagic disease, leading to death in a short period of time. After infection, the resulting EBOV disease results in high levels of circulating cytokines, endothelial dysfunction, coagulopathy, and bystander lymphocyte apoptosis in humans and nonhuman primates. The VP40 matrix protein of EBOV is essential for viral assembly and budding from the host cell. Recent data have shown that VP40 exists in the extracellular environment, including in exosomes, and exosomal VP40 can impact the viability of recipient immune cells, including myeloid and T cells, through the regulation of the RNAi and endosomal sorting complexes required for transport pathways. In this study, we discuss the latest findings of the impact of exosomal VP40 on immune cells in vitro and its potential implications for pathogenesis in vivo.

A Virtual Private Local PCN Ring Network Based on ATM VP Cross—Connection

Institute of Scientific and Technical Information of China (English)

LinBin; MaYingjun; 等

1995-01-01

Avirtual private local PCNring network (VPLPR)is proposed .VPLPR is a virtual logic ring seuved for digital cordless telephone system and it works on ATM VP cross-connection mechanism.Full-distributed data bases are organized for visitor location registers(VLR)and home location register(HLR).The signaling protocols are compatible upward to B-ISDN. The architecture and some of the main characteristics of VPLPR are given.How to configure the ATM VP cross-connection ring is described.And then a protocol conversion between STM frames and ATMcells in base station controller(BSC)is presented.

Immunogenicity and efficacy in mice of an adenovirus-based bicistronic rotavirus vaccine expressing NSP4 and VP7.

Science.gov (United States)

Xie, Li; Yan, Min; Wang, Xiaonan; Ye, Jing; Mi, Kai; Yan, Shanshan; Niu, Xianglian; Li, Hongjun; Sun, Maosheng

2015-12-02

NSP4 and VP7 are important functional proteins of rotavirus. Proper combination of viral gene expression is favorable to improving the protection effect of subunit vaccine. In the present study, We evaluated the immunogenicity and efficacy of the bicistronic recombinant adenovirus (rAd-NSP4-VP7) and two single-gene expressing adenoviruses (rAd-NSP4, rAd-VP7). The three adenovirus vaccines were used to immunize mice by intramuscular or intranasal administration. The data showed significant increases in serum antibodies, T lymphocyte subpopulations proliferation, and cytokine secretions of splenocyte in all immunized groups. However, the serum IgA and neutralizing antibody levels of the rAd-NSP4-VP7 or rAd-VP7 groups were significantly higher than those of the rAd-NSP4, while the splenocyte numbers of IFN-γ secretion in the rAd-NSP4-VP7 or rAd-NSP4 groups was greater than that of the rAd-VP7. Furthermore, the efficacy evaluation in a suckling mice model indicated that only rAd-NSP4-VP7 conferred significant protection against rotavirus shedding challenge. These results suggest that the co-expression of NSP4 and VP7 in an adenovirus vector induce both humoral and cell-mediated immune responses efficiently, and provide potential efficacy for protection against rotavirus disease. It is possible to represent an efficacious subunits vaccine strategy for control of rotavirus infection and transmission. Copyright © 2015 Elsevier B.V. All rights reserved.

VP Market surub tarnijale karme tingimusi / Andres Kärssin, Ave Paavo

Index Scriptorium Estoniae

Kärssin, Andres, 1971-

2004-01-01

Tänaseks Eestis 6 T-Marketi kauplust avanud Baltimaade suurim jaekaubanduskett VP Market saatis Eesti ettevõtetele ja maaletoojatele tarnelepingu vormi, mis sisaldab harjumuspärasest rohkem tingimusi ja trahve. Lisa: Detailsed nõuded ja trahvid. Vt. samas: T-Market liigutab hindu kõige lähema konkurendi järgi; VP Market on Eestis praegu kuue poega; Tõnis Arnover. VP Market harjunud jõudu kasutama

Characterization and interactome study of white spot syndrome virus envelope protein VP11.

Directory of Open Access Journals (Sweden)

Wang-Jing Liu

Full Text Available White spot syndrome virus (WSSV is a large enveloped virus. The WSSV viral particle consists of three structural layers that surround its core DNA: an outer envelope, a tegument and a nucleocapsid. Here we characterize the WSSV structural protein VP11 (WSSV394, GenBank accession number AF440570, and use an interactome approach to analyze the possible associations between this protein and an array of other WSSV and host proteins. Temporal transcription analysis showed that vp11 is an early gene. Western blot hybridization of the intact viral particles and fractionation of the viral components, and immunoelectron microscopy showed that VP11 is an envelope protein. Membrane topology software predicted VP11 to be a type of transmembrane protein with a highly hydrophobic transmembrane domain at its N-terminal. Based on an immunofluorescence assay performed on VP11-transfected Sf9 cells and a trypsin digestion analysis of the virion, we conclude that, contrary to topology software prediction, the C-terminal of this protein is in fact inside the virion. Yeast two-hybrid screening combined with co-immunoprecipitation assays found that VP11 directly interacted with at least 12 other WSSV structural proteins as well as itself. An oligomerization assay further showed that VP11 could form dimers. VP11 is also the first reported WSSV structural protein to interact with the major nucleocapsid protein VP664.

Human transbodies to VP40 inhibit cellular egress of Ebola virus-like particles

International Nuclear Information System (INIS)

Teimoori, Salma; Seesuay, Watee; Jittavisutthikul, Surasak; Chaisri, Urai; Sookrung, Nitat; Densumite, Jaslan; Saelim, Nawannaporn; Chulanetra, Monrat; Maneewatch, Santi; Chaicumpa, Wanpen

2016-01-01

A direct acting anti-Ebola agent is needed. VP40, a conserved protein across Ebolavirus (EBOV) species has several pivotal roles in the virus life cycle. Inhibition of VP40 functions would lessen the virion integrity and interfere with the viral assembly, budding, and spread. In this study, cell penetrable human scFvs (HuscFvs) that bound to EBOV VP40 were produced by phage display technology. Gene sequences coding for VP40-bound-HuscFvs were subcloned from phagemids into protein expression plasmids downstream to a gene of cell penetrating peptide, i.e., nonaarginine (R9). By electron microscopy, transbodies from three clones effectively inhibited egress of the Ebola virus-like particles from human hepatic cells transduced with pseudo-typed-Lentivirus particles carrying EBOV VP40 and GP genes. Computerized simulation indicated that the effective HuscFvs bound to multiple basic residues in the cationic patch of VP40 C-terminal domain which are important in membrane-binding for viral matrix assembly and virus budding. The transbodies bound also to VP40 N-terminal domain and L domain peptide encompassed the PTAPPEY (WW binding) motif, suggesting that they might confer VP40 function inhibition through additional mechanism(s). The generated transbodies are worthwhile tested with authentic EBOV before developing to direct acting anti-Ebola agent for preclinical and clinical trials. - Highlights: • Cell penetrable human scFvs (transbodies) to Ebolavirus (EBOV) VP40 were produced. • The transbodies inhibited egress of EBOV-like particles (VLPs) from human hepatocytes. • They interacted with VP40 CTD basic residues important for plasma membrane binding. • And hence interfere with viral matrix assembly and viral progeny budding. • This is the first report on human antibodies that target intracellular EBOV VP40.

Structural and functional characterization of Reston Ebola virus VP35 interferon inhibitory domain.

Science.gov (United States)

Leung, Daisy W; Shabman, Reed S; Farahbakhsh, Mina; Prins, Kathleen C; Borek, Dominika M; Wang, Tianjiao; Mühlberger, Elke; Basler, Christopher F; Amarasinghe, Gaya K

2010-06-11

Ebolaviruses are causative agents of lethal hemorrhagic fever in humans and nonhuman primates. Among the filoviruses characterized thus far, Reston Ebola virus (REBOV) is the only Ebola virus that is nonpathogenic to humans despite the fact that REBOV can cause lethal disease in nonhuman primates. Previous studies also suggest that REBOV is less effective at inhibiting host innate immune responses than Zaire Ebola virus (ZEBOV) or Marburg virus. Virally encoded VP35 protein is critical for immune suppression, but an understanding of the relative contributions of VP35 proteins from REBOV and other filoviruses is currently lacking. In order to address this question, we characterized the REBOV VP35 interferon inhibitory domain (IID) using structural, biochemical, and virological studies. These studies reveal differences in double-stranded RNA binding and interferon inhibition between the two species. These observed differences are likely due to increased stability and loss of flexibility in REBOV VP35 IID, as demonstrated by thermal shift stability assays. Consistent with this finding, the 1.71-A crystal structure of REBOV VP35 IID reveals that it is highly similar to that of ZEBOV VP35 IID, with an overall backbone r.m.s.d. of 0.64 A, but contains an additional helical element at the linker between the two subdomains of VP35 IID. Mutations near the linker, including swapping sequences between REBOV and ZEBOV, reveal that the linker sequence has limited tolerance for variability. Together with the previously solved ligand-free and double-stranded-RNA-bound forms of ZEBOV VP35 IID structures, our current studies on REBOV VP35 IID reinforce the importance of VP35 in immune suppression. Functional differences observed between REBOV and ZEBOV VP35 proteins may contribute to observed differences in pathogenicity, but these are unlikely to be the major determinant. However, the high level of similarity in structure and the low tolerance for sequence variability, coupled

Leedu hiiglane VP Market läheb börsile / Mihkel Salu

Index Scriptorium Estoniae

Salu, Mihkel

2005-01-01

Intervjuust VR Marketi uue juhi Gintaras Marcinkieviciusega. VP-Marketi emafirma Vilniaus Prekyba ootab Eesti turul järgmisel aastal läbimurret, Eestis kavatsetakse avada esimesed super- ja hüpermarketid, VP Market ja samasse kontserni kuuluv apteegikett Euroapteek lähevad börsile. Lisa: VP Market kasvab kiirelt

Down-Regulation of Na+/K+ ATPase Activity by Human Parvovirus B19 Capsid Protein VP1

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Ahmad Almilaji

2013-05-01

Full Text Available Background/Aims: Human parvovirus B19 (B19V may cause inflammatory cardiomyopathy (iCMP which is accompanied by endothelial dysfunction. The B19V capsid protein VP1 contains a lysophosphatidylcholine producing phospholipase A2 (PLA sequence. Lysophosphatidylcholine has in turn been shown to inhibit Na+/K+ ATPase. The present study explored whether VP1 modifies Na+/K+ ATPase activity. Methods: Xenopus oocytes were injected with cRNA encoding VP1 isolated from a patient suffering from fatal B19V-iCMP or cRNA encoding PLA2-negative VP1 mutant (H153A and K+ induced pump current (Ipump as well as ouabain-inhibited current (Iouabain both reflecting Na+/K+-ATPase activity were determined by dual electrode voltage clamp. Results: Injection of cRNA encoding VP1, but not of VP1(H153A or water, was followed by a significant decrease of both, Ipump and Iouabain in Xenopus oocytes. The effect was not modified by inhibition of transcription with actinomycin (10 µM for 36 hours but was abrogated in the presence of PLA2 specific blocker 4-bromophenacylbromide (50 µM and was mimicked by lysophosphatidylcholine (0.5 - 1 µg/ml. According to whole cell patch clamp, lysophosphatidylcholine (1 µg /ml similarly decreased Ipump in human microvascular endothelial cells (HMEC. Conclusion: The B19V capsid protein VP1 is a powerful inhibitor of host cell Na+/K+ ATPase, an effect at least partially due to phospholipase A2 (PLA2 dependent formation of lysophosphatidylcholine.

Vp-Vs relationship and amplitude variation with offset modelling of glauconitic greensand

DEFF Research Database (Denmark)

Hossain, Zakir; Mukerji, Tapan; Fabricius, Ida Lykke

2012-01-01

The relationship between Vp and Vs may be used to predict Vs where only Vp is known. Vp/Vs is also used to identify pore fluids from seismic data and amplitude variation with offset analysis. Theoretical, physical, as well as statistical empirical Vp-Vs relationships have been proposed...... modelling works well for greensand shear-wave velocity prediction. We model the seismic response of glauconitic greensand by using laboratory data from the Nini field. Our studies here reveal that brine-saturated glauconitic greensand can have a similar seismic response to that from oil-saturated quartzitic...

The Ebola Virus VP30-NP Interaction Is a Regulator of Viral RNA Synthesis

Energy Technology Data Exchange (ETDEWEB)

Kirchdoerfer, Robert N.; Moyer, Crystal L.; Abelson, Dafna M.; Saphire, Erica Ollmann (Scripps)

2016-10-18

Filoviruses are capable of causing deadly hemorrhagic fevers. All nonsegmented negative-sense RNA-virus nucleocapsids are composed of a nucleoprotein (NP), a phosphoprotein (VP35) and a polymerase (L). However, the VP30 RNA-synthesis co-factor is unique to the filoviruses. The assembly, structure, and function of the filovirus RNA replication complex remain unclear. Here, we have characterized the interactions of Ebola, Sudan and Marburg virus VP30 with NP using in vitro biochemistry, structural biology and cell-based mini-replicon assays. We have found that the VP30 C-terminal domain interacts with a short peptide in the C-terminal region of NP. Further, we have solved crystal structures of the VP30-NP complex for both Ebola and Marburg viruses. These structures reveal that a conserved, proline-rich NP peptide binds a shallow hydrophobic cleft on the VP30 C-terminal domain. Structure-guided Ebola virus VP30 mutants have altered affinities for the NP peptide. Correlation of these VP30-NP affinities with the activity for each of these mutants in a cell-based mini-replicon assay suggests that the VP30-NP interaction plays both essential and inhibitory roles in Ebola virus RNA synthesis.

Comparison of VP broadband tiltmeter and VS vertical pendulum tiltmeter

Directory of Open Access Journals (Sweden)

Wugang Ma

2015-05-01

Full Text Available Vertical pendulum (VP tiltmeter is a kind of earthquake precursor observation equipment, which is used to record the interaction force associated with astronomical tidal tilts caused. Currently, VP broadband tiltmeter and vertical sensor (VS vertical pendulum tiltmeter are primarily used. In this paper, we compare the two different instruments by using four aspects—mechanical structure, circuitry, zeroing, and bandwidth—based on their working principles and applications. We conclude that VP broadband tiltmeter is more superior compared with VS vertical pendulum tiltmeter because of its higher bandwidth and degree of automation.

VP22 herpes simplex virus protein can transduce proteins into stem cells

Energy Technology Data Exchange (ETDEWEB)

Gabanyi, I.; Lojudice, F.H.; Kossugue, P.M. [Centro de Terapia Celular e Molecular, Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo, SP (Brazil); Rebelato, E. [Departamento de Fisiologia e Biofísica, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, SP (Brazil); Demasi, M.A.; Sogayar, M.C. [Centro de Terapia Celular e Molecular, Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo, SP (Brazil)

2013-02-01

The type I herpes simplex virus VP22 tegument protein is abundant and well known for its ability to translocate proteins from one cell to the other. In spite of some reports questioning its ability to translocate proteins by attributing the results observed to fixation artifacts or simple attachment to the cell membrane, VP22 has been used to deliver several proteins into different cell types, triggering the expected cell response. However, the question of the ability of VP22 to enter stem cells has not been addressed. We investigated whether VP22 could be used as a tool to be applied in stem cell research and differentiation due to its capacity to internalize other proteins without altering the cell genome. We generated a VP22.eGFP construct to evaluate whether VP22 could be internalized and carry another protein with it into two different types of stem cells, namely adult human dental pulp stem cells and mouse embryonic stem cells. We generated a VP22.eGFP fusion protein and demonstrated that, in fact, it enters stem cells. Therefore, this system may be used as a tool to deliver various proteins into stem cells, allowing stem cell research, differentiation and the generation of induced pluripotent stem cells in the absence of genome alterations.

VP22 herpes simplex virus protein can transduce proteins into stem cells

International Nuclear Information System (INIS)

Gabanyi, I.; Lojudice, F.H.; Kossugue, P.M.; Rebelato, E.; Demasi, M.A.; Sogayar, M.C.

2013-01-01

The type I herpes simplex virus VP22 tegument protein is abundant and well known for its ability to translocate proteins from one cell to the other. In spite of some reports questioning its ability to translocate proteins by attributing the results observed to fixation artifacts or simple attachment to the cell membrane, VP22 has been used to deliver several proteins into different cell types, triggering the expected cell response. However, the question of the ability of VP22 to enter stem cells has not been addressed. We investigated whether VP22 could be used as a tool to be applied in stem cell research and differentiation due to its capacity to internalize other proteins without altering the cell genome. We generated a VP22.eGFP construct to evaluate whether VP22 could be internalized and carry another protein with it into two different types of stem cells, namely adult human dental pulp stem cells and mouse embryonic stem cells. We generated a VP22.eGFP fusion protein and demonstrated that, in fact, it enters stem cells. Therefore, this system may be used as a tool to deliver various proteins into stem cells, allowing stem cell research, differentiation and the generation of induced pluripotent stem cells in the absence of genome alterations

Expression of goose parvovirus whole VP3 protein and its epitopes in Escherichia coli cells.

Science.gov (United States)

Tarasiuk, K; Woźniakowski, G; Holec-Gąsior, L

2015-01-01

The aim of this study was the expression of goose parvovirus capsid protein (VP3) and its epitopes in Escherichia coli cells. Expression of the whole VP3 protein provided an insufficient amount of protein. In contrast, the expression of two VP3 epitopes (VP3ep4, VP3ep6) in E. coli, resulted in very high expression levels. This may suggest that smaller parts of the GPV antigenic determinants are more efficiently expressed than the complete VP3 gene.

[Expression and activity determination of recombinant capsid protein VP2 gene of enterovirus type 71].

Science.gov (United States)

Huang, Xueyong; Liu, Guohua; Hu, Xiaoning; Du, Yanhua; Li, Xingle; Xu, Yuling; Chen, Haomin; Xu, Bianli

2014-04-01

To clone and express the recombinant capsid protein VP2 of enterovirus type 71 (EV71) and to identify the immune activity of expressed protein in order to build a basis for the investigation work of vaccine and diagnostic antigen. VP2 gene of EV71 was amplified by PCR, and then was cut by restriction enzyme and inserted into expression vector pMAL-c2X. The positive recombinants were transferred into E.coli TB1, the genetically engineered bacteria including pMAL-c2X-VP2 plasmids were induced by isopropyl thiogalactoside ( IPTG) , and the expression products were analyzed by SDS-PAGE and western blotting method. EV71 IgM antibody detection method by ELISA was set up, and the sensitivity and specificity of this method was assessed; 60 neutralizing antibody positive serum samples from hand foot and mouth disease (HFMD) patients were determined, of which 52 samples were positive and 8 samples were negative; a total of 88 acute phase serum samples of HFMD patients diagnosed in clinical were also detected. VP2 gene of 762 bp was obtained by PCR, the gene segment inserted into the recombinant vector was identified using restriction enzyme digestion. The recombinant vector could express a specific about 71 500 fusion protein in E.coli by SDS-PAGE. The purified recombinant protein of EV71-VP2 can react with the serum of HFMD patients to produce a specific band by western blotting. The sensitivity and specificity of ELISA was 87% and 83%, respectively. Of the 88 acute phase serum samples from children with HFMD, 48 samples (55%) were positive by the ELISA assay. VP2 gene of EV71 has been cloned and a prokaryotic high expression system for VP2 gene was successfully constructed in the present study. The recombination EV71-VP2 has well antigenicity, which could be useful for developing diagnose reagent or vaccine of EV71.

Vaccination with multimeric recombinant VP28 induces high protection against white spot syndrome virus in shrimp.

Science.gov (United States)

Taengchaiyaphum, Suparat; Nakayama, Hideki; Srisala, Jiraporn; Khiev, Ratny; Aldama-Cano, Diva January; Thitamadee, Siripong; Sritunyalucksana, Kallaya

2017-11-01

To improve the efficacy of WSSV protection, multimeric (tetrameric) recombinant VP28 (4XrVP28) was produced and tested in comparison with those of monomeric VP28 (1XrVP28). In vitro binding of either 1XrVP28 or 4XrVP28 to shrimp hemocyte surface was evident as early as 10 min after protein inoculation. Similar results were obtained in vivo when shrimp were injected with recombinant proteins that the proteins bound to the hemocyte surface could be detected since 5 min after injection. Comparison of the WSSV protection efficiencies of 1XrVP28 or 4XrVP28 were performed by injection the purified 1XrVP28 or 4XrVP28 (22.5 μg/shrimp) and WSSV inoculum (1000 copies/shrimp) into shrimp. At 10 dpi, while shrimp injected with WSSV inoculum reached 100% mortality, shrimp injected with 1XrVP28 + WSSV or 4XrVP28 + WSSV showed relative percent survival (RPS) of 67% and 81%, respectively. PCR quantification revealed high number of WSSV in the moribund shrimp of WSSV- and 1XrVP28+WSSV-injected group. In contrast, lower number of WSSV copies were found in the survivors both from 1XrVP28+WSSV- or 4XrVP28+WSSV- injected groups. Histopathological analysis demonstrated the WSSV infected lesions found in the moribund from WSSV-infected group and 1XrVP28+WSSV-injected group, but less or none in the survivors. ELISA demonstrated that 4XrVP28 exhibited higher affinity binding to rPmRab7, a WSSV binding protein essential for WSSV entry to the cell than 1XrVP28. Taken together, the protection against WSSV in shrimp could be improved by application of multimeric rVP28. Copyright © 2017 Elsevier Ltd. All rights reserved.

The role of RAD51 in etoposide (VP16) resistance in small cell lung cancer

DEFF Research Database (Denmark)

Hansen, Lasse Tengbjerg; Lundin, Cecilia; Spang-Thomsen, Mogens

2003-01-01

Etoposide (VP16) is a potent inducer of DNA double-strand breaks (DSBs) and is efficiently used in small cell lung cancer (SCLC) therapy. However, acquired VP16 resistance remains an important barrier to effective treatment. To understand the underlying mechanisms for VP16 resistance in SCLC, we...... investigated DSB repair and cellular VP16 sensitivity of SCLC cells. VP16 sensitivity and RAD51, DNA-PK(cs), topoisomerase IIalpha and P-glycoprotein protein levels were determined in 17 SCLC cell lines. In order to unravel the role of RAD51 in VP16 resistance, we cloned the human RAD51 gene, transfected SCLC...... cells with RAD51 sense or antisense constructs and measured the VP16 resistance. Finally, we measured VP16-induced DSBs in the 17 SCLC cell lines. Two cell lines exhibited a multidrug-resistant phenotype. In the other SCLC cell lines, the cellular VP16 resistance was positively correlated with the RAD51...

The Ebola Virus VP30-NP Interaction Is a Regulator of Viral RNA Synthesis.

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Robert N Kirchdoerfer

2016-10-01

Full Text Available Filoviruses are capable of causing deadly hemorrhagic fevers. All nonsegmented negative-sense RNA-virus nucleocapsids are composed of a nucleoprotein (NP, a phosphoprotein (VP35 and a polymerase (L. However, the VP30 RNA-synthesis co-factor is unique to the filoviruses. The assembly, structure, and function of the filovirus RNA replication complex remain unclear. Here, we have characterized the interactions of Ebola, Sudan and Marburg virus VP30 with NP using in vitro biochemistry, structural biology and cell-based mini-replicon assays. We have found that the VP30 C-terminal domain interacts with a short peptide in the C-terminal region of NP. Further, we have solved crystal structures of the VP30-NP complex for both Ebola and Marburg viruses. These structures reveal that a conserved, proline-rich NP peptide binds a shallow hydrophobic cleft on the VP30 C-terminal domain. Structure-guided Ebola virus VP30 mutants have altered affinities for the NP peptide. Correlation of these VP30-NP affinities with the activity for each of these mutants in a cell-based mini-replicon assay suggests that the VP30-NP interaction plays both essential and inhibitory roles in Ebola virus RNA synthesis.

Membrane association and localization dynamics of the Ebola virus matrix protein VP40.

Science.gov (United States)

Gc, Jeevan B; Gerstman, Bernard S; Chapagain, Prem P

2017-10-01

The Ebola virus matrix protein VP40 is a major structural protein that provides the scaffolding for new Ebola virus particles. For this, VP40 is first trafficked to the lower leaflet of the plasma membrane (PM) in its dimeric form. Once associated with the PM, the VP40 dimers undergo structural rearrangements and oligomerize into hexamers and filaments that make up the virus matrix. Therefore, association of the VP40 dimers and their stabilization at the PM is a crucial step in the Ebola life-cycle. To understand the molecular details of the VP40 dimer-PM interactions, we investigated the dimer association with the inner leaflet of the PM using detailed all-atom molecular dynamics (MD) simulations. The formation of the dimer-PM complex is facilitated by the interactions of the VP40 lysine residues and the anionic lipids POPS, POPI, and PIP 2 in the PM. In contrast, the dimer fails to associate with a membrane without POPS, POPI, or PIP 2 lipids. We explored the mechanisms of the association and identified important residues and lipids involved in localization and stabilization of VP40 dimers at the PM. MD simulations elucidate the role of a C-terminal α-helix alignment parallel to the lipid bilayer surface as well as the creation of membrane defects that allow partial insertion of the hydrophobic residue V276 into the membrane to further stabilize the VP40 dimer-PM complex. Understanding the mechanisms of the VP40 dimer-PM association that facilitate oligomerization can be important for potentially targeting the VP40 for small molecules that can interfere with the virus life-cycle. Copyright © 2017 Elsevier B.V. All rights reserved.

Ebola VP40 in Exosomes Can Cause Immune Cell Dysfunction.

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Pleet, Michelle L; Mathiesen, Allison; DeMarino, Catherine; Akpamagbo, Yao A; Barclay, Robert A; Schwab, Angela; Iordanskiy, Sergey; Sampey, Gavin C; Lepene, Benjamin; Nekhai, Sergei; Aman, M J; Kashanchi, Fatah

2016-01-01

Ebola virus (EBOV) is an enveloped, ssRNA virus from the family Filoviridae capable of causing severe hemorrhagic fever with up to 80-90% mortality rates. The most recent outbreak of EBOV in West Africa starting in 2014 resulted in over 11,300 deaths; however, long-lasting persistence and recurrence in survivors has been documented, potentially leading to further transmission of the virus. We have previously shown that exosomes from cells infected with HIV-1, HTLV-1 and Rift Valley Fever virus are able to transfer viral proteins and non-coding RNAs to naïve recipient cells, resulting in an altered cellular activity. In the current manuscript, we examined the effect of Ebola structural proteins VP40, GP, NP and VLPs on recipient immune cells, as well as the effect of exosomes containing these proteins on naïve immune cells. We found that VP40-transfected cells packaged VP40 into exosomes, and that these exosomes were capable of inducing apoptosis in recipient immune cells. Additionally, we show that presence of VP40 within parental cells or in exosomes delivered to naïve cells could result in the regulation of RNAi machinery including Dicer, Drosha, and Ago 1, which may play a role in the induction of cell death in recipient immune cells. Exosome biogenesis was regulated by VP40 in transfected cells by increasing levels of ESCRT-II proteins EAP20 and EAP45, and exosomal marker proteins CD63 and Alix. VP40 was phosphorylated by Cdk2/Cyclin complexes at Serine 233 which could be reversed with r-Roscovitine treatment. The level of VP40-containing exosomes could also be regulated by treated cells with FDA-approved Oxytetracycline. Additionally, we utilized novel nanoparticles to safely capture VP40 and other viral proteins from Ebola VLPs spiked into human samples using SDS/reducing agents, thus minimizing the need for BSL-4 conditions for most downstream assays. Collectively, our data indicates that VP40 packaged into exosomes may be responsible for the deregulation

Ebola VP40 in Exosomes Can Cause Immune Cell Dysfunction

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Michelle L. Pleet

2016-11-01

Full Text Available Ebola virus (EBOV is an enveloped, ssRNA virus from the family Filoviridae capable of causing severe hemorrhagic fever with up to 80–90% mortality rates. The most recent outbreak of EBOV in West Africa starting in 2014 resulted in over 11,300 deaths; however, long-lasting persistence and recurrence in survivors has been documented, potentially leading to further transmission of the virus. We have previously shown that exosomes from cells infected with HIV-1, HTLV-1 and Rift Valley Fever virus are able to transfer viral proteins and non-coding RNAs to naïve recipient cells, resulting in an altered cellular activity. In the current manuscript, we examined the effect of Ebola structural proteins VP40, GP, NP and VLPs on recipient immune cells, as well as the effect of exosomes containing these proteins on naïve immune cells. We found that VP40-transfected cells packaged VP40 into exosomes, and that these exosomes were capable of inducing apoptosis in recipient immune cells. Additionally, we show that presence of VP40 within parental cells or in exosomes delivered to naïve cells could result in the regulation of RNAi machinery including Dicer, Drosha, and Ago 1, which may play a role in the induction of cell death in recipient immune cells. Exosome biogenesis was regulated by VP40 in transfected cells by increasing levels of ESCRT-II proteins EAP20 and EAP45, and exosomal marker proteins CD63 and Alix. VP40 was phosphorylated by Cdk2/Cyclin complexes at Serine 233 which could be reversed with r-Roscovitine treatment. The level of VP40-containing exosomes could also be regulated by treated cells with FDA-approved Oxytetracycline. Additionally, we utilized novel nanoparticles to safely capture VP40 and other viral proteins from Ebola VLPs spiked into human samples using SDS/reducing agents, thus minimizing the need for BSL-4 conditions for most downstream assays. Collectively, our data indicates that VP40 packaged into exosomes may be responsible

Membrane Binding and Bending in Ebola VP40 Assembly and Egress

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Robert V Stahelin

2014-06-01

Full Text Available Lipid-enveloped viruses contain a lipid bilayer coat that protects their genome and helps to facilitate entry into the host cell. Filoviruses are lipid-enveloped viruses that have up to 90% clinical fatality and include Marbug (MARV and Ebola (EBOV. These pleomorphic filamentous viruses enter the host cell through their membrane embedded glycoprotein and then replicate using just seven genes encoded in their negative sense RNA genome. EBOV budding occurs from the inner leaflet of the plasma membrane and is driven by the matrix protein VP40, which is the most abundantly expressed protein of the virus. VP40 expressed in mammalian cells alone can trigger budding of filamentous virus-like particles (VLPs that are nearly indistinguishable from authentic EBOV. VP40, like matrix proteins from other viruses, has been shown to bind anionic lipid membranes. However, how VP40 selectively interacts with the inner leaflet of the plasma membrane and assembles into a filamentous lipid enveloped particle is mostly unknown. This article describes what is known regarding VP40 membrane interactions and what answers will fill the gaps.

Characterization and protective efficacy in an animal model of a novel truncated rotavirus VP8 subunit parenteral vaccine candidate.

Science.gov (United States)

Xue, Miaoge; Yu, Linqi; Che, Yaojian; Lin, Haijun; Zeng, Yuanjun; Fang, Mujin; Li, Tingdong; Ge, Shengxiang; Xia, Ningshao

2015-05-21

The cell-attachment protein VP8* of rotavirus is a potential candidate parenteral vaccine. However, the yield of full-length VP8 protein (VP8*, residues 1-231) expressed in Escherichia coli was low, and a truncated VP8 protein (ΔVP8*, residues 65-231) cannot elicit efficient protective immunity in a mouse model. In this study, tow novel truncated VP8 proteins, VP8-1 (residues 26-231) and VP8-2 (residues 51-231), were expressed in E. coli and evaluated for immunogenicity and protective efficacy, compared with VP8* and ΔVP8*. As well as ΔVP8*, the protein VP8-1 and VP8-2 were successfully expressed in high yield and purified in homogeneous dimeric forms, while the protein VP8* was expressed with lower yield and prone to aggregation and degradation in solution. Although the immunogenicity of the protein VP8*, VP8-1, VP8-2 and ΔVP8* was comparable, immunization of VP8* and VP8-1 elicited significantly higher neutralizing antibody titers than that of VP8-2 and ΔVP8* in mice. Furthermore, when assessed using a mouse maternal antibody model, the efficacy of VP8-1 to protect against rotavirus-induced diarrhea in pups was comparable to that of VP8*, both were dramatically higher than that of VP8-2 and ΔVP8*. Taken together, the novel truncated protein VP8-1, with increased yield, improved homogeneity and high protective efficacy, is a viable candidate for further development of a parenterally administrated prophylactic vaccine against rotavirus infection. Copyright © 2015 Elsevier Ltd. All rights reserved.

A Loop Region in the N-Terminal Domain of Ebola Virus VP40 Is Important in Viral Assembly, Budding, and Egress

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Emmanuel Adu-Gyamfi

2014-10-01

Full Text Available Ebola virus (EBOV causes viral hemorrhagic fever in humans and can have clinical fatality rates of ~60%. The EBOV genome consists of negative sense RNA that encodes seven proteins including viral protein 40 (VP40. VP40 is the major Ebola virus matrix protein and regulates assembly and egress of infectious Ebola virus particles. It is well established that VP40 assembles on the inner leaflet of the plasma membrane of human cells to regulate viral budding where VP40 can produce virus like particles (VLPs without other Ebola virus proteins present. The mechanistic details, however, of VP40 lipid-interactions and protein-protein interactions that are important for viral release remain to be elucidated. Here, we mutated a loop region in the N-terminal domain of VP40 (Lys127, Thr129, and Asn130 and find that mutations (K127A, T129A, and N130A in this loop region reduce plasma membrane localization of VP40. Additionally, using total internal reflection fluorescence microscopy and number and brightness analysis we demonstrate these mutations greatly reduce VP40 oligomerization. Lastly, VLP assays demonstrate these mutations significantly reduce VLP release from cells. Taken together, these studies identify an important loop region in VP40 that may be essential to viral egress.

A novel type of VP4 carried by a porcine rotavirus strain

International Nuclear Information System (INIS)

Liprandi, Ferdinando; Gerder, Marlene; Bastidas, Zoleida; Lopez, Jose A.; Pujol, Flor H.; Ludert, Juan E.; Joelsson, Daniel B.; Ciarlet, Max

2003-01-01

The gene encoding the VP8* trypsin-cleavage product of the VP4 protein of porcine rotavirus strain A34 was sequenced, and the predicted amino acid (aa) sequence was compared to the homologous region of all known P genotypes. The aa sequence of the VP8* of strain A34 shared low identity, ranging from 39% (bovine strain B223, P8[11]) to 76% (human strain 69M, P4[10]), with the homologous sequences of representative strains of the remaining 21 P genotypes. Phylogenetic relationships showed that the VP8* of strain A34 shares a common evolutionary lineage with those of human 69M (P4[10]) and equine H-2 (P4[12]) strains. Hyperimmune sera raised to strain A34 and to a genetic reassortant strain containing the VP4 gene from strain A34, both with high homologous neutralization titer via VP4, failed to neutralize strains representative of 15 different P genotypes. These results indicate that strain A34 should be considered as prototype of a new P genotype and serotype (P14[23]) and provide further evidence for the vast genetic and antigenic diversity of group A rotaviruses

Phylogenetic analysis of G1P[8] and G12P[8] rotavirus A samples obtained in the pre- and post-vaccine periods, and molecular modeling of VP4 and VP7 proteins.

Science.gov (United States)

Almeida, Tâmera Nunes Vieira; de Sousa, Teresinha Teixeira; da Silva, Roosevelt Alves; Fiaccadori, Fabíola Souza; Souza, Menira; Badr, Kareem Rady; de Paula Cardoso, Divina das Dôres

2017-09-01

Reduction in morbimortality rates for acute gastroenteritis (AGE) by Rotavirus A (RVA) has been observed after the introduction of vaccines, however the agent continues to circulate. The present study described the genomic characterization of the 11 dsRNA segments of two RVA samples G1P[8] obtained in the pre- and post-vaccination periods and one of G12P[8] sample (post-vaccine), compared to Rotarix™ vaccine. Analysis by molecular sequencing of the samples showed that the three samples belonged to genogroup I. In addition, the analysis of VP7 gene revealed that the samples G1 (pre-vaccine), G1 (post-vaccine) and G12 were characterized as lineages II, I and III, respectively. Regarding to VP4 and NSP4 gene it was observed that all samples belonged to lineage III, whereas for VP6 gene, the sample of the pre- and post-vaccine belonged to the lineage IV and I, respectively. Considering the VP7 gene, it was observed high nucleotide and amino acid identity for the two G1 samples when compared to Rotarix™ vaccine and lesser identity for the G12 sample. In relation to antigenic epitope of VP7 greater modifications were observed for the G12 sample in the 7-2 epitope that was confirmed by molecular modeling. On the other hand, for VP4, some changes in the 8-1 and 8-3 antigenic epitopes was observed for the three samples. This data could be interpreted as a low selective pressure exerted by vaccination in relation to G1P[8] samples and lesser protection in relation to G12P[8]. Thus, the continuous monitoring of RVA circulating samples remains important. Copyright © 2017 Elsevier B.V. All rights reserved.

Dimerization Controls Marburg Virus VP24-dependent Modulation of Host Antioxidative Stress Responses

Energy Technology Data Exchange (ETDEWEB)

Johnson, Britney; Li, Jing; Adhikari, Jagat; Edwards, Megan R.; Zhang, Hao; Schwarz, Toni; Leung, Daisy W.; Basler, Christopher F.; Gross, Michael L.; Amarasinghe, Gaya K.

2016-08-04

Marburg virus (MARV), a member of the Filoviridae family that also includes Ebola virus (EBOV), causes lethal hemorrhagic fever with case fatality rates that have exceeded 50% in some outbreaks. Within an infected cell, there are numerous host-viral interactions that contribute to the outcome of infection. Recent studies identified MARV protein 24 (mVP24) as a modulator of the host antioxidative responses, but the molecular mechanism remains unclear. Using a combination of biochemical and mass spectrometry studies, we show that mVP24 is a dimer in solution that directly binds to the Kelch domain of Kelch-like ECH-associated protein 1 (Keap1) to regulate nuclear factor (erythroid-derived 2)-like 2 (Nrf2). This interaction between Keap1 and mVP24 occurs through the Kelch interaction loop (K-Loop) of mVP24 leading to upregulation of antioxidant response element transcription, which is distinct from other Kelch binders that regulate Nrf2 activity. N-terminal truncations disrupt mVP24 dimerization, allowing monomeric mVP24 to bind Kelch with higher affinity and stimulate higher antioxidative stress response element (ARE) reporter activity. Mass spectrometry-based mapping of the interface revealed overlapping binding sites on Kelch for mVP24 and the Nrf2 proteins. Substitution of conserved cysteines, C209 and C210, to alanine in the mVP24 K-Loop abrogates Kelch binding and ARE activation. Our studies identify a shift in the monomer-dimer equilibrium of MARV VP24, driven by its interaction with Keap1 Kelch domain, as a critical determinant that modulates host responses to pathogenic Marburg viral infections.

Recombinant VP1, an Akt inhibitor, suppresses progression of hepatocellular carcinoma by inducing apoptosis and modulation of CCL2 production.

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Tai-An Chen

Full Text Available BACKGROUND: The application of viral elements in tumor therapy is one facet of cancer research. Recombinant capsid protein VP1 (rVP1 of foot-and-mouth disease virus has previously been demonstrated to induce apoptosis in cancer cell lines. Here, we aim to further investigate its apoptotic mechanism and possible anti-metastatic effect in murine models of hepatocellular carcinoma (HCC, one of the most common human cancers worldwide. METHODOLOGY/PRINCIPAL FINDINGS: Treatment with rVP1 inhibited cell proliferation in two murine HCC cell lines, BNL and Hepa1-6, with IC₅₀ values in the range of 0.1-0.2 µM. rVP1 also induced apoptosis in these cells, which was mediated by Akt deactivation and dissociation of Ku70-Bax, and resulted in conformational changes and mitochondrial translocation of Bax, leading to the activation of caspases-9, -3 and -7. Treatment with 0.025 µM rVP1, which did not affect the viability of normal hepatocytes, suppressed cell migration and invasion via attenuating CCL2 production. The production of CCL2 was modulated by Akt-dependent NF-κB activation that was decreased after rVP1 treatment. The in vivo antitumor effects of rVP1 were assessed in both subcutaneous and orthotopic mouse models of HCC in immune-competent BALB/c mice. Intratumoral delivery of rVP1 inhibited subcutaneous tumor growth as a result of increased apoptosis. Intravenous administration of rVP1 in an orthotopic HCC model suppressed tumor growth, inhibited intra-hepatic metastasis, and prolonged survival. Furthermore, a decrease in the serum level of CCL2 was observed in rVP1-treated mice. CONCLUSIONS/SIGNIFICANCE: The data presented herein suggest that, via inhibiting Akt phosphorylation, rVP1 suppresses the growth, migration, and invasion of murine HCC cells by inducing apoptosis and attenuating CCL2 production both in vitro and in vivo. Recombinant protein VP1 thus has the potential to be developed as a new therapeutic agent for HCC.

Efficacy of double-stranded RNA against white spot syndrome virus (WSSV non-structural (orf89, wsv191 and structural (vp28, vp26 genes in the Pacific white shrimp Litopenaeus vannamei

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César M. Escobedo-Bonilla

2015-04-01

Full Text Available White spot syndrome virus (WSSV is a major pathogen in shrimp aquaculture. RNA interference (RNAi is a promising tool against viral infections. Previous works with RNAi showed different antiviral efficacies depending on the silenced gene. This work evaluated the antiviral efficacy of double-stranded (ds RNA against two non-structural (orf89, wsv191 WSSV genes compared to structural (vp26, vp28 genes to inhibit an experimental WSSV infection. Gene orf89 encodes a putative regulatory protein and gene white spot virus (wsv191 encodes a nonspecific nuclease; whereas genes vp26 and vp28 encode envelope proteins, respectively. Molecules of dsRNA against each of the WSSV genes were intramuscularly injected (4 μg per shrimp into a group of shrimp 48 h before a WSSV challenge. The highest antiviral activity occurred with dsRNA against orf89, vp28 and vp26 (cumulative mortalities 10%, 10% and 21%, respectively. In contrast, the least effective treatment was wsv191 dsRNA (cumulative mortality 83%. All dead animals were WSSV-positive by one-step PCR, whereas reverse-transcription PCR of all surviving shrimp confirmed inhibition of virus replication. This study showed that dsRNA against WSSV genes orf89, vp28 and vp26 were highly effective to inhibit virus replication and suggest an essential role in WSSV infection. Non-structural WSSV genes such as orf89 can be used as novel targets to design therapeutic RNAi molecules against WSSV infection.

Vp130, a chloroviral surface protein that interacts with the host Chlorella cell wall

International Nuclear Information System (INIS)

Onimatsu, Hideki; Sugimoto, Ichiro; Fujie, Makoto; Usami, Shoji; Yamada, Takashi

2004-01-01

A protein, Vp130, that interacts with the host cell wall was isolated from Chlorovirus CVK2. From its peptide sequence, the gene for Vp130 was identified on the PBCV-1 genomic sequence as an ORF combining A140R and A145R. In Vp130, the N-terminus was somehow modified and the C-terminus was occupied by 23-26 tandem repeats of a PAPK motif. In the internal region, Vp130 contained seven repeats of 70-73 amino acids, each copy of which was separated by PAPK sequences. This protein was well conserved among NC64A viruses. A recombinant rVp130N protein formed in Escherichia coli was shown not only to bind directly to the host cell wall in vitro but also to specifically bind to the host cells, as demonstrated by fluorescence microscopy. Because externally added rVp130N competed with CVK2 to bind to host cells, Vp130 is most likely to be a host-recognizing protein on the virion

Mechanism for Coordinated RNA Packaging and Genome Replication by Rotavirus Polymerase VP1

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Lu, Xiaohui; McDonald, Sarah M.; Tortorici, M. Alejandra; Tao, Yizhi Jane; Vasquez-Del Carpio, Rodrigo; Nibert, Max L.; Patton, John T.; Harrison, Stephen C. (Harvard-Med); (NIH); (CH-Boston)

2009-04-08

Rotavirus RNA-dependent RNA polymerase VP1 catalyzes RNA synthesis within a subviral particle. This activity depends on core shell protein VP2. A conserved sequence at the 3' end of plus-strand RNA templates is important for polymerase association and genome replication. We have determined the structure of VP1 at 2.9 {angstrom} resolution, as apoenzyme and in complex with RNA. The cage-like enzyme is similar to reovirus {lambda}3, with four tunnels leading to or from a central, catalytic cavity. A distinguishing characteristic of VP1 is specific recognition, by conserved features of the template-entry channel, of four bases, UGUG, in the conserved 3' sequence. Well-defined interactions with these bases position the RNA so that its 3' end overshoots the initiating register, producing a stable but catalytically inactive complex. We propose that specific 3' end recognition selects rotavirus RNA for packaging and that VP2 activates the autoinhibited VP1/RNA complex to coordinate packaging and genome replication.

Identification and characterization of vp7 gene in Bombyx mori cytoplasmic polyhedrosis virus.

Science.gov (United States)

He, Lei; Hu, Xiaolong; Zhu, Min; Liang, Zi; Chen, Fei; Zhu, Liyuan; Kuang, Sulan; Cao, Guangli; Xue, Renyu; Gong, Chengliang

2017-09-05

The genome of Bombyx mori cytoplasmic polyhedrosis virus (BmCPV) contains 10 double stranded RNA segments (S1-S10). The segment 7 (S7) encodes 50kDa protein which is considered as a structural protein. The expression pattern and function of p50 in the virus life cycle are still unclear. In this study, the viral structural protein 7 (VP7) polyclonal antibody was prepared with immunized mouse to explore the presence of small VP7 gene-encoded proteins in Bombyx mori cytoplasmic polyhedrosis virus. The expression pattern of vp7 gene was investigated by its overexpression in BmN cells. In addition to VP7, supplementary band was identified with western blotting technique. The virion, BmCPV infected cells and midguts were also examined using western blotting technique. 4, 2 and 5 bands were detected in the corresponding samples, respectively. The replication of BmCPV genome in the cultured cells and midgut of silkworm was decreased by reducing the expression level of vp7 gene using RNA interference. In immunoprecipitation experiments, using a polyclonal antiserum directed against the VP7, one additional shorter band in BmCPV infected midguts was detected, and then the band was analyzed with mass spectrum (MS), the MS results showed thatone candidate interacted protein (VP7 voltage-dependent anion-selective channel-like isoform, VDAC) was identified from silkworm. We concluded that the novel viral product was generated with a leaky scanning mechanism and the VDAC may be an interacted protein with VP7. Copyright © 2017 Elsevier B.V. All rights reserved.

Quantitative light microscopic autoradiographic localization of binding sites labelled with [3H]vasopressin antagonist d(CH2)5Tyr(Me)VP in the rat brain, pituitary and kidney

International Nuclear Information System (INIS)

Leeuwen, F.W. van; Beek, E.M. van der; Heerikhuize, J.J. van; Wolters, P.; Meulen, G. van der; Wan, Yieh-Ping

1987-01-01

Binding sites for the vasopressin (VP) antagonist d(CH 2 ) 5 Tyr(Me)VP, were located in various brain areas (e.g. the lateral septum, amygdala, choroid plexus and nucleus of the solitary tract) using light microscopic autoradiography. A number of areas (e.g. suprachiasmatic and arcuate nucleus, pineal gland) which previously showed no VP binding were labelled in the present study. The olfactory nucleus and ventromedial hypothalamic nucleus were not labelled. It therefore appears that d(CH 2 ) 5 Tyr(Me)VP is capable of discriminating between VP and oxytocin binding sites and more sensitive means of detecting VP binding sites than VP alone. (Author)

SV40 late protein VP4 forms toroidal pores to disrupt membranes for viral release.

Science.gov (United States)

Raghava, Smita; Giorda, Kristina M; Romano, Fabian B; Heuck, Alejandro P; Hebert, Daniel N

2013-06-04

Nonenveloped viruses are generally released from the cell by the timely lysis of host cell membranes. SV40 has been used as a model virus for the study of the lytic nonenveloped virus life cycle. The expression of SV40 VP4 at later times during infection is concomitant with cell lysis. To investigate the role of VP4 in viral release and its mechanism of action, VP4 was expressed and purified from bacteria as a fusion protein for use in membrane disruption assays. Purified VP4 perforated membranes as demonstrated by the release of fluorescent markers encapsulated within large unilamellar vesicles or liposomes. Dynamic light scattering results revealed that VP4 treatment did not cause membrane lysis or change the size of the liposomes. Liposomes encapsulated with 4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-3-indacene-labeled streptavidin were used to show that VP4 formed stable pores in membranes. These VP4 pores had an inner diameter of 1-5 nm. Asymmetrical liposomes containing pyrene-labeled lipids in the outer monolayer were employed to monitor transbilayer lipid diffusion. Consistent with VP4 forming toroidal pore structures in membranes, VP4 induced transbilayer lipid diffusion or lipid flip-flop. Altogether, these studies support a central role for VP4 acting as a viroporin in the disruption of cellular membranes to trigger SV40 viral release by forming toroidal pores that unite the outer and inner leaflets of membrane bilayers.

[Biological characteristics of a chimeric rabies virus expressing canine parvovirus VP2 protein].

Science.gov (United States)

Niu, Xue-Feng; Liu, Xiao-Hui; Sun, Zhao-Jin; Shi, He-He; Chen, Jing; Jiang, Bido; Sun, Jing-Chen; Guo, Xiao-Feng

2009-09-01

To obtain a bivalence vaccine against canine rabies virus and canine parvovirus, a chimeric rabies virus expressing canine parvovirus VP2 protein was generated by the technique of reverse genetics. It was shown that the chimeric virus designated as HEP-Flury (VP2) grew well on BHK-21 cells and the VP2 gene could still be stably expressed after ten passages on BHK-21 cells. Experiments on the mice immunized with the chimeric virus HEP-Flury (VP2) demonstrated that specific antibodies against rabies virus and canine parvovirus were induced in immunized mice after vaccination with the live chimeric virus.

Fifth VP Fuel Conservation Quarterly Report (June 1982 - August 1982) Supplement

National Research Council Canada - National Science Library

1982-01-01

...; and number of engines on during taxi. This report contains detailed analysis on fuel consumption during the months of June-August, 1982 for VP squadrons participating in the VP Fuel Conservation Study, (Patrol Squadrons Forty-Nine, Five, Twenty-Four, Fifty-Six and Sixteen).

The Imperative Sentence Pattern “Vp+tsau” in the Mengjin Dialect

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Peng Lanyu

2017-10-01

Full Text Available In the Mengjin dialect, the imperative sentence “Vp+ʦau (‘go’” is a common sentence pattern, which has six major structural forms. The basic structure is “tɕhy412(‘go’+(Np+Vp+ʦau (‘go’”, and “ʦau” in this form is a marginalized directional verb. Containing the stronger power factor, the imperative sentence “Vp+ʦau” is often used in the relationship between the superior and the subordinate, which has a high degree of colloquialism. Nevertheless, the power factor can be ignored on condition that the relationship between the two parties of the communication is very close. Due to the division of the ancient Chinese political jurisdictions and the influence of population migration, the imperative “Vp+ʦau” is widely distributed. Two different imperative sentences being merged and weakened, this sentence pattern comes into being.

Strategic Self-Presentation of Women in STEM

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Alexandra Garr-Schultz

2018-01-01

Full Text Available Despite a plethora of initiatives and a surge of research activity, women remain under-represented in science, technology, engineering, and mathematics (STEM disciplines (National Science Foundation 2017. While much research has focused on ways to recruit women into these disciplines, less work has explored the strategies women use to navigate these contexts once they have entered. In a set of two experimental studies, we investigate women’s potential response strategies to the well-documented tension between female and STEM attributes in terms of individual self-presentation. In Study 1 (N = 240, we examine whether female STEM professionals have different impression goals when introducing themselves to professional peers versus a group of other women. In Study 2 (N = 169, we extend our inquiry to include self-presentation behavior as well as intentions. Across studies, we find that female STEM professionals hold different impression goals based on the audience with whom and context in which they expect to interact. These intentions align with actual self-introduction behavior, as observed in written self-introductions. Tuning one’s self-presentation, however, leads participants to feel less authentic. This work highlights one way women in male-dominated STEM contexts may navigate and strategically communicate their female and STEM identities to others, as well as the personal implications of doing so.

Genetic diversity of G1P[8] rotavirus VP7 and VP8* antigens in Finland over a 20-year period: No evidence for selection pressure by universal mass vaccination with RotaTeq® vaccine.

Science.gov (United States)

Hemming, Maria; Vesikari, Timo

2013-10-01

Two live-attenuated oral vaccines (Rotarix™ and Rotateq®) against rotavirus gastroenteritis were licensed in 2006 and have been introduced into National Immunization Programs (NIPs) of several countries. Large scale use of rotavirus vaccines might cause antigenic pressure on circulating rotavirus types or lead to selection of new rotaviruses thus decreasing vaccine efficacy. We examined the nucleotide and amino acid sequences of the surface proteins VP7 and VP4 (cleaved to VP8(*) and VP5(*)) of a total of 108 G1P[8] rotavirus strains collected over a 20-year period from 1992, including the years 2006-2009 when rotavirus vaccine (mainly Rotarix™) was available, and the years 2009-2012 after implementation of RotaTeq® vaccine into the NIP of Finland. In G1 VP7 no changes at amino acid level were observed. In VP8(*) periodical fluctuation of the sublineage over the study period was found with multiple changes both at nucleotide and amino acid levels. Most amino acid changes were in the dominant antigenic epitopes of VP8(*). A change in VP8(*) sublineage occurred between 2008 and 2009, with a temporal correlation to the use of Rotarix™ up to 30% coverage in the period. In contrast, no antigenic changes in the VP8(*) protein appeared to be correlated to the exclusive use of RotaTeq® vaccine after 2009. Nevertheless, long-term surveillance of antigenic changes in VP4 and also VP7 proteins in wild-type rotavirus strains is warranted in countries with large scale use of the currently licensed live oral rotavirus vaccines. Copyright © 2013 Elsevier B.V. All rights reserved.

Capsid protein VP4 of human rhinovirus induces membrane permeability by the formation of a size-selective multimeric pore.

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Anusha Panjwani

2014-08-01

Full Text Available Non-enveloped viruses must deliver their viral genome across a cell membrane without the advantage of membrane fusion. The mechanisms used to achieve this remain poorly understood. Human rhinovirus, a frequent cause of the common cold, is a non-enveloped virus of the picornavirus family, which includes other significant pathogens such as poliovirus and foot-and-mouth disease virus. During picornavirus cell entry, the small myristoylated capsid protein VP4 is released from the virus, interacts with the cell membrane and is implicated in the delivery of the viral RNA genome into the cytoplasm to initiate replication. In this study, we have produced recombinant C-terminal histidine-tagged human rhinovirus VP4 and shown it can induce membrane permeability in liposome model membranes. Dextran size-exclusion studies, chemical crosslinking and electron microscopy demonstrated that VP4 forms a multimeric membrane pore, with a channel size consistent with transfer of the single-stranded RNA genome. The membrane permeability induced by recombinant VP4 was influenced by pH and was comparable to permeability induced by infectious virions. These findings present a molecular mechanism for the involvement of VP4 in cell entry and provide a model system which will facilitate exploration of VP4 as a novel antiviral target for the picornavirus family.

Expression and characterization of a novel truncated rotavirus VP4 for the development of a recombinant rotavirus vaccine.

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Li, Yijian; Xue, Miaoge; Yu, Linqi; Luo, Guoxing; Yang, Han; Jia, Lianzhi; Zeng, Yuanjun; Li, Tingdong; Ge, Shengxiang; Xia, Ningshao

2018-04-12

The outer capsid protein VP4 is an important target for the development of a recombinant rotavirus vaccine because it mediates the attachment and penetration of rotavirus. Due to the poor solubility of full-length VP4, VP8 was explored as candidate rotavirus vaccines in the past years. In previous studies, it has been found that the N-terminal truncated VP8 protein, VP8-1 (aa26-231), could be expressed in soluble form with improved immunogenicity compared to the core of VP8 (aa65-223). However, this protein stimulated only a weak immune response when aluminum hydroxide was used as an adjuvant. In addition, it should be noted that the protective efficacy of VP4 was higher than that of VP8 and VP5. In this study, it was found that when the N-terminal 25 amino acids were deleted, the truncated VP4 ∗ (aa26-476) containing VP8 and the stalk domain of VP5 could be expressed in soluble form in E. coli and purified to homogeneous trimers. Furthermore, the truncated VP4 could induce high titers of neutralizing antibodies when aluminum adjuvant was used and conferred high protective efficacy in reducing the severity of diarrhea and rotavirus shedding in stools in animal models. The immunogenicity of the truncated VP4 was significantly higher than that of VP8 ∗ and VP5 ∗ alone. Taken together, the truncated VP4 ∗ (aa26-476), with enhanced immunogenicity and immunoprotectivity, could be considered as a viable candidate for further development and has the potential to become a parenterally administered rotavirus vaccine. Copyright © 2018 Elsevier Ltd. All rights reserved.

The Ebola virus VP35 protein binds viral immunostimulatory and host RNAs identified through deep sequencing.

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Kari A Dilley

Full Text Available Ebola virus and Marburg virus are members of the Filovirdae family and causative agents of hemorrhagic fever with high fatality rates in humans. Filovirus virulence is partially attributed to the VP35 protein, a well-characterized inhibitor of the RIG-I-like receptor pathway that triggers the antiviral interferon (IFN response. Prior work demonstrates the ability of VP35 to block potent RIG-I activators, such as Sendai virus (SeV, and this IFN-antagonist activity is directly correlated with its ability to bind RNA. Several structural studies demonstrate that VP35 binds short synthetic dsRNAs; yet, there are no data that identify viral immunostimulatory RNAs (isRNA or host RNAs bound to VP35 in cells. Utilizing a SeV infection model, we demonstrate that both viral isRNA and host RNAs are bound to Ebola and Marburg VP35s in cells. By deep sequencing the purified VP35-bound RNA, we identified the SeV copy-back defective interfering (DI RNA, previously identified as a robust RIG-I activator, as the isRNA bound by multiple filovirus VP35 proteins, including the VP35 protein from the West African outbreak strain (Makona EBOV. Moreover, RNAs isolated from a VP35 RNA-binding mutant were not immunostimulatory and did not include the SeV DI RNA. Strikingly, an analysis of host RNAs bound by wild-type, but not mutant, VP35 revealed that select host RNAs are preferentially bound by VP35 in cell culture. Taken together, these data support a model in which VP35 sequesters isRNA in virus-infected cells to avert RIG-I like receptor (RLR activation.

The Ebola virus VP35 protein binds viral immunostimulatory and host RNAs identified through deep sequencing.

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Dilley, Kari A; Voorhies, Alexander A; Luthra, Priya; Puri, Vinita; Stockwell, Timothy B; Lorenzi, Hernan; Basler, Christopher F; Shabman, Reed S

2017-01-01

Ebola virus and Marburg virus are members of the Filovirdae family and causative agents of hemorrhagic fever with high fatality rates in humans. Filovirus virulence is partially attributed to the VP35 protein, a well-characterized inhibitor of the RIG-I-like receptor pathway that triggers the antiviral interferon (IFN) response. Prior work demonstrates the ability of VP35 to block potent RIG-I activators, such as Sendai virus (SeV), and this IFN-antagonist activity is directly correlated with its ability to bind RNA. Several structural studies demonstrate that VP35 binds short synthetic dsRNAs; yet, there are no data that identify viral immunostimulatory RNAs (isRNA) or host RNAs bound to VP35 in cells. Utilizing a SeV infection model, we demonstrate that both viral isRNA and host RNAs are bound to Ebola and Marburg VP35s in cells. By deep sequencing the purified VP35-bound RNA, we identified the SeV copy-back defective interfering (DI) RNA, previously identified as a robust RIG-I activator, as the isRNA bound by multiple filovirus VP35 proteins, including the VP35 protein from the West African outbreak strain (Makona EBOV). Moreover, RNAs isolated from a VP35 RNA-binding mutant were not immunostimulatory and did not include the SeV DI RNA. Strikingly, an analysis of host RNAs bound by wild-type, but not mutant, VP35 revealed that select host RNAs are preferentially bound by VP35 in cell culture. Taken together, these data support a model in which VP35 sequesters isRNA in virus-infected cells to avert RIG-I like receptor (RLR) activation.

Post V-P shunt surgical site EDH an uncommon complication: case report

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Garg Manish

2017-06-01

Full Text Available ventriculoparitoneal shunt is well established modality of treatment for hydrocephalous. Complication of v-p shunt are also mentioned in literature like shunt infection shunt migration etc [8]. Here we are describing a rare complication of vp shunt which barely mentioned in literature. A 22 yr male admitted with complain of headache & vomiting patient was diagnosed to have tubercular meningities with hydrocephalous. Patient planned for ventriculoparietoneal shunt surgery and vp shunt was done. On 3rd post-surgery day patient develop weakness in Left side of body. Urgent ncct head done which showed EDH at surgical site. Immediate craniotomy and evacuation of hematoma was done patient improved and discharged. Thus we are discussing the importance of meticulous surgery for v-p shunt, post op ct scan and treatment.

Synthesis and characterization of different immunogenic viral nanoconstructs from rotavirus VP6 inner capsid protein

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Bugli F

2014-05-01

Full Text Available Francesca Bugli,1 Valeria Caprettini,2 Margherita Cacaci,1 Cecilia Martini,1 Francesco Paroni Sterbini,1 Riccardo Torelli,1 Stefano Della Longa,3 Massimiliano Papi,4 Valentina Palmieri,4 Bruno Giardina,5 Brunella Posteraro,1 Maurizio Sanguinetti,1 Alessandro Arcovito5 1Istituto di Microbiologia, Università Cattolica del Sacro Cuore, 2Dipartimento di Fisica, Sapienza Università di Roma, Rome, 3Dipartimento di Medicina Clinica, Sanità Pubblica, Scienze della Vita e dell’Ambiente, Università dell’Aquila, L’Aquila, 4Istituto di Fisica, 5Istituto di Biochimica e Biochimica Clinica, Università Cattolica del Sacro Cuore, Rome, Italy Abstract: In order to deliver low-cost viral capsomeres from a large amount of soluble viral VP6 protein from human rotavirus, we developed and optimized a biotechnological platform in Escherichia coli. Specifically, three different expression protocols were compared, differing in their genetic constructs, ie, a simple native histidine-tagged VP6 sequence, VP6 fused to thioredoxin, and VP6 obtained with the newly described small ubiquitin-like modifier (SUMO fusion system. Our results demonstrate that the histidine-tagged protein does not escape the accumulation in the inclusion bodies, and that SUMO is largely superior to the thioredoxin-fusion tag in enhancing the expression and solubility of VP6 protein. Moreover, the VP6 protein produced according to the SUMO fusion tag displays well-known assembly properties, as observed in both transmission electron microscopy and atomic force microscopy images, giving rise to either VP6 trimers, 60 nm spherical virus-like particles, or nanotubes a few micron long. This different quaternary organization of VP6 shows a higher level of immunogenicity for the elongated structures with respect to the spheres or the protein trimers. Therefore, the expression and purification strategy presented here – providing a large amount of the viral capsid protein in the native

Head and neck angiography at 70 kVp with a third-generation dual-source CT system in patients: comparison with 100 kVp

Energy Technology Data Exchange (ETDEWEB)

Chen, Yu; Zhang, Xiaobo; Xue, Huadan; Zhu, Yuanli; Wang, Yun; Li, Yumei; Zhang, Zhuhua; Jin, Zhengyu [Chinese Academy of Medical Sciences, Department of Radiology, Peking Union Medical College Hospital, Beijing (China)

2017-11-15

This study was conducted in order to evaluate the image quality of 70 kVp and 25 mL contrast medium (CM) volume for head and neck computed tomographic angiography (CTA) and assess the diagnostic accuracy for arterial stenosis. Fifty patients were prospectively divided into two groups randomly: group A (n = 25), 70 kVp with 25 mL CM, and group B (n = 25), 100 kVp with 40 mL CM. CT attenuation values, noise, signal-to-noise ratio (SNR), and contrast-to-noise ratio (CNR) of the shoulder, neck, and cerebral arteries were measured for objective image quality. Subjective image quality of the shoulder and cerebral arteries was also evaluated. For patients undergoing digital subtracted angiography (DSA), diagnostic accuracy of CTA was assessed with DSA as reference standard. The SNRs of the shoulder, neck, and cerebral arteries in group A were higher than those in group B (P < 0.05). The CNRs of the shoulder and neck arteries in group A were higher than those in group B (P < 0.05). There was no significant difference in subjective image quality of arteries between group A and group B (P > 0.05). The accuracy was noted as 94.0% (156/166) in group A and 97.1% (134/138) in group B for ≥ 50% stenosis. The accuracy of intracranial arterial stenosis was lower than that of extracranial arterial stenosis in group A. The radiation dose of group A was significantly decreased by 56% than that of group B. Head and neck CTA at 70 kVp using 25 mL CM can obtain diagnostic image quality with lower radiation dose while maintaining high accuracy in detecting the arterial stenosis compared with the 100-kVp and 40-mL CM. (orig.)

Chaperone-Mediated Autophagy Protein BAG3 Negatively Regulates Ebola and Marburg VP40-Mediated Egress.

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Liang, Jingjing; Sagum, Cari A; Bedford, Mark T; Sidhu, Sachdev S; Sudol, Marius; Han, Ziying; Harty, Ronald N

2017-01-01

Ebola (EBOV) and Marburg (MARV) viruses are members of the Filoviridae family which cause outbreaks of hemorrhagic fever. The filovirus VP40 matrix protein is essential for virus assembly and budding, and its PPxY L-domain motif interacts with WW-domains of specific host proteins, such as Nedd4 and ITCH, to facilitate the late stage of virus-cell separation. To identify additional WW-domain-bearing host proteins that interact with VP40, we used an EBOV PPxY-containing peptide to screen an array of 115 mammalian WW-domain-bearing proteins. Using this unbiased approach, we identified BCL2 Associated Athanogene 3 (BAG3), a member of the BAG family of molecular chaperone proteins, as a specific VP40 PPxY interactor. Here, we demonstrate that the WW-domain of BAG3 interacts with the PPxY motif of both EBOV and MARV VP40 and, unexpectedly, inhibits budding of both eVP40 and mVP40 virus-like particles (VLPs), as well as infectious VSV-EBOV recombinants. BAG3 is a stress induced protein that regulates cellular protein homeostasis and cell survival through chaperone-mediated autophagy (CMA). Interestingly, our results show that BAG3 alters the intracellular localization of VP40 by sequestering VP40 away from the plasma membrane. As BAG3 is the first WW-domain interactor identified that negatively regulates budding of VP40 VLPs and infectious virus, we propose that the chaperone-mediated autophagy function of BAG3 represents a specific host defense strategy to counteract the function of VP40 in promoting efficient egress and spread of virus particles.

Chaperone-Mediated Autophagy Protein BAG3 Negatively Regulates Ebola and Marburg VP40-Mediated Egress.

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Jingjing Liang

2017-01-01

Full Text Available Ebola (EBOV and Marburg (MARV viruses are members of the Filoviridae family which cause outbreaks of hemorrhagic fever. The filovirus VP40 matrix protein is essential for virus assembly and budding, and its PPxY L-domain motif interacts with WW-domains of specific host proteins, such as Nedd4 and ITCH, to facilitate the late stage of virus-cell separation. To identify additional WW-domain-bearing host proteins that interact with VP40, we used an EBOV PPxY-containing peptide to screen an array of 115 mammalian WW-domain-bearing proteins. Using this unbiased approach, we identified BCL2 Associated Athanogene 3 (BAG3, a member of the BAG family of molecular chaperone proteins, as a specific VP40 PPxY interactor. Here, we demonstrate that the WW-domain of BAG3 interacts with the PPxY motif of both EBOV and MARV VP40 and, unexpectedly, inhibits budding of both eVP40 and mVP40 virus-like particles (VLPs, as well as infectious VSV-EBOV recombinants. BAG3 is a stress induced protein that regulates cellular protein homeostasis and cell survival through chaperone-mediated autophagy (CMA. Interestingly, our results show that BAG3 alters the intracellular localization of VP40 by sequestering VP40 away from the plasma membrane. As BAG3 is the first WW-domain interactor identified that negatively regulates budding of VP40 VLPs and infectious virus, we propose that the chaperone-mediated autophagy function of BAG3 represents a specific host defense strategy to counteract the function of VP40 in promoting efficient egress and spread of virus particles.

Molecular cloning and recombinant expression of the VP28 carboxyl-terminal hydrophilic region from a brazilian white spot syndrome virus isolate

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Patricia Braunig

2011-04-01

Full Text Available In the present study, a fragment of the VP28 coding sequence from a Brazilian WSSV isolate (BrVP28 was cloned, sequenced and expressed in E. coli BL21(DE3 pLysS strain in order to produce the VP28 carboxyl-terminal hydrophilic region. The expression resulted in a protein of about 21 kDa, which was purified under denaturing conditions, resulting in a final highly purified BrVP28 preparation. The recombinant protein obtained can be used in several biotechnology applications, such as the production of monoclonal antibodies which could be used in the development of diagnostic tools as well as in the studies on the characterization of white spot syndrome virus (WSSV isolated in Brazil.

kVp estimate intercomparison between Unfors XI, Radcal 4075 and a new CDTN multipurpose instrument

International Nuclear Information System (INIS)

Baptista Neto, A.T.; Oliveira, B.B.; Faria, L.O.

2015-01-01

In this work we compare the kVp estimate between CDTN multipurpose instrument, UnforsXI and Radcal 4075 meters under different combinations of voltage and filtration. The non-invasively measurements made using x-ray diagnostic and interventional radiology devices show similar tendencies to increase the kVp estimate when aluminum filters are placed in the path of the x-ray beam. The results reveal that the kVp estimate made by the CDTN multipurpose instrument is always satisfactory for highly filtered beam intensities. - Highlights: • We compare the kVp estimate between CDTN instrument and 2 different kVp meters. • The new CDTN multipurpose instrument performance was found to be satisfactory. • All instruments increase kVp estimative for increasing additional filtration. • They are suitable for quality control routines in x-ray diagnostic radiology

Differential Regulation of Interferon Responses by Ebola and Marburg Virus VP35 Proteins

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Edwards, Megan R.; Liu, Gai; Mire, Chad E.; Sureshchandra, Suhas; Luthra, Priya; Yen, Benjamin; Shabman, Reed S.; Leung, Daisy W.; Messaoudi, Ilhem; Geisbert, Thomas W.; Amarasinghe, Gaya K.; Basler, Christopher F.

2016-02-11

Suppression of innate immune responses during filoviral infection contributes to disease severity. Ebola (EBOV) and Marburg (MARV) viruses each encode a VP35 protein that suppresses RIG-I-like receptor signaling and interferon-α/β (IFN-α/β) production by several mechanisms, including direct binding to double stranded RNA (dsRNA). Here, we demonstrate that in cell culture, MARV infection results in a greater upregulation of IFN responses as compared to EBOV infection. This correlates with differences in the efficiencies by which EBOV and MARV VP35s antagonize RIG-I signaling. Furthermore, structural and biochemical studies suggest that differential recognition of RNA elements by the respective VP35 C-terminal IFN inhibitory domain (IID) rather than affinity for RNA by the respective VP35s is critical for this observation. Our studies reveal functional differences in EBOV versus MARV VP35 RNA binding that result in unexpected differences in the host response to deadly viral pathogens.

[Genetic variation analysis of canine parvovirus VP2 gene in China].

Science.gov (United States)

Yi, Li; Cheng, Shi-Peng; Yan, Xi-Jun; Wang, Jian-Ke; Luo, Bin

2009-11-01

To recognize the molecular biology character, phylogenetic relationship and the state quo prevalent of Canine parvovirus (CPV), Faecal samnples from pet dogs with acute enteritis in the cities of Beijing, Wuhan, and Nanjing were collected and tested for CPV by PCR and other assay between 2006 and 2008. There was no CPV to FPV (MEV) variation by PCR-RFLP analysis in all samples. The complete ORFs of VP2 genes were obtained by PCR from 15 clinical CPVs and 2 CPV vaccine strains. All amplicons were cloned and sequenced. Analysis of the VP2 sequences showed that clinical CPVs both belong to CPV-2a subtype, and could be classified into a new cluster by amino acids contrasting which contains Tyr-->Ile (324) mutation. Besides the 2 CPV vaccine strains belong to CPV-2 subtype, and both of them have scattered variation in amino acids residues of VP2 protein. Construction of the phylogenetic tree based on CPV VP2 sequence showed these 15 CPV clinical strains were in close relationship with Korea strain K001 than CPV-2a isolates in other countries at early time, It is indicated that the canine parvovirus genetic variation was associated with location and time in some degree. The survey of CPV capsid protein VP2 gene provided the useful information for the identification of CPV types and understanding of their genetic relationship.

Mutations Abrogating VP35 Interaction with Double-Stranded RNA Render Ebola Virus Avirulent in Guinea Pigs

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Prins, Kathleen C.; Delpeut, Sebastien; Leung, Daisy W.; Reynard, Olivier; Volchkova, Valentina A.; Reid, St. Patrick; Ramanan, Parameshwaran; Cárdenas, Washington B.; Amarasinghe, Gaya K.; Volchkov, Viktor E.; Basler, Christopher F. (CNRS-INSERM); (Mount Sinai Hospital); (LB-Ecuador); (Iowa State)

2010-10-11

Ebola virus (EBOV) protein VP35 is a double-stranded RNA (dsRNA) binding inhibitor of host interferon (IFN)-{alpha}/{beta} responses that also functions as a viral polymerase cofactor. Recent structural studies identified key features, including a central basic patch, required for VP35 dsRNA binding activity. To address the functional significance of these VP35 structural features for EBOV replication and pathogenesis, two point mutations, K319A/R322A, that abrogate VP35 dsRNA binding activity and severely impair its suppression of IFN-{alpha}/{beta} production were identified. Solution nuclear magnetic resonance (NMR) spectroscopy and X-ray crystallography reveal minimal structural perturbations in the K319A/R322A VP35 double mutant and suggest that loss of basic charge leads to altered function. Recombinant EBOVs encoding the mutant VP35 exhibit, relative to wild-type VP35 viruses, minimal growth attenuation in IFN-defective Vero cells but severe impairment in IFN-competent cells. In guinea pigs, the VP35 mutant virus revealed a complete loss of virulence. Strikingly, the VP35 mutant virus effectively immunized animals against subsequent wild-type EBOV challenge. These in vivo studies, using recombinant EBOV viruses, combined with the accompanying biochemical and structural analyses directly correlate VP35 dsRNA binding and IFN inhibition functions with viral pathogenesis. Moreover, these studies provide a framework for the development of antivirals targeting this critical EBOV virulence factor.

Construction and characterization of human rotavirus recombinant VP8* subunit parenteral vaccine candidates.

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Wen, Xiaobo; Cao, Dianjun; Jones, Ronald W; Li, Jianping; Szu, Shousun; Hoshino, Yasutaka

2012-09-21

Two currently licensed live oral rotavirus vaccines (Rotarix® and RotaTeq®) are highly efficacious against severe rotavirus diarrhea. However, the efficacy of such vaccines in selected low-income African and Asian countries is much lower than that in middle or high-income countries. Additionally, these two vaccines have recently been associated with rare case of intussusception in vaccinated infants. We developed a novel recombinant subunit parenteral rotavirus vaccine which may be more effective in low-income countries and also avert the potential problem of intussusception. Truncated recombinant VP8* (ΔVP8*) protein of human rotavirus strain Wa P[8], DS-1 P[4] or 1076 P[6] expressed in Escherichia coli was highly soluble and was generated in high yield. Guinea pigs hyperimmunized intramuscularly with each of the ΔVP8* proteins (i.e., P[8], P[4] or P[6]) developed high levels of homotypic as well as variable levels of heterotypic neutralizing antibodies. Moreover, the selected ΔVP8* proteins when administered to mice at a clinically relevant dosage, route and schedule, elicited high levels of serum anti-VP8* IgG and/or neutralizing antibodies. Our data indicated that the ΔVP8* proteins may be a plausible additional candidate as new parenteral rotavirus vaccines. Published by Elsevier Ltd.

Molecular cloning and sequence analysis of VP6 gene of giant ...

African Journals Online (AJOL)

Jane

2011-10-24

Oct 24, 2011 ... G), and the major structural protein of inner capsid particles (ICP), and also specific antigen of mucosa immunization that mediate specific immunological reaction. In this report, sequence analysis of VP6 gene of giant panda rotavirus was carried out. Full-length VP6 gene encoding for ICP of giant panda.

Ebola virus VP24 interacts with NP to facilitate nucleocapsid assembly and genome packaging.

Science.gov (United States)

Banadyga, Logan; Hoenen, Thomas; Ambroggio, Xavier; Dunham, Eric; Groseth, Allison; Ebihara, Hideki

2017-08-09

Ebola virus causes devastating hemorrhagic fever outbreaks for which no approved therapeutic exists. The viral nucleocapsid, which is minimally composed of the proteins NP, VP35, and VP24, represents an attractive target for drug development; however, the molecular determinants that govern the interactions and functions of these three proteins are still unknown. Through a series of mutational analyses, in combination with biochemical and bioinformatics approaches, we identified a region on VP24 that was critical for its interaction with NP. Importantly, we demonstrated that the interaction between VP24 and NP was required for both nucleocapsid assembly and genome packaging. Not only does this study underscore the critical role that these proteins play in the viral replication cycle, but it also identifies a key interaction interface on VP24 that may serve as a novel target for antiviral therapeutic intervention.

Bioinformatic prediction of polymerase elements in the rotavirus VP1 protein

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RODRIGO VÁSQUEZ-DEL CARPIÓ

2006-01-01

Full Text Available Rotaviruses are the major cause of acute gastroenteritis in infants world-wide. The genome consists of eleven double stranded RNA segments. The major segment encodes the structural protein VP1, the viral RNA-dependent RNA polymerase (RdRp, which is a minor component of the viral inner core. This study is a detailed bioinformatic assessment of the VP1 sequence. Using various methods we have identified canonical motifs within the VP1 sequence which correspond to motifs previously identified within RdRps of other positive strand, double-strand RNA viruses. The study also predicts an overall structural conservation in the middle region that may correspond to the palm subdomain and part of the fingers and thumb subdomains, which comprise the polymerase core of the protein. Based on this analysis, we suggest that the rotavirus replicase has the minimal elements to function as an RNA-dependent RNA polymerase. VP1, besides having common RdRp features, also contains large unique regions that might be responsible for characteristic features observed in the Reoviridae family

dsRNA binding characterization of full length recombinant wild type and mutants Zaire ebolavirus VP35.

Science.gov (United States)

Zinzula, Luca; Esposito, Francesca; Pala, Daniela; Tramontano, Enzo

2012-03-01

The Ebola viruses (EBOVs) VP35 protein is a multifunctional major virulence factor involved in EBOVs replication and evasion of the host immune system. EBOV VP35 is an essential component of the viral RNA polymerase, it is a key participant of the nucleocapsid assembly and it inhibits the innate immune response by antagonizing RIG-I like receptors through its dsRNA binding function and, hence, by suppressing the host type I interferon (IFN) production. Insights into the VP35 dsRNA recognition have been recently revealed by structural and functional analysis performed on its C-terminus protein. We report the biochemical characterization of the Zaire ebolavirus (ZEBOV) full-length recombinant VP35 (rVP35)-dsRNA binding function. We established a novel in vitro magnetic dsRNA binding pull down assay, determined the rVP35 optimal dsRNA binding parameters, measured the rVP35 equilibrium dissociation constant for heterologous in vitro transcribed dsRNA of different length and short synthetic dsRNA of 8bp, and validated the assay for compound screening by assessing the inhibitory ability of auryntricarboxylic acid (IC(50) value of 50μg/mL). Furthermore, we compared the dsRNA binding properties of full length wt rVP35 with those of R305A, K309A and R312A rVP35 mutants, which were previously reported to be defective in dsRNA binding-mediated IFN inhibition, showing that the latter have measurably increased K(d) values for dsRNA binding and modified migration patterns in mobility shift assays with respect to wt rVP35. Overall, these results provide the first characterization of the full-length wt and mutants VP35-dsRNA binding functions. Copyright © 2012 Elsevier B.V. All rights reserved.

VP Market teeb rekordkasvu / Ralf-Martin Soe

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Soe, Ralf-Martin

2006-01-01

Ilmunud ka: Delovõje Vedomosti 26. apr. lk. 4. Leedu omanikele kuuluv T-Marketite poekett teeb Eestis rekordilist käibe kasvu. Diagramm. Vt. samas: Raimo Ülavere. VP Market annab konkurentidele silmad ette

[Immune Response of Recombinant Pseudorabies Virus rPRV-VP2 Expressing VP2 Gene of Porcine Parvovirus in Mice].

Science.gov (United States)

Fu, Pengfei; Pan, Xinlong; Han, Qiao; Yang, Xingwu; Zhu, Qianlei; Guo, Xiaoqing; Zhang, Yu; Chen, Hongying

2016-03-01

In order to develop a combined live vaccine that will be used to prevent against porcine parvovirus (PPV) and Pseudorabies virus (PRV) infection, the VP2 gene of PPV was inserted into the transfer vector plasmid pG to produce the recombinant plasmid pGVP2. The plasmid pGVP2 and the genome of PRV HB98 attenuated vaccine were transfected by using lipofectamine into swine testis cells for the homologous recombination. The recombinant virus rPRV-VP2 was purified by selection of green fluorescence plaques for five cycles. 6-week-old female Kunming mice were immunized intramuscularly with attenuated PRV parent HB98 strain, commercial inactivated vaccine against PPV, recombinant virus, DMEM culture solution. The injections were repeated with an equivalent dose after 2 weeks in all of the groups, and then challenged with the virulent PRV NY strain at 7 weeks after the first immunization. The recombinant virus rPRV-VP2 was successfully generated, and the recombinant virus could effectively elicite anti-PPV and PRV antibody and significant cellular immune response as indicated by anti-PPV ELISA and HI, PRV-neutralizing assay and flow cytometry. The challenge assay indicated that recombinant virus could protect the mice against the virulent PRV challenge. These results demonstrated that the recombinant virus can be a candidate recombinant vaccine strain for the prevention of PRV and PPV.

VP6-SUMO Self-Assembly as Nanocarriers for Gastrointestinal Delivery

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V. Palmieri

2015-01-01

Full Text Available High proteolytic degradation and poor absorption through epithelial barriers are major challenges to successful oral delivery of therapeutics. Nanoparticle platforms can enhance drug stability and extend the residence time in gastrointestinal (GI tract. However, drug delivery systems are often inactivated in acidic environment of stomach or suffer poor absorption from intestinal cells due to the mucus layer. To overcome these issues we developed a drug delivery system constituted by a protein construct made by a Rotavirus capsid protein (VP6 and the small ubiquitin-like modifier SUMO. This chimeric construct allows specificity towards intestinal cells, the Rotavirus natural target, combined by an enhanced stability given by the eukaryotic protein transporter SUMO. Furthermore SUMO can act as a molecular switch that facilitates import/export of its ligand to the nucleus, the hypersensitive subcellular site target of many cell killing therapies. In this paper we show that SUMO-VP6 constructs self-assembly into stable nanocarriers. SUMO-VP6 nanocarriers display ideal features for drug delivery: a small size and high monodispersity, a high stability in different pH conditions and a high uptake in the nuclear and cytoplasmic compartment of intestinal cells. These features make SUMO-VP6 nanocarriers a promising novel system for oral delivery of poorly soluble drugs.

VP Market Eesti tööseadustega pahuksis / Merike Mikk

Index Scriptorium Estoniae

Mikk, Merike, 1973-

2004-01-01

Leedu jaekaubanduskontserni VP Market endiste töötajate sõnul on ettevõte rikkunud Eestis kehtivaid tööseadusi töölepingute sõlmimisel, mille tõttu on Tartu ja Tallinna töövaidluskomisjonidele esitatud lahendamiseks 14 avaldust. Vt. samas: Leedu poekett kasvab jõuliselt; T-Marketid saanud kaks ettekirjutust; VP Marketi kommentaar artiklile; Säästumarketis lisatasu kord kirjas; Intervjuu endise T-Marketi juhataja Tessa Kütimetsaga

The relative biological effectiveness of 60Co γ-rays, 55 kVp X-rays, 250 kVp X-rays, and 11 MeV electrons at low doses

International Nuclear Information System (INIS)

Spadinger, I.; Palcic, B.

1992-01-01

The RBE of selected low-LET radiation modalities (55 kVp X- rays, 250 kVp X-rays, 60 Co γ-rays, and 11 MeV electrons) was investigated for survival of two cell lines (V79 and CHO). Detailed measurements were made in the 0 to 3 Gy dose range using an image cytometry device to accurately determine the number of cells assayed at each dose point. Data were also collected in the high dose range (0 to 10 Gy) using conventional counting and plating techniques. RBE values (#+- #1 SE) varied from 1.0±0.07 (V79 cells) and 1.2± 0.05 (CHO cells) at high doses to 1.3±0.07 (V79) and 1.4±0.1 (CHO) at low doses for 55 kVp X-rays, from 1.1±0.05 (V79) and 1.1±0.04 (CHO) at high doses to 1.1±0.06 (V79) and 1.2±0.2 (CHO) at low doses for 250 kVp X-rays, and from 1.1±0.08 (V79) and 1.0±0.04 (CHO) at high doses to 1.0±0.06 (V79) and 0.9±0.1 (CHO) at low doses for 11 MeV electrons. Only the low and high dose RBEs for 55 kVp X-rays relative to 60 Co γ-rays were significantly different. (author)

The use of an in vitro microneutralization assay to evaluate the potential of recombinant VP5 protein as an antigen for vaccinating against Grass carp reovirus

Directory of Open Access Journals (Sweden)

Xu Dan

2011-03-01

Full Text Available Abstract Background Grass carp reovirus (GCRV is the causative pathogen of grass carp hemorrhagic disease, one of the major diseases damaging grass carp Ctenopharyngon idellus breeding industry in China. Prevention and control of the disease is impeded largely due to the lack of research in economic subunit vaccine development. This study aimed to evaluate the potential of viral outer shell protein VP5 as subunit vaccine. Methods The vp5 gene was isolated from the viral genome through RT-PCR and genetically engineered to express the recombinant VP5 protein in E coli. The viral origin of the recombinant protein was confirmed by Western blot analysis with a monoclonal antibody against viral VP5 protein. Polyclonal antibody against the recombinant VP5 protein was prepared from mice. A microneutralization assay was developed to test its neutralizing ability against GCRV infection in cell culture. Results The GST-VP5 fusion protein (rVP5 was produced from E. Coli with expected molecular weight of 90 kDa. The protein was purified and employed to prepare anti-VP5 polyclonal antibody from mice. The anti-VP5 antibody was found to neutralize GCRV through in vitro microneutralization assay and viral progeny quantification analysis. Conclusions The present study showed that the viral VP5 protein was involved in viral infection and bacterially-expressed VP5 could be suitable for developing subunit vaccine for the control of GCRV infection.

Mathematically Gifted High School Students' Approaches to Developing Visual Proofs (VP) and Preliminary Ideas about VP

Science.gov (United States)

Ugurel, Isikhan; Morali, H. Sevgi; Karahan, Ozge; Boz, Burcak

2016-01-01

The purpose of this study is to describe the procedure and examples of visual proofs (VP-or proof without words) developed by gifted mathematics secondary school students after their experiences. The participants of this study are three male 9th grade students enrolled in a private science high school. In the first stage of the research a briefing…

Immunogenicity and protective efficacy of rotavirus VP8* fused to cholera toxin B subunit in a mouse model.

Science.gov (United States)

Xue, Miaoge; Yu, Linqi; Jia, Lianzhi; Li, Yijian; Zeng, Yuanjun; Li, Tingdong; Ge, Shengxiang; Xia, Ningshao

2016-11-01

In attempts to develop recombinant subunit vaccines against rotavirus disease, it was previously shown that the N-terminal truncated VP8* protein, VP8-1 (aa26-231), is a good vaccine candidate when used for immunization in combination with Freund's adjuvant. However, this protein stimulated only weak immune response when aluminum hydroxide was used as an adjuvant. In this study, the nontoxic B subunit of cholera toxin (CTB) was employed as intra-molecular adjuvant to improve the immunogenicity of VP8-1. Both, the N-terminal and C-terminal fusion proteins, were purified to homogeneity, at which stage they formed pentamers, and showed significantly higher immunogenicity and protective efficacy than a VP8-1/aluminum hydroxide mixture in a mouse model. Compared to VP8-1-CTB, CTB-VP8-1 showed higher binding activity to both, GM1 and the conformation sensitive neutralizing monoclonal antibodies specific to VP8. More importantly, CTB-VP8-1 elicited higher titers of neutralizing antibodies and conferred higher protective efficacy than VP8-1-CTB. Therefore, the protein CTB-VP8-1, with enhanced immunogenicity and immunoprotectivity, could be considered as a viable candidate for further development of an alternative, replication-incompetent, parenterally administered vaccine against rotavirus disease.

Antigenicity analysis of human parvovirus B19-VP1u protein in the induction of anti-phospholipid syndrome.

Science.gov (United States)

Lin, Chun-Yu; Chiu, Chun-Ching; Cheng, Ju; Lin, Chia-Yun; Shi, Ya-Fang; Tsai, Chun-Chou; Tzang, Bor-Show; Hsu, Tsai-Ching

2018-01-01

Mounting evidence suggests a connection between human parvovirus B19 (B19) and autoimmune diseases, and especially an association between the B19-VP1 unique region (VP1u) and anti-phospholipid syndrome (APS). However, little is known about the antigenicity of B19-VP1u in the induction of APS-like syndrome. To elucidate the antigenicity of B19-VP1u in the induction of APS, N-terminal truncated B19-VP1u (tVP1u) proteins were prepared to immunize Balb/c mice to generate antibodies against B19-tVP1u proteins. The secreted phospholipase A2 (sPLA2) activities and binding specificity of mice anti-B19-tVP1u antibodies with cardiolipin (CL) and beta-2-glycoprotein I (β2GPI) were evaluated by performing immunoblot, ELISA and absorption experiments. A mice model of passively induced APS was adopted. Although sPLA2 activities were identified in all B19-tVP1u proteins, only amino acid residues 61-227 B19-tVP1u exhibited a higher sPLA2 activity. Autoantibodies against CL and β2GPI exhibited binding activities with all B19-tVP1u proteins. IgG that was purified from mice that had been immunized with amino acid residues 21-227 to 121-227 B19-tVP1u proteins exhibited significantly higher binding activity with CL. IgG that was purified from mice that had been immunized with amino acid residues 21-227, 31-227, 82-227 and 91-227 B19-tVP1u proteins exhibited significantly higher binding activity with β2GPI. Accordingly, significantly higher binding inhibition of CL was detected in the presence of amino acid residues 61-227 and 101-227 B19-tVP1u. Significantly higher binding inhibition of β2GPI was detected in the presence of amino acid residues 21-227, 31-227, 82-227 and 91-227 B19-tVP1u. The mice that received amino acid residues 31-227 or 61-227 anti-tB19-VP1u IgG revealed significant thrombocytopenia and those that received amino acid residues 21-227, 31-227, 61-227, 71-227, 82-227, 91-227, 101-227 or 114-227 anti-tB19-VP1u IgG exhibited significantly prolonged aPTT. These

Expression, purification, crystallization and preliminary X-ray studies of the Ebola VP35 interferon inhibitory domain

International Nuclear Information System (INIS)

Leung, Daisy W.; Ginder, Nathaniel D.; Nix, Jay C.; Basler, Christopher F.; Honzatko, Richard B.; Amarasinghe, Gaya K.

2009-01-01

Native and selenomethionine-labeled crystals of Ebola VP35 interferon inhibitory domain were obtained by the hanging-drop vapor-diffusion method. Ebola VP35 is a multifunctional protein that is important for host immune suppression and pathogenesis. VP35 contains an N-terminal oligomerization domain and a C-terminal interferon inhibitory domain (IID). Mutations within the VP35 IID result in loss of host immune suppression. Here, efforts to crystallize recombinantly overexpressed VP35 IID that was purified from Escherichia coli are described. Native and selenomethionine-labeled crystals belonging to the orthorhombic space group P2 1 2 1 2 1 were obtained by the hanging-drop vapor-diffusion method and diffraction data were collected at the ALS synchrotron

Supramolecular Assembly of Gold Nanoparticles in PS-b-P2VP Diblock Copolymers via Hydrogen Bonding

Science.gov (United States)

Jang, Se Gyu; Hawker, Craig J.; Kramer, Edward J.

2011-03-01

We report a simple route to control the spatial distribution of Au nanoparticles (Au-NPs) in PS- b -P2VP diblock copolymers using hydrogen bonding between P2VP and the hydroxyl-containing (PI-OH) units in PS- b -PIOH thiol-terminated ligands on Au-NP. End-functional thiol ligands of poly(styrene- b -1,2&3,4-isoprene-SH) are synthesized by anionic polymerization. After synthesis of Au-NPs, the inner PI block is hydroxylated by hydroboration and the resulting micelle-like Au-NPs consist of a hydrophobic PS outer brush and a hydrophilic inner PI-OH block. The influence of the hydroxyl groups is significant with strong segregation being observed to the PS/P2VP interface and then to the P2VP domain of lamellar-forming PS-b-P2VP diblock copolymers as the length of the PI-OH block is increased. The strong hydrogen bonding between nanoparticle block copolymer ligands and the P2VP block allows the Au-NPs to be incorporated within the P2VP domain to high Au--NP volume fractions ϕp without macrophase separation, driving transitions from lamellar to bicontinuous morphologies as ϕp increases.

Expression of of VP60 gene from RHDV YL strain under control of ...

African Journals Online (AJOL)

zdx

2012-06-19

Jun 19, 2012 ... So far, several plant species suitable for the expression of recombinant proteins with ... plant transfer vector including the VP60 gene driven by .... VP60 protein protects against rabbit hemorrhagic disease virus. J. Virol. 73(5): ...

Tandem truncated rotavirus VP8* subunit protein with T cell epitope as non-replicating parenteral vaccine is highly immunogenic.

Science.gov (United States)

Wen, Xiaobo; Cao, Dianjun; Jones, Ronald W; Hoshino, Yasutaka; Yuan, Lijuan

2015-01-01

The two currently available live oral rotavirus vaccines, Rotarix(®) and RotaTeq(®), are highly efficacious in the developed countries. However, the efficacy of such vaccines in resource deprived countries in Africa and Southeast Asia is low. We reported previously that a bacterially-expressed rotavirus P2-P[8] ΔVP8* subunit vaccine candidate administered intramuscularly elicited high-titers of neutralizing antibodies in guinea pigs and mice and significantly shortened the duration of diarrhea in neonatal gnotobiotic pigs upon oral challenge with virulent human rotavirus Wa strain. To further improve its vaccine potential and provide wider coverage against rotavirus strains of global and regional epidemiologic importance, we constructed 2 tandem recombinant VP8* proteins, P2-P[8] ΔVP8*-P[8] ΔVP8* and P2-P[8] ΔVP8*-P[6] ΔVP8* based on Escherichia coli expression system. The two resulting recombinant tandem proteins were highly soluble and P2-P[8] ΔVP8*-P[8] ΔVP8* was generated with high yield. Moreover, guinea pigs immunized intramuscularly by 3 doses of the P2-P[8] ΔVP8*-P[8] ΔVP8* or P2-P[8] ΔVP8*-P[6] ΔVP8* vaccine with aluminum phosphate adjuvant developed high titers of homotypic and heterotypic neutralizing antibodies against human rotaviruses bearing G1-G4, G8, G9 and G12 with P[8], P[4] or P[6] combination. The results suggest that these 2 subunit vaccines in monovalent or bivalent formulation can provide antigenic coverage to almost all the rotavirus G (VP7) types and major P (VP4) types of global as well as regional epidemiologic importance.

Crystallization and preliminary X-ray analysis of Ebola VP35 interferon inhibitory domain mutant proteins

International Nuclear Information System (INIS)

Leung, Daisy W.; Borek, Dominika; Farahbakhsh, Mina; Ramanan, Parameshwaran; Nix, Jay C.; Wang, Tianjiao; Prins, Kathleen C.; Otwinowski, Zbyszek; Honzatko, Richard B.; Helgeson, Luke A.; Basler, Christopher F.; Amarasinghe, Gaya K.

2010-01-01

Three mutant forms of Ebola VP35 interferon inhibitory domain were crystallized in three different space groups. VP35 is one of seven structural proteins encoded by the Ebola viral genome and mediates viral replication, nucleocapsid formation and host immune suppression. The C-terminal interferon inhibitory domain (IID) of VP35 is critical for dsRNA binding and interferon inhibition. The wild-type VP35 IID structure revealed several conserved residues that are important for dsRNA binding and interferon antagonism. Here, the expression, purification and crystallization of recombinant Zaire Ebola VP35 IID mutants R312A, K319A/R322A and K339A in space groups P6 1 22, P2 1 2 1 2 1 and P2 1 , respectively, are described. Diffraction data were collected using synchrotron sources at the Advanced Light Source and the Advanced Photon Source

A Single Amino Acid Change in the Marburg Virus Matrix Protein VP40 Provides a Replicative Advantage in a Species-Specific Manner

Science.gov (United States)

Koehler, Alexander; Kolesnikova, Larissa; Welzel, Ulla; Schudt, Gordian; Herwig, Astrid

2015-01-01

ABSTRACT Marburg virus (MARV) induces severe hemorrhagic fever in humans and nonhuman primates but only transient nonlethal disease in rodents. However, sequential passages of MARV in rodents boosts infection leading to lethal disease. Guinea pig-adapted MARV contains one mutation in the viral matrix protein VP40 at position 184 (VP40D184N). The contribution of the D184N mutation to the efficacy of replication in a new host is unknown. In the present study, we demonstrated that recombinant MARV containing the D184N mutation in VP40 [rMARVVP40(D184N)] grew to higher titers than wild-type recombinant MARV (rMARVWT) in guinea pig cells. Moreover, rMARVVP40(D184N) displayed higher infectivity in guinea pig cells. Comparative analysis of VP40 functions indicated that neither the interferon (IFN)-antagonistic function nor the membrane binding capabilities of VP40 were affected by the D184N mutation. However, the production of VP40-induced virus-like particles (VLPs) and the recruitment of other viral proteins to the budding site was improved by the D184N mutation in guinea pig cells, which resulted in the higher infectivity of VP40D184N-induced infectious VLPs (iVLPs) compared to that of VP40-induced iVLPs. In addition, the function of VP40 in suppressing viral RNA synthesis was influenced by the D184N mutation specifically in guinea pig cells, thus allowing greater rates of transcription and replication. Our results showed that the improved viral fitness of rMARVVP40(D184N) in guinea pig cells was due to the better viral assembly function of VP40D184N and its lower inhibitory effect on viral transcription and replication rather than modulation of the VP40-mediated suppression of IFN signaling. IMPORTANCE The increased virulence achieved by virus passaging in a new host was accompanied by mutations in the viral genome. Analyzing how these mutations affect the functions of viral proteins and the ability of the virus to grow within new host cells helps in the understanding

A novel recombinant pseudorabies virus expressing parvovirus VP2 gene: Immunogenicity and protective efficacy in swine.

Science.gov (United States)

Chen, Yang; Guo, Wanzhu; Xu, Zhiwen; Yan, Qigui; Luo, Yan; Shi, Qian; Chen, Dishi; Zhu, Ling; Wang, Xiaoyu

2011-06-16

Porcine parvovirus (PPV) VP2 gene has been successfully expressed in many expression systems resulting in self-assembly of virus-like particles (VLPs) with similar morphology to the native capsid. Here, a pseudorabies virus (PRV) system was adopted to express the PPV VP2 gene. A recombinant PRV SA215/VP2 was obtained by homologous recombination between the vector PRV viral DNA and a transfer plasmid. Then recombinant virus was purified with plaque purification, and its identity confirmed by PCR amplification, Western blot and indirect immunofluorescence (IFA) analyses. Electronic microscopy of PRV SA215/VP2 confirmed self-assembly of both pseudorabies virus and VLPs from VP2 protein. Immunization of piglets with recombinant virus elicited PRV-specific and PPV-specific humoral immune responses and provided complete protection against a lethal dose of PRV challenges. Gilts immunized with recombinant viruses induced PPV-specific antibodies, and significantly reduced the mortality rate of (1 of 28) following virulent PPV challenge compared with the control (7 of 31). Furthermore, PPV virus DNA was not detected in the fetuses of recombinant virus immunized gilts. In this study, a recombinant PRV SA215/VP2 virus expressing PPV VP2 protein was constructed using PRV SA215 vector. The safety, immunogenicity, and protective efficacy of the recombinant virus were demonstrated in piglets and primiparous gilts. This recombinant PRV SA215/VP2 represents a suitable candidate for the development of a bivalent vaccine against both PRV and PPV infection.

The ebola virus interferon antagonist VP24 directly binds STAT1 and has a novel, pyramidal fold.

Science.gov (United States)

Zhang, Adrianna P P; Bornholdt, Zachary A; Liu, Tong; Abelson, Dafna M; Lee, David E; Li, Sheng; Woods, Virgil L; Saphire, Erica Ollmann

2012-02-01

Ebolaviruses cause hemorrhagic fever with up to 90% lethality and in fatal cases, are characterized by early suppression of the host innate immune system. One of the proteins likely responsible for this effect is VP24. VP24 is known to antagonize interferon signaling by binding host karyopherin α proteins, thereby preventing them from transporting the tyrosine-phosphorylated transcription factor STAT1 to the nucleus. Here, we report that VP24 binds STAT1 directly, suggesting that VP24 can suppress at least two distinct branches of the interferon pathway. Here, we also report the first crystal structures of VP24, derived from different species of ebolavirus that are pathogenic (Sudan) and nonpathogenic to humans (Reston). These structures reveal that VP24 has a novel, pyramidal fold. A site on a particular face of the pyramid exhibits reduced solvent exchange when in complex with STAT1. This site is above two highly conserved pockets in VP24 that contain key residues previously implicated in virulence. These crystal structures and accompanying biochemical analysis map differences between pathogenic and nonpathogenic viruses, offer templates for drug design, and provide the three-dimensional framework necessary for biological dissection of the many functions of VP24 in the virus life cycle.

The ebola virus interferon antagonist VP24 directly binds STAT1 and has a novel, pyramidal fold.

Directory of Open Access Journals (Sweden)

Adrianna P P Zhang

2012-02-01

Full Text Available Ebolaviruses cause hemorrhagic fever with up to 90% lethality and in fatal cases, are characterized by early suppression of the host innate immune system. One of the proteins likely responsible for this effect is VP24. VP24 is known to antagonize interferon signaling by binding host karyopherin α proteins, thereby preventing them from transporting the tyrosine-phosphorylated transcription factor STAT1 to the nucleus. Here, we report that VP24 binds STAT1 directly, suggesting that VP24 can suppress at least two distinct branches of the interferon pathway. Here, we also report the first crystal structures of VP24, derived from different species of ebolavirus that are pathogenic (Sudan and nonpathogenic to humans (Reston. These structures reveal that VP24 has a novel, pyramidal fold. A site on a particular face of the pyramid exhibits reduced solvent exchange when in complex with STAT1. This site is above two highly conserved pockets in VP24 that contain key residues previously implicated in virulence. These crystal structures and accompanying biochemical analysis map differences between pathogenic and nonpathogenic viruses, offer templates for drug design, and provide the three-dimensional framework necessary for biological dissection of the many functions of VP24 in the virus life cycle.

Purification of recombinant budgerigar fledgling disease virus VP1 capsid protein and its ability for in vitro capsid assembly

Science.gov (United States)

Rodgers, R. E.; Chang, D.; Cai, X.; Consigli, R. A.; Spooner, B. S. (Principal Investigator)

1994-01-01

A recombinant system for the major capsid VP1 protein of budgerigar fledgling disease virus has been established. The VP1 gene was inserted into a truncated form of the pFlag-1 vector and expressed in Escherichia coli. The budgerigar fledgling disease virus VP1 protein was purified to near homogeneity by immunoaffinity chromatography. Fractions containing highly purified VP1 were pooled and found to constitute 3.3% of the original E. coli-expressed VP1 protein. Electron microscopy revealed that the VP1 protein was isolated as pentameric capsomeres. Electron microscopy also revealed that capsid-like particles were formed in vitro from purified VP1 capsomeres with the addition of Ca2+ ions and the removal of chelating and reducing agents.

Scandinavian object shift, remnant VP-topicalisation, verb particles and causatives

DEFF Research Database (Denmark)

Engels, Eva; Vikner, Sten

2013-01-01

constructions in Danish and Swedish, namely particle verb constructions and causative constructions with Danish "lade" and Swedish "låta" ‘let’. It is shown how differences in the VP-internal object position give rise to mirror image sequences concerning Object Shift in connection with verb second (Vº......On the basis of an examination of remnant VP-topicalisation constructions, this paper argues for an order preservation analysis of Scandinavian Object Shift. Extending the empirical database, we account for the phenomena in an Optimality Theoretic framework. The paper focusses on two particular...

A novel recombinant pseudorabies virus expressing parvovirus VP2 gene: Immunogenicity and protective efficacy in swine

Directory of Open Access Journals (Sweden)

Chen Dishi

2011-06-01

Full Text Available Abstract Background Porcine parvovirus (PPV VP2 gene has been successfully expressed in many expression systems resulting in self-assembly of virus-like particles (VLPs with similar morphology to the native capsid. Here, a pseudorabies virus (PRV system was adopted to express the PPV VP2 gene. Methods A recombinant PRV SA215/VP2 was obtained by homologous recombination between the vector PRV viral DNA and a transfer plasmid. Then recombinant virus was purified with plaque purification, and its identity confirmed by PCR amplification, Western blot and indirect immunofluorescence (IFA analyses. Electronic microscopy of PRV SA215/VP2 confirmed self-assembly of both pseudorabies virus and VLPs from VP2 protein. Results Immunization of piglets with recombinant virus elicited PRV-specific and PPV-specific humoral immune responses and provided complete protection against a lethal dose of PRV challenges. Gilts immunized with recombinant viruses induced PPV-specific antibodies, and significantly reduced the mortality rate of (1 of 28 following virulent PPV challenge compared with the control (7 of 31. Furthermore, PPV virus DNA was not detected in the fetuses of recombinant virus immunized gilts. Conclusions In this study, a recombinant PRV SA215/VP2 virus expressing PPV VP2 protein was constructed using PRV SA215 vector. The safety, immunogenicity, and protective efficacy of the recombinant virus were demonstrated in piglets and primiparous gilts. This recombinant PRV SA215/VP2 represents a suitable candidate for the development of a bivalent vaccine against both PRV and PPV infection.

Alphavirus Replicon DNA Vectors Expressing Ebola GP and VP40 Antigens Induce Humoral and Cellular Immune Responses in Mice

Directory of Open Access Journals (Sweden)

Shoufeng Ren

2018-01-01

Full Text Available Ebola virus (EBOV causes severe hemorrhagic fevers in humans, and no approved therapeutics or vaccine is currently available. Glycoprotein (GP is the major protective antigen of EBOV, and can generate virus-like particles (VLPs by co-expression with matrix protein (VP40. In this study, we constructed a recombinant Alphavirus Semliki Forest virus (SFV replicon vector DREP to express EBOV GP and matrix viral protein (VP40. EBOV VLPs were successfully generated and achieved budding from 293 cells after co-transfection with DREP-based GP and VP40 vectors (DREP-GP+DREP-VP40. Vaccination of BALB/c mice with DREP-GP, DREP-VP40, or DREP-GP+DREP-VP40 vectors, followed by immediate electroporation resulted in a mixed IgG subclass production, which recognized EBOV GP and/or VP40 proteins. This vaccination regimen also led to the generation of both Th1 and Th2 cellular immune responses in mice. Notably, vaccination with DREP-GP and DREP-VP40, which produces both GP and VP40 antigens, induced a significantly higher level of anti-GP IgG2a antibody and increased IFN-γ secreting CD8+ T-cell responses relative to vaccination with DREP-GP or DREP-VP40 vector alone. Our study indicates that co-expression of GP and VP40 antigens based on the SFV replicon vector generates EBOV VLPs in vitro, and vaccination with recombinant DREP vectors containing GP and VP40 antigens induces Ebola antigen-specific humoral and cellular immune responses in mice. This novel approach provides a simple and efficient vaccine platform for Ebola disease prevention.

V.P. Naumenko Sociopolitical Activity (Second Half of XIX Century. – 1917 Year

Directory of Open Access Journals (Sweden)

Lilya R. Vegera

2014-08-01

Full Text Available The article investigates socio-political activity of V.P. Naumenko (1852-1919 – an outstanding representative of the Ukrainian intelligentsia of the late XIX – early XX century, politics, philologist, publisher, teacher. The scientist's career was developing under the influence of liberatory ideas during this period. The numerous works of eminent historians, who were contemporaries of V.P. Naumenko and researchers late XX – early XXI century, evidenced about it. Analysis of documentary materials proved that the impetus for an active social life of Vladimir Naumenko was family upbringing where service people attached great importance. The University of St. Vladimir in Kiev, where he was enrolled like a free listener in the 1868 had significant impact on the formation of personality and socio-political position of V.P. Naumenko. Scientist's activities in Kiev society where he held «right» liberal wing played an important role in his socio-political life. Vladimir Pavlovich along with other members of the «right» wing did not mind the ideas of socialism, but they treated them as a distant prospect. V.P. Naumenko saw the path to this ideal in peaceful reforms. V.P. Naumenko was also an active member of the liberal-democratic union – Ukrainian common nonpartisan democratic organization (UCNDO. Soon Vladimir Pavlovich stepped back from this participation, because in the early twentieth century the organization began to acquire a political character. Thus, the article analyzed V.P. Naumenko's socio-political activities and elucidate his role in the political process since the middle of the XIX century till the 1917 revolution.

VP-16 and alkylating agents activate a common metabolic pathway for suppression of DNA replication

International Nuclear Information System (INIS)

Das, S.K.; Berger, N.A.

1986-01-01

The cytotoxic effects of etoposide (VP-16) are mediated by topoisomerase II production of protein crosslinked DNA strand breaks. Previous studies have shown that alkylating agent induced DNA damage results in expansion of dTTP pools and reduction of dCTP pools and DNA replication. Studies were conducted with V79 cells to determine whether the metabolic consequences of VP-16 treatment were similar to those induced by alkylating agents. Treatment with 0.5μM VP-16 prolonged the doubling time of V79 cells from 12 to 18 hrs and caused cell volume to increase from 1.1 to 1.6 x 10 -12 l. 2mM caffeine completely blocked the volume increase and substantially prevented the prolongation of doubling time. 5μM VP-16 reduced the rate of [ 3 H]TdR incorporation by 70%, whereas in the presence of 2mM caffeine, VP-16 caused only a 10% decrease in the rate of [ 3 H]TdR incorporation. 4 hr treatment with 5.0μM VP-16 increased dTTP levels from 65 +/- 10 pmol/10 6 cells to 80 +/- 13 pmol/10 6 cells and caused dCTP level to decline from 113 +/- 23 pmol/10 6 cells to 92 +/- 17 pmol/10 6 cells. These results indicate that the metabolic consequences of VP-16 treatment are similar to alkylating agent treatment and that an increase in dTTP pools with a subsequent effect on ribonucleotide reductase may be a final common pathway by which many cytotoxic agents suppress DNA synthesis

Constraining the Dynamic Rupture Properties with Moment Tensor Derived Vp/Vs Ratios.

Science.gov (United States)

Smith-Boughner, L.; Baig, A. M.; Urbancic, T.; Viegas, G. F.

2014-12-01

The goal of hydraulic fracturing is to increase the permeability of rocks to extract hydrocarbons from "tight" formations. This process stimulates fluid-driven fractures which induce microseismic events. Successfully treating the formations, stimulating large volumes of the reservoir, depends on targeting parts of the formation with more "brittleness", a property which is frequently characterized from the mechanical properties of the rock. Typically, these properties are constrained using well-logs, vertical seismic profiles and 3-D seismic surveys. Such tools provide a static view of the reservoir on very large or very small scales. While lithology controls the average rock strength within a unit, the content (gas or fluid filled), the shape of the pore space and the concentration of micro-fractures alters the mechanical properties of the reservoir. Seismic moment tensor inversion of the events generated during these stimulations reveals that they are significantly non-double-couple, and are described by a tensile angle and a Poisson's ratio (or, equivalently, ratio of shear to compressional velocities, Vp/Vs) of the rock-fracture system. Following Vavryčuk (2011), the mechanical properties of the reservoir (i.e. Vp/Vs ratio) are estimated as the hydraulic fracture progresses from an extensive catalog of microseismic events spanning magnitudes of -1.5 to 0.8 in the Horn-River Basin, Canada. Studying several fracture stages in the reservoir reveals temporal and spatial variations in the rock strength within a unit as hydraulic fracturing proceeds. Initially, the estimated values of Vp/Vs are quite close to those determined from 3-D seismic surveys. As the stage progresses, previously fractured regions have lower Vp/Vs values. At the onset of maximum treating pressure, regions have anomalously high Vp/Vs values, which could reflect short-term local concentrations of high pore pressures or other interactions of the treatment with the formation. The relationship

Comparison of computerized digital and film-screen radiography: response to variation in imaging kVp

Energy Technology Data Exchange (ETDEWEB)

Broderick, N J; Long, B; Dreesen, R G; Cohen, M D; Cory, D A [Riley Hospital for Children, Indiana Univ. School of Medicine, Indianapolis, IN (United States). Dept. of Radiology; Katz, B P; Kalasinski, L A [Regenstreif Inst., Indiana Univ. School of Medicine, Indianapolis, IN (United States). Dept. of Medicine

1992-09-01

A controlled prospective study, in an animal model chosen to simulate portable neonatal radiography, was performed to compare the response of the Philips Computed Radiography (CR) system and conventional 200 speed film-screen (FS) to variation in imaging kVp. Acceptable images were obtained on the CR system over a very wide kVp range. In contrast the FS system produced acceptable images over a narrow kVp range. This ability suggests that the CR system should eliminate the need for repeat examinations in cases where a suboptimal kVp setting would have resulted in an unacceptable FS image. CR technology should therefore be ideally suited to portable radiography especially in situations where selection of correct exposure factors is difficult as in the neonatal nursery. (orig.).

Comparison of computerized digital and film-screen radiography: response to variation in imaging kVp

International Nuclear Information System (INIS)

Broderick, N.J.; Long, B.; Dreesen, R.G.; Cohen, M.D.; Cory, D.A.; Katz, B.P.; Kalasinski, L.A.

1992-01-01

A controlled prospective study, in an animal model chosen to simulate portable neonatal radiography, was performed to compare the response of the Philips Computed Radiography (CR) system and conventional 200 speed film-screen (FS) to variation in imaging kVp. Acceptable images were obtained on the CR system over a very wide kVp range. In contrast the FS system produced acceptable images over a narrow kVp range. This ability suggests that the CR system should eliminate the need for repeat examinations in cases where a suboptimal kVp setting would have resulted in an unacceptable FS image. CR technology should therefore be ideally suited to portable radiography especially in situations where selection of correct exposure factors is difficult as in the neonatal nursery. (orig.)

Regulation of the Osem gene by abscisic acid and the transcriptional activator VP1: analysis of cis-acting promoter elements required for regulation by abscisic acid and VP1.

Science.gov (United States)

Hattori, T; Terada, T; Hamasuna, S

1995-06-01

Osem, a rice gene homologous to the wheat Em gene, which encodes one of the late-embryogenesis abundant proteins was isolated. The gene was characterized with respect to control of transcription by abscisic acid (ABA) and the transcriptional activator VP1, which is involved in the ABA-regulated gene expression during late embryo-genesis. A fusion gene (Osem-GUS) consisting of the Osem promoter and the bacterial beta-glucuronidase (GUS) gene was constructed and tested in a transient expression system, using protoplasts derived from a suspension-cultured line of rice cells, for activation by ABA and by co-transfection with an expression vector (35S-Osvp1) for the rice VP1 (OSVP1) cDNA. The expression of Osem-GUS was strongly (40- to 150-fold) activated by externally applied ABA and by over-expression of (OS)VP1. The Osem promoter has three ACGTG-containing sequences, motif A, motif B and motif A', which resemble the abscisic acid-responsive element (ABRE) that was previously identified in the wheat Em and the rice Rab16. There is also a CATGCATG sequence, which is known as the Sph box and is shown to be essential for the regulation by VP1 of the maize anthocyanin regulatory gene C1. Focusing on these sequence elements, various mutant derivatives of the Osem promoter in the transient expression system were assayed. The analysis revealed that motif A functions not only as an ABRE but also as a sequence element required for the regulation by (OS)VP1.

Mutual antagonism between the Ebola virus VP35 protein and the RIG-I activator PACT determines infection outcome.

Science.gov (United States)

Luthra, Priya; Ramanan, Parameshwaran; Mire, Chad E; Weisend, Carla; Tsuda, Yoshimi; Yen, Benjamin; Liu, Gai; Leung, Daisy W; Geisbert, Thomas W; Ebihara, Hideki; Amarasinghe, Gaya K; Basler, Christopher F

2013-07-17

The cytoplasmic pattern recognition receptor RIG-I is activated by viral RNA and induces type I IFN responses to control viral replication. The cellular dsRNA binding protein PACT can also activate RIG-I. To counteract innate antiviral responses, some viruses, including Ebola virus (EBOV), encode proteins that antagonize RIG-I signaling. Here, we show that EBOV VP35 inhibits PACT-induced RIG-I ATPase activity in a dose-dependent manner. The interaction of PACT with RIG-I is disrupted by wild-type VP35, but not by VP35 mutants that are unable to bind PACT. In addition, PACT-VP35 interaction impairs the association between VP35 and the viral polymerase, thereby diminishing viral RNA synthesis and modulating EBOV replication. PACT-deficient cells are defective in IFN induction and are insensitive to VP35 function. These data support a model in which the VP35-PACT interaction is mutually antagonistic and plays a fundamental role in determining the outcome of EBOV infection. Copyright © 2013 Elsevier Inc. All rights reserved.

The decays Psub(c) -> VP in the group theoretical and quark diagrammatic approaches

International Nuclear Information System (INIS)

Tuan, S.F.; Xiaoyuan Li.

1983-08-01

Decays of charmed meson into one vector meson and one pseudoscalar meson Psub(c) -> VP in both the group theoretical and quark diagrammatic approaches are considered. A complete decay amplitude analysis is given. The present available experimental data can be accomodated if the contributions from exotic final states and exotic piece of weak Hamiltonian are also taken into account. (orig.)

Essential C-Terminal region of the baculovirus minor capsid protein VP80 binds DNA

NARCIS (Netherlands)

Marek, M.; Merten, O.W.; Francis-Devaraj, F.; Oers, van M.M.

2012-01-01

The essential Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) minor capsid protein VP80 has been recently shown to interact with the virus-triggered, nuclear F-actin cytoskeleton. A role for VP80 in virus morphogenesis has been proposed in the maturation of progeny nucleocapsids and

The ygaVP Genes of Escherichia coli Form a Tributyltin-Inducible Operon▿ †

OpenAIRE

Gueuné, Hervé; Durand, Marie-José; Thouand, Gérald; DuBow, Michael S.

2008-01-01

A tributyltin (TBT) luxAB transcriptional fusion in Escherichia coli revealed that a TBT-activated promoter is located upstream of two cotranscribed orphan genes, ygaV and ygaP. We demonstrate that transcription from the promoter upstream of ygaVP is constitutive in a ygaVP mutant, suggesting that YgaV is an autoregulated, TBT-inducible repressor.

Evolutionary and genetic analysis of the VP2 gene of canine parvovirus.

Science.gov (United States)

Li, Gairu; Ji, Senlin; Zhai, Xiaofeng; Zhang, Yuxiang; Liu, Jie; Zhu, Mengyan; Zhou, Jiyong; Su, Shuo

2017-07-17

Canine parvovirus (CPV) type 2 emerged in 1978 in the USA and quickly spread among dog populations all over the world with high morbidity. Although CPV is a DNA virus, its genomic substitution rate is similar to some RNA viruses. Therefore, it is important to trace the evolution of CPV to monitor the appearance of mutations that might affect vaccine effectiveness. Our analysis shows that the VP2 genes of CPV isolated from 1979 to 2016 are divided into six groups: GI, GII, GIII, GIV, GV, and GVI. Amino acid mutation analysis revealed several undiscovered important mutation sites: F267Y, Y324I, and T440A. Of note, the evolutionary rate of the CPV VP2 gene from Asia and Europe decreased. Codon usage analysis showed that the VP2 gene of CPV exhibits high bias with an ENC ranging from 34.93 to 36.7. Furthermore, we demonstrate that natural selection plays a major role compared to mutation pressure driving CPV evolution. There are few studies on the codon usage of CPV. Here, we comprehensively studied the genetic evolution, codon usage pattern, and evolutionary characterization of the VP2 gene of CPV. The novel findings revealing the evolutionary process of CPV will greatly serve future CPV research.

Characterization and specificity of the linear epitope of the enterovirus 71 VP2 protein

Directory of Open Access Journals (Sweden)

Kiener Tanja K

2012-02-01

Full Text Available Abstract Background Enterovirus 71 (EV71 has emerged as a major causative agent of hand, foot and mouth disease in the Asia-Pacific region over the last decade. Hand, foot and mouth disease can be caused by different etiological agents from the enterovirus family, mainly EV71 and coxsackieviruses, which are genetically closely related. Nevertheless, infection with EV71 may occasionally lead to high fever, neurologic complications and the emergence of a rapidly fatal syndrome of pulmonary edema associated with brainstem encephalitis. The rapid progression and high mortality of severe EV71 infection has highlighted the need for EV71-specific diagnostic and therapeutic tools. Monoclonal antibodies are urgently needed to specifically detect EV71 antigens from patient specimens early in the infection process. Furthermore, the elucidation of viral epitopes will contribute to the development of targeted therapeutics and vaccines. Results We have identified the monoclonal antibody 7C7 from a screen of hybridoma cells derived from mice immunized with the EV71-B5 strain. The linear epitope of 7C7 was mapped to amino acids 142-146 (EDSHP of the VP2 capsid protein and was characterized in detail. Mutational analysis of the epitope showed that the aspartic acid to asparagine mutation of the EV71 subgenogroup A (BrCr strain did not interfere with antibody recognition. In contrast, the serine to threonine mutation at position 144 of VP2, present in recently emerged EV71-C4 China strains, abolished antigenicity. Mice injected with this virus strain did not produce any antibodies against the VP2 protein. Immunofluorescence and Western blotting confirmed that 7C7 specifically recognized EV71 subgenogroups and did not cross-react to Coxsackieviruses 4, 6, 10, and 16. 7C7 was successfully used as a detection antibody in an antigen-capture ELISA assay. Conclusions Detailed mapping showed that the VP2 protein of Enterovirus 71 contains a single, linear, non

Human parvovirus B19 VP1u Protein as inflammatory mediators induces liver injury in naïve mice.

Science.gov (United States)

Hsu, Tsai-Ching; Chiu, Chun-Ching; Chang, Shun-Chih; Chan, Hsu-Chin; Shi, Ya-Fang; Chen, Tzy-Yen; Tzang, Bor-Show

2016-01-01

Human parvovirus B19 (B19V) is a human pathogen known to be associated with many non-erythroid diseases, including hepatitis. Although B19V VP1-unique region (B19-VP1u) has crucial roles in the pathogenesis of B19V infection, the influence of B19-VP1u proteins on hepatic injury is still obscure. This study investigated the effect and possible inflammatory signaling of B19-VP1u in livers from BALB/c mice that were subcutaneously inoculated with VP1u-expressing COS-7 cells. The in vivo effects of B19-VP1u were analyzed by using live animal imaging system (IVIS), Haematoxylin-Eosin staining, gel zymography, and immunoblotting after inoculation. Markedly hepatocyte disarray and lymphocyte infiltration, enhanced matrix metalloproteinase (MMP)-9 activity and increased phosphorylation of p38, ERK, IKK-α, IκB and NF-κB (p-p65) proteins were observed in livers from BALB/c mice receiving COS-7 cells expressing B19-VP1u as well as the significantly increased CRP, IL-1β and IL-6. Notably, IFN-γ and phosphorylated STAT1, but not STAT3, were also significantly increased in the livers of BALB/c mice that were subcutaneously inoculated with VP1u-expressing COS-7 cells. These findings revealed the effects of B19-VP1u on liver injury and suggested that B19-VP1u may have a role as mediators of inflammation in B19V infection.

Inhibition of rotavirus replication by a non-neutralizing, rotavirus VP6–specific IgA mAb

Science.gov (United States)

Feng, Ningguo; Lawton, Jeffrey A.; Gilbert, Joana; Kuklin, Nelly; Vo, Phuoc; Prasad, B.V. Venkataram; Greenberg, Harry B.

2002-01-01

Rotaviruses are the leading cause of severe diarrheal disease in young children. Intestinal mucosal IgA responses play a critical role in protective immunity against rotavirus reinfection. Rotaviruses consist of three concentric capsid layers surrounding a genome of 11 segments of double-stranded RNA. The outer layer proteins, VP4 and VP7, which are responsible for viral attachment and entry, are targets for protective neutralizing antibodies. However, IgA mAb’s directed against the intermediate capsid protein VP6, which do not neutralize the virus, have also been shown to protect mice from rotavirus infection and clear chronic infection in SCID mice. We investigated whether the anti-VP6 IgA (7D9) mAb could inhibit rotavirus replication inside epithelial cells and found that 7D9 acted at an early stage of infection to neutralize rotavirus following antibody lipofection. Using electron cryomicroscopy, we determined the three-dimensional structure of the virus-antibody complex. The attachment of 7D9 IgA to VP6 introduces a conformational change in the VP6 trimer, rendering the particle transcriptionally incompetent and preventing the elongation of initiated transcripts. Based on these observations, we suggest that anti-VP6 IgA antibodies confers protection in vivo by inhibiting viral transcription at the start of the intracellular phase of the viral replication cycle. PMID:11994409

The order-to-disorder transition behavior of PS-b-P2VP thin film system

Science.gov (United States)

Ahn, Hyungju; Ryu, Du

2013-03-01

We investigated the transition behavior such as the order-to-disorder transition (ODT) for symmetric poly(styrene)-block-poly(2-vinly pridine) (PS-b-P2VP) using SAXS and GISAXS for block copolymer bulks and films. The bulk transition temperature of PS-b-P2VP was significantly influenced by the interfacial interactions in thin films, leading to the different transition temperature. From these results, we will discuss about the interfacial interaction effects on the phase behaviors in bulks and thin films system of PS-b-P2VP.

Rotavirus capsid VP6 tubular and spherical nanostructures act as local adjuvants when co-delivered with norovirus VLPs.

Science.gov (United States)

Malm, M; Heinimäki, S; Vesikari, T; Blazevic, V

2017-09-01

A subunit protein vaccine candidate based on norovirus (NoV) virus-like particles (VLPs) and rotavirus (RV) VP6 protein against acute childhood gastroenteritis has been proposed recently. RV VP6 forms different oligomeric nanostructures, including tubes and spheres when expressed in vitro, which are highly immunogenic in different animal models. We have shown recently that recombinant VP6 nanotubes have an adjuvant effect on immunogenicity of NoV VLPs in mice. In this study, we investigated if the adjuvant effect is dependent upon a VP6 dose or different VP6 structural assemblies. In addition, local and systemic adjuvant effects as well as requirements for antigen co-delivery and co-localization were studied. The magnitude and functionality of NoV GII.4-specific antibodies and T cell responses were tested in mice immunized with GII.4 VLPs alone or different combinations of VLPs and VP6. A VP6 dose-dependent adjuvant effect on GII.4-specific antibody responses was observed. The adjuvant effect was found to be strictly dependent upon co-administration of NoV GII.4 VLPs and VP6 at the same anatomic site and at the same time. However, the adjuvant effect was not dependent on the types of oligomers used, as both nanotubes and nanospheres exerted adjuvant effect on GII.4-specific antibody generation and, for the first time, T cell immunity. These findings elucidate the mechanisms of VP6 adjuvant effect in vivo and support its use as an adjuvant in a combination NoV and RV vaccine. © 2017 The Authors. Clinical & Experimental Immunology published by John Wiley & Sons Ltd on behalf of British Society for Immunology.

Misperceptions about the risks of abortion in women presenting for abortion.

Science.gov (United States)

Wiebe, Ellen R; Littman, Lisa; Kaczorowski, Janusz; Moshier, Erin L

2014-03-01

Misinformation about the risks and sequelae of abortion is widespread. The purpose of this study was to examine whether women having an abortion who believe that there should be restrictions to abortion (i.e., that some other women should not be allowed to have an abortion) also believe this misinformation about the health risks associated with abortion. We carried out a cross-sectional survey of women presenting consecutively for an abortion at an urban abortion clinic in Vancouver, British Columbia, between February and September 2012. Of 1008 women presenting for abortion, 978 completed questionnaires (97% response rate), and 333 of these (34%) favoured abortion restrictions. More women who favoured restrictions believed that the health risk of an abortion was the same as or greater than the health risk of childbirth (84.2% vs. 65.6%, P abortion caused mental health problems (39.1% vs. 28.3%, P abortion caused infertility (41.7% vs. 21.9%, P abortion should not be restricted was found to be a significantly correlated with correct answers about health risks, mental health problems, and infertility. Misinformed beliefs about the risks of abortion are common among women having an abortion. Women presenting for abortion who favoured restrictions to abortion have more misperceptions about abortion risks than women who favour no restrictions.

Crystallization and preliminary X-ray diffraction analysis of the carbohydrate-recognizing domain (VP8*) of bovine rotavirus strain NCDV

International Nuclear Information System (INIS)

Yu, Xing; Guillon, Annabel; Szyczew, Alex J.; Kiefel, Milton J.; Coulson, Barbara S.; Itzstein, Mark von; Blanchard, Helen

2008-01-01

NCDV VP8* 64–224 was expressed in E. coli, purified and crystallized in the presence of a sialic acid derivative. X-ray diffraction data were obtained to a resolution of 2.0 Å and the crystallographic structure was determined by molecular replacement. The infectivity of rotavirus is dramatically enhanced by proteolytic cleavage of its outer layer VP4 spike protein into two functional domains, VP8* and VP5*. The carbohydrate-recognizing domain VP8* is proposed to bind sialic acid-containing host cell-surface glycans and this is followed by a series of subsequent virus–cell interactions. Live attenuated human and bovine rotavirus vaccine candidates for the prevention of gastroenteritis have been derived from bovine rotavirus strain NCDV. The NCDV VP8* 64–224 was overexpressed, purified to homogeneity and crystallized in the presence of an N-acetylneuraminic acid derivative. X-ray diffraction data were collected to a resolution of 2.0 Å and the crystallographic structure of NCDV VP8* 64–224 was determined by molecular replacement

A Medicago truncatula H+-pyrophosphatase gene, MtVP1, improves sucrose accumulation and anthocyanin biosynthesis in potato (Solanum tuberosum L.).

Science.gov (United States)

Wang, J W; Wang, H Q; Xiang, W W; Chai, T Y

2014-05-09

We recently cloned MtVP1, a type I vacuolar-type H(+)-translocating inorganic pyrophosphatase from Medicago truncatula. In the present study, we investigated the cellular location and the function of this H(+)-PPase in Arabidopsis and potato (Solanum tuberosum L.). An MtVP1::enhanced green fluorescent protein fusion was constructed, which localized to the plasma membrane of onion epidermal cells. Transgenic Arabidopsis thaliana overexpressing MtVP1 had more robust root systems and redder shoots than wild-type (WT) plants under conditions of cold stress. Furthermore, overexpression of MtVP1 in potato accelerated the formation and growth of vegetative organs. The tuber buds and stem base of transgenic potatoes became redder than those of WT plants, but flowering was delayed by approximately half a month. Interestingly, anthocyanin biosynthesis was promoted in transgenic Arabidopsis seedlings and potato tuber buds. The sucrose concentration of transgenic potato tubers and tuber buds was enhanced compared with that of WT plants. Furthermore, sucrose concentration in tubers was higher than that in tuber buds. Although there was no direct evidence to support Fuglsang's hypothetical model regarding the effects of H(+)-PPase on sucrose phloem loading, we speculated that sucrose concentration was increased in tuber buds owing to the increased concentration in tubers. Therefore, overexpressed MtVP1 enhanced sucrose accumulation of source organs, which might enhance sucrose transport to sink organs, thus affecting anthocyanin biosynthesis.

Joint 3-D tomographic imaging of Vp, Vs and Vp/Vs and hypocenter relocation at Sinabung volcano, Indonesia from November to December 2013

Science.gov (United States)

Nugraha, Andri Dian; Indrastuti, Novianti; Kusnandar, Ridwan; Gunawan, Hendra; McCausland, Wendy A.; Aulia, Atin Nur; Harlianti, Ulvienin

2018-01-01

We conducted travel time tomography using P- and S-wave arrival times of volcanic-tectonic (VT) events that occurred between November and December 2013 to determine the three-dimensional (3D) seismic velocity structure (Vp, Vs, and Vp/Vs) beneath Sinabung volcano, Indonesia in order to delineate geological subsurface structure and to enhance our understanding of the volcanism itself. This was a time period when phreatic explosions became phreatomagmatic and then magma migrated to the surface forming a summit lava dome. We used 4846 VT events with 16,138 P- and 16,138 S-wave arrival time phases recorded by 6 stations for the tomographic inversion. The relocated VTs collapse into three clusters at depths from the surface to sea level, from 2 to 4 km below sea level, and from 5 to 8.5 km below sea level. The tomographic inversion results show three prominent regions of high Vp/Vs (~ 1.8) beneath Sinabung volcano at depths consistent with the relocated earthquake clusters. We interpret these anomalies as intrusives associated with previous eruptions and possibly surrounding the magma conduit, which we cannot resolve with this study. One anomalous region might contain partial melt, at sea level and below the eventual eruption site at the summit. Our results are important for the interpretation of a conceptual model of the “plumbing system” of this hazardous volcano.

A novel life cycle modeling system for Ebola virus shows a genome length-dependent role of VP24 in virus infectivity.

Science.gov (United States)

Watt, Ari; Moukambi, Felicien; Banadyga, Logan; Groseth, Allison; Callison, Julie; Herwig, Astrid; Ebihara, Hideki; Feldmann, Heinz; Hoenen, Thomas

2014-09-01

Work with infectious Ebola viruses is restricted to biosafety level 4 (BSL4) laboratories, presenting a significant barrier for studying these viruses. Life cycle modeling systems, including minigenome systems and transcription- and replication-competent virus-like particle (trVLP) systems, allow modeling of the virus life cycle under BSL2 conditions; however, all current systems model only certain aspects of the virus life cycle, rely on plasmid-based viral protein expression, and have been used to model only single infectious cycles. We have developed a novel life cycle modeling system allowing continuous passaging of infectious trVLPs containing a tetracistronic minigenome that encodes a reporter and the viral proteins VP40, VP24, and GP1,2. This system is ideally suited for studying morphogenesis, budding, and entry, in addition to genome replication and transcription. Importantly, the specific infectivity of trVLPs in this system was ∼ 500-fold higher than that in previous systems. Using this system for functional studies of VP24, we showed that, contrary to previous reports, VP24 only very modestly inhibits genome replication and transcription when expressed in a regulated fashion, which we confirmed using infectious Ebola viruses. Interestingly, we also discovered a genome length-dependent effect of VP24 on particle infectivity, which was previously undetected due to the short length of monocistronic minigenomes and which is due at least partially to a previously unknown function of VP24 in RNA packaging. Based on our findings, we propose a model for the function of VP24 that reconciles all currently available data regarding the role of VP24 in nucleocapsid assembly as well as genome replication and transcription. Ebola viruses cause severe hemorrhagic fevers in humans, with no countermeasures currently being available, and must be studied in maximum-containment laboratories. Only a few of these laboratories exist worldwide, limiting our ability to study

Expression, purification, crystallization and preliminary X-ray diffraction analysis of the VP8* sialic acid-binding domain of porcine rotavirus strain OSU

International Nuclear Information System (INIS)

Zhang, Yang-De; Li, Hao; Liu, Hui; Pan, Yi-Feng

2007-01-01

Porcine rotavirus strain OSU VP8* domain has been expressed, purified and crystallized. X-ray diffraction data from different crystal forms of the VP8* domain have been collected to 2.65 and 2.2 Å resolution, respectively. The rotavirus outer capsid spike protein VP4 is utilized in the process of rotavirus attachment to and membrane penetration of host cells. VP4 is cleaved by trypsin into two domains: VP8* and VP5*. The VP8* domain is implicated in initial interaction with sialic acid-containing cell-surface carbohydrates and triggers subsequent virus invasion. The VP8* domain from porcine OSU rotavirus was cloned and expressed in Escherichia coli. Different crystal forms (orthorhombic P2 1 2 1 2 1 and tetragonal P4 1 2 1 2) were harvested from two distinct crystallization conditions. Diffraction data have been collected to 2.65 and 2.2 Å resolution and the VP8* 65–224 structure was determined by molecular replacement

Charmless two-body B(s)→VP decays in soft collinear effective theory

International Nuclear Information System (INIS)

Wang Wei; Wang Yuming; Yang Deshan; Lue Caidian

2008-01-01

We provide the analysis of charmless two-body B→VP decays under the framework of the soft collinear effective theory (SCET), where V(P) denotes a light vector (pseudoscalar) meson. Besides the leading power contributions, some power corrections (chiraly enhanced penguins) are also taken into account. Using the current available B→PP and B→VP experimental data on branching fractions and CP asymmetry variables, we find two kinds of solutions in χ 2 fit for the 16 nonperturbative inputs which are essential in the 87 B→PP and B→VP decay channels. Chiraly enhanced penguins can change several charming penguins sizably, since they share the same topology. However, most of the other nonperturbative inputs and predictions on branching ratios and CP asymmetries are not changed too much. With the two sets of inputs, we predict the branching fractions and CP asymmetries of other modes especially B s →VP decays. The agreements and differences with results in QCD factorization and perturbative QCD approach are analyzed. We also study the time-dependent CP asymmetries in channels with CP eigenstates in the final states and some other channels such as B 0 /B 0 →π ± ρ ± and B s 0 /B s 0 →K ± K* ± . In the perturbative QCD approach, the (S-P)(S+P) penguins in annihilation diagrams play an important role. Although they have the same topology with charming penguins in SCET, there are many differences between the two objects in weak phases, magnitudes, strong phases, and factorization properties.

Effect of reactive monomer on PS-b-P2VP film.

Science.gov (United States)

Kim, H J; Shin, D M

2014-08-01

Poly(styrene-b-2-vinyl pyridine) (PS-b-P2VP) lamellar film which is hydrophobic block-hydrophilic polyelectrolyte block polymer of 52 kg/mol-b-57 kg/mol and PS-b-P2VP film with reactive monomer (RM257) were prepared for photonic gel films. The lamellar stacks, which is alternating layer of hydrophilic and hydrophobic moiety of PS-b-P2VP, were obtained by exposing the spin coated film under chloroform vapor. The lamellar films were quaternized with 5 wt% of iodomethane diluted by n-hexane. We reported about the influence of reactive monomer on those photonic gel films. Added reactive monomer photonic gel film had higher absorbance than pure photonic gel films. As a result the photonic gel film with RM had more clear color. The lamellar films were swollen by DI water, ethanol (aq) and calcium carbonate solution. The band gaps of the lamellar films were drastically shifted to longer wavelength swollen by calcium carbonate solution. And the lamellar films were shifted to shorter wave length swollen by ethanol. So each lamellar film showed different color.

Characterization of nuclear localization and export signals of the major tegument protein VP8 of bovine herpesvirus-1

International Nuclear Information System (INIS)

Zheng Chunfu; Brownlie, Robert; Babiuk, Lorne A.; Hurk, Sylvia van Drunen Littel-van den

2004-01-01

Bovine herpesvirus-1 (BHV-1) VP8 is found in the nucleus immediately after infection. Transient expression of VP8 fused to yellow fluorescent protein (YFP) in COS-7 cells confirmed the nuclear localization of VP8 in the absence of other viral proteins. VP8 has four putative nuclear localization signals (NLS). Deletion of pat4 ( 51 RRPR 54 ) or pat7 ( 48 PRVRRPR 54 ) NLS2 abrogated nuclear accumulation, whereas deletion of 48 PRV 50 did not, so pat4 NLS2 is critical for nuclear localization of VP8. Furthermore, NLS1 ( 11 RRPRR 15 ), pat4 NLS2, and pat7 NLS2 were all capable of transporting the majority of YFP to the nucleus. Finally, a 12-amino-acid peptide with the sequence RRPRRPRVRRPR directed all of YFP into the nucleus, suggesting that reiteration of the RRPR motif makes the nuclear localization more efficient. Heterokaryon assays demonstrated that VP8 is also capable of shuttling between the nucleus and cytoplasm of the cell. Deletion mutant analysis revealed that this property is attributed to a leucine-rich nuclear export sequence (NES) consisting of amino acids 485 LSAYLTLFVAL 495 . This leucine-rich NES caused transport of YFP to the cytoplasm. These results demonstrate that VP8 shuttles between the nucleus and cytoplasm

Determinants of the VP1/2A junction cleavage by the 3C protease in foot-and-mouth disease virus-infected cells.

Science.gov (United States)

Kristensen, Thea; Normann, Preben; Gullberg, Maria; Fahnøe, Ulrik; Polacek, Charlotta; Rasmussen, Thomas Bruun; Belsham, Graham J

2017-03-01

The foot-and-mouth disease virus (FMDV) capsid precursor, P1-2A, is cleaved by FMDV 3C protease to yield VP0, VP3, VP1 and 2A. Cleavage of the VP1/2A junction is the slowest. Serotype O FMDVs with uncleaved VP1-2A (having a K210E substitution in VP1; at position P2 in cleavage site) have been described previously and acquired a second site substitution (VP1 E83K) during virus rescue. Furthermore, introduction of the VP1 E83K substitution alone generated a second site change at the VP1/2A junction (2A L2P, position P2' in cleavage site). These virus adaptations have now been analysed using next-generation sequencing to determine sub-consensus level changes in the virus; this revealed other variants within the E83K mutant virus population that changed residue VP1 K210. The construction of serotype A viruses with a blocked VP1/2A cleavage site (containing K210E) has now been achieved. A collection of alternative amino acid substitutions was made at this site, and the properties of the mutant viruses were determined. Only the presence of a positively charged residue at position P2 in the cleavage site permitted efficient cleavage of the VP1/2A junction, consistent with analyses of diverse FMDV genome sequences. Interestingly, in contrast to the serotype O virus results, no second site mutations occurred within the VP1 coding region of serotype A viruses with the blocked VP1/2A cleavage site. However, some of these viruses acquired changes in the 2C protein that is involved in enterovirus morphogenesis. These results have implications for the testing of potential antiviral agents targeting the FMDV 3C protease.

ALIX Rescues Budding of a Double PTAP/PPEY L-Domain Deletion Mutant of Ebola VP40: A Role for ALIX in Ebola Virus Egress.

Science.gov (United States)

Han, Ziying; Madara, Jonathan J; Liu, Yuliang; Liu, Wenbo; Ruthel, Gordon; Freedman, Bruce D; Harty, Ronald N

2015-10-01

Ebola (EBOV) is an enveloped, negative-sense RNA virus belonging to the family Filoviridae that causes hemorrhagic fever syndromes with high-mortality rates. To date, there are no licensed vaccines or therapeutics to control EBOV infection and prevent transmission. Consequently, the need to better understand the mechanisms that regulate virus transmission is critical to developing countermeasures. The EBOV VP40 matrix protein plays a central role in late stages of virion assembly and egress, and independent expression of VP40 leads to the production of virus-like particles (VLPs) by a mechanism that accurately mimics budding of live virus. VP40 late (L) budding domains mediate efficient virus-cell separation by recruiting host ESCRT and ESCRT-associated proteins to complete the membrane fission process. L-domains consist of core consensus amino acid motifs including PPxY, P(T/S)AP, and YPx(n)L/I, and EBOV VP40 contains overlapping PPxY and PTAP motifs whose interactions with Nedd4 and Tsg101, respectively, have been characterized extensively. Here, we present data demonstrating for the first time that EBOV VP40 possesses a third L-domain YPx(n)L/I consensus motif that interacts with the ESCRT-III protein Alix. We show that the YPx(n)L/I motif mapping to amino acids 18-26 of EBOV VP40 interacts with the Alix Bro1-V fragment, and that siRNA knockdown of endogenous Alix expression inhibits EBOV VP40 VLP egress. Furthermore, overexpression of Alix Bro1-V rescues VLP production of the budding deficient EBOV VP40 double PTAP/PPEY L-domain deletion mutant to wild-type levels. Together, these findings demonstrate that EBOV VP40 recruits host Alix via a YPx(n)L/I motif that can function as an alternative L-domain to promote virus egress. © The Author 2015. Published by Oxford University Press on behalf of the Infectious Diseases Society of America. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

[Effects of canine IL-2 and IL-7 genes on enhancing immunogenicity of canine parvovirus VP2 gene vaccine in mice].

Science.gov (United States)

Chen, Huihui; Zhong, Fei; Li, Xiujin; Wang, Lu; Sun, Yan; Neng, Changai; Zhang, Kao; Li, Wenyan; Wen, Jiexia

2012-11-04

To investigate the effects of canine interleukin-2 (cIL-2) and cIL-7 genes on enhancing the immunogenicity of canine parvovirus (CPV) VP2 DNA vaccine. The bicistronic vectors of cIL-2 and cIL-7 genes were constructed using the eukaryotic expression vector containing internal ribosome entry site (IRES). The cIL-2/ cIL-7 dicistronic vector plus previously constructed vectors, including CPV VP2 DNA vaccine vector, cIL-2 vector and cIL-7 vector, were used to co-immunize mice with different combinations, consisting of VP2 alone, VP2 + cIL-2, VP2 + cIL-7 and VP2 + cIL-2/cIL-7. The VP2-specific antibody levels in immunized mice were measured by ELISA at different time post-immunization. The proliferation indices and interferon-gamma expression were measured by lymphocyte proliferation assay and ELISA, respectively. The cIL-2/cIL-7 bicistronic vector was correct and could mediate cIL-2 and cIL-7 gene expression in eukaryotic cells. Immunization results revealed that the antibody titers and the neutralizing antibody levels of the mice co-immunized with VP2 + cIL-7/cIL-2 vectors were significantly higher than that with either VP2 + cIL-2 vectors or VP2 + cIL-7 vectors (P vaccine.

Detection of lipid-induced structural changes of the Marburg virus matrix protein VP40 using hydrogen/deuterium exchange-mass spectrometry.

Science.gov (United States)

Wijesinghe, Kaveesha J; Urata, Sarah; Bhattarai, Nisha; Kooijman, Edgar E; Gerstman, Bernard S; Chapagain, Prem P; Li, Sheng; Stahelin, Robert V

2017-04-14

Marburg virus (MARV) is a lipid-enveloped virus from the Filoviridae family containing a negative sense RNA genome. One of the seven MARV genes encodes the matrix protein VP40, which forms a matrix layer beneath the plasma membrane inner leaflet to facilitate budding from the host cell. MARV VP40 (mVP40) has been shown to be a dimeric peripheral protein with a broad and flat basic surface that can associate with anionic phospholipids such as phosphatidylserine. Although a number of mVP40 cationic residues have been shown to facilitate binding to membranes containing anionic lipids, much less is known on how mVP40 assembles to form the matrix layer following membrane binding. Here we have used hydrogen/deuterium exchange (HDX) mass spectrometry to determine the solvent accessibility of mVP40 residues in the absence and presence of phosphatidylserine and phosphatidylinositol 4,5-bisphosphate. HDX analysis demonstrates that two basic loops in the mVP40 C-terminal domain make important contributions to anionic membrane binding and also reveals a potential oligomerization interface in the C-terminal domain as well as a conserved oligomerization interface in the mVP40 N-terminal domain. Lipid binding assays confirm the role of the two basic patches elucidated with HD/X measurements, whereas molecular dynamics simulations and membrane insertion measurements complement these studies to demonstrate that mVP40 does not appreciably insert into the hydrocarbon region of anionic membranes in contrast to the matrix protein from Ebola virus. Taken together, we propose a model by which association of the mVP40 dimer with the anionic plasma membrane facilitates assembly of mVP40 oligomers. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

Expression, purification, crystallization and preliminary X-ray diffraction analysis of the VP8* sialic acid-binding domain of porcine rotavirus strain OSU

Energy Technology Data Exchange (ETDEWEB)

Zhang, Yang-De, E-mail: zhangyd1960@yahoo.com.cn; Li, Hao [National Hepatobiliary and Enteric Surgery Research Center of The Ministry of Health, Xiangya Hospital, Central South University, Hunan Province (China); Liu, Hui; Pan, Yi-Feng [Biochemistry Laboratory, Institution of Biomedical Engineering, Central South University, Hunan Province (China); National Hepatobiliary and Enteric Surgery Research Center of The Ministry of Health, Xiangya Hospital, Central South University, Hunan Province (China)

2007-02-01

Porcine rotavirus strain OSU VP8* domain has been expressed, purified and crystallized. X-ray diffraction data from different crystal forms of the VP8* domain have been collected to 2.65 and 2.2 Å resolution, respectively. The rotavirus outer capsid spike protein VP4 is utilized in the process of rotavirus attachment to and membrane penetration of host cells. VP4 is cleaved by trypsin into two domains: VP8* and VP5*. The VP8* domain is implicated in initial interaction with sialic acid-containing cell-surface carbohydrates and triggers subsequent virus invasion. The VP8* domain from porcine OSU rotavirus was cloned and expressed in Escherichia coli. Different crystal forms (orthorhombic P2{sub 1}2{sub 1}2{sub 1} and tetragonal P4{sub 1}2{sub 1}2) were harvested from two distinct crystallization conditions. Diffraction data have been collected to 2.65 and 2.2 Å resolution and the VP8*{sub 65–224} structure was determined by molecular replacement.

Oral Vaccination with the Porcine Rotavirus VP4 Outer Capsid Protein Expressed by Lactococcus lactis Induces Specific Antibody Production

Directory of Open Access Journals (Sweden)

Yi-jing Li

2010-01-01

Full Text Available The objective of this study to design a delivery system resistant to the gastrointestinal environment for oral vaccine against porcine rotavirus. Lactococcus lactis NZ9000 was transformed with segments of vP4 of the porcine rotavirus inserted into the pNZ8112 surface-expression vector, and a recombinant L. lactis expressing VP4 protein was constructed. An approximately 27 kDa VP4 protein was confirmed by SDS-PAGE , Western blot and immunostaining analysis. BALB/c mice were immunized orally with VP4-expression recombinant L. lactis and cellular, mucosal and systemic humoral immune responses were examined. Specific anti-VP4 secretory IgA and IgG were found in feces, ophthalmic and vaginal washes and in serum. The induced antibodies demonstrated neutralizing effects on porcine rotavirus infection on MA104 cells. Our findings suggest that oral immunization with VP4-expressing L. lactis induced both specific local and systemic humoral and cellular immune responses in mice.

PEO-b-P4VP/Yttrium Hydroxide Hybrid Nanotubes as Supporter for Catalyst Gold Nanoparticles

Science.gov (United States)

Yang, Qian; Chen, Dao-yong

2012-06-01

The adsorption of poly (ethylene oxide)-b-poly(4-vinylpyridine)(PEO-b-P4VP) micelles onto the surface of yttrium hydroxide nanotubes (YNTs) resulted in the hybrid nanotubes with a dense P4VP inner layer and a stretched PEO outer layer surrounding YNTs. The dense P4VP layer was further stabilized by the crosslinking using 1,4-dibromobutane as the crosslinker. Then, the crosslinked hybrid nanotubes (CHNTs) were used as a novel nano supporter for loading the catalyst gold nanoparticles (GNPs) within the crosslinked P4VP layer. The resultant GNPs/CHNTs (GNTs loaded on CHNTs) were applied to catalyze the reduction reaction of p-nitrophenol. The results indicate that this novel nano supporter has advantages such as good dispersity in the suspension, high capacity in loading GNPs (0.87 mmol/g), high catalytic activity of the loaded GNPs (12.9 μmol-1min-1), and good reusability of GNTs/CHNTs.

Abdominal CT: An intra-individual comparison between virtual monochromatic spectral and polychromatic 120-kVp images obtained during the same examination

Energy Technology Data Exchange (ETDEWEB)

Yamada, Yoshitake, E-mail: yamada@rad.med.keio.ac.jp [Department of Diagnostic Radiology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582 (Japan); Jinzaki, Masahiro, E-mail: jinzaki@rad.med.keio.ac.jp [Department of Diagnostic Radiology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582 (Japan); Hosokawa, Takahiro, E-mail: snowglobe@infoseek.jp [Department of Diagnostic Radiology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582 (Japan); Tanami, Yutaka, E-mail: tanami@rad.med.keio.ac.jp [Department of Diagnostic Radiology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582 (Japan); Abe, Takayuki, E-mail: tabe@z5.keio.jp [Center for Clinical Research, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582 (Japan); Kuribayashi, Sachio, E-mail: skuribay@med.keio.ac.jp [Department of Diagnostic Radiology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582 (Japan)

2014-10-15

Highlights: • We compared virtual monochromatic spectral (VMS) images with 120-kVp images. • VMS images are generated using accurate two-material beam-hardening correction. • Abdominal 70-keV VMS images provide better image quality than 120-kVp images. • Iterative reconstruction can further improve the image quality of VMS images. - Abstract: Objectives: To compare quantitative and subjective image quality between virtual monochromatic spectral (VMS) and conventional polychromatic 120-kVp imaging performed during the same abdominal computed tomography (CT) examination. Materials and methods: Our institutional review board approved this prospective study; each participant provided written informed consent. 51 patients underwent sequential fast kVp-switching dual-energy (80/140 kVp, volume CT dose index: 12.7 mGy) and single-energy (120-kVp, 12.7 mGy) abdominal enhanced CT over an 8 cm scan length with a random acquisition order and a 4.3-s interval. VMS images with filtered back projection (VMS-FBP) and adaptive statistical iterative reconstruction (so-called hybrid IR) (VMS-ASIR) (at 70 keV), as well as 120-kVp images with FBP (120-kVp-FBP) and ASIR (120-kVp-ASIR), were generated from dual-energy and single-energy CT data, respectively. The objective image noises, signal-to-noise ratios and contrast-to-noise ratios of the liver, kidney, pancreas, spleen, portal vein and aorta, and the lesion-to-liver and lesion-to-kidney contrast-to-noise ratios were measured. Two radiologists independently and blindly assessed the subjective image quality. The results were analyzed using the paired t-test, Wilcoxon signed rank sum test and mixed-effects model with Bonferroni correction. Results: VMS-ASIR images were superior to 120-kVp-FBP, 120-kVp-ASIR and VMS-FBP images for all the quantitative assessments and the subjective overall image quality (all P < 0.001), while VMS-FBP images were superior to 120-kVp-FBP and 120-kVp-ASIR images (all P < 0.004). Conclusions: VMS

Abdominal CT: An intra-individual comparison between virtual monochromatic spectral and polychromatic 120-kVp images obtained during the same examination

International Nuclear Information System (INIS)

Yamada, Yoshitake; Jinzaki, Masahiro; Hosokawa, Takahiro; Tanami, Yutaka; Abe, Takayuki; Kuribayashi, Sachio

2014-01-01

Highlights: • We compared virtual monochromatic spectral (VMS) images with 120-kVp images. • VMS images are generated using accurate two-material beam-hardening correction. • Abdominal 70-keV VMS images provide better image quality than 120-kVp images. • Iterative reconstruction can further improve the image quality of VMS images. - Abstract: Objectives: To compare quantitative and subjective image quality between virtual monochromatic spectral (VMS) and conventional polychromatic 120-kVp imaging performed during the same abdominal computed tomography (CT) examination. Materials and methods: Our institutional review board approved this prospective study; each participant provided written informed consent. 51 patients underwent sequential fast kVp-switching dual-energy (80/140 kVp, volume CT dose index: 12.7 mGy) and single-energy (120-kVp, 12.7 mGy) abdominal enhanced CT over an 8 cm scan length with a random acquisition order and a 4.3-s interval. VMS images with filtered back projection (VMS-FBP) and adaptive statistical iterative reconstruction (so-called hybrid IR) (VMS-ASIR) (at 70 keV), as well as 120-kVp images with FBP (120-kVp-FBP) and ASIR (120-kVp-ASIR), were generated from dual-energy and single-energy CT data, respectively. The objective image noises, signal-to-noise ratios and contrast-to-noise ratios of the liver, kidney, pancreas, spleen, portal vein and aorta, and the lesion-to-liver and lesion-to-kidney contrast-to-noise ratios were measured. Two radiologists independently and blindly assessed the subjective image quality. The results were analyzed using the paired t-test, Wilcoxon signed rank sum test and mixed-effects model with Bonferroni correction. Results: VMS-ASIR images were superior to 120-kVp-FBP, 120-kVp-ASIR and VMS-FBP images for all the quantitative assessments and the subjective overall image quality (all P < 0.001), while VMS-FBP images were superior to 120-kVp-FBP and 120-kVp-ASIR images (all P < 0.004). Conclusions: VMS

Unexpected detection of porcine rotavirus C strains carrying human origin VP6 gene.

Science.gov (United States)

Kattoor, Jobin Jose; Saurabh, Sharad; Malik, Yashpal Singh; Sircar, Shubhankar; Dhama, Kuldeep; Ghosh, Souvik; Bányai, Krisztián; Kobayashi, Nobumichi; Singh, Raj Kumar

2017-12-01

Rotavirus C (RVC), a known etiological agent of diarrheal outbreaks, mainly inflicts swine population globally with sporadic incidence in human, cattle, ferret, mink and dog. To demonstrate the presence of RVC in Indian swine population and characterization of its selected structural (VP6) and non-structural (NSP4 and NSP5) genes. A total of 108 diarrheic samples from different regions of India were used. Isolated RNA was loaded onto polyacrylamide gel to screen for the presence of RVs through the identification of specific electrophoretic genomic migration pattern. To characterize the RVC strains, VP6 gene and NSP4 and NSP5 genes were amplified, sequenced and analyzed. Based on VP6 gene specific diagnostic RT-PCR, the presence of RVC was confirmed in 12.0% (13/108) piglet fecal specimens. The nucleotide sequence analysis of VP6 gene, encoding inner capsid protein, from selected porcine RVC (PoRVC) strains revealed more than 93% homologies to human RVC strains (HuRVC) of Eurasian origin. These strains were distant from hitherto reported PoRVCs and clustered with HuRVCs, owning I2 genotype. However, the two non-structural genes, i.e. NSP4 and NSP5, of these strains were found to be of swine type, signifying a re-assortment event that has occurred in the Indian swine population. The findings indicate the presence of human-like RVC in Indian pigs and division of RVC clade with I2 genotype into further sub-clades. To the best of our knowledge, this appears to be the first report of RVC in Indian swine population. Incidence of human-like RVC VP6 gene in swine supports its subsequent zoonotic prospective.

Combination of VP3 and CD147-knockdown enhance apoptosis and tumor growth delay index in colorectal tumor allograft

International Nuclear Information System (INIS)

Ismail, Ruzila; Allaudin, Zeenathul Nazariah; Abdullah, Rasedee; Mohd Lila, Mohd-Azmi; Rahman, Nik-Mohd-Afizan Nik Abd.; Abdul Rahman, Sheikh-Omar

2016-01-01

Cancer therapies that kill cancer cells without affecting normal cells is the ultimate mode of treating cancers. The VP3, an avian virus-derived protein, can specifically initiate cell death through several signal transduction pathways leading to apoptosis. In cancer, chemoresistance and cell survivability implicate the cell surface protein, CD147. In this study, transfection of VP3 and silencing of CD147 genes was achieved through the treatment of tumors with pVIVO1-GFP/VP3 (VP3), psiRNA-CD147/2 (shCD147/2), and their combination of CT26 colon cancer cell-induced in mice. The effectiveness of tumor-treatment was ascertained by electrophoresis, TUNEL assay, and flow cytometry analysis. While histopathological and biochemical analysis were used as toxic side effect identification. The tumor growth delay index (TGDI) after treatment with VP3, shCD147/2, and their combination treatments increased by 1.3-, 1.2-, 2.0- and 2.3-fold respectively, over untreated control. The VP3-shCD147/2 combination treatment was more efficacious then either VP3 or shCD147/2 alone in the retardation of mouse CT26 colorectal cell tumor allograft. The antitumor effect of the combination treatment is the result of synergistic effects of VP3 and shCD147/2 on the tumor cells resulting in apoptosis. Thus, the study shows that combination of VP3 and shCD147/2 treatment can be developed into a potential approach for anticolorectal cancer treatment regimen. The online version of this article (doi:10.1186/s12885-016-2530-8) contains supplementary material, which is available to authorized users

Characterization of the RNA silencing suppression activity of the Ebola virus VP35 protein in plants and mammalian cells.

Science.gov (United States)

Zhu, Yali; Cherukuri, Nil Celebi; Jackel, Jamie N; Wu, Zetang; Crary, Monica; Buckley, Kenneth J; Bisaro, David M; Parris, Deborah S

2012-03-01

Ebola virus (EBOV) causes a lethal hemorrhagic fever for which there is no approved effective treatment or prevention strategy. EBOV VP35 is a virulence factor that blocks innate antiviral host responses, including the induction of and response to alpha/beta interferon. VP35 is also an RNA silencing suppressor (RSS). By inhibiting microRNA-directed silencing, mammalian virus RSSs have the capacity to alter the cellular environment to benefit replication. A reporter gene containing specific microRNA target sequences was used to demonstrate that prior expression of wild-type VP35 was able to block establishment of microRNA silencing in mammalian cells. In addition, wild-type VP35 C-terminal domain (CTD) protein fusions were shown to bind small interfering RNA (siRNA). Analysis of mutant proteins demonstrated that reporter activity in RSS assays did not correlate with their ability to antagonize double-stranded RNA (dsRNA)-activated protein kinase R (PKR) or bind siRNA. The results suggest that enhanced reporter activity in the presence of VP35 is a composite of nonspecific translational enhancement and silencing suppression. Moreover, most of the specific RSS activity in mammalian cells is RNA binding independent, consistent with VP35's proposed role in sequestering one or more silencing complex proteins. To examine RSS activity in a system without interferon, VP35 was tested in well-characterized plant silencing suppression assays. VP35 was shown to possess potent plant RSS activity, and the activities of mutant proteins correlated strongly, but not exclusively, with RNA binding ability. The results suggest the importance of VP35-protein interactions in blocking silencing in a system (mammalian) that cannot amplify dsRNA.

Controlled supramolecular assembly of micelle-like gold nanoparticles in PS-b-P2VP diblock copolymers via hydrogen bonding.

Science.gov (United States)

Jang, Se Gyu; Kramer, Edward J; Hawker, Craig J

2011-10-26

We report a facile strategy to synthesize amphiphilic gold (Au) nanoparticles functionalized with a multilayer, micelle-like structure consisting of a Au core, an inner hydroxylated polyisoprene (PIOH) layer, and an outer polystyrene shell (PS). Careful control of enthalpic interactions via a systematic variation of structural parameters, such as number of hydroxyl groups per ligand (N(OH)) and styrene repeating units (N(PS)) as well as areal chain density of ligands on the Au-core surface (Σ), enables precise control of the spatial distribution of these nanoparticles. This control was demonstrated in a lamellae-forming poly(styrene-b-2-vinylpyridine) (PS-b-P2VP) diblock copolymer matrix, where the favorable hydrogen-bonding interaction between hydroxyl groups in the PIOH inner shell and P2VP chains in the PS-b-P2VP diblock copolymer matrix, driving the nanoparticles to be segregated in P2VP domains, could be counter balanced by the enthalphic penalty of mixing of the PS outer brush with the P2VP domains. By varying N(OH), N(PS), and Σ, the nanoparticles could be positioned in the PS or P2VP domains or at the PS/P2VP interface. In addition, the effect of additives interfering with the hydrogen-bond formation between hydroxyl groups on Au nanoparticles and P2VP chains in a diblock copolymer matrix was investigated, and an interesting pea-pod-like segregation of Au nanoparticles in PS domains was observed.

Coupled adaptations affecting cleavage of the VP1/2A junction by 3C protease in foot-and-mouth disease virus infected cells

DEFF Research Database (Denmark)

Gullberg, Maria; Polacek, Charlotta; Belsham, Graham

The foot-and-mouth disease virus (FMDV) capsid protein precursor P1-2A is cleaved by the 3C protease to produce VP0, VP3, VP1 and 2A. It was shown previously that modification of a single amino acid residue (K210) within the VP1 protein, close to the VP1/2A cleavage site, inhibited cleavage......, introduction of the 2A L2P substitution alone, or with the VP1 K210E change, into this virus resulted in the production of viable viruses. Cells infected with viruses containing the VP1 K210E and/or the 2A L2P substitutions contained the uncleaved VP1-2A protein; the 2A L2P substitution rendered the VP1/2A...... of this junction and resulted in the production of “self-tagged” virus particles containing the 2A peptide. A second site substitution (E83K) within VP1 was also observed within the rescued virus (Gullberg et al., 2013). It is now shown that introduction of this E83K change alone into a serotype O virus resulted...

Inhibition of IRF-3 activation by VP35 is critical for the high level of virulence of ebola virus.

Science.gov (United States)

Hartman, Amy L; Bird, Brian H; Towner, Jonathan S; Antoniadou, Zoi-Anna; Zaki, Sherif R; Nichol, Stuart T

2008-03-01

Zaire ebolavirus causes a rapidly progressing hemorrhagic disease with high mortality. Identification of the viral virulence factors that contribute to the severity of disease induced by Ebola virus is critical for the design of therapeutics and vaccines against the disease. Given the rapidity of disease progression, virus interaction with the innate immune system early in the course of infection likely plays an important role in determining the outcome of the disease. The Ebola virus VP35 protein inhibits the activation of IRF-3, a critical transcription factor for the induction of early antiviral immunity. Previous studies revealed that a single amino acid change (R312A) in VP35 renders the protein unable to inhibit IRF-3 activation. A reverse-genetics-generated, mouse-adapted, recombinant Ebola virus that encodes the R312A mutation in VP35 was produced. We found that relative to the case for wild-type virus containing the authentic VP35 sequence, this single amino acid change in VP35 renders the virus completely attenuated in mice. Given that these viruses differ by only a single amino acid in the IRF-3 inhibitory domain of VP35, the level of alteration of virulence is remarkable and highlights the importance of VP35 for the pathogenesis of Ebola virus.

Interaction of monomeric Ebola VP40 protein with a plasma membrane: A coarse-grained molecular dynamics (CGMD) simulation study.

Science.gov (United States)

Mohamad Yusoff, Mohamad Ariff; Abdul Hamid, Azzmer Azzar; Mohammad Bunori, Noraslinda; Abd Halim, Khairul Bariyyah

2018-06-01

Ebola virus is a lipid-enveloped filamentous virus that affects human and non-human primates and consists of several types of protein: nucleoprotein, VP30, VP35, L protein, VP40, VP24, and transmembrane glycoprotein. Among the Ebola virus proteins, its matrix protein VP40 is abundantly expressed during infection and plays a number of critical roles in oligomerization, budding and egress from the host cell. VP40 exists predominantly as a monomer at the inner leaflet of the plasma membrane, and has been suggested to interact with negatively charged lipids such as phosphatidylinositol 4,5-bisphosphate (PIP 2 ) and phosphatidylserine (PS) via its cationic patch. The hydrophobic loop at the C-terminal domain has also been shown to be important in the interaction between the VP40 and the membrane. However, details of the molecular mechanisms underpinning their interactions are not fully understood. This study aimed at investigating the effects of mutation in the cationic patch and hydrophobic loop on the interaction between the VP40 monomer and the plasma membrane using coarse-grained molecular dynamics simulation (CGMD). Our simulations revealed that the interaction between VP40 and the plasma membrane is mediated by the cationic patch residues. This led to the clustering of PIP 2 around the protein in the inner leaflet as a result of interactions between some cationic residues including R52, K127, K221, K224, K225, K256, K270, K274, K275 and K279 and PIP 2 lipids via electrostatic interactions. Mutation of the cationic patch or hydrophobic loop amino acids caused the protein to bind at the inner leaflet of the plasma membrane in a different orientation, where no significant clustering of PIP 2 was observed around the mutated protein. This study provides basic understanding of the interaction of the VP40 monomer and its mutants with the plasma membrane. Copyright © 2018 Elsevier Inc. All rights reserved.

Decreased LRIG1 in Human Ovarian Cancer Cell SKOV3 Upregulates MRP-1 and Contributes to the Chemoresistance of VP16.

Science.gov (United States)

Yang, Hua; Yao, Jun; Yin, Jiangpin; Wei, Xuan

2016-05-01

The leucine-rich repeats and immunoglobulin-like domains (LRIG) are used as tumor suppressors in clinical applications. Although the LRIG has been identified to manipulate the cell proliferation via various oncogenic receptor tyrosine kinases in diverse cancers, its role in multidrug resistance needs to be further elucidated, especially in human ovarian cancer. We herein established that the etoposide (VP16)-resistant SKOV3 human ovarian cancer cell clones (SKOV3/VP16 cells) and mRNA expression of LRIG1 were significantly reduced by the treatment of VP16 in a concentration-dependent manner. Moreover, downregulated LRIG1 in SKOV3 could enhance the colony formation and resist the inhibition of proliferation by VP16, leading to the elevated expression of Bcl-2 and decreased apoptosis of SKOV3. Interestingly, our results uncovered that the multidrug resistance-associated protein 1 (MRP-1) was upregulated for the chemoresistance of VP16. To overcome the chemoresistance of SKOV3, SKOV3/VP16 was ectopically expressed of LRIG1. We found that the inhibition of VP16 on colony formation and proliferation was remarkably enhanced with increased apoptosis in SKOV3/VP16. Furthermore, the expression of MRP-1 and Bcl-2 was also inhibited, suggesting that the LRIG1could negatively control MRP-1 and the apoptosis to improve the sensitivity of VP16-related chemotherapy.

Rotavirus A strains obtained from children with acute gastroenteritis in Mozambique, 2012-2013: G and P genotypes and phylogenetic analysis of VP7 and partial VP4 genes.

Science.gov (United States)

João, Eva Dora; Strydom, Amy; O'Neill, Hester G; Cuamba, Assa; Cassocera, Marta; Acácio, Sozinho; Mandomando, Inácio; Motanyane, Lithabiso; Page, Nicola; de Deus, Nilsa

2018-01-01

In Mozambique rotavirus (RV) was shown to be the greatest cause of acute diarrhoea in infants from 0 to 11 months, and in 2015, national rotavirus vaccination was introduced. As with other developing countries, there is very limited active strain characterisation. Rotavirus positive clinical specimens, collected between 2012 and 2013, have now provided information on the genotypes circulating in southern Mozambique prior to vaccine introduction. Genotypes G2 (32.4%), G12 (28.0%), P[4] (41.4%) and P[6] (22.9%) (n = 157) strains were commonly detected with G2P[4] (42.3%) RVs being predominant, specifically during 2013. Phylogenetic evaluation of the VP7 and VP8* encoding genes showed, for the majority of the Mozambican strains, that they clustered with other African strains based on genotype. RVA/Human-wt/MOZ/0153/2013/G2P[4], RVA/Human-wt/MOZ/0308/2012/G2P[4] and RVA/Human-wt/MOZ/0288/2012/G12P[8] formed separate clusters from the other Mozambican strains with similar genotypes, suggesting possible reassortment. Amino acid substitutions in selected epitope regions also supported phylogenetic clustering. As expected, the VP7 and VP8* genes from the Mozambican strains differed from both the RotaTeq ® (SC2-9) G2P[5] and Rotarix ® (A41CB052A) G1P[8] genes. This study provides information on the genetic diversity of rotavirus strains prior to vaccine introduction and generates baseline data for future monitoring of any changes in rotavirus strains in response to vaccine pressure.

The Receptor-Binding Domain in the VP1u Region of Parvovirus B19.

Science.gov (United States)

Leisi, Remo; Di Tommaso, Chiarina; Kempf, Christoph; Ros, Carlos

2016-02-24

Parvovirus B19 (B19V) is known as the human pathogen causing the mild childhood disease erythema infectiosum. B19V shows an extraordinary narrow tissue tropism for erythroid progenitor cells in the bone marrow, which is determined by a highly restricted uptake. We have previously shown that the specific internalization is mediated by the interaction of the viral protein 1 unique region (VP1u) with a yet unknown cellular receptor. To locate the receptor-binding domain (RBD) within the VP1u, we analyzed the effect of truncations and mutations on the internalization capacity of the recombinant protein into UT7/Epo cells. Here we report that the N-terminal amino acids 5-80 of the VP1u are necessary and sufficient for cellular binding and internalization; thus, this N-terminal region represents the RBD required for B19V uptake. Using site-directed mutagenesis, we further identified a cluster of important amino acids playing a critical role in VP1u internalization. In silico predictions and experimental results suggest that the RBD is structured as a rigid fold of three α-helices. Finally, we found that dimerization of the VP1u leads to a considerably enhanced cellular binding and internalization. Taken together, we identified the RBD that mediates B19V uptake and mapped functional and structural motifs within this sequence. The findings reveal insights into the uptake process of B19V, which contribute to understand the pathogenesis of the infection and the neutralization of the virus by the immune system.

Removal of diethyl phthalate from aqueous phase using magnetic poly(EGDMA-VP) beads

Energy Technology Data Exchange (ETDEWEB)

Tuemay Oezer, Elif [Department of Chemistry, Uludag University, Bursa (Turkey); Osman, Bilgen, E-mail: bilgeno@uludag.edu.tr [Department of Chemistry, Uludag University, Bursa (Turkey); Kara, Ali; Besirli, Necati; Guecer, Seref [Department of Chemistry, Uludag University, Bursa (Turkey); Soezeri, Hueseyin [TUBITAK-UME, National Metrology Institute, PO Box 54 TR-41470, Gebze/Kocaeli (Turkey)

2012-08-30

Highlights: Black-Right-Pointing-Pointer Magnetic beads were prepared for removal of diethyl phthalate (DEP). Black-Right-Pointing-Pointer Total capacity of the beads was determined as 98.9 mg DEP per gram polymer. Black-Right-Pointing-Pointer Magnetic beads were regenerated easily and reused for DEP adsorption. Black-Right-Pointing-Pointer Adsorption isotherms, kinetics and thermodynamics were elucidated. - Abstract: The barium hexaferrite (BaFe{sub 12}O{sub 19}) containing magnetic poly(ethylene glycol dimethacrylate-vinyl pyridine), (mag-poly(EGDMA-VP)) beads (average diameter = 53-212 {mu}m) were synthesized and characterized. Their use as an adsorbent in the removal of diethyl phthalate (DEP) from an aqueous solution was investigated. The mag-poly(EGDMA-VP) beads were prepared by copolymerizing of 4-vinyl pyridine (VP) with ethylene glycol dimethacrylate (EGDMA). The mag-poly(EGDMA-VP) beads were characterized by N{sub 2} adsorption/desorption isotherms (BET), vibrating sample magnetometer (VSM), X-ray powder diffraction (XRD), elemental analysis, scanning electron microscope (SEM) and swelling studies. At a fixed solid/solution ratio, the various factors affecting the adsorption of DEP from aqueous solutions such as pH, initial concentration, contact time and temperature were analyzed. The maximum DEP adsorption capacity of the mag-poly(EGDMA-VP) beads was determined as 98.9 mg/g at pH 3.0, 25 Degree-Sign C. All the isotherm data can be fitted with both the Langmuir and the Dubinin-Radushkevich isotherm models. The pseudo first-order, pseudo-second-order, Ritch-second-order and intraparticle diffusion models were used to describe the adsorption kinetics. The thermodynamic parameters obtained indicated the exothermic nature of the adsorption. The DEP adsorption capacity did not change after 10 batch successive reactions, demonstrating the usefulness of the magnetic beads in applications.

Effects of Human Parvovirus B19 and Bocavirus VP1 Unique Region on Tight Junction of Human Airway Epithelial A549 Cells

Science.gov (United States)

Chiu, Chun-Ching; Shi, Ya-Fang; Yang, Jiann-Jou; Hsiao, Yuan-Chao; Tzang, Bor-Show; Hsu, Tsai-Ching

2014-01-01

As is widely recognized, human parvovirus B19 (B19) and human bocavirus (HBoV) are important human pathogens. Obviously, both VP1 unique region (VP1u) of B19 and HBoV exhibit the secreted phospholipase A2 (sPLA2)-like enzymatic activity and are recognized to participate in the pathogenesis of lower respiratory tract illnesses. However, exactly how, both VP1u from B19 and HBoV affect tight junction has seldom been addressed. Therefore, this study investigates how B19-VP1u and HBoV-VP1u may affect the tight junction of the airway epithelial A549 cells by examining phospholipase A2 activity and transepithelial electrical resistance (TEER) as well as performing immunoblotting analyses. Experimental results indicate that TEER is more significantly decreased in A549 cells by treatment with TNF-α (10 ng), two dosages of B19-VP1u and BoV-VP1u (400 ng and 4000 ng) or bee venom PLA2 (10 ng) than that of the control. Accordingly, more significantly increased claudin-1 and decreased occludin are detected in A549 cells by treatment with TNF-α or both dosages of HBoV-VP1u than that of the control. Additionally, more significantly decreased Na+/K+ ATPase is observed in A549 cells by treatment with TNF-α, high dosage of B19-VP1u or both dosages of BoV-VP1u than that of the control. Above findings suggest that HBoV-VP1u rather than B19 VP1u likely plays more important roles in the disruption of tight junction in the airway tract. Meanwhile, this discrepancy appears not to be associated with the secreted phospholipase A2 (sPLA2)-like enzymatic activity. PMID:25268969

Recombinant Marburg viruses containing mutations in the IID region of VP35 prevent inhibition of Host immune responses.

Science.gov (United States)

Albariño, César G; Wiggleton Guerrero, Lisa; Spengler, Jessica R; Uebelhoer, Luke S; Chakrabarti, Ayan K; Nichol, Stuart T; Towner, Jonathan S

2015-02-01

Previous in vitro studies have demonstrated that Ebola and Marburg virus (EBOV and MARV) VP35 antagonize the host cell immune response. Moreover, specific mutations in the IFN inhibitory domain (IID) of EBOV and MARV VP35 that abrogate their interaction with virus-derived dsRNA, lack the ability to inhibit the host immune response. To investigate the role of MARV VP35 in the context of infectious virus, we used our reverse genetics system to generate two recombinant MARVs carrying specific mutations in the IID region of VP35. Our data show that wild-type and mutant viruses grow to similar titers in interferon deficient cells, but exhibit attenuated growth in interferon-competent cells. Furthermore, in contrast to wild-type virus, both MARV mutants were unable to inhibit expression of various antiviral genes. The MARV VP35 mutants exhibit similar phenotypes to those previously described for EBOV, suggesting the existence of a shared immune-modulatory strategy between filoviruses. Published by Elsevier Inc.

VP-Nets : Efficient automatic localization of key brain structures in 3D fetal neurosonography.

Science.gov (United States)

Huang, Ruobing; Xie, Weidi; Alison Noble, J

2018-04-23

Three-dimensional (3D) fetal neurosonography is used clinically to detect cerebral abnormalities and to assess growth in the developing brain. However, manual identification of key brain structures in 3D ultrasound images requires expertise to perform and even then is tedious. Inspired by how sonographers view and interact with volumes during real-time clinical scanning, we propose an efficient automatic method to simultaneously localize multiple brain structures in 3D fetal neurosonography. The proposed View-based Projection Networks (VP-Nets), uses three view-based Convolutional Neural Networks (CNNs), to simplify 3D localizations by directly predicting 2D projections of the key structures onto three anatomical views. While designed for efficient use of data and GPU memory, the proposed VP-Nets allows for full-resolution 3D prediction. We investigated parameters that influence the performance of VP-Nets, e.g. depth and number of feature channels. Moreover, we demonstrate that the model can pinpoint the structure in 3D space by visualizing the trained VP-Nets, despite only 2D supervision being provided for a single stream during training. For comparison, we implemented two other baseline solutions based on Random Forest and 3D U-Nets. In the reported experiments, VP-Nets consistently outperformed other methods on localization. To test the importance of loss function, two identical models are trained with binary corss-entropy and dice coefficient loss respectively. Our best VP-Net model achieved prediction center deviation: 1.8 ± 1.4 mm, size difference: 1.9 ± 1.5 mm, and 3D Intersection Over Union (IOU): 63.2 ± 14.7% when compared to the ground truth. To make the whole pipeline intervention free, we also implement a skull-stripping tool using 3D CNN, which achieves high segmentation accuracy. As a result, the proposed processing pipeline takes a raw ultrasound brain image as input, and output a skull-stripped image with five detected key brain

Silver vanadium diphosphate Ag2VP2O8: Electrochemistry and characterization of reduced material providing mechanistic insights

International Nuclear Information System (INIS)

Takeuchi, Esther S.; Lee, Chia-Ying; Cheng, Po-Jen; Menard, Melissa C.; Marschilok, Amy C.; Takeuchi, Kenneth J.

2013-01-01

Silver vanadium phosphorous oxides (Ag w V x P y O z ) are notable battery cathode materials due to their high energy density and demonstrated ability to form in-situ Ag metal nanostructured electrically conductive networks within the cathode. While analogous silver vanadium diphosphate materials have been prepared, electrochemical evaluations of these diphosphate based materials have been limited. We report here the first electrochemical study of a silver vanadium diphosphate, Ag 2 VP 2 O 8 , where the structural differences associated with phosphorous oxides versus diphosphates profoundly affect the associated electrochemistry. Reminiscent of Ag 2 VO 2 PO 4 reduction, in-situ formation of silver metal nanoparticles was observed with reduction of Ag 2 VP 2 O 8 . However, counter to Ag 2 VO 2 PO 4 reduction, Ag 2 VP 2 O 8 demonstrates a significant decrease in conductivity upon continued electrochemical reduction. Structural analysis contrasting the crystallography of the parent Ag 2 VP 2 O 8 with that of the proposed Li 2 VP 2 O 8 reduction product is employed to gain insight into the observed electrochemical reduction behavior, where the structural rigidity associated with the diphosphate anion may be associated with the observed particle fracturing upon deep electrochemical reduction. Further, the diphosphate anion structure may be associated with the high thermal stability of the partially reduced Ag 2 VP 2 O 8 materials, which bodes well for enhanced safety of batteries incorporating this material. - Graphical abstract: Structure and galvanostatic intermittent titration-type test data for silver vanadium diphosphate, Ag 2 VP 2 O 8 . Highlights: ► First electrochemical study of a silver vanadium diphosphate, Ag 2 VP 2 O 8 . ► In-situ formation of Ag 0 nanoparticles was observed upon electrochemical reduction. ► Structural analysis used to provide insight of the electrochemical behavior

Expression of the VP2 protein of murine norovirus by a translation termination-reinitiation strategy.

Directory of Open Access Journals (Sweden)

Sawsan Napthine

2009-12-01

Full Text Available Expression of the minor virion structural protein VP2 of the calicivirus murine norovirus (MNV is believed to occur by the unusual mechanism of termination codon-dependent reinitiation of translation. In this process, following translation of an upstream open reading frame (ORF and termination at the stop codon, a proportion of 40S subunits remain associated with the mRNA and reinitiate at the AUG of a downstream ORF, which is typically in close proximity. Consistent with this, the VP2 start codon (AUG of MNV overlaps the stop codon of the upstream VP1 ORF (UAA in the pentanucleotide UAAUG.Here, we confirm that MNV VP2 expression is regulated by termination-reinitiation and define the mRNA sequence requirements. Efficient reintiation is dependent upon 43 nt of RNA immediately upstream of the UAAUG site. Chemical and enzymatic probing revealed that the RNA in this region is not highly structured and includes an essential stretch of bases complementary to 18S rRNA helix 26 (Motif 1. The relative position of Motif 1 with respect to the UAAUG site impacts upon the efficiency of the process. Termination-reinitiation in MNV was also found to be relatively insensitive to the initiation inhibitor edeine.The termination-reinitiation signal of MNV most closely resembles that of influenza BM2. Similar to other viruses that use this strategy, base-pairing between mRNA and rRNA is likely to play a role in tethering the 40S subunit to the mRNA following termination at the VP1 stop codon. Our data also indicate that accurate recognition of the VP2 ORF AUG is not a pre-requisite for efficient reinitiation of translation in this system.

Joint Inversion of Vp, Vs, and Resistivity at SAFOD

Science.gov (United States)

Bennington, N. L.; Zhang, H.; Thurber, C. H.; Bedrosian, P. A.

2010-12-01

Seismic and resistivity models at SAFOD have been derived from separate inversions that show significant spatial similarity between the main model features. Previous work [Zhang et al., 2009] used cluster analysis to make lithologic inferences from trends in the seismic and resistivity models. We have taken this one step further by developing a joint inversion scheme that uses the cross-gradient penalty function to achieve structurally similar Vp, Vs, and resistivity images that adequately fit the seismic and magnetotelluric MT data without forcing model similarity where none exists. The new inversion code, tomoDDMT, merges the seismic inversion code tomoDD [Zhang and Thurber, 2003] and the MT inversion code Occam2DMT [Constable et al., 1987; deGroot-Hedlin and Constable, 1990]. We are exploring the utility of the cross-gradients penalty function in improving models of fault-zone structure at SAFOD on the San Andreas Fault in the Parkfield, California area. Two different sets of end-member starting models are being tested. One set is the separately inverted Vp, Vs, and resistivity models. The other set consists of simple, geologically based block models developed from borehole information at the SAFOD drill site and a simplified version of features seen in geophysical models at Parkfield. For both starting models, our preliminary results indicate that the inversion produces a converging solution with resistivity, seismic, and cross-gradient misfits decreasing over successive iterations. We also compare the jointly inverted Vp, Vs, and resistivity models to borehole information from SAFOD to provide a "ground truth" comparison.

Magnetic Resonance Imaging Revealed Splenic Targeting of Canine Parvovirus Capsid Protein VP2

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Ma, Yufei; Wang, Haiming; Yan, Dan; Wei, Yanquan; Cao, Yuhua; Yi, Peiwei; Zhang, Hailu; Deng, Zongwu; Dai, Jianwu; Liu, Xiangtao; Luo, Jianxun; Zhang, Zhijun; Sun, Shiqi; Guo, Huichen

2016-03-01

Canine parvovirus (CPV) is a highly contagious infectious virus, whose infectious mechanism remains unclear because of acute gastroenteritis and the lack of an efficient tool to visualize the virus in real time during virology research. In this study, we developed an iron oxide nanoparticle supported by graphene quantum dots (GQD), namely, FeGQD. In this composite material, GQD acts as a stabilizer; thus, vacancies are retained on the surface for further physical adsorption of the CPV VP2 protein. The FeGQD@VP2 nanocomposite product showed largely enhanced colloidal stability in comparison with bare FeGQD, as well as negligible toxicity both in vitro and in vivo. The composite displayed high uptake into transferrin receptor (TfR) positive cells, which are distinguishable from FeGQD or TfR negative cells. In addition, the composite developed a significant accumulation in spleen rather than in liver, where bare FeGQD or most iron oxide nanoparticles gather. As these evident targeting abilities of FeGQD@VP2 strongly suggested, the biological activity of CPV VP2 was retained in our study, and its biological functions might correspond to CPV when the rare splenic targeting ability is considered. This approach can be applied to numerous other biomedical studies that require a simple yet efficient approach to track proteins in vivo while retaining biological function and may facilitate virus-related research.

The VP7 Outer Capsid Protein of Rotavirus Induces Polyclonal B-Cell Activation

Science.gov (United States)

Blutt, Sarah E.; Crawford, Sue E.; Warfield, Kelly L.; Lewis, Dorothy E.; Estes, Mary K.; Conner, Margaret E.

2004-01-01

The early response to a homologous rotavirus infection in mice includes a T-cell-independent increase in the number of activated B lymphocytes in the Peyer's patches. The mechanism of this activation has not been previously determined. Since rotavirus has a repetitively arranged triple-layered capsid and repetitively arranged antigens can induce activation of B cells, one or more of the capsid proteins could be responsible for the initial activation of B cells during infection. To address this question, we assessed the ability of rotavirus and virus-like particles to induce B-cell activation in vivo and in vitro. Using infectious rotavirus, inactivated rotavirus, noninfectious but replication-competent virus, and virus-like particles, we determined that neither infectivity nor RNA was necessary for B-cell activation but the presence of the rotavirus outer capsid protein, VP7, was sufficient for murine B-cell activation. Preincubation of the virus with neutralizing VP7 antibodies inhibited B-cell activation. Polymyxin B treatment and boiling of the virus preparation were performed, which ruled out possible lipopolysaccharide contamination as the source of activation and confirmed that the structural conformation of VP7 is important for B-cell activation. These findings indicate that the structure and conformation of the outer capsid protein, VP7, initiate intestinal B-cell activation during rotavirus infection. PMID:15194774

Perinatal outcomes in pregnant women presenting with preterm ...

African Journals Online (AJOL)

Perinatal outcomes in pregnant women presenting with preterm premature rupture of membranes at a ... Journal Home > Vol 23, No 2 (2017) > ... in sepsis rates, the need for ventilation and the duration of hospital stay between the two groups.

The Novel Gene VpPR4-1 from Vitis pseudoreticulata Increases Powdery Mildew Resistance in Transgenic Vitis vinifera L.

Science.gov (United States)

Dai, Lingmin; Wang, Dan; Xie, Xiaoqing; Zhang, Chaohong; Wang, Xiping; Xu, Yan; Wang, Yuejin; Zhang, Jianxia

2016-01-01

Pathogenesis-related proteins (PRs) can lead to increased resistance of the whole plant to pathogen attack. Here, we isolate and characterize a PR-4 protein (VpPR4-1) from a wild Chinese grape Vitis pseudoreticulata which shows greatly elevated transcription following powdery mildew infection. Its expression profiles under a number of abiotic stresses were also investigated. Powdery mildew, salicylic acid, and jasmonic acid methyl ester significantly increased the VpPR4-1 induction while NaCl and heat treatments just slightly induced VpPR4-1 expression. Abscisic acid and cold treatment slightly affected the expression level of VpPR4-1. The VpPR4-1 gene was overexpressed in 30 regenerated V. vinifera cv. Red Globe via Agrobacterium tumefaciens-mediated transformation and verified by the Western blot. The 26 transgenic grapevines exhibited higher expression levels of PR-4 protein content than wild-type vines and six of them were inoculated with powdery mildew which showed that the growth of powdery mildew was repressed. The powdery mildew-resistance of Red Globe transformed with VpPR4-1 was enhanced inoculated with powdery mildew. Moreover, other powdery mildew resistant genes were associated with feedback regulation since VpPR4-1 is in abundance. This study demonstrates that PR-4 protein in grapes plays a vital role in defense against powdery mildew invasion.

Image quality and radiation dose of lower extremity CT angiography at 70 kVp on an integrated circuit detector dual-source computed tomography.

Science.gov (United States)

Qi, Li; Zhao, Yan'E; Zhou, Chang Sheng; Spearman, James V; Renker, Matthias; Schoepf, U Joseph; Zhang, Long Jiang; Lu, Guang Ming

2015-06-01

Despite the well-established requirement for radiation dose reduction there are few studies examining the potential for lower extremity CT angiography (CTA) at 70 kVp. To compare the image quality and radiation dose of lower extremity CTA at 70 kVp using a dual-source CT system with an integrated circuit detector to similar studies at 120 kVp. A total of 62 patients underwent lower extremity CTA. Thirty-one patients were examined at 70 kVp using a second generation dual-source CT with an integrated circuit detector (70 kVp group) and 31 patients were evaluated at 120 kVp using a first generation dual-source CT (120 kVp group). The attenuation and image noise were measured and signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) were calculated. Two radiologists assessed image quality. Radiation dose was compared. The mean attenuation of the 70 kVp group was higher than the 120 kVp group (575 ± 149 Hounsfield units [HU] vs. 258 ± 38 HU, respectively, P < 0.001) as was SNR (44.0 ± 22.0 vs 32.7 ± 13.3, respectively, P = 0.017), CNR (39.7 ± 20.6 vs 26.6 ± 11.7, respectively, P = 0.003) and the mean image quality score (3.7 ± 0.1 vs. 3.2 ± 0.3, respectively, P < 0.001). The inter-observer agreement was good for the 70 kVp group and moderate for the 120 kVp group. The dose-length product was lower in the 70 kVp group (264.5 ± 63.1 mGy × cm vs. 412.4 ± 81.5 mGy × cm, P < 0.001). Lower extremity CTA at 70 kVp allows for lower radiation dose with higher SNR, CNR, and image quality when compared with standard 120 kVp. © The Foundation Acta Radiologica 2014 Reprints and permissions: sagepub.co.uk/journalsPermissions.nav.

Radiation dose reduction using 100-kVp and a sinogram-affirmed iterative reconstruction algorithm in adolescent head CT: Impact on grey-white matter contrast and image noise.

Science.gov (United States)

Nagayama, Yasunori; Nakaura, Takeshi; Tsuji, Akinori; Urata, Joji; Furusawa, Mitsuhiro; Yuki, Hideaki; Hirarta, Kenichiro; Kidoh, Masafumi; Oda, Seitaro; Utsunomiya, Daisuke; Yamashita, Yasuyuki

2017-07-01

To retrospectively evaluate the image quality and radiation dose of 100-kVp scans with sinogram-affirmed iterative reconstruction (IR) for unenhanced head CT in adolescents. Sixty-nine patients aged 12-17 years underwent head CT under 120- (n = 34) or 100-kVp (n = 35) protocols. The 120-kVp images were reconstructed with filtered back-projection (FBP), 100-kVp images with FBP (100-kVp-F) and sinogram-affirmed IR (100-kVp-S). We compared the effective dose (ED), grey-white matter (GM-WM) contrast, image noise, and contrast-to-noise ratio (CNR) between protocols in supratentorial (ST) and posterior fossa (PS). We also assessed GM-WM contrast, image noise, sharpness, artifacts, and overall image quality on a four-point scale. ED was 46% lower with 100- than 120-kVp (p < 0.001). GM-WM contrast was higher, and image noise was lower, on 100-kVp-S than 120-kVp at ST (p < 0.001). CNR of 100-kVp-S was higher than of 120-kVp (p < 0.001). GM-WM contrast of 100-kVp-S was subjectively rated as better than of 120-kVp (p < 0.001). There were no significant differences in the other criteria between 100-kVp-S and 120-kVp (p = 0.072-0.966). The 100-kVp with sinogram-affirmed IR facilitated dramatic radiation reduction and better GM-WM contrast without increasing image noise in adolescent head CT. • 100-kVp head CT provides 46% radiation dose reduction compared with 120-kVp. • 100-kVp scanning improves subjective and objective GM-WM contrast. • Sinogram-affirmed IR decreases head CT image noise, especially in supratentorial region. • 100-kVp protocol with sinogram-affirmed IR is suited for adolescent head CT.

Sequence adaptations affecting cleavage of the VP1/2A junction by the 3C protease in foot-and-mouth disease virus-infected cells

DEFF Research Database (Denmark)

Gullberg, Maria; Polacek, Charlotta; Belsham, Graham

2014-01-01

The foot-and-mouth disease virus (FMDV) capsid protein precursor P1-2A is cleaved by the virus-encoded 3C protease to VP0, VP3, VP1 and 2A. It was shown previously that modification of a single amino acid residue (K210E) within the VP1 protein and close to the VP1/2A cleavage site, inhibited...... cleavage of this junction and produced 'self-tagged' virus particles. A second site substitution (E83K) within VP1 was also observed within the rescued virus [Gullberg et al. (2013). J Virol 87: , 11591-11603]. It was shown here that introduction of this E83K change alone into a serotype O virus resulted...... in the rapid accumulation of a second site substitution within the 2A sequence (L2P), which also blocked VP1/2A cleavage. This suggests a linkage between the E83K change in VP1 and cleavage of the VP1/2A junction. Cells infected with viruses containing the VP1 K210E or the 2A L2P substitutions contained...

Silencing the alarms: innate immune antagonism by rotavirus NSP1 and VP3

Science.gov (United States)

Morelli, Marco; Ogden, Kristen M.; Patton, John T.

2016-01-01

The innate immune response involves a broad array of pathogen sensors that stimulate the production of interferons (IFN) to induce an antiviral state. Rotavirus, a significant cause of childhood gastroenteritis and a member of the Reoviridae family of segmented, double-stranded RNA viruses, encodes at least two direct antagonists of host innate immunity: NSP1 and VP3. NSP1, a putative E3 ubiquitin ligase, mediates the degradation of cellular factors involved in both IFN induction and downstream signaling. VP3, the viral capping enzyme, utilizes a 2H-phosphodiesterase domain to prevent activation of the cellular oligoadenylate synthase (OAS)-RNase L pathway. Computational, molecular, and biochemical studies have provided key insights into the structural and mechanistic basis of innate immune antagonism by NSP1 and VP3 of group A rotaviruses (RVA). Future studies with non-RVA isolates will be essential to understand how other RV species evade host innate immune responses. PMID:25724417

Comparative assessment of H.265/MPEG-HEVC, VP9, and H.264/MPEG-AVC encoders for low-delay video applications

Science.gov (United States)

Grois, Dan; Marpe, Detlev; Nguyen, Tung; Hadar, Ofer

2014-09-01

The popularity of low-delay video applications dramatically increased over the last years due to a rising demand for realtime video content (such as video conferencing or video surveillance), and also due to the increasing availability of relatively inexpensive heterogeneous devices (such as smartphones and tablets). To this end, this work presents a comparative assessment of the two latest video coding standards: H.265/MPEG-HEVC (High-Efficiency Video Coding), H.264/MPEG-AVC (Advanced Video Coding), and also of the VP9 proprietary video coding scheme. For evaluating H.264/MPEG-AVC, an open-source x264 encoder was selected, which has a multi-pass encoding mode, similarly to VP9. According to experimental results, which were obtained by using similar low-delay configurations for all three examined representative encoders, it was observed that H.265/MPEG-HEVC provides significant average bit-rate savings of 32.5%, and 40.8%, relative to VP9 and x264 for the 1-pass encoding, and average bit-rate savings of 32.6%, and 42.2% for the 2-pass encoding, respectively. On the other hand, compared to the x264 encoder, typical low-delay encoding times of the VP9 encoder, are about 2,000 times higher for the 1-pass encoding, and are about 400 times higher for the 2-pass encoding.

Integrated Computational Approach for Virtual Hit Identification against Ebola Viral Proteins VP35 and VP40

Directory of Open Access Journals (Sweden)

Muhammad Usman Mirza

2016-10-01

Full Text Available The Ebola virus (EBOV has been recognised for nearly 40 years, with the most recent EBOV outbreak being in West Africa, where it created a humanitarian crisis. Mortalities reported up to 30 March 2016 totalled 11,307. However, up until now, EBOV drugs have been far from achieving regulatory (FDA approval. It is therefore essential to identify parent compounds that have the potential to be developed into effective drugs. Studies on Ebola viral proteins have shown that some can elicit an immunological response in mice, and these are now considered essential components of a vaccine designed to protect against Ebola haemorrhagic fever. The current study focuses on chemoinformatic approaches to identify virtual hits against Ebola viral proteins (VP35 and VP40, including protein binding site prediction, drug-likeness, pharmacokinetic and pharmacodynamic properties, metabolic site prediction, and molecular docking. Retrospective validation was performed using a database of non-active compounds, and early enrichment of EBOV actives at different false positive rates was calculated. Homology modelling and subsequent superimposition of binding site residues on other strains of EBOV were carried out to check residual conformations, and hence to confirm the efficacy of potential compounds. As a mechanism for artefactual inhibition of proteins through non-specific compounds, virtual hits were assessed for their aggregator potential compared with previously reported aggregators. These systematic studies have indicated that a few compounds may be effective inhibitors of EBOV replication and therefore might have the potential to be developed as anti-EBOV drugs after subsequent testing and validation in experiments in vivo.

Integrated Computational Approach for Virtual Hit Identification against Ebola Viral Proteins VP35 and VP40.

Science.gov (United States)

Mirza, Muhammad Usman; Ikram, Nazia

2016-10-26

The Ebola virus (EBOV) has been recognised for nearly 40 years, with the most recent EBOV outbreak being in West Africa, where it created a humanitarian crisis. Mortalities reported up to 30 March 2016 totalled 11,307. However, up until now, EBOV drugs have been far from achieving regulatory (FDA) approval. It is therefore essential to identify parent compounds that have the potential to be developed into effective drugs. Studies on Ebola viral proteins have shown that some can elicit an immunological response in mice, and these are now considered essential components of a vaccine designed to protect against Ebola haemorrhagic fever. The current study focuses on chemoinformatic approaches to identify virtual hits against Ebola viral proteins (VP35 and VP40), including protein binding site prediction, drug-likeness, pharmacokinetic and pharmacodynamic properties, metabolic site prediction, and molecular docking. Retrospective validation was performed using a database of non-active compounds, and early enrichment of EBOV actives at different false positive rates was calculated. Homology modelling and subsequent superimposition of binding site residues on other strains of EBOV were carried out to check residual conformations, and hence to confirm the efficacy of potential compounds. As a mechanism for artefactual inhibition of proteins through non-specific compounds, virtual hits were assessed for their aggregator potential compared with previously reported aggregators. These systematic studies have indicated that a few compounds may be effective inhibitors of EBOV replication and therefore might have the potential to be developed as anti-EBOV drugs after subsequent testing and validation in experiments in vivo.

VP1u phospholipase activity is critical for infectivity of full-length parvovirus B19 genomic clones.

Science.gov (United States)

Filippone, Claudia; Zhi, Ning; Wong, Susan; Lu, Jun; Kajigaya, Sachiko; Gallinella, Giorgio; Kakkola, Laura; Söderlund-Venermo, Maria; Young, Neal S; Brown, Kevin E

2008-05-10

Three full-length genomic clones (pB19-M20, pB19-FL and pB19-HG1) of parvovirus B19 were produced in different laboratories. pB19-M20 was shown to produce infectious virus. To determine the differences in infectivity, all three plasmids were tested by transfection and infection assays. All three clones were similar in viral DNA replication, RNA transcription, and viral capsid protein production. However, only pB19-M20 and pB19-HG1 produced infectious virus. Comparison of viral sequences showed no significant differences in ITR or NS regions. In the capsid region, there was a nucleotide sequence difference conferring an amino acid substitution (E176K) in the phospholipase A2-like motif of the VP1-unique (VP1u) region. The recombinant VP1u with the E176K mutation had no catalytic activity as compared with the wild-type. When this mutation was introduced into pB19-M20, infectivity was significantly attenuated, confirming the critical role of this motif. Investigation of the original serum from which pB19-FL was cloned confirmed that the phospholipase mutation was present in the native B19 virus.

VP1u phospholipase activity is critical for infectivity of full-length parvovirus B19 genomic clones✰

Science.gov (United States)

Filippone, Claudia; Zhi, Ning; Wong, Susan; Lu, Jun; Kajigaya, Sachiko; Gallinella, Giorgio; Kakkola, Laura; Venermo, Maria S Söderlund; Young, Neal S.; Brown, Kevin E.

2008-01-01

Three full-length genomic clones (pB19-M20, pB19-FL and pB19-HG1) of parvovirus B19 were produced in different laboratories. pB19-M20 was shown to produce infectious virus. To determine the differences in infectivity, all three plasmids were tested by transfection and infection assays. All three clones were similar in viral DNA replication, RNA transcription, and viral capsid protein production. However, only pB19-M20 and pB19-HG1 produced infectious virus. Comparison of viral sequences showed no significant differences in ITR or NS regions. In the capsid region, there was a nucleotide sequence difference conferring an amino acid substitution (E176K) in the phospholipase A2-like motif of the VP1-unique (VP1u) region. The recombinant VP1u with the E176K mutation had no catalytic activity as compared with the wild-type. When this mutation was introduced into pB19-M20, infectivity was significantly attenuated, confirming the critical role of this motif. Investigation of the original serum from which pB19-FL was cloned confirmed that the phospholipase mutation was present in the native B19 virus. PMID:18252260

Combination of targeting gene-viro therapy with recombinant Fowl-pox viruses with HN and VP3 genes on mouse osteosarcoma.

Science.gov (United States)

Zhang, Z-Y; Wang, L-Q; Fu, C-F; Li, X; Cui, Z-L; Zhang, J-Y; Xue, S-H; Sun, N; Xu, F

2013-03-01

Osteosarcoma is an aggressive cancerous neoplasm arising from primitive transformed cells of mesenchymal origin that exhibit osteoblastic differentiation and produce malignant osteoid. With the rapid development of tumor molecular biology, gene and viral therapy, a highly promising strategy for the treatment, has shown some therapeutic effects. To study the strategy of cooperative cancer gene therapy, previously, we explored the antitumor effects of recombinant Fowl-pox viruses (FPVs) with both HN (hemagglutinin-neuramidinase) and VP3 genes on mouse osteosarcoma. We constructed vFV-HN, vFV-VP3 and vFV-HN-VP3 inserting CAV VP3 gene, NDV HN gene into fowlpox virus. S180 osteosarcoma were transfected with Recombinant Fowl-pox viruses (FPVs). These cell lines stably expressing tagged proteins were selected by culturing in medium containing puromycin (2 µg/ml) and confirmed by immunoblotting and immunostaining. S180 osteosarcoma model with BALB/c mice and nude mice were established and the vFPV viruses as control, vFV-HN, vFV-VP3, vFV-HN-VP3 were injected into the tumor directly. The rate of tumor growth, tumor suppression and the sialic acid levels in serum were examined and the tumor tissues were analyzed by the method of immunohistochemistry. Flow cytometric analysis was performed using a FACSCalibur flow cytometer. A total of 100,000 events were analyzed for each sample and the experiment was repeated at least twice. Our data indicated that vFV-HN, vFV-VP3 and vFV-HN-VP3 all had growth inhibition effects, the inhibition rate of vFV-HN-VP3 group was 51.7%, which was higher than that of vFV-HN, vFV-VP3 group and control group (p genes into mouse osteosarcoma cancer cells can cause cell a specificity anti-tumor immune activity, suppress tumor growth, and increase the survival rate of the tumor within host.

Protection against Foot-and-Mouth Disease Virus in Guinea Pigs via Oral Administration of Recombinant Lactobacillus plantarum Expressing VP1.

Directory of Open Access Journals (Sweden)

Miao Wang

Full Text Available Mucosal vaccination is an effective strategy for generating antigen-specific immune responses against mucosal infections of foot-and-mouth disease virus (FMDV. In this study, Lactobacillus plantarum strains NC8 and WCFS1 were used as oral delivery vehicles containing a pSIP411-VP1 recombinant plasmid to initiate mucosal and systemic immune responses in guinea pigs. Guinea pigs were orally vaccinated (three doses with NC8-pSIP411, NC8-pSIP411-VP1, WCFS1-pSIP411, WCFS1-pSIP411-VP1 or milk. Animals immunized with NC8-pSIP411-VP1 and WCFS1-pSIP411-VP1 developed high levels of antigen-specific serum IgG, IgA, IgM, mucosal secretory IgA (sIgA and neutralizing antibodies, and revealed stronger cell-mediated immune responses and enhanced protection against FMDV challenge compared with control groups. The recombinant pSIP411-VP1 effectively improved immunoprotection against FMDV in guinea pigs.

Video coding standards AVS China, H.264/MPEG-4 PART 10, HEVC, VP6, DIRAC and VC-1

CERN Document Server

Rao, K R; Hwang, Jae Jeong

2014-01-01

Review by Ashraf A. Kassim, Professor, Department of Electrical & Computer Engineering, and Associate Dean, School of Engineering, National University of Singapore.     The book consists of eight chapters of which the first two provide an overview of various video & image coding standards, and video formats. The next four chapters present in detail the Audio & video standard (AVS) of China, the H.264/MPEG-4 Advanced video coding (AVC) standard, High efficiency video coding (HEVC) standard and the VP6 video coding standard (now VP10) respectively. The performance of the wavelet based Dirac video codec is compared with H.264/MPEG-4 AVC in chapter 7. Finally in chapter 8, the VC-1 video coding standard is presented together with VC-2 which is based on the intra frame coding of Dirac and an outline of a H.264/AVC to VC-1 transcoder.   The authors also present and discuss relevant research literature such as those which document improved methods & techniques, and also point to other related reso...

Marburg virus VP35 can both fully coat the backbone and cap the ends of dsRNA for interferon antagonism.

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Shridhar Bale

2012-09-01

Full Text Available Filoviruses, including Marburg virus (MARV and Ebola virus (EBOV, cause fatal hemorrhagic fever in humans and non-human primates. All filoviruses encode a unique multi-functional protein termed VP35. The C-terminal double-stranded (dsRNA-binding domain (RBD of VP35 has been implicated in interferon antagonism and immune evasion. Crystal structures of the VP35 RBD from two ebolaviruses have previously demonstrated that the viral protein caps the ends of dsRNA. However, it is not yet understood how the expanses of dsRNA backbone, between the ends, are masked from immune surveillance during filovirus infection. Here, we report the crystal structure of MARV VP35 RBD bound to dsRNA. In the crystal structure, molecules of dsRNA stack end-to-end to form a pseudo-continuous oligonucleotide. This oligonucleotide is continuously and completely coated along its sugar-phosphate backbone by the MARV VP35 RBD. Analysis of dsRNA binding by dot-blot and isothermal titration calorimetry reveals that multiple copies of MARV VP35 RBD can indeed bind the dsRNA sugar-phosphate backbone in a cooperative manner in solution. Further, MARV VP35 RBD can also cap the ends of the dsRNA in solution, although this arrangement was not captured in crystals. Together, these studies suggest that MARV VP35 can both coat the backbone and cap the ends, and that for MARV, coating of the dsRNA backbone may be an essential mechanism by which dsRNA is masked from backbone-sensing immune surveillance molecules.

Directed chromosomal integration and expression of porcine rotavirus outer capsid protein VP4 in Lactobacillus casei ATCC393.

Science.gov (United States)

Yin, Ji-Yuan; Guo, Chao-Qun; Wang, Zi; Yu, Mei-Ling; Gao, Shuai; Bukhari, Syed M; Tang, Li-Jie; Xu, Yi-Gang; Li, Yi-Jing

2016-11-01

Using two-step plasmid integration in the presence of 5-fluorouracil (5-FU), we developed a stable and markerless Lactobacillus casei strain for vaccine antigen expression. The upp of L. casei, which encodes uracil phosphoribosyltransferase (UPRTase), was used as a counterselection marker. We employed the Δupp isogenic mutant, which is resistant to 5-FU, as host and a temperature-sensitive suicide plasmid bearing upp expression cassette as counterselectable integration vector. Extrachromosomal expression of UPRTase complemented the mutated chromosomal upp allele and restored sensitivity to 5-FU. The resultant genotype can either be wild type or recombinant. The efficacy of the system was demonstrated by insertion and expression of porcine rotavirus (PRV) VP4. To improve VP4 expression, we analyzed L. casei transcriptional profiles and selected the constitutive highly expressed enolase gene (eno). The VP4 inserted after the eno termination codon were screened in the presence of 5-FU. Using genomic PCR amplification, we confirmed that VP4 was successfully integrated and stably inherited for at least 50 generations. Western blot demonstrated that VP4 was steadily expressed in medium with different carbohydrates. RT-qPCR and ELISA analysis showed that VP4 expression from the chromosomal location was similar to that achieved by a plasmid expression system. Applying the recombinant strain to immunize BALB/c mice via oral administration revealed that the VP4-expressing L. casei could induce both specific local and systemic humoral immune responses in mice. Overall, the improved gene replacement system represents an efficient method for chromosome recombination in L. casei and provides a safe tool for vaccine production.

DNA repair rate and etoposide (VP16) resistance of tumor cell subpopulations derived from a single human small cell lung cancer

DEFF Research Database (Denmark)

Hansen, Lasse Tengbjerg; Lundin, Cecilia; Helleday, Thomas

2003-01-01

being VP16 resistant. In order to explain the VP16 resistant phenotype several mechanisms where considered. The p53 status, P-glycoprotein, MRP, topoisomerase IIalpha, and Mre11 protein levels, as well as growth kinetics, provided no explanations of the observed VP16 resistance. In contrast...

Controlling pore morphology and properties of nanoporous silica films using the different architecture PS-b-P2VP as a template.

Science.gov (United States)

Yu, Yang-Yen; Chien, Wen-Chen; Chen, Shih-Ting

2010-07-01

Nanoporous silica films were prepared through the templating of amphiphilic block copolymer, poly(styrene-2-vinyl pyridine) (PS-b-P2VP), and monodispersed colloidal silica nanoparticles. The experimental and theoretical studies suggested that the intermolecular hydrogen bonding existes between the colloidal silica nanoparticles and PS-b-P2VP. The effects of the loading ratio and P2VP chain length on the morphology and properties of the prepared nanoporous silica films were investigated. TEM and AFM studies showed that the uniform pore size could be achieved and the pore size increased with increasing porogen loading. The refractive index and dielectric constant of the prepared nanoporous films decreased with an increase in PS-b-P2VP loading. On the other hand, the porosity increased with an increasing PS-b-P2VP loading. This study demonstrated a methodology to control pore morphology and properties of the nanoporous silica films through the templating of PS-b-P2VP.

Radiation dose reduction using 100-kVp and a sinogram-affirmed iterative reconstruction algorithm in adolescent head CT: Impact on grey-white matter contrast and image noise

Energy Technology Data Exchange (ETDEWEB)

Nagayama, Yasunori [Kumamoto City Hospital, Department of Radiology, Kumamoto (Japan); Kumamoto University, Department of Diagnostic Radiology, Graduate School of Medical Sciences, Kumamoto (Japan); Nakaura, Takeshi; Yuki, Hideaki; Hirarta, Kenichiro; Kidoh, Masafumi; Oda, Seitaro; Utsunomiya, Daisuke; Yamashita, Yasuyuki [Kumamoto University, Department of Diagnostic Radiology, Graduate School of Medical Sciences, Kumamoto (Japan); Tsuji, Akinori; Urata, Joji; Furusawa, Mitsuhiro [Kumamoto City Hospital, Department of Radiology, Kumamoto (Japan)

2017-07-15

To retrospectively evaluate the image quality and radiation dose of 100-kVp scans with sinogram-affirmed iterative reconstruction (IR) for unenhanced head CT in adolescents. Sixty-nine patients aged 12-17 years underwent head CT under 120- (n = 34) or 100-kVp (n = 35) protocols. The 120-kVp images were reconstructed with filtered back-projection (FBP), 100-kVp images with FBP (100-kVp-F) and sinogram-affirmed IR (100-kVp-S). We compared the effective dose (ED), grey-white matter (GM-WM) contrast, image noise, and contrast-to-noise ratio (CNR) between protocols in supratentorial (ST) and posterior fossa (PS). We also assessed GM-WM contrast, image noise, sharpness, artifacts, and overall image quality on a four-point scale. ED was 46% lower with 100- than 120-kVp (p < 0.001). GM-WM contrast was higher, and image noise was lower, on 100-kVp-S than 120-kVp at ST (p < 0.001). CNR of 100-kVp-S was higher than of 120-kVp (p < 0.001). GM-WM contrast of 100-kVp-S was subjectively rated as better than of 120-kVp (p < 0.001). There were no significant differences in the other criteria between 100-kVp-S and 120-kVp (p = 0.072-0.966). The 100-kVp with sinogram-affirmed IR facilitated dramatic radiation reduction and better GM-WM contrast without increasing image noise in adolescent head CT. (orig.)

Crustal thickness and Vp/Vs beneath the southeastern United States: Constraints from receiver function stacking

Science.gov (United States)

Yang, Q.; Gao, S. S.; Liu, K. H.

2017-12-01

To provide new constraints on crustal structure and evolution models beneath a collage of tectonic provinces in the southeastern United States, a total of 10,753 teleseismic receiver functions recorded by 125 USArray and other seismic stations are used to compute crustal thickness and Vp/Vs values. The resulting crustal thicknesses range from 25 km at the coast to 51 km beneath the peak of the southern Appalachians with an average of 36.2 km ± 5.5 km. The resulting crustal thicknesses correlate well with surface elevation and Bouguer gravity anomalies. Beneath the Atlantic Coastal Plain, the crustal thicknesses show a clear eastward thinning with a magnitude of 10 km, from about 40 km beneath the western margin to 30 km beneath the coast. The Vp/Vs values for the entire study area range from 1.71 to 1.90 with a mean value of 1.80 ± 0.04. The mean Vp/Vs value is 1.82±0.035 in the southern Appalachian Mountain. The slightly larger than normal crustal Vp/Vs for this area might be the result of significant erosion of the felsic upper crust over the past 300 million years. Alternatively, it could also suggest the existence of pervasive magmatic intrusion into the Appalachian crust. The Vp/Vs measurements in the Atlantic Coastal Plain increase toward the east, ranging from 1.75 to 1.82, probably indicating a gradual increase of mafic magmatic intrusion into thinner crust during the development of the passive continental margin.

Expression of enterovirus 71 capsid protein VP1 in Escherichia coli and its clinical application

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Mei Shi

2013-12-01

Full Text Available The VPl gene of enterovirus 71 (EV71 was synthesized, construct a recombinant plasmid pET15b/VP1 and expressed in E. coli BL21. The recombinant VP1 protein could specifically react with EV71-infected patient sera without the cross-reaction with serum antibodies of coxsackievirus A16 (CA16, A4, A5, B3 and B5 as well as echovirus 6. In acute and convalescent phases, IgM and IgG antibodies of 182 serum samples were detected by ELISA with recombinant VP1 protein as a coated antigen. The results showed that the sensitivity, specificity, positive predictive value (PPV and negative predictive value (NPV of IgM antibodies in serum samples for the diagnosis of EV71 infection were 90.1, 98.4, 98.8 and 88.7%, respectively; similarly, those of IgG antibodies in serum samples were 82.4, 89.1, 91.5 and 78.1%, respectively. Five of 80 samples (6.25% from CA16infected patients were detected positive by ELISA with recombinant VP1 protein in which indicated the cross reactions and 0 of 5 samples from patients infected with other enteroviruses including CA4, CA5, CB3, CB5 and echovirus 6. Therefore, the recombinant VP1 protein of EV7l may provide a theoretical reference for establishing an effective antibody screening of IgM for EV71-infected patients with clinically suspected hand, foot, and mouth disease (HFMD.

[Prokaryotic expression of vp3 gene of Muscovy duck parvovirus, and its antiserum preparation for detection of virus multiplication].

Science.gov (United States)

Huang, Yu; Zhu, Yumin; Dong, Shijuan; Yu, Ruisong; Zhang, Yuanshu; Li, Zhen

2015-01-01

New epidemic broke out in recent year which was suspected to be caused by variant Muscovy duck parvovirus (MDPV). For this reason, new MDPV detection methods are needed for the new virus strains. In this study, a pair of primers were designed according to the full-length genome of MDPV strain SAAS-SHNH, which were identified in 2012, and were used to amplify the vp3 gene of MDPV by polymerase chain reaction. After being sequenced, the vp3 gene was subcloned into the prokaryotic expression vector PET28a. The recombinant plasmid was transformed into E. coli BL21 and induced with IPTG. SDS-PAGE and Western blotting analysis showed the MDPV vp3 gene was successfully expressed. After being purified by Ni2+ affinity chromatography system, the recombinant protein was used as antigen to immunize rabbits to obtain antiserum. Western blotting analysis showed that the acquired antiserum could react specifically with VP3 protein of J3D6 strain and MDPV vaccine strain. The antiserum could also be used for detection of cultured MDPV from primary duck embryo fibroblasts by immune fluorescence assay (IFA). It could be concluded that the VP3 protein and its antibody prepared in the research could be used for detection of VP3 antiserum and antigen respectively.

Neonatal treatment philosophy in Dutch and German NICUs: health-related quality of life in adulthood of VP/VLBW infants.

Science.gov (United States)

Breeman, Linda D; van der Pal, Sylvia; Verrips, Gijsbert H W; Baumann, Nicole; Bartmann, Peter; Wolke, Dieter

2017-04-01

Although survival after very preterm birth (VP)/very low birth weight (VLBW) has improved, a significant number of VP/VLBW individuals develop physical and cognitive problems during their life course that may affect their health-related quality of life (HRQoL). We compared HRQoL in VP/VLBW cohorts from two countries: The Netherlands (n = 314) versus Germany (n = 260) and examined whether different neonatal treatment and rates of disability affect HRQoL in adulthood. To analyse whether cohorts differed in adult HRQoL, linear regression analyses were performed for three HRQoL outcomes assessed with the Health Utilities Index 3 (HUI3), the London Handicap Scale (LHS), and the WHO Quality of Life instrument (WHOQOL-BREF). Stepwise hierarchical linear regression was used to test whether neonatal physical health and treatment, social environment, and intelligence (IQ) were related to VP/VLBW adults' HRQoL and cohort differences. Dutch VP/VLBW adults reported a significantly higher HRQoL on all three general HRQoL measures than German VP/VLBW adults (HUI3: .86 vs .83, p = .036; LHS: .93 vs. .90, p = .018; WHOQOL-BREF: 82.8 vs. 78.3, p life.

Immunogenicity of recombinant Lactobacillus plantarum NC8 expressing goose parvovirus VP2 gene in BALB/c mice.

Science.gov (United States)

Liu, Yu-Ying; Yang, Wen-Tao; Shi, Shao-Hua; Li, Ya-Jie; Zhao, Liang; Shi, Chun-Wei; Zhou, Fang-Yu; Jiang, Yan-Long; Hu, Jing-Tao; Gu, Wei; Yang, Gui-Lian; Wang, Chun-Feng

2017-06-30

Goose parvovirus (GPV) continues to be a threat to goose farms and has significant economic effects on the production of geese. Current commercially available vaccines only rarely prevent GPV infection. In our study, Lactobacillus (L.) plantarum NC8 was selected as a vector to express the VP2 gene of GPV, and recombinant L. plantarum pSIP409-VP2/NC8 was successfully constructed. The molecular weight of the expressed recombinant protein was approximately 70 kDa. Mice were immunized with a 2 × 10 9 colony-forming unit/200 μL dose of the recombinant L. plantarum strain, and the ratios and numbers of CD11c + , CD3 + CD4 + , CD3 + CD8 + , and interferon gamma- and tumor necrosis factor alpha-expressing spleen lymphocytes in the pSIP409-VP2/NC8 group were higher than those in the control groups. In addition, we assessed the capacity of L. plantarum SIP409-VP2/NC8 to induce secretory IgA production. We conclude that administered pSIP409-VP2/NC8 leads to relatively extensive cellular responses. This study provides information on GPV infection and offers a clear framework of options available for GPV control strategies.

GPM GROUND VALIDATION SATELLITE SIMULATED ORBITS C3VP V1

Data.gov (United States)

National Aeronautics and Space Administration — The GPM Ground Validation Satellite Simulated Orbits C3VP dataset is available in the Orbital database, which takes account for the atmospheric profiles, the...

Localization of VP28 on the baculovirus envelope and its immunogenicity against white spot syndrome virus in Penaeus monodon

International Nuclear Information System (INIS)

Syed Musthaq, S.; Madhan, Selvaraj; Sahul Hameed, A.S.; Kwang, Jimmy

2009-01-01

White spot syndrome virus (WSSV) is a large dsDNA virus responsible for white spot disease in shrimp and other crustaceans. VP28 is one of the major envelope proteins of WSSV and plays a crucial role in viral infection. In an effort to develop a vaccine against WSSV, we have constructed a recombinant baculovirus with an immediate early promoter 1 which expresses VP28 at an early stage of infection in insect cells. Baculovirus expressed rVP28 was able to maintain its structural and antigenic conformity as indicated by immunofluorescence assay and western blot analysis. Interestingly, our results with confocal microscopy revealed that rVP28 was able to localize on the plasma membrane of insect cells infected with recombinant baculovirus. In addition, we demonstrated with transmission electron microscopy that baculovirus successfully acquired rVP28 from the insect cell membrane via the budding process. Using this baculovirus displaying VP28 as a vaccine against WSSV, we observed a significantly higher survival rate of 86.3% and 73.5% of WSSV-infected shrimp at 3 and 15 days post vaccination respectively. Quantitative real-time PCR also indicated that the WSSV viral load in vaccinated shrimp was significantly reduced at 7 days post challenge. Furthermore, our RT-PCR and immunohistochemistry results demonstrated that the recombinant baculovirus was able to express VP28 in vivo in shrimp tissues. This study will be of considerable significance in elucidating the morphogenesis of WSSV and will pave the way for new generation vaccines against WSSV.

Role of a nuclear localization signal on the minor capsid Proteins VP2 and VP3 in BKPyV nuclear entry

Energy Technology Data Exchange (ETDEWEB)

Bennett, Shauna M. [Cellular and Molecular Biology Program University of Michigan 1150W Medical Center Dr 5724 Medical Science Bldg II Ann Arbor, MI 48109 (United States); Zhao, Linbo [Doctoral Program in Cancer Biology Program University of Michigan 1150W Medical Center Dr 5724 Medical Science Bldg II Ann Arbor, MI 48109 (United States); Bosard, Catherine [Department of Microbiology and Immunology University of Michigan 1150W Medical Center Dr 5724 Medical Science Bldg II Ann Arbor, MI 48109 (United States); Imperiale, Michael J., E-mail: imperial@umich.edu [Cellular and Molecular Biology Program University of Michigan 1150W Medical Center Dr 5724 Medical Science Bldg II Ann Arbor, MI 48109 (United States); Doctoral Program in Cancer Biology Program University of Michigan 1150W Medical Center Dr 5724 Medical Science Bldg II Ann Arbor, MI 48109 (United States); Department of Microbiology and Immunology University of Michigan 1150W Medical Center Dr 5724 Medical Science Bldg II Ann Arbor, MI 48109 (United States)

2015-01-01

BK Polyomavirus (BKPyV) is a ubiquitous nonenveloped human virus that can cause severe disease in immunocompromised populations. After internalization into renal proximal tubule epithelial cells, BKPyV traffics through the ER and enters the cytosol. However, it is unclear how the virus enters the nucleus. In this study, we elucidate a role for the nuclear localization signal located on the minor capsid proteins VP2 and VP3 during infection. Site-directed mutagenesis of a single lysine in the basic region of the C-terminus of the minor capsid proteins abrogated their nuclear localization, and the analogous genomic mutation reduced infectivity. Additionally, through use of the inhibitor ivermectin and knockdown of importin β1, we found that the importin α/β pathway is involved during infection. Overall these data are the first to show the significance of the NLS of the BKPyV minor capsid proteins during infection in a natural host cell. - Highlights: • Polyomaviruses must deliver their genome to the nucleus to replicate. • The minor capsid proteins have a well-conserved nuclear localization signal. • Mutation of this NLS diminishes, but does not completely inhibit, infection.

Role of a nuclear localization signal on the minor capsid Proteins VP2 and VP3 in BKPyV nuclear entry

International Nuclear Information System (INIS)

Bennett, Shauna M.; Zhao, Linbo; Bosard, Catherine; Imperiale, Michael J.

2015-01-01

BK Polyomavirus (BKPyV) is a ubiquitous nonenveloped human virus that can cause severe disease in immunocompromised populations. After internalization into renal proximal tubule epithelial cells, BKPyV traffics through the ER and enters the cytosol. However, it is unclear how the virus enters the nucleus. In this study, we elucidate a role for the nuclear localization signal located on the minor capsid proteins VP2 and VP3 during infection. Site-directed mutagenesis of a single lysine in the basic region of the C-terminus of the minor capsid proteins abrogated their nuclear localization, and the analogous genomic mutation reduced infectivity. Additionally, through use of the inhibitor ivermectin and knockdown of importin β1, we found that the importin α/β pathway is involved during infection. Overall these data are the first to show the significance of the NLS of the BKPyV minor capsid proteins during infection in a natural host cell. - Highlights: • Polyomaviruses must deliver their genome to the nucleus to replicate. • The minor capsid proteins have a well-conserved nuclear localization signal. • Mutation of this NLS diminishes, but does not completely inhibit, infection

An argument for VP coordination: scene-setting coordination and ...

African Journals Online (AJOL)

This article demonstrates the properties of this curious construction type and proposes the first analysis to date. It is argued that this is an instance of VP coordination and that this configuration allows the possibility of high merger of direct objects in a constrained fashion. Southern African Linguistics and Applied Language ...

Evaluation of Breast Cancer Awareness among Women Presenting ...

African Journals Online (AJOL)

Conclusion: This study has revealed low breast cancer awareness among women presenting with newly diagnosed breast disease at the Universitas Hospital, and hence low rates of self- and clinical examinations of the breast and low mammographic screening rates. This may be attributed to a general lack of awareness of ...

Identification of H-2d Restricted T Cell Epitope of Foot-and-mouth Disease Virus Structural Protein VP1

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Zhang Zhong-Wang

2011-09-01

Full Text Available Abstract Background Foot-and-mouth disease (FMD is a highly contagious and devastating disease affecting livestock that causes significant financial losses. Therefore, safer and more effective vaccines are required against Foot-and-mouth disease virus(FMDV. The purpose of this study is to screen and identify an H-2d restricted T cell epitope from the virus structural protein VP1, which is present with FMD. We therefore provide a method and basis for studying a specific FMDV T cell epitope. Results A codon-optimized expression method was adopted for effective expression of VP1 protein in colon bacillus. We used foot-and-mouth disease standard positive serum was used for Western blot detection of its immunogenicity. The VP1 protein was used for immunizing BALB/c mice, and spleen lymphocytes were isolated. Then, a common in vitro training stimulus was conducted for potential H-2Dd, H-2Kd and H-2Ld restricted T cell epitope on VP1 proteins that were predicted and synthesized by using a bioinformatics method. The H-2Kd restricted T cell epitope pK1 (AYHKGPFTRL and the H-2Dd restricted T cell epitope pD7 (GFIMDRFVKI were identified using lymphocyte proliferation assays and IFN-γ ELISPOT experiments. Conclusions The results of this study lay foundation for studying the FMDV immune process, vaccine development, among other things. These results also showed that, to identify viral T cell epitopes, the combined application of bioinformatics and molecular biology methods is effective.

Clinical presentation of urinary tract infection (UTI) differs with aging in women.

Science.gov (United States)

Arinzon, Zeev; Shabat, Shay; Peisakh, Alexander; Berner, Yitshal

2012-01-01

Uncomplicated UTI is among the most common health problems seen in general practice and typically affects immunocompetent, anatomically normal women. The aim of this study was to explore the difference in clinical presentation in acute, uncomplicated UTI in otherwise healthy community dwelling, premenopausal (Pre-M) and postmenopausal (Post-M) women. A UTI was defined as uropathogen of more than 10(3)cfu/ml in midstream urine culture. Symptoms of UTI were divided to three: during voiding, local symptoms, and generalized symptoms. A total of 196 women aged a minimum of 45 years with diagnosis of UTI were studied. The patients were divided into two groups: Pre-M (n=102, mean age 48.14 years) and Post-M (n=94, mean age 69.21 years). The predominant complaints in Pre-M women were local symptoms. The clinical presentations showed more severity in the Post-M group than in Pre-M women, predominantly generalized unspecific symptoms and storage symptoms. Advanced age positively correlated with urgency of urination, painful voiding, urinary incontinence, sexual activity, low-back pain, lower abdominal pain and negatively correlated with frequency, painful and burning of urination and bladder pain. Our study showed that clinical presentation of UTI in Pre-M and Post-M women is different. The differences are presented not only by the voiding itself and by local symptoms but also by unspecified generalized symptoms that is especially important in elderly patients. Copyright © 2011. Published by Elsevier Ireland Ltd.

Emotional distress in women presenting for breast imaging

International Nuclear Information System (INIS)

Gupta, R.; Roy, S.; Nayak, Madhabika B.; Khoursheed, M.

1999-01-01

The aim of this study was to assess anxiety and depression in a sample of women presenting for imaging of breast following a clinical referral. Emotional distress in the women was also assessed in relation to demographic factors, reason for referral, presence for breast symptoms, type of imaging procedure performed and self-reported pain and discomfort during imaging. The study comprised 167 patients. The Hopkins Symptom Checklist-25 (HSCL-25) and a discomfort rating scale were used to assess emotional distress and discomfort or pain experienced during the imaging. While less than 10% of all subjects scored above psychiatric cut-off points for anxiety and depression, 25% and 20% reported significant distress associated with anxiety and depression symptoms respectively. Education alone was associated with higher anxiety scores, while the presence of breast symptoms significantly increased depression scores and reports of specific nonsomatic symptoms of depression. Higher anxiety and depression scores were also associated with pain experienced during the imaging procedure. Emotional distress may negatively impact women's experience of breast imaging. Screening for emotional distress is important within the context of breast imaging. (author)

Identification of novel Ebola virus (EBOV) VP24 inhibitor from Indonesian natural products through in silico drug design approach

Science.gov (United States)

Tambunan, U. S. F.; Nasution, M. A. F.

2017-07-01

Ebola remains as one of the deadliest diseases in the world, with almost 29,000 cases were reported and kill 11,000 of them, and yet neither treatment nor vaccine that can combat this disease effectively. This disease is caused by ebolavirus (EBOV), a primary member of Filoviridae family. The life cycle of this virus has been operated by several key proteins, one of them is VP24 protein, which has been known for its crucial role in the transcription and replication of EBOV. Therefore, targeting VP24 protein can be a solution for treating this pathogenic disease. In this study, virtual screening of Indonesian natural products as EBOV VP24 inhibitor was performed. About 2,020 ligands from many sources, including HerbalDB database, were obtained and screened by using DataWarrior software to measure its molecular and pharmacological properties, resulting 301 ligands in the process. Then, the molecular docking simulation was performed to check the ligand's binding interaction and affinity with EBOV VP24 protein; this simulation was done by using MOE 2014.09 software. This study resulted that cycloartocarpin was the best ligand to inhibit the EBOV VP24 protein. Therefore, this ligand should be checked its stability through molecular dynamics simulation and performed in vitro test to verify its bioactivity against the EBOV VP24 protein.

The Derivation of Scandinavian Object Shift and Remnant VP-Topicalisation

DEFF Research Database (Denmark)

Engels, Eva; Vikner, Sten

2013-01-01

Based on the examination of remnant VP-topicalization constructions, this chapter argues for an order preservation analysis to Scandinavian Object Shift. Reviewing Fox and Pesetsky’s (2003, 2005) cyclic linearization approach and extending the empirical data base, we show that the phenomena are b...

Differences in Presentation, Management and Outcomes in Women and Men Presenting to an Emergency Department With Possible Cardiac Chest Pain.

Science.gov (United States)

Ruane, Lorcan; H Greenslade, Jaimi; Parsonage, William; Hawkins, Tracey; Hammett, Christopher; Lam, Carolyn Sp; Knowlman, Thomas; Doig, Shaela; Cullen, Louise

2017-12-01

Research suggests that female patients with acute coronary syndrome (ACS) experience delays in emergency department (ED) management and are less likely to receive guideline-based treatments and referrals for follow-up testing. Women are often found to have poorer clinical outcomes in comparison to men. This study aimed to assess current sex differences in the presentation, management and outcomes of patients with undifferentiated chest pain presenting to a tertiary ED. Data were analysed from two prospective studies conducted at a single Australian site between 2007 and 2014. Eligible patients were those of 18 years of age or older presenting with at least 5 minutes of chest pain or other symptoms for which the treating physician planned to investigate for possible ACS. Presenting symptoms, ED time measures, follow-up testing and outcomes, including 30-day ACS and mortality, were measured and compared between male and female patients. Of 2349 (60% men) patients presenting with chest pain, 153 men and 51 women were diagnosed with ACS within 30days . Presenting symptoms were similar in men and women with confirmed ACS. Time from symptom onset to ED presentation, time spent in the ED and total time in hospital were similar between the sexes. Male and female patients had similar rates of follow-up provocative testing. After adjustment for clinical factors, the odds of undergoing angiography were 1.8 (95% CI: 1.36-2.40) times higher for men than women. Of those undergoing coronary angiography within 30 days, a smaller proportion of women, compared to men, received revascularisation. Within 30 days, three (0.2%) male and one (0.1%) female patient died. Minimal sex differences were observed in the contemporary emergency management of patients presenting with suspected ACS. Thirty-day outcomes were similarly low in men and women despite lower rates of coronary angiography and revascularisation in women. Further research is required to replicate these results in different

Evaluation of breast cancer awareness among women presenting ...

African Journals Online (AJOL)

leading cause of death from cancer among South African females.1,2. Of significance at present is the ... that the few young black women who get breast cancer are more likely ... June 2006. The results of the three-month breast cancer awareness campaign .... It does, however, reflect negatively on the general breast cancer ...

Quality of image and exposure dose according to kVp, mA and lterative reconstruction in computed tomography

Energy Technology Data Exchange (ETDEWEB)

Cha, Sang Young [Dept. of Radiology, Inha University Hospital, Incheon (Korea, Republic of); Park, Jae Yoon [Dept. of Radiology, Incheon Christian Hospital, Incheon (Korea, Republic of); Lee, Yong Ki [Dept. of Radiological Technology, DongNam Health University, Suwon (Korea, Republic of); Kim, Jeon Hun [Dept. of Radiatioin Oncology, Konyang University Hospital, Daejeon (Korea, Republic of); Choi, Jae Ho [Dept. of Radiological Technology, Ansan University, Ansan (Korea, Republic of)

2017-09-15

The purpose of this study is to investigate the image quality and exposure dose according to kVp and mAs in CT and to confirm improvement in image quality according to None IR and IR(Iterative Reconstruction) levels. Measurement results of image quality using Image J, HU(Hounsfield units) and BN(Background Noise) are decreased, while SNR(Signal to Noise Ratio) and CTDIvol(CT dose index volume) are increased as the kVp increases and there was no change of BHU(Background Hounsfield units). BN was reduced due to increased kVp, while SNR and CTDIvol were increased. Also, the higher IR stage, the lower BN, SI(Signal Intensity) and HU while SNR was improved by about 10∼60%. Based on this, when applying IR for clinical applications, it is necessary to finely adjust kVp and mA with a phased approach.

Putative endogenous filovirus VP35-like protein potentially functions as an IFN antagonist but not a polymerase cofactor.

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Tatsunari Kondoh

Full Text Available It has been proposed that some non-retroviral RNA virus genes are integrated into vertebrate genomes. Endogenous filovirus-like elements (EFLs have been discovered in some mammalian genomes. However, their potential roles in ebolavirus infection are unclear. A filovirus VP35-like element (mlEFL35 is found in the little brown bat (Myotis lucifugus genome. Putative mlEFL35-derived protein (mlEFL35p contains nearly full-length amino acid sequences corresponding to ebolavirus VP35. Ebola virus VP35 has been shown to bind double-stranded RNA, leading to inhibition of type I interferon (IFN production, and is also known as a viral polymerase cofactor that is essential for viral RNA transcription/replication. In this study, we transiently expressed mlEFL35p in human kidney cells and investigated its biological functions. We first found that mlEFL35p was coimmunoprecipitated with itself and ebolavirus VP35s but not with the viral nucleoprotein. Then the biological functions of mlEFL35p were analyzed by comparing it to ebolavirus VP35s. We found that the expression of mlEFL35p significantly inhibited human IFN-β promoter activity as well as VP35s. By contrast, expression of mlEFL35p did not support viral RNA transcription/replication and indeed slightly decrease the reporter gene expression in a minigenome assay. These results suggest that mlEFL35p potentially acts as an IFN antagonist but not a polymerase cofactor.

Image quality, radiation dose and diagnostic accuracy of 70 kVp whole brain volumetric CT perfusion imaging: a preliminary study

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Fang, Xiao Kun; Ni, Qian Qian; Zhou, Chang Sheng; Chen, Guo Zhong; Luo, Song; Zhang, Long Jiang; Lu, Guang Ming [Medical School of Nanjing University, Department of Medical Imaging, Jinling Hospital, Nanjing, Jiangsu (China); Schoepf, U.J. [Medical School of Nanjing University, Department of Medical Imaging, Jinling Hospital, Nanjing, Jiangsu (China); Medical University of South Carolina, Ashley River Tower, Division of Cardiovascular Imaging, Charleston, SC (United States); Fuller, Stephen R.; De Cecco, Carlo N. [Medical University of South Carolina, Ashley River Tower, Division of Cardiovascular Imaging, Charleston, SC (United States)

2016-11-15

To evaluate image quality and diagnostic accuracy for acute infarct detection and radiation dose of 70 kVp whole brain CT perfusion (CTP) and CT angiography (CTA) reconstructed from CTP source data. Patients were divided into three groups (n = 50 each): group A, 80 kVp, 21 scanning time points; groups B, 70 kVp, 21 scanning time points; group C, 70 kVp, 17 scanning time points. Objective and subjective image quality of CTP and CTA were compared. Diagnostic accuracy for detecting acute infarct and cerebral artery stenosis ≥ 50 % was calculated for CTP and CTA with diffusion weighted imaging and digital subtraction angiography as reference standards. Effective radiation dose was compared. There were no differences in any perfusion parameter value between three groups (P > 0.05). No difference was found in subjective image quality between three groups (P > 0.05). Diagnostic accuracy for detecting acute infarct and vascular stenosis showed no difference between three groups (P > 0.05). Compared with group A, radiation doses of groups B and C were decreased by 28 % and 37 % (both P < 0.001), respectively. Compared with 80 kVp protocol, 70 kVp brain CTP allows comparable vascular and perfusion assessment and lower radiation dose while maintaining high diagnostic accuracy in detecting acute infarct. (orig.)

Identification of Rotavirus VP6-Specific CD4+ T Cell Epitopes in a G1P[8] Human Rotavirus-Infected Rhesus Macaque

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Wei Zhao

2008-01-01

Full Text Available A non-human primate model was used to evaluate its potential for identification of rotavirus viral protein 6 (VP6 CD4+ T cell epitopes. Four juvenile rhesus macaques were inoculated with a mixed inoculum (G1P[8] and G9P[8] of human rotaviruses. Infection accompanied by G1P[8] shedding was achieved in the two macaques that had no rotavirus immunoglobulin A (IgA in plasma. To measure the interferon gamma (IFN-γ and tumor necrosis factor (TNF anti-viral cytokines produced by peripheral CD4+ cells that recognize VP6 epitopes, whole blood cells from one infected macaque were stimulated in vitro with VP6 peptides. Stimulation with peptide pools derived from the simian rotavirus VP6 161–395 region revealed reactivity of CD4+ T cells with the VP6 281–331 domain. A VP6 301–315 region was identified as the epitope responsible for IFN-γ production while a broader VP6 293–327 domain was linked to TNF production. These results suggest that human rotavirus-infected macaques can be used for identification of additional epitopes and domains to address specific questions related to the development of pediatric vaccines.

Eclipse Phase of Herpes Simplex Virus Type 1 Infection: Efficient Dynein-Mediated Capsid Transport without the Small Capsid Protein VP26

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Döhner, Katinka; Radtke, Kerstin; Schmidt, Simone; Sodeik, Beate

2006-01-01

Cytoplasmic dynein,together with its cofactor dynactin, transports incoming herpes simplex virus type 1 (HSV-1) capsids along microtubules (MT) to the MT-organizing center (MTOC). From the MTOC, capsids move further to the nuclear pore, where the viral genome is released into the nucleoplasm. The small capsid protein VP26 can interact with the dynein light chains Tctex1 (DYNLT1) and rp3 (DYNLT3) and may recruit dynein to the capsid. Therefore, we analyzed nuclear targeting of incoming HSV1-ΔVP26 capsids devoid of VP26 and of HSV1-GFPVP26 capsids expressing a GFPVP26 fusion instead of VP26. To compare the cell entry of different strains, we characterized the inocula with respect to infectivity, viral genome content, protein composition, and particle composition. Preparations with a low particle-to-PFU ratio showed efficient nuclear targeting and were considered to be of higher quality than those containing many defective particles, which were unable to induce plaque formation. When cells were infected with HSV-1 wild type, HSV1-ΔVP26, or HSV1-GFPVP26, viral capsids were transported along MT to the nucleus. Moreover, when dynein function was inhibited by overexpression of the dynactin subunit dynamitin, fewer capsids of HSV-1 wild type, HSV1-ΔVP26, and HSV1-GFPVP26 arrived at the nucleus. Thus, even in the absence of the potential viral dynein receptor VP26, HSV-1 used MT and dynein for efficient nuclear targeting. These data suggest that besides VP26, HSV-1 encodes other receptors for dynein or dynactin. PMID:16873277

[V.P. Kravkov as a sanitary physician of the Russian Imperial Army (to centenary of beginning of the First World War)].

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Uzbekova, D G

2014-01-01

The article presents for the first time the description of life story and service of V.P. Kravkov (1859-1920)--participant of Russian-Japanese war and the First World War, doctor of medicine, sanitary physician of the Russian Imperial Army and author of number of books and articles on preventive medicine. During these two wars, V.P. Kravkov organized sanitary epidemiological service in fronts. He kept diary regularly reflecting conditions of sanitary epidemiological service during this period, describing one's own observations and assessing survived events. The manuscripts of diaries are stored in the Russian state library. The diaries made great input into summarizing of experience of the Russian military sanitary service and became valuable monument of military memoirs' literature.

Role of protein phosphatase 1 in dephosphorylation of Ebola virus VP30 protein and its targeting for the inhibition of viral transcription.

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Ilinykh, Philipp A; Tigabu, Bersabeh; Ivanov, Andrey; Ammosova, Tatiana; Obukhov, Yuri; Garron, Tania; Kumari, Namita; Kovalskyy, Dmytro; Platonov, Maxim O; Naumchik, Vasiliy S; Freiberg, Alexander N; Nekhai, Sergei; Bukreyev, Alexander

2014-08-15

The filovirus Ebola (EBOV) causes the most severe hemorrhagic fever known. The EBOV RNA-dependent polymerase complex includes a filovirus-specific VP30, which is critical for the transcriptional but not replication activity of EBOV polymerase; to support transcription, VP30 must be in a dephosphorylated form. Here we show that EBOV VP30 is phosphorylated not only at the N-terminal serine clusters identified previously but also at the threonine residues at positions 143 and 146. We also show that host cell protein phosphatase 1 (PP1) controls VP30 dephosphorylation because expression of a PP1-binding peptide cdNIPP1 increased VP30 phosphorylation. Moreover, targeting PP1 mRNA by shRNA resulted in the overexpression of SIPP1, a cytoplasm-shuttling regulatory subunit of PP1, and increased EBOV transcription, suggesting that cytoplasmic accumulation of PP1 induces EBOV transcription. Furthermore, we developed a small molecule compound, 1E7-03, that targeted a non-catalytic site of PP1 and increased VP30 dephosphorylation. The compound inhibited the transcription but increased replication of the viral genome and completely suppressed replication of EBOV in cultured cells. Finally, mutations of Thr(143) and Thr(146) of VP30 significantly inhibited EBOV transcription and strongly induced VP30 phosphorylation in the N-terminal Ser residues 29-46, suggesting a novel mechanism of regulation of VP30 phosphorylation. Our findings suggest that targeting PP1 with small molecules is a feasible approach to achieve dysregulation of the EBOV polymerase activity. This novel approach may be used for the development of antivirals against EBOV and other filovirus species. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

Oral immunization with rotavirus VP7 expressed in transgenic potatoes induced high titers of mucosal neutralizing IgA

International Nuclear Information System (INIS)

Wu Yuzhang; Li Jintao; Mou Zhirong; Fei Lei; Ni Bing; Geng Miao; Jia Zhengcai; Zhou Wei; Zou Liyun; Tang Yan

2003-01-01

Rotaviruses (RV) are a common cause of severe diarrhea in young children, resulting in nearly one million deaths worldwide annually. Rotavirus VP7 was the rotavirus neutralizing protein. Previous study reported that VP7 DNA vaccine can induce high levels of IgG in mice but cannot protect mice against challenge (Choi, A.H., Basu, M., Rae, M.N., McNeal, M.M., Ward, R.L., 1998. Virology 250, 230-240). We found that rotavirus VP7 could maintain its neutralizing immunity when it was transformed into the potato genome. Mice immunized with the transformed tubers successfully elicited serum IgG and mucosal IgA specific for VP7. The mucosal IgA titer was as high as 1000, while serum IgG titer was only 600. Neutralizing assays indicated that IgA could neutralize rotavirus. These results indicate the potential usefulness of plants for production and delivery of edible rotavirus vaccines

Multiplicity distributions of charged hadrons in vp and charged current interactions

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Jones, G. T.; Jones, R. W. L.; Kennedy, B. W.; Morrison, D. R. O.; Mobayyen, M. M.; Wainstein, S.; Aderholz, M.; Hantke, D.; Katz, U. F.; Kern, J.; Schmitz, N.; Wittek, W.; Borner, H. P.; Myatt, G.; Radojicic, D.; Burke, S.

1992-03-01

Using data on vp andbar vp charged current interactions from a bubble chamber experiment with BEBC at CERN, the multiplicity distributions of charged hadrons are investigated. The analysis is based on ˜20000 events with incident v and ˜10000 events with incidentbar v. The invariant mass W of the total hadronic system ranges from 3 GeV to ˜14 GeV. The experimental multiplicity distributions are fitted by the binomial function (for different intervals of W and in different intervals of the rapidity y), by the Levy function and the lognormal function. All three parametrizations give acceptable values for X 2. For fixed W, forward and backward multiplicities are found to be uncorrelated. The normalized moments of the charged multiplicity distributions are measured as a function of W. They show a violation of KNO scaling.

Development and Validation of a Novel Dual Luciferase Reporter Gene Assay to Quantify Ebola Virus VP24 Inhibition of IFN Signaling

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Elisa Fanunza

2018-02-01

Full Text Available The interferon (IFN system is the first line of defense against viral infections. Evasion of IFN signaling by Ebola viral protein 24 (VP24 is a critical event in the pathogenesis of the infection and, hence, VP24 is a potential target for drug development. Since no drugs target VP24, the identification of molecules able to inhibit VP24, restoring and possibly enhancing the IFN response, is a goal of concern. Accordingly, we developed a dual signal firefly and Renilla luciferase cell-based drug screening assay able to quantify IFN-mediated induction of Interferon Stimulated Genes (ISGs and its inhibition by VP24. Human Embryonic Kidney 293T (HEK293T cells were transiently transfected with a luciferase reporter gene construct driven by the promoter of ISGs, Interferon-Stimulated Response Element (ISRE. Stimulation of cells with IFN-α activated the IFN cascade leading to the expression of ISRE. Cotransfection of cells with a plasmid expressing VP24 cloned from a virus isolated during the last 2014 outbreak led to the inhibition of ISRE transcription, quantified by a luminescent signal. To adapt this system to test a large number of compounds, we performed it in 96-well plates; optimized the assay analyzing different parameters; and validated the system by calculating the Z′- and Z-factor, which showed values of 0.62 and 0.53 for IFN-α stimulation assay and VP24 inhibition assay, respectively, indicative of robust assay performance.

Renal Cyst Pseudoenhancement: Intraindividual Comparison Between Virtual Monochromatic Spectral Images and Conventional Polychromatic 120-kVp Images Obtained During the Same CT Examination and Comparisons Among Images Reconstructed Using Filtered Back Projection, Adaptive Statistical Iterative Reconstruction, and Model-Based Iterative Reconstruction

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Yamada, Yoshitake; Yamada, Minoru; Sugisawa, Koichi; Akita, Hirotaka; Shiomi, Eisuke; Abe, Takayuki; Okuda, Shigeo; Jinzaki, Masahiro

2015-01-01

Abstract The purpose of this study was to compare renal cyst pseudoenhancement between virtual monochromatic spectral (VMS) and conventional polychromatic 120-kVp images obtained during the same abdominal computed tomography (CT) examination and among images reconstructed using filtered back projection (FBP), adaptive statistical iterative reconstruction (ASIR), and model-based iterative reconstruction (MBIR). Our institutional review board approved this prospective study; each participant provided written informed consent. Thirty-one patients (19 men, 12 women; age range, 59–85 years; mean age, 73.2 ± 5.5 years) with renal cysts underwent unenhanced 120-kVp CT followed by sequential fast kVp-switching dual-energy (80/140 kVp) and 120-kVp abdominal enhanced CT in the nephrographic phase over a 10-cm scan length with a random acquisition order and 4.5-second intervals. Fifty-one renal cysts (maximal diameter, 18.0 ± 14.7 mm [range, 4–61 mm]) were identified. The CT attenuation values of the cysts as well as of the kidneys were measured on the unenhanced images, enhanced VMS images (at 70 keV) reconstructed using FBP and ASIR from dual-energy data, and enhanced 120-kVp images reconstructed using FBP, ASIR, and MBIR. The results were analyzed using the mixed-effects model and paired t test with Bonferroni correction. The attenuation increases (pseudoenhancement) of the renal cysts on the VMS images reconstructed using FBP/ASIR (least square mean, 5.0/6.0 Hounsfield units [HU]; 95% confidence interval, 2.6–7.4/3.6–8.4 HU) were significantly lower than those on the conventional 120-kVp images reconstructed using FBP/ASIR/MBIR (least square mean, 12.1/12.8/11.8 HU; 95% confidence interval, 9.8–14.5/10.4–15.1/9.4–14.2 HU) (all P < .001); on the other hand, the CT attenuation values of the kidneys on the VMS images were comparable to those on the 120-kVp images. Regardless of the reconstruction algorithm, 70-keV VMS images showed

Identification and production of mouse scFv to specific epitope of enterovirus-71 virion protein-2 (VP2).

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Thanongsaksrikul, Jeeraphong; Srimanote, Potjanee; Tongtawe, Pongsri; Glab-Ampai, Kittirat; Malik, Aijaz Ahmad; Supasorn, Oratai; Chiawwit, Phatcharaporn; Poovorawan, Yong; Chaicumpa, Wanpen

2018-05-01

Enterovirus-71 (EV71) and coxsackievirus-A16 (CA16) frequently cause hand-foot-mouth disease (HFMD) epidemics among infants and young children. CA16 infections are usually mild, while EV71 disease may be fatal due to neurologic complications. As such, the ability to rapidly and specifically recognize EV71 is needed to facilitate proper case management and epidemic control. Accordingly, the aim of this study was to generate antibodies to EV71-virion protein-2 (VP2) by phage display technology for further use in specific detection of EV71. A recombinant peptide sequence of EV71-VP2, carrying a predicted conserved B cell epitope fused to glutathione-S-transferase (GST) (designated GST-EV71-VP2/131-160), was produced. The fusion protein was used as bait in in-solution biopanning to separate protein-bound phages from a murine scFv (MuscFv) phage display library constructed from an immunoglobulin gene repertoire from naïve ICR mice. Three phage-transformed E. coli clones (clones 63, 82, and 83) produced MuscFvs that bound to the GST-EV71-VP2/131-160 peptide. The MuscFv of clone 83 (MuscFv83), which produced the highest ELISA signal to the target antigen, was further tested. MuscFv83 also bound to full-length EV71-VP2 and EV71 particles, but did not bind to GST, full-length EV71-VP1, or the antigenically related CA16. MuscFv83 could be a suitable reagent for rapid antigen-based immunoassay, such as immunochromatography (ICT), for the specific detection and/or diagnosis of EV71 infection as well as epidemic surveillance.

Hydatidiform mole: age-related clinical presentation and high rate of severe complications in older women.

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Mangili, Giorgia; Giorgione, Veronica; Gentile, Cinzia; Bergamini, Alice; Pella, Francesca; Almirante, Giada; Candiani, Massimo

2014-05-01

The purpose of this study was to demonstrate differences in clinical presentation of hydatidiform mole between women ≥40 years and younger women. Retrospective study. A tertiary referral unit in northern Italy. Three hundred and sixty-five women with hydatidiform mole were divided into group A (Clinical presentation between groups A and B was analyzed, also considering partial hydatidiform mole and complete hydatidiform mole. Differences in clinical presentation according to woman's age. In group B the diagnosis of hydatidiform mole at ≥12 gestational weeks was more frequent (p presented with vaginal bleeding (p clinical features of hydatidiform mole in women ≥40 years are different from those seen in younger women. Failures in the early detection of hydatidiform mole in older women may expose them to a higher rate of severe complications. © 2014 Nordic Federation of Societies of Obstetrics and Gynecology.

Crystallization and preliminary X-ray diffraction analysis of the sialic acid-binding domain (VP8*) of porcine rotavirus strain CRW-8

International Nuclear Information System (INIS)

Scott, Stacy A.; Holloway, Gavan; Coulson, Barbara S.; Szyczew, Alex J.; Kiefel, Milton J.; Itzstein, Mark von; Blanchard, Helen

2005-01-01

The sialic acid-binding domain (VP8*) component of the porcine CRW-8 rotavirus spike protein has been overexpressed in E. coli, purified and co-crystallized with an N-acetylneuraminic acid derivative. X-ray diffraction data have been collected to 2.3 Å, which has enabled determination of the structure by molecular replacement. Rotavirus recognition and attachment to host cells involves interaction with the spike protein VP4 that projects outwards from the surface of the virus particle. An integral component of these spikes is the VP8* domain, which is implicated in the direct recognition and binding of sialic acid-containing cell-surface carbohydrates and facilitates subsequent invasion by the virus. The expression, purification, crystallization and preliminary X-ray diffraction analysis of VP8* from porcine CRW-8 rotavirus is reported. Diffraction data have been collected to 2.3 Å resolution, enabling the determination of the VP8* structure by molecular replacement

Crystallization and preliminary X-ray diffraction analysis of the sialic acid-binding domain (VP8*) of porcine rotavirus strain CRW-8

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Scott, Stacy A. [Institute for Glycomics, Griffith University (Gold Coast Campus) PMB 50, Gold Coast Mail Centre, Queensland 9726 (Australia); Holloway, Gavan; Coulson, Barbara S. [Department of Microbiology and Immunology, The University of Melbourne, Victoria 3010 (Australia); Szyczew, Alex J.; Kiefel, Milton J.; Itzstein, Mark von; Blanchard, Helen, E-mail: h.blanchard@griffith.edu.au [Institute for Glycomics, Griffith University (Gold Coast Campus) PMB 50, Gold Coast Mail Centre, Queensland 9726 (Australia)

2005-06-01

The sialic acid-binding domain (VP8*) component of the porcine CRW-8 rotavirus spike protein has been overexpressed in E. coli, purified and co-crystallized with an N-acetylneuraminic acid derivative. X-ray diffraction data have been collected to 2.3 Å, which has enabled determination of the structure by molecular replacement. Rotavirus recognition and attachment to host cells involves interaction with the spike protein VP4 that projects outwards from the surface of the virus particle. An integral component of these spikes is the VP8* domain, which is implicated in the direct recognition and binding of sialic acid-containing cell-surface carbohydrates and facilitates subsequent invasion by the virus. The expression, purification, crystallization and preliminary X-ray diffraction analysis of VP8* from porcine CRW-8 rotavirus is reported. Diffraction data have been collected to 2.3 Å resolution, enabling the determination of the VP8* structure by molecular replacement.

Recombinant monovalent llama-derived antibody fragments (VHH) to rotavirus VP6 protect neonatal gnotobiotic piglets against human rotavirus-induced diarrhea.

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Vega, Celina G; Bok, Marina; Vlasova, Anastasia N; Chattha, Kuldeep S; Gómez-Sebastián, Silvia; Nuñez, Carmen; Alvarado, Carmen; Lasa, Rodrigo; Escribano, José M; Garaicoechea, Lorena L; Fernandez, Fernando; Bok, Karin; Wigdorovitz, Andrés; Saif, Linda J; Parreño, Viviana

2013-01-01

Group A Rotavirus (RVA) is the leading cause of severe diarrhea in children. The aims of the present study were to determine the neutralizing activity of VP6-specific llama-derived single domain nanoantibodies (VHH nanoAbs) against different RVA strains in vitro and to evaluate the ability of G6P[1] VP6-specific llama-derived single domain nanoantibodies (VHH) to protect against human rotavirus in gnotobiotic (Gn) piglets experimentally inoculated with virulent Wa G1P[8] rotavirus. Supplementation of the daily milk diet with 3B2 VHH clone produced using a baculovirus vector expression system (final ELISA antibody -Ab- titer of 4096; virus neutralization -VN- titer of 256) for 9 days conferred full protection against rotavirus associated diarrhea and significantly reduced virus shedding. The administration of comparable levels of porcine IgG Abs only protected 4 out of 6 of the animals from human RVA diarrhea but significantly reduced virus shedding. In contrast, G6P[1]-VP6 rotavirus-specific IgY Abs purified from eggs of hyperimmunized hens failed to protect piglets against human RVA-induced diarrhea or virus shedding when administering similar quantities of Abs. The oral administration of VHH nanoAb neither interfered with the host's isotype profiles of the Ab secreting cell responses to rotavirus, nor induced detectable host Ab responses to the treatment in serum or intestinal contents. This study shows that the oral administration of rotavirus VP6-VHH nanoAb is a broadly reactive and effective treatment against rotavirus-induced diarrhea in neonatal pigs. Our findings highlight the potential value of a broad neutralizing VP6-specific VHH nanoAb as a treatment that can complement or be used as an alternative to the current strain-specific RVA vaccines. Nanobodies could also be scaled-up to develop pediatric medication or functional food like infant milk formulas that might help treat RVA diarrhea.

Recombinant monovalent llama-derived antibody fragments (VHH to rotavirus VP6 protect neonatal gnotobiotic piglets against human rotavirus-induced diarrhea.

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Celina G Vega

Full Text Available Group A Rotavirus (RVA is the leading cause of severe diarrhea in children. The aims of the present study were to determine the neutralizing activity of VP6-specific llama-derived single domain nanoantibodies (VHH nanoAbs against different RVA strains in vitro and to evaluate the ability of G6P[1] VP6-specific llama-derived single domain nanoantibodies (VHH to protect against human rotavirus in gnotobiotic (Gn piglets experimentally inoculated with virulent Wa G1P[8] rotavirus. Supplementation of the daily milk diet with 3B2 VHH clone produced using a baculovirus vector expression system (final ELISA antibody -Ab- titer of 4096; virus neutralization -VN- titer of 256 for 9 days conferred full protection against rotavirus associated diarrhea and significantly reduced virus shedding. The administration of comparable levels of porcine IgG Abs only protected 4 out of 6 of the animals from human RVA diarrhea but significantly reduced virus shedding. In contrast, G6P[1]-VP6 rotavirus-specific IgY Abs purified from eggs of hyperimmunized hens failed to protect piglets against human RVA-induced diarrhea or virus shedding when administering similar quantities of Abs. The oral administration of VHH nanoAb neither interfered with the host's isotype profiles of the Ab secreting cell responses to rotavirus, nor induced detectable host Ab responses to the treatment in serum or intestinal contents. This study shows that the oral administration of rotavirus VP6-VHH nanoAb is a broadly reactive and effective treatment against rotavirus-induced diarrhea in neonatal pigs. Our findings highlight the potential value of a broad neutralizing VP6-specific VHH nanoAb as a treatment that can complement or be used as an alternative to the current strain-specific RVA vaccines. Nanobodies could also be scaled-up to develop pediatric medication or functional food like infant milk formulas that might help treat RVA diarrhea.

Optimizing CT technique to reduce radiation dose: effect of changes in kVp, iterative reconstruction, and noise index on dose and noise in a human cadaver.

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Chang, Kevin J; Collins, Scott; Li, Baojun; Mayo-Smith, William W

2017-06-01

For assessment of the effect of varying the peak kilovoltage (kVp), the adaptive statistical iterative reconstruction technique (ASiR), and automatic dose modulation on radiation dose and image noise in a human cadaver, a cadaver torso underwent CT scanning at 80, 100, 120 and 140 kVp, each at ASiR settings of 0, 30 and 50 %, and noise indices (NIs) of 5.5, 11 and 22. The volume CT dose index (CTDI vol ), image noise, and attenuation values of liver and fat were analyzed for 20 data sets. Size-specific dose estimates (SSDEs) and liver-to-fat contrast-to-noise ratios (CNRs) were calculated. Values for different combinations of kVp, ASiR, and NI were compared. The CTDI vol varied by a power of 2 with kVp values between 80 and 140 without ASiR. Increasing ASiR levels allowed a larger decrease in CTDI vol and SSDE at higher kVp than at lower kVp while image noise was held constant. In addition, CTDI vol and SSDE decreased with increasing NI at each kVp, but the decrease was greater at higher kVp than at lower kVp. Image noise increased with decreasing kVp despite a fixed NI; however, this noise could be offset with the use of ASiR. The CT number of the liver remained unchanged whereas that of fat decreased as the kVp decreased. Image noise and dose vary in a complicated manner when the kVp, ASiR, and NI are varied in a human cadaver. Optimization of CT protocols will require balancing of the effects of each of these parameters to maximize image quality while minimizing dose.

The Ebola virus VP35 protein is a suppressor of RNA silencing.

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Joost Haasnoot

2007-06-01

Full Text Available RNA silencing or interference (RNAi is a gene regulation mechanism in eukaryotes that controls cell differentiation and developmental processes via expression of microRNAs. RNAi also serves as an innate antiviral defence response in plants, nematodes, and insects. This antiviral response is triggered by virus-specific double-stranded RNA molecules (dsRNAs that are produced during infection. To overcome antiviral RNAi responses, many plant and insect viruses encode RNA silencing suppressors (RSSs that enable them to replicate at higher titers. Recently, several human viruses were shown to encode RSSs, suggesting that RNAi also serves as an innate defence response in mammals. Here, we demonstrate that the Ebola virus VP35 protein is a suppressor of RNAi in mammalian cells and that its RSS activity is functionally equivalent to that of the HIV-1 Tat protein. We show that VP35 can replace HIV-1 Tat and thereby support the replication of a Tat-minus HIV-1 variant. The VP35 dsRNA-binding domain is required for this RSS activity. Vaccinia virus E3L protein and influenza A virus NS1 protein are also capable of replacing the HIV-1 Tat RSS function. These findings support the hypothesis that RNAi is part of the innate antiviral response in mammalian cells. Moreover, the results indicate that RSSs play a critical role in mammalian virus replication.

A Conserved Epitope Mapped with a Monoclonal Antibody against the VP3 Protein of Goose Parvovirus by Using Peptide Screening and Phage Display Approaches.

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Li, Chenxi; Liu, Hongyu; Li, Jinzhe; Liu, Dafei; Meng, Runze; Zhang, Qingshan; Shaozhou, Wulin; Bai, Xiaofei; Zhang, Tingting; Liu, Ming; Zhang, Yun

2016-01-01

Waterfowl parvovirus (WPV) infection causes high mortality and morbidity in both geese (Anser anser) and Muscovy ducks (Cairina moschata), resulting in significant losses to the waterfowl industries. The VP3 protein of WPV is a major structural protein that induces neutralizing antibodies in the waterfowl. However, B-cell epitopes on the VP3 protein of WPV have not been characterized. To understand the antigenic determinants of the VP3 protein, we used the monoclonal antibody (mAb) 4A6 to screen a set of eight partially expressed overlapping peptides spanning VP3. Using western blotting and an enzyme-linked immunosorbent assay (ELISA), we localized the VP3 epitope between amino acids (aa) 57 and 112. To identify the essential epitope residues, a phage library displaying 12-mer random peptides was screened with mAb 4A6. Phage clone peptides displayed a consensus sequence of YxRFHxH that mimicked the sequence 82Y/FNRFHCH88, which corresponded to amino acid residues 82 to 88 of VP3 protein of WPVs. mAb 4A6 binding to biotinylated fragments corresponding to amino acid residues 82 to 88 of the VP3 protein verified that the 82FxRFHxH88 was the VP3 epitope and that amino acids 82F is necessary to retain maximal binding to mAb 4A6. Parvovirus-positive goose and duck sera reacted with the epitope peptide by dot blotting assay, revealing the importance of these amino acids of the epitope in antibody-epitope binding reactivity. We identified the motif FxRFHxH as a VP3-specific B-cell epitope that is recognized by the neutralizing mAb 4A6. This finding might be valuable in understanding of the antigenic topology of VP3 of WPV.

Expression of VP60 gene from rabbit haemorrhagic disease virus ...

African Journals Online (AJOL)

The VP60 gene from rabbit haemorrhagic disease virus (RHDV) YL strain in Northeast of China, under control of the ats1A promoter from Rubisco small subunit genes of Arabidopsis thaliana, was introduced into the transfer deoxyribonucleic acid (T-DNA) region of plant transfer vector pCAMBIA1300 and transferred to ...

Relationship between apoptosis and the BH2 domain sequence of the VP5 peptide of infectious pancreatic necrosis virus

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Cesar Ortega S.

2014-03-01

Full Text Available Objective. To determine whether the level of apoptosis induced by infectious pancreatic necrosis virus (IPNV is related to the amino acid sequence of the BH2 domain of the VP5 protein and the level of infectivity. Materials and methods. Three IPNV strains were used, the VP2 protein gene was amplified for genotyping and the VP5 sequence was also obtained. The infectivity of the strains was calculated using the viral titer obtained at 12, 24, 36 and 45 hpi in CHSE-214 cells. The percentage of apoptosis in infected cells was visualized by TUNEL assay and immunohistochemistry (caspase 3 detection. Results. The V70/06 and V33/98 strains corresponded to genotype Sp, while V112/06 to VR-299; the amino acid analysis of the V70/06 strain allows its classification as middle virulent strain and V33/98 and V112/06 strains as low virulent ones; infection with the V112/06 strain produced a lower viral titer (p0.05. Conclusions. The results showed that the differences in the BH2 sequence of the VP5 protein, infectivity and the VP2 sequence are not associated with the modulation of apoptosis.

Measurement of kVp, PPV and air kerma values in function of the electric current quantity and the focus-detector distance in one X-ray equipment

International Nuclear Information System (INIS)

Lucena, Rodrigo F. de; Potiens, Maria da Penha A.; Vivolo, Vitor

2009-01-01

The objective of this work was to study the behavior of the X-ray equipment Pantak/Seifert, model MXR-160/22 of the calibration laboratory of IPEN, LCI, operating in the diagnostic radiology radiation quality RQR 5 (70 kV). For this evaluation it was used a noninvasive meter PTW, Diavolt TM model. The measurements of kVp, PPV and Dose (air kerma), were made varying the electric current and distance between the focal point and the meter. This behavior is described in the literature and was expected in the analysis of the measurements for comparison purposes. For the tests where it was only increased the electric current it was waited a linear increase of the dose (air kerma), but not a variation in the kVp and PPV. The measurements had corresponded to the waited behavior, since the Dose (air kerma) measurements presented a linear increase with the increase of the electric current and the kVp and PPV values showed a variation less than 2%. In the corresponding measurements increasing the distance between focal point and meter, it was waited the exponentially decreasing of the Dose (air kerma) and again a small variation or no variation of the PPV and kVP with the increase of the distance. Over again the measurements corresponded to the expected, where the Dose (air kerma) decreased exponentially and the PPV and the kVp had a variation less than 1.5%. (author)

High yield expression in a recombinant E. coli of a codon optimized chicken anemia virus capsid protein VP1 useful for vaccine development

Directory of Open Access Journals (Sweden)

You Bang-Jau

2011-07-01

Full Text Available Abstract Background Chicken anemia virus (CAV, the causative agent chicken anemia, is the only member of the genus Gyrovirus of the Circoviridae family. CAV is an immune suppressive virus and causes anemia, lymph organ atrophy and immunodeficiency. The production and biochemical characterization of VP1 protein and its use in a subunit vaccine or as part of a diagnostic kit would be useful to CAV infection prevention. Results Significantly increased expression of the recombinant full-length VP1 capsid protein from chicken anemia virus was demonstrated using an E. coli expression system. The VP1 gene was cloned into various different expression vectors and then these were expressed in a number of different E. coli strains. The expression of CAV VP1 in E. coli was significantly increased when VP1 was fused with GST protein rather than a His-tag. By optimizing the various rare amino acid codons within the N-terminus of the VP1 protein, the expression level of the VP1 protein in E. coli BL21(DE3-pLysS was further increased significantly. The highest protein expression level obtained was 17.5 g/L per liter of bacterial culture after induction with 0.1 mM IPTG for 2 h. After purification by GST affinity chromatography, the purified full-length VP1 protein produced in this way was demonstrated to have good antigenicity and was able to be recognized by CAV-positive chicken serum in an ELISA assay. Conclusions Purified recombinant VP1 protein with the gene's codons optimized in the N-terminal region has potential as chimeric protein that, when expressed in E. coli, may be useful in the future for the development of subunit vaccines and diagnostic tests.

Raltegravir versus lopinavir/ritonavir for treatment of HIV-infected late-presenting pregnant women.

Science.gov (United States)

Brites, Carlos; Nóbrega, Isabella; Luz, Estela; Travassos, Ana Gabriela; Lorenzo, Cynthia; Netto, Eduardo M

2018-06-01

Background Late-presenting pregnant women pose a challenge in the prevention of HIV-1 mother-to-child-transmission. We compared the safety and efficacy of raltegravir and lopinavir/ritonavir for this population. Methods We did a single-center, pilot, open-label, randomized trial in Brazil (N = 44). We randomly allocated late-presenting HIV-infected pregnant women (older than 18 years with a plasma HIV-1 RNA >1000 copies/mL) to receive raltegravir 400 mg twice a day or lopinavir/ritonavir 400/100 mg twice a day plus zidovudine and lamivudine (1:1). The primary endpoint was virological suppression at delivery (HIV-1 RNA HIV-infected late-presenting pregnant women.

Neonatal treatment philosophy in Dutch and German NICUs: Health-Related Quality of Life in Adulthood of VP/VLBW infants

NARCIS (Netherlands)

Breeman, L.D.|info:eu-repo/dai/nl/390776114; van der Pal, Sylvia; Verrips, Gijsbert; Baumann, Nicole; Bartmann, Peter; Wolke, Dieter

2017-01-01

Purpose. Although survival after very preterm birth (VP) / very low birth weight (VLBW) has improved, a significant number of VP/VLBW individuals develop physical and cognitive problems during their life course that may affect their health-related quality of life (HRQoL). We compared HRQoL in

The Kinase STK3 Interacts with the Viral Structural Protein VP1 and Inhibits Foot-and-Mouth Disease Virus Replication

Science.gov (United States)

Xue, Qiao

2017-01-01

Foot-and-mouth disease virus (FMDV) is the etiological agent of FMD, which affects domestic and wild cloven-hoofed animals. The structural protein VP1 plays an important role in FMDV pathogenesis. However, the interacting partners of VP1 in host cells and the effects of these interactions in FMDV replication remain incompletely elucidated. Here, we identified a porcine cell protein, serine/threonine kinase 3 (STK3), which interacts with FMDV VP1 using the yeast two-hybrid system. The VP1-STK3 interaction was further confirmed by coimmunoprecipitation experiments in human embryonic kidney 293T and porcine kidney 15 (PK-15) cells. The carboxyl-terminal region (amino acids 180–214) of VP1 was essential for its interaction with STK3. The effects of overexpression and underexpressing of STK3 in PK-15 cells were assessed, and the results indicated that STK3 significantly inhibited FMDV replication. Our data expand the role of STK3 during viral infection, provide new information regarding the host cell kinases that are involved in viral replication, and identify potential targets for future antiviral strategies. PMID:29226127

The color tuning of PS-b-P2VP lamellar films with changing the alkyl chain length of 1-iodoalkanes.

Science.gov (United States)

Shin, Sung-Eui; Kim, Su-Young; Shin, Dong-Myung

2011-05-01

Photonic crystals with tunability in the visible or near-infrared region have drawn increasing attention for controlling and processing light for the active components of future display. We prepared polystyrene-b-poly(2-vinyl pyridine) (PS-b-P2VP) lamellar films which is hydrophobic block-hydrophilic polyelectrolyte block polymer of 57 kg/mol-b-57 kg/mol. The lamellar stacks, which is alternating layer of hydrophilic and hydrophobic moiety of PS-b-P2VP, are obtained by exposing the spin coated film under chloroform vapor. The band gaps of the lamellar films interestingly varied after immersion into the quaternizing solvents containing 5 wt% of iodomethane, iodoethane, 1-iodobutane, 1-iodopentane, 1-iodohexane and 1-iodooctane solubilized in n-hexane. The iodoalkanes reacted with pyridine groups in PS-b-P2VP and generated the alkyl pyridinium salts readily. The degree of quaternization, alkyl chain length of iodoalkane and the salt water concentration affects the spacing of layer structure of PS-b-P2VP. The iodomethane and iodohexane produced similar band gaps and salt concentration dependence. These results are very much dependent on the hydrophobic-hydrophilic characters of PS-b-P2VP lamellar surface.

System and method for investigating sub-surface features and 3D imaging of non-linear property, compressional velocity VP, shear velocity VS and velocity ratio VP/VS of a rock formation

Science.gov (United States)

Vu, Cung Khac; Skelt, Christopher; Nihei, Kurt; Johnson, Paul A.; Guyer, Robert; Ten Cate, James A.; Le Bas, Pierre-Yves; Larmat, Carene S.

2015-06-02

A system and a method for generating a three-dimensional image of a rock formation, compressional velocity VP, shear velocity VS and velocity ratio VP/VS of a rock formation are provided. A first acoustic signal includes a first plurality of pulses. A second acoustic signal from a second source includes a second plurality of pulses. A detected signal returning to the borehole includes a signal generated by a non-linear mixing process from the first and second acoustic signals in a non-linear mixing zone within an intersection volume. The received signal is processed to extract the signal over noise and/or signals resulting from linear interaction and the three dimensional image of is generated.

Antigenic profile of African horse sickness virus serotype 4 VP5 and identification of a neutralizing epitope shared with bluetongue virus and epizootic hemorrhagic disease virus

DEFF Research Database (Denmark)

Martinez-Torrecuadrada, J.L.; Langeveld, J.P.M.; Venteo, A.

1999-01-01

African horse sickness virus (AHSV) causes a fatal disease in horses. The virus capsid is composed of a double protein layer, the outermost of which is formed by two proteins: VP2 and VP5. VP2 is known to determine the serotype of the virus and to contain the neutralizing epitopes. The biological...... in a plaque reduction assay were generated. To dissect the antigenic structure of AHSV VP5, the protein was cloned in Escherichia coil using the pET3 system. The immunoreactivity of both MAbs, and horse and rabbit polyclonal antisera, with 17 overlapping fragments from VP5 was analyzed. The most....... Neutralizing epitopes were defined at positions 85-92 (PDPLSPGE) for MAb 10AE12 and at 179-185 (EEDLRTR) for MAb 10AC6. Epitope 10AE12 is highly conserved between the different orbiviruses. MAb 10AE12 was able to recognize bluetongue virus VP5 and epizootic hemorrhagic disease virus VP5 by several techniques...

Elymus dahuricus H+-PPase EdVP1 enhances potassium uptake and utilization of wheat through the development of root system

OpenAIRE

Ruan, L; Zhang, J; Xin, X; Miller, A. J; Tong, Y

2013-01-01

We investigated the differences of K acquisition and utilization, morphological and physiological characteristics of roots and grain yield between Elymus dahuricus H+-PPase (EdVP1) transgenic wheat and wild type wheat under low K stress. The results showed that, the grain yield and K economic utilization index (KUI-E) in wild type wheat were only 61.14% and 50.20% of those in EdVP1 transgenic wheat. EdVP1 increased the free IAA accumulations in roots, which may play a key role in the developm...

CT angiography of the aorta using 80 kVp in combination with sinogram-affirmed iterative reconstruction and automated tube current modulation: Effects on image quality and radiation dose

International Nuclear Information System (INIS)

Boos, Johannes; Aissa, Joel; Lanzman, Rotem S.; Heusch, Philipp; Schimmoller, Lars; Schleich, Christoph; Thomas, Christoph; Antoch, Gerald; Kropil, Patric

2016-01-01

The objective of this study was to evaluate image quality and radiation dose of a CT angiography (CTA) protocol using 80 kVp in combination with iterative reconstruction and automated tube current modulation. Ninety-five aortic CTA examinations were included in this study. A novel 80 kVp aortic CTA-protocol with iterative reconstruction was introduced in our department in March 2012 for patients with a body mass index (BMI) below 32 kg/m 2 . The first 72 consecutive examinations were retrospectively assigned to group A (56 patients, 42 men, 14 women, mean age 69.6 ± 10.7years, BMI range 19.7–31.1 kg/m 2 ). For comparison, the last 23 consecutive examinations performed with the old protocol (100 kVp) were assigned to group B (21 patients, 13 men, 8 women, mean age 67.4 ± 11.1years, BMI range 19.7–31.9 kg/m 2 ). Thoracic and abdominal contrast-to-noise ratio (CNR), signal-to-noise ratio (SNR) and aortic attenuation were assessed. Subjective image quality was rated on a 5-point scale (1 = non diagnostic; 5 = excellent). Furthermore, dose length product (DLP) and volumetric computed tomography dose index (CTDI vol ) were analysed. All examinations achieved diagnostic image quality. Attenuation of the aorta was significantly higher in group A compared with B (thoracic: 443.5 ± 90.5 Hounsfield units (HU) vs. 296.0 ± 61.0 HU; abdominal: 426.3 ± 94.2 HU vs. 283.6 ± 60.5 HU; P < 0.05, respectively). CNR, SNR and subjective image quality were comparable between both groups (CNR: 12.8 ± 3.7 vs. 13.0 ± 7.4; SNR 14.4 ± 3.9 vs. 14.9 ± 8.2; subjective image quality: 4.3 ± 0.6 vs. 4.5 ± 0.6; P > 0.05, respectively). CTDI vol and DLP were significantly lower in group A (1.9 ± 0.5 mGy; 139.2 ± 41.1 mGy × cm) as compared with group B (4.2 ± 1.4 mGy; 292.1 ± 91.5 mGy × cm; P < 0.001, respectively). Low-dose CTA of the aorta using 80 kVp with iterative

The influence of film-screen color sensitivity and type of measurement device on kVp measurements.

Science.gov (United States)

Lam, R W; Price, S C

1989-01-01

Three methods for evaluating radiographic kVp were studied: the Wisconsin Test Cassette, the Noninvasive Evaluator of Radiation Outputs (NERO), and the Dynalyzer. The Dynalyzer kVp readings were the highest and were followed by NERO and cassette readings in descending order. By film type, the cassette readings ranged from Kodak OG (green sensitive), TMG (green sensitive), XK (blue sensitive), and XRP (blue sensitive) in descending order. The results show that there is significant variation between the methods.

Minimizing Contrast Medium Doses to Diagnose Pulmonary Embolism with 80-kVp Multidetector Computed Tomography in Azotemic Patients

Energy Technology Data Exchange (ETDEWEB)

Holmquist, F. (Dept. of Diagnostic Radiology, Malmoe Univ. Hospital, Univ. of Lund, Malmoe (Sweden)); Hansson, K.; Pasquariello, F. (Dept. of Internal Medicine, Lasarettet Trelleborg, Univ. of Lund, Trelleborg (Sweden)); Bjoerk, J. (Competence Center for Clinical Research, Univ. Hospital, Univ. of Lund, Lund (Sweden)); Nyman, U. (Dept. of Radiology, Lasarettet Trelleborg, Univ. of Lund, Trelleborg (Sweden))

2009-02-15

Background: In diagnosing acute pulmonary embolism (PE) in azotemic patients, scintigraphy and magnetic resonance imaging are frequently inconclusive or not available in many hospitals. Computed tomography is readily available, but relatively high doses (30-50 g I) of potentially nephrotoxic iodine contrast media (CM) are used. Purpose: To report on the diagnostic quality and possible contrast-induced nephropathy (CIN) after substantially reduced CM doses to diagnose PE in azotemic patients using 80-peak kilovoltage (kVp) 16-row multidetector computed tomography (MDCT) combined with CM doses tailored to body weight, fixed injection duration adapted to scan time, automatic bolus tracking, and saline chaser. Material and Methods: Patients with estimated glomerular filtration rate (eGFR) <50 ml/min were scheduled to undergo 80-kVp MDCT using 200 mg I/kg, and those with eGFR =50 ml/min, 120-kVp MDCT with 320 mg I/kg. Both protocols used an 80-kg maximum dose weight and a fixed 15-s injection time. Pulmonary artery density and contrast-to-noise ratio were measured assuming 70 Hounsfield units (HU) for a fresh clot. CIN was defined as a plasma creatinine rise >44.2 mumol/l from baseline. Results: 89/148 patients (63/68 females) underwent 80-/120-kVp protocols, respectively, with 95% of the examinations being subjectively excellent or adequate. Mean values in the 80-/120-kVp cohorts regarding age were 82/65 years, body weight 66/78 kg, effective mAs 277/117, CM dose 13/23 g I, pulmonary artery density 359/345 HU, image noise (1 standard deviation) 24/21 HU, contrast-to-noise ratio 13/13, and dose-length product 173/258 mGycm. Only 1/65 and 2/119 patients in the 80- and 120-kVp cohorts, respectively, with negative CT and no anticoagulation suffered non-fatal thromboembolism during 3-month follow-up. No patient developed CIN. Conclusion: 80-kVp 16-row MDCT with optimization of injection parameters may be performed with preserved diagnostic quality, using markedly reduced CM

10 kVp rule – An anthropomorphic pelvis phantom imaging study using a CR system: Impact on image quality and effective dose using AEC and manual mode

International Nuclear Information System (INIS)

Lança, Luís; Franco, Loris; Ahmed, Abdulfatah; Harderwijk, Marloes; Marti, Chloe; Nasir, Sadeeda; Ndlovu, Junior; Oliveira, Miguel; Santiago, Ana Rita; Hogg, Peter

2014-01-01

Purpose: This study aims to investigate the influence of tube potential (kVp) variation in relation to perceptual image quality and effective dose (E) for pelvis using automatic exposure control (AEC) and non-AEC in a Computed Radiography (CR) system. Methods and materials: To determine the effects of using AEC and non-AEC by applying the 10 kVp rule in two experiments using an anthropomorphic pelvis phantom. Images were acquired using 10 kVp increments (60–120 kVp) for both experiments. The first experiment, based on seven AEC combinations, produced 49 images. The mean mAs from each kVp increment were used as a baseline for the second experiment producing 35 images. A total of 84 images were produced and a panel of 5 experienced observers participated for the image scoring using the two alternative forced choice (2AFC) visual grading software. PCXMC software was used to estimate E. Results: A decrease in perceptual image quality as the kVp increases was observed both in non-AEC and AEC experiments, however no significant statistical differences (p > 0.05) were found. Image quality scores from all observers at 10 kVp increments for all mAs values using non-AEC mode demonstrates a better score up to 90 kVp. E results show a statistically significant decrease (p = 0.000) on the 75th quartile from 0.37 mSv at 60 kVp to 0.13 mSv at 120 kVp when applying the 10 kVp rule in non-AEC mode. Conclusion: Using the 10 kVp rule, no significant reduction in perceptual image quality is observed when increasing kVp whilst a marked and significant E reduction is observed

Cerebral bone subtraction CT angiography using 80 kVp and sinogram-affirmed iterative reconstruction: contrast medium and radiation dose reduction with improvement of image quality

Energy Technology Data Exchange (ETDEWEB)

Nagayama, Yasunori [Kumamoto City Hospital, Department of Radiology, Kumamoto (Japan); Kumamoto University, Department of Diagnostic Radiology, Chuo-ku, Kumamoto (Japan); Nakaura, Takeshi; Oda, Seitaro; Kidoh, Masafumi; Utsunomiya, Daisuke; Yamashita, Yasuyuki [Kumamoto University, Department of Diagnostic Radiology, Chuo-ku, Kumamoto (Japan); Tsuji, Akinori; Urata, Joji; Furusawa, Mitsuhiro; Yuki, Hideaki; Hirarta, Kenichiro [Kumamoto City Hospital, Department of Radiology, Kumamoto (Japan)

2017-02-15

The purpose of this study was to evaluate the feasibility of a contrast medium (CM), radiation dose reduction protocol for cerebral bone-subtraction CT angiography (BSCTA) using 80-kVp and sinogram-affirmed iterative reconstruction (SAFIRE). Seventy-five patients who had undergone BSCTA under the 120- (n = 37) or the 80-kVp protocol (n = 38) were included. CM was 370 mgI/kg for the 120-kVp and 296 mgI/kg for the 80-kVp protocol; the 120- and the 80-kVp images were reconstructed with filtered back-projection (FBP) and SAFIRE, respectively. We compared effective dose (ED), CT attenuation, image noise, and contrast-to-noise ratio (CNR) of two protocols. We also scored arterial contrast, sharpness, depiction of small arteries, visibility near skull base/clip, and overall image quality on a four-point scale. ED was 62% lower at 80- than 120-kVp (0.59 ± 0.06 vs 1.56 ± 0.13 mSv, p < 0.01). CT attenuation of the internal carotid artery (ICA) and middle cerebral artery (MCA) was significantly higher on 80- than 120-kVp (ICA: 557.4 ± 105.7 vs 370.0 ± 59.3 Hounsfield units (HU), p < 0.01; MCA: 551.9 ± 107.9 vs 364.6 ± 62.2 HU, p < 0.01). The CNR was also significantly higher on 80- than 120-kVp (ICA: 46.2 ± 10.2 vs 36.9 ± 7.6, p < 0.01; MCA: 45.7 ± 10.0 vs 35.7 ± 9.0, p < 0.01). Visibility near skull base and clip was not significantly different (p = 0.45). The other subjective scores were higher with the 80- than the 120-kVp protocol (p < 0.05). The 80-kVp acquisition with SAFIRE yields better image quality for BSCTA and substantial reduction in the radiation and CM dose compared to the 120-kVp with FBP protocol. (orig.)

Kobuvirus VP3 protein restricts the IFN-β-triggered signaling pathway by inhibiting STAT2-IRF9 and STAT2-STAT2 complex formation

Energy Technology Data Exchange (ETDEWEB)

Peng, Qianqian; Lan, Xi; Wang, Chen [State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046 (China); Ren, Yujie; Yue, Ningning [College of Life Sciences, Wuhan University, Wuhan 430072 (China); Wang, Junyong [State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046 (China); Zhong, Bo [College of Life Sciences, Wuhan University, Wuhan 430072 (China); Medical Research Institute, School of Medicine, Wuhan University, Wuhan 430071 (China); Zhu, Qiyun, E-mail: zhuqiyun@caas.cn [State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046 (China)

2017-07-15

Emerged porcine kobuvirus (PKV) has adversely affected the global swine industry since 2008, but the etiological biology of PKV is unclear. Screening PKV-encoded structural and non-structural proteins with a type I IFN-responsive luciferase reporter showed that PKV VP3 protein inhibited the IFN-β-triggered signaling pathway, resulting in the decrease of VSV-GFP replication. QPCR data showed that IFN-β downstream cytokine genes were suppressed without cell-type specificity as well. The results from biochemical experiments indicated that PKV VP3 associated with STAT2 and IRF9, and interfered with the formation of the STAT2-IRF9 and STAT2-STAT2 complex, impairing nuclear translocation of STAT2 and IRF9. Taken together, these data reveal a new mechanism for immune evasion of PKV. - Highlights: •PKV VP3 inhibits the IFN-β-triggered signaling pathway. •VP3 associates with STAT2 and IRF9. •VP3 blocks the STAT2-IRF9 nuclear translocation. •VP3 utilizes a novel strategy for innate immune evasion.

Increased expression of Matrix Metalloproteinase 9 in liver from NZB/W F1 mice received antibody against human parvovirus B19 VP1 unique region protein

Directory of Open Access Journals (Sweden)

Hsu Gwo-Jong

2009-01-01

Full Text Available Abstract Background Human parvovirus B19 infection has been postulated to the anti-phospholipid syndrome (APS in autoimmunity. However, the influence of anti-B19-VP1u antibody in autoimmune diseases is still obscure. Methods To elucidate the effect of anti-B19-VP1u antibodies in systemic lupus erythematosus (SLE, passive transfer of rabbit anti-B19-VP1u IgG was injected intravenously into NZB/W F1 mice. Results Significant reduction of platelet count and prolonged thrombocytopenia time were detected in anti-B19-VP1u IgG group as compared to other groups, whereas significant increases of anti-B19-VP1u, anti-phospholipid (APhL, and anti-double strand DNA (dsDNA antibody binding activity were detected in anti-B19-VP1u group. Additionally, significant increases of matrix metalloproteinase-9 (MMP9 activity and protein expression were detected in B19-VP1u IgG group. Notably, phosphatidylinositol 3-phosphate kinase (PI3K and phosphorylated extracellular signal-regulated kinase (ERK proteins were involved in the induction of MMP9. Conclusion These experimental results firstly demonstrated the aggravated effects of anti-B19-VP1u antibody in disease activity of SLE.

Phylogenetic analysis of VP2 gene of canine parvovirus and comparison with Indian and world isolates.

Science.gov (United States)

Kaur, G; Chandra, M; Dwivedi, P N

2016-03-01

Canine parvovirus (CPV) causes hemorrhagic enteritis, especially in young dogs, leading to high morbidity and mortality. It has four main antigenic types CPV-2, CPV-2a, CPV-2b and CPV-2c. Virus protein 2 (VP2) is the main capsid protein and mutations affecting VP2 gene are responsible for the evolution of various antigenic types of CPV. Full length VP2 gene from field isolates was amplified and cloned for sequence analysis. The sequences were submitted to the GenBank and were assigned Acc. Nos., viz. KP406928.1 for P12, KP406927.1 for P15, KP406930.1 for P32, KP406926.1 for Megavac-6 and KP406929.1 for NobivacDHPPi. Phylogenetic analysis indicated that the samples were forming a separate clad with vaccine strains. When the samples were compared with the world and Indian isolates, it was observed that samples formed a separate node indicating regional genetic variation in CPV.

Prevalence and Correlates of Having a Regular Physician among Women Presenting for Induced Abortion.

Science.gov (United States)

Chor, Julie; Hebert, Luciana E; Hasselbacher, Lee A; Whitaker, Amy K

2016-01-01

To determine the prevalence and correlates of having a regular physician among women presenting for induced abortion. We conducted a retrospective review of women presenting to an urban, university-based family planning clinic for abortion between January 2008 and September 2011. We conducted bivariate analyses, comparing women with and without a regular physician, and multivariable regression modeling, to identify factors associated with not having a regular physician. Of 834 women, 521 (62.5%) had a regular physician and 313 (37.5%) did not. Women with a prior pregnancy, live birth, or spontaneous abortion were more likely than women without these experiences to have a regular physician. Women with a prior induced abortion were not more likely than women who had never had a prior induced abortion to have a regular physician. Compared with women younger than 18 years, women aged 18 to 26 years were less likely to have a physician (adjusted odds ratio [aOR], 0.25; 95% confidence interval [CI], 0.10-0.62). Women with a prior live birth had increased odds of having a regular physician compared with women without a prior pregnancy (aOR, 1.89; 95% CI, 1.13-3.16). Women without medical/fetal indications and who had not been victims of sexual assault (self-indicated) were less likely to report having a regular physician compared with women with medical/fetal indications (aOR, 0.55; 95% CI, 0.17-0.82). The abortion visit is a point of contact with a large number of women without a regular physician and therefore provides an opportunity to integrate women into health care. Copyright © 2016 Jacobs Institute of Women's Health. Published by Elsevier Inc. All rights reserved.

The structure of avian polyomavirus reveals variably sized capsids, non-conserved inter-capsomere interactions, and a possible location of the minor capsid protein VP4

International Nuclear Information System (INIS)

Shen, Peter S.; Enderlein, Dirk; Nelson, Christian D.S.; Carter, Weston S.; Kawano, Masaaki; Xing Li; Swenson, Robert D.; Olson, Norman H.; Baker, Timothy S.; Cheng, R. Holland; Atwood, Walter J.; Johne, Reimar; Belnap, David M.

2011-01-01

Avian polyomavirus (APV) causes a fatal, multi-organ disease among several bird species. Using cryogenic electron microscopy and other biochemical techniques, we investigated the structure of APV and compared it to that of mammalian polyomaviruses, particularly JC polyomavirus and simian virus 40. The structure of the pentameric major capsid protein (VP1) is mostly conserved; however, APV VP1 has a unique, truncated C-terminus that eliminates an intercapsomere-connecting β-hairpin observed in other polyomaviruses. We postulate that the terminal β-hairpin locks other polyomavirus capsids in a stable conformation and that absence of the hairpin leads to the observed capsid size variation in APV. Plug-like density features were observed at the base of the VP1 pentamers, consistent with the known location of minor capsid proteins VP2 and VP3. However, the plug density is more prominent in APV and may include VP4, a minor capsid protein unique to bird polyomaviruses.

The VP3 factor from viruses of Birnaviridae family suppresses RNA silencing by binding both long and small RNA duplexes.

Science.gov (United States)

Valli, Adrian; Busnadiego, Idoia; Maliogka, Varvara; Ferrero, Diego; Castón, José R; Rodríguez, José Francisco; García, Juan Antonio

2012-01-01

RNA silencing is directly involved in antiviral defense in a wide variety of eukaryotic organisms, including plants, fungi, invertebrates, and presumably vertebrate animals. The study of RNA silencing-mediated antiviral defences in vertebrates is hampered by the overlap with other antiviral mechanisms; thus, heterologous systems are often used to study the interplay between RNA silencing and vertebrate-infecting viruses. In this report we show that the VP3 protein of the avian birnavirus Infectious bursal disease virus (IBDV) displays, in addition to its capacity to bind long double-stranded RNA, the ability to interact with double-stranded small RNA molecules. We also demonstrate that IBDV VP3 prevents the silencing mediated degradation of a reporter mRNA, and that this silencing suppression activity depends on its RNA binding ability. Furthermore, we find that the anti-silencing activity of IBDV VP3 is shared with the homologous proteins expressed by both insect- and fish-infecting birnaviruses. Finally, we show that IBDV VP3 can functionally replace the well-characterized HCPro silencing suppressor of Plum pox virus, a potyvirus that is unable to infect plants in the absence of an active silencing suppressor. Altogether, our results support the idea that VP3 protects the viral genome from host sentinels, including those of the RNA silencing machinery.

Identification of canine parvovirus with the Q370R point mutation in the VP2 gene from a giant panda (Ailuropoda melanoleuca)

Science.gov (United States)

2013-01-01

Background In this study, we sequenced and phylogenetic analyses of the VP2 genes from twelve canine parvovirus (CPV) strains obtained from eleven domestic dogs and a giant panda (Ailuropoda melanoleuca) in China. A novel canine parvovirus (CPV) was detected from the giant panda in China. Results Nucleotide and phylogenetic analysis of the capsid protein VP2 gene classified the CPV as a new CPV-2a type. Substitution of Gln for Arg at the conserved 370 residue in CPV presents an unusual variation in the new CPV-2a amino acid sequence of the giant panda and is further evidence for the continuing evolution of the virus. Conclusions These findings extend the knowledge on CPV molecular epidemiology of particular relevance to wild carnivores. PMID:23706032

Charmless B→VP decays using flavor SU(3) symmetry

International Nuclear Information System (INIS)

Chiang Chengwei; Gronau, Michael; Luo Zumin; Rosner, Jonathan L.; Suprun, Denis A.

2004-01-01

The decays of B mesons to a charmless vector (V) and pseudoscalar (P) meson are analyzed within a framework of flavor SU(3) in which symmetry breaking is taken into account through ratios of decay constants in tree (T) amplitudes but exact SU(3) is assumed for color-suppressed and penguin amplitudes. The magnitudes and relative phases of tree and penguin amplitudes are extracted from data, the symmetry assumption is tested, and predictions are made for rates and CP asymmetries in as-yet-unseen decay modes. A key assumption for which we perform some tests and suggest others is a relation between penguin amplitudes in which the spectator quark is incorporated into either a pseudoscalar meson or a vector meson. Values of γ slightly restricting the range currently allowed by fits to other data are favored, but outside this range there remain acceptable solutions which cannot be excluded solely on the basis of present B→VP experiments

An Examination of Women's Self-Presentation, Social Physique Anxiety, and Setting Preferences during Injury Rehabilitation

Science.gov (United States)

Hall, Craig R.

2017-01-01

Objectives. This study investigated whether women experience self-presentational concerns related to rehabilitation settings and explored preferences for characteristics of the social and physical treatment environment in relation to women's Social Physique Anxiety (SPA). Methods. Two cross-sectional studies were conducted. In Study 1, female undergraduate students (n = 134) completed four questionnaires (Social Physique Anxiety Scale; three bespoke questionnaires assessing self-presentation in rehabilitation and social and physical environment preferences) with respect to hypothetical rehabilitation scenarios. Study 2 recruited injured women who were referred for physiotherapy (n = 62) to complete the same questionnaires regarding genuine rehabilitation scenarios. Results. Women with high SPA showed less preference for physique salient clothing than women with low SPA in both hypothetical (p = 0.001) and genuine settings (p = 0.01). In Study 2, women with high SPA also preferred that others in the clinic were female (p = 0.01) and reported significantly greater preference for private treatment spaces (p = 0.05). Conclusions. Self-presentational concerns exist in rehabilitation as in exercise settings. Results indicated inverse relationships between women's SPA and preference for the presence of men, physique-enhancing clothing, and open-concept treatment settings. Future studies to determine the effect of self-presentational concerns on treatment adherence are needed. PMID:28386484

Breast cancer awareness and barriers to symptomatic presentation among women from different ethnic groups in East London

Science.gov (United States)

Forbes, L J L; Atkins, L; Thurnham, A; Layburn, J; Haste, F; Ramirez, A J

2011-01-01

Background: During 2001 to 2005, 1-year breast cancer survival was low in ethnically diverse East London. We hypothesised that this was due to low breast cancer awareness and barriers to symptomatic presentation, leading to late stage at diagnosis in women from ethnic minorities. We examined ethnic differences in breast cancer awareness and barriers to symptomatic presentation in East London. Methods: We carried out a population-based survey of 1515 women aged 30+ using the Cancer Research UK Breast Cancer Awareness Measure. We analysed the data using logistic regression adjusting for age group and level of deprivation. Results: South Asian and black women had lower breast cancer awareness than white women. South Asian women, but not black women, reported more emotional barriers to seeking medical help than white women. White women were more likely than non-white women to report worry about wasting the doctor's time as a barrier to symptomatic presentation. Conclusion: Interventions to promote early presentation of breast cancer for South Asian and black women should promote knowledge of symptoms and skills to detect changes, and tackle emotional barriers to symptomatic presentation and for white women tackle the idea that going to the doctor to discuss a breast symptom will waste the doctor's time. PMID:21989188

Co-expression of apoptin (VP3) and antibacterial peptide cecropin B ...

African Journals Online (AJOL)

The antibacterial peptide cecropin B mutant (ABPS1) gene has a broad range of antibacterial and antiproliferative properties. Apoptin (VP3), a chicken anaemia virus-encoded protein is known to induce apoptosis in human transformed cells. To explore drug combination in human tumor cells, apoptin and ABPS1 eukaryotic ...

Expression of the VP40 antigen from the Zaire ebolavirus in tobacco plants.

Science.gov (United States)

Monreal-Escalante, Elizabeth; Ramos-Vega, Abel A; Salazar-González, Jorge A; Bañuelos-Hernández, Bernardo; Angulo, Carlos; Rosales-Mendoza, Sergio

2017-07-01

The plant cell is able to produce the VP40 antigen from the Zaire ebolavirus , retaining the antigenicity and the ability to induce immune responses in BALB/c mice. The recent Ebola outbreak evidenced the need for having vaccines approved for human use. Herein we report the expression of the VP40 antigen from the Ebola virus as an initial effort in the development of a plant-made vaccine that could offer the advantages of being cheap and scalable, which is proposed to overcome the rapid need for having vaccines to deal with future outbreaks. Tobacco plants were transformed by stable DNA integration into the nuclear genome using the CaMV35S promoter and a signal peptide to access the endoplasmic reticulum, reaching accumulation levels up to 2.6 µg g -1 FW leaf tissues. The antigenicity of the plant-made VP40 antigen was evidenced by Western blot and an initial immunogenicity assessment in test animals that revealed the induction of immune responses in BALB/c mice following three weekly oral or subcutaneous immunizations at very low doses (125 and 25 ng, respectively) without accessory adjuvants. Therefore, this plant-based vaccination prototype is proposed as an attractive platform for the production of vaccines in the fight against Ebola virus disease outbreaks.

In Situ GISAXS Study on Solvent Vapour Induced Orientation Switching in PS-b-P4VP Block Copolymer Thin Films

International Nuclear Information System (INIS)

Gowd, E Bhoje; Boehme, Marcus; Stamm, Manfred

2010-01-01

We investigated the orientation changes of cylindrical P4VP microdomains in PS-b-P4VP thin films upon annealing in different solvent vapours using the time-resolved in situ grazing-incidence small-angle X-ray scattering (GISAXS) for the first time. Swelling of perpendicular cylinders (C perpendicular) in a non-selective solvent vapours (chloroform) leads to the orientation change to in-plane cylinders (C//) and it occurs through a disordered state. On the other hand, swelling of perpendicular cylinders (C perpendicular) in a selective solvent vapours (1,4-dioxane) leads the morphological change from cylindrical to BCC spherical morphology. Solvent evaporation results in shrinkage of the matrix in the vertical direction and subsequently merges the spheres into the perpendicularly aligned cylinders. The selectivity of the solvent to constituting blocks and the solvent evaporation rate may be mainly responsible for such orientation change of cylindrical P4VP microdomains in PS-b-P4VP thin films.

In Situ GISAXS Study on Solvent Vapour Induced Orientation Switching in PS-b-P4VP Block Copolymer Thin Films

Energy Technology Data Exchange (ETDEWEB)

Gowd, E Bhoje; Boehme, Marcus; Stamm, Manfred, E-mail: gowd@ipfdd.de, E-mail: bhojegowd@yahoo.com [Department of Nanostructured Materials Leibniz Institute of Polymer Research Dresden Hohe Strasse 6, 01069, Dresden (Germany)

2010-11-15

We investigated the orientation changes of cylindrical P4VP microdomains in PS-b-P4VP thin films upon annealing in different solvent vapours using the time-resolved in situ grazing-incidence small-angle X-ray scattering (GISAXS) for the first time. Swelling of perpendicular cylinders (C perpendicular) in a non-selective solvent vapours (chloroform) leads to the orientation change to in-plane cylinders (C//) and it occurs through a disordered state. On the other hand, swelling of perpendicular cylinders (C perpendicular) in a selective solvent vapours (1,4-dioxane) leads the morphological change from cylindrical to BCC spherical morphology. Solvent evaporation results in shrinkage of the matrix in the vertical direction and subsequently merges the spheres into the perpendicularly aligned cylinders. The selectivity of the solvent to constituting blocks and the solvent evaporation rate may be mainly responsible for such orientation change of cylindrical P4VP microdomains in PS-b-P4VP thin films.

The Ebola Virus Nucleoprotein Recruits the Host PP2A-B56 Phosphatase to Activate Transcriptional Support Activity of VP30

DEFF Research Database (Denmark)

Kruse, Thomas; Biedenkopf, Nadine; Hertz, Emil Peter Thrane

2018-01-01

Transcription of the Ebola virus genome depends on the viral transcription factor VP30 in its unphosphorylated form, but the underlying molecular mechanism of VP30 dephosphorylation is unknown. Here we show that the Ebola virus nucleoprotein (NP) recruits the host PP2A-B56 protein phosphatase......A-B56 and show that it suppresses Ebola virus transcription and infection. This work dissects the molecular mechanism of VP30 dephosphorylation by PP2A-B56, and it pinpoints this phosphatase as a potential target for therapeutic intervention....

Cardiotocography as a predictor of fetal outcome in women presenting with reduced fetal movement.

LENUS (Irish Health Repository)

Daly, Niamh

2011-09-05

OBJECTIVE: To examine the obstetric and perinatal outcomes of women presenting with reduced fetal movement (RFM) during the third trimester, specifically in relation to the diagnostic capacity of non-stress cardiotocography (CTG) used as the primary investigation in this clinical scenario. STUDY DESIGN: This was a retrospective population-based cohort study of pregnancy outcomes of all women ≥28 weeks\\' gestation with singleton pregnancies presenting during one calendar year with maternal perception of RFM, all of whom underwent CTG at presentation. Main outcome measures included: obstetric intervention (induction of labour, spontaneous vaginal delivery, operative vaginal delivery, emergency caesarean section), and perinatal outcome (subsequent perinatal death, low Apgar scores (<7(5)), neonatal resuscitation and NICU admission). RESULTS: In all, 524 women presented with RFM and a live fetus, representing 7% of the antenatal obstetric population; 284 women (54%) were nulliparous. The reassuring CTG group comprised 482 (92%) women in whom initial CTG was reassuring and 15 (3%) where a repeat tracing within 1h was reassuring. The non-reassuring\\/abnormal CTG group (n=27, 5%) either underwent emergency delivery or comprehensive serial fetal assessment; this group had significantly higher rates of emergency caesarean delivery, neonatal resuscitation and NICU admission; the incidence of small-for-gestational-age infants did not differ significantly. No perinatal death occurred in either group following CTG. CONCLUSION: Normal non-stress CTG is a reliable screening indicator of fetal wellbeing in women presenting with perception of RFM in the third trimester; abnormal pregnancy outcomes were more common when initial CTG was abnormal or persistently non-reassuring.

Structure and dynamics of Ebola virus matrix protein VP40 by a coarse-grained Monte Carlo simulation

Science.gov (United States)

Pandey, Ras; Farmer, Barry

Ebola virus matrix protein VP40 (consisting of 326 residues) plays a critical role in viral assembly and its functions such as regulation of viral transcription, packaging, and budding of mature virions into the plasma membrane of infected cells. How does the protein VP40 go through structural evolution during the viral life cycle remains an open question? Using a coarse-grained Monte Carlo simulation we investigate the structural evolution of VP40 as a function of temperature with the input of a knowledge-based residue-residue interaction. A number local and global physical quantities (e.g. mobility profile, contact map, radius of gyration, structure factor) are analyzed with our large-scale simulations. Our preliminary data show that the structure of the protein evolves through different state with well-defined morphologies which can be identified and quantified via a detailed analysis of structure factor.

The Host E3-Ubiquitin Ligase TRIM6 Ubiquitinates the Ebola Virus VP35 Protein and Promotes Virus Replication.

Science.gov (United States)

Bharaj, Preeti; Atkins, Colm; Luthra, Priya; Giraldo, Maria Isabel; Dawes, Brian E; Miorin, Lisa; Johnson, Jeffrey R; Krogan, Nevan J; Basler, Christopher F; Freiberg, Alexander N; Rajsbaum, Ricardo

2017-09-15

Ebola virus (EBOV), a member of the Filoviridae family, is a highly pathogenic virus that causes severe hemorrhagic fever in humans and is responsible for epidemics throughout sub-Saharan, central, and West Africa. The EBOV genome encodes VP35, an important viral protein involved in virus replication by acting as an essential cofactor of the viral polymerase as well as a potent antagonist of the host antiviral type I interferon (IFN-I) system. By using mass spectrometry analysis and coimmunoprecipitation assays, we show here that VP35 is ubiquitinated on lysine 309 (K309), a residue located on its IFN antagonist domain. We also found that VP35 interacts with TRIM6, a member of the E3-ubiquitin ligase tripartite motif (TRIM) family. We recently reported that TRIM6 promotes the synthesis of unanchored K48-linked polyubiquitin chains, which are not covalently attached to any protein, to induce efficient antiviral IFN-I-mediated responses. Consistent with this notion, VP35 also associated noncovalently with polyubiquitin chains and inhibited TRIM6-mediated IFN-I induction. Intriguingly, we also found that TRIM6 enhances EBOV polymerase activity in a minigenome assay and TRIM6 knockout cells have reduced replication of infectious EBOV, suggesting that VP35 hijacks TRIM6 to promote EBOV replication through ubiquitination. Our work provides evidence that TRIM6 is an important host cellular factor that promotes EBOV replication, and future studies will focus on whether TRIM6 could be targeted for therapeutic intervention against EBOV infection. IMPORTANCE EBOV belongs to a family of highly pathogenic viruses that cause severe hemorrhagic fever in humans and other mammals with high mortality rates (40 to 90%). Because of its high pathogenicity and lack of licensed antivirals and vaccines, EBOV is listed as a tier 1 select-agent risk group 4 pathogen. An important mechanism for the severity of EBOV infection is its suppression of innate immune responses. The EBOV VP35

Single Mutations in the VP2 300 Loop Region of the Three-Fold Spike of the Carnivore Parvovirus Capsid Can Determine Host Range

Science.gov (United States)

Organtini, Lindsey J.; Zhang, Sheng; Hafenstein, Susan L.; Holmes, Edward C.

2015-01-01

ABSTRACT Sylvatic carnivores, such as raccoons, have recently been recognized as important hosts in the evolution of canine parvovirus (CPV), a pandemic pathogen of domestic dogs. Although viruses from raccoons do not efficiently bind the dog transferrin receptor (TfR) or infect dog cells, a single mutation changing an aspartic acid to a glycine at capsid (VP2) position 300 in the prototype raccoon CPV allows dog cell infection. Because VP2 position 300 exhibits extensive amino acid variation among the carnivore parvoviruses, we further investigated its role in determining host range by analyzing its diversity and evolution in nature and by creating a comprehensive set of VP2 position 300 mutants in infectious clones. Notably, some position 300 residues rendered CPV noninfectious for dog, but not cat or fox, cells. Changes of adjacent residues (residues 299 and 301) were also observed often after cell culture passage in different hosts, and some of the mutations mimicked changes seen in viruses recovered from natural infections of alternative hosts, suggesting that compensatory mutations were selected to accommodate the new residue at position 300. Analysis of the TfRs of carnivore hosts used in the experimental evolution studies demonstrated that their glycosylation patterns varied, including a glycan present only on the domestic dog TfR that dictates susceptibility to parvoviruses. Overall, there were significant differences in the abilities of viruses with alternative position 300 residues to bind TfRs and infect different carnivore hosts, demonstrating that the process of infection is highly host dependent and that VP2 position 300 is a key determinant of host range. IMPORTANCE Although the emergence and pandemic spread of canine parvovirus (CPV) are well documented, the carnivore hosts and evolutionary pathways involved in its emergence remain enigmatic. We recently demonstrated that a region in the capsid structure of CPV, centered around VP2 position 300

Effect of reactive monomer on PS-b-P2VP film with UV irradiation

Science.gov (United States)

Kim, H. J.; Shin, D. M.

2012-03-01

Poly(styrene-b-2-vinyl pyridine) (PS-b-P2VP) lamellar film which is hydrophobic block hydrophilic polyelectrolyte block polymer of 52 kg/mol -b- 57 kg/mol and PS-b-P2VP film with reactive monomer (RM257) were prepared for photonic gel films. The lamellar stacks, which is alternating layer of hydrophilic and hydrophobic part of PS-b-P2VP. We reported about the influence of reactive monomer on those photonic gel films. Added reactive monomer photonic gel film had higher absorbance than pure photonic gel films. And band gaps of the lamellar films shifted by the time of UV light irradiation. That Photonic gel films were measured with the UV spectrophotometer. As a result the photonic gel film with reactive monomer had more clear color. The lamellar films were swollen by DI water, Ethyl alcohol (aq) and calcium carbonate solution. Since the domain spacing of dried photonic gel films were not showing any color in visible wavelength. The band gap of the lamellar films were drastically shifted to longer wavelength swollen by calcium carbonate solution (absorbance peak 565nm-->617nm). And the lamellar films were shifted to shorter wave length swollen by ethanol (absorbance peak 565nm-->497nm). So each Photonic gel film showed different color.

An Overview Of Weaving- Bodo Women In The Present Context.

Directory of Open Access Journals (Sweden)

Bibungsar Swargiary

2017-11-01

Full Text Available The Bodos are the early settler of the North-east India basically of Assam and the Bodos are also inhabitants of neighboring country outside of India like Nepal Bhutan and Bangladesh etc. The Bodos are most rich in their cultural activities. The weaving is the integral part of Bodos or women folk of the Bodo society. The women folk of the Bodo society are very expert in their weaving and the weaving is the small scale industry of the Bodo society. They weave their essential clothes in the loom with beautiful designing. In the present perspective they also earn money with selling their weaving clothes and make strong their family economy. The beautiful design clothes of women folk of the Bodo society not only popular among them but also getting memorable in other community also. Today their weaving activity comes to in commercial purpose and every woman have known to weave clothes very beautifully.

The VP3 factor from viruses of Birnaviridae family suppresses RNA silencing by binding both long and small RNA duplexes.

Directory of Open Access Journals (Sweden)

Adrian Valli

Full Text Available RNA silencing is directly involved in antiviral defense in a wide variety of eukaryotic organisms, including plants, fungi, invertebrates, and presumably vertebrate animals. The study of RNA silencing-mediated antiviral defences in vertebrates is hampered by the overlap with other antiviral mechanisms; thus, heterologous systems are often used to study the interplay between RNA silencing and vertebrate-infecting viruses. In this report we show that the VP3 protein of the avian birnavirus Infectious bursal disease virus (IBDV displays, in addition to its capacity to bind long double-stranded RNA, the ability to interact with double-stranded small RNA molecules. We also demonstrate that IBDV VP3 prevents the silencing mediated degradation of a reporter mRNA, and that this silencing suppression activity depends on its RNA binding ability. Furthermore, we find that the anti-silencing activity of IBDV VP3 is shared with the homologous proteins expressed by both insect- and fish-infecting birnaviruses. Finally, we show that IBDV VP3 can functionally replace the well-characterized HCPro silencing suppressor of Plum pox virus, a potyvirus that is unable to infect plants in the absence of an active silencing suppressor. Altogether, our results support the idea that VP3 protects the viral genome from host sentinels, including those of the RNA silencing machinery.

Expression of chicken parvovirus VP2 in chicken embryo fibroblasts requires codon optimization for production of naked DNA and vectored meleagrid herpesvirus type 1 vaccines.

Science.gov (United States)

Spatz, Stephen J; Volkening, Jeremy D; Mullis, Robert; Li, Fenglan; Mercado, John; Zsak, Laszlo

2013-10-01

Meleagrid herpesvirus type 1 (MeHV-1) is an ideal vector for the expression of antigens from pathogenic avian organisms in order to generate vaccines. Chicken parvovirus (ChPV) is a widespread infectious virus that causes serious disease in chickens. It is one of the etiological agents largely suspected in causing Runting Stunting Syndrome (RSS) in chickens. Initial attempts to express the wild-type gene encoding the capsid protein VP2 of ChPV by insertion into the thymidine kinase gene of MeHV-1 were unsuccessful. However, transient expression of a codon-optimized synthetic VP2 gene cloned into the bicistronic vector pIRES2-Ds-Red2, could be demonstrated by immunocytochemical staining of transfected chicken embryo fibroblasts (CEFs). Red fluorescence could also be detected in these transfected cells since the red fluorescent protein gene is downstream from the internal ribosome entry site (IRES). Strikingly, fluorescence could not be demonstrated in cells transiently transfected with the bicistronic vector containing the wild-type or non-codon-optimized VP2 gene. Immunocytochemical staining of these cells also failed to demonstrate expression of wild-type VP2, indicating that the lack of expression was at the RNA level and the VP2 protein was not toxic to CEFs. Chickens vaccinated with a DNA vaccine consisting of the bicistronic vector containing the codon-optimized VP2 elicited a humoral immune response as measured by a VP2-specific ELISA. This VP2 codon-optimized bicistronic cassette was rescued into the MeHV-1 genome generating a vectored vaccine against ChPV disease.

Crystal Structure of Marburg Virus VP40 Reveals a Broad, Basic Patch for Matrix Assembly and a Requirement of the N-Terminal Domain for Immunosuppression.

Science.gov (United States)

Oda, Shun-Ichiro; Noda, Takeshi; Wijesinghe, Kaveesha J; Halfmann, Peter; Bornholdt, Zachary A; Abelson, Dafna M; Armbrust, Tammy; Stahelin, Robert V; Kawaoka, Yoshihiro; Saphire, Erica Ollmann

2016-02-15

Marburg virus (MARV), a member of the filovirus family, causes severe hemorrhagic fever with up to 90% lethality. MARV matrix protein VP40 is essential for assembly and release of newly copied viruses and also suppresses immune signaling in the infected cell. Here we report the crystal structure of MARV VP40. We found that MARV VP40 forms a dimer in solution, mediated by N-terminal domains, and that formation of this dimer is essential for budding of virus-like particles. We also found the N-terminal domain to be necessary and sufficient for immune antagonism. The C-terminal domains of MARV VP40 are dispensable for immunosuppression but are required for virus assembly. The C-terminal domains are only 16% identical to those of Ebola virus, differ in structure from those of Ebola virus, and form a distinct broad and flat cationic surface that likely interacts with the cell membrane during virus assembly. Marburg virus, a cousin of Ebola virus, causes severe hemorrhagic fever, with up to 90% lethality seen in recent outbreaks. Molecular structures and visual images of the proteins of Marburg virus are essential for the development of antiviral drugs. One key protein in the Marburg virus life cycle is VP40, which both assembles the virus and suppresses the immune system. Here we provide the molecular structure of Marburg virus VP40, illustrate differences from VP40 of Ebola virus, and reveal surfaces by which Marburg VP40 assembles progeny and suppresses immune function. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

THE FOCAL STRUCTURE IN MANDARIN VP-ELLIPSIS: A CROSS-LINGUISTIC PERSPECTIVE

Directory of Open Access Journals (Sweden)

Ting-Chi Wei

2009-06-01

Full Text Available This paper argues that Soh’s (2007 ΣP analysis only partially explains polarity operation in Mandarin VP-ellipsis. With new examples of the use of the particle que ‘however’, a polarity contrast of ye ‘also’, we propose that there are two focus projections in VP-ellipsis. One is the contrastive FocP headed by ye or que higher than TP and the other is the polarity PolP headed by an affirmative polarity focus shi ‘be’ or a covert negative polarity focus lower than TP. Foc interacts with Pol by a way of polarity concord, which is responsible for the polarity symmetry or asymmetry across two conjuncts. We suggest that the polarity concord is achieved via the Agree operation (Chomsky 2000, 2001 in line with Watanabe’s (2004 feature copying analysis of the negative concord. A cross-linguistic investigation of languages of various word orders, including English (SVO, Japanese (SOV, Atayal (VOS, and Bunun (VSO lends support to this focus account.

Identification of a novel NLS of herpes simplex virus type 1 (HSV-1) VP19C and its nuclear localization is required for efficient production of HSV-1.

Science.gov (United States)

Li, You; Zhao, Lei; Wang, Shuai; Xing, Junji; Zheng, Chunfu

2012-09-01

Herpes simplex virus type 1 (HSV-1) triplex is a complex of three protein subunits, consisting of two copies of VP23 and one copy of VP19C. Here, we identified a non-classical NLS of VP19C between aa 50 and 61, and the nuclear import of VP19C was mediated by RanGTP and importin β1-, but not importin α5-, dependent pathway. Additionally, recombinant virus harbouring this NLS mutation (NLSm) replicates less efficiently as wild-type. These data strongly suggested that the nuclear import of VP19C is required for efficient HSV-1 production.

Re-Presenting Women and Leadership: A Methodological Journey

Science.gov (United States)

Wilkinson, Jane; Blackmore, Jill

2008-01-01

Research on women's leadership has tended to focus upon detailed micro studies of individual women's identity formation or, alternatively, to conduct macro studies of its broader discursive constructions within society. Both approaches, although providing helpful understandings of the issues surrounding constructions of women's leadership, are…

Immunogenicity of a DNA-launched replicon-based canine parvovirus DNA vaccine expressing VP2 antigen in dogs.

Science.gov (United States)

Dahiya, Shyam S; Saini, Mohini; Kumar, Pankaj; Gupta, Praveen K

2012-10-01

A replicon-based DNA vaccine encoding VP2 gene of canine parvovirus (CPV) was developed by cloning CPV-VP2 gene into a replicon-based DNA vaccine vector (pAlpha). The characteristics of a replicon-based DNA vaccine like, self-amplification of transcripts and induction of apoptosis were analyzed in transfected mammalian cells. When the pAlpha-CPV-VP2 was injected intradermal as DNA-launched replicon-based DNA vaccine in dogs, it induced CPV-specific humoral and cell mediated immune responses. The virus neutralization antibody and lymphocyte proliferative responses were higher than conventional CPV DNA vaccine and commercial CPV vaccine. These results indicated that DNA-launched replicon-based CPV DNA vaccine was effective in inducing both CPV-specific humoral and cellular immune responses and can be considered as effective alternative to conventional CPV DNA vaccine and commercial CPV vaccine. Crown Copyright © 2012. Published by Elsevier India Pvt Ltd. All rights reserved.

Concentration of acrylamide in a polyacrylamide gel affects VP4 gene coding assignment of group A equine rotavirus strains with P[12] specificity

Science.gov (United States)

2010-01-01

Background It is universally acknowledged that genome segment 4 of group A rotavirus, the major etiologic agent of severe diarrhea in infants and neonatal farm animals, encodes outer capsid neutralization and protective antigen VP4. Results To determine which genome segment of three group A equine rotavirus strains (H-2, FI-14 and FI-23) with P[12] specificity encodes the VP4, we analyzed dsRNAs of strains H-2, FI-14 and FI-23 as well as their reassortants by polyacrylamide gel electrophoresis (PAGE) at varying concentrations of acrylamide. The relative position of the VP4 gene of the three equine P[12] strains varied (either genome segment 3 or 4) depending upon the concentration of acrylamide. The VP4 gene bearing P[3], P[4], P[6], P[7], P[8] or P[18] specificity did not exhibit this phenomenon when the PAGE running conditions were varied. Conclusions The concentration of acrylamide in a PAGE gel affected VP4 gene coding assignment of equine rotavirus strains bearing P[12] specificity. PMID:20573245

The SBP-Box Gene VpSBP11 from Chinese Wild Vitis Is Involved in Floral Transition and Affects Leaf Development.

Science.gov (United States)

Hou, Hongmin; Yan, Xiaoxiao; Sha, Ting; Yan, Qin; Wang, Xiping

2017-07-13

Flowering occurs in angiosperms during a major developmental transition from vegetative growth to the reproductive phase. Squamosa promoter binding protein (SBP)-box genes have been found to play critical roles in regulating flower and fruit development, but their roles in grapevine have remained unclear. To better understand the functions of the grape SBP-box genes in both vegetative and reproductive growth phases, a full-length complementary DNA (cDNA) sequence of the putative SBP-box transcription factor gene, VpSBP11 , was obtained from Chinese wild grapevine Vitis pseudoreticulata Wen Tsai Wang (W. T. Wang) clone 'Baihe-35-1'. VpSBP11 encoded a putative polypeptide of 170 amino acids with a highly conserved SBP-domain with two zinc-binding sites of the Cx2C-x3-H-x11-C-x6-H (C2HCH) type and a nuclear localization signal. We confirmed that the VpSBP11 protein was targeted to the nucleus and possessed transcriptional activation activity by subcellular localization and trans -activation assay. Over-expression of VpSBP11 in Arabidopsis thaliana was shown to activate the FUL gene, and subsequently the AP1 and LFY genes, all of which were floral meristem identity genes, and to cause earlier flowering than in wild type (WT) plants. The pattern of vegetative growth was also different between the transgenic and WT plants. For example, in the VpSBP11 over-expressing transgenic plants, the number of rosette leaves was less than that of WT; the petiole was significantly elongated; and the rosette and cauline leaves curled upwards or downwards. These results were consistent with VpSBP11 acting as a transcription factor during the transition from the vegetative stage to the reproductive stage.

Objective and subjective image quality of primary and recurrent squamous cell carcinoma on head and neck low-tube-voltage 80-kVp computed tomography

Energy Technology Data Exchange (ETDEWEB)

Scholtz, Jan-Erik; Kaup, Moritz; Kraft, Johannes; Noeske, Eva-Maria; Schulz, Boris; Burck, Iris; Kerl, J.M.; Bauer, Ralf W.; Lehnert, Thomas; Vogl, Thomas J.; Wichmann, Julian L. [University Hospital Frankfurt, Department of Diagnostic and Interventional Radiology, Frankfurt (Germany); Scheerer, Friedrich [University Hospital Frankfurt, Department of Cranio-Maxillofacial and Plastic Facial Surgery, Frankfurt (Germany); Wagenblast, Jens [University Hospital Frankfurt, Department of Otolaryngology, Head and Neck Surgery, Frankfurt (Germany)

2015-03-26

To investigate low-tube-voltage 80-kVp computed tomography (CT) of head and neck primary and recurrent squamous cell carcinoma (SCC) regarding objective and subjective image quality. We retrospectively evaluated 65 patients (47 male, 18 female; mean age: 62.1 years) who underwent head and neck dual-energy CT (DECT) due to biopsy-proven primary (n = 50) or recurrent (n = 15) SCC. Eighty peak kilovoltage and standard blended 120-kVp images were compared. Attenuation and noise of malignancy and various soft tissue structures were measured. Tumor signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) were calculated. Subjective image quality was rated by three reviewers using 5-point grading scales regarding overall image quality, lesion delineation, image sharpness, and image noise. Radiation dose was assessed as CT dose index volume (CTDI{sub vol}). Interobserver agreement was calculated using intraclass correlation coefficient (ICC). Mean tumor attenuation (153.8 Hounsfield unit (HU) vs. 97.1 HU), SNR (10.7 vs. 8.3), CNR (8.1 vs. 4.8), and subjective tumor delineation (score, 4.46 vs. 4.13) were significantly increased (all P < 0.001) with 80-kVp acquisition compared to standard blended 120-kVp images. Noise of all measured structures was increased in 80-kVp acquisition (P < 0.001). Overall interobserver agreement was good (ICC, 0.86; 95 % confidence intervals: 0.82-0.89). CTDI{sub vol} was reduced by 48.7 % with 80-kVp acquisition compared to standard DECT (4.85 ± 0.51 vs. 9.94 ± 0.81 mGy cm, P < 0.001). Head and neck CT with low-tube-voltage 80-kVp acquisition provides increased tumor delineation, SNR, and CNR for CT imaging of primary and recurrent SCC compared to standard 120-kVp acquisition with an accompanying significant reduction of radiation exposure. (orig.)

Generation of recombinant monoclonal antibodies to study structure-function of envelope protein VP28 of white spot syndrome virus from shrimp

International Nuclear Information System (INIS)

Wang Yuzhen; Zhang Xiaohua; Yuan Li; Xu Tao; Rao Yu; Li Jia; Dai Heping

2008-01-01

White spot syndrome virus (WSSV) is a major pathogen in shrimp aquaculture. VP28 is one of the most important envelope proteins of WSSV. In this study, a recombinant antibody library, as single-chain fragment variable (scFv) format, displayed on phage was constructed using mRNA from spleen cells of mice immunized with full-length VP28 expressed in Escherichia coli. After several rounds of panning, six scFv antibodies specifically binding to the epitopes in the N-terminal, middle, and C-terminal regions of VP28, respectively, were isolated from the library. Using these scFv antibodies as tools, the epitopes in VP28 were located on the envelope of the virion by immuno-electron microscopy. Neutralization assay with these antibodies in vitro suggested that these epitopes may not be the attachment site of WSSV to host cell receptor. This study provides a new way to investigate the structure and function of the envelope proteins of WSSV

Ionic Salt Effect on the Phase Transition of PS-b-P2VP Copolymers

Science.gov (United States)

Kim, Bokyung; An, Hyungju; Ryu, Du Yeol; Kim, Jehan

2009-03-01

Solid-state electrolytes have long been considered as suitable candidates owing to the simple and easy processes for rechargeable battery manufactures, compared to conventional liquid electrolyte counterparts. Especially, polymer/salt systems involving PMMA and PVP complex forms have been studied since they provide stable electrochemical characteristics as well as mechanical properties. We studied the phase behavior of PS-b-P2VP upon the salt addition by small angle x-ray scattering (SAXS) and depolarized light scattering. Transition temperatures of block copolymer were significantly influenced by the salt addition in addition to the changes of d-spacings, which is caused by the effective coordinative interaction between P2VP block and salt. This study suggests a simple approach to solid-state block copolymer electrolytes.

Thin Crust and High Crustal Vp/Vs beneath the Central Armenia Plateau of the Lesser Caucasus

Science.gov (United States)

Tseng, T. L.; Lin, C. M.; Huang, B. S.; Karakhanyan, A.

2017-12-01

Armenia volcanic highland is part of the Lesser Caucasus directly connected with the East Anatolian Plateau to the west and Iranian Plateau to the east. Abundant Quaternary volcanoes in Armenia are the youngest among those associated with post-collision of Arabia-Eurasian since Miocene ( 11 Ma). In this study, teleseismic receiver functions were analyzed from a temporary array to constrain the crustal structures under Armenia and the vicinity. The results show that the Moho depth is shallowest beneath central Armenia where the estimated crustal thickness is 32 km with high averaged crustal Vp/Vs of 1.8-2.0 using H-κ technique. The high crustal Vp/Vs is distributed in a wider area but thin crust is confined more locally around stratovolcano Aragats, whose last eruption was about 0.5 Ma. High crustal Vp/Vs value approaching to 2.1 is found near East of volcano Ghegam complex and NW of volcano Ararat with last dated ages of 0.5 and <0.1 Ma, respectively. Such high Vp/Vs (2.0) cannot be explained without high mafic content and the presence of partial melt in the crust. The 1-D velocity models inverted demonstrate that the partial melt is more likely in the low-velocity layer of the lower crust. To support the unusually thin crust in central Armenia, it requires additional thermal buoyancy in the uppermost mantle which is consistent with regionally low Pn velocity found in previous studies. We propose that the volcanism here is facilitated by the stretches of lithosphere.

Expression of Recombinant Pichia pastoris VP1-2A and Multi-epitopes of Type O Foot-and-Mouth Disease Virus%O型口蹄疫病毒VP1-2A基因及多表位片段在毕赤酵母中的表达

Institute of Scientific and Technical Information of China (English)

袭莹; 朱战波; 金宁一; 胡博; 任静强; 刘存霞; 张军; 王鹤; 鲁会军

2010-01-01

利用毕赤酵母表达系统串联表达O型口蹄疫病毒(FMDV)VP1-2A基因及O型FMDV多表位片段(CTE),将O型FMDV vp1基因和CTE多表位片段克隆到毕赤酵母分泌性表达载体pPIC9K中,构建重组表达载体pPIC9K-VP1-2A-CTE并测序.经Sal I线性化后,通过电击转化法将重组质粒导入毕赤酵母GS115中,并对表达产物用SDS-PAGE和Western blot进行分析.重组质粒通过电转进入毕赤酵母后,能表达相对分子量为41.8KD(CTE)和26.5KD(VP1-2A)的蛋白,经Western blot分析,两种蛋白均具有抗原性.在毕赤酵母中成功地表达O型FMDV VP1-2A蛋白和多表位片段(CTE),为研制新型口蹄疫的基因工程疫苗奠定了基础.

Processing of the VP1/2A Junction Is Not Necessary for Production of Foot-and-Mouth Disease Virus Empty Capsids and Infectious Viruses: Characterization of “Self-Tagged” Particles

DEFF Research Database (Denmark)

Gullberg, Maria; Polacek, Charlotta; Bøtner, Anette

2013-01-01

The foot-and-mouth disease virus (FMDV) capsid protein precursor, P1-2A, is cleaved by 3Cpro to generate VP0, VP3, VP1, and the peptide 2A. The capsid proteins self-assemble into empty capsid particles or viruses which do not contain 2A. In a cell culture-adapted strain of FMDV (O1 Manisa [Lindholm...... the unmodified empty capsids in antigen enzyme-linked immunosorbent assays and integrin receptor binding assays. Furthermore, mutant viruses with uncleaved VP1-2A could be rescued in cells from full-length FMDV RNA transcripts encoding the K210E substitution in VP1. Thus, cleavage of the VP1/2A junction...

Cloning and expression of a sorghum gene with homology to maize vp1. Its potential involvement in pre-harvest sprouting resistance.

Science.gov (United States)

Carrari, F; Perez-Flore, L; Lijavetzky, D; Enciso, S; Sanchez, R; Benech-Arnold, R; Iusem, N

2001-04-01

Pre-harvest sprouting (PHS) in sorghum is related to the lack of a normal dormancy level during seed development and maturation. Based on previous evidence that seed dormancy in maize is controlled by the vp1 gene, we used a PCR-based approach to isolate two Sorghum bicolor genomic and cDNA clones from two genotypes exhibiting different PHS behaviour and sensitivity to abscisic acid (ABA). The two 699 amino acid predicted protein sequences differ in two residues at positions 341 (Gly or Cys within the repression domain) and 448 (Pro or Ser) and show over 80, 70 and 60% homology to maize, rice and oat VP1 proteins respectively. Expression analysis of the sorghum vp1 gene in the two lines shows a slightly higher level of vp1 mRNA in the embryos susceptible to PHS than in those resistant to PHS during embryogenesis. However, timing of expression was different between these genotypes during this developmental process. Whereas for the former the main peak of expression was observed at 20 days after pollination (DAP), the peak in the latter was found at later developmental stages when seed maturation was almost complete. Under favourable germination conditions and in the presence of fluridone (an inhibitor of ABA biosynthesis), sorghum vp1 mRNA showed to be consistently correlated with sensitivity to ABA but not with ABA content and dormancy.

Transmission of broad W/Rh and W/Al (target/filter) x-ray beams operated at 25-49 kVp through common shielding materials.

Science.gov (United States)

Li, Xinhua; Zhang, Da; Liu, Bob

2012-07-01

To provide transmission data for broad 25-39 kVp (kilovolt peak) W/Rh and 25-49 kVp W/Al (target/filter, W-tungsten, Rh-rhodium, and Al-aluminum) x-ray beams through common shielding materials, such as lead, concrete, gypsum wallboard, wood, steel, and plate glass. The unfiltered W-target x-ray spectra measured on a Selenia Dimensions system (Hologic Inc., Bedford, MA) set at 20-49 kVp were, respectively, filtered using 50-μm Rh and 700-μm Al, and were subsequently used for Monte Carlo calculations. The transmission of broad x-ray beams through shielding materials was simulated using Geant4 low energy electromagnetic physics package with photon- and electron-processes above 250 eV, including photoelectric effect, Compton scattering, and Rayleigh scattering. The calculated transmission data were fitted using Archer equation with a robust fitting algorithm. The transmission of broad x-ray beams through the above-mentioned shielding materials was calculated down to about 10(-5) for 25-39 kVp W/Rh and 25-49 kVp W/Al. The fitted results of α, β, and γ in Archer equation were provided. The α values of kVp ≥ 40 were approximately consistent with those of NCRP Report No. 147. These data provide inputs for the shielding designs of x-ray imaging facilities with W-anode x-ray beams, such as from Selenia Dimensions.

Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of the VP8* carbohydrate-binding protein of the human rotavirus strain Wa

International Nuclear Information System (INIS)

Kraschnefski, Mark J.; Scott, Stacy A.; Holloway, Gavan; Coulson, Barbara S.; Itzstein, Mark von; Blanchard, Helen

2005-01-01

The carbohydrate-binding component (VP8* 64–223 ) of the human Wa rotavirus spike protein has been overexpressed in E. coli, purified and crystallized in two different crystal forms. X-ray diffraction data have been collected that have enabled determination of the Wa VP8* 64–223 structure by molecular replacement. Rotaviruses exhibit host-specificity and the first crystallographic information on a rotavirus strain that infects humans is reported here. Recognition and attachment to host cells, leading to invasion and infection, is critically linked to the function of the outer capsid spike protein of the rotavirus particle. In some strains the VP8* component of the spike protein is implicated in recognition and binding of sialic-acid-containing cell-surface carbohydrates, thereby enabling infection by the virus. The cloning, expression, purification, crystallization and initial X-ray diffraction analysis of the VP8* core from human Wa rotavirus is reported. Two crystal forms (trigonal P3 2 21 and monoclinic P2 1 ) have been obtained and X-ray diffraction data have been collected, enabling determination of the VP8* 64–223 structure by molecular replacement

Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of the VP8* carbohydrate-binding protein of the human rotavirus strain Wa

Energy Technology Data Exchange (ETDEWEB)

Kraschnefski, Mark J.; Scott, Stacy A. [Institute for Glycomics, Griffith University (Gold Coast Campus), PMB 50 Gold Coast Mail Centre, Queensland 9726 (Australia); Holloway, Gavan; Coulson, Barbara S.; Itzstein, Mark von [Department of Microbiology and Immunology, The University of Melbourne, Victoria 3010 (Australia); Blanchard, Helen, E-mail: h.blanchard@griffith.edu.au [Institute for Glycomics, Griffith University (Gold Coast Campus), PMB 50 Gold Coast Mail Centre, Queensland 9726 (Australia)

2005-11-01

The carbohydrate-binding component (VP8*{sub 64–223}) of the human Wa rotavirus spike protein has been overexpressed in E. coli, purified and crystallized in two different crystal forms. X-ray diffraction data have been collected that have enabled determination of the Wa VP8*{sub 64–223} structure by molecular replacement. Rotaviruses exhibit host-specificity and the first crystallographic information on a rotavirus strain that infects humans is reported here. Recognition and attachment to host cells, leading to invasion and infection, is critically linked to the function of the outer capsid spike protein of the rotavirus particle. In some strains the VP8* component of the spike protein is implicated in recognition and binding of sialic-acid-containing cell-surface carbohydrates, thereby enabling infection by the virus. The cloning, expression, purification, crystallization and initial X-ray diffraction analysis of the VP8* core from human Wa rotavirus is reported. Two crystal forms (trigonal P3{sub 2}21 and monoclinic P2{sub 1}) have been obtained and X-ray diffraction data have been collected, enabling determination of the VP8*{sub 64–223} structure by molecular replacement.

Point mutation in calcium-binding domain of mouse polyomavirus VP1 protein does not prevent virus-like particle formation, but changes VP1 interactions with Saccharomyces cerevisiae cell structures

Czech Academy of Sciences Publication Activity Database

Adamec, T.; Palková, Zdena; Velková, K.; Štokrová, Jitka; Forstová, J.

2005-01-01

Roč. 5, 4-5 (2005), s. 331-340 ISSN 1567-1356 R&D Projects: GA ČR GA204/03/0593 Institutional research plan: CEZ:AV0Z5052915 Keywords : polyomavirus VP1 * Saccharomyces cerevisiae * heterologous expression Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.477, year: 2005

Generation and evaluation of a recombinant genotype VII Newcastle disease virus expressing VP3 protein of Goose parvovirus as a bivalent vaccine in goslings.

Science.gov (United States)

Wang, Jianzhong; Cong, Yanlong; Yin, Renfu; Feng, Na; Yang, Songtao; Xia, Xianzhu; Xiao, Yueqiang; Wang, Wenxiu; Liu, Xiufan; Hu, Shunlin; Ding, Chan; Yu, Shengqing; Wang, Chunfeng; Ding, Zhuang

2015-05-04

Newcastle disease virus (NDV) and Goose parvovirus (GPV) are considered to be two of the most important and widespread viruses infecting geese. In this study, we generated a recombinant rmNA-VP3, expressing GPV VP3 using a modified goose-origin NDV NA-1 by changing the multi-basic cleavage site motif RRQKR↓F of the F protein to the dibasic motif GRQGR↓L as that of the avirulent strain LaSota as a vaccine vector. Expression of the VP3 protein in rmNA-VP3 infected cells was detected by immunofluorescence and Western blot assay. The genetic stability was examined by serially passaging 10 times in 10-day-old embryonated SPF chicken eggs. Goslings were inoculated with rmNA-VP3 showed no apparent signs of disease and developed a strong GPV and NDV neutralizing antibodies response. This is the first study demonstrating that recombinant NDV has the potential to serve as bivalent live vaccine against Goose parvovirus and Newcastle disease virus infection in birds. Copyright © 2015 Elsevier B.V. All rights reserved.

Identification of antigenic regions on VP2 of African horsesickness virus serotype 3 by using phage-displayed epitope libraries.

Science.gov (United States)

Bentley, L; Fehrsen, J; Jordaan, F; Huismans, H; du Plessis, D H

2000-04-01

VP2 is an outer capsid protein of African horsesickness virus (AHSV) and is recognized by serotype-discriminatory neutralizing antibodies. With the objective of locating its antigenic regions, a filamentous phage library was constructed that displayed peptides derived from the fragmentation of a cDNA copy of the gene encoding VP2. Peptides ranging in size from approximately 30 to 100 amino acids were fused with pIII, the attachment protein of the display vector, fUSE2. To ensure maximum diversity, the final library consisted of three sub-libraries. The first utilized enzymatically fragmented DNA encoding only the VP2 gene, the second included plasmid sequences, while the third included a PCR step designed to allow different peptide-encoding sequences to recombine before ligation into the vector. The resulting composite library was subjected to immunoaffinity selection with AHSV-specific polyclonal chicken IgY, polyclonal horse immunoglobulins and a monoclonal antibody (MAb) known to neutralize AHSV. Antigenic peptides were located by sequencing the DNA of phages bound by the antibodies. Most antigenic determinants capable of being mapped by this method were located in the N-terminal half of VP2. Important binding areas were mapped with high resolution by identifying the minimum overlapping areas of the selected peptides. The MAb was also used to screen a random 17-mer epitope library. Sequences that may be part of a discontinuous neutralization epitope were identified. The amino acid sequences of the antigenic regions on VP2 of serotype 3 were compared with corresponding regions on three other serotypes, revealing regions with the potential to discriminate AHSV serotypes serologically.

Essential role of the unordered VP2 n-terminal domain of the parvovirus MVM capsid in nuclear assembly and endosomal enlargement of the virion fivefold channel for cell entry

Energy Technology Data Exchange (ETDEWEB)

Sanchez-Martinez, Cristina; Grueso, Esther [Centro de Biologia Molecular Severo Ochoa (CSIC-UAM), Universidad Autonoma de Madrid, 28049 Cantoblanco, Madrid (Spain); Carroll, Miles [Health Protection Agency, Centre for Emergency Preparedness and Response, Porton Down, Salisbury SP4 OJG, Wilts (United Kingdom); Rommelaere, Jean [Deutsches Krebsforschungszentrum Division F010, Im Neuenheimer Feld 242, D-69120 Heidelberg (Germany); Almendral, Jose M., E-mail: jmalmendral@cbm.uam.es [Centro de Biologia Molecular Severo Ochoa (CSIC-UAM), Universidad Autonoma de Madrid, 28049 Cantoblanco, Madrid (Spain)

2012-10-10

The unordered N-termini of parvovirus capsid proteins (Nt) are translocated through a channel at the icosahedral five-fold axis to serve for virus traffick. Heterologous peptides were genetically inserted at the Nt of MVM to study their functional tolerance to manipulations. Insertion of a 5T4-single-chain antibody at VP2-Nt (2Nt) yielded chimeric capsid subunits failing to enter the nucleus. The VEGFR2-binding peptide (V1) inserted at both 2Nt and VP1-Nt efficiently assembled in virions, but V1 disrupted VP1 and VP2 entry functions. The VP2 defect correlated with restricted externalization of V1-2Nt out of the coat. The specific infectivity of MVM and wtVP-pseudotyped mosaic MVM-V1 virions, upon heating and/or partial 2Nt cleavage, demonstrated that some 2Nt domains become intracellularly translocated out of the virus shell and cleaved to initiate entry. The V1 insertion defines a VP2-driven endosomal enlargement of the channel as an essential structural rearrangement performed by the MVM virion to infect.

Essential role of the unordered VP2 n-terminal domain of the parvovirus MVM capsid in nuclear assembly and endosomal enlargement of the virion fivefold channel for cell entry

International Nuclear Information System (INIS)

Sánchez-Martínez, Cristina; Grueso, Esther; Carroll, Miles; Rommelaere, Jean; Almendral, José M.

2012-01-01

The unordered N-termini of parvovirus capsid proteins (Nt) are translocated through a channel at the icosahedral five-fold axis to serve for virus traffick. Heterologous peptides were genetically inserted at the Nt of MVM to study their functional tolerance to manipulations. Insertion of a 5T4-single-chain antibody at VP2-Nt (2Nt) yielded chimeric capsid subunits failing to enter the nucleus. The VEGFR2-binding peptide (V1) inserted at both 2Nt and VP1-Nt efficiently assembled in virions, but V1 disrupted VP1 and VP2 entry functions. The VP2 defect correlated with restricted externalization of V1-2Nt out of the coat. The specific infectivity of MVM and wtVP-pseudotyped mosaic MVM-V1 virions, upon heating and/or partial 2Nt cleavage, demonstrated that some 2Nt domains become intracellularly translocated out of the virus shell and cleaved to initiate entry. The V1 insertion defines a VP2-driven endosomal enlargement of the channel as an essential structural rearrangement performed by the MVM virion to infect.

MRI in patients with tuberculous spondylitis presenting as vertebra plana: A retrospective analysis and review of literature

International Nuclear Information System (INIS)

Sureka, J.; Samuel, S.; Keshava, S.N.; Venkatesh, K.; Sundararaj, G.D.

2013-01-01

Aim: To present the magnetic resonance imaging (MRI) findings of 10 patients with histopathologically proven tuberculous spondylitis (TS) presenting as vertebra plana (VP) on radiographs. Material and methods: Radiographs of 451 adult TS patients were reviewed. In this consecutive series, there were 11 patients who presented as VP. MRI of 10 of these patients was available for review. Results: VP-like collapse of a single vertebral body of the dorsal spine with preserved endplates and disc was seen in all cases. Epidural, pre- and para-vertebral soft tissue was found in all patients. Epidural soft tissue presenting on sagittal images as a convexity of the posterior longitudinal ligament was also found in all the signal of which was different from the involved vertebra on axial images. All patients showed posterior element involvement, which was characterized by preserved cortical outline without expansion. Conclusion: TS presenting with VP-like collapse of the bone is rare, accounting for 2.4% of the cases in the present series. MRI may show a collapsed vertebra with preserved endplates and disc. MRI findings that are suggestive of TS include: (1) signal intensity of the epidural soft-tissue mass on axial images, which is different from the vertebral body; (2) presence of a thin, T2-weighted hypointense capsule of the para-vertebral soft tissue; (3) posterior element involvement characterized by intact hypointense cortical outline without expansion; and (4) involvement of the costovertebral joint.

Single-energy pediatric chest computed tomography with spectral filtration at 100 kVp: effects on radiation parameters and image quality

Energy Technology Data Exchange (ETDEWEB)

Bodelle, Boris; Fischbach, Constanze; Booz, Christian; Yel, Ibrahim; Frellesen, Claudia; Kaup, Moritz; Beeres, Martin; Vogl, Thomas J.; Scholtz, Jan-Erik [Goethe University of Frankfurt, Department of Diagnostic and Interventional Radiology, Frankfurt (Germany)

2017-06-15

Most of the applied radiation dose at CT is in the lower photon energy range, which is of limited diagnostic importance. To investigate image quality and effects on radiation parameters of 100-kVp spectral filtration single-energy chest CT using a tin-filter at third-generation dual-source CT in comparison to standard 100-kVp chest CT. Thirty-three children referred for a non-contrast chest CT performed on a third-generation dual-source CT scanner were examined at 100 kVp with a dedicated tin filter with a tube current-time product resulting in standard protocol dose. We compared resulting images with images from children examined using standard single-source chest CT at 100 kVp. We assessed objective and subjective image quality and compared radiation dose parameters. Radiation dose was comparable for children 5 years old and younger, and it was moderately decreased for older children when using spectral filtration (P=0.006). Effective tube current increased significantly (P=0.0001) with spectral filtration, up to a factor of 10. Signal-to-noise ratio and image noise were similar for both examination techniques (P≥0.06). Subjective image quality showed no significant differences (P≥0.2). Using 100-kVp spectral filtration chest CT in children by means of a tube-based tin-filter on a third-generation dual-source CT scanner increases effective tube current up to a factor of 10 to provide similar image quality at equivalent dose compared to standard single-source CT without spectral filtration. (orig.)

Single-energy pediatric chest computed tomography with spectral filtration at 100 kVp: effects on radiation parameters and image quality

International Nuclear Information System (INIS)

Bodelle, Boris; Fischbach, Constanze; Booz, Christian; Yel, Ibrahim; Frellesen, Claudia; Kaup, Moritz; Beeres, Martin; Vogl, Thomas J.; Scholtz, Jan-Erik

2017-01-01

Most of the applied radiation dose at CT is in the lower photon energy range, which is of limited diagnostic importance. To investigate image quality and effects on radiation parameters of 100-kVp spectral filtration single-energy chest CT using a tin-filter at third-generation dual-source CT in comparison to standard 100-kVp chest CT. Thirty-three children referred for a non-contrast chest CT performed on a third-generation dual-source CT scanner were examined at 100 kVp with a dedicated tin filter with a tube current-time product resulting in standard protocol dose. We compared resulting images with images from children examined using standard single-source chest CT at 100 kVp. We assessed objective and subjective image quality and compared radiation dose parameters. Radiation dose was comparable for children 5 years old and younger, and it was moderately decreased for older children when using spectral filtration (P=0.006). Effective tube current increased significantly (P=0.0001) with spectral filtration, up to a factor of 10. Signal-to-noise ratio and image noise were similar for both examination techniques (P≥0.06). Subjective image quality showed no significant differences (P≥0.2). Using 100-kVp spectral filtration chest CT in children by means of a tube-based tin-filter on a third-generation dual-source CT scanner increases effective tube current up to a factor of 10 to provide similar image quality at equivalent dose compared to standard single-source CT without spectral filtration. (orig.)

The Effect of Reproductive Factors on Breast Cancer Presentation in Women Who Are BRCA Mutation Carrier.

Science.gov (United States)

Kim, Ju-Yeon; Moon, Hyeong-Gon; Kang, Young-Joon; Han, Wonshik; Noh, Woo-Chul; Jung, Yongsik; Moon, Byung-In; Kang, Eunyoung; Park, Sung-Shin; Lee, Min Hyuk; Park, Bo Young; Lee, Jong Won; Noh, Dong-Young

2017-09-01

Germline mutations in the BRCA1 and BRCA2 genes confer increased risks for breast cancers. However, the clinical presentation of breast cancer among women who are carriers of the BRCA1 or BRCA2 ( BRCA1/2 carriers) mutations is heterogenous. We aimed to identify the effects of the reproductive histories of women with the BRCA1/2 mutations on the clinical presentation of breast cancer. We retrospectively analyzed clinical data on women with proven BRCA1 and BRCA2 mutations who were recruited to the Korean Hereditary Breast Cancer study, from 2007 to 2014. Among the 736 women who were BRCA1/2 mutation carriers, a total of 483 women had breast cancers. Breast cancer diagnosis occurred at significantly younger ages in women who experienced menarche at ≤14 years of age, compared to those who experienced menarche at >14 years of age (37.38±7.60 and 43.30±10.11, respectively, p women with the BRCA2 mutation. The prevalence of advanced stages (stage II or III vs. stage I) of disease in parous women was higher than in nulliparous women (68.5% vs. 55.2%, p =0.043). This association was more pronounced in women with the BRCA2 mutation (hazard ratio, 2.67; p =0.014). Our results suggest that reproductive factors, such as the age of onset of menarche and the presence of parity, are associated with the clinical presentation patterns of breast cancer in BRCA1/2 mutation carriers.

Processing of the VP1/2A junction is not necessary for production of foot-and-mouth disease virus empty capsids and infectious viruses: characterization of "self-tagged" particles.

Science.gov (United States)

Gullberg, Maria; Polacek, Charlotta; Bøtner, Anette; Belsham, Graham J

2013-11-01

The foot-and-mouth disease virus (FMDV) capsid protein precursor, P1-2A, is cleaved by 3C(pro) to generate VP0, VP3, VP1, and the peptide 2A. The capsid proteins self-assemble into empty capsid particles or viruses which do not contain 2A. In a cell culture-adapted strain of FMDV (O1 Manisa [Lindholm]), three different amino acid substitutions (E83K, S134C, and K210E) were identified within the VP1 region of the P1-2A precursor compared to the field strain (wild type [wt]). Expression of the O1 Manisa P1-2A (wt or with the S134C substitution in VP1) plus 3C(pro), using a transient expression system, resulted in efficient capsid protein production and self-assembly of empty capsid particles. Removal of the 2A peptide from the capsid protein precursor had no effect on capsid protein processing or particle assembly. However, modification of E83K alone abrogated particle assembly with no apparent effect on protein processing. Interestingly, the K210E substitution, close to the VP1/2A junction, completely blocked processing by 3C(pro) at this cleavage site, but efficient assembly of "self-tagged" empty capsid particles, containing the uncleaved VP1-2A, was observed. These self-tagged particles behaved like the unmodified empty capsids in antigen enzyme-linked immunosorbent assays and integrin receptor binding assays. Furthermore, mutant viruses with uncleaved VP1-2A could be rescued in cells from full-length FMDV RNA transcripts encoding the K210E substitution in VP1. Thus, cleavage of the VP1/2A junction is not essential for virus viability. The production of such engineered self-tagged empty capsid particles may facilitate their purification for use as diagnostic reagents and vaccines.

Extreme Mutation Tolerance: Nearly Half of the Archaeal Fusellovirus Sulfolobus Spindle-Shaped Virus 1 Genes Are Not Required for Virus Function, Including the Minor Capsid Protein Gene vp3.

Science.gov (United States)

Iverson, Eric A; Goodman, David A; Gorchels, Madeline E; Stedman, Kenneth M

2017-05-15

Viruses infecting the Archaea harbor a tremendous amount of genetic diversity. This is especially true for the spindle-shaped viruses of the family Fuselloviridae , where >90% of the viral genes do not have detectable homologs in public databases. This significantly limits our ability to elucidate the role of viral proteins in the infection cycle. To address this, we have developed genetic techniques to study the well-characterized fusellovirus Sulfolobus spindle-shaped virus 1 (SSV1), which infects Sulfolobus solfataricus in volcanic hot springs at 80°C and pH 3. Here, we present a new comparative genome analysis and a thorough genetic analysis of SSV1 using both specific and random mutagenesis and thereby generate mutations in all open reading frames. We demonstrate that almost half of the SSV1 genes are not essential for infectivity, and the requirement for a particular gene correlates well with its degree of conservation within the Fuselloviridae The major capsid gene vp1 is essential for SSV1 infectivity. However, the universally conserved minor capsid gene vp3 could be deleted without a loss in infectivity and results in virions with abnormal morphology. IMPORTANCE Most of the putative genes in the spindle-shaped archaeal hyperthermophile fuselloviruses have no sequences that are clearly similar to characterized genes. In order to determine which of these SSV genes are important for function, we disrupted all of the putative genes in the prototypical fusellovirus, SSV1. Surprisingly, about half of the genes could be disrupted without destroying virus function. Even deletions of one of the known structural protein genes that is present in all known fuselloviruses, vp3 , allows the production of infectious viruses. However, viruses lacking vp3 have abnormal shapes, indicating that the vp3 gene is important for virus structure. Identification of essential genes will allow focused research on minimal SSV genomes and further understanding of the structure of

Differences in clinical presentation of ankylosing spondylitis in men and women

Directory of Open Access Journals (Sweden)

Hossain Soleymani Salehabadi

2016-10-01

Full Text Available Background: Ankylosing spondylitis (AS is an inflammatory disease that mainly affects axial skeleton of the body and ankylosing spondylitis ligaments around the spine at the junction of the spine are inflamed, because the disease is progressive and can lead to significantly cause of disability and the studies could provide a mechanism for the early detection of the disease or help determine when to start treatment, the difference in clinical presentations of AS in men and women is indicative of potential effect of gender on severity of the disease. This study was conducted with the aim to investigate the effect of gender on severity of AS. Methods: In a cross-sectional study, one hundred and fifteen patients with ankylosing spondylitis who referred to Yazd Rheumatology Clinic between 2001 and 2013 were evaluated. Sampling was performed using non-random convenient method. The most important variables studied included demographic data, clinical presentation, radiographic stage of sacroiliac involvement, and laboratory data extracted from patients’ files and recorded in questionnaires. Results: Both groups according to age at diagnosis, presence of enteritis, peripheral joint involvement and laboratory data such as C-reactive protein (CRP, erythrocyte sedimentation rate (ESR and hemoglobin were matched. Inflammatory neck pain was more prevalent in men than in women (77.2% against 51.8%; P< 0.05. Sacroiliac radiographic study revealed stage 1 involvement in 11.3% of men and 37% of women (P= 0.009, and stage 4 in 27.2% of men and 3.7% of women (P< 0.001, with a significant difference. Conclusion: According to the results of the study, the time between age of onset and age at diagnosis, inflammatory pain in the neck and advanced stage in men than in women was higher. Although these findings suggest that gender may have an impact on the pattern and severity of AS but the time delay in diagnosis as a disease affecting the intensity and pattern should

Value of 100 kVp scan with sinogram-affirmed iterative reconstruction algorithm on a single-source CT system during whole-body CT for radiation and contrast medium dose reduction: an intra-individual feasibility study.

Science.gov (United States)

Nagayama, Y; Nakaura, T; Oda, S; Tsuji, A; Urata, J; Furusawa, M; Tanoue, S; Utsunomiya, D; Yamashita, Y

2018-02-01

To perform an intra-individual investigation of the usefulness of a contrast medium (CM) and radiation dose-reduction protocol using single-source computed tomography (CT) combined with 100 kVp and sinogram-affirmed iterative reconstruction (SAFIRE) for whole-body CT (WBCT; chest-abdomen-pelvis CT) in oncology patients. Forty-three oncology patients who had undergone WBCT under both 120 and 100 kVp protocols at different time points (mean interscan intervals: 98 days) were included retrospectively. The CM doses for the 120 and 100 kVp protocols were 600 and 480 mg iodine/kg, respectively; 120 kVp images were reconstructed with filtered back-projection (FBP), whereas 100 kVp images were reconstructed with FBP (100 kVp-F) and the SAFIRE (100 kVp-S). The size-specific dose estimate (SSDE), iodine load and image quality of each protocol were compared. The SSDE and iodine load of 100 kVp protocol were 34% and 21%, respectively, lower than of 120 kVp protocol (SSDE: 10.6±1.1 versus 16.1±1.8 mGy; iodine load: 24.8±4versus 31.5±5.5 g iodine, p<0.01). Contrast enhancement, objective image noise, contrast-to-noise-ratio, and visual score of 100 kVp-S were similar to or better than of 120 kVp protocol. Compared with the 120 kVp protocol, the combined use of 100 kVp and SAFIRE in WBCT for oncology assessment with an SSCT facilitated substantial reduction in the CM and radiation dose while maintaining image quality. Copyright © 2017 The Royal College of Radiologists. Published by Elsevier Ltd. All rights reserved.

Determinants of the VP1/2A junction cleavage by the 3C protease in foot-and-mouth disease virus infected cells

DEFF Research Database (Denmark)

Kristensen, Thea; Normann, Preben; Gullberg, Maria

2017-01-01

. Interestingly, in contrast to the serotype O virus results, no second site mutations occurred within the VP1 coding region of serotype A viruses with the blocked VP1/2A cleavage site. However, some of these viruses acquired changes in the 2C protein that is involved in enterovirus morphogenesis. These results...

Analysis of the full-length VP2 protein of canine parvoviruses circulating in Hungary.

Science.gov (United States)

Cságola, Attila; Varga, Szilvia; Lőrincz, Márta; Tuboly, Tamás

2014-09-01

In recent years, the number of cases of disease caused by canine parvovirus 2 (CPV-2) in vaccinated dogs has increased. The aim of the present study was to identify CPV-2 strains present in Hungary. Forty-two out of 50 faecal specimens examined were positive, and 25 VP2 sequences were determined and analysed. Based on the current classification, the Hungarian viruses belong to New CPV-2a type, except two viruses that are recombinants of vaccine viruses and CPV-2a strains. The Tyr324Ile alteration was detected for the first time in Europe, and a "Hungarian-specific" substitution (Ala516Thr) was also identified in this study. The immunologically important parts of the currently spreading canine parvoviruses were examined and found to differ greatly from the vaccine strains that are widely used in Hungary.

Genital tuberculosis in postmenopausal women with variable clinical presentations: A report of 3 cases

Directory of Open Access Journals (Sweden)

Ashima Arora

2018-04-01

Full Text Available Genital tuberculosis is usually diagnosed in young women being assessed for infertility. After menopause it usually presents with symptoms resembling endometrial malignancy, such as postmenopausal bleeding, persistent vaginal discharge and pyometra. The diagnosis is made by detection of acid-fast bacilli on microscopy or bacteriological culture and/or presence of epithelioid granuloma on biopsy. Anti-tubercular therapy involves the use of rifampicin, isoniazid, pyrazinamide and ethambutol. Surgery is indicated if a pelvic mass and recurrence of pain or bleeding persist after 9 months of treatment. Three cases of genital tuberculosis in postmenopausal women with different clinical presentations are reported. The first woman presented with ascites and weight loss. The second had postmenopausal bleeding with a pipelle biopsy suggestive of endometrial intraepithelial neoplasia. The third presented with weight loss and a palpable abdominal mass. Pelvic malignancy was initially suspected but a diagnosis of tuberculosis was made following pre-operative endometrial biopsy, bacteriological culture and intra-operative frozen section. All three women responded to anti-tubercular therapy. Keywords: Genital tuberculosis, Postmenopausal, Endometrial intraepithelial neoplasia, Anti-tubercular therapy, Endometrial biopsy

Depression in African women presenting for psychological services ...

African Journals Online (AJOL)

one in five women.2 Depression in women, world wide, seems to manifest mostly as .... Results. Demographic distribution. A total of 54 clinical files were analyzed. The patients' .... business can help and will reduce the stigma associated with.

Assembly of Ebola virus matrix protein VP40 is regulated by latch-like properties of N and C terminal tails.

Directory of Open Access Journals (Sweden)

Leslie P Silva

Full Text Available The matrix protein VP40 coordinates numerous functions in the viral life cycle of the Ebola virus. These range from the regulation of viral transcription to morphogenesis, packaging and budding of mature virions. Similar to the matrix proteins of other nonsegmented, negative-strand RNA viruses, VP40 proceeds through intermediate states of assembly (e.g. octamers but it remains unclear how these intermediates are coordinated with the various stages of the life cycle. In this study, we investigate the molecular basis of synchronization as governed by VP40. Hydrogen/deuterium exchange mass spectrometry was used to follow induced structural and conformational changes in VP40. Together with computational modeling, we demonstrate that both extreme N and C terminal tail regions stabilize the monomeric state through a direct association. The tails appear to function as a latch, released upon a specific molecular trigger such as RNA ligation. We propose that triggered release of the tails permits the coordination of late-stage events in the viral life cycle, at the inner membrane of the host cell. Specifically, N-tail release exposes the L-domain motifs PTAP/PPEY to the transport and budding complexes, whereas triggered C-tail release could improve association with the site of budding.

Writing women into medical history in the 1930s: Kate Campbell Hurd-Mead and "medical women" of the past and present.

Science.gov (United States)

Appel, Toby A

2014-01-01

Kate Campbell Hurd-Mead (1867–1941), a leader among second-generation women physicians in America, became a pioneer historian of women in medicine in the 1930s. The coalescence of events in her personal life, the declining status of women in medicine, and the growing significance of the new and relatively open field of history of medicine all contributed to this transformation in her career. While she endeavored to become part of the community of male physicians who wrote medical history, her primary identity remained that of a “medical woman.” For Hurd-Mead, the history of women in the past not only filled a vital gap in scholarship but served practical ends that she had earlier pursued by other means—those of inspiring and advancing the careers of women physicians of the present day, promoting organizations of women physicians, and advocating for equality of opportunity in the medical profession.

Injected phage-displayed-VP28 vaccine reduces shrimp Litopenaeus vannamei mortality by white spot syndrome virus infection.

Science.gov (United States)

Solís-Lucero, G; Manoutcharian, K; Hernández-López, J; Ascencio, F

2016-08-01

White spot syndrome virus (WSSV) is the most important viral pathogen for the global shrimp industry causing mass mortalities with huge economic losses. Recombinant phages are capable of expressing foreign peptides on viral coat surface and act as antigenic peptide carriers bearing a phage-displayed vaccine. In this study, the full-length VP28 protein of WSSV, widely known as potential vaccine against infection in shrimp, was successfully cloned and expressed on M13 filamentous phage. The functionality and efficacy of this vaccine immunogen was demonstrated through immunoassay and in vivo challenge studies. In ELISA assay phage-displayed VP28 was bind to Litopenaeus vannamei immobilized hemocyte in contrast to wild-type M13 phage. Shrimps were injected with 2 × 10(10) cfu animal(-1) single dose of VP28-M13 and M13 once and 48 h later intramuscularly challenged with WSSV to test the efficacy of the vaccine against the infection. All dead challenged shrimps were PCR WSSV-positive. The accumulative mortality of the vaccinated and challenged shrimp groups was significantly lower (36.67%) than the unvaccinated group (66.67%). Individual phenoloxidase and superoxide dismutase activity was assayed on 8 and 48 h post-vaccination. No significant difference was found in those immunological parameters among groups at any sampled time evaluated. For the first time, phage display technology was used to express a recombinant vaccine for shrimp. The highest percentage of relative survival in vaccinated shrimp (RPS = 44.99%) suggest that the recombinant phage can be used successfully to display and deliver VP28 for farmed marine crustaceans. Copyright © 2016 Elsevier Ltd. All rights reserved.

Genetic characterization of type 2a canine parvoviruses from Taiwan reveals the emergence of an Ile324 mutation in VP2

Science.gov (United States)

2014-01-01

Background Canine parvovirus 2 (CPV 2) is a major infectious cause of mortality in puppies. The characteristic symptom of CPV 2 disease is intestinal hemorrhage with severe bloody diarrhea. Soon after CPV was first recognized in the late 1970s, the original virus, CPV 2, was replaced in the canine population by strains carrying minor antigenic variants (termed 2a, 2b, and 2c) of the VP2 gene that could be distinguished using monoclonal antibodies and molecular analyses. Here, we provide an updated molecular characterization of the CPV 2 circulating in Taiwan. Methods In this study, 28 isolates of CPV 2 from 144 dogs with suspected CPV infection were obtained from northern, central, and southern Taiwan from 2008 to 2012 and screened by PCR. The 28 isolates were sequenced, and a phylogenetic analysis of the VP2 gene was performed. Results Of the 28 Taiwanese CPV 2 isolates, 15 were identified as new CPV 2a, and 13 were identified as new CPV 2b. Compared to the reference CPV 2a, all 15 of the CPV 2a sequences collected in this study contain an Ile324 mutation caused by a TAT to ATT mutation at nucleotides 970–972 of the VP2 gene. Conclusion Our VP2 sequence data revealed that both types are currently prevalent CPV 2 field strains circulating in Taiwan, and a unique Ile324 VP2 mutation was found in our Taiwanese CPV 2a isolates and recent Asian isolates. CPV 2c was not observed in this study. PMID:24568207

Factors influencing the type of health problems presented by women in general practice: differences between women's health care and regular health care.

NARCIS (Netherlands)

Brink-Muinen, A. van den; Bensing, J.M.

1996-01-01

Objective: Differences between health problems presented by women (aged 20-45) to female "women's health care" doctors and both female and male regular health care doctors were investigated. This article explores the relationship of patients' roles (worker, partner, or parent) and the type of health

A descriptive study of women presenting to an obstetric triage unit with no prenatal care.

Science.gov (United States)

Knight, Erin; Morris, Margaret; Heaman, Maureen

2014-03-01

To describe women presenting to an obstetric triage unit with no prenatal care (PNC), to identify gaps in care, and to compare care provided to World Health Organization (WHO) standards. We reviewed the charts of women who gave birth at Women's Hospital in Winnipeg and were discharged between April 1, 2008, and March 31, 2011, and identified those whose charts were coded with ICD-10 code Z35.3 (inadequate PNC) or who had fewer than 2 PNC visits. Three hundred eighty-two charts were identified, and sociodemographic characteristics, PNC history, investigations, and pregnancy outcomes were recorded. The care provided was compared with WHO guidelines. One hundred nine women presented to the obstetric triage unit with no PNC; 96 (88.1%) were in the third trimester. Only 39 women (35.8%) received subsequent PNC, with care falling short of WHO standards. Gaps in PNC included missing time-sensitive screening tests, mid-stream urine culture, and Chlamydia and gonorrhea testing. The mean maternal age was 26.1 years, and 93 women (85.3%) were multigravidas. More than one half of the women (51.4%) were involved with Child and Family Services, 64.2% smoked, 33.0% drank alcohol, and 32.1% used illicit drugs during pregnancy. Two thirds of the women (66.2%) lived in inner-city Winnipeg. Only 63.0% of neonates showed growth appropriate for gestational age. Two pregnancies ended in stillbirth; there was one neonatal death, and over one third of the births were preterm. Most women who present with no PNC do so late in pregnancy, proceed to deliver with little or no additional PNC, and have high rates of adverse outcomes. Thus, efforts to improve PNC must focus on facilitating earlier entry into care. This would also improve compliance with WHO guidelines for continuing care. Treatment protocols could improve gaps in obtaining urine culture and in Chlamydia and gonorrhea testing.

猪轮状病毒重组VP8*蛋白的原核表达及免疫原性分析%Prokaryotic Expression and Immunogenic Analysis of △VP8* of Porcine Rotavirus Gottfried Strain

Institute of Scientific and Technical Information of China (English)

毕艳铎; 魏晓曼; 冉旭华; 曹思; 张峣; 张玲玲; 苗艳; 闻晓波

2015-01-01

为了分析原核表达的猪轮状病毒(PRV) Gottfried株截短的VP8*(△VP8* aa 64-223)重组蛋白的免疫原性,通过构建原核表达载体pET-28a-Gottfried-△VP8*(aa 64-223),转化大肠杆菌BL21 (DE3)感受态细胞,IPTG诱导表达、经SDS-PAGE和Western blotting检测,并纯化后,肌肉免疫Balb/c小鼠,证明重组蛋白相对分子质量约25.0 kDa,以包涵体和可溶性两种形式存在,纯化的重组蛋白纯度在95％以上,可溶性蛋白产量可达14～15 mg·L-1.ELISA方法检测表明该重组蛋白可以诱导高滴度的特异性抗体,证明其具有良好的免疫原性,为进一步研制PRV亚单位疫苗及建立ELISA诊断方法奠定基础.

Inclusion of a universal tetanus toxoid CD4(+) T cell epitope P2 significantly enhanced the immunogenicity of recombinant rotavirus ΔVP8* subunit parenteral vaccines.

Science.gov (United States)

Wen, Xiaobo; Wen, Ke; Cao, Dianjun; Li, Guohua; Jones, Ronald W; Li, Jianping; Szu, Shousun; Hoshino, Yasutaka; Yuan, Lijuan

2014-07-31

Currently available live oral rotavirus vaccines, Rotarix(®) and RotaTeq(®), are highly efficacious in developed countries. However, the immunogenicity and efficacy of such vaccines in some developing countries are low. We reported previously that bacterially-expressed rotavirus ΔVP8* subunit vaccine candidates with P[8], P[4] or P[6] specificity elicited high-titer virus neutralizing antibodies in animals immunized intramuscularly. Of note was the finding that antibodies induced with the P[8]ΔVP8* vaccine neutralized both homotypic P[8] and heterotypic P[4] rotavirus strains to high titer. To further improve its vaccine potential, a tetanus toxoid universal CD4(+) T cell epitope P2 was introduced into P[8] or P[6]ΔVP8* construct. The resulting recombinant fusion proteins expressed in Escherichia coli were of high solubility and were produced with high yield. Two doses (10 or 20 μg/dose) of the P2-P[8]ΔVP8* vaccine or P2-P[6]ΔVP8* vaccine with aluminum phosphate adjuvant elicited significantly higher geometric mean homologous neutralizing antibody titers than the vaccines without P2 in intramuscularly immunized guinea pigs. Interestingly, high levels of neutralizing antibody responses induced in guinea pigs with 3 doses of the P2-P[8]ΔVP8* vaccine persisted for at least 6 months. Furthermore, in the gnotobiotic piglet challenge study, three intramuscular doses (50 μg/dose) of the P2-P[8]ΔVP8* vaccine with aluminum phosphate adjuvant significantly delayed the onset of diarrhea and significantly reduced the duration of diarrhea and the cumulative diarrhea score after oral challenge with virulent human rotavirus Wa (G1P[8]) strain. The P2-P[8]ΔVP8* vaccine induced serum virus neutralizing antibody and VP4-specific IgG antibody production prechallenge, and primed the pigs for higher antibody and intestinal and systemic virus-specific IFN-γ producing CD4(+) T cell responses postchallenge. These two subunit vaccines could be used at a minimum singly or

ρ-ω mixing in J/ψ→VP decays

International Nuclear Information System (INIS)

Wang, Dong; Ban, Yong; Li, Gang

2014-01-01

The study of ρ-ω mixing has mainly focused on vector meson decays with isospin I=1, namely the ρ(ω)→π + π − process. In this paper, we present a study of ρ-ω mixing in ρ(ω)→π + π − π 0 (I=0) using a flavor parameterization model for the J/ψ→VP process. By fitting a theoretical framework to PDG data, we obtain the SU(3)-breaking effect parameters s V =0.03±0.12, s P =0.17±0.17 and the ρ-ω mixing polarization operator Π ρω =(0.006±0.011)  GeV 2 . New values are found for the branching ratios when the mixing effect is incorporated: Br(J/ψ→ωπ 0 )=(3.64±0.37)×10 −4 , Br(J/ψ→ωη)=(1.48±0.17)×10 −3 , Br(J/ψ→ωη ′ )=(1.55±0.56)×10 −4 , these are different from the corresponding PDG2012 values by 19%, 15% and 15%, respectively

Depression in African women presenting for psychological services ...

African Journals Online (AJOL)

Depression in these women was related to poverty, overcrowding, unemployment, high levels of crime, lack of services and sexual abuse. Conclusion: African women return to a social context within which depression is increasingly prevalent. Establishing psychological services relevant to needs as well as means of ...

Isolation and sequence analysis of the complete NS1 and VP2 genes of canine parvovirus from domestic dogs in 2013 and 2014 in China.

Science.gov (United States)

Wang, Hualei; Jin, Hongli; Li, Qian; Zhao, Guoxing; Cheng, Nan; Feng, Na; Zheng, Xuexing; Wang, Jianzhong; Zhao, Yongkun; Li, Ling; Cao, Zengguo; Yan, Feihu; Wang, Lina; Wang, Tiecheng; Gao, Yuwei; Yang, Songtao; Xia, Xianzhu

2016-02-01

Canine parvovirus (CPV) can cause severe disease in animals and continuously generates new variant and recombinant strains in dogs that have a strong impact on sanitation. It is therefore necessary to investigate epidemic CPV strains to improve our understanding of CPV transmission and epidemic behavior. However, most studies have focused on the analysis of VP2, and therefore, information about recombination and relationships between strains is still lacking. Here, 14 strains of CPV were isolated from domestic dogs suspected of hosting CPV between 2013 and 2014 in China. The complete NS1 and VP2 genes were sequenced and analyzed. The results suggest that the new CPV-2a and new CPV-2b types are the prevalent strains in China. In addition to a few mutations (residues 19, 544, 545, 572 and 583 of NS1 and residues 267, 370, 377 and 440 of VP2) that were preserved during transmission, new mutations (residues 60, 630 of NS1, and residues 21, 310 of VP2) were found in the isolated strains. A phylogenetic tree based on VP2 sequences illustrated that the new CPV-2a and new CPV-2b strains from China form single clusters that are distinct from lineages from other countries. Moreover, recombination between the new CPV-2a and new CPV-2b types was also identified in the isolated strains. Due to differences in selection pressures or recombination, there were a small number of inconsistencies between the phylogenetic trees for VP2 and NS1, which indicated that phylogenetic relationships based on VP2 might not be representative of those based on NS1. The data indicated that mutations and recombination are constantly occurring along with the spread of CPV in China.

Interpretation of Pronouns in VP-Ellipsis Constructions in Dutch Broca's and Wernicke's Aphasia

Science.gov (United States)

Vasic, Nada; Avrutin, Sergey; Ruigendijk, Esther

2006-01-01

In this paper, we investigate the ability of Dutch agrammatic Broca's and Wernicke's aphasics to assign reference to possessive pronouns in elided VP constructions. The assumption is that the comprehension problems in these two populations have different sources that are revealed in distinct patterns of responses. The focus is primarily on the…

Ultra-high pitch chest computed tomography at 70 kVp tube voltage in an anthropomorphic pediatric phantom and non-sedated pediatric patients: Initial experience with 3rd generation dual-source CT.

Science.gov (United States)

Hagelstein, Claudia; Henzler, Thomas; Haubenreisser, Holger; Meyer, Mathias; Sudarski, Sonja; Schoenberg, Stefan O; Neff, K Wolfgang; Weis, Meike

2016-12-01

Minimizing radiation dose while at the same time preserving image quality is of particular importance in pediatric chest CT. Very recently, CT imaging with a tube voltage of 70 kVp has become clinically available. However, image noise is inversely proportional to the tube voltage. We aimed to investigate radiation dose and image quality of pediatric chest CT performed at 70 kVp in an anthropomorphic pediatric phantom as well as in clinical patients. An anthropomorphic pediatric phantom, which resembles a one-year-old child in physiognomy, was scanned on the 3 rd generation dual-source CT (DSCT) system at 70 kVp and 80 kVp and a fixed ultra low tube-current of 8 mAs to solely evaluate the impact of lowering tube voltage. After the phantom measurements, 18 pediatric patients (mean 29.5 months; range 1-91 months; 21 examinations) underwent 3.2 high-pitch chest CT on the same DSCT system at 70 kVp tube voltage without any sedation. Radiation dose and presence of motion artifacts was compared to a retrospectively identified patient cohort examined at 80 kVp on a 16-slice single-source-CT (SSCT; n=15; 14/15 with sedation; mean 30.7 months; range 0-96 months; pitch=1.5) or on a 2 nd generation DSCT without any sedation (n=6; mean 32.8 months; range 4-61 months; pitch=3.2). Radiation dose in the phantom scans was reduced by approximately 40% when using a tube voltage of 70 kVp instead of 80 kVp. In the pediatric patient group examined at 70 kVp age-specific effective dose (ED; mean 0.5±0.2 mSv) was significantly lower when compared to the retrospective cohort scanned at 80 kVp on the 16-slice-SSCT (mean ED: 1.0±0.3 mSv; pCT examinations showed any motion artifacts whereas 13/15 examinations of the retrospective patient cohort scanned at 80 kVp with a pitch of 1.5 showed motion artifacts. 3.2 high-pitch chest CT performed with 70 kVp significantly reduces radiation dose when compared to 80 kVp while at the same time provides good image quality without any motion artifacts

Principle and engineering implementation of 3D visual representation and indexing of medical diagnostic records (Conference Presentation)

Science.gov (United States)

Shi, Liehang; Sun, Jianyong; Yang, Yuanyuan; Ling, Tonghui; Wang, Mingqing; Zhang, Jianguo

2017-03-01

Purpose: Due to the generation of a large number of electronic imaging diagnostic records (IDR) year after year in a digital hospital, The IDR has become the main component of medical big data which brings huge values to healthcare services, professionals and administration. But a large volume of IDR presented in a hospital also brings new challenges to healthcare professionals and services as there may be too many IDRs for each patient so that it is difficult for a doctor to review all IDR of each patient in a limited appointed time slot. In this presentation, we presented an innovation method which uses an anatomical 3D structure object visually to represent and index historical medical status of each patient, which is called Visual Patient (VP) in this presentation, based on long term archived electronic IDR in a hospital, so that a doctor can quickly learn the historical medical status of the patient, quickly point and retrieve the IDR he or she interested in a limited appointed time slot. Method: The engineering implementation of VP was to build 3D Visual Representation and Index system called VP system (VPS) including components of natural language processing (NLP) for Chinese, Visual Index Creator (VIC), and 3D Visual Rendering Engine.There were three steps in this implementation: (1) an XML-based electronic anatomic structure of human body for each patient was created and used visually to index the all of abstract information of each IDR for each patient; (2)a number of specific designed IDR parsing processors were developed and used to extract various kinds of abstract information of IDRs retrieved from hospital information systems; (3) a 3D anatomic rendering object was introduced visually to represent and display the content of VIO for each patient. Results: The VPS was implemented in a simulated clinical environment including PACS/RIS to show VP instance to doctors. We setup two evaluation scenario in a hospital radiology department to evaluate whether

Enhanced neoplastic transformation by mammography X rays relative to 200 kVp X rays: indication for a strong dependence on photon energy of the RBE(M) for various end points.

Science.gov (United States)

Frankenberg, D; Kelnhofer, K; Bär, K; Frankenberg-Schwager, M

2002-01-01

The fundamental assumption implicit in the use of the atomic bomb survivor data to derive risk estimates is that the gamma rays of Hiroshima and Nagasaki are considered to have biological efficiencies equal to those of other low-LET radiations up to 10 keV/microm, including mammography X rays. Microdosimetric and radiobiological data contradict this assumption. It is therefore of scientific and public interest to evaluate the efficiency of mammography X rays (25-30 kVp) to induce cancer. In this study, the efficiency of mammography X rays relative to 200 kVp X rays to induce neoplastic cell transformation was evaluated using cells of a human hybrid cell line (CGL1). For both radiations, a linear-quadratic dose-effect relationship was observed for neoplastic transformation of CGL1 cells; there was a strong linear component for the 29 kVp X rays. The RBE(M) of mammography X rays relative to 200 kVp X rays was determined to be about 4 for doses energies of transformation of CGL1 cells. Both the data available in the literature and the results of the present study strongly suggest an increase of RBE(M) for carcinogenesis in animals, neoplastic cell transformation, and clastogenic effects with decreasing photon energy or increasing LET to an RBE(M) approximately 8 for mammography X rays relative to 60Co gamma rays.

Multifunctional e-spun colloidal nanofiber structures from various dispersed blends of PVA/ODA-MMT with PVP/ODA-MMT, poly(VP-alt-MA and AgNPs incorporated polymer complexes as electro-active platforms

Directory of Open Access Journals (Sweden)

U. Bunyatova

2016-07-01

Full Text Available This work presented a new approach to fabricate polymer nanocomposites films with nanofiber structures from solution blends of poly(vinyl alcohol + octadecyl amine-montmorillonite (ODA-MMT (matrix with poly(N-vinylpyrrolidone + ODA-MMT (partner-1, poly(N-vinylpyrrolidone-alt-maleic anhydride ((poly(VP-alt-MA + (ODA-MMT (partner-2 and their silver (Ag-carrying polymer complexes by electrospinning. Chemical and physical structures, surface morphologies, thermal behaviors, electrical conductivity and thermal resistance parameters of nanofiber structures were investigated. Poly(VP-alt-MA was used both as a crosslinker and a donor of the hydrophilic groups such as ‒COOH and ‒NH–C=O amide from pyrrolidone ring. Reactive poly(VP-alt-MA, in situ generated Ag nanoparticles (AgNPs and original partner polymer had an significant effect on the morphology and diameter distribution of nanofibers. High and excellent conductive behaviors were observed for the homopolymer and copolymer of VP based fiber structures, respectively. Upon successive chemical cross-linking of the nanofiber structures by reactive partner copolymer, the conductivity of nanofiber films as electro-active platforms dramatically increased to 3.90·10–2 S·cm–1 at room temperature. Comparative analysis results also indicated that electrical properties strongly depended on the loaded reactive organoclay and in situ generated AgNPs.

Routledge History of Women in Modern Europe, 1700 to the present

DEFF Research Database (Denmark)

scholars and rising stars of the discipline to provide a ground-breaking and unique contribution to the historical study of women. Chapters include:European Women's History at the Crossroads, Writing Women in(to) European History, At Home in the Family, Female Sexuality, Learning to be good girls and women......The Routledge History of Women in Europe since 1700 is a landmark publication that provides an over view of women's role and place in Western Europe, spanning three centuries. Drawing on new research and provoking valuable new insights, this comprehensive edited collection brings together renowned......, Women Workers, Working women, Women Religious and religious women, Women as citizens, Women in War and Peace, Home and Away, popular culture and Leisure, Mistresses of creation, women as producers and consumers of culture....

ACTIVITY AND Vp/Vs RATIO OF VOLCANO-TECTONIC SEISMIC SWARM ZONES AT NEVADO DEL RUIZ VOLCANO, COLOMBIA

Directory of Open Access Journals (Sweden)

Londoño B. John Makario

2010-06-01

Full Text Available An analysis of the seismic activity for volcano-tectonic earthquake (VT swarms zones at Nevado del Ruiz Volcano (NRV was carried out for the interval 1985- 2002, which is the most seismic active period at NRV until now (2010. The swarm-like seismicity of NRV was frequently concentrated in very well defined clusters around the volcano. The seismic swarm zone located at the active crater was the most active during the entire time. The seismic swarm zone located to the west of the volcano suggested some relationship with the volcanic crises. It was active before and after the two eruptions occurred in November 1985 and September 1989. It is believed that this seismic activity may be used as a monitoring tool of volcanic activity. For each seismic swarm zone the Vp/Vs ratio was also calculated by grouping of earthquakes and stations. It was found that each seismic swarm zone had a distinct Vp/Vs ratio with respect to the others, except for the crater and west swarm zones, which had the same value. The average Vp/Vs ratios for the seismic swarm zones located at the active crater and to the west of the volcano are about 6-7% lower than that for the north swarm zone, and about 3% lower than that for the south swarm zone. We suggest that the reduction of the Vp/Vs ratio is due to degassing phenomena inside the central and western earthquake swarm zones, or due to the presence of microcracks inside the volcano. This supposition is in agreement with other studies of geophysics, geochemistry and drilling surveys carried out at NRV.

Ultra-high pitch chest computed tomography at 70 kVp tube voltage in an anthropomorphic pediatric phantom and non-sedated pediatric patients. Initial experience with 3{sup rd} generation dual-source CT

Energy Technology Data Exchange (ETDEWEB)

Hagelstein, Claudia; Henzler, Thomas; Haubenreisser, Holger; Meyer, Mathias; Sudarski, Sonja; Schoenberg, Stefan O.; Neff, K. Wolfgang; Weis, Meike [Univ. Medical Center Mannheim (Germany). Inst. of Clinical Radiology and Nuclear Medicine

2016-07-01

Minimizing radiation dose while at the same time preserving image quality is of particular importance in pediatric chest CT. Very recently, CT imaging with a tube voltage of 70 kVp has become clinically available. However, image noise is inversely proportional to the tube voltage. We aimed to investigate radiation dose and image quality of pediatric chest CT performed at 70 kVp in an anthropomorphic pediatric phantom as well as in clinical patients. An anthropomorphic pediatric phantom, which resembles a one-year-old child in physiognomy, was scanned on the 3{sup rd} generation dual-source CT (DSCT) system at 70 kVp and 80 kVp and a fixed ultra low tube-current of 8 mAs to solely evaluate the impact of lowering tube voltage. After the phantom measurements, 18 pediatric patients (mean 29.5 months; range 1-91 months; 21 examinations) underwent 3.2 high-pitch chest CT on the same DSCT system at 70 kVp tube voltage without any sedation. Radiation dose and presence of motion artifacts was compared to a retrospectively identified patient cohort examined at 80 kVp on a 16-slice single-source-CT (SSCT; n = 15; 14/15 with sedation; mean 30.7 months; range 0-96 months; pitch = 1.5) or on a 2{sup nd} generation DSCT without any sedation (n = 6; mean 32.8 months; range 4-61 months; pitch = 3.2). Radiation dose in the phantom scans was reduced by approximately 40% when using a tube voltage of 70 kVp instead of 80 kVp. In the pediatric patient group examined at 70 kVp age-specific effective dose (ED; mean 0.5 ± 0.2 mSv) was significantly lower when compared to the retrospective cohort scanned at 80 kVp on the 16-slice-SSCT (mean ED: 1.0 ± 0.3 mSv; p < 0.0001) and also considerably lower when compared to the cohort scanned at 80 kVp on the 2{sup nd} generation DSCT (mean ED: 0.9 ± 0.5 mSv). None of the prospective, sedation-free CT examinations showed any motion artifacts whereas 13/15 examinations of the retrospective patient cohort scanned at 80 kVp with a pitch of 1

A Sensitive in Vitro High-Throughput Screen To Identify Pan-filoviral Replication Inhibitors Targeting the VP35–NP Interface

Energy Technology Data Exchange (ETDEWEB)

Liu, Gai; Nash, Peter J.; Johnson, Britney; Pietzsch, Colette; Ilagan, Ma. Xenia G.; Bukreyev, Alexander; Basler, Christopher F.; Bowlin, Terry L.; Moir, Donald T.; Leung, Daisy W.; Amarasinghe, Gaya K. (WU-MED); (GSU); (Texas-MED); (Microbiotix)

2017-01-24

The 2014 Ebola outbreak in West Africa, the largest outbreak on record, highlighted the need for novel approaches to therapeutics targeting Ebola virus (EBOV). Within the EBOV replication complex, the interaction between polymerase cofactor, viral protein 35 (VP35), and nucleoprotein (NP) is critical for viral RNA synthesis. We recently identified a peptide at the N-terminus of VP35 (termed NPBP) that is sufficient for interaction with NP and suppresses EBOV replication, suggesting that the NPBP binding pocket can serve as a potential drug target. Here we describe the development and validation of a sensitive high-throughput screen (HTS) using a fluorescence polarization assay. Initial hits from this HTS include the FDA-approved compound tolcapone, whose potency against EBOV infection was validated in a nonfluorescent secondary assay. High conservation of the NP–VP35 interface among filoviruses suggests that this assay has the capacity to identify pan-filoviral inhibitors for development as antivirals.

A Content Analytic Technique for Measuring the Sexiness of Women's Business Attire in Media Presentations.

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White, Sylvia E.

1995-01-01

Describes development of an objective content analytic category scheme for measuring the sexiness of women's business attire in media presentations. Finds women's business attire in television soap operas significantly more provocative than real-world attire. Finds a significant positive correlation between the degree of sexiness as measured by…

pH-induced conformational change of the rotavirus VP4 spike: implications for cell entry and antibody neutralization.

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Pesavento, Joseph B; Crawford, Sue E; Roberts, Ed; Estes, Mary K; Prasad, B V Venkataram

2005-07-01

The rotavirus spike protein, VP4, is a major determinant of infectivity and neutralization. Previously, we have shown that trypsin-enhanced infectivity of rotavirus involves a transformation of the VP4 spike from a flexible to a rigid bilobed structure. Here we show that at elevated pH the spike undergoes a drastic, irreversible conformational change and becomes stunted, with a pronounced trilobed appearance. These particles with altered spikes, at a normal pH of 7.5, despite the loss of infectivity and the ability to hemagglutinate, surprisingly exhibit sialic acid (SA)-independent cell binding in contrast to the SA-dependent cell binding exhibited by native virions. Remarkably, a neutralizing monoclonal antibody that remains bound to spikes throughout the pH changes (pH 7 to 11 and back to pH 7) completely prevents this conformational change, preserving the SA-dependent cell binding and hemagglutinating functions of the virion. A hypothesis that emerges from the present study is that high-pH treatment triggers a conformational change that mimics a post-SA-attachment step to expose an epitope recognized by a downstream receptor in the rotavirus cell entry process. This process involves sequential interactions with multiple receptors, and the mechanism by which the antibody neutralizes is by preventing this conformational change.

Complete protection of cats against feline panleukopenia virus challenge by a recombinant canine adenovirus type 2 expressing VP2 from FPV.

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Yang, Songtao; Xia, Xianzhu; Qiao, Jun; Liu, Quan; Chang, Shuang; Xie, Zhijing; Ju, Huiyan; Zou, Xiaohuan; Gao, Yuwei

2008-03-10

Feline panleukopenia virus (FPV) is an important infectious pathogen of all members of the family Felidae. Here, we describe construction of a replication-competent recombinant canine adenovirus type 2 (CAV-2) expressing the VP2 protein of FPV (CAV-2-VP2) by transfection of MDCK cells with recombinant CAV-2 genome carrying a VP2 expression cassette. Ten 3-month-old cats were vaccinated with the recombinant virus with two boosters at 15-day intervals. All cats developed neutralizing antibodies of titers 1:16-1:32 by day 15 post-primary vaccination, increasing to 1:64-1:128 by day 45. Examination for clinical signs and viral presence, and total white blood cell counts in peripheral blood following FPV challenge, showed that all were completely protected. This recombinant virus appears to provide an effective alternative to attenuated and inactivated vaccines in immunizing cats against feline panleukopenia.

Possible use of CdTe detectors in kVp monitoring of diagnostic X-ray tubes

International Nuclear Information System (INIS)

Krmar, M.; Bucalovic, N.; Baucal, M.; Jovancevic, N.

2010-01-01

It has been suggested that kVp of diagnostic X-ray devices (or maximal energy of X-ray photon spectra) should be monitored routinely; however a standardized non-invasive technique has yet to be developed and proposed. It is well known that the integral number of Compton scattered photons and the intensities of fluorescent X-ray lines registered after irradiation of some material by an X-ray beam are a function of the maximal beam energy. CdTe detectors have sufficient energy resolution to distinguish individual X-ray fluorescence lines and high efficiency for the photon energies in the diagnostic region. Our initial measurements have demonstrated that the different ratios of the integral number of Compton scattered photons and intensities of K and L fluorescent lines detected by CdTe detector are sensitive function of maximal photon energy and could be successfully applied for kVp monitoring.

Role of metabolic activation by cytochrome P-450 in covalent binding of VP 16-213 to rat liver and HeLa cell microsomal proteins

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van Maanen, J.M.; de Ruiter, C.; de Vries, J.; Kootstra, P.R.; Gobas, F.; Pinedo, H.M.

1985-09-01

Covalent binding of /sup 3/H-labeled VP 16-213 to rat liver and HeLa cell microsomal proteins was studied in vitro. Metabolic activation by cytochrome P-450 was found to play a role in the covalent binding of VP 16-213 to rat liver microsomal proteins, as shown by the need of NADPH cofactor, the increased binding after phenobarbital pretreatment and the inhibition by SFK-525A. Addition of ascorbic acid or alpha-phenyl-N-tert. butylnitrone to the incubation mixture depressed covalent binding by about 85%, suggesting that formation of a reactive metabolite from the phenolic structure may be involved in the binding process. VP 16-213 did not inhibit aminopyrine N-demethylase at the concentration used in the binding experiments (17 microM), indicating that metabolism of its methylenedioxy group does not play a role in binding to microsomal proteins. HeLa cell microsomes were found to possess aminopyrine N-demethylase activity. Covalent binding of radiolabeled VP 16-213 to HeLa cell microsomes decreased by about 64% if NADPH was omitted.

Identification of mud crab reovirus VP12 and its interaction with the voltage-dependent anion-selective channel protein of mud crab Scylla paramamosain.

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Xu, Hai-Dong; Su, Hong-Jun; Zou, Wei-Bin; Liu, Shan-Shan; Yan, Wen-Rui; Wang, Qian-Qian; Yuan, Li-Li; Chan, Siuming Francis; Yu, Xiao-Qiang; He, Jian-Guo; Weng, Shao-Ping

2015-05-01

Mud crab reovirus (MCRV) is the causative agent of a severe disease in cultured mud crab (Scylla paramamosain), which has caused huge economic losses in China. MCRV is a double-stranded RNA virus with 12 genomic segments. In this paper, SDS-PAGE, mass spectrometry and Western blot analyses revealed that the VP12 protein encoded by S12 gene is a structural protein of MCRV. Immune electron microscopy assay indicated that MCRV VP12 is a component of MCRV outer shell capsid. Yeast two hybrid cDNA library of mud crab was constructed and mud crab voltage-dependent anion-selective channel (mcVDAC) was obtained by MCRV VP12 screening. The full length of mcVDAC was 1180 bp with an open reading frame (ORF) of 849 bp encoding a 282 amino acid protein. The mcVDAC had a constitutive expression pattern in different tissues of mud crab. The interaction between MCRV VP12 and mcVDAC was determined by co-immunoprecipitation assay. The results of this study have provided an insight on the mechanisms of MCRV infection and the interactions between the virus and mud crab. Copyright © 2015 Elsevier Ltd. All rights reserved.

Detection of polyomavirus major capsid antigen (VP-1 in human pilomatricomas Detección del antígeno mayor de la cápside de poliomavirus (VP-1 en pilomatricomas humanos

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Norberto A. Sanjuán

2010-04-01

Full Text Available The family Polyomaviridae is composed of small, non-enveloped, double-stranded DNA viruses widely used to study cell transformation in vitro and tumor induction in vivo. The development of pilomatricomas in mice experimentally infected with polyomavirus led us to detect the viral major capsid protein VP-1 in human pilomatricomas. This tumor, even uncommon, is one of the most frequent benign hair follicle tumors in humans and is composed of proliferating matrix cells that undergo keratinization, and form cystic neoplasms. The detection of VP-1 was performed using the peroxidase-antiperoxidase technique in paraffin-embedded slides with a specific primary serum. Adjacent slides treated with normal rabbit serum as a primary were employed as internal control. Positive and negative controls were also employed as well as slides of lesions caused by human papillomavirus to rule out any unspecific cross-reactivity. In 4 out of 10 cases polyomavirus VP-1 was clearly detected in nuclei of human pilomatricomas proliferating cells, in a patchy pattern of distribution. The controls confirmed the specificity of the immunocytochemical procedure. These results could indicate either an eventual infection of the virus in already developed tumors or alternatively, a direct involvement of polyomavirus in the pathogenesis of some pilomatricomas. The recent discovery of a new human polyomavirus associated with Merkel cell carcinomas has been a strong contribution to better understand the pathogenesis of some human uncommon skin cancers. Hopefully the results reported in this work will encourage further research on the role of polyomavirus in other human skin neoplasms.La familia Poliomaviridae está compuesta por virus oncogénicos pequeños, no envueltos, con ADN de doble cadena. En un modelo experimental murino pudimos desarrollar pilomatricomas inducidos por la inoculación de virus polioma. Eso nos llevó a estudiar la posibilidad de que otro virus polioma

Using 80 kVp on a 320-row scanner for hepatic multiphasic CT reduces the contrast dose by 50 % in patients at risk for contrast-induced nephropathy

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Taguchi, Narumi; Oda, Seitaro; Utsunomiya, Daisuke; Nakaura, Takeshi; Imuta, Masanori; Yamamura, Sadahiro; Yuki, Hideaki; Kidoh, Masafumi; Hirata, Kenichiro; Namimoto, Tomohiro; Yamashita, Yasuyuki [Kumamoto University, Department of Diagnostic Radiology, Faculty of Life Sciences, Kumamoto (Japan); Funama, Yoshinori [Kumamoto University, Department of Medical Physics, Faculty of Life Sciences, Kumamoto (Japan); Hatemura, Masahiro; Kai, Noriyuki [Kumamoto University Hospital, Department of Central Radiology, Kumamoto (Japan)

2017-02-15

We evaluated the effects of a low contrast material (CM) dose protocol using 80-kVp on the image quality of hepatic multiphasic CT scans acquired on a 320-row CT scanner. We scanned 30 patients with renal insufficiency (eGFR < 45 mL/min/1.73 m{sup 2}) using 80-kVp and a CM dose of 300mgI/kg. Another 30 patients without renal insufficiency (eGFR > 60 mL/min/1.73 m{sup 2}) were scanned with the conventional 120-kVp protocol and the standard CM dose of 600mgI/kg. Quantitative image quality parameters, i.e. CT attenuation, image noise, and the contrast-to-noise ratio (CNR) were compared and the visual image quality was scored on a four-point scale. The volume CT dose index (CTDI{sub vol}) and the size-specific dose estimate (SSDE) recorded with the 80- and the 120-kVp protocols were also compared. Image noise and contrast enhancement were equivalent for the two protocols. There was no significant difference in the CNR of all anatomic sites and in the visual scores for overall image quality. The CTDI{sub vol} and SSDE were approximately 25-30 % lower under the 80-kVp protocol. Hepatic multiphase CT using 80-kVp on a 320-row CT scanner allowed for a decrease in the CM dose and a reduction in the radiation dose without image quality degradation in patients with renal insufficiency. (orig.)

Promoting Early Presentation of Breast Cancer in Older Women: Implementing an Evidence-Based Intervention in Routine Clinical Practice

International Nuclear Information System (INIS)

Forbes, L. J. L.; Forster, A. S.; Dodd, R. H.; Tucker, L.; Laming, R.; Ramirez, A. J.; Sellars, S.; Patnick, J.

2012-01-01

Women over 70 with breast cancer have poorer one-year survival and present at a more advanced stage than younger women. Promoting early symptomatic presentation in older women may reduce stage cost effectively and is unlikely to lead to overdiagnosis. After examining efficacy in a randomised controlled trial, we piloted a brief health professional-delivered intervention to equip women to present promptly with breast symptoms, as an integral part of the final invited mammogram at age ∼70, in the English National Health Service Breast Screening Programme. Methods. We trained mammographers, who then offered the intervention to older women in four breast screening services. We examined breast cancer awareness at baseline and one month in women receiving the intervention, and also in a service where the intervention was not offered. Results. We trained 27 mammographers to deliver the intervention confidently to a high standard. Breast cancer awareness increased 7-fold at one month in women receiving the intervention compared with 2-fold in the comparison service (odds ratio 15.2, 95% confidence interval 10.0 to 23.2). Conclusions. The PEP Intervention can be implemented in routine clinical practice with a potency similar to that achieved in a randomised controlled trial. It has the potential to reduce delay in diagnosis for breast cancer in older women.

Promoting Early Presentation of Breast Cancer in Older Women: Implementing an Evidence-Based Intervention in Routine Clinical Practice

Science.gov (United States)

Forbes, Lindsay J. L.; Forster, Alice S.; Dodd, Rachael H.; Tucker, Lorraine; Laming, Rachel; Sellars, Sarah; Patnick, Julietta; Ramirez, Amanda J.

2012-01-01

Background. Women over 70 with breast cancer have poorer one-year survival and present at a more advanced stage than younger women. Promoting early symptomatic presentation in older women may reduce stage cost effectively and is unlikely to lead to overdiagnosis. After examining efficacy in a randomised controlled trial, we piloted a brief health professional-delivered intervention to equip women to present promptly with breast symptoms, as an integral part of the final invited mammogram at age ~70, in the English National Health Service Breast Screening Programme. Methods. We trained mammographers, who then offered the intervention to older women in four breast screening services. We examined breast cancer awareness at baseline and one month in women receiving the intervention, and also in a service where the intervention was not offered. Results. We trained 27 mammographers to deliver the intervention confidently to a high standard. Breast cancer awareness increased 7-fold at one month in women receiving the intervention compared with 2-fold in the comparison service (odds ratio 15.2, 95% confidence interval 10.0 to 23.2). Conclusions. The PEP Intervention can be implemented in routine clinical practice with a potency similar to that achieved in a randomised controlled trial. It has the potential to reduce delay in diagnosis for breast cancer in older women. PMID:23213334

A proposed vestigial translation initiation motif in VP1 of hepatitis A virus.

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Kang, Jeong-Ah; Funkhouser, Ann W

2002-07-01

The internal ribosome entry site (IRES) of picornaviruses has a 3' polypyrimidine tract (PPT) 16-24 bases upstream of an AUG triplet (PPT/AUG motif). This motif is critical in determining the efficiency of cap-independent translation. HAV has a conserved PPT/AUG motif consisting of a nine base sequence (AGGUUUUUC) 23 bases upstream of the preferred AUG start codon. This HAV-specific PPT/AUG motif is repeated and conserved in VP1 of HAV, but not of other picornaviruses. We proposed that the PPT/AUG motif in the open reading frame initiated translation and/or had an impact on the life cycle of the virus. In vitro translation of mutant bicistronic mRNAs and growth in cell culture of mutant viruses provided no evidence that the VP1 PPT/AUG motif had any impact on either translation or growth. HAV differs from other picornaviruses in its inefficient growth in cell culture. Since the HAV-specific PPT/AUG motif is found in only 1 in 300,000 reported viral sequences outside the hepatovirus genus, this motif may be a vestigial translation initiation element and may have played a role in determining the unusual phenotype of HAV.

Self-presentation in Online Professional Networks: Men's Higher and Women's Lower Facial Prominence in Self-created Profile Images.

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Sczesny, Sabine; Kaufmann, Michèle C

2017-01-01

Men are presented with higher facial prominence than women in the media, a phenomenon that is called face-ism . In naturalistic settings, face-ism effects could be driven by gender biases of photographers and/or by gender differences in self-presentation. The present research is the first to investigate whether women and men themselves create this different facial prominence. In a controlled laboratory study, 61 participants prepared a picture of themselves from a half-body photograph, allegedly to be uploaded to their profile for an online professional network. As expected, men cropped their photos with higher facial prominence than women did. However, women and men did not differ in the self-presentational motivations, goals, strategies, and personality variables under investigation, so that the observed face-ism effect could not be explained with these variables. Generally, the higher participants' physical appearance self-esteem, the higher was their self-created facial prominence.

Experimental article – Reducing effective dose to a paediatric phantom by using different combinations of kVp, mAs and additional filtration whilst maintaining image quality

NARCIS (Netherlands)

Elsbeth Huizinga; Ana Pereira; Wouter Schaake; Charlotte Bloomfield; Heidi Knight; Emilie Crausaz; Hanne Hustveit; Ruurd Visser; Vanja Harsaker; Diane Chabloz; Filipa Boavida

2015-01-01

Purpose: To determine whether using different combinations of kVp and mAs with additional filtration can reduce the effective dose to a paediatric phantom whilst maintaining diagnostic image quality. Methods: 27 images of a paediatric AP pelvis phantom were acquired with different kVp, mAs

Evaluation of a decision aid for women with breech presentation at term: a randomised controlled trial [ISRCTN14570598

Science.gov (United States)

Nassar, N; Roberts, CL; Raynes-Greenow, CH; Barratt, A; Peat, B

2007-01-01

Objectives To evaluate the effectiveness of a decision aid for women with a breech presentation compared with usual care. Design Randomised controlled trial. Setting Tertiary obstetric hospitals offering external cephalic version (ECV). Population Women with a singleton pregnancy were diagnosed antenatally with a breech presentation at term, and were clinically eligible for ECV. Methods Women were randomised to either receive a decision aid about the management options for breech presentation in addition to usual care or to receive usual care only with standard counselling from their usual pregnancy care provider. The decision aid comprised a 24-page booklet supplemented by a 30-minute audio-CD and worksheet that was designed for women to take home and review with a partner. Main outcome measures Decisional conflict (uncertainty), knowledge, anxiety and satisfaction with decision making, and were assessed using self-administered questionnaires. Results Compared with usual care, women reviewing the decision aid experienced significantly lower decisional conflict (mean difference −8.92; 95% CI −13.18, −4.66) and increased knowledge (mean difference 8.40; 95% CI 3.10, 13.71), were more likely to feel that they had enough information to make a decision (RR 1.30; 95% CI 1.14, 1.47), had no increase in anxiety and reported greater satisfaction with decision making and overall experience of pregnancy and childbirth. In contrast, 19% of women in the usual care group reported they would have made a different decision about their care. Conclusions A decision aid is an effective and acceptable tool for pregnant women that provides an important adjunct to standard counselling for the management of breech presentation. Please cite this paper as: Nassar N, Roberts C, Raynes-Greenow C, Barratt A, Peat B, on behalf of the Decision Aid for Breech Presentation Trial Collaborators. Evaluation of a decision aid for women with breech presentation at term: a randomised controlled

Spatiotemporal Phylogenetic Analysis and Molecular Characterisation of Infectious Bursal Disease Viruses Based on the VP2 Hyper-Variable Region.

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Abdulahi Alfonso-Morales

Full Text Available Infectious bursal disease is a highly contagious and acute viral disease caused by the infectious bursal disease virus (IBDV; it affects all major poultry producing areas of the world. The current study was designed to rigorously measure the global phylogeographic dynamics of IBDV strains to gain insight into viral population expansion as well as the emergence, spread and pattern of the geographical structure of very virulent IBDV (vvIBDV strains.Sequences of the hyper-variable region of the VP2 (HVR-VP2 gene from IBDV strains isolated from diverse geographic locations were obtained from the GenBank database; Cuban sequences were obtained in the current work. All sequences were analysed by Bayesian phylogeographic analysis, implemented in the Bayesian Evolutionary Analysis Sampling Trees (BEAST, Bayesian Tip-association Significance testing (BaTS and Spatial Phylogenetic Reconstruction of Evolutionary Dynamics (SPREAD software packages. Selection pressure on the HVR-VP2 was also assessed. The phylogeographic association-trait analysis showed that viruses sampled from individual countries tend to cluster together, suggesting a geographic pattern for IBDV strains. Spatial analysis from this study revealed that strains carrying sequences that were linked to increased virulence of IBDV appeared in Iran in 1981 and spread to Western Europe (Belgium in 1987, Africa (Egypt around 1990, East Asia (China and Japan in 1993, the Caribbean Region (Cuba by 1995 and South America (Brazil around 2000. Selection pressure analysis showed that several codons in the HVR-VP2 region were under purifying selection.To our knowledge, this work is the first study applying the Bayesian phylogeographic reconstruction approach to analyse the emergence and spread of vvIBDV strains worldwide.

Spatiotemporal Phylogenetic Analysis and Molecular Characterisation of Infectious Bursal Disease Viruses Based on the VP2 Hyper-Variable Region.

Science.gov (United States)

Alfonso-Morales, Abdulahi; Martínez-Pérez, Orlando; Dolz, Roser; Valle, Rosa; Perera, Carmen L; Bertran, Kateri; Frías, Maria T; Majó, Natàlia; Ganges, Llilianne; Pérez, Lester J

2013-01-01

Infectious bursal disease is a highly contagious and acute viral disease caused by the infectious bursal disease virus (IBDV); it affects all major poultry producing areas of the world. The current study was designed to rigorously measure the global phylogeographic dynamics of IBDV strains to gain insight into viral population expansion as well as the emergence, spread and pattern of the geographical structure of very virulent IBDV (vvIBDV) strains. Sequences of the hyper-variable region of the VP2 (HVR-VP2) gene from IBDV strains isolated from diverse geographic locations were obtained from the GenBank database; Cuban sequences were obtained in the current work. All sequences were analysed by Bayesian phylogeographic analysis, implemented in the Bayesian Evolutionary Analysis Sampling Trees (BEAST), Bayesian Tip-association Significance testing (BaTS) and Spatial Phylogenetic Reconstruction of Evolutionary Dynamics (SPREAD) software packages. Selection pressure on the HVR-VP2 was also assessed. The phylogeographic association-trait analysis showed that viruses sampled from individual countries tend to cluster together, suggesting a geographic pattern for IBDV strains. Spatial analysis from this study revealed that strains carrying sequences that were linked to increased virulence of IBDV appeared in Iran in 1981 and spread to Western Europe (Belgium) in 1987, Africa (Egypt) around 1990, East Asia (China and Japan) in 1993, the Caribbean Region (Cuba) by 1995 and South America (Brazil) around 2000. Selection pressure analysis showed that several codons in the HVR-VP2 region were under purifying selection. To our knowledge, this work is the first study applying the Bayesian phylogeographic reconstruction approach to analyse the emergence and spread of vvIBDV strains worldwide.

Quantitative statistical analysis of cis-regulatory sequences in ABA/VP1- and CBF/DREB1-regulated genes of Arabidopsis.

Science.gov (United States)

Suzuki, Masaharu; Ketterling, Matthew G; McCarty, Donald R

2005-09-01

We have developed a simple quantitative computational approach for objective analysis of cis-regulatory sequences in promoters of coregulated genes. The program, designated MotifFinder, identifies oligo sequences that are overrepresented in promoters of coregulated genes. We used this approach to analyze promoter sequences of Viviparous1 (VP1)/abscisic acid (ABA)-regulated genes and cold-regulated genes, respectively, of Arabidopsis (Arabidopsis thaliana). We detected significantly enriched sequences in up-regulated genes but not in down-regulated genes. This result suggests that gene activation but not repression is mediated by specific and common sequence elements in promoters. The enriched motifs include several known cis-regulatory sequences as well as previously unidentified motifs. With respect to known cis-elements, we dissected the flanking nucleotides of the core sequences of Sph element, ABA response elements (ABREs), and the C repeat/dehydration-responsive element. This analysis identified the motif variants that may correlate with qualitative and quantitative differences in gene expression. While both VP1 and cold responses are mediated in part by ABA signaling via ABREs, these responses correlate with unique ABRE variants distinguished by nucleotides flanking the ACGT core. ABRE and Sph motifs are tightly associated uniquely in the coregulated set of genes showing a strict dependence on VP1 and ABA signaling. Finally, analysis of distribution of the enriched sequences revealed a striking concentration of enriched motifs in a proximal 200-base region of VP1/ABA and cold-regulated promoters. Overall, each class of coregulated genes possesses a discrete set of the enriched motifs with unique distributions in their promoters that may account for the specificity of gene regulation.

Phylogenetic reconstruction and polymorphism analysis of BK virus VP2 gene isolated from renal transplant recipients in China.

Science.gov (United States)

Wang, Zhang-Yang; Hong, Wei-Long; Zhu, Zhe-Hui; Chen, Yun-Hao; Ye, Wen-LE; Chu, Guang-Yu; Li, Jia-Lin; Chen, Bi-Cheng; Xia, Peng

2015-11-01

BK polyomavirus (BKV) is important pathogen for kidney transplant recipients, as it is frequently re-activated, leading to nephropathy. The aim of this study was to investigate the phylogenetic reconstruction and polymorphism of the VP2 gene in BKV isolated from Chinese kidney transplant recipients. Phylogenetic analysis was carried out in the VP2 region from 135 BKV-positive samples and 28 reference strains retrieved from GenBank. The unweighted pair-group method with arithmetic mean (UPGMA) grouped all strains into subtypes, but failed to subdivide strains into subgroups. Among the plasma and urine samples, all plasma (23/23) and 82 urine samples (82/95) were identified to contain subtype I; the other 10 urine samples contained subtype IV. A 86-bp fragment was identified as a highly conserved sequence. Following alignment with 36 published BKV sequences from China, 92 sites of polymorphism were identified, including 11 single nucleotide polymorphisms (SNPs) prevalent in Chinese individuals and 30 SNPs that were specific to the two predominant subtypes I and IV. The limitations of the VP2 gene segment in subgrouping were confirmed by phylogenetic analysis. The conserved sequence and polymorphism identified in this study may be helpful in the detection and genotyping of BKV.

Rhinovirus-induced VP1-specific Antibodies are Group-specific and Associated With Severity of Respiratory Symptoms

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Katarzyna Niespodziana

2015-01-01

Interpretation: Our results demonstrate that increases of antibodies towards the VP1 N-terminus are group-specific and associated with severity of respiratory symptoms and suggest that it may be possible to develop serological tests for identifying causative RV groups.

De Novo Herpes Simplex Virus VP16 Expression Gates a Dynamic Programmatic Transition and Sets the Latent/Lytic Balance during Acute Infection in Trigeminal Ganglia.

Science.gov (United States)

Sawtell, Nancy M; Thompson, Richard L

2016-09-01

The life long relationship between herpes simplex virus and its host hinges on the ability of the virus to aggressively replicate in epithelial cells at the site of infection and transport into the nervous system through axons innervating the infection site. Interaction between the virus and the sensory neuron represents a pivot point where largely unknown mechanisms lead to a latent or a lytic infection in the neuron. Regulation at this pivot point is critical for balancing two objectives, efficient widespread seeding of the nervous system and host survival. By combining genetic and in vivo in approaches, our studies reveal that the balance between latent and lytic programs is a process occurring early in the trigeminal ganglion. Unexpectedly, activation of the latent program precedes entry into the lytic program by 12 -14hrs. Importantly, at the individual neuronal level, the lytic program begins as a transition out of this acute stage latent program and this escape from the default latent program is regulated by de novo VP16 expression. Our findings support a model in which regulated de novo VP16 expression in the neuron mediates entry into the lytic cycle during the earliest stages of virus infection in vivo. These findings support the hypothesis that the loose association of VP16 with the viral tegument combined with sensory axon length and transport mechanisms serve to limit arrival of virion associated VP16 into neuronal nuclei favoring latency. Further, our findings point to specialized features of the VP16 promoter that control the de novo expression of VP16 in neurons and this regulation is a key component in setting the balance between lytic and latent infections in the nervous system.

Experimental article – Reducing effective dose to a paediatric phantom by using different combinations of kVp, mAs and additional filtration whilst maintaining image quality

NARCIS (Netherlands)

Huizinga, Elsbeth; Schaake, Wouter; Visser, Ruurd; Bloomfield, Charlotte; Boavida, Filipa; Chabloz, Diane; Crausaz, Emilie; Hustveit, Hanne; Knight, Heidi; Pereira, Anna; Harsaker, Vanja

2015-01-01

Purpose: To determine whether using different combinations of kVp and mAs with additional filtration can reduce the effective dose to a paediatric phantom whilst maintaining diagnostic image quality. Methods: 27 images of a paediatric AP pelvis phantom were acquired with different kVp, mAs and

Natural type 3/type 2 intertypic vaccine-related poliovirus recombinants with the first crossover sites within the VP1 capsid coding region.

Science.gov (United States)

Zhang, Yong; Zhu, Shuangli; Yan, Dongmei; Liu, Guiyan; Bai, Ruyin; Wang, Dongyan; Chen, Li; Zhu, Hui; An, Hongqiu; Kew, Olen; Xu, Wenbo

2010-12-21

Ten uncommon natural type 3/type 2 intertypic poliovirus recombinants were isolated from stool specimens from nine acute flaccid paralysis case patients and one healthy vaccinee in China from 2001 to 2008. Complete genomic sequences revealed their vaccine-related genomic features and showed that their first crossover sites were randomly distributed in the 3' end of the VP1 coding region. The length of donor Sabin 2 sequences ranged from 55 to 136 nucleotides, which is the longest donor sequence reported in the literature for this type of poliovirus recombination. The recombination resulted in the introduction of Sabin 2 neutralizing antigenic site 3a (NAg3a) into a Sabin 3 genomic background in the VP1 coding region, which may have been altered by some of the type 3-specific antigenic properties, but had not acquired any type 2-specific characterizations. NAg3a of the Sabin 3 strain seems atypical; other wild-type poliovirus isolates that have circulated in recent years have sequences of NAg3a more like the Sabin 2 strain. 10 natural type 3/type 2 intertypic VP1 capsid-recombinant polioviruses, in which the first crossover sites were found to be in the VP1 coding region, were isolated and characterized. In spite of the complete replacement of NAg3a by type 2-specific amino acids, the serotypes of the recombinants were not altered, and they were totally neutralized by polyclonal type 3 antisera but not at all by type 2 antisera. It is possible that recent type 3 wild poliovirus isolates may be a recombinant having NAg3a sequences derived from another strain during between 1967 and 1980, and the type 3/type 2 recombination events in the 3' end of the VP1 coding region may result in a higher fitness.

Natural type 3/type 2 intertypic vaccine-related poliovirus recombinants with the first crossover sites within the VP1 capsid coding region.

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Yong Zhang

Full Text Available BACKGROUND: Ten uncommon natural type 3/type 2 intertypic poliovirus recombinants were isolated from stool specimens from nine acute flaccid paralysis case patients and one healthy vaccinee in China from 2001 to 2008. PRINCIPAL FINDINGS: Complete genomic sequences revealed their vaccine-related genomic features and showed that their first crossover sites were randomly distributed in the 3' end of the VP1 coding region. The length of donor Sabin 2 sequences ranged from 55 to 136 nucleotides, which is the longest donor sequence reported in the literature for this type of poliovirus recombination. The recombination resulted in the introduction of Sabin 2 neutralizing antigenic site 3a (NAg3a into a Sabin 3 genomic background in the VP1 coding region, which may have been altered by some of the type 3-specific antigenic properties, but had not acquired any type 2-specific characterizations. NAg3a of the Sabin 3 strain seems atypical; other wild-type poliovirus isolates that have circulated in recent years have sequences of NAg3a more like the Sabin 2 strain. CONCLUSIONS: 10 natural type 3/type 2 intertypic VP1 capsid-recombinant polioviruses, in which the first crossover sites were found to be in the VP1 coding region, were isolated and characterized. In spite of the complete replacement of NAg3a by type 2-specific amino acids, the serotypes of the recombinants were not altered, and they were totally neutralized by polyclonal type 3 antisera but not at all by type 2 antisera. It is possible that recent type 3 wild poliovirus isolates may be a recombinant having NAg3a sequences derived from another strain during between 1967 and 1980, and the type 3/type 2 recombination events in the 3' end of the VP1 coding region may result in a higher fitness.

Self-presentation in Online Professional Networks: Men's Higher and Women's Lower Facial Prominence in Self-created Profile Images

Science.gov (United States)

Sczesny, Sabine; Kaufmann, Michèle C.

2018-01-01

Men are presented with higher facial prominence than women in the media, a phenomenon that is called face-ism. In naturalistic settings, face-ism effects could be driven by gender biases of photographers and/or by gender differences in self-presentation. The present research is the first to investigate whether women and men themselves create this different facial prominence. In a controlled laboratory study, 61 participants prepared a picture of themselves from a half-body photograph, allegedly to be uploaded to their profile for an online professional network. As expected, men cropped their photos with higher facial prominence than women did. However, women and men did not differ in the self-presentational motivations, goals, strategies, and personality variables under investigation, so that the observed face-ism effect could not be explained with these variables. Generally, the higher participants' physical appearance self-esteem, the higher was their self-created facial prominence. PMID:29387029

Self-presentation in Online Professional Networks: Men's Higher and Women's Lower Facial Prominence in Self-created Profile Images

Directory of Open Access Journals (Sweden)

Sabine Sczesny

2018-01-01

Full Text Available Men are presented with higher facial prominence than women in the media, a phenomenon that is called face-ism. In naturalistic settings, face-ism effects could be driven by gender biases of photographers and/or by gender differences in self-presentation. The present research is the first to investigate whether women and men themselves create this different facial prominence. In a controlled laboratory study, 61 participants prepared a picture of themselves from a half-body photograph, allegedly to be uploaded to their profile for an online professional network. As expected, men cropped their photos with higher facial prominence than women did. However, women and men did not differ in the self-presentational motivations, goals, strategies, and personality variables under investigation, so that the observed face-ism effect could not be explained with these variables. Generally, the higher participants' physical appearance self-esteem, the higher was their self-created facial prominence.

Kinetics of oxidation of the alloy-MR-47VP with nitrogen dioxide

International Nuclear Information System (INIS)

Vasil'eva, A.G.; Rakova, N.N.; Vladimirskaya, I.N.; Kabanova, O.V.; Miklyaev, A.D.

1978-01-01

The kinetic dependences of oxidation of MR-47VP grade molybdenum-rhenium alloy with nitrogen dioxide have been examined within the temperature range of 350 to 550 deg C. It has been shown that the processes take place in the transition region. The specific oxidation rate of the alloy with the nitrogen dioxide is but small, and it is comparable as to its value with the specific rate of its oxidation in oxygen under identical conditions

New methodology to measure kVp in X-rays equipment for radiodiagnosis

International Nuclear Information System (INIS)

Vega Ramirez, J.L.; Pela, C.A.

1998-01-01

The work aims to obtain through experiments a spaced out sensibilization (different optical densities) in the X-rays film by letting X-rays photons go through a metallic layer of copper with tiers with different thickness that can cover the range of kVps employed in radiological tests. Results for the kVp shown here were also compared with other equipment available in the market. Comparisons showed their similarity but have also the advantage of being cheap and viable

Perception of Labour Pain among Rural Women Presenting to A ...

African Journals Online (AJOL)

Background: Childbirth results in severe pain for many women. In many hospitals in Resource-Limited Countries (RLCs), women endure the pain of labour with little or no pain relief. There have not been any studies done within 36 hours of a recent delivery to determine how rural Kenyan women perceive the pain of labour, ...

Herpes simplex virus type 1 tegument protein VP22 interacts with TAF-I proteins and inhibits nucleosome assembly but not regulation of histone acetylation by INHAT.

Science.gov (United States)

van Leeuwen, Hans; Okuwaki, Mitsuru; Hong, Rui; Chakravarti, Debabrata; Nagata, Kyosuke; O'Hare, Peter

2003-09-01

Affinity chromatography was used to identify cellular proteins that interact with the herpes simplex virus (HSV) tegument protein VP22. Among a small set of proteins that bind specifically to VP22, we identified TAF-I (template-activating factor I), a chromatin remodelling protein and close homologue of the histone chaperone protein NAP-1. TAF-I has been shown previously to promote more ordered transfer of histones to naked DNA through a direct interaction with histones. TAF-I, as a subunit of the INHAT (inhibitor of acetyltransferases) protein complex, also binds to histones and masks them from being substrates for the acetyltransferases p300 and PCAF. Using in vitro assays for TAF-I activity in chromatin assembly, we show that VP22 inhibits nucleosome deposition on DNA by binding to TAF-I. We also observed that VP22 binds non-specifically to DNA, an activity that is abolished by TAF-I. However, the presence of VP22 does not affect the property of INHAT in inhibiting the histone acetyltransferase activity of p300 or PCAF in vitro. We speculate that this interaction could be relevant to HSV DNA organization early in infection, for example, by interfering with nucleosomal deposition on the genome. Consistent with this possibility was the observation that overexpression of TAF-I in transfected cells interferes with the progression of HSV-1 infection.

Barriers to early presentation and diagnosis of breast cancer among African women living in sub-Saharan Africa.

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Cynthia Pomaa Akuoko

Full Text Available Breast cancer (BC has been described as the leading cause of cancer deaths among women especially in the developing world including sub Saharan Africa (SSA. Delayed presentation and late diagnosis at health facilities are parts of the contributing factors of high BC mortality in Africa. This review aimed to appraise the contributing factors to delayed breast cancer presentation and diagnosis among SSA women.Five databases encompassing medical and social sciences were systematically searched using predefined search terms linked with breast cancer presentation and diagnosis and sub Saharan Africa. Reference lists of relevant papers were also hand searched. Quality of quantitative and qualitative articles were assessed using the National Institute of Health (NIH Quality Assessment Tool for Observational Cohort and Cross-Sectional Studies and the Critical Appraisal Skills Programme (CASP quality appraisal checklist. Thematic analysis was used to synthesize the qualitative studies to integrate findings.Fourteen (14 quantitative studies, two (2 qualitative studies and one (1 mixed method study merited inclusion for analysis. This review identified low knowledge of breast cancer among SSA women. This review also found lack of awareness of early detection treatment, poor perception of BC, socio-cultural factors such as belief, traditions and fear as factors impacting African women's health seeking behavior in relation to breast cancer.Improving African women's knowledge and understanding will improve behaviors related to breast cancer and facilitate early presentation and detection and enhance proper management and treatment of breast cancer.

Women of the Solar Physics Division

Science.gov (United States)

Dupree, Andrea K.

2007-05-01

In 1970, when the Solar Physics Division was established, the invitation to become a founding member of the Division was extended by the Organizing Committee to a group of 61 solar scientists of which 4 were women (6.6%). At the first SPD meeting in Huntsville AL (1970), 11% of the papers were given by women. Near that time (1973), women accounted for 8% of all AAS members. The representation of women in the SPD has more than doubled in percentage since the first years. Currently, women comprise about 15.5% of SPD members which, however, is less than the percentage in the AAS general membership (18%) in March 2007. In the 37 years that the SPD has existed, women have frequently held the office of Treasurer and Secretary of the Division and made notable contributions. Elske V.P. Smith was elected the first Treasurer of the SPD and that began a long tradition. Women appear to be considered exceptionally trustworthy since they have been reelected and occupied the position of Treasurer for 75% of the available terms. The Office of SPD Secretary has seen a woman for 13% of the terms. Yet women are practically absent among those in the top leadership positions and in the lists of prize winners of the SPD. Among the 21 SPD Chairs, only 1 woman, Judith T. Karpen, has held that office. The Hale Prize has been awarded 19 times in almost 3 decades, and all of the awardees have been men. Several aspects of the participation of women and their contributions to the Solar Physics Division of the AAS will be reviewed, and compared to that of the AAS and astronomy in general.

Study to Evaluate Targeted Management and Syndromic Management in Women Presenting with Abnormal Vaginal Discharge.

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Meena, Veena; Bansal, Charu Lata

2016-10-01

Vaginal discharge is a commonest complaint among women in reproductive age group. Infective vaginal discharge can be broadly categorized into vaginitis or mucopurulent cervicitis. Vaginitis is predominantly caused by bacterial vaginosis, vaginal candidiasis, vaginal trichomoniasis, etc. Mucopurulent cervicitis is due to chlamydia or gonococcal infection. The targeted management is based on identification of causative organism and targeting the therapy against it, while syndromic management is based on high risk factors's presence. To study the effect of targeted management as compared to syndromic management in achieving a complete cure for abnormal vaginal discharge and to study the microbial flora of women presenting with abnormal vaginal discharge. The study is a randomized control trial conducted at tertiary health care on 200 women who presented with abnormal vaginal discharge, distributed in two groups A and B each consisted of 100 women. Group A underwent laboratory investigations, and treatment was started as soon as reports were available. Group B was given syndromic management based on high risk factors's presence. Both groups were followed up after 2 weeks. The prevalence of various organisms in vaginal discharge was candidiasis 39 %, bacterial vaginosis 28 %, trichomoniasis 5 %, N. gonorrhoeae 5 % and chlamydia 2 % among the 100 women in group A. Response to treatment for vaginitis was 76.3 % in group A, whereas it was 41 % in group B. With cervicitis, 71.4 % women responded to treatment in targeted group as compared to 54 % in syndromic management group. There is a potential disadvantage of syndromic management because of its total reliability on a subjective clinical assessment.

Antigenic heterogeneity of capsid protein VP1 in foot-and-mouth disease virus (FMDV serotype Asia1

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Alam SM

2013-08-01

Full Text Available SM Sabbir Alam,1 Ruhul Amin,1 Mohammed Ziaur Rahman,2 M Anwar Hossain,1 Munawar Sultana11Department of Microbiology, University of Dhaka, Dhaka, Bangladesh; 2International Centre for Diarrhoeal Disease Research, Dhaka, BangladeshAbstract: Foot and mouth disease virus (FMDV, with its seven serotypes, is a highly contagious virus infecting mainly cloven-hoofed animals. The serotype Asia1 occurs mainly in Asian regions. An in-silico approach was taken to reveal the antigenic heterogeneities within the capsid protein VP1 of Asia1. A total of 47 VP1 sequences of Asia1 isolates from different countries of South Asian regions were selected, retrieved from database, and were aligned. The structure of VP1 protein was modeled using a homology modeling approach. Several antigenic sites were identified and mapped onto the three-dimensional protein structure. Variations at these antigenic sites were analyzed by calculating the protein variability index and finding mutation combinations. The data suggested that vaccine escape mutants have derived from only few mutations at several antigenic sites. Five antigenic peptides have been identified as the least variable epitopes, with just fewer amino acid substitutions. Only a limited number of serotype Asia1 antigenic variants were found to be circulated within the South Asian region. This emphasizes a possibility of formulating synthetic vaccines for controlling foot-and-mouth disease by Asia1 serotypes.Keywords: protein modeling, antigenic sites, sequence variation

Tube potential can be lowered to 80 kVp in test bolus phase of CT coronary angiography (CTCA) and CT pulmonary angiography (CTPA) to save dose without compromising diagnostic quality

International Nuclear Information System (INIS)

Rodrigues, J.C.L.; Manghat, N.E.; Hamilton, M.C.K.; Joshi, D.; Lyen, S.M.; Negus, I.S.

2014-01-01

The purpose of this study was to determine whether performing the test bolus (TB) of computed tomography coronary angiography (CTCA) and computed tomography pulmonary angiography (CTPA) at 80 kVp reduces dose without compromising diagnostic quality. An 80 kVp TB protocol for CTCA and CTPA was retrospectively compared to standard TB protocol (non-obese: 100 kVp, obese: 120 kVp). CT angiogram parameters were unchanged between cohorts. Thirty-seven consecutive 80 kVp TB CTCA images were compared to 53 standard CTCA images. Fifty consecutive CTPAs from each protocol were analysed. Diagnostic quality of the CT angiogram was assessed by: mean attenuation, signal-to-noise ratio (SNR) in the ascending aorta (AA) in CTCA and in the main pulmonary artery (MPA) in CTPA, diagnostic rate, and number of repeated monitoring scans. Mean effective dose was estimated using the dose-length product. Mean TB effective doses were significantly lower (P < 0.0001) for 80 kVp scans compared to the standard in non-obese CTCA (0.15 ± 0.04 mSv Vs 0.33 ± 0.09 mSv), obese CTCA (0.17 ± 0.06 mSv Vs 0.57 ± 0.12 mSv), and CTPA patients (0.07 ± 0.03 mSv Vs 0.15 ± 0.06 mSv). No difference was demonstrated in mean attenuation, SNR (AA), SNR (MPA), diagnostic rates, or number of repeated monitoring scans between protocols. Routinely performing TB at 80 kVp, regardless of body habitus, in CTCA and CTPA results in a small but significant dose reduction, without compromising CT angiogram diagnostic quality. (orig.)

Tube potential can be lowered to 80 kVp in test bolus phase of CT coronary angiography (CTCA) and CT pulmonary angiography (CTPA) to save dose without compromising diagnostic quality

Energy Technology Data Exchange (ETDEWEB)

Rodrigues, J.C.L.; Manghat, N.E.; Hamilton, M.C.K. [University Hospitals Bristol NHS Foundation Trust, Department of Radiology, Bristol Royal Infirmary, Bristol (United Kingdom); University Hospitals Bristol NHS Foundation Trust, National Institute for Health Research (NIHR) Cardiovascular Biomedical Research Unit, Bristol Heart Institute, Bristol Royal Infirmary, Bristol (United Kingdom); Joshi, D.; Lyen, S.M. [University Hospitals Bristol NHS Foundation Trust, Department of Radiology, Bristol Royal Infirmary, Bristol (United Kingdom); Negus, I.S. [University Hospitals Bristol NHS Foundation Trust, Department of Medical Physics and Bioengineering, Bristol Royal Infirmary, Bristol (United Kingdom)

2014-10-15

The purpose of this study was to determine whether performing the test bolus (TB) of computed tomography coronary angiography (CTCA) and computed tomography pulmonary angiography (CTPA) at 80 kVp reduces dose without compromising diagnostic quality. An 80 kVp TB protocol for CTCA and CTPA was retrospectively compared to standard TB protocol (non-obese: 100 kVp, obese: 120 kVp). CT angiogram parameters were unchanged between cohorts. Thirty-seven consecutive 80 kVp TB CTCA images were compared to 53 standard CTCA images. Fifty consecutive CTPAs from each protocol were analysed. Diagnostic quality of the CT angiogram was assessed by: mean attenuation, signal-to-noise ratio (SNR) in the ascending aorta (AA) in CTCA and in the main pulmonary artery (MPA) in CTPA, diagnostic rate, and number of repeated monitoring scans. Mean effective dose was estimated using the dose-length product. Mean TB effective doses were significantly lower (P < 0.0001) for 80 kVp scans compared to the standard in non-obese CTCA (0.15 ± 0.04 mSv Vs 0.33 ± 0.09 mSv), obese CTCA (0.17 ± 0.06 mSv Vs 0.57 ± 0.12 mSv), and CTPA patients (0.07 ± 0.03 mSv Vs 0.15 ± 0.06 mSv). No difference was demonstrated in mean attenuation, SNR (AA), SNR (MPA), diagnostic rates, or number of repeated monitoring scans between protocols. Routinely performing TB at 80 kVp, regardless of body habitus, in CTCA and CTPA results in a small but significant dose reduction, without compromising CT angiogram diagnostic quality. (orig.)

Expression of the multimeric and highly immunogenic Brucella spp. lumazine synthase fused to bovine rotavirus VP8d as a scaffold for antigen production in tobacco chloroplasts

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Edgardo Federico Alfano

2015-12-01

Full Text Available Lumazine synthase from Brucella spp. (BLS is a highly immunogenic decameric protein which can accommodate foreign polypeptides or protein domains fused to its N-termini, markedly increasing their immunogenicity.The inner core domain (VP8d of VP8 spike protein from bovine rotavirus (BRV is responsible for viral adhesion to sialic acid residues and infection. It also displays neutralizing epitopes, making it a good candidate for vaccination.In this work, the BLS scaffold was assessed for the first time in plants for recombinant vaccine development by N-terminally fusing BLS to VP8d and expressing the resulting fusion (BLSVP8d in tobacco chloroplasts. Transplastomic plants were obtained and characterized by Southern, northern and western blot. BLSVP8d was highly expressed, representing 40% of total soluble protein (TSP (4.85 mg/g fresh tissue. BLSVP8d remained soluble and stable during all stages of plant development and even in lyophilized leaves stored at room temperature. Soluble protein extracts from fresh and lyophilized leaves were able to induce specific neutralizing IgY antibodies in a laying hen model. This work presents BLS as an interesting platform for highly immunogenic injectable, or even oral, subunit vaccines. Lyophilization of transplastomic leaves expressing stable antigenic fusions to BLS would further reduce costs and simplify downstream processing, purification and storage, allowing for more practical vaccines.

Evaluation of a novel scoring and grading model for VP-based exams in postgraduate nurse education.

Science.gov (United States)

Forsberg, Elenita; Ziegert, Kristina; Hult, Håkan; Fors, Uno

2015-12-01

For Virtual Patient-based exams, several scoring and grading methods have been proposed, but none have yet been validated. The aim of this study was to evaluate a new scoring and grading model for VP-based exams in postgraduate paediatric nurse education. The same student group of 19 students performed a VP-based exam in three consecutive courses. When using the scoring and grading assessment model, which contains a deduction system for unnecessary or unwanted actions, a progression was found in the three courses: 53% of the students passed the first exam, 63% the second and 84% passed the final exam. The most common reason for deduction of points was due to students asking too many interview questions or ordering too many laboratory tests. The results showed that the new scoring model made it possible to judge the students' clinical reasoning process as well as their progress. Copyright © 2015 Elsevier Ltd. All rights reserved.

Direct Observation on Spin-Coating Process of PS- b -P2VP Thin Films

Energy Technology Data Exchange (ETDEWEB)

Ogawa, Hiroki; Takenaka, Mikihito; Miyazaki, Tsukasa; Fujiwara, Akihiko; Lee, Byeongdu; Shimokita, Keisuke; Nishibori, Eiji; Takata, Masaki

2016-05-10

We studied the structural development of symmetric poly(styrene-b-2-vinylpyridine) (PS-b-P2VP) block copolymers during spin-coating using in situ grazing incidence small angle X-ray scattering (GISAXS) measurements. During the spin-coating process, after the formation of the micelles in dilute solution, the selective solvent induced two kinds of the morphological transition. Firstly, the disordered spherical micelles were transformed into a BCC lattice of spheres of which the (110) plane was oriented perpendicularly to the substrate surface. Secondly, further evaporation induced a transition from spheres on the BCC lattice into cylindrical structures. The orientation of the cylinders perpendicular to the substrate surface was induced by solvent convection perpendicular to the substrate, which occurs during rapid solvent evaporation. After this transition, vitrification of PS and P2VP prevented any further transition from cylinders to the more thermodynamically stable lamellar structures, as are generally observed as the bulk equilibrium state.

Influence of minor displacements in loops of the porcine parvovirus VP2 capsid on virus-like particles assembly and the induction of antibody responses.

Science.gov (United States)

Pan, Qunxing; He, Kongwang; Wang, Yongshan; Wang, Xiaoli; Ouyang, Wei

2013-06-01

An antigen-delivery system based on hybrid virus-like particles (VLPs) formed by the self-assembly of the capsid VP2 protein of porcine parvovirus (PPV) and expressing foreign peptides offers an alternative method for vaccination. In this study, the three-dimensional structure of the PPV capsid protein and surface loops deletion mutants were analyzed to define essential domains in PPV VP2 for the assembly of VLPs. Electron microscopic analysis and SDS-PAGE analysis confirmed the presence of abundant VLPs in a loop2 deletion mutant of expected size and appropriate morphology. Loop4 and loop2-loop4 deletion mutants, however, resulted in a lower number of particles and the morphology of the particles was not well preserved. Furthermore, the green fluorescent protein (gfp) gene was used as a model. GFP was observed at the same level in displacements mutants. However, GFP displacement mutants in loop2 construct allowed better adaptation for the fusion GFP to be further displayed on the surface of the capsid-like structure. Immunogenicity study showed that there is no obvious difference in mice inoculated with rAd-VP2(Δloop2), rAd-VP2(Δloop4), rAd-VP2(Δloop2-Δloop4), and PPV inactivated vaccine. The results suggested the possibility of inserting simultaneously B and T cell epitopes in the surface loop2 and the N-terminus. The combination of different types of epitopes (B, CD4+, and CD8+) in different positions of the PPV particles opens the way to the development of highly efficient vaccines, able to stimulate at the same time the different branches of the immune system.

Analysis of the inhibitory effects of VP-16-213 (etoposide) and podophyllotoxin on thymidine transport and metabolism in Ehrlich ascites tumor cells in vitro

International Nuclear Information System (INIS)

Yalowich, J.C.; Goldman, I.D.

1984-01-01

Uptake of 3 H after exposure of cells to [ 3 H]-thymidine is characterized by a rapid initial velocity that approximates membrane transport followed by a slower rate of uptake that parallels the accumulation of phosphorylated derivatives of thymidine, primarily thymidine triphosphate, within the cell. The high rate of thymidine transport relative to thymidine metabolism to the triphosphate within the cell decreases as the extracellular nucleoside concentration is reduced due to a much greater decrease in membrane transport than the subsequent metabolic step. Hence, as extracellular thymidine is decreased, transport becomes increasingly rate limiting to metabolism within the cell. VP-16-213 (etoposide) or podophyllotoxin inhibits the initial uptake rate for thymidine and, as a consequence, inhibits the intracellular formation of thymidine triphosphate. When extracellular thymidine is high, inhibitory effects on transport are transient, and the net rate of thymidine triphosphate accumulation within drug-treated cells rapidly approaches a velocity comparable to that of control cells, indicating no direct VP-16-213 or podophyllotoxin effect on nucleoside and nucleotide phosphorylation. When extracellular thymidine is reduced so that transport is rate limiting to metabolism, the duration of the inhibitory effects of VP-16-213 on thymidine triphosphate formation is prolonged. A secondary effect of VP-16-213 becomes manifest beyond 10 min of incubation with [3H]thymidine with the virtual complete cessation of thymidine incorporation into the acid precipitate without any change in the thymidine triphosphate level. This late effect is not observed with podophyllotoxin and indicates a direct effect of VP-16-213 on DNA synthesis that is distinct from the earlier inhibitory effect on thymidine phosphorylation, which is secondary to membrane transport

A Point Mutation in the Rhesus Rotavirus VP4 Protein Generated through a Rotavirus Reverse Genetics System Attenuates Biliary Atresia in the Murine Model.

Science.gov (United States)

Mohanty, Sujit K; Donnelly, Bryan; Dupree, Phylicia; Lobeck, Inna; Mowery, Sarah; Meller, Jaroslaw; McNeal, Monica; Tiao, Greg

2017-08-01

Rotavirus infection is one of the most common causes of diarrheal illness in humans. In neonatal mice, rhesus rotavirus (RRV) can induce biliary atresia (BA), a disease resulting in inflammatory obstruction of the extrahepatic biliary tract and intrahepatic bile ducts. We previously showed that the amino acid arginine (R) within the sequence SRL (amino acids 445 to 447) in the RRV VP4 protein is required for viral binding and entry into biliary epithelial cells. To determine if this single amino acid (R) influences the pathogenicity of the virus, we generated a recombinant virus with a single amino acid mutation at this site through a reverse genetics system. We demonstrated that the RRV mutant (RRV VP4-R446G ) produced less symptomatology and replicated to lower titers both in vivo and in vitro than those seen with wild-type RRV, with reduced binding in cholangiocytes. Our results demonstrate that a single amino acid change in the RRV VP4 gene influences cholangiocyte tropism and reduces pathogenicity in mice. IMPORTANCE Rotavirus is the leading cause of diarrhea in humans. Rhesus rotavirus (RRV) can also lead to biliary atresia (a neonatal human disease) in mice. We developed a reverse genetics system to create a mutant of RRV (RRV VP4-R446G ) with a single amino acid change in the VP4 protein compared to that of wild-type RRV. In vitro , the mutant virus had reduced binding and infectivity in cholangiocytes. In vivo , it produced fewer symptoms and lower mortality in neonatal mice, resulting in an attenuated form of biliary atresia. Copyright © 2017 American Society for Microbiology.

Characteristics of a foot-and-mouth disease virus with a partial VP1 G-H loop deletion in experimentally infected cattle

DEFF Research Database (Denmark)

Fowler, Veronica; Bashiruddin, John B.; Belsham, Graham

2014-01-01

Previous work in cattle illustrated the protective efficacy and negative marker potential of a A serotype foot-and-mouth disease virus (FMDV) vaccine prepared from a virus lacking a significant portion of the VP1 G-H loop (termed A(−)). Since this deletion also includes the arginine-glycine-aspar......Previous work in cattle illustrated the protective efficacy and negative marker potential of a A serotype foot-and-mouth disease virus (FMDV) vaccine prepared from a virus lacking a significant portion of the VP1 G-H loop (termed A(−)). Since this deletion also includes the arginine...

Breast density in multiethnic women presenting for screening mammography.

Science.gov (United States)

Oppong, Bridget A; Dash, Chiranjeev; O'Neill, Suzanne; Li, Yinan; Makambi, Kepher; Pien, Edward; Makariou, Erini; Coleman, Tesha; Adams-Campbell, Lucile L

2018-05-01

Data on ethnic variations in breast density are limited and often not inclusive of underrepresented minorities. As breast density is associated with elevated breast cancer risk, investigating racial and ethnic difference may elucidate the observed differences in breast cancer risk among different populations. We reviewed breast density from initial screening of women from the Capital Breast Care Center and Georgetown University Hospital from 2010 to 2014. Patient demographics including race, age at screening, education, menopausal status, and body mass index were abstracted. We recorded the BI-RADS density categories: (1) "fatty," (2) "scattered fibroglandular densities," (3) "heterogeneously dense," and (4) "extremely dense." Multivariable unconditional logistic regression was used to identify predictors of breast density. Density categorization was recorded for 2146 women over the 5-year period, comprising Blacks (n = 940), Hispanics (n = 893), and Whites (n = 314). Analysis of subject characteristics by breast density showed that high category is observed in younger, Hispanic, nulliparous, premenopausal, and nonobese women (t-test or chi-square test, P-values density. Being Hispanic, premenopausal, and nonobese were predictive of high density on logistic regression. In this analysis of density distribution in a diverse sample, Hispanic women have the highest breast density, followed by Blacks and Whites. Unique in our findings is women who identify as Hispanic have the highest breast density and lower rates of obesity. Further investigation of the impact of obesity on breast density, especially in the understudied Hispanic group is needed. © 2017 Wiley Periodicals, Inc.

Study for evaluation of filter thickness in the determination of k Vp in function of the attenuation relative factors of the radiation

International Nuclear Information System (INIS)

Martin, J.G.; Souza, R.T.F.; Carbi, E.D.O.; Pina, D.R.

2009-01-01

This work consisted of evaluating 3 pairs of filters (cupper). The aim of it was to verify the best combination between attenuating material thicknesses for determination voltage value applied to X ray tube as function of attenuation relative factors. The employed methodology consisted of measuring the relative expositions, using thicknesses copper plates: (0,3, 0,5 and 0,8) mm. The thicknesses had been combined between itself with the purpose to form pairs of filters look that used ones ink Vp measurers. The great kVp band was used for 3 X ray equipment with generators of single-phase. Three-phase tension of 12 pulses and generator of tension in high frequency. The results had pointed the best combination, the thicknesses of filters (0,5/0,3) mm, because it does not have presented duplicity of values throughout all the band of evaluated tension. The results had still shown that the relative attenuation factors had not suffered significant variations between the different equipment with different voltage wave form. The variations found are related with differences in the effective energy of X ray beam. (author)

Verbal versus Numerical Probabilities: Does Format Presentation of Probabilistic Information regarding Breast Cancer Screening Affect Women's Comprehension?

Science.gov (United States)

Vahabi, Mandana

2010-01-01

Objective: To test whether the format in which women receive probabilistic information about breast cancer and mammography affects their comprehension. Methods: A convenience sample of 180 women received pre-assembled randomized packages containing a breast health information brochure, with probabilities presented in either verbal or numeric…

Structural basis of glycan specificity of P[19] VP8*: Implications for rotavirus zoonosis and evolution.

Science.gov (United States)

Liu, Yang; Xu, Shenyuan; Woodruff, Andrew L; Xia, Ming; Tan, Ming; Kennedy, Michael A; Jiang, Xi

2017-11-01

Recognition of specific cell surface glycans, mediated by the VP8* domain of the spike protein VP4, is the essential first step in rotavirus (RV) infection. Due to lack of direct structural information of virus-ligand interactions, the molecular basis of ligand-controlled host ranges of the major human RVs (P[8] and P[4]) in P[II] genogroup remains unknown. Here, through characterization of a minor P[II] RV (P[19]) that can infect both animals (pigs) and humans, we made an important advance to fill this knowledge gap by solving the crystal structures of the P[19] VP8* in complex with its ligands. Our data showed that P[19] RVs use a novel binding site that differs from the known ones of other genotypes/genogroups. This binding site is capable of interacting with two types of glycans, the mucin core and type 1 histo-blood group antigens (HBGAs) with a common GlcNAc as the central binding saccharide. The binding site is apparently shared by other P[II] RVs and possibly two genotypes (P[10] and P[12]) in P[I] as shown by their highly conserved GlcNAc-interacting residues. These data provide strong evidence of evolutionary connections among these human and animal RVs, pointing to a common ancestor in P[I] with a possible animal host origin. While the binding properties to GlcNAc-containing saccharides are maintained, changes in binding to additional residues, such as those in the polymorphic type 1 HBGAs may occur in the course of RV evolution, explaining the complex P[II] genogroup that mainly causes diseases in humans but also in some animals.

High prevalence of human polyomavirus JC VP1 gene sequences in pediatric malignancies.

Science.gov (United States)

Shiramizu, B; Hu, N; Frisque, R J; Nerurkar, V R

2007-05-15

The oncogenic potential of human polyomavirus JC (JCV), a ubiquitous virus that establishes infection during early childhood in approximately 70% of the human population, is unclear. As a neurotropic virus, JCV has been implicated in pediatric central nervous system tumors and has been suggested to be a pathogenic agent in pediatric acute lymphoblastic leukemia. Recent studies have demonstrated JCV gene sequences in pediatric medulloblastomas and among patients with colorectal cancer. JCV early protein T-antigen (TAg) can form complexes with cellular regulatory proteins and thus may play a role in tumorigenesis. Since JCV is detected in B-lymphocytes, a retrospective analysis of pediatric B-cell and non-B-cell malignancies as well as other HIV-associated pediatric malignancies was conducted for the presence of JCV gene sequences. DNA was extracted from 49 pediatric malignancies, including Hodgkin disease, non-Hodgkin lymphoma, large cell lymphoma and sarcoma. Polymerase chain reaction (PCR) was conducted using JCV specific nested primer sets for the transcriptional control region (TCR), TAg, and viral capsid protein 1 (VP1) genes. Southern blot analysis and DNA sequencing were used to confirm specificity of the amplicons. A 215-bp region of the JCV VP1 gene was amplified from 26 (53%) pediatric tumor tissues. The JCV TCR and two JCV gene regions were amplified from a leiomyosarcoma specimen from an HIV-infected patient. The leiomyosarcoma specimen from the cecum harbored the archetype strain of JCV. Including the leiomyosarcoma specimen, three of five specimens sequenced were typed as JCV genotype 2. The failure to amplify JCV TCR, and TAg gene sequences in the presence of JCV VP1 gene sequence is surprising. Even though JCV TAg gene, which is similar to the SV40 TAg gene, is oncogenic in animal models, the presence of JCV gene sequences in pediatric malignancies does not prove causality. In light of the available data on the presence of JCV in normal and cancerous

A CONCISE PRESENTATION OF THE FIRST LITERARY MAGANIZES PUBLISHED BY WOMEN WRITERS IN ROMANIA BETWEEN 1878 - 1947

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Carmen D. CARAIMAN

2014-05-01

Full Text Available In its early days Romanian feminine writing was influenced by the ideals of emancipation that animated the female population of our country. These ideals were shared and spread not only by feminist supporters, though this happened to a large extent, but also by outstanding men of letters (Eugen Lovinescu, Garabet-Ibraileanu and Tudor Vianu and others, who were receptive to the changing status of women in their contemporary society and encouraged it as such. The present paper focuses on what we are going to call “early feminine writing in Romania”, whereby we understand the early days of literature created by women in this country. Our goal is to identify the literary publications that gathered for the first time in Romania women writers who paid a particular interest in literature. Thus, our paper has a documentary value: it presents the main Romanian literary publications edited by the first women writers in our country, the aesthetic ideals promoted and shared by these magazines and the contribution brought by female authors to the wakening and shaping of the taste for writing and for literature amidst Romanian women in general.

Detection of uncommon G3P[3] rotavirus A (RVA) strain in rat possessing a human RVA-like VP6 and a novel NSP2 genotype.

Science.gov (United States)

Ianiro, Giovanni; Di Bartolo, Ilaria; De Sabato, Luca; Pampiglione, Guglielmo; Ruggeri, Franco M; Ostanello, Fabio

2017-09-01

Rotavirus is one of the leading causes of acute gastroenteritis in infants and young children. RVAs infect not only humans but also a wide range of mammals including rats, which represent a reservoir of several other zoonotic pathogens. Due to the segmented nature of the RVA genome, animal RVA strains can easily adapt to the human host by reassortment with co-infecting human viruses. This study aims to detect and characterize RVA in the intestinal content of Italian sinantropic rats (Rattus rattus). Out of 40 samples examined following molecular approach, one resulted positive for RVA. The molecular characterization of VP1-4, 6 and 7, and NSP1-5 genes by sequencing revealed the genomic constellation G3-P[3]-I1-R11-C11-M10-A22-N18-T14-E18-H13. This uncommon genomic combination includes: the VP1-4,VP7, the NSP1, 3, 4 and 5 gene segments, closely related to those of RVA from rodents, the N18 novel genotype established for the NSP2 gene segment and the human Wa-like VP6 gene, suggesting interspecies reassortment. Copyright © 2017 Elsevier B.V. All rights reserved.

Incidence of endometrial hyperplasia in 100 cases presenting with polymenorrhagia/menorrhagia in perimenupausal women

International Nuclear Information System (INIS)

Takreem, A.; Razaq, S.

2009-01-01

To study the Incidence of endometrial hyperplasia in perimenupausal women presenting with polymenorrhagia/menorrhagia. This observational study was conducted at Gynae 'B' unit of Khyber Teaching Hospital Peshawar from January 2000 to December 2001. One hundred consecutive patients who presented at Gynaecology OPD with Polymenorrhagia/ Menorrhagia were registered and incidence of endometrial hyperplasia evaluated in them. All women were above 45 years of age. Post-menopausal bleeding cases were excluded from the study. Out of 100 patients, 15 patients were found to have endometrial hyperplasia, 10 patients (66.6%) simple cystic hyperplasia, 3 patients (20.0%) had adenomotous hyperplasia, 2 patients (13.3%) had atypical hyperplasia, 8 patients (53.3%) with menorrhagia, 1 (6.6%) with polymenorrhagia, and 6 patients (40.0%) with polymenorrhoea. Duration of symptoms was from 4 months to 1 year. Thirteen (86.6%) patients were treated medically, 5 patients (33.33%) needed surgical treatment following medical treatment, 2 patients (13.3%) underwent Total Abdominal Hysterectomy (TAH) and Bilateral Salpingooophorectomy (BSO) who were 51-53 years of age with atypical hyperplasia. endometrial hyperplasia is a pre-malignant condition; if treated in time, incidence can be reduced and early treatment can increase life expectancy and quality in women over age of 45 years. (author)

ILC (ionic liquid colloids) based on p(4-VP) (poly(4-vinyl pyridine)) microgels: Synthesis, characterization and use in hydrogen production

International Nuclear Information System (INIS)

Sahiner, Nurettin; Turhan, Tugce; Lyon, L. Andrew

2014-01-01

In this study for the first time p(4-VP) (poly(4-vinyl pyridine)) colloidal ionic liquid particles derived from 4-VP (4-vinyl pyridine) are reported, used in the preparation of a catalyst system by loading metal salts such as CoCl 2 and NiCl 2 from ethyl alcohol solutions into the modified p(4-VP) particles, and used for hydrogen generation from NaOH-free hydrolysis of NaBH 4 . Colloidal ionic liquids containing 0.054 mmol Co and Ni were used in NaOH-free hydrolysis of 0.30 g NaBH 4 in 50 mL water at 40 °C and 1000 rpm mixing rate. The reaction rates relating to hydrolysis of NaBH 4 were 3148 (mL H 2 ) (min) −1 (g of Co) −1 for Co, and 1803 (mL H 2 ) (min) −1 (g of Ni) −1 for Ni. The effect of metal loading time, NaBH 4 concentration, temperature, and kinetic parameters were also investigated. The activation energy, enthalpy, and activation entropy for the reaction of NaBH 4 in the presence of the colloidal dicationic catalyst system were calculated as 43.98 kJ/mol, 40.38 kJ/mol, and −178.22 J/mol.K, respectively. - Highlights: • Microgel Ionic liquid colloid reactors for H 2 production. • P(4-VP) microgel ILC (ionic liquid colloid). • Modified microgel for green energy. • Ionic liquid microgel embedding metals salts NaBH 4 hydrolysis. • Ionic liquid microgel catalyst systems

The complete sequence of marine bacteriophage VpV262 infecting vibrio parahaemolyticus indicates that an ancestral component of a T7 viral supergroup is widespread in the marine environment

International Nuclear Information System (INIS)

Hardies, Stephen C.; Comeau, Andre M.; Serwer, Philip; Suttle, Curtis A.

2003-01-01

The 46,012-bp sequence of the marine bacteriophage VpV262 infecting the bacterium Vibrio parahaemolyticus is reported. The VpV262 sequence reveals that it is a distant relative of marine Roseophage SIO1, and an even more distant relative of coliphage T7. VpV262 and SIO1 appear to represent a widespread marine phage group that lacks an RNA polymerase gene and is ancestral to the T7-like phages. We propose that this group together with the T7-like phages be designated as the T7 supergroup. The ancestral head structure gene module for the T7 supergroup was reconstructed by using sensitive biased Psi-blast searches supplemented by statistical support derived from gene order. In the early and replicative segments, these phages have participated in extensive interchange with the viral gene pool. VpV262 carries a different replicative module than SIO1 and the T7-like phages

Bimolecular Complementation to Visualize Filovirus VP40-Host Complexes in Live Mammalian Cells: Toward the Identification of Budding Inhibitors

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Yuliang Liu

2011-01-01

Full Text Available Virus-host interactions play key roles in promoting efficient egress of many RNA viruses, including Ebola virus (EBOV or “e” and Marburg virus (MARV or “m”. Late- (L- domains conserved in viral matrix proteins recruit specific host proteins, such as Tsg101 and Nedd4, to facilitate the budding process. These interactions serve as attractive targets for the development of broad-spectrum budding inhibitors. A major gap still exists in our understanding of the mechanism of filovirus budding due to the difficulty in detecting virus-host complexes and mapping their trafficking patterns in the natural environment of the cell. To address this gap, we used a bimolecular complementation (BiMC approach to detect, localize, and follow the trafficking patterns of eVP40-Tsg101 complexes in live mammalian cells. In addition, we used the BiMC approach along with a VLP budding assay to test small molecule inhibitors identified by in silico screening for their ability to block eVP40 PTAP-mediated interactions with Tsg101 and subsequent budding of eVP40 VLPs. We demonstrated the potential broad spectrum activity of a lead candidate inhibitor by demonstrating its ability to block PTAP-dependent binding of HIV-1 Gag to Tsg101 and subsequent egress of HIV-1 Gag VLPs.

Nucleotide mismatches between the VP7 gene and the primer are associated with genotyping failure of a specific lineage from G1 rotavirus strains

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Espinola Emilio E

2006-05-01

Full Text Available Abstract In recent years it was reported that the accumulation of point mutations in VP4 and VP7 genes of rotavirus strains was the main cause of the failure of the G or P-typing. Failures in the correct genotyping of G1, G2, G8, G9 and G10 rotavirus strains were reported in the most commonly used reverse transcription (RT-PCR strategies. Collecting VP7 gene sequences of G1 rotavirus strains from databases we found that 74 (61.2 % out of 121 G1 strains from lineage I showed the four specific mismatches at the 5' end of the 9T1-1 primer, previously associated with the failure of G1-typing. Thus, a great percentage of the G1 strains from lineage I worldwide reported could not have been typed if the Das's RT-PCR strategy were used. This analysis shows that the failure on the detection of the G1 strains could be due to the diversification of rotavirus strains in phylogenetic lineages. Therefore, the use of different RT-PCR strategies with different primer binding locations on the VP7 gene or new typing methodologies -like microarrays procedures- could be a better option to avoid the failure of the G-typing of rotavirus strains detected during surveillance programs.

The Influence of Clinical Experience and Photographic Presentation on Age Assessment of Women

DEFF Research Database (Denmark)

Nielsen, Barbara Rubek; Linneberg, Allan; Christensen, Kaare

2016-01-01

as the presentation of participants. OBJECTIVE: It is not known whether the clinical experience of the assessor or photographic presentation have an influence on the assessment of perceived age, which the present study aimed to investigate. METHODS: In a cross-sectional study of 460 women aged 25-93 years, 10...... body. Intra- and inter-assessor agreements between photographic presentations were similar among both assessor groups. The accuracy in age assessment was significantly influenced by the photographic presentation but not by the clinical experience of the assessor. The difference in the mean perceived...... both facial and whole-body photographs. A regression towards the mean age was seen. CONCLUSION: The assessment of perceived age was influenced by the photographic presentation but not by the clinical experience of the assessor....

Non Obstetric Causes and Presentation of Acute Abdomen among the Pregnant Women.

Science.gov (United States)

Haque, Monoarul; Kamal, Farah; Chowdhury, Shahanaz; Uzzaman, Monir; Aziz, Itrat

2014-09-01

To identify the non-obstetric causes and presentation of acute abdomen among pregnant women. This was a cross sectional hospital-based study among 128 pregnant women by face to face interview using a semi-structured questionnaire. This study was conducted at the Gynecology & Obstetric Ward of 250 Bed General Hospital, Noakhali, Bangladesh, from January to August 2013. Data were analyzed by a software package used for statistical analysis (SPSS) version 11.5 (SPSS, Inc., Chicago, IL, USA). Mean age of participants was 25±4 years. Our findings showed that 81% were Muslim, 67% were lower middle income group, as well as 47% completed primary level of education. The results revealed that 28% had biliary ascariasis, 24% had peptic ulcer disease and 10% had lower urinary tract infection. We also found that 6% had acute pyelonephritis, 6% had acute gastroenteritis, 6% had acute cholecystitis, 6% had acute appendicitis, 2% had acute pancreatitis, 3% had choledocolithiasis, 2% had ovarian solid mass, 2% had twisted ovarian cyst, 4% had renal colic, and 1% had renal calculus. In non-obstetrical presentation of acute abdomen, the study found that 84% of respondents complained their pain lasting more than 24 hours. Besides, half of respondents felt pain in epigastrium and right hypochondrium. Cramping, prickling and aching type of pain were more, while 66% suffered from continuous pain. Our results also showed that 73% did not explain any aggravating factor and relieving factor, and the rest said food, fasting state and position change aggravated pain as well as relieved pain. The study concludes that precise diagnosis of the acute abdomen in pregnant women by continual updating of abdominal assessment knowledge, and clinical skills is necessary in the management of abdominal pain in obstetric settings.

Detection of Rare G3P[19] Porcine Rotavirus Strains in Chiang Mai, Thailand, Provides Evidence for Origin of the VP4 Genes of Mc323 and Mc345 Human Rotaviruses▿

Science.gov (United States)

Maneekarn, Niwat; Khamrin, Pattara; Chan-it, Wisoot; Peerakome, Supatra; Sukchai, Sujin; Pringprao, Kidsadagon; Ushijima, Hiroshi

2006-01-01

Among 175 fecal specimens collected from diarrheic piglets during a surveillance of porcine rotavirus (PoRV) strains in Chiang Mai, Thailand, 39 (22.3%) were positive for group A rotaviruses. Of these, 33.3% (13 of 39) belonged to G3P[19], which was a rare P genotype seldom reported. Interestingly, their VP4 nucleotide sequences were most closely related to human P[19] strains (Mc323 and Mc345) isolated in 1989 from the same geographical area where these PoRV strains were isolated. These P[19] PoRV strains were also closely related to another human P[19] strain (RMC321), isolated from India in 1990. The VP4 sequence identities with human P[19] were 95.4% to 97.4%, while those to a porcine P[19] strain (4F) were only 87.6 to 89.1%. Phylogenetic analysis of the VP4 gene revealed that PoRV P[19] strains clustered with human P[19] strains in a monophyletic branch separated from strain 4F. Analysis of the VP7 gene confirmed that these strains belonged to the G3 genotype and shared 97.7% to 98.3% nucleotide identities with other G3 PoRV strains circulating in the regions. This close genetic relationship was also reflected in the phylogenetic analysis of their VP7 genes. Altogether, the findings provided peculiar evidence that supported the porcine origin of VP4 genes of Mc323 and Mc345 human rotaviruses. PMID:16988014

ENDOMETRIOSIS OF APPENDIX IN WOMEN PRESENTING WITH RIGHT LOWER ABDOMINAL PAIN

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Radha Bai Prabhu T, Velayudam DA, Jayalakshmi M

2015-07-01

Full Text Available Endometriosis is a well known gynaecological condition associated with infertility and chronic pelvic pain. Review of literature shows that endometriosis can affect any tissue in the body, including the appendix. Here we report a case of pelvic endometriosis involving the vermiform appendix in a 45 years old multiparous woman. When women of the reproductive age present with recurrent lower abdominal pain on the right side, endometriosis of the appendix should also be considered. At the time of surgery appendix should be inspected and removed; especially in the presence of pelvic endometriosis.

H+ -pyrophosphatase IbVP1 promotes efficient iron use in sweet potato [Ipomoea batatas (L.) Lam.].

Science.gov (United States)

Fan, Weijuan; Wang, Hongxia; Wu, Yinliang; Yang, Nan; Yang, Jun; Zhang, Peng

2017-06-01

Iron (Fe) deficiency is one of the most common micronutrient deficiencies limiting crop production globally, especially in arid regions because of decreased availability of iron in alkaline soils. Sweet potato [Ipomoea batatas (L.) Lam.] grows well in arid regions and is tolerant to Fe deficiency. Here, we report that the transcription of type I H + -pyrophosphatase (H + -PPase) gene IbVP1 in sweet potato plants was strongly induced by Fe deficiency and auxin in hydroponics, improving Fe acquisition via increased rhizosphere acidification and auxin regulation. When overexpressed, transgenic plants show higher pyrophosphate hydrolysis and plasma membrane H + -ATPase activity compared with the wild type, leading to increased rhizosphere acidification. The IbVP1-overexpressing plants showed better growth, including enlarged root systems, under Fe-sufficient or Fe-deficient conditions. Increased ferric precipitation and ferric chelate reductase activity in the roots of transgenic lines indicate improved iron uptake, which is also confirmed by increased Fe content and up-regulation of Fe uptake genes, e.g. FRO2, IRT1 and FIT. Carbohydrate metabolism is significantly affected in the transgenic lines, showing increased sugar and starch content associated with the increased expression of AGPase and SUT1 genes and the decrease in β-amylase gene expression. Improved antioxidant capacities were also detected in the transgenic plants, which showed reduced H 2 O 2 accumulation associated with up-regulated ROS-scavenging activity. Therefore, H + -PPase plays a key role in the response to Fe deficiency by sweet potato and effectively improves the Fe acquisition by overexpressing IbVP1 in crops cultivated in micronutrient-deficient soils. © 2016 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.

K vp estimate intercomparison between Unfors XI, Radcal 4075 and a new CDTN multipurpose instrument

International Nuclear Information System (INIS)

Baptista N, A. T.; Oliveira, B. B.; Faria, L. O.

2014-08-01

This work compares results obtained using 3 (three) different instruments capable of non-invasively estimating the voltage applied to the electrodes of an x-ray emission equipment, namely the Unfors model Xi R/F, the Radcal Corporation model 4075 R/F and a new CDTN multipurpose instrument. Tests were carried out using the Pantak Seifert Model 320 Hs x-ray machine with equal setups for all instruments undergoing comparison. Irradiations were performed for different conditions of voltage and filtration. Although all instruments show a similar tendency to increase the k Vp estimate when aluminum filters are placed in the path of the x-ray beam, they may all be satisfactorily adopted in quality control routines of x-ray equipment by means of estimation of the applied voltage. The importance of using equally calibrated measurement instruments and according to manufacturers instructions became clear; in case it is not possible to follow these requirements, measurement-correcting methods must be applied. Using the new multipurpose instrument, the k Vp estimate is satisfactory even if the x-ray beam intensity is filtered in approximately one-tenth value layer. (author)

K vp estimate intercomparison between Unfors XI, Radcal 4075 and a new CDTN multipurpose instrument

Energy Technology Data Exchange (ETDEWEB)

Baptista N, A. T.; Oliveira, B. B.; Faria, L. O., E-mail: annibal@cdtn.br [Centro de Desenvolvimento da Tecnologia Nuclear - CNEN, Av. Presidente Antonio Carlos 6627, Campus UFMG, Pampulha, CEP 31270-901 Belo Horizonte, Minas Gerais (Brazil)

2014-08-15

This work compares results obtained using 3 (three) different instruments capable of non-invasively estimating the voltage applied to the electrodes of an x-ray emission equipment, namely the Unfors model Xi R/F, the Radcal Corporation model 4075 R/F and a new CDTN multipurpose instrument. Tests were carried out using the Pantak Seifert Model 320 Hs x-ray machine with equal setups for all instruments undergoing comparison. Irradiations were performed for different conditions of voltage and filtration. Although all instruments show a similar tendency to increase the k Vp estimate when aluminum filters are placed in the path of the x-ray beam, they may all be satisfactorily adopted in quality control routines of x-ray equipment by means of estimation of the applied voltage. The importance of using equally calibrated measurement instruments and according to manufacturers instructions became clear; in case it is not possible to follow these requirements, measurement-correcting methods must be applied. Using the new multipurpose instrument, the k Vp estimate is satisfactory even if the x-ray beam intensity is filtered in approximately one-tenth value layer. (author)

Men are from Mars, women are from Venus? Examining gender differences in self-presentation on social networking sites.

Science.gov (United States)

Haferkamp, Nina; Eimler, Sabrina C; Papadakis, Anna-Margarita; Kruck, Jana Vanessa

2012-02-01

Psychological research on gender differences in self-presentation has already revealed that women place higher priority on creating a positive self-presentation, while men are less concerned about the image they present in face-to-face (ftf) communication. Nowadays, with the extensive use of new media, self-presentation is no longer so closely tied to ftf situations, but can also take place in the online world. Specifically, social networking sites (SNS), such as Facebook or MySpace, offer various features such as profile pictures, groups, and virtual bulletin boards with which users can create elaborated online representations of themselves. What remains open is whether this virtual self-presentation on SNS is subject to gender differences. Based on studies emphasizing gender-related differences in Internet communication and behavior in general, it can be assumed that men and women have different motives regarding their SNS usage as well. A multimethodological study, combining results of an online survey and a content analysis of 106 user profiles, assessed users' diverse motives for participating in SNS in general, and their use of specific profile elements or self-presentation in particular. In this sample of StudiVZ users, women tend to be more likely to use SNS for comparing themselves with others and for searching for information. Men, on the other hand, are more likely to look at other people's profiles to find friends. Moreover, women tend to use group names for their self-presentation and prefer adding portrait photos to their profiles, while men choose full-body shots.

Dye-sensitized PS-b-P2VP-templated nickel oxide films for photoelectrochemical applications

OpenAIRE

Massin, Julien; Bräutigam, Maximilian; Kaeffer, Nicolas; Queyriaux, Nicolas; Field, Martin J.; Schacher, Felix H.; Popp, Jürgen; Chavarot-Kerlidou, Murielle; Dietzek, Benjamin; Artero, Vincent

2015-01-01

Moving from homogeneous water-splitting photocatalytic systems to photoelectrochemical devices requires the preparation and evaluation of novel p-type transparent conductive photoelectrode substrates. We report here on the sensitization of polystyrene-block-poly-(2-vinylpyridine) (PS-b-P2VP) diblock copolymer-templated NiO films with an organic push–pull dye. The potential of these new templated NiO film preparations for photoelectrochemical applications is compared with NiO material template...