Toxicity of selected essential oils, silicone oils, and paraffino oil against the common bed bug, cimex lectularius L. (Hemiptera: Cimicidae)

Science.gov (United States)

The common bed bug (Cimex lectularius L.) resurged in the U.S. and many other countries over the past decade. The need for safe and effective bed bug control products propelled the development of numerous “green pesticides”, mostly with essential oils listed as active ingredients. Various inorganic ...

Monitoring stink bugs (Hemiptera: Pentatomidae) in mid-Atlantic apple and peach orchards.

Science.gov (United States)

Leskey, T C; Hogmire, H W

2005-02-01

Pyramid traps coated with "industrial safety yellow" exterior latex gloss enamel paint and baited with Euschistus spp. aggregation pheromone, methyl (2E,4Z)-decadienoate captured more stink bugs than all other baited and unbaited trap types in both apple and peach orchards in 2002 and 2003. Commercial sources of dispensers of methyl (2E,4Z)-decadienoate deployed in association with pyramid traps had a significant impact on trap captures. Captures in pyramid traps were four-fold greater when baited with lures from IPM Technologies, Inc. (Portland, OR) than with lures from Suterra (Bend, OR). Variation in yellow pyramid trap color ("industrial safety yellow" and "standard coroplast yellow") and material (plywood, plastic, and masonite) did not affect trap captures. Brown stink bug was the predominant species captured (58%), followed by dusky stink bug, Euschistus tristigmus (Say) (20%); green stink bug, Acrosternum hilare (Say) (14%); and other stink bugs (Brochymena spp. and unidentified nymphs) (8%). Captures in baited pyramid traps were significantly correlated with tree beating samples in both managed and unmanaged apple orchards and with sweep netting samples in the unmanaged apple orchard. However, problems associated with trapping mechanisms of pyramid trap jar tops and jar traps likely resulted in reduced captures in baited traps. Improved trapping mechanisms must be established to develop an effective monitoring tool for stink bugs in mid-Atlantic orchards.

Distinct properties of proteases and nucleases in the gut, salivary gland and saliva of southern green stink bug, Nezara viridula

Science.gov (United States)

Lomate, Purushottam R.; Bonning, Bryony C.

2016-01-01

Stink bugs negatively impact numerous plant species of agricultural and horticultural importance. While efforts to develop effective control measures are underway, the unique digestive physiology of these pests presents a significant hurdle for either protein- or nucleotide-based management options. Here we report the comparative biochemical and proteomic characterization of proteases and nucleases from the gut, salivary gland and saliva of the southern green stink bug, Nezara viridula. The pH optimum for protease activity was acidic (5 to 6) in the gut with the primary proteases being cysteine proteases, and alkaline (8 to 9) in the saliva and salivary gland with the primary proteases being serine proteases. The serine proteases in saliva differ biochemically from trypsin and chymotrypsin, and the cathepsins in the gut and saliva showed distinct properties in inhibitor assays. Nuclease activity (DNase, RNase, dsRNase) was concentrated in the salivary gland and saliva with negligible activity in the gut. The most abundant proteins of the gut (530) and salivary gland (631) identified by proteomic analysis included four gut proteases along with eight proteases and one nuclease from the salivary gland. Understanding of N. viridula digestive physiology will facilitate the design of new strategies for management of this significant pest. PMID:27282882

Effective Bug Finding

DEFF Research Database (Denmark)

Rivas, Iago Abal

Lightweight bug finders (also known as code scanners) are becoming popular, they scale well and can find simple yet common programming errors. It is now considered a good practice to integrate these tools as part of your development process. The Linux project, for instance, has an automated testing...... service, known as the Kbuild robot, that runs a few of these code scanners. In this project, I have carefully studied tens of historical Linux bugs, and I have found that many of these bugs, despite being conceptually simple, were not caught by any code scanning tool. The reason is that, by design, code...... scanners will find mostly superficial errors. Thus, when bugs span multiple functions, even if simple, they become undetectable by most code scanners. The studied set of historical bugs contained many of such cases. This PhD thesis proposes a bug-finding technique that is both lightweight and capable...

Poliovirus-associated protein kinase: Destabilization of the virus capsid and stimulation of the phosphorylation reaction by Zn2+

International Nuclear Information System (INIS)

Ratka, M.; Lackmann, M.; Ueckermann, C.; Karlins, U.; Koch, G.

1989-01-01

The previously described poliovirus-associated protein kinase activity phosphorylates viral proteins VP0 and VP2 as well as exogenous proteins in the presence of Mg 2+ . In this paper, the effect of Zn 2+ on the phosphorylation reaction and the stability of the poliovirus capsid has been studied in detail and compared to that of Mg 2+ . In the presence of Zn 2+ , phosphorylation of capsid proteins VP2 and VP4 is significantly higher while phosphorylation of VP0 and exogenous phosphate acceptor proteins is not detected. The results indicate the activation of more than one virus-associated protein kinase by Zn 2+ . The ion-dependent behavior of the enzyme activities is observed independently of whether the virus was obtained from HeLa or green monkey kidney cells. The poliovirus capsid is destabilized by Zn 2+ . This alteration of the poliovirus capsid structure is a prerequisite for effective phosphorylation of viral capsid proteins. The increased level of phosphorylation of viral capsid proteins results in further destabilization of the viral capsid. As a result of the conformational changes, poliovirus-associated protein kinase activities dissociate from the virus particle. The authors suggest that the destabilizing effect of phosphorylation on the viral capsid plays a role in uncoating of poliovirus

R2WinBUGS: A Package for Running WinBUGS from R

Directory of Open Access Journals (Sweden)

Sibylle Sturtz

2005-01-01

Full Text Available The R2WinBUGS package provides convenient functions to call WinBUGS from R. It automatically writes the data and scripts in a format readable by WinBUGS for processing in batch mode, which is possible since version 1.4. After the WinBUGS process has finished, it is possible either to read the resulting data into R by the package itself--which gives a compact graphical summary of inference and convergence diagnostics--or to use the facilities of the coda package for further analyses of the output. Examples are given to demonstrate the usage of this package.

Structural Studies of Adeno-Associated Virus Serotype 8 Capsid Transitions Associated with Endosomal Trafficking

Energy Technology Data Exchange (ETDEWEB)

Nam, Hyun-Joo; Gurda, Brittney L.; McKenna, Robert; Potter, Mark; Byrne, Barry; Salganik, Maxim; Muzyczka, Nicholas; Agbandje-McKenna, Mavis (Florida)

2012-09-17

The single-stranded DNA (ssDNA) parvoviruses enter host cells through receptor-mediated endocytosis, and infection depends on processing in the early to late endosome as well as in the lysosome prior to nuclear entry for replication. However, the mechanisms of capsid endosomal processing, including the effects of low pH, are poorly understood. To gain insight into the structural transitions required for this essential step in infection, the crystal structures of empty and green fluorescent protein (GFP) gene-packaged adeno-associated virus serotype 8 (AAV8) have been determined at pH values of 6.0, 5.5, and 4.0 and then at pH 7.5 after incubation at pH 4.0, mimicking the conditions encountered during endocytic trafficking. While the capsid viral protein (VP) topologies of all the structures were similar, significant amino acid side chain conformational rearrangements were observed on (i) the interior surface of the capsid under the icosahedral 3-fold axis near ordered nucleic acid density that was lost concomitant with the conformational change as pH was reduced and (ii) the exterior capsid surface close to the icosahedral 2-fold depression. The 3-fold change is consistent with DNA release from an ordering interaction on the inside surface of the capsid at low pH values and suggests transitions that likely trigger the capsid for genome uncoating. The surface change results in disruption of VP-VP interface interactions and a decrease in buried surface area between VP monomers. This disruption points to capsid destabilization which may (i) release VP1 amino acids for its phospholipase A2 function for endosomal escape and nuclear localization signals for nuclear targeting and (ii) trigger genome uncoating.

Toxicities of Selected Essential Oils, Silicone Oils, and Paraffin Oil against the Common Bed Bug (Hemiptera: Cimicidae).

Science.gov (United States)

Zha, Chen; Wang, Changlu; Li, Andrew

2018-02-09

The common bed bug [Cimex lectularius L. (Hemiptera: Cimicidae)] and tropical bed bug [Cimex hemipterus F. (Hemiptera: Cimicidae)] resurged in the United States and many other countries over the past decades. The need for safe and effective bed bug control products propelled the development of numerous 'green insecticides', mostly with essential oils listed as active ingredients. Various inorganic and organic oils also were used for bed bug management. However, there are no published studies on their toxicities against bed bugs. In this study, we screened 18 essential oils, three silicone oils, and paraffin oil (C5-20 paraffins) for their toxicities against bed bugs. All the oils exhibited insecticidal activity in topical assays. Their toxicities varied significantly; all of the evaluated essential oils were less effective than silicone oils and paraffin oil. The LD50 values of the most effective essential oil (blood orange), paraffin oil, and the most effective silicone oil (dodecamethylpentasiloxane) are 0.184 ± 0.018, 0.069 ± 0.012, and 0.036 ± 0.005 mg per bug, respectively. Direct spray of 1% water solution of 3-[hydroxy (polyethyleneoxy) propyl] heptamethyltrisiloxane, the only silicone oil that mixes well with water, resulted in 92% bed bug mortality after 1 d. Results of this study indicate silicone oils and paraffin oil have the potential to be used as safer alternative bed bug control materials. © The Author(s) 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Residues of the UL25 Protein of Herpes Simplex Virus That Are Required for Its Stable Interaction with Capsids ▿

Science.gov (United States)

Cockrell, Shelley K.; Huffman, Jamie B.; Toropova, Katerina; Conway, James F.; Homa, Fred L.

2011-01-01

The herpes simplex virus 1 (HSV-1) UL25 gene product is a minor capsid component that is required for encapsidation, but not cleavage, of replicated viral DNA. UL25 is located on the capsid surface in a proposed heterodimer with UL17, where five copies of the heterodimer are found at each of the capsid vertices. Previously, we demonstrated that amino acids 1 to 50 of UL25 are essential for its stable interaction with capsids. To further define the UL25 capsid binding domain, we generated recombinant viruses with either small truncations or amino acid substitutions in the UL25 N terminus. Studies of these mutants demonstrated that there are two important regions within the capsid binding domain. The first 27 amino acids are essential for capsid binding of UL25, while residues 26 to 39, which are highly conserved in the UL25 homologues of other alphaherpesviruses, were found to be critical for stable capsid binding. Cryo-electron microscopy reconstructions of capsids containing either a small tag on the N terminus of UL25 or the green fluorescent protein (GFP) fused between amino acids 50 and 51 of UL25 demonstrate that residues 1 to 27 of UL25 contact the hexon adjacent to the penton. A second region, most likely centered on amino acids 26 to 39, contacts the triplex that is one removed from the penton. Importantly, both of these UL25 capsid binding regions are essential for the stable packaging of full-length viral genomes. PMID:21411517

Herpesvirus capsid assembly and DNA packaging

Science.gov (United States)

Heming, Jason D.; Conway, James F.; Homa, Fred L.

2017-01-01

Herpes simplex virus type I (HSV-1) is the causative agent of several pathologies ranging in severity from the common cold sore to life-threatening encephalitic infection. During productive lytic infection, over 80 viral proteins are expressed in a highly regulated manner, resulting in the replication of viral genomes and assembly of progeny virions. The virion of all herpesviruses consists of an external membrane envelope, a proteinaceous layer called the tegument, and an icosahedral capsid containing the double-stranded linear DNA genome. The capsid shell of HSV-1 is built from four structural proteins: a major capsid protein, VP5, which forms the capsomers (hexons and pentons), the triplex consisting of VP19C and VP23 found between the capsomers, and VP26 which binds to VP5 on hexons but not pentons. In addition, the dodecameric pUL6 portal complex occupies one of the 12 capsid vertices, and the capsid vertex specific component (CVSC), a heterotrimer complex of pUL17, pUL25 and pUL36 binds specifically to the triplexes adjacent to each penton. The capsid is assembled in the nucleus where the viral genome is packaged into newly assembled closed capsid shells. Cleavage and packaging of replicated, concatemeric viral DNA requires the seven viral proteins encoded by the UL6, UL15, UL17, UL25, UL28, UL32, and UL33 genes. Considerable advances have been made in understanding the structure of the herpesvirus capsid and the function of several of the DNA packaging proteins by applying biochemical, genetic, and structural techniques. This review is a summary of recent advances with respect to the structure of the HSV-1 virion capsid and what is known about the function of the seven packaging proteins and their interactions with each other and with the capsid shell. PMID:28528442

Stabilising the Herpes Simplex Virus capsid by DNA packaging

Science.gov (United States)

Wuite, Gijs; Radtke, Kerstin; Sodeik, Beate; Roos, Wouter

2009-03-01

Three different types of Herpes Simplex Virus type 1 (HSV-1) nuclear capsids can be distinguished, A, B and C capsids. These capsids types are, respectively, empty, contain scaffold proteins, or hold DNA. We investigate the physical properties of these three capsids by combining biochemical and nanoindentation techniques. Atomic Force Microscopy (AFM) experiments show that A and C capsids are mechanically indistinguishable whereas B capsids already break at much lower forces. By extracting the pentamers with 2.0 M GuHCl or 6.0 M Urea we demonstrate an increased flexibility of all three capsid types. Remarkably, the breaking force of the B capsids without pentamers does not change, while the modified A and C capsids show a large drop in their breaking force to approximately the value of the B capsids. This result indicates that upon DNA packaging a structural change at or near the pentamers occurs which mechanically reinforces the capsids structure. The reported binding of proteins UL17/UL25 to the pentamers of the A and C capsids seems the most likely candidate for such capsids strengthening. Finally, the data supports the view that initiation of DNA packaging triggers the maturation of HSV-1 capsids.

Bed Bug Infestations and Control Practices in China: Implications for Fighting the Global Bed Bug Resurgence

Directory of Open Access Journals (Sweden)

Changlu Wang

2011-04-01

Full Text Available The bed bug resurgence in North America, Europe, and Australia has elicited interest in investigating the causes of the widespread and increasing infestations and in developing more effective control strategies. In order to extend global perspectives on bed bug management, we reviewed bed bug literature in China by searching five Chinese language electronic databases. We also conducted telephone interviews of 68 pest control firms in two cities during March 2011. In addition, we conducted telephone interviews to 68 pest control companies within two cities in March 2011. Two species of bed bugs (Cimex lectularius L. and Cimex hemipterus (F. are known to occur in China. These were common urban pests before the early1980s. Nationwide “Four-Pest Elimination” campaigns (bed bugs being one of the targeted pests were implemented in China from 1960 to the early 1980s. These campaigns succeeded in the elimination of bed bug infestations in most communities. Commonly used bed bug control methods included applications of hot water, sealing of bed bug harborages, physical removal, and applications of residual insecticides (mainly organophosphate sprays or dusts. Although international and domestic travel has increased rapidly in China over the past decade (2000–2010, there have only been sporadic new infestations reported in recent years. During 1999–2009, all documented bed bug infestations were found in group living facilities (military dormitories, worker dormitories, and prisons, hotels, or trains. One city (Shenzhen city near Hong Kong experienced significantly higher number of bed bug infestations. This city is characterized by a high concentration of migratory factory workers. Current bed bug control practices include educating residents, washing, reducing clutter, putting items under the hot sun in summer, and applying insecticides (pyrethroids or organophosphates. There have not been any studies or reports on bed bug insecticide

Labeling and localization of the herpes simplex virus capsid protein UL25 and its interaction with the two triplexes closest to the penton

Science.gov (United States)

Conway, James F.; Cockrell, Shelley K.; Copeland, Anna Maria; Newcomb, William W.; Brown, Jay C.; Homa, Fred L.

2010-01-01

The herpes simplex virus type 1 (HSV-1) UL25 protein is one of seven viral proteins that are required for DNA cleavage and packaging. Together with UL17, UL25 forms part of an elongated molecule referred to as the C-capsid-specific component or CCSC. Five copies of the CCSC are located at each of the capsid vertices on DNA-containing capsids. To study the conformation of UL25 as it is folded on the capsid surface, we identified the sequence recognized by a UL25-specific monoclonal antibody and localized the epitope on the capsid surface by immunogold electron microscopy. The epitope mapped to amino acids 99-111 adjacent to the region of the protein (amino acids 1-50) that is required for capsid binding. In addition, cryo-EM reconstructions of C-capsids in which the green fluorescent protein (GFP) was fused within the N-terminus of UL25 localized the point of contact between UL25 and GFP. The result confirmed the modeled location of the UL25 protein in the CCSC density as the region that is distal to the penton with the N-terminus of UL25 making contact with the triplex one removed from the penton. Immunofluorescence experiments at early times during infection demonstrated that UL25-GFP was present on capsids located within the cytoplasm and adjacent to the nucleus. These results support the view that UL25 is present on incoming capsids with the capsid binding domain of UL25 located on the surface of the mature DNA-containing capsid. PMID:20109467

Using Software Dependency to Bug Prediction

Directory of Open Access Journals (Sweden)

Peng He

2013-01-01

Full Text Available Software maintenance, especially bug prediction, plays an important role in evaluating software quality and balancing development costs. This study attempts to use several quantitative network metrics to explore their relationships with bug prediction in terms of software dependency. Our work consists of four main steps. First, we constructed software dependency networks regarding five dependency scenes at the class-level granularity. Second, we used a set of nine representative and commonly used metrics—namely, centrality, degree, PageRank, and HITS, as well as modularity—to quantify the importance of each class. Third, we identified how these metrics were related to the proneness and severity of fixed bugs in Tomcat and Ant and determined the extent to which they were related. Finally, the significant metrics were considered as predictors for bug proneness and severity. The result suggests that there is a statistically significant relationship between class’s importance and bug prediction. Furthermore, betweenness centrality and out-degree metric yield an impressive accuracy for bug prediction and test prioritization. The best accuracy of our prediction for bug proneness and bug severity is up to 54.7% and 66.7% (top 50, Tomcat and 63.8% and 48.7% (top 100, Ant, respectively, within these two cases.

The tripartite capsid gene of Salmonella phage Gifsy-2 yields a capsid assembly pathway engaging features from HK97 and λ

International Nuclear Information System (INIS)

Effantin, Gregory; Figueroa-Bossi, Nara; Schoehn, Guy; Bossi, Lionello; Conway, James F.

2010-01-01

Phage Gifsy-2, a lambdoid phage infecting Salmonella, has an unusually large composite gene coding for its major capsid protein (mcp) at the C-terminal end, a ClpP-like protease at the N-terminus, and a ∼ 200 residue central domain of unknown function but which may have a scaffolding role. This combination of functions on a single coding region is more extensive than those observed in other phages such as HK97 (scaffold-capsid fusion) and λ (protease-scaffold fusion). To study the structural phenotype of the unique Gifsy-2 capsid gene, we have purified Gifsy-2 particles and visualized capsids and procapsids by cryoelectron microscopy, determining structures to resolutions up to 12 A. The capsids have lambdoid T = 7 geometry and are well modeled with the atomic structures of HK97 mcp and phage λ gpD decoration protein. Thus, the unique Gifsy-2 capsid protein gene yields a capsid maturation pathway engaging features from both phages HK97 and λ.

Bayesian modeling using WinBUGS

CERN Document Server

Ntzoufras, Ioannis

2009-01-01

A hands-on introduction to the principles of Bayesian modeling using WinBUGS Bayesian Modeling Using WinBUGS provides an easily accessible introduction to the use of WinBUGS programming techniques in a variety of Bayesian modeling settings. The author provides an accessible treatment of the topic, offering readers a smooth introduction to the principles of Bayesian modeling with detailed guidance on the practical implementation of key principles. The book begins with a basic introduction to Bayesian inference and the WinBUGS software and goes on to cover key topics, including: Markov Chain Monte Carlo algorithms in Bayesian inference Generalized linear models Bayesian hierarchical models Predictive distribution and model checking Bayesian model and variable evaluation Computational notes and screen captures illustrate the use of both WinBUGS as well as R software to apply the discussed techniques. Exercises at the end of each chapter allow readers to test their understanding of the presented concepts and all ...

42 Variability Bugs in the Linux Kernel

DEFF Research Database (Denmark)

Abal, Iago; Brabrand, Claus; Wasowski, Andrzej

2014-01-01

Feature-sensitive verification pursues effective analysis of the exponentially many variants of a program family. However, researchers lack examples of concrete bugs induced by variability, occurring in real large-scale systems. Such a collection of bugs is a requirement for goal-oriented research...... provide self-contained simplified C99 versions of the bugs, facilitating understanding and tool evaluation. Our study provides insights into the nature and occurrence of variability bugs in a large C software system, and shows in what ways variability affects and increases the complexity of software bugs....

Structural basis for the development of avian virus capsids that display influenza virus proteins and induce protective immunity.

Science.gov (United States)

Pascual, Elena; Mata, Carlos P; Gómez-Blanco, Josué; Moreno, Noelia; Bárcena, Juan; Blanco, Esther; Rodríguez-Frandsen, Ariel; Nieto, Amelia; Carrascosa, José L; Castón, José R

2015-03-01

Bioengineering of viruses and virus-like particles (VLPs) is a well-established approach in the development of new and improved vaccines against viral and bacterial pathogens. We report here that the capsid of a major avian pathogen, infectious bursal disease virus (IBDV), can accommodate heterologous proteins to induce protective immunity. The structural units of the ~70-nm-diameter T=13 IBDV capsid are trimers of VP2, which is made as a precursor (pVP2). The pVP2 C-terminal domain has an amphipathic α helix that controls VP2 polymorphism. In the absence of the VP3 scaffolding protein, 466-residue pVP2 intermediates bearing this α helix assemble into genuine VLPs only when expressed with an N-terminal His6 tag (the HT-VP2-466 protein). HT-VP2-466 capsids are optimal for protein insertion, as they are large enough (cargo space, ~78,000 nm(3)) and are assembled from a single protein. We explored HT-VP2-466-based chimeric capsids initially using enhanced green fluorescent protein (EGFP). The VLP assembly yield was efficient when we coexpressed EGFP-HT-VP2-466 and HT-VP2-466 from two recombinant baculoviruses. The native EGFP structure (~240 copies/virion) was successfully inserted in a functional form, as VLPs were fluorescent, and three-dimensional cryo-electron microscopy showed that the EGFP molecules incorporated at the inner capsid surface. Immunization of mice with purified EGFP-VLPs elicited anti-EGFP antibodies. We also inserted hemagglutinin (HA) and matrix (M2) protein epitopes derived from the mouse-adapted A/PR/8/34 influenza virus and engineered several HA- and M2-derived chimeric capsids. Mice immunized with VLPs containing the HA stalk, an M2 fragment, or both antigens developed full protection against viral challenge. Virus-like particles (VLPs) are multimeric protein cages that mimic the infectious virus capsid and are potential candidates as nonliving vaccines that induce long-lasting protection. Chimeric VLPs can display or include foreign

Determination of prestress and elastic properties of virus capsids

Science.gov (United States)

Aggarwal, Ankush

2018-03-01

Virus capsids are protein shells that protect the virus genome, and determination of their mechanical properties has been a topic of interest because of their potential use in nanotechnology and therapeutics. It has been demonstrated that stresses exist in virus capsids, even in their equilibrium state, due to their construction. These stresses, termed "prestresses" in this study, closely affect the capsid's mechanical behavior. Three methods—shape-based metric, atomic force microscope indentation, and molecular dynamics—have been proposed to determine the capsid elastic properties without fully accounting for prestresses. In this paper, we theoretically analyze the three methods used for mechanical characterization of virus capsids and numerically investigate how prestresses affect the capsid's mechanical properties. We consolidate all the results and propose that by using these techniques collectively, it is possible to accurately determine both the mechanical properties and prestresses in capsids.

Bug City: Beetles [Videotape].

Science.gov (United States)

1998

"Bug City" is a video series created to help children learn about insects and other small critters. All aspects of bug life are touched upon including body structure, food, habitat, life cycle, mating habits, camouflage, mutualism (symbiosis), adaptations, social behavior, and more. Each program features dramatic microscopic photography,…

Bug City: Bees [Videotape].

Science.gov (United States)

1998

"Bug City" is a video series created to help children learn about insects and other small critters. All aspects of bug life are touched upon including body structure, food, habitat, life cycle, mating habits, camouflage, mutualism (symbiosis), adaptations, social behavior, and more. Each program features dramatic microscopic photography,…

Bug City: Ants [Videotape].

Science.gov (United States)

1998

"Bug City" is a video series created to help children (grades 1-6) learn about insects and other small critters. All aspects of bug life are touched upon including body structure, food, habitat, life cycle, mating habits, camouflage, mutualism (symbiosis), adaptations, social behavior, and more. Each program features dramatic microscopic…

40 Variability Bugs in the Linux Kernel

DEFF Research Database (Denmark)

Abal Rivas, Iago; Brabrand, Claus; Wasowski, Andrzej

2014-01-01

Feature-sensitive verification is a recent field that pursues the effective analysis of the exponential number of variants of a program family. Today researchers lack examples of concrete bugs induced by variability, and occurring in real large-scale software. Such a collection of bugs is a requi......Feature-sensitive verification is a recent field that pursues the effective analysis of the exponential number of variants of a program family. Today researchers lack examples of concrete bugs induced by variability, and occurring in real large-scale software. Such a collection of bugs...... the outcome of our analysis into a database. In addition, we provide self-contained simplified C99 versions of the bugs, facilitating understanding and tool evaluation. Our study provides insights about the nature and occurrence of variability bugs in a large C software system, and shows in what ways...

Stochastic modeling of virus capsid assembly pathways

Science.gov (United States)

Schwartz, Russell

2009-03-01

Virus capsids have become a key model system for understanding self-assembly due to their high complexity, robust and efficient assembly processes, and experimental tractability. Our ability to directly examine and manipulate capsid assembly kinetics in detail nonetheless remains limited, creating a need for computer models that can infer experimentally inaccessible features of the assembly process and explore the effects of hypothetical manipulations on assembly trajectories. We have developed novel algorithms for stochastic simulation of capsid assembly [1,2] that allow us to model capsid assembly over broad parameter spaces [3]. We apply these methods to study the nature of assembly pathway control in virus capsids as well as their sensitivity to assembly conditions and possible experimental interventions. [4pt] [1] F. Jamalyaria, R. Rohlfs, and R. Schwartz. J Comp Phys 204, 100 (2005). [0pt] [2] N. Misra and R. Schwartz. J Chem Phys 129, in press (2008). [0pt] [3] B. Sweeney, T. Zhang, and R. Schwartz. Biophys J 94, 772 (2008).

Polarized DNA Ejection from the Herpesvirus Capsid

Science.gov (United States)

Newcomb, William W.; Cockrell, Shelley K.; Homa, Fred L.; Brown, Jay C.

2009-01-01

Ejection of DNA from the capsid is an early step in infection by all herpesviruses. Ejection or DNA uncoating occurs after a parental capsid has entered the host cell cytoplasm, migrated to the nucleus and bound to a nuclear pore. DNA exits the capsid through the portal vertex and proceeds by way of the nuclear pore complex into the nucleoplasm where it is transcribed and replicated. Here we describe use of an in vitro uncoating system to determine which genome end exits first from the herpes simplex virus (HSV-1) capsid. Purified DNA-containing capsids were bound to a solid surface and warmed under conditions in which some, but not all, of the DNA was ejected. Restriction endonuclease digestion was then used to identify the genomic origin of the ejected DNA. The results support the view that the S segment end exits the capsid first. Preferential release at the S end demonstrates that herpesvirus DNA uncoating conforms to the paradigm in dsDNA bacteriophage where the last end packaged is the first to be ejected. Release of HSV-1 DNA beginning at the S end causes the first gene to enter the host cell nucleus to be α4, a transcription factor required for expression of early genes. PMID:19631662

Large-scale functional purification of recombinant HIV-1 capsid.

Directory of Open Access Journals (Sweden)

Magdeleine Hung

Full Text Available During human immunodeficiency virus type-1 (HIV-1 virion maturation, capsid proteins undergo a major rearrangement to form a conical core that protects the viral nucleoprotein complexes. Mutations in the capsid sequence that alter the stability of the capsid core are deleterious to viral infectivity and replication. Recently, capsid assembly has become an attractive target for the development of a new generation of anti-retroviral agents. Drug screening efforts and subsequent structural and mechanistic studies require gram quantities of active, homogeneous and pure protein. Conventional means of laboratory purification of Escherichia coli expressed recombinant capsid protein rely on column chromatography steps that are not amenable to large-scale production. Here we present a function-based purification of wild-type and quadruple mutant capsid proteins, which relies on the inherent propensity of capsid protein to polymerize and depolymerize. This method does not require the packing of sizable chromatography columns and can generate double-digit gram quantities of functionally and biochemically well-behaved proteins with greater than 98% purity. We have used the purified capsid protein to characterize two known assembly inhibitors in our in-house developed polymerization assay and to measure their binding affinities. Our capsid purification procedure provides a robust method for purifying large quantities of a key protein in the HIV-1 life cycle, facilitating identification of the next generation anti-HIV agents.

Bovine adenovirus type 3 containing heterologous protein in the C-terminus of minor capsid protein IX

International Nuclear Information System (INIS)

Zakhartchouk, Alexander; Connors, Wayne; Van Kessel, Andrew; Tikoo, Suresh Kumar

2004-01-01

Earlier, we detected pIX of BAdV-3 as a 14-kDa protein in purified virions. Analysis of BAdV-3 pIX using different region antibodies revealed that the N-terminus and central domain of the pIX contain immunogenic sites and are not exposed on the surface of BAdV-3 virion. This suggested that the C-terminus of BAdV-3 pIX (125 amino acid) may be exposed on the virion and may be used as a site for incorporation of heterologous peptides or proteins. We constructed recombinant BAV950 containing a small peptide (21 amino acid), including the RGD motif or recombinant BAV951 containing enhanced yellow-green fluorescent protein (EYFP) fused to the C-terminus of pIX. Western blot analysis demonstrated that the chimeric pIX-RGD was incorporated into virion capsids. Incorporation of the RGD motif into the pIX resulted in significant augmentation of BAdV-3 fiber knob-independent infection of the integrin-positive cells, suggesting that RGD motifs are displayed on the surface of virion capsids and are accessible for binding to integrins. Analysis of BAV951 revealed that the chimeric pIX is incorporated into virion capsids and EYFP containing the C-terminus of pIX is exposed on the surface of the virion. Moreover, insertion of chimeric pIXs was maintained without change through successive rounds of viral replication. These results suggested that in contrast to major capsid proteins (hexon, penton, fiber), the minor capsid protein IX can be use for the incorporation of targeting ligands based on either small peptides or longer polypeptides

Biophysical characterization of the feline immunodeficiency virus p24 capsid protein conformation and in vitro capsid assembly.

Directory of Open Access Journals (Sweden)

Jennifer Serrière

Full Text Available The Feline Immunodeficiency Virus (FIV capsid protein p24 oligomerizes to form a closed capsid that protects the viral genome. Because of its crucial role in the virion, FIV p24 is an interesting target for the development of therapeutic strategies, although little is known about its structure and assembly. We defined and optimized a protocol to overexpress recombinant FIV capsid protein in a bacterial system. Circular dichroism and isothermal titration calorimetry experiments showed that the structure of the purified FIV p24 protein was comprised mainly of α-helices. Dynamic light scattering (DLS and cross-linking experiments demonstrated that p24 was monomeric at low concentration and dimeric at high concentration. We developed a protocol for the in vitro assembly of the FIV capsid. As with HIV, an increased ionic strength resulted in FIV p24 assembly in vitro. Assembly appeared to be dependent on temperature, salt concentration, and protein concentration. The FIV p24 assembly kinetics was monitored by DLS. A limit end-point diameter suggested assembly into objects of definite shapes. This was confirmed by electron microscopy, where FIV p24 assembled into spherical particles. Comparison of FIV p24 with other retroviral capsid proteins showed that FIV assembly is particular and requires further specific study.

Bug bites and stings: When to see a dermatologist

Medline Plus

Full Text Available ... Tips to prevent and treat bug bites Although most bug bites are harmless, some can spread dangerous ... one’s greatest efforts, bug bites still happen. Fortunately, most bug bites and stings can be safely treated ...

Effectiveness of Bed Bug Pesticides

Science.gov (United States)

Before EPA allows a bed bug claim on a label, the product must be supported by data showing it will kill bed bugs when applied according to the label. Also consider factors such as extent of infestation, site preparation, and insect life stages.

Periodic table of virus capsids: implications for natural selection and design.

Science.gov (United States)

Mannige, Ranjan V; Brooks, Charles L

2010-03-04

For survival, most natural viruses depend upon the existence of spherical capsids: protective shells of various sizes composed of protein subunits. So far, general evolutionary pressures shaping capsid design have remained elusive, even though an understanding of such properties may help in rationally impeding the virus life cycle and designing efficient nano-assemblies. This report uncovers an unprecedented and species-independent evolutionary pressure on virus capsids, based on the the notion that the simplest capsid designs (or those capsids with the lowest "hexamer complexity", C(h)) are the fittest, which was shown to be true for all available virus capsids. The theories result in a physically meaningful periodic table of virus capsids that uncovers strong and overarching evolutionary pressures, while also offering geometric explanations to other capsid properties (rigidity, pleomorphy, auxiliary requirements, etc.) that were previously considered to be unrelatable properties of the individual virus. Apart from describing a universal rule for virus capsid evolution, our work (especially the periodic table) provides a language with which highly diverse virus capsids, unified only by geometry, may be described and related to each other. Finally, the available virus structure databases and other published data reiterate the predicted geometry-derived rules, reinforcing the role of geometry in the natural selection and design of virus capsids.

An elastic network model of HK97 capsid maturation.

Science.gov (United States)

Kim, Moon K; Jernigan, Robert L; Chirikjian, Gregory S

2003-08-01

The structure of the capsid of bacteriophage HK97 has been solved at various stages of maturity by crystallography and cryo-electron microscopy, and has been reported previously in the literature. Typically the capsid assembles through polymerization and maturation processes. Maturation is composed of proteolytic cleavages to the precursor capsid (called Prohead II), expansion triggered by DNA packaging (in which the largest conformational changes of the capsid appear), and covalent cross-links of neighboring subunits to create the mature capsid called Head II. We apply a coarse-grained elastic network interpolation (ENI) to generate a feasible pathway for conformational change from Prohead II to Head II. The icosahedral symmetry of the capsid structure offers a significant computational advantage because it is not necessary to consider the whole capsid structure but only an asymmetric unit consisting of one hexamer plus an additional subunit from an adjacent pentamer. We also analyze normal modes of the capsid structure using an elastic network model which is also subject to symmetry constraints. Using our model, we can visualize the smooth evolution of capsid expansion and revisit in more detail several interesting geometric changes recognized in early experimental works such as rigid body motion of two compact domains (A and P) with two refolding extensions (N-arm and E-loop) and track the approach of the two particular residues associated with isopeptide bonds that make hexagonal cross-links in Head II. The feasibility of the predicted pathway is also supported by the results of our normal mode analysis.

Bug22p, a conserved centrosomal/ciliary protein also present in higher plants, is required for an effective ciliary stroke in Paramecium.

Science.gov (United States)

Laligné, C; Klotz, C; de Loubresse, N Garreau; Lemullois, M; Hori, M; Laurent, F X; Papon, J F; Louis, B; Cohen, J; Koll, F

2010-04-01

Centrioles, cilia, and flagella are ancestral conserved organelles of eukaryotic cells. Among the proteins identified in the proteomics of ciliary proteins in Paramecium, we focus here on a protein, Bug22p, previously detected by cilia and basal-body high-throughput studies but never analyzed per se. Remarkably, this protein is also present in plants, which lack centrioles and cilia. Bug22p sequence alignments revealed consensus positions that distinguish species with centrioles/cilia from plants. In Paramecium, antibody and green fluorescent protein (GFP) fusion labeling localized Bug22p in basal bodies and cilia, and electron microscopy immunolabeling refined the localization to the terminal plate of the basal bodies, the transition zone, and spots along the axoneme, preferentially between the membrane and the microtubules. RNA interference (RNAi) depletion of Bug22p provoked a strong decrease in swimming speed, followed by cell death after a few days. High-speed video microscopy and morphological analysis of Bug22p-depleted cells showed that the protein plays an important role in the efficiency of ciliary movement by participating in the stroke shape and rigidity of cilia. The defects in cell swimming and growth provoked by RNAi can be complemented by expression of human Bug22p. This is the first reported case of complementation by a human gene in a ciliate.

The BUG BITBUS Universal Gateway

International Nuclear Information System (INIS)

Nawrocki, G.

1996-01-01

The BITBUS Universal Gateway (BUG) provides a unique, cost effective solution to many different computer interface problems. Each BUG node, utilizing ''on board'' intelligence, has the ability to provide a communication link between BITBUS protocol and other computer-signal interfaces. Among them, IEEE-488, RS232, and raw analog and binary signal 1/0. BITBUS is a multidrop, multinode link with the ability to communicate over great distances. By using this method of signal transfer, along with the communication conversion ability of the BUG, one could accomplish such things as the ability to run IEEE-488 instruments over great distances, extend a multidrop links to RS232 instruments, and provide a convenient interface point for remote analog and binary 1/0 signals, all on one homogeneous network. The BUG not only provides this through the wired ''twisted pair'' standard of BITBUS, but extends the ability to fiber optic communications for signal transfer over extreme distances and through electrically ''noisy'' environments

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

Science.gov (United States)

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

Properties and Functions of the Dengue Virus Capsid Protein.

Science.gov (United States)

Byk, Laura A; Gamarnik, Andrea V

2016-09-29

Dengue virus affects hundreds of millions of people each year around the world, causing a tremendous social and economic impact on affected countries. The aim of this review is to summarize our current knowledge of the functions, structure, and interactions of the viral capsid protein. The primary role of capsid is to package the viral genome. There are two processes linked to this function: the recruitment of the viral RNA during assembly and the release of the genome during infection. Although particle assembly takes place on endoplasmic reticulum membranes, capsid localizes in nucleoli and lipid droplets. Why capsid accumulates in these locations during infection remains unknown. In this review, we describe available data and discuss new ideas on dengue virus capsid functions and interactions. We believe that a deeper understanding of how the capsid protein works during infection will create opportunities for novel antiviral strategies, which are urgently needed to control dengue virus infections.

Find a Bed Bug Pesticide Product

Science.gov (United States)

Introduces the Bed Bug Product Search Tool, to help consumers find EPA-registered pesticides for bed bug infestation control. Inclusion in this database is not an endorsement. Always follow label directions carefully.

Bugs, bees and spiders : green roof design for rare invertebrates

Energy Technology Data Exchange (ETDEWEB)

Gedge, D. [Livingroofs.org, London (United Kingdom); Kadas, G. [Royal Holloway Univ. of London, London (United Kingdom)

2004-07-01

The use of green roofs as mitigation technique for biodiversity is particularly relevant for the objectives of the London Biodiversity Partnership, particularly since London is undergoing large-scale regeneration and many of the new developments will be targeted on brownfield land. In 2002 two research projects were undertaken to create a baseline of data on how invertebrates were using the current green roofs in London. The London Biodiversity Partnership's Black Redstart Action Plan conducts research into green roofs to demonstrate how they can be maximized for biodiversity. The Black Redstart Project ensures that green roofs are used in new developments in London where such developments threaten this species. It is one of the country's rarest breeding birds, and is unique in that it is predominantly found in cities, on brownfield sites and post-industrial sites. Three green roof laboratories were established at 2 sites in London to investigate how substrates, substrate depths and planting affects the fauna associated with brownfields and green roofs in London. Although conservationists in London have urged many developers to provide green roofs to help the Black Redstart, there is concern that many of these roofs do no provide the proper support for the species. In some cases roofs are constructed of commercially driven products such as sedum mats that do provide habitat for some rare invertebrates but are not as supportive of a greater diversity of species as they could be due to the design process and a lack of knowledge of green roof technology. It was suggested that there is a need for cooperation between ecologists and Architects in order to achieve the habitat. 18 refs., 6 figs.

Bug City: Aquatic Insects [Videotape].

Science.gov (United States)

1998

"Bug City" is a video series created to help children learn about insects and other small critters. All aspects of bug life are touched upon including body structure, food, habitat, life cycle, mating habits, camouflage, mutualism (symbiosis), adaptations, social behavior, and more. Each program features dramatic microscopic photography,…

Bug City: Flies & Mosquitoes [Videotape].

Science.gov (United States)

1998

"Bug City" is a video series created to help children (grades 1-6) learn about insects and other small critters. All aspects of bug life are touched upon, including body structure, food, habitat, life cycle, mating habits, camouflage, mutualism (symbiosis), adaptations, social behavior, and more. Each program features dramatic…

Bug City: Ladybugs & Fireflies [Videotape].

Science.gov (United States)

1998

"Bug City" is a video series created to help children (grades 1-6) learn about insects and other small critters. All aspects of bug life are touched upon, including body structure, food, habitat, life cycle, mating habits, camouflage, mutualism (symbiosis), adaptations, social behavior, and more. Each program features dramatic…

Towards Easing the Diagnosis of Bugs in OS Code

DEFF Research Database (Denmark)

Stuart, Henrik; Hansen, René Rydhof; Lawall, Julia Laetitia

2007-01-01

The rapid detection and treatment of bugs in operating systems code is essential to maintain the overall security and dependability of a computing system.  A number of techniques have been proposed for detecting bugs, but little has been done to help developers analyze and treat them.  In this pa......The rapid detection and treatment of bugs in operating systems code is essential to maintain the overall security and dependability of a computing system.  A number of techniques have been proposed for detecting bugs, but little has been done to help developers analyze and treat them.......  In this paper we propose to combine bug-finding rules with transformations that automatically introduce bug-fixes or workarounds when a possible bug is detected.  This work builds on our previous work on the Coccinelle tool, which targets device driver evolution....

Variability Bugs in Highly Configurable Systems: A Qualitative Analysis

DEFF Research Database (Denmark)

Abal, Iago; Melo, Jean; Stanciulescu, Stefan

2018-01-01

Variability-sensitive verification pursues effective analysis of the exponentially many variants in number of features of a program family. Several variability-aware techniques have been proposed, but researchers still lack examples of concrete bugs induced by variability, occurring in real large......-scale systems. A collection of real world bugs is needed to evaluate tool implementations of variability-sensitive analyses by testing them on real bugs. We present a qualitative study of 98 diverse variability bugs collected from bug-fixing commits in the Apache, BusyBox, Linux kernel and Marlin repositories....... We analyze each of the bugs, and record the results in a database. For each bug, we create a self-contained simplified C99 version and a simplified patch, in order to help researchers who are not experts on these subject studies to understand them, so that they can use it for evaluation...

Protecting Your Home from Bed Bugs

Science.gov (United States)

... your home: Inspect the luggage rack in your hotel room for bed bugs. Check secondhand furniture, beds, ... with Bed Bug Problems Discover. Accessibility EPA Administrator Budget & Performance Contracting Grants January 19, 2017 Web Snapshot ...

A theory for viral capsid assembly around electrostatic cores

Science.gov (United States)

Hagan, Michael F.

2009-03-01

We develop equilibrium and kinetic theories that describe the assembly of viral capsid proteins on a charged central core, as seen in recent experiments in which brome mosaic virus capsids assemble around nanoparticles functionalized with polyelectrolyte. We model interactions between capsid proteins and nanoparticle surfaces as the interaction of polyelectrolyte brushes with opposite charge using the nonlinear Poisson Boltzmann equation. The models predict that there is a threshold density of functionalized charge, above which capsids efficiently assemble around nanoparticles, and that light scatter intensity increases rapidly at early times without the lag phase characteristic of empty capsid assembly. These predictions are consistent with and enable interpretation of preliminary experimental data. However, the models predict a stronger dependence of nanoparticle incorporation efficiency on functionalized charge density than measured in experiments and do not completely capture a logarithmic growth phase seen in experimental light scatter. These discrepancies may suggest the presence of metastable disordered states in the experimental system. In addition to discussing future experiments for nanoparticle-capsid systems, we discuss broader implications for understanding assembly around charged cores such as nucleic acids.

The Cellular Chaperone Heat Shock Protein 90 Is Required for Foot-and-Mouth Disease Virus Capsid Precursor Processing and Assembly of Capsid Pentamers.

Science.gov (United States)

Newman, Joseph; Asfor, Amin S; Berryman, Stephen; Jackson, Terry; Curry, Stephen; Tuthill, Tobias J

2018-03-01

Productive picornavirus infection requires the hijacking of host cell pathways to aid with the different stages of virus entry, synthesis of the viral polyprotein, and viral genome replication. Many picornaviruses, including foot-and-mouth disease virus (FMDV), assemble capsids via the multimerization of several copies of a single capsid precursor protein into a pentameric subunit which further encapsidates the RNA. Pentamer formation is preceded by co- and posttranslational modification of the capsid precursor (P1-2A) by viral and cellular enzymes and the subsequent rearrangement of P1-2A into a structure amenable to pentamer formation. We have developed a cell-free system to study FMDV pentamer assembly using recombinantly expressed FMDV capsid precursor and 3C protease. Using this assay, we have shown that two structurally different inhibitors of the cellular chaperone heat shock protein 90 (hsp90) impeded FMDV capsid precursor processing and subsequent pentamer formation. Treatment of FMDV permissive cells with the hsp90 inhibitor prior to infection reduced the endpoint titer by more than 10-fold while not affecting the activity of a subgenomic replicon, indicating that translation and replication of viral RNA were unaffected by the drug. IMPORTANCE FMDV of the Picornaviridae family is a pathogen of huge economic importance to the livestock industry due to its effect on the restriction of livestock movement and necessary control measures required following an outbreak. The study of FMDV capsid assembly, and picornavirus capsid assembly more generally, has tended to be focused upon the formation of capsids from pentameric intermediates or the immediate cotranslational modification of the capsid precursor protein. Here, we describe a system to analyze the early stages of FMDV pentameric capsid intermediate assembly and demonstrate a novel requirement for the cellular chaperone hsp90 in the formation of these pentameric intermediates. We show the added complexity

Bed Bug Guidance for School Nurses

Science.gov (United States)

School nurses are often called upon to provide vital information to students, parents, teachers, and administrators. These tips on identifying, managing and preventing bed bugs will help you to effectively respond if bed bugs appear in your school.

Assassin bug uses aggressive mimicry to lure spider prey.

Science.gov (United States)

Wignall, Anne E; Taylor, Phillip W

2011-05-07

Assassin bugs (Stenolemus bituberus) hunt web-building spiders by invading the web and plucking the silk to generate vibrations that lure the resident spider into striking range. To test whether vibrations generated by bugs aggressively mimic the vibrations generated by insect prey, we compared the responses of spiders to bugs with how they responded to prey, courting male spiders and leaves falling into the web. We also analysed the associated vibrations. Similar spider orientation and approach behaviours were observed in response to vibrations from bugs and prey, whereas different behaviours were observed in response to vibrations from male spiders and leaves. Peak frequency and duration of vibrations generated by bugs were similar to those generated by prey and courting males. Further, vibrations from bugs had a temporal structure and amplitude that were similar to vibrations generated by leg and body movements of prey and distinctly different to vibrations from courting males or leaves, or prey beating their wings. To be an effective predator, bugs do not need to mimic the full range of prey vibrations. Instead bugs are general mimics of a subset of prey vibrations that fall within the range of vibrations classified by spiders as 'prey'.

Scaffold expulsion and genome packaging trigger stabilization of herpes simplex virus capsids

Science.gov (United States)

Roos, Wouter H.; Radtke, Kerstin; Kniesmeijer, Edward; Geertsema, Hylkje; Sodeik, Beate; Wuite, Gijs J. L.

2009-01-01

Herpes simplex virus type 1 (HSV1) capsids undergo extensive structural changes during maturation and DNA packaging. As a result, they become more stable and competent for nuclear egress. To further elucidate this stabilization process, we used biochemical and nanoindentation approaches to analyze the structural and mechanical properties of scaffold-containing (B), empty (A), and DNA-containing (C) nuclear capsids. Atomic force microscopy experiments revealed that A and C capsids were mechanically indistinguishable, indicating that the presence of DNA does not account for changes in mechanical properties during capsid maturation. Despite having the same rigidity, the scaffold-containing B capsids broke at significantly lower forces than A and C capsids. An extraction of pentons with guanidine hydrochloride (GuHCl) increased the flexibility of all capsids. Surprisingly, the breaking forces of the modified A and C capsids dropped to similar values as those of the GuHCl-treated B capsids, indicating that mechanical reinforcement occurs at the vertices. Nonetheless, it also showed that HSV1 capsids possess a remarkable structural integrity that was preserved after removal of pentons. We suggest that HSV1 capsids are stabilized after removal of the scaffold proteins, and that this stabilization is triggered by the packaging of DNA, but independent of the actual presence of DNA. PMID:19487681

Bug22p, a Conserved Centrosomal/Ciliary Protein Also Present in Higher Plants, Is Required for an Effective Ciliary Stroke in Paramecium ▿ †

Science.gov (United States)

Laligné, C.; Klotz, C.; Garreau de Loubresse, N.; Lemullois, M.; Hori, M.; Laurent, F. X.; Papon, J. F.; Louis, B.; Cohen, J.; Koll, F.

2010-01-01

Centrioles, cilia, and flagella are ancestral conserved organelles of eukaryotic cells. Among the proteins identified in the proteomics of ciliary proteins in Paramecium, we focus here on a protein, Bug22p, previously detected by cilia and basal-body high-throughput studies but never analyzed per se. Remarkably, this protein is also present in plants, which lack centrioles and cilia. Bug22p sequence alignments revealed consensus positions that distinguish species with centrioles/cilia from plants. In Paramecium, antibody and green fluorescent protein (GFP) fusion labeling localized Bug22p in basal bodies and cilia, and electron microscopy immunolabeling refined the localization to the terminal plate of the basal bodies, the transition zone, and spots along the axoneme, preferentially between the membrane and the microtubules. RNA interference (RNAi) depletion of Bug22p provoked a strong decrease in swimming speed, followed by cell death after a few days. High-speed video microscopy and morphological analysis of Bug22p-depleted cells showed that the protein plays an important role in the efficiency of ciliary movement by participating in the stroke shape and rigidity of cilia. The defects in cell swimming and growth provoked by RNAi can be complemented by expression of human Bug22p. This is the first reported case of complementation by a human gene in a ciliate. PMID:20118210

Antimicrobial peptide capsids of de novo design.

Science.gov (United States)

De Santis, Emiliana; Alkassem, Hasan; Lamarre, Baptiste; Faruqui, Nilofar; Bella, Angelo; Noble, James E; Micale, Nicola; Ray, Santanu; Burns, Jonathan R; Yon, Alexander R; Hoogenboom, Bart W; Ryadnov, Maxim G

2017-12-22

The spread of bacterial resistance to antibiotics poses the need for antimicrobial discovery. With traditional search paradigms being exhausted, approaches that are altogether different from antibiotics may offer promising and creative solutions. Here, we introduce a de novo peptide topology that-by emulating the virus architecture-assembles into discrete antimicrobial capsids. Using the combination of high-resolution and real-time imaging, we demonstrate that these artificial capsids assemble as 20-nm hollow shells that attack bacterial membranes and upon landing on phospholipid bilayers instantaneously (seconds) convert into rapidly expanding pores causing membrane lysis (minutes). The designed capsids show broad antimicrobial activities, thus executing one primary function-they destroy bacteria on contact.

Kissing Bug ( spp. Intrusion into Homes: Troublesome Bites and Domiciliation

Directory of Open Access Journals (Sweden)

Stephen A. Klotz

2016-01-01

Full Text Available Kissing bugs ( Triatoma spp. frequently enter homes and bite human and pet occupants. Bites may lead to severe allergic reactions and, in some cases, death. Kissing bugs are also vectors of Trypanosoma cruzi , the cause of Chagas disease. In general, modern houses in the United States are not conducive to domiciliation of kissing bugs (bugs living out their entire life within the home with the presence of eggs, nymphs, adults, and exuviae. Construction features such as concrete foundations, solid walls and ceilings, window screens, tight thresholds for doors and windows, and other measures impede bug entry into homes, and air conditioning reduces the need for open doors and windows. Where Chagas disease is endemic in Mexico and Central and South America, homes often have thatch roofs, adobe walls, and open doors and windows. We investigated numerous instances of kissing bug intrusions into homes in Southern Arizona, California, and Louisiana and documented the reactions to kissing bug bites. Our work confirms the importance of modern home construction in limiting kissing bug intrusions. Older homes, especially those lacking modern screening, caulking, and weather stripping to reduce air leakage, may be subject to kissing bug intrusions and domiciliation. We describe a community in Southern Arizona where domiciliation of homes by Triatoma recurva is common. We also provide recent data regarding kissing bug bites and allergic reactions to the bites.

Structural rigidity in the capsid assembly of cowpea chlorotic mottle virus

International Nuclear Information System (INIS)

Hespenheide, B M; Jacobs, D J; Thorpe, M F

2004-01-01

The cowpea chlorotic mottle virus (CCMV) has a protein cage, or capsid, which encloses its genetic material. The structure of the capsid consists of 180 copies of a single protein that self-assemble inside a cell to form a complete capsid with icosahedral symmetry. The icosahedral surface can be naturally divided into pentagonal and hexagonal faces, and the formation of either of these faces has been proposed to be the first step in the capsid assembly process. We have used the software FIRST to analyse the rigidity of pentameric and hexameric substructures of the complete capsid to explore the viability of certain capsid assembly pathways. FIRST uses the 3D pebble game to determine structural rigidity, and a brief description of this algorithm, as applied to body-bar networks, is given here. We find that the pentameric substructure, which corresponds to a pentagonal face on the icosahedral surface, provides the best structural properties for nucleating the capsid assembly process, consistent with experimental observations

Structural rigidity in the capsid assembly of cowpea chlorotic mottle virus

Science.gov (United States)

Hespenheide, B. M.; Jacobs, D. J.; Thorpe, M. F.

2004-11-01

The cowpea chlorotic mottle virus (CCMV) has a protein cage, or capsid, which encloses its genetic material. The structure of the capsid consists of 180 copies of a single protein that self-assemble inside a cell to form a complete capsid with icosahedral symmetry. The icosahedral surface can be naturally divided into pentagonal and hexagonal faces, and the formation of either of these faces has been proposed to be the first step in the capsid assembly process. We have used the software FIRST to analyse the rigidity of pentameric and hexameric substructures of the complete capsid to explore the viability of certain capsid assembly pathways. FIRST uses the 3D pebble game to determine structural rigidity, and a brief description of this algorithm, as applied to body-bar networks, is given here. We find that the pentameric substructure, which corresponds to a pentagonal face on the icosahedral surface, provides the best structural properties for nucleating the capsid assembly process, consistent with experimental observations.

Structural rigidity in the capsid assembly of cowpea chlorotic mottle virus

Energy Technology Data Exchange (ETDEWEB)

Hespenheide, B M [Department of Physics and Astronomy, Arizona State University, PO Box 871504, Tempe, AZ 85287-1504 (United States); Jacobs, D J [Department of Physics and Astronomy, California State University, 18111 Nordhoff Street, Northridge, CA 91330-8268 (United States); Thorpe, M F [Department of Physics and Astronomy, Arizona State University, PO Box 871504, Tempe, AZ 85287-1504 (United States)

2004-11-10

The cowpea chlorotic mottle virus (CCMV) has a protein cage, or capsid, which encloses its genetic material. The structure of the capsid consists of 180 copies of a single protein that self-assemble inside a cell to form a complete capsid with icosahedral symmetry. The icosahedral surface can be naturally divided into pentagonal and hexagonal faces, and the formation of either of these faces has been proposed to be the first step in the capsid assembly process. We have used the software FIRST to analyse the rigidity of pentameric and hexameric substructures of the complete capsid to explore the viability of certain capsid assembly pathways. FIRST uses the 3D pebble game to determine structural rigidity, and a brief description of this algorithm, as applied to body-bar networks, is given here. We find that the pentameric substructure, which corresponds to a pentagonal face on the icosahedral surface, provides the best structural properties for nucleating the capsid assembly process, consistent with experimental observations.

Ebselen, a Small-Molecule Capsid Inhibitor of HIV-1 Replication.

Science.gov (United States)

Thenin-Houssier, Suzie; de Vera, Ian Mitchelle S; Pedro-Rosa, Laura; Brady, Angela; Richard, Audrey; Konnick, Briana; Opp, Silvana; Buffone, Cindy; Fuhrmann, Jakob; Kota, Smitha; Billack, Blase; Pietka-Ottlik, Magdalena; Tellinghuisen, Timothy; Choe, Hyeryun; Spicer, Timothy; Scampavia, Louis; Diaz-Griffero, Felipe; Kojetin, Douglas J; Valente, Susana T

2016-04-01

The human immunodeficiency virus type 1 (HIV-1) capsid plays crucial roles in HIV-1 replication and thus represents an excellent drug target. We developed a high-throughput screening method based on a time-resolved fluorescence resonance energy transfer (HTS-TR-FRET) assay, using the C-terminal domain (CTD) of HIV-1 capsid to identify inhibitors of capsid dimerization. This assay was used to screen a library of pharmacologically active compounds, composed of 1,280in vivo-active drugs, and identified ebselen [2-phenyl-1,2-benzisoselenazol-3(2H)-one], an organoselenium compound, as an inhibitor of HIV-1 capsid CTD dimerization. Nuclear magnetic resonance (NMR) spectroscopic analysis confirmed the direct interaction of ebselen with the HIV-1 capsid CTD and dimer dissociation when ebselen is in 2-fold molar excess. Electrospray ionization mass spectrometry revealed that ebselen covalently binds the HIV-1 capsid CTD, likely via a selenylsulfide linkage with Cys198 and Cys218. This compound presents anti-HIV activity in single and multiple rounds of infection in permissive cell lines as well as in primary peripheral blood mononuclear cells. Ebselen inhibits early viral postentry events of the HIV-1 life cycle by impairing the incoming capsid uncoating process. This compound also blocks infection of other retroviruses, such as Moloney murine leukemia virus and simian immunodeficiency virus, but displays no inhibitory activity against hepatitis C and influenza viruses. This study reports the use of TR-FRET screening to successfully identify a novel capsid inhibitor, ebselen, validating HIV-1 capsid as a promising target for drug development. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

Mining Bug Databases for Unidentified Software Vulnerabilities

Energy Technology Data Exchange (ETDEWEB)

Dumidu Wijayasekara; Milos Manic; Jason Wright; Miles McQueen

2012-06-01

Identifying software vulnerabilities is becoming more important as critical and sensitive systems increasingly rely on complex software systems. It has been suggested in previous work that some bugs are only identified as vulnerabilities long after the bug has been made public. These vulnerabilities are known as hidden impact vulnerabilities. This paper discusses the feasibility and necessity to mine common publicly available bug databases for vulnerabilities that are yet to be identified. We present bug database analysis of two well known and frequently used software packages, namely Linux kernel and MySQL. It is shown that for both Linux and MySQL, a significant portion of vulnerabilities that were discovered for the time period from January 2006 to April 2011 were hidden impact vulnerabilities. It is also shown that the percentage of hidden impact vulnerabilities has increased in the last two years, for both software packages. We then propose an improved hidden impact vulnerability identification methodology based on text mining bug databases, and conclude by discussing a few potential problems faced by such a classifier.

Efficient Capsid Antigen Presentation From Adeno-Associated Virus Empty Virions In Vivo.

Science.gov (United States)

Pei, Xiaolei; Earley, Lauriel Freya; He, Yi; Chen, Xiaojing; Hall, Nikita Elexa; Samulski, Richard Jude; Li, Chengwen

2018-01-01

Adeno-associated virus (AAV) vectors have been successfully applied in clinical trials for hemophilic patients. Although promising, the clinical results suggest that the capsid-specific CD8+T cell response has a negative effect on therapeutic success. In an in vitro analysis using an engineered AAV virus carrying immune-dominant SIINFEKL peptide in the capsid backbone, we have previously demonstrated that capsid antigen presentation from full (genome containing) AAV capsids requires endosome escape and is proteasome dependent and that no capsid antigen presentation is induced from empty virions. In the present study, we examined capsid antigen presentation from administration of empty virions in animal models. In wild-type mice, similar to AAV full particles, capsid antigen presentation from AAV empty virion infection was dose dependent, and the kinetics studies showed that antigen presentation was detected from 2 to 40 days after AAV empty virion administration. In the transporter associated with antigen processing 1 deficient (TAP-/-) mice, capsid antigen presentation was inhibited from both AAV full and empty virions, but higher inhibition was achieved from AAV full particle administration than that from empty virions. This indicates that the pathway of capsid antigen presentation from AAV transduction is dependent on proteasome-mediated degradation of AAV capsids (mainly for full particles) and that the endosomal pathway may also play a role in antigen presentation from empty particles but not full virions. The capsid antigen presentation efficiency from AAV preparations was positively correlated with the amount of empty virions contaminated with full particles. Collectively, the results indicate that contamination of AAV empty virions induces efficient antigen presentation in vivo and the mechanism of capsid antigen presentation from empty virions involves both endosomal and proteasomal pathways. The elucidation of capsid antigen presentation from AAV empty

Bug City: Crickets, Grasshoppers & Friends [Videotape].

Science.gov (United States)

1998

"Bug City" is a video series created to help children (grades 1-6) learn about insects and other small critters. All aspects of bug life are touched upon, including body structure, food, habitat, life cycle, mating habits, camouflage, mutualism (symbiosis), adaptations, social behavior, and more. Each program features dramatic…

Bug City: Spiders and Scorpions [Videotape].

Science.gov (United States)

1998

"Bug City" is a video series created to help children learn about insects and other small critters. All aspects of bug life are touched upon including body structure, food, habitat, life cycle, mating habits, camouflage, mutualism (symbiosis), adaptations, social behavior, and more. Each program features dramatic microscopic photography,…

Bug City: Butterflies and Moths [Videotape].

Science.gov (United States)

1998

"Bug City" is a video series created to help children learn about insects and other small critters. All aspects of bug life are touched upon including body structure, food, habitat, life cycle, mating habits, camouflage, mutualism (symbiosis), adaptations, social behavior, and more. Each program features dramatic microscopic photography,…

Mutation of the N-Terminal Region of Chikungunya Virus Capsid Protein: Implications for Vaccine Design.

Science.gov (United States)

Taylor, Adam; Liu, Xiang; Zaid, Ali; Goh, Lucas Y H; Hobson-Peters, Jody; Hall, Roy A; Merits, Andres; Mahalingam, Suresh

2017-02-21

Mosquito-transmitted chikungunya virus (CHIKV) is an arthritogenic alphavirus of the Togaviridae family responsible for frequent outbreaks of arthritic disease in humans. Capsid protein, a structural protein encoded by the CHIKV RNA genome, is able to translocate to the host cell nucleolus. In encephalitic alphaviruses, nuclear translocation induces host cell transcriptional shutoff; however, the role of capsid protein nucleolar localization in arthritogenic alphaviruses remains unclear. Using recombinant enhanced green fluorescent protein (EGFP)-tagged expression constructs and CHIKV infectious clones, we describe a nucleolar localization sequence (NoLS) in the N-terminal region of capsid protein, previously uncharacterized in CHIKV. Mutation of the NoLS by site-directed mutagenesis reduced efficiency of nuclear import of CHIKV capsid protein. In the virus, mutation of the capsid protein NoLS (CHIKV-NoLS) attenuated replication in mammalian and mosquito cells, producing a small-plaque phenotype. Attenuation of CHIKV-NoLS is likely due to disruption of the viral replication cycle downstream of viral RNA synthesis. In mice, CHIKV-NoLS infection caused no disease signs compared to wild-type CHIKV (CHIKV-WT)-infected mice; lack of disease signs correlated with significantly reduced viremia and decreased expression of proinflammatory factors. Mice immunized with CHIKV-NoLS, challenged with CHIKV-WT at 30 days postimmunization, develop no disease signs and no detectable viremia. Serum from CHIKV-NoLS-immunized mice is able to efficiently neutralize CHIKV infection in vitro Additionally, CHIKV-NoLS-immunized mice challenged with the related alphavirus Ross River virus showed reduced early and peak viremia postchallenge, indicating a cross-protective effect. The high degree of CHIKV-NoLS attenuation may improve CHIKV antiviral and rational vaccine design. IMPORTANCE CHIKV is a mosquito-borne pathogen capable of causing explosive epidemics of incapacitating joint pain

Machine Learning or Information Retrieval Techniques for Bug Triaging: Which is better?

Directory of Open Access Journals (Sweden)

Anjali Goyal

2017-07-01

Full Text Available Bugs are the inevitable part of a software system. Nowadays, large software development projects even release beta versions of their products to gather bug reports from users. The collected bug reports are then worked upon by various developers in order to resolve the defects and make the final software product more reliable. The high frequency of incoming bugs makes the bug handling a difficult and time consuming task. Bug assignment is an integral part of bug triaging that aims at the process of assigning a suitable developer for the reported bug who corrects the source code in order to resolve the bug. There are various semi and fully automated techniques to ease the task of bug assignment. This paper presents the current state of the art of various techniques used for bug report assignment. Through exhaustive research, the authors have observed that machine learning and information retrieval based bug assignment approaches are most popular in literature. A deeper investigation has shown that the trend of techniques is taking a shift from machine learning based approaches towards information retrieval based approaches. Therefore, the focus of this work is to find the reason behind the observed drift and thus a comparative analysis is conducted on the bug reports of the Mozilla, Eclipse, Gnome and Open Office projects in the Bugzilla repository. The results of the study show that the information retrieval based technique yields better efficiency in recommending the developers for bug reports.

Effects of Point Mutations in the Major Capsid Protein of Beet Western Yellows Virus on Capsid Formation, Virus Accumulation, and Aphid Transmission

Science.gov (United States)

Brault, V.; Bergdoll, M.; Mutterer, J.; Prasad, V.; Pfeffer, S.; Erdinger, M.; Richards, K. E.; Ziegler-Graff, V.

2003-01-01

Point mutations were introduced into the major capsid protein (P3) of cloned infectious cDNA of the polerovirus beet western yellows virus (BWYV) by manipulation of cloned infectious cDNA. Seven mutations targeted sites on the S domain predicted to lie on the capsid surface. An eighth mutation eliminated two arginine residues in the R domain, which is thought to extend into the capsid interior. The effects of the mutations on virus capsid formation, virus accumulation in protoplasts and plants, and aphid transmission were tested. All of the mutants replicated in protoplasts. The S-domain mutant W166R failed to protect viral RNA from RNase attack, suggesting that this particular mutation interfered with stable capsid formation. The R-domain mutant R7A/R8A protected ∼90% of the viral RNA strand from RNase, suggesting that lower positive-charge density in the mutant capsid interior interfered with stable packaging of the complete strand into virions. Neither of these mutants systemically infected plants. The six remaining mutants properly packaged viral RNA and could invade Nicotiana clevelandii systemically following agroinfection. Mutant Q121E/N122D was poorly transmitted by aphids, implicating one or both targeted residues in virus-vector interactions. Successful transmission of mutant D172N was accompanied either by reversion to the wild type or by appearance of a second-site mutation, N137D. This finding indicates that D172 is also important for transmission but that the D172N transmission defect can be compensated for by a “reverse” substitution at another site. The results have been used to evaluate possible structural models for the BWYV capsid. PMID:12584348

NRFixer: Sentiment Based Model for Predicting the Fixability of Non-Reproducible Bugs

Directory of Open Access Journals (Sweden)

Anjali Goyal

2017-08-01

Full Text Available Software maintenance is an essential step in software development life cycle. Nowadays, software companies spend approximately 45\\% of total cost in maintenance activities. Large software projects maintain bug repositories to collect, organize and resolve bug reports. Sometimes it is difficult to reproduce the reported bug with the information present in a bug report and thus this bug is marked with resolution non-reproducible (NR. When NR bugs are reconsidered, a few of them might get fixed (NR-to-fix leaving the others with the same resolution (NR. To analyse the behaviour of developers towards NR-to-fix and NR bugs, the sentiment analysis of NR bug report textual contents has been conducted. The sentiment analysis of bug reports shows that NR bugs' sentiments incline towards more negativity than reproducible bugs. Also, there is a noticeable opinion drift found in the sentiments of NR-to-fix bug reports. Observations driven from this analysis were an inspiration to develop a model that can judge the fixability of NR bugs. Thus a framework, {NRFixer,} which predicts the probability of NR bug fixation, is proposed. {NRFixer} was evaluated with two dimensions. The first dimension considers meta-fields of bug reports (model-1 and the other dimension additionally incorporates the sentiments (model-2 of developers for prediction. Both models were compared using various machine learning classifiers (Zero-R, naive Bayes, J48, random tree and random forest. The bug reports of Firefox and Eclipse projects were used to test {NRFixer}. In Firefox and Eclipse projects, J48 and Naive Bayes classifiers achieve the best prediction accuracy, respectively. It was observed that the inclusion of sentiments in the prediction model shows a rise in the prediction accuracy ranging from 2 to 5\\% for various classifiers.

Parvovirus Capsid Structures Required for Infection: Mutations Controlling Receptor Recognition and Protease Cleavages.

Science.gov (United States)

Callaway, Heather M; Feng, Kurtis H; Lee, Donald W; Allison, Andrew B; Pinard, Melissa; McKenna, Robert; Agbandje-McKenna, Mavis; Hafenstein, Susan; Parrish, Colin R

2017-01-15

Parvovirus capsids are small but complex molecular machines responsible for undertaking many of the steps of cell infection, genome packing, and cell-to-cell as well as host-to-host transfer. The details of parvovirus infection of cells are still not fully understood, but the processes must involve small changes in the capsid structure that allow the endocytosed virus to escape from the endosome, pass through the cell cytoplasm, and deliver the single-stranded DNA (ssDNA) genome to the nucleus, where viral replication occurs. Here, we examine capsid substitutions that eliminate canine parvovirus (CPV) infectivity and identify how those mutations changed the capsid structure or altered interactions with the infectious pathway. Amino acid substitutions on the exterior surface of the capsid (Gly299Lys/Ala300Lys) altered the binding of the capsid to transferrin receptor type 1 (TfR), particularly during virus dissociation from the receptor, but still allowed efficient entry into both feline and canine cells without successful infection. These substitutions likely control specific capsid structural changes resulting from TfR binding required for infection. A second set of changes on the interior surface of the capsid reduced viral infectivity by >100-fold and included two cysteine residues and neighboring residues. One of these substitutions, Cys270Ser, modulates a VP2 cleavage event found in ∼10% of the capsid proteins that also was shown to alter capsid stability. A neighboring substitution, Pro272Lys, significantly reduced capsid assembly, while a Cys273Ser change appeared to alter capsid transport from the nucleus. These mutants reveal additional structural details that explain cell infection processes of parvovirus capsids. Parvoviruses are commonly found in both vertebrate and invertebrate animals and cause widespread disease. They are also being developed as oncolytic therapeutics and as gene therapy vectors. Most functions involved in infection or transduction

Parvovirus Capsid Structures Required for Infection: Mutations Controlling Receptor Recognition and Protease Cleavages

Science.gov (United States)

Callaway, Heather M.; Feng, Kurtis H.; Lee, Donald W.; Pinard, Melissa; McKenna, Robert; Agbandje-McKenna, Mavis; Hafenstein, Susan

2016-01-01

ABSTRACT Parvovirus capsids are small but complex molecular machines responsible for undertaking many of the steps of cell infection, genome packing, and cell-to-cell as well as host-to-host transfer. The details of parvovirus infection of cells are still not fully understood, but the processes must involve small changes in the capsid structure that allow the endocytosed virus to escape from the endosome, pass through the cell cytoplasm, and deliver the single-stranded DNA (ssDNA) genome to the nucleus, where viral replication occurs. Here, we examine capsid substitutions that eliminate canine parvovirus (CPV) infectivity and identify how those mutations changed the capsid structure or altered interactions with the infectious pathway. Amino acid substitutions on the exterior surface of the capsid (Gly299Lys/Ala300Lys) altered the binding of the capsid to transferrin receptor type 1 (TfR), particularly during virus dissociation from the receptor, but still allowed efficient entry into both feline and canine cells without successful infection. These substitutions likely control specific capsid structural changes resulting from TfR binding required for infection. A second set of changes on the interior surface of the capsid reduced viral infectivity by >100-fold and included two cysteine residues and neighboring residues. One of these substitutions, Cys270Ser, modulates a VP2 cleavage event found in ∼10% of the capsid proteins that also was shown to alter capsid stability. A neighboring substitution, Pro272Lys, significantly reduced capsid assembly, while a Cys273Ser change appeared to alter capsid transport from the nucleus. These mutants reveal additional structural details that explain cell infection processes of parvovirus capsids. IMPORTANCE Parvoviruses are commonly found in both vertebrate and invertebrate animals and cause widespread disease. They are also being developed as oncolytic therapeutics and as gene therapy vectors. Most functions involved in

All-atom molecular dynamics calculation study of entire poliovirus empty capsids in solution

Energy Technology Data Exchange (ETDEWEB)

Andoh, Y.; Yoshii, N.; Yamada, A.; Kojima, H.; Mizutani, K.; Okazaki, S., E-mail: okazaki@apchem.nagoya-u.ac.jp [Department of Applied Chemistry, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603 (Japan); Fujimoto, K. [Department of Pharmacy, College of Pharmaceutical Sciences, Ritsumeikan University, Nojihigashi, Kusatsu, Shiga 525-8577 (Japan); Nakagawa, A. [Institute for Protein Research, Osaka University, Yamadaoka, Suita, Osaka 565-0871 (Japan); Nomoto, A. [Institute of Microbial Chemistry, Kamiosaki, Shinagawa-ku, Tokyo 141-0021 (Japan)

2014-10-28

Small viruses that belong, for example, to the Picornaviridae, such as poliovirus and foot-and-mouth disease virus, consist simply of capsid proteins and a single-stranded RNA (ssRNA) genome. The capsids are quite stable in solution to protect the genome from the environment. Here, based on long-time and large-scale 6.5 × 10{sup 6} all-atom molecular dynamics calculations for the Mahoney strain of poliovirus, we show microscopic properties of the viral capsids at a molecular level. First, we found equilibrium rapid exchange of water molecules across the capsid. The exchange rate is so high that all water molecules inside the capsid (about 200 000) can leave the capsid and be replaced by water molecules from the outside in about 25 μs. This explains the capsid's tolerance to high pressures and deactivation by exsiccation. In contrast, the capsid did not exchange ions, at least within the present simulation time of 200 ns. This implies that the capsid can function, in principle, as a semipermeable membrane. We also found that, similar to the xylem of trees, the pressure of the solution inside the capsid without the genome was negative. This is caused by coulombic interaction of the solution inside the capsid with the capsid excess charges. The negative pressure may be compensated by positive osmotic pressure by the solution-soluble ssRNA and the counter ions introduced into it.

All-atom molecular dynamics calculation study of entire poliovirus empty capsids in solution

International Nuclear Information System (INIS)

Andoh, Y.; Yoshii, N.; Yamada, A.; Kojima, H.; Mizutani, K.; Okazaki, S.; Fujimoto, K.; Nakagawa, A.; Nomoto, A.

2014-01-01

Small viruses that belong, for example, to the Picornaviridae, such as poliovirus and foot-and-mouth disease virus, consist simply of capsid proteins and a single-stranded RNA (ssRNA) genome. The capsids are quite stable in solution to protect the genome from the environment. Here, based on long-time and large-scale 6.5 × 10 6 all-atom molecular dynamics calculations for the Mahoney strain of poliovirus, we show microscopic properties of the viral capsids at a molecular level. First, we found equilibrium rapid exchange of water molecules across the capsid. The exchange rate is so high that all water molecules inside the capsid (about 200 000) can leave the capsid and be replaced by water molecules from the outside in about 25 μs. This explains the capsid's tolerance to high pressures and deactivation by exsiccation. In contrast, the capsid did not exchange ions, at least within the present simulation time of 200 ns. This implies that the capsid can function, in principle, as a semipermeable membrane. We also found that, similar to the xylem of trees, the pressure of the solution inside the capsid without the genome was negative. This is caused by coulombic interaction of the solution inside the capsid with the capsid excess charges. The negative pressure may be compensated by positive osmotic pressure by the solution-soluble ssRNA and the counter ions introduced into it

All-atom molecular dynamics calculation study of entire poliovirus empty capsids in solution

Science.gov (United States)

Andoh, Y.; Yoshii, N.; Yamada, A.; Fujimoto, K.; Kojima, H.; Mizutani, K.; Nakagawa, A.; Nomoto, A.; Okazaki, S.

2014-10-01

Small viruses that belong, for example, to the Picornaviridae, such as poliovirus and foot-and-mouth disease virus, consist simply of capsid proteins and a single-stranded RNA (ssRNA) genome. The capsids are quite stable in solution to protect the genome from the environment. Here, based on long-time and large-scale 6.5 × 106 all-atom molecular dynamics calculations for the Mahoney strain of poliovirus, we show microscopic properties of the viral capsids at a molecular level. First, we found equilibrium rapid exchange of water molecules across the capsid. The exchange rate is so high that all water molecules inside the capsid (about 200 000) can leave the capsid and be replaced by water molecules from the outside in about 25 μs. This explains the capsid's tolerance to high pressures and deactivation by exsiccation. In contrast, the capsid did not exchange ions, at least within the present simulation time of 200 ns. This implies that the capsid can function, in principle, as a semipermeable membrane. We also found that, similar to the xylem of trees, the pressure of the solution inside the capsid without the genome was negative. This is caused by coulombic interaction of the solution inside the capsid with the capsid excess charges. The negative pressure may be compensated by positive osmotic pressure by the solution-soluble ssRNA and the counter ions introduced into it.

Kissing Bug (Triatoma spp.) Intrusion into Homes: Troublesome Bites and Domiciliation.

Science.gov (United States)

Klotz, Stephen A; Shirazi, F Mazda; Boesen, Keith; Beatty, Norman L; Dorn, Patricia L; Smith, Shannon; Schmidt, Justin O

2016-01-01

Kissing bugs (Triatoma spp.) frequently enter homes and bite human and pet occupants. Bites may lead to severe allergic reactions and, in some cases, death. Kissing bugs are also vectors of Trypanosoma cruzi, the cause of Chagas disease. In general, modern houses in the United States are not conducive to domiciliation of kissing bugs (bugs living out their entire life within the home with the presence of eggs, nymphs, adults, and exuviae). Construction features such as concrete foundations, solid walls and ceilings, window screens, tight thresholds for doors and windows, and other measures impede bug entry into homes, and air conditioning reduces the need for open doors and windows. Where Chagas disease is endemic in Mexico and Central and South America, homes often have thatch roofs, adobe walls, and open doors and windows. We investigated numerous instances of kissing bug intrusions into homes in Southern Arizona, California, and Louisiana and documented the reactions to kissing bug bites. Our work confirms the importance of modern home construction in limiting kissing bug intrusions. Older homes, especially those lacking modern screening, caulking, and weather stripping to reduce air leakage, may be subject to kissing bug intrusions and domiciliation. We describe a community in Southern Arizona where domiciliation of homes by Triatoma recurva is common. We also provide recent data regarding kissing bug bites and allergic reactions to the bites.

Print a Bed Bug Card - (Single Cards)

Science.gov (United States)

Two sets of business-card-sized lists of tips for recognizing bed bugs and the signs of an infestation, including a photo of bed bugs to assist identification. One card is for general use around home or office, the other for travelers.

Varicella-zoster virus induces the formation of dynamic nuclear capsid aggregates

Energy Technology Data Exchange (ETDEWEB)

Lebrun, Marielle [University of Liege (ULg), GIGA-Infection Immunity and Inflammation, Laboratory of Virology and Immunology, Liege (Belgium); Thelen, Nicolas; Thiry, Marc [University of Liege (ULg), GIGA-Neurosciences, Laboratory of Cellular and Tissular Biology, Liege (Belgium); Riva, Laura; Ote, Isabelle; Condé, Claude; Vandevenne, Patricia [University of Liege (ULg), GIGA-Infection Immunity and Inflammation, Laboratory of Virology and Immunology, Liege (Belgium); Di Valentin, Emmanuel [University of Liege (ULg), GIGA-Viral Vectors Platform, Liege (Belgium); Bontems, Sébastien [University of Liege (ULg), GIGA-Infection Immunity and Inflammation, Laboratory of Virology and Immunology, Liege (Belgium); Sadzot-Delvaux, Catherine, E-mail: csadzot@ulg.ac.be [University of Liege (ULg), GIGA-Infection Immunity and Inflammation, Laboratory of Virology and Immunology, Liege (Belgium)

2014-04-15

The first step of herpesviruses virion assembly occurs in the nucleus. However, the exact site where nucleocapsids are assembled, where the genome and the inner tegument are acquired, remains controversial. We created a recombinant VZV expressing ORF23 (homologous to HSV-1 VP26) fused to the eGFP and dually fluorescent viruses with a tegument protein additionally fused to a red tag (ORF9, ORF21 and ORF22 corresponding to HSV-1 UL49, UL37 and UL36). We identified nuclear dense structures containing the major capsid protein, the scaffold protein and maturing protease, as well as ORF21 and ORF22. Correlative microscopy demonstrated that the structures correspond to capsid aggregates and time-lapse video imaging showed that they appear prior to the accumulation of cytoplasmic capsids, presumably undergoing the secondary egress, and are highly dynamic. Our observations suggest that these structures might represent a nuclear area important for capsid assembly and/or maturation before the budding at the inner nuclear membrane. - Highlights: • We created a recombinant VZV expressing the small capsid protein fused to the eGFP. • We identified nuclear dense structures containing capsid and procapsid proteins. • Correlative microscopy showed that the structures correspond to capsid aggregates. • Procapsids and partial capsids are found within the aggregates of WT and eGFP-23 VZV. • FRAP and FLIP experiments demonstrated that they are dynamic structures.

Finding error handling bugs in OpenSSL using Coccinelle

DEFF Research Database (Denmark)

Lawall, Julia; Laurie, Ben; Hansen, René Rydhof

2010-01-01

OpenSSL is a library providing various functionalities relating to secure network communication.  Detecting and fixing bugs in OpenSSL code is thus essential, particularly when such bugs can lead to malicious attacks.  In previous work, we have proposed a methodology for finding API usage protocols...... in Linux kernel code using the program matching and transformation engine Coccinelle.  In this work, we report on our experience in applying this methodology to OpenSSL, focusing on API usage protocols related to error handling.  We have detected over 30 bugs in a recent OpenSSL snapshot, and in many cases...... it was possible to correct the bugs automatically.  Our patches correcting these bugs have been accepted by the OpenSSL developers.  This work furthermore confirms the applicability of our methodology to user-level code....

Reactive oxygen species promote heat shock protein 90-mediated HBV capsid assembly

International Nuclear Information System (INIS)

Kim, Yoon Sik; Seo, Hyun Wook; Jung, Guhung

2015-01-01

Hepatitis B virus (HBV) infection induces reactive oxygen species (ROS) production and has been associated with the development of hepatocellular carcinoma (HCC). ROS are also an important factor in HCC because the accumulated ROS leads to abnormal cell proliferation and chromosome mutation. In oxidative stress, heat shock protein 90 (Hsp90) and glutathione (GSH) function as part of the defense mechanism. Hsp90 prevents cellular component from oxidative stress, and GSH acts as antioxidants scavenging ROS in the cell. However, it is not known whether molecules regulated by oxidative stress are involved in HBV capsid assembly. Based on the previous study that Hsp90 facilitates HBV capsid assembly, which is an important step for the packing of viral particles, here, we show that ROS enrich Hsp90-driven HBV capsid formation. In cell-free system, HBV capsid assembly was facilitated by ROS with Hsp90, whereas it was decreased without Hsp90. In addition, GSH inhibited the function of Hsp90 to decrease HBV capsid assembly. Consistent with the result of cell-free system, ROS and buthionine sulfoximine (BS), an inhibitor of GSH synthesis, increased HBV capsid formation in HepG2.2.15 cells. Thus, our study uncovers the interplay between ROS and Hsp90 during HBV capsid assembly. - Highlights: • We examined H 2 O 2 and GSH modulate HBV capsid assembly. • H 2 O 2 facilitates HBV capsid assembly in the presence of Hsp90. • GSH inhibits function of Hsp90 in facilitating HBV capsid assembly. • H 2 O 2 and GSH induce conformation change of Hsp90

Reactive oxygen species promote heat shock protein 90-mediated HBV capsid assembly

Energy Technology Data Exchange (ETDEWEB)

Kim, Yoon Sik, E-mail: yumshak@naver.com; Seo, Hyun Wook, E-mail: suruk@naver.com; Jung, Guhung, E-mail: drjung@snu.ac.kr

2015-02-13

Hepatitis B virus (HBV) infection induces reactive oxygen species (ROS) production and has been associated with the development of hepatocellular carcinoma (HCC). ROS are also an important factor in HCC because the accumulated ROS leads to abnormal cell proliferation and chromosome mutation. In oxidative stress, heat shock protein 90 (Hsp90) and glutathione (GSH) function as part of the defense mechanism. Hsp90 prevents cellular component from oxidative stress, and GSH acts as antioxidants scavenging ROS in the cell. However, it is not known whether molecules regulated by oxidative stress are involved in HBV capsid assembly. Based on the previous study that Hsp90 facilitates HBV capsid assembly, which is an important step for the packing of viral particles, here, we show that ROS enrich Hsp90-driven HBV capsid formation. In cell-free system, HBV capsid assembly was facilitated by ROS with Hsp90, whereas it was decreased without Hsp90. In addition, GSH inhibited the function of Hsp90 to decrease HBV capsid assembly. Consistent with the result of cell-free system, ROS and buthionine sulfoximine (BS), an inhibitor of GSH synthesis, increased HBV capsid formation in HepG2.2.15 cells. Thus, our study uncovers the interplay between ROS and Hsp90 during HBV capsid assembly. - Highlights: • We examined H{sub 2}O{sub 2} and GSH modulate HBV capsid assembly. • H{sub 2}O{sub 2} facilitates HBV capsid assembly in the presence of Hsp90. • GSH inhibits function of Hsp90 in facilitating HBV capsid assembly. • H{sub 2}O{sub 2} and GSH induce conformation change of Hsp90.

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.

Bug City: House and Backyard Insects [Videotape].

Science.gov (United States)

1998

"Bug City" is a video series created to help children learn about insects and other small critters. All aspects of bug life are touched upon including body structure, food, habitat, life cycle, mating habits, camouflage, mutualism (symbiosis), adaptations, social behavior, and more. Each program features dramatic microscopic photography,…

Insecticide susceptibility of the green plant bug, Apolygus lucorum Meyer-Dür (Homoptera: Miridae and two predatory arthropods

Directory of Open Access Journals (Sweden)

Zhang Zhengqun

2015-12-01

Full Text Available The green plant bug (Apolygus lucorum Meyer-Dür is a key pest of Bt cotton in China. Along with biological control, chemical control is one of the most important strategies in A. lucorum Integrated Pest Management (IPM. The goal of this study was to evaluate the toxicity of eight conventional insecticides to A. lucorum and to assess the susceptibility of two generalist predators Chrysopa sinica (Jieder and Propylaea japonica (Thunbery to insecticides that are commonly used in A. lucorum management. Via glass-vial and leaf-dip bioassay, toxicity tests with selected insecticides at two different life-stages of A. lucorum indicated significant differences between the LD50 or LC50 values for these compounds within different insecticidal classes. Phenylpyrazole fipronil had the highest toxicity to 4th-instar nymphs and adults of A. lucorum, whereas neonicotinoid imidacloprid had the lowest toxicity among the insecticides. Females were more tolerant to insecticides than were males, as shown by the higher LD50 values for females. Furthermore, laboratory tests showed that endosulfan had the highest selectivity to C. sinica and P. japonica: the selective toxicity ratios (STRs were superior to other tested insecticides, particularly imidacloprid, and were 5.396 and 4.749-fold higher than baseline STRs, respectively. From this study, we conclude that fipronil can potentially be used to efficiently control A. lucorum. An alternative control agent worth consideration is endosulfan, owing to its relative safety to non-targeted natural enemies.

Bed Bugs FAQs

Science.gov (United States)

... Europe. Bed bugs have been found in five-star hotels and resorts and their presence is not ... Health – Division of Parasitic Diseases Email Recommend Tweet YouTube Instagram Listen Watch RSS ABOUT About CDC Jobs ...

Identification of Factors Promoting HBV Capsid Self-Assembly by Assembly-Promoting Antivirals.

Science.gov (United States)

Rath, Soumya Lipsa; Liu, Huihui; Okazaki, Susumu; Shinoda, Wataru

2018-02-26

Around 270 million individuals currently live with hepatitis B virus (HBV) infection. Heteroaryldihydropyrimidines (HAPs) are a family of antivirals that target the HBV capsid protein and induce aberrant self-assembly. The capsids formed resemble the native capsid structure but are unable to propagate the virus progeny because of a lack of RNA/DNA. Under normal conditions, self-assembly is initiated by the viral genome. The mode of action of HAPs, however, remains largely unknown. In this work, using molecular dynamics simulations, we attempted to understand the action of HAP by comparing the dynamics of capsid proteins with and without HAPs. We found that the inhibitor is more stable in higher oligomers. It retains its stability in the hexamer throughout 1 μs of simulation. Our results also show that the inhibitor might help in stabilizing the C-terminus, the HBc 149-183 arginine-rich domain of the capsid protein. The C-termini of dimers interact with each other, assisted by the HAP inhibitor. During capsid assembly, the termini are supposed to directly interact with the viral genome, thereby suggesting that the viral genome might work in a similar way to stabilize the capsid protein. Our results may help in understanding the underlying molecular mechanism of HBV capsid self-assembly, which should be crucial for exploring new drug targets and structure-based drug design.

Comparison of management strategies for squash bugs (Hemiptera: Coreidae) in watermelon.

Science.gov (United States)

Dogramaci, Mahmut; Shrefler, James W; Roberts, B Warren; Pair, Sam; Edelson, J V

2004-12-01

Two watermelon pest management practices, a squash trap crop and a standard recommendation using soil-applied carbofuran, were compared using large-scale field plots to assess trap crop suitability as a replacement for the standard in 2000, 2001, and 2002. In both systems, foliar insecticide applications were used to control squash bugs when populations exceeded threshold levels. During 2001 and 2002, a treatment of untreated watermelon was used. Early season adult insects, from seedling to fruit set, are most critical for watermelon. Significantly fewer early adult bugs were found on watermelon in the trap crop than in the standard recommended practice in 1 of 3 yr. In both years, significantly fewer adult squash bugs were found in watermelon in the trap crop than in untreated fields. The standard recommended practice significantly reduced adult squash bugs in watermelon compared with the untreated in 1 of 2 yr. There was no significant correlation of watermelon yield and squash bug density, indicating that squash bug densities were too low to impact yield. Although squash bugs were reduced significantly by the trap crop, marketable watermelon yields were lower in the squash trap crop than in untreated watermelon, suggesting that pest management treatments may interfere with crop productivity factors other than squash bug colonization. Results suggest that mid-season production squash bug should be managed by monitoring populations and using insecticides as needed rather than using at-plant treatment. Further research is needed to compare treatments during early-season production.

Effective Bug Finding in C Programs with Shape and Effect Abstractions

DEFF Research Database (Denmark)

Abal, Iago; Brabrand, Claus; Wasowski, Andrzej

2017-01-01

Software projects tend to suffer from conceptually simple resource manipulation bugs, such as accessing a de-allocated memory region, or acquiring a non-reentrant lock twice. Static code scanners are used extensively to remove these bugs from projects like the Linux kernel. Yet, when the manipula......Software projects tend to suffer from conceptually simple resource manipulation bugs, such as accessing a de-allocated memory region, or acquiring a non-reentrant lock twice. Static code scanners are used extensively to remove these bugs from projects like the Linux kernel. Yet, when...... the Linux kernel. Our results show that our tool is more effective at finding bugs than similar code-scanning tools. EBA analyzes the drivers/ directory of Linux (nine thousand files) in less than thirty minutes, and uncovers a handful previously unknown double-lock bugs in various drivers....

The odorant receptor co-receptor from the bed bug, Cimex lectularius L.

Directory of Open Access Journals (Sweden)

Immo A Hansen

Full Text Available Recently, the bed bug, Cimex lectularius L. has re-emerged as a serious and growing problem in many parts of the world. Presence of resistant bed bugs and the difficulty to eliminate them has renewed interest in alternative control tactics. Similar to other haematophagous arthropods, bed bugs rely on their olfactory system to detect semiochemicals in the environment. Previous studies have morphologically characterized olfactory organs of bed bugs' antenna and have physiologically evaluated the responses of olfactory receptor neurons (ORNs to host-derived chemicals. To date, odorant binding proteins (OBPs and odorant receptors (ORs associated with these olfaction processes have not been studied in bed bugs. Chemoreception in insects requires formation of heteromeric complexes of ORs and a universal OR coreceptor (Orco. Orco is the constant chain of every odorant receptor in insects and is critical for insect olfaction but does not directly bind to odorants. Orco agonists and antagonists have been suggested as high-value targets for the development of novel insect repellents. In this study, we have performed RNAseq of bed bug sensory organs and identified several odorant receptors as well as Orco. We characterized Orco expression and investigated the effect of chemicals targeting Orco on bed bug behavior and reproduction. We have identified partial cDNAs of six C. lectularius OBPs and 16 ORs. Full length bed bug Orco was cloned and sequenced. Orco is widely expressed in different parts of the bed bug including OR neurons and spermatozoa. Treatment of bed bugs with the agonist VUAA1 changed bed bug pheromone-induced aggregation behavior and inactivated spermatozoa. We have described and characterized for the first time OBPs, ORs and Orco in bed bugs. Given the importance of these molecules in chemoreception of this insect they are interesting targets for the development of novel insect behavior modifiers.

Between-group transmission dynamics of the swallow bug, Oeciacus vicarius.

Science.gov (United States)

Brown, Charles R; Brown, Mary Bomberger

2005-06-01

The parasitic cimicid swallow bug, Oeciacus vicarius, is the principal invertebrate vector for Buggy Creek virus (BCRV) and has also been associated with Venezuelan equine encephalitis virus. To help understand the spread of this vector, we experimentally measured the transmission of O. vicarius between groups (colonies) of its main host, the cliff swallow (Petrochelidonpyrrhonota), in the field. Transmission of bugs between colonies varied significantly with year, size of the colony, and week within the season. Bug immigration into sites tended to peak in mid-summer. Swallow nests in larger colonies had more consistent rates of bug introduction than did nests in small colonies, but within a colony a given nest's weekly immigrant-bug count varied widely across the season. Transmission of O. vicarius between host social groups follows broadly predictable seasonal patterns, but there is nevertheless temporal and spatial heterogeneity in bug transmission. By understanding how long-distance movement by this vector varies in time and space, we can better predict where and when BCRV epizootics may occur.

Infestation by pyrethroids resistant bed bugs in the suburb of Paris, France

Directory of Open Access Journals (Sweden)

Durand R.

2012-11-01

Full Text Available Bed bugs are hematophagous insects responsible for a re-emerging and challenging indoor pest in many countries. Bed bugs infestations may have health consequences including nuisance biting, cutaneous and systemic reactions. This resurgence can probably be attributed to factors such as increased international travel and development of resistance against insecticides. Resistance against pyrethroids has been reported several times from the USA and rarely in Europe. In France, very few data on bed bugs are available. The present study aimed to assess the infestation by bed bugs of a complex of two high-rise apartment buildings in the suburb of Paris and to evaluate their susceptibility to pyrethroid insecticides. We inspected for bed bugs 192 out of 198 apartments units (97% and interviewed their residents. 76 (39.6% apartments were infested. Among the 97 residents living in infested apartments, 53 (54.6% reported bed bug bites. A total of 564 bed bugs were collected in the infested units. Bioassays showed that 54 out of 143 bed bugs were resistant to pyrethroids (37.8%; 95% confidence interval: 29.9-45.7%. DNA sequencing showed that all bed bugs tested (n = 124 had homozygous L925I kdr-like gene mutation. The level of pyrethroid resistance found indicates that this phenomenon was already established in the site and prompts the need to reevaluate the wide use of pyrethroids to control bed bugs.

Implementation of the e-Bug Project in Greece.

Science.gov (United States)

Gennimata, Dimitra; Merakou, Kyriakoula; Barbouni, Anastasia; Kremastinou, Jenny

2011-06-01

The e-Bug pack and web site educational material has been translated and adapted to the Greek language and educational background, and implemented throughout Greece as a supplementary educational resource in elementary and junior high schools. Elementary and junior high school teachers in Greece have actively participated in the development of the e-Bug educational resource and supported the implementation of all e-Bug activities. Dissemination to all key national stakeholders has been undertaken, and endorsement has been obtained from educational and medical associations, societies and institutions. Independent evaluation has been carried out, as part of dissertation thesis projects, for postgraduate studies. The e-Bug educational resource provides all the essentials for the dissemination of good health behaviours in hygiene, monitoring the spread of infection and the prudent use of antibiotics, to the youth of this country. Its contribution is expected to be evident in the next adult generation.

Bed Bug Myths

Science.gov (United States)

Learn the truth about bed bugs, such as how easy they are to see with the naked eye, their preferred habitat, whether they transmit diseases, their public health effects, and whether pesticides are the best way to deal with an infestation.

Learning from Mistakes --- A Comprehensive Study on Real World Concurrency Bug Characteristics

International Nuclear Information System (INIS)

Shan Lu; Soyeon Park; Eunsoo Seo; Yuanyuan Zhou

2008-01-01

The reality of multi-core hardware has made concurrent programs pervasive. Unfortunately, writing correct concurrent programs is difficult. Addressing this challenge requires advances in multiple directions, including concurrency bug detection, concurrent program testing, concurrent programming model design, etc. Designing effective techniques in all these directions will significantly benefit from a deep understanding of real world concurrency bug characteristics. This paper provides the first (to the best of our knowledge) comprehensive real world concurrency bug characteristic study. Specifically, we have carefully examined concurrency bug patterns, manifestation, and fix strategies of 105 randomly selected real world concurrency bugs from 4 representative server and client opensource applications (MySQL, Apache, Mozilla and OpenOffice). Our study reveals several interesting findings and provides useful guidance for concurrency bug detection, testing, and concurrent programming language design. Some of our findings are as follows: (1) Around one third of the examined non-deadlock concurrency bugs are caused by violation to programmers order intentions, which may not be easily expressed via synchronization primitives like locks and transactional memories; (2) Around 34% of the examined non-deadlock concurrency bugs involve multiple variables, which are not well addressed by existing bug detection tools; (3) About 92% of the examined concurrency bugs can be reliably triggered by enforcing certain orders among no more than 4 memory accesses. This indicates that testing concurrent programs can target at exploring possible orders among every small groups of memory accesses, instead of among all memory accesses; (4) About 73% of the examined non-deadlock concurrency bugs were not fixed by simply adding or changing locks, and many of the fixes were not correct at the first try, indicating the difficulty of reasoning concurrent execution by programmers.

Four bugs on a rectangle

KAUST Repository

Chapman, S. J.

2010-11-10

The idealized mathematical problem of four bugs in cyclic pursuit starting from a 2-by-1 rectangle is considered, and asymptotic formulas are derived to describe the motion. In contrast to the famous case of four bugs on a square, here the trajectories quickly freeze to essentially one dimension. After the first rotation about the centre point, the scale of the configuration has shrunk by a factor of 10427907250, and this number is then exponentiated four more times with each successive cycle. Relations to Knuth\\'s double-arrow notation and level-index arithmetic are discussed. This journal is © 2011 The Royal Society.

BUGS at work : a bicycle user group guide

Energy Technology Data Exchange (ETDEWEB)

NONE

2002-07-01

This guide provides practical advice to cyclists on how start a Bicycle User Group (BUG) at their workplace. It offers tools to encourage employers to be proactive in improving facilities in support of cycle commuting. Several BUGs across Canada have worked towards getting better bicycle parking, lockers and shower facilities at their workplace. Other incentives include policies such as flexible work hours for cyclist commuters; casual dress on Friday; reimbursement for the subsidized cost of free parking provided by employers; and, use of a company car if needed for company business during the work day. The advantages to employers include: reduced health care costs because cyclists are physically fit; decreased absenteeism; increased productivity; reduced parking costs; lower company transportation bills; and, a greener corporate image. BUGs provide cycling information ranging from cycling maps to pamphlets and they raise cycle awareness. This guide includes cycling survey samples and examples of successful BUG activities across Canada. refs., tabs., figs.

Quantum dot-induced viral capsid assembling in dissociation buffer

Directory of Open Access Journals (Sweden)

Gao D

2013-06-01

Full Text Available Ding Gao,1,2 Zhi-Ping Zhang,1 Feng Li,3 Dong Men,1 Jiao-Yu Deng,1 Hong-Ping Wei,1 Xian-En Zhang,1 Zong-Qiang Cui1 1State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 2Graduate University of Chinese Academy of Sciences, Beijing, 3Division of Nanobiomedicine and i-Lab, Suzhou Institute of Nano-Tech and Nano-Bionics, Chinese Academy of Sciences, Suzhou, People's Republic of China Abstract: Viruses encapsulating inorganic nanoparticles are a novel type of nanostructure with applications in biomedicine and biosensors. However, the encapsulation and assembly mechanisms of these hybridized virus-based nanoparticles (VNPs are still unknown. In this article, it was found that quantum dots (QDs can induce simian virus 40 (SV40 capsid assembly in dissociation buffer, where viral capsids should be disassembled. The analysis of the transmission electron microscope, dynamic light scattering, sucrose density gradient centrifugation, and cryo-electron microscopy single particle reconstruction experimental results showed that the SV40 major capsid protein 1 (VP1 can be assembled into ≈25 nm capsids in the dissociation buffer when QDs are present and that the QDs are encapsulated in the SV40 capsids. Moreover, it was determined that there is a strong affinity between QDs and the SV40 VP1 proteins (KD = 2.19E-10 M, which should play an important role in QD encapsulation in the SV40 viral capsids. This study provides a new understanding of the assembly mechanism of SV40 virus-based nanoparticles with QDs, which may help in the design and construction of other similar virus-based nanoparticles. Keywords: quantum dots, simian virus 40, self-assembly, encapsulation, virus-based nanoparticles

Human rhinovirus capsid dynamics is controlled by canyon flexibility

International Nuclear Information System (INIS)

Reisdorph, Nichole; Thomas, John J.; Katpally, Umesh; Chase, Elaine; Harris, Ken; Siuzdak, Gary; Smith, Thomas J.

2003-01-01

Quantitative enzyme accessibility experiments using nano liquid chromatography electrospray mass spectrometry combined with limited proteolysis and isotope-labeling was used to examine the dynamic nature of the human rhinovirus (HRV) capsid in the presence of three antiviral compounds, a neutralizing Fab, and drug binding cavity mutations. Using these methods, it was found that the antivirals WIN 52084 and picovir (pleconaril) stabilized the capsid, while dansylaziridine caused destabilization. Site-directed mutations in the drug-binding cavity were found to stabilize the HRV14 capsid against proteolytic digestion in a manner similar to WIN 52084 and pleconaril. Antibodies that bind to the NIm-IA antigenic site and penetrate the canyon were also observed to protect the virion against proteolytic cleavage. These results demonstrate that quantifying the effects of antiviral ligands on protein 'breathing' can be used to compare their mode of action and efficacy. In this case, it is apparent that hydrophobic antiviral agents, antibodies, or mutations in the canyon region block viral breathing. Therefore, these studies demonstrate that mobility in the canyon region is a major determinant in capsid breathing

Investigating the thermal dissociation of viral capsid by lattice model

Science.gov (United States)

Chen, Jingzhi; Chevreuil, Maelenn; Combet, Sophie; Lansac, Yves; Tresset, Guillaume

2017-11-01

The dissociation of icosahedral viral capsids was investigated by a homogeneous and a heterogeneous lattice model. In thermal dissociation experiments with cowpea chlorotic mottle virus and probed by small-angle neutron scattering, we observed a slight shrinkage of viral capsids, which can be related to the strengthening of the hydrophobic interaction between subunits at increasing temperature. By considering the temperature dependence of hydrophobic interaction in the homogeneous lattice model, we were able to give a better estimate of the effective charge. In the heterogeneous lattice model, two sets of lattice sites represented different capsid subunits with asymmetric interaction strengths. In that case, the dissociation of capsids was found to shift from a sharp one-step transition to a gradual two-step transition by weakening the hydrophobic interaction between AB and CC subunits. We anticipate that such lattice models will shed further light on the statistical mechanics underlying virus assembly and disassembly.

The chemical volatiles (semiochemicals) produced by neo tropical stink bugs (Hemiptera: Pentatomidae)

Energy Technology Data Exchange (ETDEWEB)

Moraes, Maria C.B.; Pareja, Martin; Laumann, Raul A.; Borges, Miguel [EMBRAPA Recursos Geneticos e Biotecnologia, Brasilia, DF (Brazil). Nucleo Tematico Controle Biologico

2008-09-15

In recent years the growing concern about environmental changes and how we are using the natural resources have triggered a search for natural products as alternatives to synthetic pesticides. The stink bugs produce a wide variety of chemical compounds (semiochemicals) that show potential to manage these insects. The stink bugs Chinavia impicticornis (Stal), C. ubica (Rolston), Dichelops melacanthus (Dallas), Euschistus heros (F.), Piezodorus guildinii (Westwood), Thyanta perditor (Westwood) and Tibraca limbativentris (Stal) had their blends of defensive compounds evaluated both qualitative and quantitatively. The main compounds identified on the glands of Brazilian stink bugs are: 2-alkenals, mainly the E isomer; saturated aliphatic hydrocarbons; and 4 oxo-(E)-2-alkenals. The first sex attractant determined from a stink bug was obtained from Nezara viridula L., and consists on a mix of two isomers cis - and trans bisabolene-epoxides. Later the soybean stink bug E. heros was also studied and its sex attractant was identified as three esters methyl: 2,6,10-trimethyl decanoate, methyl 2,6,10-trimethyl dodecanoate, and methyl E2, Z4-decadienoate. Recently, three new Brazilian sting bugs were studied and had their sex attractant elucidated. Males of T. perditor produce the ester, methyl 2E, 4Z, 6Z-decatrienoate. Whereas, the stink bug, P. guildinii has as sexual pheromone, the sesquiterpene beta-sesqui phellandrene, and the stink bug T. limbativentris produces as sex attractant the zingiberenol. In this review we discuss the advances obtained on the behaviour and identification of sex and defensive compound of stink bugs from Brazilian crops and the application of this knowledge to manage the stink bugs. (author)

The chemical volatiles (semiochemicals) produced by neo tropical stink bugs (Hemiptera: Pentatomidae)

International Nuclear Information System (INIS)

Moraes, Maria C.B.; Pareja, Martin; Laumann, Raul A.; Borges, Miguel

2008-01-01

In recent years the growing concern about environmental changes and how we are using the natural resources have triggered a search for natural products as alternatives to synthetic pesticides. The stink bugs produce a wide variety of chemical compounds (semiochemicals) that show potential to manage these insects. The stink bugs Chinavia impicticornis (Stal), C. ubica (Rolston), Dichelops melacanthus (Dallas), Euschistus heros (F.), Piezodorus guildinii (Westwood), Thyanta perditor (Westwood) and Tibraca limbativentris (Stal) had their blends of defensive compounds evaluated both qualitative and quantitatively. The main compounds identified on the glands of Brazilian stink bugs are: 2-alkenals, mainly the E isomer; saturated aliphatic hydrocarbons; and 4 oxo-(E)-2-alkenals. The first sex attractant determined from a stink bug was obtained from Nezara viridula L., and consists on a mix of two isomers cis - and trans bisabolene-epoxides. Later the soybean stink bug E. heros was also studied and its sex attractant was identified as three esters methyl: 2,6,10-trimethyl decanoate, methyl 2,6,10-trimethyl dodecanoate, and methyl E2, Z4-decadienoate. Recently, three new Brazilian sting bugs were studied and had their sex attractant elucidated. Males of T. perditor produce the ester, methyl 2E, 4Z, 6Z-decatrienoate. Whereas, the stink bug, P. guildinii has as sexual pheromone, the sesquiterpene beta-sesqui phellandrene, and the stink bug T. limbativentris produces as sex attractant the zingiberenol. In this review we discuss the advances obtained on the behaviour and identification of sex and defensive compound of stink bugs from Brazilian crops and the application of this knowledge to manage the stink bugs. (author)

Revised Mimivirus major capsid protein sequence reveals intron-containing gene structure and extra domain

Directory of Open Access Journals (Sweden)

Suzan-Monti Marie

2009-05-01

Full Text Available Abstract Background Acanthamoebae polyphaga Mimivirus (APM is the largest known dsDNA virus. The viral particle has a nearly icosahedral structure with an internal capsid shell surrounded with a dense layer of fibrils. A Capsid protein sequence, D13L, was deduced from the APM L425 coding gene and was shown to be the most abundant protein found within the viral particle. However this protein remained poorly characterised until now. A revised protein sequence deposited in a database suggested an additional N-terminal stretch of 142 amino acids missing from the original deduced sequence. This result led us to investigate the L425 gene structure and the biochemical properties of the complete APM major Capsid protein. Results This study describes the full length 3430 bp Capsid coding gene and characterises the 593 amino acids long corresponding Capsid protein 1. The recombinant full length protein allowed the production of a specific monoclonal antibody able to detect the Capsid protein 1 within the viral particle. This protein appeared to be post-translationnally modified by glycosylation and phosphorylation. We proposed a secondary structure prediction of APM Capsid protein 1 compared to the Capsid protein structure of Paramecium Bursaria Chlorella Virus 1, another member of the Nucleo-Cytoplasmic Large DNA virus family. Conclusion The characterisation of the full length L425 Capsid coding gene of Acanthamoebae polyphaga Mimivirus provides new insights into the structure of the main Capsid protein. The production of a full length recombinant protein will be useful for further structural studies.

Bug bites and stings: When to see a dermatologist

Medline Plus

Full Text Available ... var c = 0; c Tips to prevent and treat bug bites Although most bug bites are harmless, some can spread dangerous diseases like Zika virus, dengue, Lyme disease, and malaria. Particularly if you’re ...

Automated Bug Assignment: Ensemble-based Machine Learning in Large Scale Industrial Contexts

OpenAIRE

Jonsson, Leif; Borg, Markus; Broman, David; Sandahl, Kristian; Eldh, Sigrid; Runeson, Per

2016-01-01

Bug report assignment is an important part of software maintenance. In particular, incorrect assignments of bug reports to development teams can be very expensive in large software development projects. Several studies propose automating bug assignment techniques using machine learning in open source software contexts, but no study exists for large-scale proprietary projects in industry. The goal of this study is to evaluate automated bug assignment techniques that are based on machine learni...

Modified bug-1 algorithm based strategy for obstacle avoidance in multi robot system

Directory of Open Access Journals (Sweden)

Kandathil Jom J.

2018-01-01

Full Text Available One of the primary ability of an intelligent mobile robot system is obstacle avoidance. BUG algorithms are classic examples of the algorithms used for achieving obstacle avoidance. Unlike many other planning algorithms based on global knowledge, BUG algorithms assume only local knowledge of the environment and a global goal. Among the variations of the BUG algorithms that prevail, BUG-0, BUG-1 and BUG-2 are the more prominent versions. The exhaustive search algorithm present in BUG-1 makes it more reliable and safer for practical applications. Overall, this provides a more predictable and dependable performance. Hence, the essential focus in this paper is on implementing the BUG-1 algorithm across a group of robots to move them from a start location to a target location. The results are compared with the results from BUG-1 algorithm implemented on a single robot. The strategy developed in this work reduces the time involved in moving the robots from starting location to the target location. Further, the paper shows that the total distance covered by each robot in a multi robot-system is always lesser than or equal to that travelled by a single robot executing the same problem.

Bed Bug Information Clearinghouse

Science.gov (United States)

Its purpose is to help states, communities, and consumers in efforts to prevent and control bed bug infestations. Currently includes only reviewed material from federal/state/local government agencies, extension services, and universities.

AAV8 capsid variable regions at the two-fold symmetry axis contribute to high liver transduction by mediating nuclear entry and capsid uncoating

International Nuclear Information System (INIS)

Tenney, Rebeca M.; Bell, Christie L.; Wilson, James M.

2014-01-01

Adeno-associated virus serotype 8 (AAV8) is a promising vector for liver-directed gene therapy. Although efficient uncoating of viral capsids has been implicated in AAV8's robust liver transduction, much about the biology of AAV8 hepatotropism remains unclear. Our study investigated the structural basis of AAV8 liver transduction efficiency by constructing chimeric vector capsids containing sequences derived from AAV8 and AAV2 – a highly homologous yet poorly hepatotropic serotype. Engineered vectors containing capsid variable regions (VR) VII and IX from AAV8 in an AAV2 backbone mediated near AAV8-like transduction in mouse liver, with higher numbers of chimeric genomes detected in whole liver cells and isolated nuclei. Interestingly, chimeric capsids within liver nuclei also uncoated similarly to AAV8 by 6 weeks after administration, in contrast with AAV2, of which a significantly smaller proportion were uncoated. This study links specific AAV capsid regions to the transduction ability of a clinically relevant AAV serotype. - Highlights: • We construct chimeric vectors to identify determinants of AAV8 liver transduction. • An AAV2-based vector with 17 AAV8 residues exhibited high liver transduction in mice. • This vector also surpassed AAV2 in cell entry, nuclear entry and onset of expression. • Most chimeric vector particles were uncoated at 6 weeks, like AAV8 and unlike AAV2. • Chimera retained heparin binding and was antigenically distinct from AAV2 and AAV8

AAV8 capsid variable regions at the two-fold symmetry axis contribute to high liver transduction by mediating nuclear entry and capsid uncoating

Energy Technology Data Exchange (ETDEWEB)

Tenney, Rebeca M.; Bell, Christie L.; Wilson, James M., E-mail: wilsonjm@mail.med.upenn.edu

2014-04-15

Adeno-associated virus serotype 8 (AAV8) is a promising vector for liver-directed gene therapy. Although efficient uncoating of viral capsids has been implicated in AAV8's robust liver transduction, much about the biology of AAV8 hepatotropism remains unclear. Our study investigated the structural basis of AAV8 liver transduction efficiency by constructing chimeric vector capsids containing sequences derived from AAV8 and AAV2 – a highly homologous yet poorly hepatotropic serotype. Engineered vectors containing capsid variable regions (VR) VII and IX from AAV8 in an AAV2 backbone mediated near AAV8-like transduction in mouse liver, with higher numbers of chimeric genomes detected in whole liver cells and isolated nuclei. Interestingly, chimeric capsids within liver nuclei also uncoated similarly to AAV8 by 6 weeks after administration, in contrast with AAV2, of which a significantly smaller proportion were uncoated. This study links specific AAV capsid regions to the transduction ability of a clinically relevant AAV serotype. - Highlights: • We construct chimeric vectors to identify determinants of AAV8 liver transduction. • An AAV2-based vector with 17 AAV8 residues exhibited high liver transduction in mice. • This vector also surpassed AAV2 in cell entry, nuclear entry and onset of expression. • Most chimeric vector particles were uncoated at 6 weeks, like AAV8 and unlike AAV2. • Chimera retained heparin binding and was antigenically distinct from AAV2 and AAV8.

Biochemical characterization of digestive amylase of wheat bug ...

African Journals Online (AJOL)

STORAGESEVER

2009-08-04

Aug 4, 2009 ... ... Boyd et al., 2002). Wheat bug also feeds on different stage of developing grains. ... the grain during feeding, Wheat bug enzymes degrade gluten proteins and cause rapid relaxation of dough resulting in the production of .... A blank without substrate but with α-amylase extract and a control containing no ...

Zero bugs and program faster

CERN Document Server

Thompson, Kate

2015-01-01

A book about programming, improving skill, and avoiding mistakes. The author spent two years researching every bug avoidance technique she could find. This book contains the best of them. If you want to program faster, with fewer bugs, and write more secure code, buy this book! "This is the best book I have ever read." - Anonymous reviewer "Four score and seven years ago this book helped me debug my server code." -Abraham Lincoln "Would my Javascript have memory leaks without this book? Would fishes fly without water?" -Socrates "This book is the greatest victory since the Spanish Armada, and the best about programming." -Queen Elizabeth

WinBUGSio: A SAS Macro for the Remote Execution of WinBUGS

Directory of Open Access Journals (Sweden)

Michael K. Smith

2007-09-01

Full Text Available This is a macro which facilitates remote execution of WinBUGS from within SAS. The macro pre-processes data for WinBUGS, writes the WinBUGS batch-script, executes this script and reads in output statistics from the WinBUGS log-file back into SAS native format. The user specifies the input and output file names and directory path as well as the statistics to be monitored in WinBUGS. The code works best for a model that has already been set up and checked for convergence diagnostics within WinBUGS. An obvious extension of the use of this macro is for running simulations where the input and output files all have the same name but all that differs between simulation iterations is the input dataset. The functionality and syntax of the macro call are described in this paper and illustrated using a simple linear regression model.

ATP-Driven Contraction of Phage T3 Capsids with DNA Incompletely Packaged In Vivo

Directory of Open Access Journals (Sweden)

Philip Serwer

2017-05-01

Full Text Available Adenosine triphosphate (ATP cleavage powers packaging of a double-stranded DNA (dsDNA molecule in a pre-assembled capsid of phages that include T3. Several observations constitute a challenge to the conventional view that the shell of the capsid is energetically inert during packaging. Here, we test this challenge by analyzing the in vitro effects of ATP on the shells of capsids generated by DNA packaging in vivo. These capsids retain incompletely packaged DNA (ipDNA and are called ipDNA-capsids; the ipDNA-capsids are assumed to be products of premature genome maturation-cleavage. They were isolated via preparative Nycodenz buoyant density centrifugation. For some ipDNA-capsids, Nycodenz impermeability increases hydration and generates density so low that shell hyper-expansion must exist to accommodate associated water. Electron microscopy (EM confirmed hyper-expansion and low permeability and revealed that 3.0 mM magnesium ATP (physiological concentration causes contraction of hyper-expanded, lowpermeability ipDNA-capsids to less than mature size; 5.0 mM magnesium ATP (border of supraphysiological concentration or more disrupts them. Additionally, excess sodium ADP reverses 3.0 mM magnesium ATP-induced contraction and re-generates hyper-expansion. The Nycodenz impermeability implies assembly perfection that suggests selection for function in DNA packaging. These findings support the above challenge and can be explained via the assumption that T3 DNA packaging includes a back-up cycle of ATP-driven capsid contraction and hyper-expansion.

A molecular breadboard: Removal and replacement of subunits in a hepatitis B virus capsid.

Science.gov (United States)

Lee, Lye Siang; Brunk, Nicholas; Haywood, Daniel G; Keifer, David; Pierson, Elizabeth; Kondylis, Panagiotis; Wang, Joseph Che-Yen; Jacobson, Stephen C; Jarrold, Martin F; Zlotnick, Adam

2017-11-01

Hepatitis B virus (HBV) core protein is a model system for studying assembly and disassembly of icosahedral structures. Controlling disassembly will allow re-engineering the 120 subunit HBV capsid, making it a molecular breadboard. We examined removal of subunits from partially crosslinked capsids to form stable incomplete particles. To characterize incomplete capsids, we used two single molecule techniques, resistive-pulse sensing and charge detection mass spectrometry. We expected to find a binomial distribution of capsid fragments. Instead, we found a preponderance of 3 MDa complexes (90 subunits) and no fragments smaller than 3 MDa. We also found 90-mers in the disassembly of uncrosslinked HBV capsids. 90-mers seem to be a common pause point in disassembly reactions. Partly explaining this result, graph theory simulations have showed a threshold for capsid stability between 80 and 90 subunits. To test a molecular breadboard concept, we showed that missing subunits could be refilled resulting in chimeric, 120 subunit particles. This result may be a means of assembling unique capsids with functional decorations. © 2017 The Protein Society.

Developing e-Bug web games to teach microbiology.

Science.gov (United States)

Farrell, David; Kostkova, Patty; Lazareck, Lisa; Weerasinghe, Dasun; Weinberg, Julius; Lecky, Donna M; Adriaenssens, Niels; Koprivová Herotová, Tereza; Holt, Jette; Touboul, Pia; Merakou, Kyriakoula; Koncan, Raffaella; Olczak-Pienkowska, Anna; Avô, António Brito; Campos, José; McNulty, Cliodna A M

2011-06-01

As a complement to the e-Bug teaching pack, two e-Bug games were developed to provide content that aimed to entertain as well as to educate. A set of agreed learning outcomes (LOs) were provided by the scientific partners of the e-Bug Project and the games were developed using user-centred design techniques (the needs, wants and limitations of the potential game players were assessed at each stage of the design process). The e-Bug games were designed for two age groups: Junior (9-12 year olds); and Senior (13-15 year olds). A study using focus groups was done to gain an understanding as to the types of games enjoyed by the target users. According to the preliminary study, the Junior Game was developed as a platform game and the Senior Game was developed as a story-based detective game. The Junior Game consists of five levels, each associated with a set of LOs. Similarly, the Senior Game consists of four missions, each comprising five stages using problem-based learning techniques and LOs. In this paper, the process of development for each game is described in detail and an illustration is provided of how each game level or mission addresses the target LOs. Development of the games used feedback acquired from children in four schools across the UK (Glasgow, London and two in Gloucester). The children were selected according to their willingness to participate. European Partners of the e-Bug Project also provided further support, translation and requests for modifications. The knowledge gained of LOs and further evaluation of the games is continuing, and preliminary results are in press. The final versions of the games, translated into 11 European languages, are available online via www.e-bug.eu.

Bed Bug Epidemic: A Challenge to Public Health

Science.gov (United States)

Ratnapradipa, Dhitinut; Ritzel, Dale O.; Haramis, Linn D.; Bliss, Kadi R.

2011-01-01

In recent years, reported cases of bed bug infestations in the U.S. and throughout the world have escalated dramatically, posing a global public health problem. Although bed bugs are not known to transmit disease to humans, they pose both direct and indirect public health challenges in terms of health effects, treatment, cost, and resource…

Nanobodies targeting norovirus capsid reveal functional epitopes and potential mechanisms of neutralization.

Directory of Open Access Journals (Sweden)

Anna D Koromyslova

2017-11-01

Full Text Available Norovirus is the leading cause of gastroenteritis worldwide. Despite recent developments in norovirus propagation in cell culture, these viruses are still challenging to grow routinely. Moreover, little is known on how norovirus infects the host cells, except that histo-blood group antigens (HBGAs are important binding factors for infection and cell entry. Antibodies that bind at the HBGA pocket and block attachment to HBGAs are believed to neutralize the virus. However, additional neutralization epitopes elsewhere on the capsid likely exist and impeding the intrinsic structural dynamics of the capsid could be equally important. In the current study, we investigated a panel of Nanobodies in order to probe functional epitopes that could trigger capsid rearrangement and/ or interfere with HBGA binding interactions. The precise binding sites of six Nanobodies (Nano-4, Nano-14, Nano-26, Nano-27, Nano-32, and Nano-42 were identified using X-ray crystallography. We showed that these Nanobodies bound on the top, side, and bottom of the norovirus protruding domain. The impact of Nanobody binding on norovirus capsid morphology was analyzed using electron microscopy and dynamic light scattering. We discovered that distinct Nanobody epitopes were associated with varied changes in particle structural integrity and assembly. Interestingly, certain Nanobody-induced capsid morphological changes lead to the capsid protein degradation and viral RNA exposure. Moreover, Nanobodies employed multiple inhibition mechanisms to prevent norovirus attachment to HBGAs, which included steric obstruction (Nano-14, allosteric interference (Nano-32, and violation of normal capsid morphology (Nano-26 and Nano-85. Finally, we showed that two Nanobodies (Nano-26 and Nano-85 not only compromised capsid integrity and inhibited VLPs attachment to HBGAs, but also recognized a broad panel of norovirus genotypes with high affinities. Consequently, Nano-26 and Nano-85 have a great

Nanobodies targeting norovirus capsid reveal functional epitopes and potential mechanisms of neutralization

Science.gov (United States)

2017-01-01

Norovirus is the leading cause of gastroenteritis worldwide. Despite recent developments in norovirus propagation in cell culture, these viruses are still challenging to grow routinely. Moreover, little is known on how norovirus infects the host cells, except that histo-blood group antigens (HBGAs) are important binding factors for infection and cell entry. Antibodies that bind at the HBGA pocket and block attachment to HBGAs are believed to neutralize the virus. However, additional neutralization epitopes elsewhere on the capsid likely exist and impeding the intrinsic structural dynamics of the capsid could be equally important. In the current study, we investigated a panel of Nanobodies in order to probe functional epitopes that could trigger capsid rearrangement and/ or interfere with HBGA binding interactions. The precise binding sites of six Nanobodies (Nano-4, Nano-14, Nano-26, Nano-27, Nano-32, and Nano-42) were identified using X-ray crystallography. We showed that these Nanobodies bound on the top, side, and bottom of the norovirus protruding domain. The impact of Nanobody binding on norovirus capsid morphology was analyzed using electron microscopy and dynamic light scattering. We discovered that distinct Nanobody epitopes were associated with varied changes in particle structural integrity and assembly. Interestingly, certain Nanobody-induced capsid morphological changes lead to the capsid protein degradation and viral RNA exposure. Moreover, Nanobodies employed multiple inhibition mechanisms to prevent norovirus attachment to HBGAs, which included steric obstruction (Nano-14), allosteric interference (Nano-32), and violation of normal capsid morphology (Nano-26 and Nano-85). Finally, we showed that two Nanobodies (Nano-26 and Nano-85) not only compromised capsid integrity and inhibited VLPs attachment to HBGAs, but also recognized a broad panel of norovirus genotypes with high affinities. Consequently, Nano-26 and Nano-85 have a great potential to

L2, the minor capsid protein of papillomavirus

Energy Technology Data Exchange (ETDEWEB)

Wang, Joshua W. [Department of Pathology, The Johns Hopkins University, Baltimore, MD 21287 (United States); Roden, Richard B.S., E-mail: roden@jhmi.edu [Department of Pathology, The Johns Hopkins University, Baltimore, MD 21287 (United States); Department of Oncology, The Johns Hopkins University, Baltimore, MD 21287 (United States); Department of Gynecology and Obstetrics, The Johns Hopkins University, Baltimore, MD 21287 (United States)

2013-10-15

The capsid protein L2 plays major roles in both papillomavirus assembly and the infectious process. While L1 forms the majority of the capsid and can self-assemble into empty virus-like particles (VLPs), L2 is a minor capsid component and lacks the capacity to form VLPs. However, L2 co-assembles with L1 into VLPs, enhancing their assembly. L2 also facilitates encapsidation of the ∼8 kbp circular and nucleosome-bound viral genome during assembly of the non-enveloped T=7d virions in the nucleus of terminally differentiated epithelial cells, although, like L1, L2 is not detectably expressed in infected basal cells. With respect to infection, L2 is not required for particles to bind to and enter cells. However L2 must be cleaved by furin for endosome escape. L2 then travels with the viral genome to the nucleus, wherein it accumulates at ND-10 domains. Here, we provide an overview of the biology of L2. - Highlights: • L2 is the minor antigen of the non-enveloped T=7d icosahedral Papillomavirus capsid. • L2 is a nuclear protein that can traffic to ND-10 and facilitate genome encapsidation. • L2 is critical for infection and must be cleaved by furin. • L2 is a broadly protective vaccine antigen recognized by neutralizing antibodies.

L2, the minor capsid protein of papillomavirus

International Nuclear Information System (INIS)

Wang, Joshua W.; Roden, Richard B.S.

2013-01-01

The capsid protein L2 plays major roles in both papillomavirus assembly and the infectious process. While L1 forms the majority of the capsid and can self-assemble into empty virus-like particles (VLPs), L2 is a minor capsid component and lacks the capacity to form VLPs. However, L2 co-assembles with L1 into VLPs, enhancing their assembly. L2 also facilitates encapsidation of the ∼8 kbp circular and nucleosome-bound viral genome during assembly of the non-enveloped T=7d virions in the nucleus of terminally differentiated epithelial cells, although, like L1, L2 is not detectably expressed in infected basal cells. With respect to infection, L2 is not required for particles to bind to and enter cells. However L2 must be cleaved by furin for endosome escape. L2 then travels with the viral genome to the nucleus, wherein it accumulates at ND-10 domains. Here, we provide an overview of the biology of L2. - Highlights: • L2 is the minor antigen of the non-enveloped T=7d icosahedral Papillomavirus capsid. • L2 is a nuclear protein that can traffic to ND-10 and facilitate genome encapsidation. • L2 is critical for infection and must be cleaved by furin. • L2 is a broadly protective vaccine antigen recognized by neutralizing antibodies

Catching the Bug: How Virtual Coaching Improves Teaching

Science.gov (United States)

Allen, Megan

2014-01-01

In this article the author describes virtual coaching and why it is so effective. The following six points of virtual coaching are explained: (1) Also known as bug-in-ear coaching, virtual coaching is not new; (2) Virtual coaching can save money and time; (3) Bug-in-ear coaching increases the frequency of observations for novice teachers; (4) It…

High Relaxivity Gadolinium Hydroxypyridonate-Viral Capsid Conjugates: Nano-sized MRI Contrast Agents

Energy Technology Data Exchange (ETDEWEB)

Meux, Susan C.; Datta, Ankona; Hooker, Jacob M.; Botta, Mauro; Francis, Matthew B.; Aime, Silvio; Raymond, Kenneth N.

2007-08-29

High relaxivity macromolecular contrast agents based on the conjugation of gadolinium chelates to the interior and exterior surfaces of MS2 viral capsids are assessed. The proton nuclear magnetic relaxation dispersion (NMRD) profiles of the conjugates show up to a five-fold increase in relaxivity, leading to a peak relaxivity (per Gd{sup 3+} ion) of 41.6 mM{sup -1}s{sup -1} at 30 MHz for the internally modified capsids. Modification of the exterior was achieved through conjugation to flexible lysines, while internal modification was accomplished by conjugation to relatively rigid tyrosines. Higher relaxivities were obtained for the internally modified capsids, showing that (1) there is facile diffusion of water to the interior of capsids and (2) the rigidity of the linker attaching the complex to the macromolecule is important for obtaining high relaxivity enhancements. The viral capsid conjugated gadolinium hydroxypyridonate complexes appear to possess two inner-sphere water molecules (q = 2) and the NMRD fittings highlight the differences in the local motion for the internal ({tau}{sub RI} = 440 ps) and external ({tau}{sub RI} = 310 ps) conjugates. These results indicate that there are significant advantages of using the internal surface of the capsids for contrast agent attachment, leaving the exterior surface available for the installation of tissue targeting groups.

The Mammalian Cell Cycle Regulates Parvovirus Nuclear Capsid Assembly

Science.gov (United States)

Riolobos, Laura; Domínguez, Carlos; Kann, Michael; Almendral, José M.

2015-01-01

It is unknown whether the mammalian cell cycle could impact the assembly of viruses maturing in the nucleus. We addressed this question using MVM, a reference member of the icosahedral ssDNA nuclear parvoviruses, which requires cell proliferation to infect by mechanisms partly understood. Constitutively expressed MVM capsid subunits (VPs) accumulated in the cytoplasm of mouse and human fibroblasts synchronized at G0, G1, and G1/S transition. Upon arrest release, VPs translocated to the nucleus as cells entered S phase, at efficiencies relying on cell origin and arrest method, and immediately assembled into capsids. In synchronously infected cells, the consecutive virus life cycle steps (gene expression, proteins nuclear translocation, capsid assembly, genome replication and encapsidation) proceeded tightly coupled to cell cycle progression from G0/G1 through S into G2 phase. However, a DNA synthesis stress caused by thymidine irreversibly disrupted virus life cycle, as VPs became increasingly retained in the cytoplasm hours post-stress, forming empty capsids in mouse fibroblasts, thereby impairing encapsidation of the nuclear viral DNA replicative intermediates. Synchronously infected cells subjected to density-arrest signals while traversing early S phase also blocked VPs transport, resulting in a similar misplaced cytoplasmic capsid assembly in mouse fibroblasts. In contrast, thymidine and density arrest signals deregulating virus assembly neither perturbed nuclear translocation of the NS1 protein nor viral genome replication occurring under S/G2 cycle arrest. An underlying mechanism of cell cycle control was identified in the nuclear translocation of phosphorylated VPs trimeric assembly intermediates, which accessed a non-conserved route distinct from the importin α2/β1 and transportin pathways. The exquisite cell cycle-dependence of parvovirus nuclear capsid assembly conforms a novel paradigm of time and functional coupling between cellular and virus life

Posttreatment Feeding Affects Mortality of Bed Bugs (Hemiptera: Cimicidae) Exposed to Insecticides.

Science.gov (United States)

Singh, Narinderpal; Wang, Changlu; Cooper, Richard

2016-02-01

Insecticide sprays and dusts are used for controlling bed bugs, Cimex lectularius L. In natural environments, bed bugs have daily access to hosts after they are exposed to insecticides. The established laboratory insecticide bioassay protocols do not provide feeding after insecticide treatments, which can result in inflated mortality compared with what would be encountered in the field. We evaluated the effect of posttreatment feeding on mortality of bed bugs treated with different insecticides. None of the insecticides tested had a significant effect on the amount of blood consumed and percent feeding. The effect of posttreatment feeding on bed bug mortality varied among different insecticides. Feeding significantly reduced mortality in bed bugs exposed to deltamethrin spray, an essential oil mixture (Bed Bug Fix) spray, and diatomaceous earth dust. Feeding increased the mean survival time for bed bugs treated with chlorfenapyr spray and a spray containing an essential oil mixture (Ecoraider), but did not affect the final mortality. First instars hatched from eggs treated with chlorfenapyr liquid spray had reduced feeding compared with nymphs hatched from nontreated eggs. Those nymphs hatched from eggs treated with chlorfenapyr liquid spray and successfully fed had reduced mortality and a higher mean survival time than those without feeding. We conclude that the availability of a bloodmeal after insecticide exposure has a significant effect on bed bug mortality. Protocols for insecticide efficacy testing should consider offering a bloodmeal to the treated bed bugs within 1 to 3 d after treatment. © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Structure of the capsid of Kilham rat virus from small-angle neutron scattering

International Nuclear Information System (INIS)

Wobbe, C.R.; Mitra, S.; Ramakrishnan, V.

1984-01-01

The structure of empty capsids of Kilham rat virus, an autonomous parvovirus with icosahedral symmetry, was investigated by small-angle neutron scattering. From the forward scatter, the molecular weight was determined to be 4.0 x 10(6), and from the Guinier region, the radius of gyration was found to be 105 A in D2O and 104 A in H 2 O. On the basis of the capsid molecular weight and the molecular weights and relative abundances of the capsid proteins, the authors propose that the capsid has a triangulation number of 1. Extended scattering curves and mathematical modeling revealed that the capsid consists of two shells of protein, the inner shell extending from 58 to 91 A in D2O and from 50 to 91 A in H 2 O and containing 11% of the capsid scattering mass, and the outer shell extending to 121 A in H 2 O and D2O. The inner shell appears to have a higher content of basic amino acids than the outer shell, based on its lower scattering density in D2O than in H 2 O. The authors propose that all three capsid proteins contribute to the inner shell and that this basic region serves DNA binding and partial charge neutralization functions

Intracellular cargo delivery by virus capsid protein-based vehicles: From nano to micro.

Science.gov (United States)

Gao, Ding; Lin, Xiu-Ping; Zhang, Zhi-Ping; Li, Wei; Men, Dong; Zhang, Xian-En; Cui, Zong-Qiang

2016-02-01

Cellular delivery is an important concern for the efficiency of medicines and sensors for disease diagnoses and therapy. However, this task is quite challenging. Self-assembly virus capsid proteins might be developed as building blocks for multifunctional cellular delivery vehicles. In this work, we found that SV40 VP1 (Simian virus 40 major capsid protein) could function as a new cell-penetrating protein. The VP1 protein could carry foreign proteins into cells in a pentameric structure. A double color structure, with red QDs (Quantum dots) encapsulated by viral capsids fused with EGFP, was created for imaging cargo delivery and release from viral capsids. The viral capsids encapsulating QDs were further used for cellular delivery of micron-sized iron oxide particles (MPIOs). MPIOs were efficiently delivered into live cells and controlled by a magnetic field. Therefore, our study built virus-based cellular delivery systems for different sizes of cargos: protein molecules, nanoparticles, and micron-sized particles. Much research is being done to investigate methods for efficient and specific cellular delivery of drugs, proteins or genetic material. In this article, the authors describe their approach in using self-assembly virus capsid proteins SV40 VP1 (Simian virus 40 major capsid protein). The cell-penetrating behavior provided excellent cellular delivery and should give a new method for biomedical applications. Copyright © 2015 Elsevier Inc. All rights reserved.

Bed Bugs and Schools

Science.gov (United States)

Bed bugs have long been a pest – feeding on blood, causing itchy bites and generally irritating their human hosts. They are successful hitchhikers, and can move from an infested site to furniture, bedding, baggage, boxes, and clothing.

Essential oils as fumigants for bed bugs (Hemiptera: Cimicidae)

Science.gov (United States)

In Petri dish assays, fumigation of a pyrethroid-susceptible strain of bed bugs Cimex lectularius L. (Hemiptera: Cimicidae) with various essential oils resulted in mortality that approached or equaled 100%, after 5 days. However, when bed bugs were exposed to the same essential oils in sealed, comme...

Effects of Starvation on Deltamethrin Tolerance in Bed Bugs, Cimex lectularius L. (Hemiptera: Cimicidae

Directory of Open Access Journals (Sweden)

Zachary C. DeVries

2015-01-01

Full Text Available Bed bugs, Cimex lectularius L., are a major pest in the urban environment. Their presence often results in physical, psychological, and financial distress of homeowners and apartment dwellers. Although many insecticide bioassays have been performed on this pest, little attention has been paid to bed bug feeding status, which is closely linked to metabolism, molting, and mass. Therefore, we evaluated the toxicity of topically applied deltamethrin on insecticide susceptible adult male bed bugs fed 2 d, 9 d, and 21 d prior to testing. When toxicity was evaluated on a “per-bug” basis, there was no difference between 2 d [LD50 = 0.498 (0.316 − 0.692 ng·bug−1] and 9 d [LD50 = 0.572 (0.436 − 0.724 ng·bug−1] starved bugs, while 21 d starved bugs had a significantly lower LD50 [0.221 (0.075 − 0.386 ng·bug−1]. When toxicity was evaluated in terms of body mass, 9 d starved bugs had the highest LD50 values [0.138 (0.102 − 0.176 ng·mg−1], followed by 2 d starved bugs [0.095 (0.060 − 0.134 ng·mg−1], and then 21 d starved bugs [0.058 (0.019–0.102 ng·mg−1]; the LD50 values of 2 d and 9 d starved bugs were significantly different from 21 d starved bugs. These results indicate that feeding status plays an important role in the toxicity of deltamethrin. In addition, the lack of differences between 2 d and 9 d starved bugs indicate that the blood meal itself has little impact on tolerance, but rather it is some physiological change following feeding that confers increased tolerance to bed bugs.

Comparison of Three Bed Bug Management Strategies in a Low-Income Apartment Building

Directory of Open Access Journals (Sweden)

Changlu Wang

2012-04-01

Full Text Available Bed bug (Cimex lectularius L. infestations are currently controlled by a variety of non-chemical and chemical methods. There have been few studies on the comparative effectiveness of these control techniques. We evaluated three bed bug management strategies in an apartment building: (1 non-chemical methods only (n = 9; (2 insecticides only (n = 6; and (3 integrated pest management including both non-chemical methods and insecticides (n = 9. The apartments were one-bedroom units occupied by seniors or people with disabilities. Bed bug numbers in each apartment were determined by visual inspection and/or installing intercepting devices under bed and sofa legs. The median (min, max bed bug counts in the non-chemical methods only, insecticides only, and integrated pest management (IPM treatment were: 4 (1, 57, 19 (1, 250, and 14 (1, 219, respectively prior to the treatments. The apartments were retreated if found necessary during biweekly to monthly inspections. After 10 weeks, bed bugs were found to be eliminated from 67, 33, and 44% of the apartments in the three treatment groups, respectively. The final (after 10 weeks median (min, max bed bug counts in the non-chemical methods only, insecticides only, and IPM treatment were: 0 (0, 134, 11.5 (0, 58, and 1 (0, 38, respectively. There were no significant differences in the speed of bed bug count reduction or the final bed bug counts. Lack of resident cooperation partially contributed to the failure in eliminating bed bugs from some of the apartments. Results of this study suggest that non-chemical methods can effectively eliminate bed bugs in lightly infested apartments.

Prevalence of bed bug (Cimex lectularius in human settlement area of Bahnamir, Iran

Directory of Open Access Journals (Sweden)

Seyyed Farzad Motevalli Haghi

2014-09-01

Full Text Available Objective: To survey the factors concerning with bed bugs epidemiology among human settlement area of Bahnamir, Iran in 2012-2013. Methods: Bed bugs were collected from 500 households which lived in eight villages in suburban areas. After random cluster classification, we checked their houses to collect information on presence of bed bugs. When bed bugs infestation revealed, they were preserved in properly labeled specimen containing 70% alcohol. Bugs were transferred to the Faculty of Health and identified by stereomicroscopy. A questionnaire was filled by the owners. The recorded data were coded and analyzed statistically using the Chi-square test. Results: In the study area, 14 from 500 families were infested by bed bugs which revealed almost 2.8% of infestation in that area. Five infested villages had shown 62.50% infestation. From the total of 256 caught bugs, 56.45%, 31.25%, 8.59%, and 3.91% was separated from bedroom, setting room, kitchen and other parts of the infested houses, respectively. Regarding to 256 trapped bugs in the bedroom, 33.2% were immature (nymph and 66.8% were mature. Conclusions: The results illustrated the role of education and guidance of the owners towards improving personal hygiene and public health with using insecticides to control by professional persons, not using secondhand instrument in the home and washing sheets and blankets and drying on suitable hot drier in order to reduce the infestation rate of bed bug among human settlement.

Ecosystem-Based Incorporation of Nectar-Producing Plants for Stink Bug Parasitoids

Directory of Open Access Journals (Sweden)

Glynn Tillman

2017-06-01

Full Text Available Adult parasitoids of pest insects rely on floral resources for survival and reproduction, but can be food-deprived in intensively managed agricultural systems lacking these resources. Stink bugs are serious pests for crops in southwest Georgia. Provisioning nectar-producing plants for parasitoids of stink bugs potentially can enhance biocontrol of these pests. Knowledge of spatial and temporal availability and distribution of stink bugs in host plants is necessary for appropriate timing and placement of flowering plants in agroecosystems. Stink bugs move between closely associated host plants throughout the growing season in response to deteriorating suitability of their host plants. In peanut-cotton farmscapes, stink bugs develop in peanut, and subsequently the adults disperse into adjacent cotton. Parasitism of Nezara viridula (L. adults by Trichopoda pennipes (F. at the peanut-cotton interface was significantly higher in cotton with a strip of milkweed or buckwheat between the two crops than in cotton alone. Milkweed and buckwheat also provided nectar to a wide range of insect pollinators. Monarch butterflies fed on milkweed. When placed between peanut and cotton, a strip of soybean was an effective trap crop for cotton, reducing economic damage. Incorporation of buckwheat near soybean enhanced parasitism of Euschistus servus (Say eggs by Telenomus podisi Ashmead in cotton. In conclusion, nectar provision enhances biocontrol of stink bugs, acts together with other management tactics for stink bug control, and aids in conservation of natural enemies, insect pollinators, and the monarch butterfly.

WYSIWIB: A Declarative Approach to Finding Protocols and Bugs in Linux Code

DEFF Research Database (Denmark)

Lawall, Julia Laetitia; Brunel, Julien Pierre Manuel; Hansen, Rene Rydhof

2008-01-01

the tools to be able to find specific kinds of bugs. In this paper, we propose a declarative approach based on a control-flow based program search engine. Our approach is WYSIWIB (What You See Is Where It Bugs), since the programmer is able to express specifications for protocol and bug finding using...

Histamine as an emergent indoor contaminant: Accumulation and persistence in bed bug infested homes.

Directory of Open Access Journals (Sweden)

Zachary C DeVries

Full Text Available Histamine is used in bronchial and dermal provocation, but it is rarely considered an environmental risk factor in allergic disease. Because bed bugs defecate large amounts of histamine as a component of their aggregation pheromone, we sought to determine if histamine accumulates in household dust in bed bug infested homes, and the effects of bed bug eradication with spatial heat on histamine levels in dust. We collected dust in homes and analyzed for histamine before, and up to three months after bed bug eradication. Histamine levels in bed bug infested homes were remarkably high (mean = 54.6±18.9 μg/100 mg of sieved household dust and significantly higher than in control homes not infested with bed bugs (mean < 2.5±1.9 μg/100 mg of sieved household dust. Heat treatments that eradicated the bed bug infestations failed to reduce histamine levels, even three months after treatment. We report a clear association between histamine levels in household dust and bed bug infestations. The high concentrations, persistence, and proximity to humans during sleep suggest that bed bug-produced histamine may represent an emergent contaminant and pose a serious health risk in the indoor environment.

How Does Bug-Handling Effort Differ Among Different Programming Languages?

OpenAIRE

Zhang, Jie; Li, Feng; Hao, Dan; Wang, Meng; Zhang, Lu

2018-01-01

Handling bugs is an essential part of software development. The impact of programming language on this task has long been a topic of much debate. For example, some people hold the view that bugs in Python are easy to handle because its code is easy to read and understand, while some others believe the absence of static typing in Python will lead to higher bug-handling effort. This paper presents the first large-scale study to investigate whether the ecosystems of different (categories of) pro...

Insects: Bugged Out!

Science.gov (United States)

Piehl, Kathy

2011-01-01

Insects really need no introduction. They have lived on earth much longer than humans and vastly outnumber people and all other animal species combined. People encounter them daily in their houses and yards. Yet, when children want to investigate insects, books can help them start their explorations. "Paleo Bugs" carries readers back to the time…

Flexible Connectors between Capsomer Subunits that Regulate Capsid Assembly.

Science.gov (United States)

Hasek, Mary L; Maurer, Joshua B; Hendrix, Roger W; Duda, Robert L

2017-08-04

Viruses build icosahedral capsids of specific size and shape by regulating the spatial arrangement of the hexameric and pentameric protein capsomers in the growing shell during assembly. In the T=7 capsids of Escherichia coli bacteriophage HK97 and other phages, 60 capsomers are hexons, while the rest are pentons that are correctly positioned during assembly. Assembly of the HK97 capsid to the correct size and shape has been shown to depend on specific ionic contacts between capsomers. We now describe additional ionic interactions within capsomers that also regulate assembly. Each is between the long hairpin, the "E-loop," that extends from one subunit to the adjacent subunit within the same capsomer. Glutamate E153 on the E-loop and arginine R210 on the adjacent subunit's backbone alpha-helix form salt bridges in hexamers and pentamers. Mutations that disrupt these salt bridges were lethal for virus production, because the mutant proteins assembled into tubes or sheets instead of capsids. X-ray structures show that the E153-R210 links are flexible and maintained during maturation despite radical changes in capsomer shape. The E153-R210 links appear to form early in assembly to enable capsomers to make programmed changes in their shape during assembly. The links also prevent flattening of capsomers and premature maturation. Mutant phenotypes and modeling support an assembly model in which flexible E153-R210 links mediate capsomer shape changes that control where pentons are placed to create normal-sized capsids. The E-loop may be conserved in other systems in order to play similar roles in regulating assembly. Copyright © 2017 Elsevier Ltd. All rights reserved.

A molecular thermodynamic model for the stability of hepatitis B capsids

Science.gov (United States)

Kim, Jehoon; Wu, Jianzhong

2014-06-01

Self-assembly of capsid proteins and genome encapsidation are two critical steps in the life cycle of most plant and animal viruses. A theoretical description of such processes from a physiochemical perspective may help better understand viral replication and morphogenesis thus provide fresh insights into the experimental studies of antiviral strategies. In this work, we propose a molecular thermodynamic model for predicting the stability of Hepatitis B virus (HBV) capsids either with or without loading nucleic materials. With the key components represented by coarse-grained thermodynamic models, the theoretical predictions are in excellent agreement with experimental data for the formation free energies of empty T4 capsids over a broad range of temperature and ion concentrations. The theoretical model predicts T3/T4 dimorphism also in good agreement with the capsid formation at in vivo and in vitro conditions. In addition, we have studied the stability of the viral particles in response to physiological cellular conditions with the explicit consideration of the hydrophobic association of capsid subunits, electrostatic interactions, molecular excluded volume effects, entropy of mixing, and conformational changes of the biomolecular species. The course-grained model captures the essential features of the HBV nucleocapsid stability revealed by recent experiments.

A molecular thermodynamic model for the stability of hepatitis B capsids

Energy Technology Data Exchange (ETDEWEB)

Kim, Jehoon; Wu, Jianzhong, E-mail: jwu@engr.ucr.edu [Department of Chemical and Environmental Engineering, University of California, Riverside, California 92521 (United States)

2014-06-21

Self-assembly of capsid proteins and genome encapsidation are two critical steps in the life cycle of most plant and animal viruses. A theoretical description of such processes from a physiochemical perspective may help better understand viral replication and morphogenesis thus provide fresh insights into the experimental studies of antiviral strategies. In this work, we propose a molecular thermodynamic model for predicting the stability of Hepatitis B virus (HBV) capsids either with or without loading nucleic materials. With the key components represented by coarse-grained thermodynamic models, the theoretical predictions are in excellent agreement with experimental data for the formation free energies of empty T4 capsids over a broad range of temperature and ion concentrations. The theoretical model predicts T3/T4 dimorphism also in good agreement with the capsid formation at in vivo and in vitro conditions. In addition, we have studied the stability of the viral particles in response to physiological cellular conditions with the explicit consideration of the hydrophobic association of capsid subunits, electrostatic interactions, molecular excluded volume effects, entropy of mixing, and conformational changes of the biomolecular species. The course-grained model captures the essential features of the HBV nucleocapsid stability revealed by recent experiments.

Mutation of a Conserved Nuclear Export Sequence in Chikungunya Virus Capsid Protein Disrupts Host Cell Nuclear Import.

Science.gov (United States)

Jacobs, Susan C; Taylor, Adam; Herrero, Lara J; Mahalingam, Suresh; Fazakerley, John K

2017-10-20

Transmitted by mosquitoes; chikungunya virus (CHIKV) is responsible for frequent outbreaks of arthritic disease in humans. CHIKV is an arthritogenic alphavirus of the Togaviridae family. Capsid protein, a structural protein encoded by the CHIKV RNA genome, is able to translocate to the host cell nucleus. In encephalitic alphaviruses nuclear translocation induces host cell shut off; however, the role of capsid protein nuclear localisation in arthritogenic alphaviruses remains unclear. Using replicon systems, we investigated a nuclear export sequence (NES) in the N-terminal region of capsid protein; analogous to that found in encephalitic alphavirus capsid but uncharacterised in CHIKV. The chromosomal maintenance 1 (CRM1) export adaptor protein mediated CHIKV capsid protein export from the nucleus and a region within the N-terminal part of CHIKV capsid protein was required for active nuclear targeting. In contrast to encephalitic alphaviruses, CHIKV capsid protein did not inhibit host nuclear import; however, mutating the NES of capsid protein (∆NES) blocked host protein access to the nucleus. Interactions between capsid protein and the nucleus warrant further investigation.

A methodology for quantitatively managing the bug fixing process using Mahalanobis Taguchi system

Directory of Open Access Journals (Sweden)

Boby John

2015-12-01

Full Text Available The controlling of bug fixing process during the system testing phase of software development life cycle is very important for fixing all the detected bugs within the scheduled time. The presence of open bugs often delays the release of the software or result in releasing the software with compromised functionalities. These can lead to customer dissatisfaction, cost overrun and eventually the loss of market share. In this paper, the authors propose a methodology to quantitatively manage the bug fixing process during system testing. The proposed methodology identifies the critical milestones in the system testing phase which differentiates the successful projects from the unsuccessful ones using Mahalanobis Taguchi system. Then a model is developed to predict whether a project is successful or not with the bug fix progress at critical milestones as control factors. Finally the model is used to control the bug fixing process. It is found that the performance of the proposed methodology using Mahalanobis Taguchi system is superior to the models developed using other multi-dimensional pattern recognition techniques. The proposed methodology also reduces the number of control points providing the managers with more options and flexibility to utilize the bug fixing resources across system testing phase. Moreover the methodology allows the mangers to carry out mid- course corrections to bring the bug fixing process back on track so that all the detected bugs can be fixed on time. The methodology is validated with eight new projects and the results are very encouraging.

Pheromones of milkweed bugs (Heteroptera: Lygaeidae) attract wayward plant bugs: Phytocoris mirid sex pheromone.

Science.gov (United States)

Zhang, Qing-He; Aldrich, Jeffrey R

2003-08-01

The synthetic aggregation pheromone of the large milkweed bug, Oncopeltus fasciatus (Dallas) (Lygaeinae), also attracted males of the plant bug, Phytocoris difficilis Knight (Miridae). Field testing partial blends against the six-component blend comprising the Oncopeltus pheromone showed that cross-attraction of P. difficilis males was due to synergism between (E)-2-octenyl acetate and (E,E)-2,4-hexadienyl acetate. Hexyl acetate was abundant in the metathoracic scent gland (MSG) secretion of P. difficilis males, but because female P. difficilis could not initially be found in the field, further combinatorial tests were guided by prior research on the pheromones of two Phytocoris species in the western United States. The combination of hexyl, (E)-2-hexenyl, and (E)-2-octenyl acetates was as attractive to P. difficilis males as the milkweed bug pheromone, yet no milkweed bugs were drawn to this blend. Gas chromatographic (GC)-electroantennographic detection (EAD) and GC-mass spectrometric (MS) analyses of female P. difficilis MSGs determined that their secretion contained predominantly hexyl, (E)-2-hexenyl, and (E)-2-octenyl acetates (all strongly EAD-active)-the latter two compounds found only in trace amounts from males-plus five minor female-specific compounds, three of which were EAD-active. (E,E)-2,4-Hexadienyl acetate was not detected from P. difficilis females or males. The blend of the three major components, hexyl, (E)-2-hexenyl, and (E)-2-octenyl acetates (2:1.5:1 by volume), was as attractive as the blend of all six EAD-active compounds identified from females, indicating that this ternary blend constitutes the sex pheromone of P. difficilis. Hexyl acetate with (E)-2-octenyl acetate also attracted males of another species, P. breviusculus Reuter, but addition of (E)-2-hexenyl acetate and/or (E,E)-2,4-hexadienyl acetate inhibited attraction of P. breviusculus males. Attraction of P. difficilis males occurred mainly during the first half of scotophase. The

How viral capsids adapt to mismatched cargoes—identifying mechanisms of morphology control with simulations

Science.gov (United States)

Elrad, Oren

2009-03-01

During the replication of many viruses, hundreds to thousands of protein subunits assemble around the viral nucleic acid to form a protein shell called a capsid. Most viruses form one particular structure with astonishing fidelity; yet, recent experiments demonstrate that capsids can assemble with different sizes and morphologies to accommodate nucleic acids or other cargoes such as functionalized nanoparticles. In this talk, we will explore the mechanisms of simultaneous assembly and cargo encapsidation with a computational model that describes the assembly of icosahedral capsids around functionalized nanoparticles. With this model, we find parameter values for which subunits faithfully form empty capsids with a single morphology, but adaptively assemble into different icosahedral morphologies around nanoparticles with different diameters. Analyzing trajectories in which adaptation is or is not successful sheds light on the mechanisms by which capsid morphology may be controlled in vitro and in vivo, and suggests experiments to test these mechanisms. We compare the simulation results to recent experiments in which Brome Mosaic Virus capsid proteins assemble around functionalized nanoparticles, and describe how future experiments can test the model predictions.

Histamine as an emergent indoor contaminant: Accumulation and persistence in bed bug infested homes.

Science.gov (United States)

DeVries, Zachary C; Santangelo, Richard G; Barbarin, Alexis M; Schal, Coby

2018-01-01

Histamine is used in bronchial and dermal provocation, but it is rarely considered an environmental risk factor in allergic disease. Because bed bugs defecate large amounts of histamine as a component of their aggregation pheromone, we sought to determine if histamine accumulates in household dust in bed bug infested homes, and the effects of bed bug eradication with spatial heat on histamine levels in dust. We collected dust in homes and analyzed for histamine before, and up to three months after bed bug eradication. Histamine levels in bed bug infested homes were remarkably high (mean = 54.6±18.9 μg/100 mg of sieved household dust) and significantly higher than in control homes not infested with bed bugs (mean emergent contaminant and pose a serious health risk in the indoor environment.

Cleavage sites within the poliovirus capsid protein precursors

International Nuclear Information System (INIS)

Larsen, G.R.; Anderson, C.W.; Dorner, A.J.; Semler, B.L.; Wimmer, E.

1982-01-01

Partial amino-terminal sequence analysis was performed on radiolabeled poliovirus capsid proteins VP1, VP2, and VP3. A computer-assisted comparison of the amino acid sequences obtained with that predicted by the nucleotide sequence of the poliovirus genome allows assignment of the amino terminus of each capsid protein to a unique position within the virus polyprotein. Sequence analysis of trypsin-digested VP4, which has a blocked amino terminus, demonstrates that VP4 is encoded at or very near to the amino terminus of the polyprotein. The gene order of the capsid proteins is VP4-VP2-VP3-VP1. Cleavage of VP0 to VP4 and VP2 is shown to occur between asparagine and serine, whereas the cleavages that separate VP2/VP3 and VP3/VP1 occur between glutamine and glycine residues. This finding supports the hypothesis that the cleavage of VP0, which occurs during virion morphogenesis, is distinct from the cleavages that separate functional regions of the polyprotein

Overcoming preexisting humoral immunity to AAV using capsid decoys.

Science.gov (United States)

Mingozzi, Federico; Anguela, Xavier M; Pavani, Giulia; Chen, Yifeng; Davidson, Robert J; Hui, Daniel J; Yazicioglu, Mustafa; Elkouby, Liron; Hinderer, Christian J; Faella, Armida; Howard, Carolann; Tai, Alex; Podsakoff, Gregory M; Zhou, Shangzhen; Basner-Tschakarjan, Etiena; Wright, John Fraser; High, Katherine A

2013-07-17

Adeno-associated virus (AAV) vectors delivered through the systemic circulation successfully transduce various target tissues in animal models. However, similar attempts in humans have been hampered by the high prevalence of neutralizing antibodies to AAV, which completely block vector transduction. We show in both mouse and nonhuman primate models that addition of empty capsid to the final vector formulation can, in a dose-dependent manner, adsorb these antibodies, even at high titers, thus overcoming their inhibitory effect. To further enhance the safety of the approach, we mutated the receptor binding site of AAV2 to generate an empty capsid mutant that can adsorb antibodies but cannot enter a target cell. Our work suggests that optimizing the ratio of full/empty capsids in the final formulation of vector, based on a patient's anti-AAV titers, will maximize the efficacy of gene transfer after systemic vector delivery.

General Model for Retroviral Capsid Pattern Recognition by TRIM5 Proteins.

Science.gov (United States)

Wagner, Jonathan M; Christensen, Devin E; Bhattacharya, Akash; Dawidziak, Daria M; Roganowicz, Marcin D; Wan, Yueping; Pumroy, Ruth A; Demeler, Borries; Ivanov, Dmitri N; Ganser-Pornillos, Barbie K; Sundquist, Wesley I; Pornillos, Owen

2018-02-15

Restriction factors are intrinsic cellular defense proteins that have evolved to block microbial infections. Retroviruses such as HIV-1 are restricted by TRIM5 proteins, which recognize the viral capsid shell that surrounds, organizes, and protects the viral genome. TRIM5α uses a SPRY domain to bind capsids with low intrinsic affinity ( K D of >1 mM) and therefore requires higher-order assembly into a hexagonal lattice to generate sufficient avidity for productive capsid recognition. TRIMCyp, on the other hand, binds HIV-1 capsids through a cyclophilin A domain, which has a well-defined binding site and higher affinity ( K D of ∼10 μM) for isolated capsid subunits. Therefore, it has been argued that TRIMCyp proteins have dispensed with the need for higher-order assembly to function as antiviral factors. Here, we show that, consistent with its high degree of sequence similarity with TRIM5α, the TRIMCyp B-box 2 domain shares the same ability to self-associate and facilitate assembly of a TRIMCyp hexagonal lattice that can wrap about the HIV-1 capsid. We also show that under stringent experimental conditions, TRIMCyp-mediated restriction of HIV-1 is indeed dependent on higher-order assembly. Both forms of TRIM5 therefore use the same mechanism of avidity-driven capsid pattern recognition. IMPORTANCE Rhesus macaques and owl monkeys are highly resistant to HIV-1 infection due to the activity of TRIM5 restriction factors. The rhesus macaque TRIM5α protein blocks HIV-1 through a mechanism that requires self-assembly of a hexagonal TRIM5α lattice around the invading viral core. Lattice assembly amplifies very weak interactions between the TRIM5α SPRY domain and the HIV-1 capsid. Assembly also promotes dimerization of the TRIM5α RING E3 ligase domain, resulting in synthesis of polyubiquitin chains that mediate downstream steps of restriction. In contrast to rhesus TRIM5α, the owl monkey TRIM5 homolog, TRIMCyp, binds isolated HIV-1 CA subunits much more tightly

Antigenic structure of the capsid protein of rabbit haemorrhagic disease virus

DEFF Research Database (Denmark)

Martinez-Torrecuadrada, Jorge L.; Cortes, Elena; Vela, Carmen

1998-01-01

Rabbit haemorrhagic disease virus (RHDV) causes an important disease in rabbits. The virus capsid is composed of a single 60 kDa protein. The capsid protein gene was cloned in Escherichia coli using the pET3 system, and the antigenic structure of RHDV VP60 was dissected using 11 monoclonal...

Structural Characterization of H-1 Parvovirus: Comparison of Infectious Virions to Empty Capsids

Science.gov (United States)

Halder, Sujata; Nam, Hyun-Joo; Govindasamy, Lakshmanan; Vogel, Michèle; Dinsart, Christiane; Salomé, Nathalie; McKenna, Robert

2013-01-01

The structure of single-stranded DNA (ssDNA) packaging H-1 parvovirus (H-1PV), which is being developed as an antitumor gene delivery vector, has been determined for wild-type (wt) virions and noninfectious (empty) capsids to 2.7- and 3.2-Å resolution, respectively, using X-ray crystallography. The capsid viral protein (VP) structure consists of an α-helix and an eight-stranded anti-parallel β-barrel with large loop regions between the strands. The β-barrel and loops form the capsid core and surface, respectively. In the wt structure, 600 nucleotides are ordered in an interior DNA binding pocket of the capsid. This accounts for ∼12% of the H-1PV genome. The wt structure is identical to the empty capsid structure, except for side chain conformation variations at the nucleotide binding pocket. Comparison of the H-1PV nucleotides to those observed in canine parvovirus and minute virus of mice, two members of the genus Parvovirus, showed both similarity in structure and analogous interactions. This observation suggests a functional role, such as in capsid stability and/or ssDNA genome recognition for encapsulation. The VP structure differs from those of other parvoviruses in surface loop regions that control receptor binding, tissue tropism, pathogenicity, and antibody recognition, including VP sequences reported to determine tumor cell tropism for oncotropic rodent parvoviruses. These structures of H-1PV provide insight into structural features that dictate capsid stabilization following genome packaging and three-dimensional information applicable for rational design of tumor-targeted recombinant gene delivery vectors. PMID:23449783

Aggregation and Association of NDVI, Boll Injury, and Stink Bugs in North Carolina Cotton.

Science.gov (United States)

Reisig, Dominic D; Reay-Jones, F P F; Meijer, A D

2015-01-01

Sampling of herbivorous stink bugs in southeastern U.S. cotton remains problematic. Remote sensing was explored to improve sampling of these pests and associated boll injury. Two adjacent 14.5-ha cotton fields were grid sampled in 2011 and 2012 by collecting stink bug adults and bolls every week during the third, fourth, and fifth weeks of bloom. Satellite remote sensing data were collected during the third week of bloom during both years, and normalized difference vegetation index (NDVI) values were calculated. Stink bugs were spatially aggregated on the third week of bloom in 2011. Boll injury from stink bugs was spatially aggregated during the fourth week of bloom in 2012. The NDVI values were aggregated during both years. There was a positive association and correlation between stink bug numbers and NDVI values, as well as injured bolls and NDVI values, during the third week of bloom in 2011. During the third week of bloom in 2012, NDVI values were negatively correlated with stink bug numbers. During the fourth week of bloom in 2011, stink bug numbers and boll injury were both positively associated and correlated with NDVI values. During the fourth week of bloom in 2012, stink bugs were negatively correlated with NDVI values, and boll injury was negatively associated and correlated with NDVI values. This study suggests the potential of remote sensing as a tool to assist with sampling stink bugs in cotton, although more research is needed using NDVI and other plant measurements to predict stink bug injury. © The Author 2015. Published by Oxford University Press on behalf of the Entomological Society of America.

WYSIWYB: A Declarative Approach to Finding API Protocols and Bugs in Linux Code

DEFF Research Database (Denmark)

Lawall, Julia; Lawall, Julia; Palix, Nicolas

2009-01-01

the tools to be able to find specific kinds of bugs. In this paper, we propose a declarative approach based on a control-flow based program search engine. Our approach is WYSIWIB (What You See Is Where It Bugs), since the programmer is able to express specifications for protocol and bug finding using...

WYSIWIB: A Declarative Approach to Finding API Protocols and Bugs in Linux Code

DEFF Research Database (Denmark)

Lawall, Julia; Brunel, Julien Pierre Manuel; Palix, Nicolas Jean-Michel

2009-01-01

the tools on specific kinds of bugs and to relate the results to patterns in the source code. We propose a declarative approach to bug finding in Linux OS code using a control-flow based program search engine. Our approach is WYSIWIB (What You See Is Where It Bugs), since the programmer expresses...

Traveler's Health: Avoid Bug Bites

Science.gov (United States)

... Safety Road Safety - 8 Steps MERS Health Advisory poster MERS Pictogram CDC Guide for Healthy Travel Website ... other accommodations that are air conditioned or have good window and door screens so bugs can’t ...

Getting Rid of Bed Bugs

Science.gov (United States)

... Directory Planning, Budget and Results Jobs and Internships Headquarters Offices Regional Offices Labs and Research Centers Bed ... to be careful in how you select a company. Related Information Collaborative Strategy on Bed Bugs - highlights ...

EPA-Registered Bed Bug Products

Science.gov (United States)

... rest or hide in hampers, bed frames, even furniture); Failing to treat adjacent areas where bed bugs ... to work (some pesticides, such as desiccators or growth regulators, may be very effective but take some ...

Cases of bed bug (Cimex lectularius infestations in Northwest Italy

Directory of Open Access Journals (Sweden)

Federica Giorda

2013-12-01

Full Text Available Bed bugs (Cimex lectularius have been a common problem for humans for at least 3,500 years and in Europe their presence was endemic until the end of World War II, when infestations began to decrease. However, since the beginning of the 21st century new cases of infestations have been reported in developed countries. Many theories have been put forward to explain this change of direction, but none has been scientifically proven. The aim of this study is to provide some reports of bed bug infestations in Northern Italy (Liguria, Piedmont and Aosta valley regions and a brief summary about their identification, clinical significance, bioecology and control. From 2008 to date, 17 bed bug infestations were identified in Northwest Italy. Knowledge about the presence and distribution of bed bugs in Italy is scanty, prior to this work only 2 studies reported the comeback of these arthropods in the Italian territory; further investigations would be necessary to better understand the current situation.

Functional characterization of Kaposi's sarcoma-associated herpesvirus small capsid protein by bacterial artificial chromosome-based mutagenesis

International Nuclear Information System (INIS)

Sathish, Narayanan; Yuan Yan

2010-01-01

A systematic investigation of interactions amongst KSHV capsid proteins was undertaken in this study to comprehend lesser known KSHV capsid assembly mechanisms. Interestingly the interaction patterns of the KSHV small capsid protein, ORF65 suggested its plausible role in viral capsid assembly pathways. Towards further understanding this, ORF65-null recombinant mutants (BAC-Δ65 and BAC-stop65) employing a bacterial artificial chromosome (BAC) system were generated. No significant difference was found in both overall viral gene expression and lytic DNA replication between stable monolayers of 293T-BAC36 (wild-type) and 293T-BAC-ORF65-null upon induction with 12-O-tetradecanoylphorbol-13-acetate, though the latter released 30-fold fewer virions to the medium than 293T-BAC36 cells. Sedimentation profiles of capsid proteins of ORF65-null recombinant mutants were non-reflective of their organization into the KSHV capsids and were also undetectable in cytoplasmic extracts compared to noticeable levels in nuclear extracts. These observations collectively suggested the pivotal role of ORF65 in the KSHV capsid assembly processes.

Pt, Co–Pt and Fe–Pt alloy nanoclusters encapsulated in virus capsids

International Nuclear Information System (INIS)

Okuda, M; Eloi, J-C; Jones, S E Ward; Schwarzacher, W; Verwegen, M; Cornelissen, J J L M

2016-01-01

Nanostructured Pt-based alloys show great promise, not only for catalysis but also in medical and magnetic applications. To extend the properties of this class of materials, we have developed a means of synthesizing Pt and Pt-based alloy nanoclusters in the capsid of a virus. Pure Pt and Pt-alloy nanoclusters are formed through the chemical reduction of [PtCl 4 ] − by NaBH 4 with/without additional metal ions (Co or Fe). The opening and closing of the ion channels in the virus capsid were controlled by changing the pH and ionic strength of the solution. The size of the nanoclusters is limited to 18 nm by the internal diameter of the capsid. Their magnetic properties suggest potential applications in hyperthermia for the Co–Pt and Fe–Pt magnetic alloy nanoclusters. This study introduces a new way to fabricate size-restricted nanoclusters using virus capsid. (paper)

Intracellular Distribution of Capsid-Associated pUL77 of Human Cytomegalovirus and Interactions with Packaging Proteins and pUL93.

Science.gov (United States)

Köppen-Rung, Pánja; Dittmer, Alexandra; Bogner, Elke

2016-07-01

DNA packaging into procapsids is a common multistep process during viral maturation in herpesviruses. In human cytomegalovirus (HCMV), the proteins involved in this process are terminase subunits pUL56 and pUL89, which are responsible for site-specific cleavage and insertion of the DNA into the procapsid via portal protein pUL104. However, additional viral proteins are required for the DNA packaging process. We have shown previously that the plasmid that encodes capsid-associated pUL77 encodes another potential player during capsid maturation. Pulse-chase experiments revealed that pUL77 is stably expressed during HCMV infection. Time course analysis demonstrated that pUL77 is expressed in the early late part of the infectious cycle. The sequence of pUL77 was analyzed to find nuclear localization sequences (NLSs), revealing monopartite NLSm at the N terminus and bipartite NLSb in the middle of pUL77. The potential NLSs were inserted into plasmid pHM829, which encodes a chimeric protein with β-galactosidase and green fluorescent protein. In contrast to pUL56, neither NLSm nor NLSb was sufficient for nuclear import. Furthermore, we investigated by coimmunoprecipitation whether packaging proteins, as well as pUL93, the homologue protein of herpes simplex virus 1 pUL17, are interaction partners of pUL77. The interactions between pUL77 and packaging proteins, as well as pUL93, were verified. We showed that the capsid-associated pUL77 is another potential player during capsid maturation of HCMV. Protein UL77 (pUL77) is a conserved core protein of HCMV. This study demonstrates for the first time that pUL77 has early-late expression kinetics during the infectious cycle and an intrinsic potential for nuclear translocation. According to its proposed functions in stabilization of the capsid and anchoring of the encapsidated DNA during packaging, interaction with further DNA packaging proteins is required. We identified physical interactions with terminase subunits pUL56 and p

The Assembly-Activating Protein Promotes Stability and Interactions between AAV’s Viral Proteins to Nucleate Capsid Assembly

Directory of Open Access Journals (Sweden)

Anna C. Maurer

2018-05-01

Full Text Available Summary: The adeno-associated virus (AAV vector is a preferred delivery platform for in vivo gene therapy. Natural and engineered variations of the AAV capsid affect a plurality of phenotypes relevant to gene therapy, including vector production and host tropism. Fundamental to these aspects is the mechanism of AAV capsid assembly. Here, the role of the viral co-factor assembly-activating protein (AAP was evaluated in 12 naturally occurring AAVs and 9 putative ancestral capsid intermediates. The results demonstrate increased capsid protein stability and VP-VP interactions in the presence of AAP. The capsid’s dependence on AAP can be partly overcome by strengthening interactions between monomers within the assembly, as illustrated by the transfer of a minimal motif defined by a phenotype-to-phylogeny mapping method. These findings suggest that the emergence of AAP within the Dependovirus genus relaxes structural constraints on AAV assembly in favor of increasing the degrees of freedom for the capsid to evolve. : Maurer et al. describe a phenotype-to-phylogeny mapping strategy correlating phenotypic variation in AAVs to a reconstructed phylogeny, revealing capsid structure-function relationships relevant to that phenotype. Dependence on the viral co-factor AAP for capsid assembly is examined, and capsid functional motifs, in addition to mechanistic roles of AAP, are elucidated. Keywords: AAV, AAP, adeno-associated virus, capsid assembly, manufacturing, capsid, vector engineering, structure-function, gene therapy

An automated approach for finding variable-constant pairing bugs

DEFF Research Database (Denmark)

Lawall, Julia; Lo, David

2010-01-01

program-analysis and data-mining based approach to identify the uses of named constants and to identify anomalies in these uses.  We have applied our approach to a recent version of the Linux kernel and have found a number of bugs affecting both correctness and software maintenance.  Many of these bugs...... have been validated by the Linux developers....

Improvement on the accuracy of beam bugs in linear induction accelerator

International Nuclear Information System (INIS)

Xie Yutong; Dai Zhiyong; Han Qing

2002-01-01

In linear induction accelerator the resistive wall monitors known as 'beam bugs' have been used as essential diagnostics of beam current and location. The author presents a new method that can improve the accuracy of these beam bugs used for beam position measurements. With a fine beam simulation set, this method locates the beam position with an accuracy of 0.02 mm and thus can scale the beam bugs very well. Experiment results prove that the precision of beam position measurements can reach submillimeter degree

Protruding Features of Viral Capsids Are Clustered on Icosahedral Great Circles.

Directory of Open Access Journals (Sweden)

David P Wilson

Full Text Available Spherical viruses are remarkably well characterized by the Triangulation (T number developed by Casper and Klug. The T-number specifies how many viral capsid proteins are required to cover the virus, as well as how they are further subdivided into pentamer and hexamer subunits. The T-number however does not constrain the orientations of these proteins within the subunits or dictate where the proteins should place their protruding features. These protrusions often take the form of loops, spires and helices, and are significant because they aid in stability of the capsid as well as recognition by the host organism. Until now there has be no overall understanding of the placement of protrusions for spherical viruses, other than they have icosahedral symmetry. We constructed a set of gauge points based upon the work affine extensions of Keef and Twarock, which have fixed relative angular locations with which to measure the locations of these features. This work adds a new element to our understanding of the geometric arrangement of spherical viral capsid proteins; chiefly that the locations of protruding features are not found stochastically distributed in an icosahedral manner across the viral surface, but instead these features are found only in specific locations along the 15 icosahedral great circles. We have found that this result holds true as the T number and viral capsids size increases, suggesting an underlying geometric constraint on their locations. This is in spite of the fact that the constraints on the pentamers and hexamer orientations change as a function of T-number, as you need to accommodate more hexamers in the same solid angle between pentamers. The existence of this angular constraint of viral capsids suggests that there is a fitness or energetic benefit to the virus placing its protrusions in this manner. This discovery may have profound impacts on identifying and eliminating viral pathogens, understanding evolutionary

Relevance of capsid structure in the buckling and maturation of spherical viruses

International Nuclear Information System (INIS)

Aznar, María; Luque, Antoni; Reguera, David

2012-01-01

The shape and mechanical properties of viral capsids play an important role in several biological processes during the virus life cycle. In particular, to become infective, many viruses require a maturation stage where the capsid undergoes a buckling transition, from an initial spherical procapsid into a final icosahedral faceted shell. Here we study, using a minimal physical model, how the capsid shape and the buckling transition depend on the triangulation number T and the icosahedral class P of the virus structure. We find that, for small shells, capsids with P = 1 are most likely to produce polyhedral shapes that minimize their energy and accumulated stress, whereas viruses with P = 3 prefer to remain spherical. For big capsids, all shells are more stable adopting an icosahedral shape, in agreement with continuum elastic theory. Moreover, spherical viruses show a buckling transition to polyhedral shells under expansion, in consonance with virus maturation. The resulting icosahedral shell is mechanically stiffer, tolerates larger expansions and withstands higher internal pressures before failing, which could explain why some dsDNA viruses, which rely on the pressurization of their genetic material to facilitate the infection, undergo a buckling transition. We emphasize that the results are general and could also be applied to non-biological systems. (paper)

Assembly of recombinant Israeli Acute Paralysis Virus capsids.

Directory of Open Access Journals (Sweden)

Junyuan Ren

Full Text Available The dicistrovirus Israeli Acute Paralysis Virus (IAPV has been implicated in the worldwide decline of honey bees. Studies of IAPV and many other bee viruses in pure culture are restricted by available isolates and permissive cell culture. Here we show that coupling the IAPV major structural precursor protein ORF2 to its cognate 3C-like processing enzyme results in processing of the precursor to the individual structural proteins in a number of insect cell lines following expression by a recombinant baculovirus. The efficiency of expression is influenced by the level of IAPV 3C protein and moderation of its activity is required for optimal expression. The mature IAPV structural proteins assembled into empty capsids that migrated as particles on sucrose velocity gradients and showed typical dicistrovirus like morphology when examined by electron microscopy. Monoclonal antibodies raised to recombinant capsids were configured into a diagnostic test specific for the presence of IAPV. Recombinant capsids for each of the many bee viruses within the picornavirus family may provide virus specific reagents for the on-going investigation of the causes of honeybee loss.

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.

Inner tegument proteins of Herpes Simplex Virus are sufficient for intracellular capsid motility in neurons but not for axonal targeting

Science.gov (United States)

Müller, Oliver; Ivanova, Lyudmila; Bialy, Dagmara; Pohlmann, Anja; Binz, Anne; Hegemann, Maike; Viejo-Borbolla, Abel; Rosenhahn, Bodo; Bauerfeind, Rudolf; Sodeik, Beate

2017-01-01

Upon reactivation from latency and during lytic infections in neurons, alphaherpesviruses assemble cytosolic capsids, capsids associated with enveloping membranes, and transport vesicles harboring fully enveloped capsids. It is debated whether capsid envelopment of herpes simplex virus (HSV) is completed in the soma prior to axonal targeting or later, and whether the mechanisms are the same in neurons derived from embryos or from adult hosts. We used HSV mutants impaired in capsid envelopment to test whether the inner tegument proteins pUL36 or pUL37 necessary for microtubule-mediated capsid transport were sufficient for axonal capsid targeting in neurons derived from the dorsal root ganglia of adult mice. Such neurons were infected with HSV1-ΔUL20 whose capsids recruited pUL36 and pUL37, with HSV1-ΔUL37 whose capsids associate only with pUL36, or with HSV1-ΔUL36 that assembles capsids lacking both proteins. While capsids of HSV1-ΔUL20 were actively transported along microtubules in epithelial cells and in the somata of neurons, those of HSV1-ΔUL36 and -ΔUL37 could only diffuse in the cytoplasm. Employing a novel image analysis algorithm to quantify capsid targeting to axons, we show that only a few capsids of HSV1-ΔUL20 entered axons, while vesicles transporting gD utilized axonal transport efficiently and independently of pUL36, pUL37, or pUL20. Our data indicate that capsid motility in the somata of neurons mediated by pUL36 and pUL37 does not suffice for targeting capsids to axons, and suggest that capsid envelopment needs to be completed in the soma prior to targeting of herpes simplex virus to the axons, and to spreading from neurons to neighboring cells. PMID:29284065

Oral Administration of Astrovirus Capsid Protein Is Sufficient To Induce Acute Diarrhea In Vivo

Directory of Open Access Journals (Sweden)

Victoria A. Meliopoulos

2016-11-01

Full Text Available The disease mechanisms associated with the onset of astrovirus diarrhea are unknown. Unlike other enteric virus infections, astrovirus infection is not associated with an inflammatory response or cellular damage. In vitro studies in differentiated Caco-2 cells demonstrated that human astrovirus serotype 1 (HAstV-1 capsid protein alone disrupts the actin cytoskeleton and tight junction complex, leading to increased epithelial barrier permeability. In this study, we show that oral administration of purified recombinant turkey astrovirus 2 (TAstV-2 capsid protein results in acute diarrhea in a dose- and time-dependent manner in turkey poults. Similarly to that induced by infectious virus, TAstV-2 capsid-induced diarrhea was independent of inflammation or histological changes but was associated with increased intestinal barrier permeability, as well as redistribution of sodium hydrogen exchanger 3 (NHE3 from the membrane to the cytoplasm of the intestinal epithelium. Unlike other viral enterotoxins that have been identified, astrovirus capsid induces diarrhea after oral administration, reproducing the natural route of infection and demonstrating that ingestion of intact noninfectious capsid protein may be sufficient to provoke acute diarrhea. Based on these data, we hypothesize that the astrovirus capsid acts like an enterotoxin and induces intestinal epithelial barrier dysfunction.

Hybridization and adaptation to introduced balloon vines in an Australian soapberry bug.

Science.gov (United States)

Andres, J A; Thampy, P R; Mathieson, M T; Loye, J; Zalucki, M P; Dingle, H; Carroll, S P

2013-12-01

Contemporary adaptation of plant feeding insects to introduced hosts provides clear cases of ecologically based population divergence. In most cases the mechanisms permitting rapid differentiation are not well known. Here we study morphological and genetic variation associated with recent shifts by the Australian soapberry bug Leptocoris tagalicus onto two naturalized Neotropical balloon vines, Cardiospermum halicacabum and C. grandiflorum that differ in time since introduction. Our results show that these vines have much larger fruits than the native hosts (Whitewood tree -Atalaya hemiglauca- and Woolly Rambutan -Alectryon tomentosus-) and that bugs living on them have evolved significantly longer beaks and new allometries. Genetic analyses of mitochondrial haplotypes and amplified fragment length polymorphic (AFLP) markers indicate that the lineage of bugs on the annual vine C. halicacabum, the older introduction, is intermediate between the two subspecies of L. tagalicus found on native hosts. Moreover, where the annual vine and Whitewood tree co-occur, the morphology and genomic composition of the bugs are similar to those occurring in allopatry. These results show that hybridization provided the genetic elements underlying the strongly differentiated 'Halicacabum bugs'. In contrast, the bugs feeding on the recently introduced perennial balloon vine (C. grandiflorum) showed no evidence of admixture, and are genetically indistinguishable from the nearby populations on a native host. © 2013 John Wiley & Sons Ltd.

Bug tracking and project management system application in an electronic design company

Directory of Open Access Journals (Sweden)

Sadık ARSLAN

2016-05-01

Full Text Available In this study, commercially available Bug Tracking and Management Information Systems has been investigated in a comprehensive manner. The systems that commonly used described in detail. Bug Tracking and Project Management Systems requirements analysis of medium-sized companies and Kentkart Ege Electronic which is an Information Technology company has been made. Obtained by the analysis requirements, the appropriate tools are selected for system application. JIRA that a product of Atlassian company was determined as a Bug Tracking and Project Management application tool. In this study, JIRA system adapted to the requirements, Bug Tracking and Project Management systems is designed in a structure which can be easily used by R&D employees. Cost-Benefit analysis is done and using this project was determined to be quite useful.

How to Find Bed Bugs

Science.gov (United States)

Find and correctly identify an infestation early before it becomes widespread. Look for rusty or reddish stains and pinpoint dark spots on bed sheets or mattresses, and search for bugs near the piping, seams and tags of the mattress and box spring.

Functional requirements of the yellow fever virus capsid protein.

Science.gov (United States)

Patkar, Chinmay G; Jones, Christopher T; Chang, Yu-hsuan; Warrier, Ranjit; Kuhn, Richard J

2007-06-01

Although it is known that the flavivirus capsid protein is essential for genome packaging and formation of infectious particles, the minimal requirements of the dimeric capsid protein for virus assembly/disassembly have not been characterized. By use of a trans-packaging system that involved packaging a yellow fever virus (YFV) replicon into pseudo-infectious particles by supplying the YFV structural proteins using a Sindbis virus helper construct, the functional elements within the YFV capsid protein (YFC) were characterized. Various N- and C-terminal truncations, internal deletions, and point mutations of YFC were analyzed for their ability to package the YFV replicon. Consistent with previous reports on the tick-borne encephalitis virus capsid protein, YFC demonstrates remarkable functional flexibility. Nearly 40 residues of YFC could be removed from the N terminus while the ability to package replicon RNA was retained. Additionally, YFC containing a deletion of approximately 27 residues of the C terminus, including a complete deletion of C-terminal helix 4, was functional. Internal deletions encompassing the internal hydrophobic sequence in YFC were, in general, tolerated to a lesser extent. Site-directed mutagenesis of helix 4 residues predicted to be involved in intermonomeric interactions were also analyzed, and although single mutations did not affect packaging, a YFC with the double mutation of leucine 81 and valine 88 was nonfunctional. The effects of mutations in YFC on the viability of YFV infection were also analyzed, and these results were similar to those obtained using the replicon packaging system, thus underscoring the flexibility of YFC with respect to the requirements for its functioning.

Hepatitis B virus core protein allosteric modulators can distort and disrupt intact capsids.

Science.gov (United States)

Schlicksup, Christopher John; Wang, Joseph Che-Yen; Francis, Samson; Venkatakrishnan, Balasubramanian; Turner, William W; VanNieuwenhze, Michael; Zlotnick, Adam

2018-01-29

Defining mechanisms of direct-acting antivirals facilitates drug development and our understanding of virus function. Heteroaryldihydropyrimidines (HAPs) inappropriately activate assembly of hepatitis B virus (HBV) core protein (Cp), suppressing formation of virions. We examined a fluorophore-labeled HAP, HAP-TAMRA. HAP-TAMRA induced Cp assembly and also bound pre-assembled capsids. Kinetic and spectroscopic studies imply that HAP-binding sites are usually not available but are bound cooperatively. Using cryo-EM, we observed that HAP-TAMRA asymmetrically deformed capsids, creating a heterogeneous array of sharp angles, flat regions, and outright breaks. To achieve high resolution reconstruction (HAP-TAMRA caused quasi-sixfold vertices to become flatter and fivefold more angular. This transition led to asymmetric faceting. That a disordered crosslink could rescue symmetry implies that capsids have tensegrity properties. Capsid distortion and disruption is a new mechanism by which molecules like the HAPs can block HBV infection. © 2017, Schlicksup et al.

Diversity of some endophytic fungi associated with rice black bug Paraeucosmetus pallicornis on rice plant

OpenAIRE

Nur, Amin; La Daha; Nurariaty, Agus; Ade, Rosmana; Muh., Fadlan

2015-01-01

A new rice insect pest was sighted in some rice producing areas of South Sulawesi Province, Indonesia. This pest is rice black bugs Paraeucosmetus pallicornis. The research aimed to isolation of fungi associated with rice black bugs Paraeucosmetus pallicornis, so as to know the cause of a bitter taste to the rice. The isolation of the fungi consist of three kinds of treatment, namely rice black bugs without sterilization, with sterilization and rice black bugs cut and sterilized. The resul...

Stability of Spatial Distributions of Stink Bugs, Boll Injury, and NDVI in Cotton.

Science.gov (United States)

Reay-Jones, Francis P F; Greene, Jeremy K; Bauer, Philip J

2016-10-01

A 3-yr study was conducted to determine the degree of aggregation of stink bugs and boll injury in cotton, Gossypium hirsutum L., and their spatial association with a multispectral vegetation index (normalized difference vegetation index [NDVI]). Using the spatial analysis by distance indices analyses, stink bugs were less frequently aggregated (17% for adults and 4% for nymphs) than boll injury (36%). NDVI values were also significantly aggregated within fields in 19 of 48 analyses (40%), with the majority of significant indices occurring in July and August. Paired NDVI datasets from different sampling dates were frequently associated (86.5% for weekly intervals among datasets). Spatial distributions of both stink bugs and boll injury were less stable than for NDVI, with positive associations varying from 12.5 to 25% for adult stink bugs for weekly intervals, depending on species. Spatial distributions of boll injury from stink bug feeding were more stable than stink bugs, with 46% positive associations among paired datasets with weekly intervals. NDVI values were positively associated with boll injury from stink bug feeding in 11 out of 22 analyses, with no significant negative associations. This indicates that NDVI has potential as a component of site-specific management. Future work should continue to examine the value of remote sensing for insect management in cotton, with an aim to develop tools such as risk assessment maps that will help growers to reduce insecticide inputs. © The Authors 2016. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Specific cross-linking of capsid proteins to virus RNA by ultraviolet irradiation of polio virus

Energy Technology Data Exchange (ETDEWEB)

Wetz, K.; Habermehl, K.O. (Freie Univ. Berlin (Germany, F.R.))

1982-04-01

Poliovirus was irradiated with u.v. light under conditions causing approx. 5% cross-linking of capsid protein to virus RNA. Cross-linked RNA-protein complexes, freed from unbound protein, were treated with nuclease, and then analysed on SDS-polyacrylamide gels. The smallest capsid polypeptide VP4 was found to be associated with the RNA to the greatest degree, followed by VP2 and VP1, while VP3 was attached only in trace amounts. Low radiation doses, which produced cross-linking of RNA to protein, did not cause breakdown of the virus particles or conformational changes of the capsid as examined physically and serologically. However, higher doses caused structural alterations of the virus capsid.

Specific cross-linking of capsid proteins to virus RNA by ultraviolet irradiation of polio virus

International Nuclear Information System (INIS)

Wetz, K.; Habermehl, K.-O.

1982-01-01

Poliovirus was irradiated with u.v. light under conditions causing approx. 5% cross-linking of capsid protein to virus RNA. Cross-linked RNA-protein complexes, freed from unbound protein, were treated with nuclease, and then analysed on SDS-polyacrylamide gels. The smallest capsid polypeptide VP4 was found to be associated with the RNA to the greatest degree, followed by VP2 and VP1, while VP3 was attached only in trace amounts. Low radiation doses, which produced cross-linking of RNA to protein, did not cause breakdown of the virus particles or conformational changes of the capsid as examined physically and serologically. However, higher doses caused structural alterations of the virus capsid. (author)

Hepatitis B virus core protein allosteric modulators can distort and disrupt intact capsids

Science.gov (United States)

Schlicksup, Christopher John; Wang, Joseph Che-Yen; Francis, Samson; Venkatakrishnan, Balasubramanian; Turner, William W; VanNieuwenhze, Michael

2018-01-01

Defining mechanisms of direct-acting antivirals facilitates drug development and our understanding of virus function. Heteroaryldihydropyrimidines (HAPs) inappropriately activate assembly of hepatitis B virus (HBV) core protein (Cp), suppressing formation of virions. We examined a fluorophore-labeled HAP, HAP-TAMRA. HAP-TAMRA induced Cp assembly and also bound pre-assembled capsids. Kinetic and spectroscopic studies imply that HAP-binding sites are usually not available but are bound cooperatively. Using cryo-EM, we observed that HAP-TAMRA asymmetrically deformed capsids, creating a heterogeneous array of sharp angles, flat regions, and outright breaks. To achieve high resolution reconstruction (particle symmetry. We deduced that HAP-TAMRA caused quasi-sixfold vertices to become flatter and fivefold more angular. This transition led to asymmetric faceting. That a disordered crosslink could rescue symmetry implies that capsids have tensegrity properties. Capsid distortion and disruption is a new mechanism by which molecules like the HAPs can block HBV infection. PMID:29377794

The two capsid proteins of maize rayado fino virus contain common peptide sequences.

Science.gov (United States)

Falk, B W; Tsai, J H

1986-01-01

Virions of maize rayado fino virus (MRFV) were purified and two major capsid proteins (ca. Mr 29,000 and 22,000) were resolved by SDS-PAGE. When the two major capsid proteins were isolated from gels and compared by one-dimensional peptide mapping after digestion with Staphylococcus aureus V-8 protease, indistinguishable peptide maps were obtained, suggesting that these two proteins contain common peptide sequences. Some preparations also showed minor protein components that were intermediate between the Mr 22,000 and Mr 29,000 capsid proteins. One of the minor proteins, ca. Mr 27,000, gave a peptide map indistinguishable from the major capsid proteins. In vitro ageing of partially purified preparations or virion treatment with proteolytic enzymes failed to show conversion of the Mr 29,000 protein to a Mr 22,000. Protease inhibitors added to the buffers used for virion purification did not affect the apparent 1:3 ratio of 29,000 to 22,000 proteins in the purified preparations.

Effect of capsid proteins to ICG mass ratio on fluorescent quantum yield of virus-resembling optical nano-materials

Science.gov (United States)

Gupta, Sharad; Ico, Gerardo; Matsumura, Paul; Rao, A. L. N.; Vullev, Valentine; Anvari, Bahman

2012-03-01

We recently reported construction of a new type of optical nano-construct composed of genome-depleted plant infecting brome mosaic virus (BMV) doped with Indocyanine green (ICG), an FDA-approved chromophore. We refer to these constructs as optical viral ghosts (OVGs) since only the capsid protein (CP) subunits of BMV remain to encapsulate ICG. To utilize OVGs as effective nano-probes in fluorescence imaging applications, their fluorescence quantum yield needs to be maximized. In this study, we investigate the effect of altering the CP to ICG mass ratio on the fluorescent quantum yield of OVGs. Results of this study provide the basis for construction of OVGs with optimal amounts of CP and ICG to yield maximal fluorescence quantum yield.

Harvesting Venom Toxins from Assassin Bugs and Other Heteropteran Insects.

Science.gov (United States)

Walker, Andrew Allan; Rosenthal, Max; Undheim, Eivind E A; King, Glenn F

2018-04-21

Heteropteran insects such as assassin bugs (Reduviidae) and giant water bugs (Belostomatidae) descended from a common predaceous and venomous ancestor, and the majority of extant heteropterans retain this trophic strategy. Some heteropterans have transitioned to feeding on vertebrate blood (such as the kissing bugs, Triatominae; and bed bugs, Cimicidae) while others have reverted to feeding on plants (most Pentatomomorpha). However, with the exception of saliva used by kissing bugs to facilitate blood-feeding, little is known about heteropteran venoms compared to the venoms of spiders, scorpions and snakes. One obstacle to the characterization of heteropteran venom toxins is the structure and function of the venom/labial glands, which are both morphologically complex and perform multiple biological roles (defense, prey capture, and extra-oral digestion). In this article, we describe three methods we have successfully used to collect heteropteran venoms. First, we present electrostimulation as a convenient way to collect venom that is often lethal when injected into prey animals, and which obviates contamination by glandular tissue. Second, we show that gentle harassment of animals is sufficient to produce venom extrusion from the proboscis and/or venom spitting in some groups of heteropterans. Third, we describe methods to harvest venom toxins by dissection of anaesthetized animals to obtain the venom glands. This method is complementary to other methods, as it may allow harvesting of toxins from taxa in which electrostimulation and harassment are ineffective. These protocols will enable researchers to harvest toxins from heteropteran insects for structure-function characterization and possible applications in medicine and agriculture.

Structure of a Human Astrovirus Capsid-Antibody Complex and Mechanistic Insights into Virus Neutralization

Energy Technology Data Exchange (ETDEWEB)

Bogdanoff, Walter A.; Campos, Jocelyn; Perez, Edmundo I.; Yin, Lu; Alexander, David L.; DuBois, Rebecca M. (UCSC)

2016-11-02

Human astroviruses (HAstVs) are a leading cause of viral diarrhea in young children, the immunocompromised, and the elderly. There are no vaccines or antiviral therapies against HAstV disease. Several lines of evidence point to the presence of protective antibodies in healthy adults as a mechanism governing protection against reinfection by HAstV. However, development of anti-HAstV therapies is hampered by the gap in knowledge of protective antibody epitopes on the HAstV capsid surface. Here, we report the structure of the HAstV capsid spike domain bound to the neutralizing monoclonal antibody PL-2. The antibody uses all six complementarity-determining regions to bind to a quaternary epitope on each side of the dimeric capsid spike. We provide evidence that the HAstV capsid spike is a receptor-binding domain and that the antibody neutralizes HAstV by blocking virus attachment to cells. We identify patches of conserved amino acids that overlap the antibody epitope and may comprise a receptor-binding site. Our studies provide a foundation for the development of therapies to prevent and treat HAstV diarrheal disease.

IMPORTANCEHuman astroviruses (HAstVs) infect nearly every person in the world during childhood and cause diarrhea, vomiting, and fever. Despite the prevalence of this virus, little is known about how antibodies in healthy adults protect them against reinfection. Here, we determined the crystal structure of a complex of the HAstV capsid protein and a virus-neutralizing antibody. We show that the antibody binds to the outermost spike domain of the capsid, and we provide evidence that the antibody blocks virus attachment to human cells. Importantly, our findings suggest that a subunit-based vaccine focusing the immune system on the HAstV capsid spike domain could be effective in protecting children against HAstV disease.

SCHEMA computational design of virus capsid chimeras: calibrating how genome packaging, protection, and transduction correlate with calculated structural disruption.

Science.gov (United States)

Ho, Michelle L; Adler, Benjamin A; Torre, Michael L; Silberg, Jonathan J; Suh, Junghae

2013-12-20

Adeno-associated virus (AAV) recombination can result in chimeric capsid protein subunits whose ability to assemble into an oligomeric capsid, package a genome, and transduce cells depends on the inheritance of sequence from different AAV parents. To develop quantitative design principles for guiding site-directed recombination of AAV capsids, we have examined how capsid structural perturbations predicted by the SCHEMA algorithm correlate with experimental measurements of disruption in seventeen chimeric capsid proteins. In our small chimera population, created by recombining AAV serotypes 2 and 4, we found that protection of viral genomes and cellular transduction were inversely related to calculated disruption of the capsid structure. Interestingly, however, we did not observe a correlation between genome packaging and calculated structural disruption; a majority of the chimeric capsid proteins formed at least partially assembled capsids and more than half packaged genomes, including those with the highest SCHEMA disruption. These results suggest that the sequence space accessed by recombination of divergent AAV serotypes is rich in capsid chimeras that assemble into 60-mer capsids and package viral genomes. Overall, the SCHEMA algorithm may be useful for delineating quantitative design principles to guide the creation of libraries enriched in genome-protecting virus nanoparticles that can effectively transduce cells. Such improvements to the virus design process may help advance not only gene therapy applications but also other bionanotechnologies dependent upon the development of viruses with new sequences and functions.

Bug22 influences cilium morphology and the post-translational modification of ciliary microtubules

Directory of Open Access Journals (Sweden)

Teresa Mendes Maia

2014-01-01

Cilia and flagella are organelles essential for motility and sensing of environmental stimuli. Depending on the cell type, cilia acquire a defined set of functions and, accordingly, are built with an appropriate length and molecular composition. Several ciliary proteins display a high degree of conservation throughout evolution and mutations in ciliary genes are associated with various diseases such as ciliopathies and infertility. Here, we describe the role of the highly conserved ciliary protein, Bug22, in Drosophila. Previous studies in unicellular organisms have shown that Bug22 is required for proper cilia function, but its exact role in ciliogenesis has not been investigated yet. Null Bug22 mutant flies display cilia-associated phenotypes and nervous system defects. Furthermore, sperm differentiation is blocked at the individualization stage, due to impaired migration of the individualization machinery. Tubulin post-translational modifications (PTMs such as polyglycylation, polyglutamylation or acetylation, are determinants of microtubule (MT functions and stability in centrioles, cilia and neurons. We found defects in the timely incorporation of polyglycylation in sperm axonemal MTs of Bug22 mutants. In addition, we found that depletion of human Bug22 in RPE1 cells resulted in the appearance of longer cilia and reduced axonemal polyglutamylation. Our work identifies Bug22 as a protein that plays a conserved role in the regulation of PTMs of the ciliary axoneme.

Bug Distribution and Statistical Pattern Classification.

Science.gov (United States)

Tatsuoka, Kikumi K.; Tatsuoka, Maurice M.

1987-01-01

The rule space model permits measurement of cognitive skill acquisition and error diagnosis. Further discussion introduces Bayesian hypothesis testing and bug distribution. An illustration involves an artificial intelligence approach to testing fractions and arithmetic. (Author/GDC)

Characterization of the mode of action of a potent dengue virus capsid inhibitor.

Science.gov (United States)

Scaturro, Pietro; Trist, Iuni Margaret Laura; Paul, David; Kumar, Anil; Acosta, Eliana G; Byrd, Chelsea M; Jordan, Robert; Brancale, Andrea; Bartenschlager, Ralf

2014-10-01

Dengue viruses (DV) represent a significant global health burden, with up to 400 million infections every year and around 500,000 infected individuals developing life-threatening disease. In spite of attempts to develop vaccine candidates and antiviral drugs, there is a lack of approved therapeutics for the treatment of DV infection. We have previously reported the identification of ST-148, a small-molecule inhibitor exhibiting broad and potent antiviral activity against DV in vitro and in vivo (C. M. Byrd et al., Antimicrob. Agents Chemother. 57:15-25, 2013, doi:10 .1128/AAC.01429-12). In the present study, we investigated the mode of action of this promising compound by using a combination of biochemical, virological, and imaging-based techniques. We confirmed that ST-148 targets the capsid protein and obtained evidence of bimodal antiviral activity affecting both assembly/release and entry of infectious DV particles. Importantly, by using a robust bioluminescence resonance energy transfer-based assay, we observed an ST-148-dependent increase of capsid self-interaction. These results were corroborated by molecular modeling studies that also revealed a plausible model for compound binding to capsid protein and inhibition by a distinct resistance mutation. These results suggest that ST-148-enhanced capsid protein self-interaction perturbs assembly and disassembly of DV nucleocapsids, probably by inducing structural rigidity. Thus, as previously reported for other enveloped viruses, stabilization of capsid protein structure is an attractive therapeutic concept that also is applicable to flaviviruses. Dengue viruses are arthropod-borne viruses representing a significant global health burden. They infect up to 400 million people and are endemic to subtropical and tropical areas of the world. Currently, there are neither vaccines nor approved therapeutics for the prophylaxis or treatment of DV infections, respectively. This study reports the characterization of the

Efficacy of an Essential Oil-Based Pesticide for Controlling Bed Bug (Cimex lectularius Infestations in Apartment Buildings

Directory of Open Access Journals (Sweden)

Changlu Wang

2014-11-01

Full Text Available Bed bugs (Cimex lectularius L. and Cimex hemipterus F. are among the most difficult urban pests to manage. Many essential oil-based bed bug control products that are considered reduced risk to mammals compared to synthetic insect neurotoxins have become commercially available, but their effectiveness as a stand-alone control method is unknown. This study assessed the field efficacy of an essential oil-based bed bug control product (EcoRaider; a.i. 1% geraniol + 1% cedar oil + 2% sodium lauryl sulfate compared to a pyrethroid and neonicotinoid mixture spray (0.075% Temprid SC; a.i. beta-cyfluthrin + imidacloprid. After 12 weeks, the three treatments—EcoRaider, Temprid SC, and EcoRaider + Temprid SC caused 92.5 ± 2.7, 92.9 ± 3.0, and 91.7% ± 2.7% bed bug count reduction, respectively. No significant differences existed in the bed bug reduction among the treatments. Bed bugs were eliminated from only 22% of the treated apartments. Among those still with bed bugs, 76% of the residents did not know bed bugs were present. We documented the residents’ self-control practices and discussed the potential of using essential oil-based insecticides in bed bug management programs to minimize the health risks to building occupants and pets and to slow down the development of insecticide resistance.

Comparing Fine-Grained Source Code Changes And Code Churn For Bug Prediction

NARCIS (Netherlands)

Giger, E.; Pinzger, M.; Gall, H.C.

2011-01-01

A significant amount of research effort has been dedicated to learning prediction models that allow project managers to efficiently allocate resources to those parts of a software system that most likely are bug-prone and therefore critical. Prominent measures for building bug prediction models are

Usage of data warehouse for analysing software's bugs

Science.gov (United States)

Živanov, Danijel; Krstićev, Danijela Boberić; Mirković, Duško

2017-07-01

We analysed the database schema of Bugzilla system and taking into account user's requirements for reporting, we presented a dimensional model for the data warehouse which will be used for reporting software defects. The idea proposed in this paper is not to throw away Bugzilla system because it certainly has many strengths, but to make integration of Bugzilla and the proposed data warehouse. Bugzilla would continue to be used for recording bugs that occur during the development and maintenance of software while the data warehouse would be used for storing data on bugs in an appropriate form, which is more suitable for analysis.

Insecticide resistance and resistance mechanisms in bed bugs, Cimex spp. (Hemiptera: Cimicidae).

Science.gov (United States)

Dang, Kai; Doggett, Stephen L; Veera Singham, G; Lee, Chow-Yang

2017-06-29

The worldwide resurgence of bed bugs [both Cimex lectularius L. and Cimex hemipterus (F.)] over the past two decades is believed in large part to be due to the development of insecticide resistance. The transcriptomic and genomic studies since 2010, as well as morphological, biochemical and behavioral studies, have helped insecticide resistance research on bed bugs. Multiple resistance mechanisms, including penetration resistance through thickening or remodelling of the cuticle, metabolic resistance by increased activities of detoxification enzymes (e.g. cytochrome P450 monooxygenases and esterases), and knockdown resistance by kdr mutations, have been experimentally identified as conferring insecticide resistance in bed bugs. Other candidate resistance mechanisms, including behavioral resistance, some types of physiological resistance (e.g. increasing activities of esterases by point mutations, glutathione S-transferase, target site insensitivity including altered AChEs, GABA receptor insensitivity and altered nAChRs), symbiont-mediated resistance and other potential, yet undiscovered mechanisms may exist. This article reviews recent studies of resistance mechanisms and the genes governing insecticide resistance, potential candidate resistance mechanisms, and methods of monitoring insecticide resistance in bed bugs. This article provides an insight into the knowledge essential for the development of both insecticide resistance management (IRM) and integrated pest management (IPM) strategies for successful bed bug management.

Use of artificial substrates of different colors for oviposition by the brown stink bug Euschistus heros (Hemiptera, Pentatomidae

Directory of Open Access Journals (Sweden)

Diones Krinski

2013-09-01

Full Text Available Aiming to contribute to a rearing methodology for the brown stink bug, Euschistus heros, in the laboratory, we evaluated oviposition on artificial substrates of different colors. During six days, oviposition was evaluated daily, by counting the total number of eggs, number of clutches, and eggs/clutch. Females laid 12,463 eggs, in 1,677 clutches, resulting in an average of 7.28 ± 0.44 eggs/clutch. Black, brown, and green felt had the most eggs and clutches. The results demonstrated that many colors are suitable as oviposition substrate for E. heros, providing information for the mass rearing of this insect.

Do-it-yourself Bed Bug Control

Science.gov (United States)

... Contact Us Share Do-it-yourself Bed Bug Control Información relacionada disponible en español Can you treat ... all of the residents to participate. Achieving complete control can take weeks to months, depending on the ...

Bed Bugs are Public Health Pests

Science.gov (United States)

EPA and the Centers for Disease Control and Prevention (CDC) issued a joint statement on the public health impacts of bed bugs, which are blood-sucking ectoparasites (external parasites). EPA also has a pesticide registration notice on this topic.

Functional dissection of the alphavirus capsid protease: sequence requirements for activity.

Science.gov (United States)

Thomas, Saijo; Rai, Jagdish; John, Lijo; Günther, Stephan; Drosten, Christian; Pützer, Brigitte M; Schaefer, Stephan

2010-11-18

The alphavirus capsid is multifunctional and plays a key role in the viral life cycle. The nucleocapsid domain is released by the self-cleavage activity of the serine protease domain within the capsid. All alphaviruses analyzed to date show this autocatalytic cleavage. Here we have analyzed the sequence requirements for the cleavage activity of Chikungunya virus capsid protease of genus alphavirus. Amongst alphaviruses, the C-terminal amino acid tryptophan (W261) is conserved and found to be important for the cleavage. Mutating tryptophan to alanine (W261A) completely inactivated the protease. Other amino acids near W261 were not having any effect on the activity of this protease. However, serine protease inhibitor AEBSF did not inhibit the activity. Through error-prone PCR we found that isoleucine 227 is important for the effective activity. The loss of activity was analyzed further by molecular modelling and comparison of WT and mutant structures. It was found that lysine introduced at position 227 is spatially very close to the catalytic triad and may disrupt electrostatic interactions in the catalytic site and thus inactivate the enzyme. We are also examining other sequence requirements for this protease activity. We analyzed various amino acid sequence requirements for the activity of ChikV capsid protease and found that amino acids outside the catalytic triads are important for the activity.

Integrating Symbolic Execution with Sensornet Simulation for Efficient Bug Finding

OpenAIRE

Österlind, Fredrik; Sasnauskas, Raimondas; Dustmann, Oscar Soria; Dunkels, Adam; Wehrle, Klaus

2010-01-01

High-coverage testing of sensornet applications is vital for pre-deployment bug cleansing, but has previously been difficult due to the limited set of available tools. We integrate the KleeNet symbolic execution engine with the COOJA network simulator to allow for straight-forward and intuitive high-coverage testing initiated from a simulation environment. A tight coupling of simulation and testing helps detect, narrow down, and fix complex interaction bugs in an early ...

Structural Transitions and Energy Landscape for Cowpea Chlorotic Mottle Virus Capsid Mechanics from Nanomanipulation in Vitro and in Silico

Science.gov (United States)

Kononova, Olga; Snijder, Joost; Brasch, Melanie; Cornelissen, Jeroen; Dima, Ruxandra I.; Marx, Kenneth A.; Wuite, Gijs J. L.; Roos, Wouter H.; Barsegov, Valeri

2013-10-01

Physical properties of capsids of plant and animal viruses are important factors in capsid self-assembly, survival of viruses in the extracellular environment, and their cell infectivity. Virus shells can have applications as nanocontainers and delivery vehicles in biotechnology and medicine. Combined AFM experiments and computational modeling on sub-second timescales of the indentation nanomechanics of Cowpea Chlorotic Mottle Virus (CCMV) capsid show that the capsid's physical properties are dynamic and local characteristics of the structure, which depend on the magnitude and geometry of mechanical input. Surprisingly, under large deformations the CCMV capsid transitions to the collapsed state without substantial local structural alterations. The enthalpy change in this deformation state dH = 11.5 - 12.8 MJ/mol is mostly due to large-amplitude out-of-plane excitations, which contribute to the capsid bending, and the entropy change TdS = 5.1 - 5.8 MJ/mol is mostly due to coherent in-plane rearrangements of protein chains, which result in the capsid stiffening. Dynamic coupling of these modes defines the extent of elasticity and reversibility of capsid mechanical deformation. This emerging picture illuminates how unique physico-chemical properties of protein nanoshells help define their structure and morphology, and determine their viruses' biological function.

Comparative Performance Analysis of Machine Learning Techniques for Software Bug Detection

OpenAIRE

Saiqa Aleem; Luiz Fernando Capretz; Faheem Ahmed

2015-01-01

Machine learning techniques can be used to analyse data from different perspectives and enable developers to retrieve useful information. Machine learning techniques are proven to be useful in terms of software bug prediction. In this paper, a comparative performance analysis of different machine learning techniques is explored f or software bug prediction on public available data sets. Results showed most of the mac ...

Effectiveness of a Reduced-Risk Insecticide Based Bed Bug Management Program in Low-Income Housing

Directory of Open Access Journals (Sweden)

Narinderpal Singh

2013-11-01

Full Text Available Bed bug (Cimex lectularius L. infestations are becoming increasingly common in low-income communities. Once they are introduced, elimination is very difficult. As part of the efforts to develop effective and safe bed bug management programs, we conducted a laboratory study evaluating the efficacy of a reduced-risk insecticide—Alpine aerosol (0.5% dinotefuran. We then conducted a field evaluation of a reduced-risk insecticide based integrated pest management (IPM program in low-income family apartments with young children. In laboratory evaluations, direct spray and 5 min exposure to dry Alpine aerosol residue caused 100.0 ± 0.0 and 91.7 ± 8.3% mortality to bed bug nymphs, respectively. Direct Alpine aerosol spray killed 91.3 ± 4.3% of the eggs. The IPM program included education, steam, bagging infested linens, placing intercepting devices under furniture legs and corners of rooms, applying Alpine aerosol and Alpine dust (0.25% dinotefuran, 95% diatomaceous earth dust, and regularly scheduled monitoring and re-treatment. Nine apartments ranging from 1–1,428 (median: 29 bed bugs based on visual inspection and Climbup interceptor counts were included. Over a 6-month period, an average 172 g insecticide (Alpine aerosol + Alpine dust was used in each apartment, a 96% reduction in pesticide usage compared to chemical only treatment reported in a similar environment. The IPM program resulted in an average of 96.8 ± 2.2% reduction in the number of bed bugs. However, elimination of bed bugs was only achieved in three lightly infested apartments (<30 bed bugs at the beginning. Elimination success was closely correlated with the level of bed bug populations.

Facilitating the use of alternative capsid control methods towards sustainable production of organic cocoa in Ghana

OpenAIRE

Ayenor, G.K.; Huis, van, A.; Obeng-Ofori, D.; Padi, B.; Röling, N.G.

2007-01-01

Cocoa (Theobroma cacao L.) is an important foreign exchange earner for Ghana. However, production is constrained by a high incidence of pests and diseases. Based on farmers' needs, this study focused on the control of capsids, mainly Sahlbergella singularis Haglund and Distantiella theobroma (Distant) (both Hemiptera: Miridae). Annual crop loss caused by capsids is estimated at 25¿30%. To control capsids, formal research recommends application of synthetic insecticides four times between Augu...

Area-wide management approach for tarnished plant bug in the Mississippi Delta

Science.gov (United States)

The tarnished plant bug, Lygus lineolaris (Palisot de Beauvois), is the major insect pest of cotton, Gossypium hirsutum (L.), within the Mid-South region. From 2001 to 2012, the tarnished plant bug has been the number one insect pest of cotton in Louisiana and Mississippi in eleven and nine of those...

ASSESSMENT OF HEAVY METALS CONTENTS IN BOTTOM SEDIMENTS OF BUG RIVER

Directory of Open Access Journals (Sweden)

Elżbieta Skorbiłowicz

2014-07-01

Full Text Available The development of industry, agriculture, and transport contributes to an increased environmental pollution by heavy metals. The aim of the study was preliminary assessment of the contents of selected metals (lead, cobalt, copper, chromium, cadmium and nickel in the sediments of Bug river. The study comprised part of the river flowing through Poland. It was found that the Bug river sediments are not contaminated in respect to the content of tested metals. Based on the analysis of the study results, these metals can be lined up in the following order: Cr > Pb > Cu > Ni > Co > Cd. Statistical analysis showed that copper and chromium occur in Bug river sediments in forms bindings with organic matter in majority of cases. The granulometric analysis of sediments from Bug river revealed the largest percentage of two fractions: 1.0–0.2 mm with average of 47.7 ± 19.77% and 0.2–0.1 mm with average of 20.6 ± 7.7%. These are the dominant fractions with the accumulation of metals in river sediments, which has been confirmed by statistical analysis.

Intra- and inter-subunit disulfide bond formation is nonessential in adeno-associated viral capsids.

Directory of Open Access Journals (Sweden)

Nagesh Pulicherla

Full Text Available The capsid proteins of adeno-associated viruses (AAV have five conserved cysteine residues. Structural analysis of AAV serotype 2 reveals that Cys289 and Cys361 are located adjacent to each other within each monomer, while Cys230 and Cys394 are located on opposite edges of each subunit and juxtaposed at the pentamer interface. The Cys482 residue is located at the base of a surface loop within the trimer region. Although plausible based on molecular dynamics simulations, intra- or inter-subunit disulfides have not been observed in structural studies. In the current study, we generated a panel of Cys-to-Ser mutants to interrogate the potential for disulfide bond formation in AAV capsids. The C289S, C361S and C482S mutants were similar to wild type AAV with regard to titer and transduction efficiency. However, AAV capsid protein subunits with C230S or C394S mutations were prone to proteasomal degradation within the host cells. Proteasomal inhibition partially blocked degradation of mutant capsid proteins, but failed to rescue infectious virions. While these results suggest that the Cys230/394 pair is critical, a C394V mutant was found viable, but not the corresponding C230V mutant. Although the exact nature of the structural contribution(s of Cys230 and Cys394 residues to AAV capsid formation remains to be determined, these results support the notion that disulfide bond formation within the Cys289/361 or Cys230/394 pair appears to be nonessential. These studies represent an important step towards understanding the role of inter-subunit interactions that drive AAV capsid assembly.

Creating a marketplace for green roofs in Chicago

International Nuclear Information System (INIS)

Vitt Sale, L.; Berkshire, M.

2004-01-01

Since 2003, the Chicago Department of Planning and Development has been encouraging city developers to consider installing green roofs on buildings in Chicago, with the belief that this practice results in mitigation of the urban heat island effect, cleaner runoff leaving green roofs, sound attenuation, aesthetic value, oxygen production, and mitigation of carbon dioxide emissions. However, the benefits to developers, which include reduced stormwater runoff, extended roof life and energy savings, in total do not offset the first cost premium of a green roof. Despite this, and with no mandate requiring green roofs, the marketplace is growing. After seeing green roofs on a tour in Europe, the mayor of Chicago encouraged the first design and installation of a 20,300 square foot demonstration green roof in Chicago, and other city-sponsored pilot projects followed shortly after. Since then, the number of green roofs in Chicago has grown to over one million square feet. A map of Chicago showing locations of most of the projects was presented. It was suggested that lower prices for green roofs, higher energy costs and an inclination to invest in long-term strategies would accelerate the market. In an effort to engage the public in dialogue, the Department of Planning and Development held seminars to promote the benefits of green roofs . Participants had many questions about the applicability of green roofs to Chicago, expressing skepticism that Chicago's climate would provide the same benefits as in Europe. Other concerns were expressed regarding the devaluation of property values resulting from placing green roofs on buildings; doubts about roof leaks; maintenance practices; and, bugs and mold. Since the first cost premium of the system remains a question, most participants expressed interest in some kind of incentive program, but remained open-minded if benefits could be proved. 6 figs

Creating a marketplace for green roofs in Chicago

Energy Technology Data Exchange (ETDEWEB)

Vitt Sale, L. [Wright and Co. Chicago, IL (United States); Berkshire, M. [City of Chicago, IL (United States)

2004-07-01

Since 2003, the Chicago Department of Planning and Development has been encouraging city developers to consider installing green roofs on buildings in Chicago, with the belief that this practice results in mitigation of the urban heat island effect, cleaner runoff leaving green roofs, sound attenuation, aesthetic value, oxygen production, and mitigation of carbon dioxide emissions. However, the benefits to developers, which include reduced stormwater runoff, extended roof life and energy savings, in total do not offset the first cost premium of a green roof. Despite this, and with no mandate requiring green roofs, the marketplace is growing. After seeing green roofs on a tour in Europe, the mayor of Chicago encouraged the first design and installation of a 20,300 square foot demonstration green roof in Chicago, and other city-sponsored pilot projects followed shortly after. Since then, the number of green roofs in Chicago has grown to over one million square feet. A map of Chicago showing locations of most of the projects was presented. It was suggested that lower prices for green roofs, higher energy costs and an inclination to invest in long-term strategies would accelerate the market. In an effort to engage the public in dialogue, the Department of Planning and Development held seminars to promote the benefits of green roofs . Participants had many questions about the applicability of green roofs to Chicago, expressing skepticism that Chicago's climate would provide the same benefits as in Europe. Other concerns were expressed regarding the devaluation of property values resulting from placing green roofs on buildings; doubts about roof leaks; maintenance practices; and, bugs and mold. Since the first cost premium of the system remains a question, most participants expressed interest in some kind of incentive program, but remained open-minded if benefits could be proved. 6 figs.

Human Cytomegalovirus Nuclear Capsids Associate with the Core Nuclear Egress Complex and the Viral Protein Kinase pUL97.

Science.gov (United States)

Milbradt, Jens; Sonntag, Eric; Wagner, Sabrina; Strojan, Hanife; Wangen, Christina; Lenac Rovis, Tihana; Lisnic, Berislav; Jonjic, Stipan; Sticht, Heinrich; Britt, William J; Schlötzer-Schrehardt, Ursula; Marschall, Manfred

2018-01-13

The nuclear phase of herpesvirus replication is regulated through the formation of regulatory multi-component protein complexes. Viral genomic replication is followed by nuclear capsid assembly, DNA encapsidation and nuclear egress. The latter has been studied intensely pointing to the formation of a viral core nuclear egress complex (NEC) that recruits a multimeric assembly of viral and cellular factors for the reorganization of the nuclear envelope. To date, the mechanism of the association of human cytomegalovirus (HCMV) capsids with the NEC, which in turn initiates the specific steps of nuclear capsid budding, remains undefined. Here, we provide electron microscopy-based data demonstrating the association of both nuclear capsids and NEC proteins at nuclear lamina budding sites. Specifically, immunogold labelling of the core NEC constituent pUL53 and NEC-associated viral kinase pUL97 suggested an intranuclear NEC-capsid interaction. Staining patterns with phospho-specific lamin A/C antibodies are compatible with earlier postulates of targeted capsid egress at lamina-depleted areas. Important data were provided by co-immunoprecipitation and in vitro kinase analyses using lysates from HCMV-infected cells, nuclear fractions, or infectious virions. Data strongly suggest that nuclear capsids interact with pUL53 and pUL97. Combined, the findings support a refined concept of HCMV nuclear trafficking and NEC-capsid interaction.

Human Cytomegalovirus Nuclear Capsids Associate with the Core Nuclear Egress Complex and the Viral Protein Kinase pUL97

Directory of Open Access Journals (Sweden)

Jens Milbradt

2018-01-01

Full Text Available The nuclear phase of herpesvirus replication is regulated through the formation of regulatory multi-component protein complexes. Viral genomic replication is followed by nuclear capsid assembly, DNA encapsidation and nuclear egress. The latter has been studied intensely pointing to the formation of a viral core nuclear egress complex (NEC that recruits a multimeric assembly of viral and cellular factors for the reorganization of the nuclear envelope. To date, the mechanism of the association of human cytomegalovirus (HCMV capsids with the NEC, which in turn initiates the specific steps of nuclear capsid budding, remains undefined. Here, we provide electron microscopy-based data demonstrating the association of both nuclear capsids and NEC proteins at nuclear lamina budding sites. Specifically, immunogold labelling of the core NEC constituent pUL53 and NEC-associated viral kinase pUL97 suggested an intranuclear NEC-capsid interaction. Staining patterns with phospho-specific lamin A/C antibodies are compatible with earlier postulates of targeted capsid egress at lamina-depleted areas. Important data were provided by co-immunoprecipitation and in vitro kinase analyses using lysates from HCMV-infected cells, nuclear fractions, or infectious virions. Data strongly suggest that nuclear capsids interact with pUL53 and pUL97. Combined, the findings support a refined concept of HCMV nuclear trafficking and NEC-capsid interaction.

Bed Bugs Appearance and Life Cycle

Science.gov (United States)

Identify these bugs accurately, from egg to nymph to adult. Adults are about the size of an apple seed, brown and oval-shaped, and either flat or balloon-like depending on how recently fed. Nymphs are a bit smaller and translucent or whitish yellow.

Virus Capsids as Targeted Nanoscale Delivery Vessels of Photoactive Compounds for Site-Specific Photodynamic Therapy

Science.gov (United States)

Cohen, Brian A.

The research presented in this work details the use of a viral capsid as an addressable delivery vessel of photoactive compounds for use in photodynamic therapy. Photodynamic therapy is a treatment that involves the interaction of light with a photosensitizing molecule to create singlet oxygen, a reactive oxygen species. Overproduction of singlet oxygen in cells can cause oxidative damage leading to cytotoxicity and eventually cell death. Challenges with the current generation of FDA-approved photosensitizers for photodynamic therapy primarily stem from their lack of tissue specificity. This work describes the packaging of photoactive cationic porphyrins inside the MS2 bacteriophage capsid, followed by external modification of the capsid with cancer cell-targeting G-quadruplex DNA aptamers to generate a tumor-specific photosensitizing agent. First, a cationic porphyrin is loaded into the capsids via nucleotide-driven packaging, a process that involves charge interaction between the porphyrin and the RNA inside the capsid. Results show that over 250 porphyrin molecules associate with the RNA within each MS2 capsid. Removal of RNA from the capsid severely inhibits the packaging of the cationic porphyrins. Porphyrin-virus constructs were then shown to photogenerate singlet oxygen, and cytotoxicity in non-targeted photodynamic treatment experiments. Next, each porphyrin-loaded capsid is externally modified with approximately 60 targeting DNA aptamers by employing a heterobifunctional crosslinking agent. The targeting aptamer is known to bind the protein nucleolin, a ubiquitous protein that is overexpressed on the cell surface by many cancer cell types. MCF-7 human breast carcinoma cells and MCF-10A human mammary epithelial cells were selected as an in vitro model for breast cancer and normal tissue, respectively. Fluorescently tagged virus-aptamer constructs are shown to selectively target MCF-7 cells versus MCF-10A cells. Finally, results are shown in which porphyrin

Bed Bug Clearinghouse by Type of Resource

Science.gov (United States)

This information is to help states, communities, and consumers prevent and control bed bug infestations. These brochures, fact sheets, manuals, posters, checklists, videos, and more provide guidance such as hotel room inspection and pesticide safety.

Changes in the stability and biomechanics of P22 bacteriophage capsid during maturation.

Science.gov (United States)

Kant, Ravi; Llauró, Aida; Rayaprolu, Vamseedhar; Qazi, Shefah; de Pablo, Pedro J; Douglas, Trevor; Bothner, Brian

2018-03-15

The capsid of P22 bacteriophage undergoes a series of structural transitions during maturation that guide it from spherical to icosahedral morphology. The transitions include the release of scaffold proteins and capsid expansion. Although P22 maturation has been investigated for decades, a unified model that incorporates thermodynamic and biophysical analyses is not available. A general and specific model of icosahedral capsid maturation is of significant interest to theoreticians searching for fundamental principles as well as virologists and material scientists seeking to alter maturation to their advantage. To address this challenge, we have combined the results from orthogonal biophysical techniques including differential scanning fluorimetry, atomic force microscopy, circular dichroism, and hydrogen-deuterium exchange mass spectrometry. By integrating these results from single particle and population measurements, an energy landscape of P22 maturation from procapsid through expanded shell to wiffle ball emerged, highlighting the role of metastable structures and the thermodynamics guiding maturation. The propagation of weak quaternary interactions across symmetric elements of the capsid is a key component for stability in P22. A surprising finding is that the progression to wiffle ball, which lacks pentamers, shows that chemical and thermal stability can be uncoupled from mechanical rigidity, elegantly demonstrating the complexity inherent in capsid protein interactions and the emergent properties that can arise from icosahedral symmetry. On a broader scale, this work demonstrates the power of applying orthogonal biophysical techniques to elucidate assembly mechanisms for supramolecular complexes and provides a framework within which other viral systems can be compared. Copyright © 2018 Elsevier B.V. All rights reserved.

Degree Day Requirements for Kudzu Bug (Hemiptera: Plataspidae), a Pest of Soybeans.

Science.gov (United States)

Grant, Jessica I; Lamp, William O

2018-04-02

Understanding the phenology of a new potential pest is fundamental for the development of a management program. Megacopta cribraria Fabricius (Hemiptera: Plataspidae), kudzu bug, is a pest of soybeans first detected in the United States in 2009 and in Maryland in 2013. We observed the phenology of kudzu bug life stages in Maryland, created a Celsius degree-day (CDD) model for development, and characterized the difference between microhabitat and ambient temperatures of both kudzu, Pueraria montana (Lour.) Merr. (Fabales: Fabaceae) and soybeans, Glycine max (L.) Merrill (Fabales: Fabaceae). In 2014, low population numbers yielded limited resolution from field phenology observations. We observed kudzu bug populations persisting within Maryland; but between 2013 and 2016, populations were low compared to populations in the southeastern United States. Based on the degree-day model, kudzu bug eggs require 80 CDD at a minimum temperature of 14°C to hatch. Nymphs require 545 CDD with a minimum temperature of 16°C for development. The CDD model matches field observations when factoring a biofix date of April 1 and a minimum preoviposition period of 17 d. The model suggests two full generations per year in Maryland. Standard air temperature monitors do not affect model predictions for pest management, as microhabitat temperature differences did not show a clear trend between kudzu and soybeans. Ultimately, producers can predict the timing of kudzu bug life stages with the CDD model for the use of timing management plans in soybean fields.

Tyrosine Mutation in AAV9 Capsid Improves Gene Transfer to the Mouse Lung.

Science.gov (United States)

Martini, Sabrina V; Silva, Adriana L; Ferreira, Debora; Rabelo, Rafael; Ornellas, Felipe M; Gomes, Karina; Rocco, Patricia R M; Petrs-Silva, Hilda; Morales, Marcelo M

2016-01-01

Adeno-associated virus (AAV) vectors are being increasingly used as the vector of choice for in vivo gene delivery and gene therapy for many pulmonary diseases. Recently, it was shown that phosphorylation of surface-exposed tyrosine residues from AAV capsid targets the viral particles for ubiquitination and proteasome-mediated degradation, and mutations of these tyrosine residues lead to highly efficient vector transduction in vitro and in vivo in different organs. In this study, we evaluated the pulmonary transgene expression efficacy of AAV9 vectors containing point mutations in surface-exposed capsid tyrosine residues. Eighteen C57BL/6 mice were randomly assigned into three groups: (1) a control group (CTRL) animals underwent intratracheal (i.t.) instillation of saline, (2) the wild-type AAV9 group (WT-AAV9, 1010 vg), and (3) the tyrosine-mutant Y731F AAV9 group (M-AAV9, 1010 vg), which received (i.t.) self-complementary AAV9 vectors containing the DNA sequence of enhanced green fluorescence protein (eGFP). Four weeks after instillation, lung mechanics, morphometry, tissue cellularity, gene expression, inflammatory cytokines, and growth factor expression were analyzed. No significant differences were observed in lung mechanics and morphometry among the experimental groups. However, the number of polymorphonuclear cells was higher in the WT-AAV9 group than in the CTRL and M-AAV9 groups, suggesting that the administration of tyrosine-mutant AAV9 vectors was better tolerated. Tyrosine-mutant AAV9 vectors significantly improved transgene delivery to the lung (30%) compared with their wild-type counterparts, without eliciting an inflammatory response. Our results provide the impetus for further studies to exploit the use of AAV9 vectors as a tool for pulmonary gene therapy. © 2016 The Author(s) Published by S. Karger AG, Basel.

Stress Tolerance of Bed Bugs: A Review of Factors That Cause Trauma to Cimex lectularius and C. Hemipterus

Directory of Open Access Journals (Sweden)

Joshua B. Benoit

2011-04-01

Full Text Available Recent emergence of bed bugs (Cimex spp. has prompted a significant expansion of research devoted to this pest. The ability to survive and recover from stress has significant implications on the distribution and survival of insects, and bed bugs are no exception. Research on bed bug stress tolerance has shown considerable progress and necessitates a review on this topic. Bed bugs have an extraordinary ability to resist dehydration between bloodmeals, and this represents a critical factor allowing their prolonged survival when no host is available. High relative humidities are detrimental to bed bugs, leading to reduced survival in comparison to those held at lower relative humidities. Continual exposure of bed bugs, eggs and mobile stages, to temperatures below freezing and short term exposure (=1 h to temperatures below −16 to −18 °C results in mortality. The upper thermal limit for short term exposure of eggs, nymphs and adults is between 40–45 °C for the common (Cimex lectularius and tropical (C. hemipterus bed bugs. Long-term exposure to temperatures above 35 °C results in significant reduction in survival of mobile bed bugs. Eggs for C. lectularius and C. hemipterus are no longer viable when held below 10 °C or above 37 °C throughout embryogenesis. Blood feeding, although necessary for survival and reproduction, is discussed as a stress due to thermal and osmotic fluctuations that result from ingesting a warm bloodmeal from a vertebrate host. Cold, heat, water stress and blood feeding prompted the expression of heat shock proteins (Hsps. Pesticide application is a common human-induced stress for urban pests, and recent studies have documented pesticide resistance in many bed bug populations. High levels of traumatic insemination (mating of bed bugs has been linked to reduced survival and fecundity along with possibly exposing individuals to microbial infections after cuticular penetration by the paramere (=male reproductive organ

Real-Time Measurement of Volatile Chemicals Released by Bed Bugs during Mating Activities

DEFF Research Database (Denmark)

Kilpinen, Ole Østerlund; Liu, Dezhao; Adamsen, Anders Peter

2012-01-01

In recent years, bed bug (Hemiptera: Cimicidae) problems have increased dramatically in many parts of the world, leading to a renewed interest in their chemical ecology. Most studies of bed bug semiochemicals have been based on the collection of volatiles over a period of time followed by chemical...

Porcine circovirus-2 capsid protein induces cell death in PK15 cells

Energy Technology Data Exchange (ETDEWEB)

Walia, Rupali; Dardari, Rkia, E-mail: rdardari@ucalgary.ca; Chaiyakul, Mark; Czub, Markus

2014-11-15

Studies have shown that Porcine circovirus (PCV)-2 induces apoptosis in PK15 cells. Here we report that cell death is induced in PCV2b-infected PK15 cells that express Capsid (Cap) protein and this effect is enhanced in interferon gamma (IFN-γ)-treated cells. We further show that transient PCV2a and 2b-Cap protein expression induces cell death in PK15 cells at rate similar to PCV2 infection, regardless of Cap protein localization. These data suggest that Cap protein may have the capacity to trigger different signaling pathways involved in cell death. Although further investigation is needed to gain deeper insights into the nature of the pathways involved in Cap-induced cell death, this study provides evidence that PCV2-induced cell death in kidney epithelial PK15 cells can be mapped to the Cap protein and establishes the need for future research regarding the role of Cap-induced cell death in PCV2 pathogenesis. - Highlights: • IFN-γ enhances PCV2 replication that leads to cell death in PK15 cells. • IFN-γ enhances nuclear localization of the PCV2 Capsid protein. • Transient PCV2a and 2b-Capsid protein expression induces cell death. • Cell death is not dictated by specific Capsid protein sub-localization.

Bringing Up Girls in Science (BUGS): The Effectiveness of an Afterschool Environmental Science Program for Increasing Female Students' Interest in Science Careers

Science.gov (United States)

Tyler-Wood, Tandra; Ellison, Amber; Lim, Okyoung; Periathiruvadi, Sita

2012-02-01

Bringing Up Girls in Science (BUGS) was an afterschool program for 4th and 5th grade girls that provided authentic learning experiences in environmental science as well as valuable female mentoring opportunities in an effort to increase participants' academic achievement in science. BUGS participants demonstrated significantly greater amounts of gain in science knowledge as measured by the Iowa Test of Basic Skills in Science (ITBS-S). The original BUGS participants and contrasts have now completed high school and entered college, allowing researchers to assess the long-term impact of the BUGS program. Fourteen former BUGS participants completed two instruments to assess their perceptions of science and science, technology, engineering, and mathematics (STEM) careers. Their results were compared to four contrast groups composed entirely of females: 12 former BUGS contrasts, 10 college science majors, 10 non-science majors, and 9 current STEM professionals. Results indicate that BUGS participants have higher perceptions of science careers than BUGS contrasts. There were no significant differences between BUGS participants, Science Majors, and STEM professionals in their perceptions of science and STEM careers, whereas the BUGS contrast group was significantly lower than BUGS participants, Science Majors, and STEM Professionals. Additional results and implications are discussed within.

Association of Verde plant bug, Creontiades signatus (Hemiptera: Miridae), with cotton boll rot

Science.gov (United States)

Cotton along the Gulf Coast of south Texas has experienced loss from cotton boll rot especially during the last 10 to 15 years, and stink bugs and plant bugs (Hemiptera: Pentatomidae and Miridae) that feed on cotton bolls have been suspected in introducing the disease. A replicated grower field surv...

Interactions among Carbon Dioxide, Heat, and Chemical Lures in Attracting the Bed Bug, Cimex lectularius L. (Hemiptera: Cimicidae

Directory of Open Access Journals (Sweden)

Narinderpal Singh

2012-01-01

Full Text Available Commercial bed bug (Cimex lectularius L. monitors incorporating carbon dioxide (CO2, heat, and chemical lures are being used for detecting bed bugs; however, there are few reported studies on the effectiveness of chemical lures in bed bug monitors and the interactions among chemical lure, CO2, and heat. We screened 12 chemicals for their attraction to bed bugs and evaluated interactions among chemical lures, CO2, and heat. The chemical lure mixture consisting of nonanal, 1-octen-3-ol, spearmint oil, and coriander Egyptian oil was found to be most attractive to bed bugs and significantly increased the trap catches in laboratory bioassays. Adding this chemical lure mixture when CO2 was present increased the trap catches compared with traps baited with CO2 alone, whereas adding heat did not significantly increase trap catches when CO2 was present. Results suggest a combination of chemical lure and CO2 is essential for designing effective bed bug monitors.

Attraction of stink bug (Hemiptera: Pentatomidae) nymphs to Euschistus aggregation pheromone in the field

Science.gov (United States)

Phytophagous stink bugs (Hemiptera: Pentatomidae) are primary pests in most fruit, vegetable, grain, and row crops worldwide. Pheromones have been identified and synthesized for several species of economically important stink bug pests. When yellow pyramid traps are baited with lures containing thes...

Adjacent habitat influence on stink bug (Hemiptera: Pentatomidae) densities and the associated damage at field corn and soybean edges.

Science.gov (United States)

Venugopal, P Dilip; Coffey, Peter L; Dively, Galen P; Lamp, William O

2014-01-01

The local dispersal of polyphagous, mobile insects within agricultural systems impacts pest management. In the mid-Atlantic region of the United States, stink bugs, especially the invasive Halyomorpha halys (Stål 1855), contribute to economic losses across a range of cropping systems. Here, we characterized the density of stink bugs along the field edges of field corn and soybean at different study sites. Specifically, we examined the influence of adjacent managed and natural habitats on the density of stink bugs in corn and soybean fields at different distances along transects from the field edge. We also quantified damage to corn grain, and to soybean pods and seeds, and measured yield in relation to the observed stink bug densities at different distances from field edge. Highest density of stink bugs was limited to the edge of both corn and soybean fields. Fields adjacent to wooded, crop and building habitats harbored higher densities of stink bugs than those adjacent to open habitats. Damage to corn kernels and to soybean pods and seeds increased with stink bug density in plots and was highest at the field edges. Stink bug density was also negatively associated with yield per plant in soybean. The spatial pattern of stink bugs in both corn and soybeans, with significant edge effects, suggests the use of pest management strategies for crop placement in the landscape, as well as spatially targeted pest suppression within fields.

Adjacent habitat influence on stink bug (Hemiptera: Pentatomidae densities and the associated damage at field corn and soybean edges.

Directory of Open Access Journals (Sweden)

P Dilip Venugopal

Full Text Available The local dispersal of polyphagous, mobile insects within agricultural systems impacts pest management. In the mid-Atlantic region of the United States, stink bugs, especially the invasive Halyomorpha halys (Stål 1855, contribute to economic losses across a range of cropping systems. Here, we characterized the density of stink bugs along the field edges of field corn and soybean at different study sites. Specifically, we examined the influence of adjacent managed and natural habitats on the density of stink bugs in corn and soybean fields at different distances along transects from the field edge. We also quantified damage to corn grain, and to soybean pods and seeds, and measured yield in relation to the observed stink bug densities at different distances from field edge. Highest density of stink bugs was limited to the edge of both corn and soybean fields. Fields adjacent to wooded, crop and building habitats harbored higher densities of stink bugs than those adjacent to open habitats. Damage to corn kernels and to soybean pods and seeds increased with stink bug density in plots and was highest at the field edges. Stink bug density was also negatively associated with yield per plant in soybean. The spatial pattern of stink bugs in both corn and soybeans, with significant edge effects, suggests the use of pest management strategies for crop placement in the landscape, as well as spatially targeted pest suppression within fields.

Drosophila Nora virus capsid proteins differ from those of other picorna-like viruses.

Science.gov (United States)

Ekström, Jens-Ola; Habayeb, Mazen S; Srivastava, Vaibhav; Kieselbach, Thomas; Wingsle, Gunnar; Hultmark, Dan

2011-09-01

The recently discovered Nora virus from Drosophila melanogaster is a single-stranded RNA virus. Its published genomic sequence encodes a typical picorna-like cassette of replicative enzymes, but no capsid proteins similar to those in other picorna-like viruses. We have now done additional sequencing at the termini of the viral genome, extending it by 455 nucleotides at the 5' end, but no more coding sequence was found. The completeness of the final 12,333-nucleotide sequence was verified by the production of infectious virus from the cloned genome. To identify the capsid proteins, we purified Nora virus particles and analyzed their proteins by mass spectrometry. Our results show that the capsid is built from three major proteins, VP4A, B and C, encoded in the fourth open reading frame of the viral genome. The viral particles also contain traces of a protein from the third open reading frame, VP3. VP4A and B are not closely related to other picorna-like virus capsid proteins in sequence, but may form similar jelly roll folds. VP4C differs from the others and is predicted to have an essentially α-helical conformation. In a related virus, identified from EST database sequences from Nasonia parasitoid wasps, VP4C is encoded in a separate open reading frame, separated from VP4A and B by a frame-shift. This opens a possibility that VP4C is produced in non-equimolar quantities. Altogether, our results suggest that the Nora virus capsid has a different protein organization compared to the order Picornavirales. Copyright © 2011 Elsevier B.V. All rights reserved.

Bug-in-Ear eCoaching: Impacts on Novice Early Childhood Special Education Teachers

Science.gov (United States)

Grygas Coogle, Christan; Ottley, Jennifer R.; Rahn, Naomi L.; Storie, Sloan

2018-01-01

A multiple-probe, single-case design was used to determine the effects of bug-in-ear eCoaching on teachers' use of two targeted naturalistic communication strategies and focus children's responses to these strategies. Results indicated that bug-in-ear eCoaching enhanced teachers' use of communication strategies and the appropriate responses of…

Importin α1 is required for nuclear import of herpes simplex virus proteins and capsid assembly in fibroblasts and neurons

Science.gov (United States)

Anderson, Fenja; Rother, Franziska; Rudolph, Kathrin; Prank, Ute; Binz, Anne; Hügel, Stefanie; Hartmann, Enno; Bader, Michael; Bauerfeind, Rudolf; Sodeik, Beate

2018-01-01

Herpesviruses are large DNA viruses which depend on many nuclear functions, and therefore on host transport factors to ensure specific nuclear import of viral and host components. While some import cargoes bind directly to certain transport factors, most recruit importin β1 via importin α. We identified importin α1 in a small targeted siRNA screen to be important for herpes simplex virus (HSV-1) gene expression. Production of infectious virions was delayed in the absence of importin α1, but not in cells lacking importin α3 or importin α4. While nuclear targeting of the incoming capsids, of the HSV-1 transcription activator VP16, and of the viral genomes were not affected, the nuclear import of the HSV-1 proteins ICP4 and ICP0, required for efficient viral transcription, and of ICP8 and pUL42, necessary for DNA replication, were reduced. Furthermore, quantitative electron microscopy showed that fibroblasts lacking importin α1 contained overall fewer nuclear capsids, but an increased proportion of mature nuclear capsids indicating that capsid formation and capsid egress into the cytoplasm were impaired. In neurons, importin α1 was also not required for nuclear targeting of incoming capsids, but for nuclear import of ICP4 and for the formation of nuclear capsid assembly compartments. Our data suggest that importin α1 is specifically required for the nuclear localization of several important HSV1 proteins, capsid assembly, and capsid egress into the cytoplasm, and may become rate limiting in situ upon infection at low multiplicity or in terminally differentiated cells such as neurons. PMID:29304174

Milkweed, stink bugs, and Georgia cotton

Science.gov (United States)

In peanut-cotton farmscapes in Georgia, stink bugs, i.e., Nezara viridula (L.)(Say) and Chinavia hilaris (Say), develop in peanut and then disperse at the crop-to-crop interface to feed on fruit in cotton. The main objective of this study was to examine the influence of a habitat of tropical milkwe...

Human Cytomegalovirus pUL47 Modulates Tegumentation and Capsid Accumulation at the Viral Assembly Complex

Science.gov (United States)

Cappadona, Ilaria; Villinger, Clarissa; Schutzius, Gabi; Mertens, Thomas

2015-01-01

ABSTRACT Human cytomegalovirus (HCMV) tegument protein pUL47 is an interaction partner of pUL48 and highly conserved among herpesviruses. It is closely associated with the capsid and has an important function early in infection. Here, we report a specific role of pUL47 in the tegumentation of capsids in the cytoplasm. A newly generated mutant virus (TB-47stop), in which expression of pUL47 is blocked, exhibited a severe impairment in cell-to-cell spread and release of infectivity from infected cells. Ultrastructural analysis of TB-47stop-infected cells clearly showed cytoplasmic accumulations of nonenveloped capsids that were only partially tegumented, indicating that these capsids failed to complete tegumentation. Nevertheless, these accumulations were positive for HCMV inner tegument proteins pp150 and pUL48, suggesting that their attachment to capsids occurs independently of pUL47. Despite these morphological alterations, fully enveloped virus particles were found in the extracellular space and at the viral assembly complex (vAC) of TB-47stop-infected cells, indicating that pUL47 is not essential for the generation of virions. We confirmed findings that incorporation of pUL48 into virions is impaired in the absence of pUL47. Interestingly, pUL47 exhibited a strong nuclear localization in transfected cells, whereas it was found exclusively at the vAC in the context of virus infection. Colocalization of pUL47 and pUL48 at the vAC is consistent with their interaction. We also found a shift to a more nuclear localization of pUL47 when the expression of pUL48 was reduced. Summarizing our results, we hypothesize that pUL48 directs pUL47 to the vAC to promote tegumentation and secondary envelopment of capsids. IMPORTANCE Generation of infectious HCMV particles requires an organized and multistep process involving the action of several viral and cellular proteins as well as protein-protein interactions. A better understanding of these processes is important for

Bug breakfast in the bulletin: human papilloma virus.

Science.gov (United States)

Wallace, Cate; Weisberg, Edith; McCaffery, Kirsten

2007-01-01

The Bug Breakfast topic for October was Human Papilloma Virus (HPV). The presenters covered the epidemiology of HPV, the newly introduced HPV vaccine and social and psychological issues relating to HPV vaccination.

Verde plant bug is associated with cottong boll rot in South Texas cotton

Science.gov (United States)

Verde plant bug was the dominant boll-feeding sucking bug species (>98% of insects collected using a beat bucket) from peak to late bloom in cotton fields near the coast along the Coastal Bend of South Texas, from Port Lavaca to the Lower Rio qrande Valley in 2010 and 2011. It was common in fields w...

Exploring the role of genome and structural ions in preventing viral capsid collapse during dehydration

Science.gov (United States)

Martín-González, Natalia; Guérin Darvas, Sofía M.; Durana, Aritz; Marti, Gerardo A.; Guérin, Diego M. A.; de Pablo, Pedro J.

2018-03-01

Even though viruses evolve mainly in liquid milieu, their horizontal transmission routes often include episodes of dry environment. Along their life cycle, some insect viruses, such as viruses from the Dicistroviridae family, withstand dehydrated conditions with presently unknown consequences to their structural stability. Here, we use atomic force microscopy to monitor the structural changes of viral particles of Triatoma virus (TrV) after desiccation. Our results demonstrate that TrV capsids preserve their genome inside, conserving their height after exposure to dehydrating conditions, which is in stark contrast with other viruses that expel their genome when desiccated. Moreover, empty capsids (without genome) resulted in collapsed particles after desiccation. We also explored the role of structural ions in the dehydration process of the virions (capsid containing genome) by chelating the accessible cations from the external solvent milieu. We observed that ion suppression helps to keep the virus height upon desiccation. Our results show that under drying conditions, the genome of TrV prevents the capsid from collapsing during dehydration, while the structural ions are responsible for promoting solvent exchange through the virion wall.

Assembly and characterization of foot-and-mouth disease virus empty capsid particles expressed within mammalian cells

DEFF Research Database (Denmark)

Gullberg, Maria; Muszynski, Bartosz; Organtini, Lindsey J.

2013-01-01

The foot-and-mouth disease virus (FMDV) structural protein precursor, P1-2A, is cleaved by the virus-encoded 3C protease (3Cpro) into the capsid proteins VP0, VP1 and VP3 (and 2A). In some systems, it is difficult to produce large amounts of these processed capsid proteins since 3Cpro can be toxic...... (from serotypes O and A) and 3Cpro were expressed from monocistronic cDNA cassettes as P1-2A-3C, or from dicistronic cassettes with the 3Cpro expression dependent on a mutant FMDV internal ribosome entry site (IRES) (designated P1-2A-mIRES-3C). The effects of using a mutant 3Cpro with reduced catalytic....... These products self-assembled to form FMDV empty capsid particles, which have a related, but distinct, morphology (as determined by electron microscopy and reconstruction) from that determined previously by X-ray crystallography. The assembled empty capsids bind, in a divalent cation-dependent manner, to the RGD...

Prevention and Control of Bed Bugs in Residences

Science.gov (United States)

... in other rooms, including bathrooms, living rooms and laundry rooms. Look for spots or smears Dark blood ... having the inspection done by a pest control service provider (exterminator). If you find a bed bug, ...

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

Relevance of Assembly-Activating Protein for Adeno-associated Virus Vector Production and Capsid Protein Stability in Mammalian and Insect Cells.

Science.gov (United States)

Grosse, Stefanie; Penaud-Budloo, Magalie; Herrmann, Anne-Kathrin; Börner, Kathleen; Fakhiri, Julia; Laketa, Vibor; Krämer, Chiara; Wiedtke, Ellen; Gunkel, Manuel; Ménard, Lucie; Ayuso, Eduard; Grimm, Dirk

2017-10-15

The discovery that adeno-associated virus 2 (AAV2) encodes an eighth protein, called assembly-activating protein (AAP), transformed our understanding of wild-type AAV biology. Concurrently, it raised questions about the role of AAP during production of recombinant vectors based on natural or molecularly engineered AAV capsids. Here, we show that AAP is indeed essential for generation of functional recombinant AAV2 vectors in both mammalian and insect cell-based vector production systems. Surprisingly, we observed that AAV2 capsid proteins VP1 to -3 are unstable in the absence of AAP2, likely due to rapid proteasomal degradation. Inhibition of the proteasome led to an increase of intracellular VP1 to -3 but neither triggered assembly of functional capsids nor promoted nuclear localization of the capsid proteins. Together, this underscores the crucial and unique role of AAP in the AAV life cycle, where it rapidly chaperones capsid assembly, thus preventing degradation of free capsid proteins. An expanded analysis comprising nine alternative AAV serotypes (1, 3 to 9, and rh10) showed that vector production always depends on the presence of AAP, with the exceptions of AAV4 and AAV5, which exhibited AAP-independent, albeit low-level, particle assembly. Interestingly, AAPs from all 10 serotypes could cross-complement AAP-depleted helper plasmids during vector production, despite there being distinct intracellular AAP localization patterns. These were most pronounced for AAP4 and AAP5, congruent with their inability to rescue an AAV2/AAP2 knockout. We conclude that AAP is key for assembly of genuine capsids from at least 10 different AAV serotypes, which has implications for vectors derived from wild-type or synthetic AAV capsids. IMPORTANCE Assembly of adeno-associated virus 2 (AAV2) is regulated by the assembly-activating protein (AAP), whose open reading frame overlaps with that of the viral capsid proteins. As the majority of evidence was obtained using virus

Characterization of Three Novel Linear Neutralizing B-Cell Epitopes in the Capsid Protein of Swine Hepatitis E Virus.

Science.gov (United States)

Chen, Yiyang; Liu, Baoyuan; Sun, Yani; Li, Huixia; Du, Taofeng; Nan, Yuchen; Hiscox, Julian A; Zhou, En-Min; Zhao, Qin

2018-04-18

Hepatitis E virus (HEV) causes liver disease in humans and is thought to be a zoonotic infection with domestic animals being a reservoir including swine and rabbits. One of the proteins encoded by the virus is the capsid protein. This is likely the major immune-dominant protein and a target for vaccination. Four monoclonal antibodies (MAbs); three novel; 1E4, 2C7, 2G9, and one previously characterized (1B5), were evaluated for binding to the capsid protein from genotype 4 (swine) hepatitis E virus (HEV). The results indicated that 625 DFCP 628 , 458 PSRPF 462 , and 407 EPTV 410 peptides on the capsid protein comprised minimal amino acid sequence motifs recognized by 1E4, 2C7, and 2G9, respectively. The data suggested that 2C7 and 2G9 epitopes were partially exposed on the surface of the capsid protein. Truncated genotype 4 swine HEV capsid protein (sp239, amino acids 368-606), can exist in multimeric forms. Pre-incubation of swine HEV with 2C7, 2G9, or 1B5 before addition to HepG2 cells partially blocked sp239 cell binding and inhibited swine HEV infection. The study indicated that 2C7, 2G9, and 1B5 partially blocked swine HEV infection of rabbits better than 1E4 or normal mouse IgG. The cross reactivity of antibodies suggested that capsid epitopes recognized by 2C7 and 2G9 are common to HEV strains infecting most host species. Collectively, MAbs 2C7, 2G9, and 1B5 were shown to recognize three novel linear neutralizing B-cell epitopes of genotype 4 HEV capsid protein. These results enhance understanding of HEV capsid protein structure to guide vaccine and anti-viral design. IMPORTANCE Genotype 3 and 4 HEVs are zoonotic viruses. Here, genotype 4 HEV was studied due to its prevalence in human populations and pig herds in China. To improve HEV disease diagnosis and prevention, a better understanding of antigenic structure and neutralizing epitopes of HEV capsid protein are needed. In this study, the locations of three novel linear B-cell recognition epitopes within

Bayesian Analysis for Penalized Spline Regression Using WinBUGS

Directory of Open Access Journals (Sweden)

Ciprian M. Crainiceanu

2005-09-01

Full Text Available Penalized splines can be viewed as BLUPs in a mixed model framework, which allows the use of mixed model software for smoothing. Thus, software originally developed for Bayesian analysis of mixed models can be used for penalized spline regression. Bayesian inference for nonparametric models enjoys the flexibility of nonparametric models and the exact inference provided by the Bayesian inferential machinery. This paper provides a simple, yet comprehensive, set of programs for the implementation of nonparametric Bayesian analysis in WinBUGS. Good mixing properties of the MCMC chains are obtained by using low-rank thin-plate splines, while simulation times per iteration are reduced employing WinBUGS specific computational tricks.

Cyclophilin A potentiates TRIM5α inhibition of HIV-1 nuclear import without promoting TRIM5α binding to the viral capsid.

Directory of Open Access Journals (Sweden)

Mallori Burse

Full Text Available The host immunophilin cyclophilin A (CypA binds to the capsid protein (CA of HIV-1 and regulates its infectivity. Depending on the target cell type, CypA can either promote or inhibit HIV-1 infection. The ability of CypA to promote HIV-1 infection has been extensively studied and linked to several steps in early replication including uncoating, reverse transcription and nuclear import. By contrast, the mechanism by which CypA inhibits infection is less well understood. We investigated the mechanism by which CypA potentiates restriction of HIV-1 by the tripartite motif-containing protein 5 (TRIM5α. Depletion of TRIM5α in the African green monkey cell line Vero, resulted in a loss of inhibition of infection by CypA, demonstrating that inhibition by CypA is mediated by TRIM5α. Complementary genetic and biochemical assays failed to demonstrate an ability of CypA to promote binding of TRIM5α to the viral capsid. TRIM5α inhibits HIV-1 reverse transcription in a proteasome-dependent manner; however, we observed that inhibition of proteasome activity did not reduce the ability of CypA to inhibit infection, suggesting that CypA acts at a step after reverse transcription. Accordingly, we observed a CypA-dependent reduction in the accumulation of nuclear HIV-1 DNA, indicating that CypA specifically promotes TRIM5α inhibition of HIV-1 nuclear import. We also observed that the ability of CypA to inhibit HIV-1 infection is abolished by amino acid substitutions within the conserved CPSF6-binding surface in CA. Our results indicate that CypA inhibits HIV-1 infection in Vero cells not by promoting TRIM5α binding to the capsid but by blocking nuclear import of the HIV-1 preintegration complex.

Unique features of a global human ectoparasite identified through sequencing of the bed bug genome.

Science.gov (United States)

Benoit, Joshua B; Adelman, Zach N; Reinhardt, Klaus; Dolan, Amanda; Poelchau, Monica; Jennings, Emily C; Szuter, Elise M; Hagan, Richard W; Gujar, Hemant; Shukla, Jayendra Nath; Zhu, Fang; Mohan, M; Nelson, David R; Rosendale, Andrew J; Derst, Christian; Resnik, Valentina; Wernig, Sebastian; Menegazzi, Pamela; Wegener, Christian; Peschel, Nicolai; Hendershot, Jacob M; Blenau, Wolfgang; Predel, Reinhard; Johnston, Paul R; Ioannidis, Panagiotis; Waterhouse, Robert M; Nauen, Ralf; Schorn, Corinna; Ott, Mark-Christoph; Maiwald, Frank; Johnston, J Spencer; Gondhalekar, Ameya D; Scharf, Michael E; Peterson, Brittany F; Raje, Kapil R; Hottel, Benjamin A; Armisén, David; Crumière, Antonin Jean Johan; Refki, Peter Nagui; Santos, Maria Emilia; Sghaier, Essia; Viala, Sèverine; Khila, Abderrahman; Ahn, Seung-Joon; Childers, Christopher; Lee, Chien-Yueh; Lin, Han; Hughes, Daniel S T; Duncan, Elizabeth J; Murali, Shwetha C; Qu, Jiaxin; Dugan, Shannon; Lee, Sandra L; Chao, Hsu; Dinh, Huyen; Han, Yi; Doddapaneni, Harshavardhan; Worley, Kim C; Muzny, Donna M; Wheeler, David; Panfilio, Kristen A; Vargas Jentzsch, Iris M; Vargo, Edward L; Booth, Warren; Friedrich, Markus; Weirauch, Matthew T; Anderson, Michelle A E; Jones, Jeffery W; Mittapalli, Omprakash; Zhao, Chaoyang; Zhou, Jing-Jiang; Evans, Jay D; Attardo, Geoffrey M; Robertson, Hugh M; Zdobnov, Evgeny M; Ribeiro, Jose M C; Gibbs, Richard A; Werren, John H; Palli, Subba R; Schal, Coby; Richards, Stephen

2016-02-02

The bed bug, Cimex lectularius, has re-established itself as a ubiquitous human ectoparasite throughout much of the world during the past two decades. This global resurgence is likely linked to increased international travel and commerce in addition to widespread insecticide resistance. Analyses of the C. lectularius sequenced genome (650 Mb) and 14,220 predicted protein-coding genes provide a comprehensive representation of genes that are linked to traumatic insemination, a reduced chemosensory repertoire of genes related to obligate hematophagy, host-symbiont interactions, and several mechanisms of insecticide resistance. In addition, we document the presence of multiple putative lateral gene transfer events. Genome sequencing and annotation establish a solid foundation for future research on mechanisms of insecticide resistance, human-bed bug and symbiont-bed bug associations, and unique features of bed bug biology that contribute to the unprecedented success of C. lectularius as a human ectoparasite.

Unique features of a global human ectoparasite identified through sequencing of the bed bug genome

Science.gov (United States)

Benoit, Joshua B.; Adelman, Zach N.; Reinhardt, Klaus; Dolan, Amanda; Poelchau, Monica; Jennings, Emily C.; Szuter, Elise M.; Hagan, Richard W.; Gujar, Hemant; Shukla, Jayendra Nath; Zhu, Fang; Mohan, M.; Nelson, David R.; Rosendale, Andrew J.; Derst, Christian; Resnik, Valentina; Wernig, Sebastian; Menegazzi, Pamela; Wegener, Christian; Peschel, Nicolai; Hendershot, Jacob M.; Blenau, Wolfgang; Predel, Reinhard; Johnston, Paul R.; Ioannidis, Panagiotis; Waterhouse, Robert M.; Nauen, Ralf; Schorn, Corinna; Ott, Mark-Christoph; Maiwald, Frank; Johnston, J. Spencer; Gondhalekar, Ameya D.; Scharf, Michael E.; Peterson, Brittany F.; Raje, Kapil R.; Hottel, Benjamin A.; Armisén, David; Crumière, Antonin Jean Johan; Refki, Peter Nagui; Santos, Maria Emilia; Sghaier, Essia; Viala, Sèverine; Khila, Abderrahman; Ahn, Seung-Joon; Childers, Christopher; Lee, Chien-Yueh; Lin, Han; Hughes, Daniel S. T.; Duncan, Elizabeth J.; Murali, Shwetha C.; Qu, Jiaxin; Dugan, Shannon; Lee, Sandra L.; Chao, Hsu; Dinh, Huyen; Han, Yi; Doddapaneni, Harshavardhan; Worley, Kim C.; Muzny, Donna M.; Wheeler, David; Panfilio, Kristen A.; Vargas Jentzsch, Iris M.; Vargo, Edward L.; Booth, Warren; Friedrich, Markus; Weirauch, Matthew T.; Anderson, Michelle A. E.; Jones, Jeffery W.; Mittapalli, Omprakash; Zhao, Chaoyang; Zhou, Jing-Jiang; Evans, Jay D.; Attardo, Geoffrey M.; Robertson, Hugh M.; Zdobnov, Evgeny M.; Ribeiro, Jose M. C.; Gibbs, Richard A.; Werren, John H.; Palli, Subba R.; Schal, Coby; Richards, Stephen

2016-01-01

The bed bug, Cimex lectularius, has re-established itself as a ubiquitous human ectoparasite throughout much of the world during the past two decades. This global resurgence is likely linked to increased international travel and commerce in addition to widespread insecticide resistance. Analyses of the C. lectularius sequenced genome (650 Mb) and 14,220 predicted protein-coding genes provide a comprehensive representation of genes that are linked to traumatic insemination, a reduced chemosensory repertoire of genes related to obligate hematophagy, host–symbiont interactions, and several mechanisms of insecticide resistance. In addition, we document the presence of multiple putative lateral gene transfer events. Genome sequencing and annotation establish a solid foundation for future research on mechanisms of insecticide resistance, human–bed bug and symbiont–bed bug associations, and unique features of bed bug biology that contribute to the unprecedented success of C. lectularius as a human ectoparasite. PMID:26836814

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.

Effects of immunosuppression on circulating adeno-associated virus capsid-specific T cells in humans.

Science.gov (United States)

Parzych, Elizabeth M; Li, Hua; Yin, Xiangfan; Liu, Qin; Wu, Te-Lang; Podsakoff, Gregory M; High, Katherine A; Levine, Matthew H; Ertl, Hildegund C J

2013-04-01

In humans adeno-associated virus (AAV)-mediated gene transfer is followed by expansion of AAV capsid-specific T cells, evidence of cell damage, and loss of transgene product expression, implicating immunological rejection of vector-transduced cells, which may be prevented by immunosuppressive drugs. We undertook this study to assess the effect of immunosuppression (IS) used for organ transplantation on immune responses to AAV capsid antigens. Recipients of liver or kidney transplants were tested before and 4 weeks after induction of IS in comparison with matched samples from healthy human adults and an additional cohort with comorbid conditions similar to those of the transplant patients. Our data show that transplant patients and comorbid control subjects have markedly higher frequencies of circulating AAV capsid-specific T cells compared with healthy adults. On average, IS resulted in a reduction of AAV-specific CD4⁺ T cells, whereas numbers of circulating CD8⁺ effector and central memory T cells tended to increase. Independent of the type of transplant or the IS regimens, the trend of AAV capsid-specific T cell responses after drug treatment varied; in some patients responses were unaffected whereas others showed decreases or even pronounced increases, casting doubt on the usefulness of prophylactic IS for AAV vector recipients.

MENGGALI METRIK YANG PENTING DALAM RANGKAIAN AKTIVITAS PERUBAHAN PERANGKAT LUNAK UNTUK MEMPREDIKSI BUG DENGAN ATURAN ASOSIASI

Directory of Open Access Journals (Sweden)

Achmad Arwan

2014-01-01

Full Text Available Normal 0 false false false IN X-NONE X-NONE MicrosoftInternetExplorer4 /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0cm 5.4pt 0cm 5.4pt; mso-para-margin:0cm; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:"Times New Roman"; mso-bidi-theme-font:minor-bidi;} Dalam evolusi perangkat lunak terdapat rangkaian aktivitas proses perubahan yang kemudian oleh Nachiapan diformulasikan menjadi metrik-metrik yang mampu memprediksi bug secara presisi. Akan tetapi secara spesifik belum ditemukan seberapa besar pengaruh masing-masing metrik tersebut terhadap hasil prediksi sebuah bug. Penelitian ini mengusulkan pengukuran jumlah kemunculan metrik dengan kemunculan bug pada proyek eclipse. Sehingga dengan penelitian ini dapat diketahui metrik -metrik mana yang penting dalam prediksi kemunculan bug. Aturan asosiasi dalam penggalian data telah dipergunakan secara luas untuk menggali variabel-variabel saling terkait dalam sampel data. Metrik-metrik dalam proyek eclipse kemudian digali dengan aturan asosiasi untuk mendapatkan metrik yang muncul bersama bug. Hasil pengujian menunjukkan bahwa metrik Numberofchangeslate rata-rata kemunculannya sebesar 46,9%, sedangkan metrik Peopletotal muncul bersama bug rata-rata sebesar 43,57% dan metrik Numberofchangesearly memiliki kemunculan rata-rata 14% sehingga ketiga metrik tersebut merupakan metrik yang penting dalam memprediksi bug.

Hiring a Pest Management Professional for Bed Bugs

Science.gov (United States)

If you hire someone to treat your bed bug infestation, make sure they use Integrated Pest Management (IPM) techniques, check credentials, and know they may need multiple visits, to take apart furniture, and to use vacuums, heat, and pesticides.

Four levels of hierarchical organization, including noncovalent chainmail, brace the mature tumor herpesvirus capsid against pressurization.

Science.gov (United States)

Zhou, Z Hong; Hui, Wong Hoi; Shah, Sanket; Jih, Jonathan; O'Connor, Christine M; Sherman, Michael B; Kedes, Dean H; Schein, Stan

2014-10-07

Like many double-stranded DNA viruses, tumor gammaherpesviruses Epstein-Barr virus and Kaposi's sarcoma-associated herpesvirus withstand high internal pressure. Bacteriophage HK97 uses covalent chainmail for this purpose, but how this is achieved noncovalently in the much larger gammaherpesvirus capsid is unknown. Our cryoelectron microscopy structure of a gammaherpesvirus capsid reveals a hierarchy of four levels of organization: (1) Within a hexon capsomer, each monomer of the major capsid protein (MCP), 1,378 amino acids and six domains, interacts with its neighboring MCPs at four sites. (2) Neighboring capsomers are linked in pairs by MCP dimerization domains and in groups of three by heterotrimeric triplex proteins. (3) Small (∼280 amino acids) HK97-like domains in MCP monomers alternate with triplex heterotrimers to form a belt that encircles each capsomer. (4) One hundred sixty-two belts concatenate to form noncovalent chainmail. The triplex heterotrimer orchestrates all four levels and likely drives maturation to an angular capsid that can withstand pressurization. Copyright © 2014 Elsevier Ltd. All rights reserved.

Edge effects of stink bugs in corn, cotton, peanut and soybean fields adjacent to woodland.

Science.gov (United States)

Producers face significant crop losses from stink bug species in the southeastern USA, but the high mobility and polyphagy of the bugs make predictions of their presence in crops difficult. While there is some evidence that they colonize crops from adjacent crops, there are no studies of their colo...

Economic Injury Level of the Neotropical Brown Stink Bug Euschistus heros (F.) on Cotton Plants.

Science.gov (United States)

Soria, M F; Degrande, P E; Panizzi, A R; Toews, M D

2017-06-01

In Brazil, the Neotropical brown stink bug, Euschistus heros (F.) (Hemiptera: Pentatomidae), commonly disperses from soybeans to cotton fields. The establishment of an economic treatment threshold for this pest on cotton crops is required. Infestation levels of adults of E. heros were evaluated on cotton plants at preflowering, early flowering, boll filling, and full maturity by assessing external and internal symptoms of injury on bolls, seed cotton/lint production, and fiber quality parameters. A completely randomized experiment was designed to infest cotton plants in a greenhouse with 0, 2, 4, 6, and 8 bugs/plant, except at the full-maturity stage in which only infestation with 8 bugs/plant and uninfested plants were evaluated. Results indicated that the preflowering, early-flowering, and full-maturity stages were not affected by E. heros. A linear regression model showed a significant increase in the number of internal punctures and warts in the boll-filling stage as the population of bugs increased. The average number of loci with mottled immature fibers was significantly higher at 4, 6, and 8 bugs compared with uninfested plants with data following a quadratic regression model. The seed and lint cotton was reduced by 18 and 25% at the maximum level of infestation (ca. 8 bugs/plant) in the boll-filling stage. The micronaire and yellowing indexes were, respectively, reduced and increased with the increase of the infestation levels. The economic injury level of E. heros on cotton plants at the boll-filling stage was determined as 0.5 adult/plant. Based on that, a treatment threshold of 0.1 adult/plant can be recommended to avoid economic losses.

Plus- and minus-end directed microtubule motors bind simultaneously to herpes simplex virus capsids using different inner tegument structures.

Directory of Open Access Journals (Sweden)

Kerstin Radtke

2010-07-01

Full Text Available Many viruses depend on host microtubule motors to reach their destined intracellular location. Viral particles of neurotropic alphaherpesviruses such as herpes simplex virus 1 (HSV1 show bidirectional transport towards the cell center as well as the periphery, indicating that they utilize microtubule motors of opposing directionality. To understand the mechanisms of specific motor recruitment, it is necessary to characterize the molecular composition of such motile viral structures. We have generated HSV1 capsids with different surface features without impairing their overall architecture, and show that in a mammalian cell-free system the microtubule motors dynein and kinesin-1 and the dynein cofactor dynactin could interact directly with capsids independent of other host factors. The capsid composition and surface was analyzed with respect to 23 structural proteins that are potentially exposed to the cytosol during virus assembly or cell entry. Many of these proteins belong to the tegument, the hallmark of all herpesviruses located between the capsid and the viral envelope. Using immunoblots, quantitative mass spectrometry and quantitative immunoelectron microscopy, we show that capsids exposing inner tegument proteins such as pUS3, pUL36, pUL37, ICP0, pUL14, pUL16, and pUL21 recruited dynein, dynactin, kinesin-1 and kinesin-2. In contrast, neither untegumented capsids exposing VP5, VP26, pUL17 and pUL25 nor capsids covered by outer tegument proteins such as vhs, pUL11, ICP4, ICP34.5, VP11/12, VP13/14, VP16, VP22 or pUS11 bound microtubule motors. Our data suggest that HSV1 uses different structural features of the inner tegument to recruit dynein or kinesin-1. Individual capsids simultaneously accommodated motors of opposing directionality as well as several copies of the same motor. Thus, these associated motors either engage in a tug-of-war or their activities are coordinately regulated to achieve net transport either to the nucleus during

Protecting Yourself from Bed Bugs in Public Places

Science.gov (United States)

... office. By reducing clutter in your workplace or school, you provide fewer places for the bed bugs to hide and fewer opportunities for them to hitchhike to your home. Keep your ... office or school, consider storing your belongings in a plastic bin. ...

Perspective on Adeno-Associated Virus Capsid Modification for Duchenne Muscular Dystrophy Gene Therapy.

Science.gov (United States)

Nance, Michael E; Duan, Dongsheng

2015-12-01

Duchenne muscular dystrophy (DMD) is a X-linked, progressive childhood myopathy caused by mutations in the dystrophin gene, one of the largest genes in the genome. It is characterized by skeletal and cardiac muscle degeneration and dysfunction leading to cardiac and/or respiratory failure. Adeno-associated virus (AAV) is a highly promising gene therapy vector. AAV gene therapy has resulted in unprecedented clinical success for treating several inherited diseases. However, AAV gene therapy for DMD remains a significant challenge. Hurdles for AAV-mediated DMD gene therapy include the difficulty to package the full-length dystrophin coding sequence in an AAV vector, the necessity for whole-body gene delivery, the immune response to dystrophin and AAV capsid, and the species-specific barriers to translate from animal models to human patients. Capsid engineering aims at improving viral vector properties by rational design and/or forced evolution. In this review, we discuss how to use the state-of-the-art AAV capsid engineering technologies to overcome hurdles in AAV-based DMD gene therapy.

AAV capsid CD8+ T-cell epitopes are highly conserved across AAV serotypes.

Science.gov (United States)

Hui, Daniel J; Edmonson, Shyrie C; Podsakoff, Gregory M; Pien, Gary C; Ivanciu, Lacramioara; Camire, Rodney M; Ertl, Hildegund; Mingozzi, Federico; High, Katherine A; Basner-Tschakarjan, Etiena

2015-01-01

Adeno-associated virus (AAV) has become one of the most promising vectors in gene transfer in the last 10 years with successful translation to clinical trials in humans and even market approval for a first gene therapy product in Europe. Administration to humans, however, revealed that adaptive immune responses against the vector capsid can present an obstacle to sustained transgene expression due to the activation and expansion of capsid-specific T cells. The limited number of peripheral blood mononuclear cells (PBMCs) obtained from samples within clinical trials allows for little more than monitoring of T-cell responses. We were able to identify immunodominant major histocompatibility complex (MHC) class I epitopes for common human leukocyte antigen (HLA) types by using spleens isolated from subjects undergoing splenectomy for non-malignant indications as a source of large numbers of lymphocytes and restimulating them with single AAV capsid peptides in vitro. Further experiments confirmed that these epitopes are naturally processed and functionally relevant. The design of more effective and less immunogenic AAV vectors, and precise immune monitoring of vector-infused subjects, are facilitated by these findings.

Series of Norovirus Outbreaks Caused by Consumption of Green Coral Lettuce, Denmark, April 2016

DEFF Research Database (Denmark)

Müller, Luise; Rasmussen, Lasse Dam; Jensen, Tenna

2016-01-01

In early April 2016, an unusual high number of point-source outbreaks of gastrointestinal disease were reported to occur in Denmark. Outbreaks were individually investigated. Two analytical studies were performed. Patient stool samples collected and analysed; positive stool samples were sequenced...... over the polymerase and/or capsid gene areas. Implicated lettuce heads were collected and analysed for the presence of norovirus. Foods were traced-back and traced-forward and international alert systems applied. A total of 23 linked point-source outbreaks occurred over the course of one week. Fresh...... green coral lettuce (Lollo Bionda lettuce) had been consumed in all settings. In a cohort study including 234 participants a dish containing green lettuce was associated with illness. Norovirus of Genogroup I (GI) was detected in samples from 28 patients comprising eight of the outbreaks. Sequencing...

Verde plant bug, Creontiades signatus (Hemiptera: Miridae) effects of insect density and bloom period of infestation on cotton damage and yield

Science.gov (United States)

The verde plant bug, Creontiades signatus Distant (Hemiptera: Miridae), has emerged as a threat to cotton in South Texas, causing boll damage similar to boll-feeding stink bugs (Hemiptera: Pentatomidae). Verde plant bugs were released into caged cotton for a one-week period to characterize the effec...

In vitro protease cleavage and computer simulations reveal the HIV-1 capsid maturation pathway

Science.gov (United States)

Ning, Jiying; Erdemci-Tandogan, Gonca; Yufenyuy, Ernest L.; Wagner, Jef; Himes, Benjamin A.; Zhao, Gongpu; Aiken, Christopher; Zandi, Roya; Zhang, Peijun

2016-12-01

HIV-1 virions assemble as immature particles containing Gag polyproteins that are processed by the viral protease into individual components, resulting in the formation of mature infectious particles. There are two competing models for the process of forming the mature HIV-1 core: the disassembly and de novo reassembly model and the non-diffusional displacive model. To study the maturation pathway, we simulate HIV-1 maturation in vitro by digesting immature particles and assembled virus-like particles with recombinant HIV-1 protease and monitor the process with biochemical assays and cryoEM structural analysis in parallel. Processing of Gag in vitro is accurate and efficient and results in both soluble capsid protein and conical or tubular capsid assemblies, seemingly converted from immature Gag particles. Computer simulations further reveal probable assembly pathways of HIV-1 capsid formation. Combining the experimental data and computer simulations, our results suggest a sequential combination of both displacive and disassembly/reassembly processes for HIV-1 maturation.

BIOLOGICAL FEATURES OF TARAN (RUTILUS HECKELII OF THE DNIEPER-BUG ESTUARY SYSTEM

Directory of Open Access Journals (Sweden)

K. Geina

2016-03-01

Full Text Available Purpose. To analyze morphological variability and basic biological features of the modern stock of taran of the Dnieper-Bug estuary system in the conditions of the transformed Dnieper flow. Methodology. Morphological variability was determined based on the comparison of morphological features of roach Rutilus rutilus (Linnaeus, 1758 of the Kakhovka reservoir and semi-migratory taran Rutilus rutilus heckelii (Nordmann, 1840 of the Dnieper-Bug estuary system. As a main criterion of the evaluation of the taran stock biological state, we used age, sex structure, growth characteristics, fecundity and condition factor. Fish sampling was carried out at stationary monitoring-observation stations of the Institute of Fisheries NAAS of Ukraine. Field and cameral processing of the collected materials were performed based on conventional methods and guidelines. Findings. Morphological variability analysis demonstrated significant differences between Dnieper and Bug taran. The highest differences were observed for antroventral distance (td=11.19. Significant differences were also observed for antenanal and ventroanal distances td=4.05-4.14. No significant differences were found for meristic features. There were also significant differences between Dnieper-Bug taran and Kakhovka reservoir roach, which had formed a resident form after regulating the Dnieper River flow. Kakhovka reservoir roach is more deep-bodied with Н=32.79%, t-test value was 5.65. Pelvic fins were more shifted to the caudal fin (td=5.28 that resulted in significant difference (p<0.05 in ventroanal distance (td=4.26. Taran also had somewhat smaller length of the anal fin base (td=4.73 but its height was higher – td=5.78. The main peculiarity of the current biological state of taran stock is the domination of young age groups. The small number of fish in the boundary groups of the age series right wing with relative stability of growth features indicate on intensive pressure on the

Transient Bluetongue virus serotype 8 capsid protein expression in Nicotiana benthamiana

Directory of Open Access Journals (Sweden)

Albertha R. van Zyl

2016-03-01

Full Text Available Bluetongue virus (BTV causes severe disease in domestic and wild ruminants, and has recently caused several outbreaks in Europe. Current vaccines include live-attenuated and inactivated viruses; while these are effective, there is risk of reversion to virulence by mutation or reassortment with wild type viruses. Subunit or virus-like particle (VLP vaccines are safer options: VLP vaccines produced in insect cells by expression of the four BTV capsid proteins are protective against challenge; however, this is a costly production method. We investigated production of BTV VLPs in plants via Agrobacterium-mediated transient expression, an inexpensive production system very well suited to developing country use. Leaves infiltrated with recombinant pEAQ-HT vectors separately encoding the four BTV-8 capsid proteins produced more proteins than recombinant pTRA vectors. Plant expression using the pEAQ-HT vector resulted in both BTV-8 core-like particles (CLPs and VLPs; differentially controlling the concentration of infiltrated bacteria significantly influenced yield of the VLPs. In situ localisation of assembled particles was investigated by using transmission electron microscopy (TEM and it was shown that a mixed population of core-like particles (CLPs, consisting of VP3 and VP7 and VLPs were present as paracrystalline arrays in the cytoplasm of plant cells co-expressing all four capsid proteins.

Viral capsid is a pathogen-associated molecular pattern in adenovirus keratitis.

Directory of Open Access Journals (Sweden)

Ashish V Chintakuntlawar

2010-04-01

Full Text Available Human adenovirus (HAdV infection of the human eye, in particular serotypes 8, 19 and 37, induces the formation of corneal subepithelial leukocytic infiltrates. Using a unique mouse model of adenovirus keratitis, we studied the role of various virus-associated molecular patterns in subsequent innate immune responses of resident corneal cells to HAdV-37 infection. We found that neither viral DNA, viral gene expression, or viral replication was necessary for the development of keratitis. In contrast, empty viral capsid induced keratitis and a chemokine profile similar to intact virus. Transfected viral DNA did not induce leukocyte infiltration despite CCL2 expression similar to levels in virus infected corneas. Mice without toll-like receptor 9 (Tlr9 signaling developed clinical keratitis upon HAdV-37 infection similar to wild type mice, although the absolute numbers of activated monocytes in the cornea were less in Tlr9(-/- mice. Virus induced leukocytic infiltrates and chemokine expression in mouse cornea could be blocked by treatment with a peptide containing arginine glycine aspartic acid (RGD. These results demonstrate that adenovirus infection of the cornea induces chemokine expression and subsequent infiltration by leukocytes principally through RGD contact between viral capsid and the host cell, possibly through direct interaction between the viral capsid penton base and host cell integrins.

Molecular characterization of capsid protein gene of potato virus X ...

African Journals Online (AJOL)

Molecular characterization of capsid protein gene of potato virus X from Pakistan. Arshad Jamal, Idrees Ahmad Nasir, Bushra Tabassum, Muhammad Tariq, Abdul Munim Farooq, Zahida Qamar, Mohsin Ahmad Khan, Nadeem Ahmad, Muhammad Shafiq, Muhammad Saleem Haider, M. Arshad Javed, Tayyab Husnain ...

Bug bites and stings: When to see a dermatologist

Medline Plus

Full Text Available ... Part 1: Structure Part 2: Origin Part 3: Function Textbook Study notes Image library 3-D animated ... nail care Injured skin Bug bites and stings "); (function () { var a = "", b = [ "adid=aad-aad-1", "site= ...

Bug bites and stings: When to see a dermatologist

Medline Plus

Full Text Available ... aging skin care Kids’ zone About skin: Your body's largest organ About hair: Not just on your ... bug bite, such as a rash, fever, or body aches, see your doctor or a board-certified ...

Bug bites and stings: When to see a dermatologist

Medline Plus

Full Text Available ... bites Although most bug bites are harmless, some can spread dangerous diseases like Zika virus, dengue, Lyme ... and tuck your shirt into your pants. You can also pre-treat outer layers of clothing with ...

Bug bites and stings: When to see a dermatologist

Medline Plus

Full Text Available ... and treatments Acne and rosacea Bumps and growths Color problems Contagious skin diseases Cosmetic treatments Dry / sweaty ... prevent bug bites, dermatologists recommend the following tips: Use insect repellent. To protect against mosquitoes, ticks and ...

Bug bites and stings: When to see a dermatologist

Medline Plus

Full Text Available ... a dermatologist Why see a board-certified dermatologist? Home Public and patients Skin, hair, and nail care ... bites and stings can be safely treated at home. To treat bug bites and stings at home, ...

Intravenous administration of the adeno-associated virus-PHP.B capsid fails to upregulate transduction efficiency in the marmoset brain.

Science.gov (United States)

Matsuzaki, Yasunori; Konno, Ayumu; Mochizuki, Ryuta; Shinohara, Yoichiro; Nitta, Keisuke; Okada, Yukihiro; Hirai, Hirokazu

2018-02-05

Intravenous administration of adeno-associated virus (AAV)-PHP.B, a capsid variant of AAV9 containing seven amino acid insertions, results in a greater permeability of the blood brain barrier (BBB) than standard AAV9 in mice, leading to highly efficient and global transduction of the central nervous system (CNS). The present study aimed to examine whether the enhanced BBB penetrance of AAV-PHP.B observed in mice also occurs in non-human primates. Thus, a young adult (age, 1.6 years) and an old adult (age, 7.2 years) marmoset received an intravenous injection of AAV-PHP.B expressing enhanced green fluorescent protein (EGFP) under the control of the constitutive CBh promoter (a hybrid of cytomegalovirus early enhancer and chicken β-actin promoter). Age-matched control marmosets were treated with standard AAV9-capsid vectors. The animals were sacrificed 6 weeks after the viral injection. Based on the results, only limited transduction of neurons (0-2%) and astrocytes (0.1-2.5%) was observed in both AAV-PHP.B- and AAV9-treated marmosets. One noticeable difference between AAV-PHP.B and AAV9 was the marked transduction of the peripheral dorsal root ganglia neurons. Indeed, the soma and axons in the projection from the spinal cord to the nucleus cuneatus in the medulla oblongata were strongly labeled with EGFP by AAV-PHP.B. Thus, except for the peripheral dorsal root ganglia neurons, the AAV-PHP.B transduction efficiency in the CNS of marmosets was comparable to that of AAV9 vectors. Copyright © 2017 Elsevier B.V. All rights reserved.

Biology, Pest Status, Microbiome and Control of Kudzu Bug (Hemiptera: Heteroptera: Plataspidae): A New Invasive Pest in the U.S.

Science.gov (United States)

Dhammi, Anirudh; van Krestchmar, Jaap B.; Ponnusamy, Loganathan; Bacheler, Jack S.; Reisig, Dominic D.; Herbert, Ames; Del Pozo-Valdivia, Alejandro I.; Roe, R. Michael

2016-01-01

Soybean is an important food crop, and insect integrated pest management (IPM) is critical to the sustainability of this production system. In recent years, the introduction into the United States of the kudzu bug currently identified as Megacopta cribraria (F.), poses a threat to soybean production. The kudzu bug was first discovered in the state of Georgia, U.S. in 2009 and since then has spread to most of the southeastern states. Because it was not found in the North American subcontinent before this time, much of our knowledge of this insect comes from research done in its native habitat. However, since the U.S. introduction, studies have been undertaken to improve our understanding of the kudzu bug basic biology, microbiome, migration patterns, host selection and management in its expanding new range. Researchers are not only looking at developing IPM strategies for the kudzu bug in soybean, but also at its unique relationship with symbiotic bacteria. Adult females deposit bacterial packets with their eggs, and the neonates feed on these packets to acquire the bacteria, Candidatus Ishikawaella capsulata. The kudzu bug should be an informative model to study the co-evolution of insect function and behavior with that of a single bacteria species. We review kudzu bug trapping and survey methods, the development of bioassays for insecticide susceptibility, insecticide efficacy, host preferences, impact of the pest on urban environments, population expansion, and the occurrence of natural enemies. The identity of the kudzu bug in the U.S. is not clear. We propose that the kudzu bug currently accepted as M. cribraria in the U.S. is actually Megacopta punctatissima, with more work needed to confirm this hypothesis. PMID:27649166

Bug bites and stings: When to see a dermatologist

Medline Plus

Full Text Available ... Particularly if you’re visiting areas with known insect-borne diseases, it’s important to take steps to ... bug bites, dermatologists recommend the following tips: Use insect repellent. To protect against mosquitoes, ticks and other ...

Bug bites and stings: When to see a dermatologist

Medline Plus

Full Text Available ... Choosing Wisely Education Online Learning Center MOC Recognized Credit Basic Derm Curriculum Teaching and learning guides Suggested ... it’s important to take steps to reduce your risk. To help prevent bug bites, dermatologists recommend the ...

Bug bites and stings: When to see a dermatologist

Medline Plus

Full Text Available ... and Lectureship Clarence S. Livingood Award and Lectureship Marion B. Sulzberger Award and Lectureship Master Dermatologist Award Members ... skin Bug bites and stings "); (function () { var a = "", b = [ "adid=aad-aad-1", "site=ehs.con.aad. ...

Random Insertion of mCherry Into VP3 Domain of Adeno-associated Virus Yields Fluorescent Capsids With no Loss of Infectivity

Directory of Open Access Journals (Sweden)

Justin Judd

2012-01-01

Full Text Available Adeno-associated virus (AAV-derived vectors are promising gene delivery systems, and a number of design strategies have been pursued to improve their performance. For example, genetic insertion of proteins into the capsid may be used to achieve vector retargeting, reduced immunogenicity, or to track vector transport. Unfortunately, rational approaches to genetic insertion have experienced limited success due to the unpredictable context-dependent nature of protein folding and the complexity of the capsid's macroassembly. We report the construction and use of a frame-enriched DNase-based random insertion library based on AAV2 cap, called pAAV2_RaPID (Random Peptide Insertion by DNase. The fluorescent mCherry protein was inserted randomly throughout the AAV2 capsid and the library was selected for fluorescent and infectious variants. A capsid site was identified in VP3 that can tolerate the large protein insertion. In contrast to previous efforts to incorporate fluorescent proteins into the AAV2 capsid, the isolated mCherry mutant maintains native infectivity while displaying robust fluorescence. Collectively, these results demonstrate that the pAAV2_RaPID platform library can be used to create fully infectious AAV vectors carrying large functional protein domains on the capsid.

Recolonization of bat roost by bat bugs (Cimex pipistrelli): could parasite load be a cause of bat roost switching?

Science.gov (United States)

Bartonička, Tomáš; Růžičková, Lucie

2013-04-01

Roost ectoparasites are believed to have a negative impact on fitness of their hosts as birds or mammals. Previous studies were mostly focussed on the synchronization between reproduction cycles of ectoparasites and hosts living in infested roosts. However, to date, it has not been examined how fast ectoparasites colonize new, non-infested roosts and thus increasing the impact on the local populations of hosts. The parasite-host model was studied, including bat bugs Cimex pipistrelli and soprano pipistrelles Pipistrellus pygmaeus, where bat behaviour was observed which tended to reduce the parasite load in bat roosts. We investigated (1) whether bats change their roosting behaviour when we discontinued synchronization of their reproduction and the life cycle of the bat bugs and (2) how fast and which stages of bat bugs reoccupy cleaned roosts. In a 3-year field experiment, we removed all bat bugs from six bat boxes in each spring. Pipistrelles bred young in all non-infested boxes during these 3 years. In addition, 8 years of regular observations before this experiment indicate that bats avoided breeding in the same bat boxes at all. Bat bugs were found again in clean boxes in mid-May. However, their densities did not maximise before the beginning of June, before parturition. A re-appearance of bugs was observed after 21-56 days after the first bat visit. Adult bugs, mainly females, colonised cleaned boxes first though at the same time there were a lot of younger and smaller instars in non-manipulated roosts in the vicinity.

Big-Eyed Bugs Have Big Appetite for Pests

Science.gov (United States)

Many kinds of arthropod natural enemies (predators and parasitoids) inhabit crop fields in Arizona and can have a large negative impact on several pest insect species that also infest these crops. Geocoris spp., commonly known as big-eyed bugs, are among the most abundant insect predators in field c...

Bug bites and stings: When to see a dermatologist

Medline Plus

Full Text Available ... hair, and nail care Skin care Hair care / hair loss Injured skin Nail care Anti-aging skin care ... hair, and nail care Skin care Hair care / hair loss Injured skin Blisters Bug bites and stings How ...

Bug bites and stings: When to see a dermatologist

Medline Plus

Full Text Available ... care Injured skin Bug bites and stings "); (function () { var a = "", b = [ "adid=aad-aad-1", "site=ehs. ... t", "hpos=l", "zone=public", "mcat1=de12", ]; for (var c = 0; c "); (function () { var a = "", b = [ "adid= ...

Imaging and Quantitation of a Succession of Transient Intermediates Reveal the Reversible Self-Assembly Pathway of a Simple Icosahedral Virus Capsid.

Science.gov (United States)

Medrano, María; Fuertes, Miguel Ángel; Valbuena, Alejandro; Carrillo, Pablo J P; Rodríguez-Huete, Alicia; Mateu, Mauricio G

2016-11-30

Understanding the fundamental principles underlying supramolecular self-assembly may facilitate many developments, from novel antivirals to self-organized nanodevices. Icosahedral virus particles constitute paradigms to study self-assembly using a combination of theory and experiment. Unfortunately, assembly pathways of the structurally simplest virus capsids, those more accessible to detailed theoretical studies, have been difficult to study experimentally. We have enabled the in vitro self-assembly under close to physiological conditions of one of the simplest virus particles known, the minute virus of mice (MVM) capsid, and experimentally analyzed its pathways of assembly and disassembly. A combination of electron microscopy and high-resolution atomic force microscopy was used to structurally characterize and quantify a succession of transient assembly and disassembly intermediates. The results provided an experiment-based model for the reversible self-assembly pathway of a most simple (T = 1) icosahedral protein shell. During assembly, trimeric capsid building blocks are sequentially added to the growing capsid, with pentamers of building blocks and incomplete capsids missing one building block as conspicuous intermediates. This study provided experimental verification of many features of self-assembly of a simple T = 1 capsid predicted by molecular dynamics simulations. It also demonstrated atomic force microscopy imaging and automated analysis, in combination with electron microscopy, as a powerful single-particle approach to characterize at high resolution and quantify transient intermediates during supramolecular self-assembly/disassembly reactions. Finally, the efficient in vitro self-assembly achieved for the oncotropic, cell nucleus-targeted MVM capsid may facilitate its development as a drug-encapsidating nanoparticle for anticancer targeted drug delivery.

A Transcriptome Survey Spanning Life Stages and Sexes of the Harlequin Bug, Murgantia histrionica

Directory of Open Access Journals (Sweden)

Michael E. Sparks

2017-05-01

Full Text Available The harlequin bug, Murgantia histrionica (Hahn, is an agricultural pest in the continental United States, particularly in southern states. Reliable gene sequence data are especially useful to the development of species-specific, environmentally friendly molecular biopesticides and effective biolures for this insect. Here, mRNAs were sampled from whole insects at the 2nd and 4th nymphal instars, as well as sexed adults, and sequenced using Illumina RNA-Seq technology. A global assembly of these data identified 72,540 putative unique transcripts bearing high levels of similarity to transcripts identified in other taxa, with over 99% of conserved single-copy orthologs among insects being detected. Gene ontology and protein family analyses were conducted to explore the functional potential of the harlequin bug’s gene repertoire, and phylogenetic analyses were conducted on gene families germane to xenobiotic detoxification, including glutathione S-transferases, carboxylesterases and cytochrome P450s. Genic content in harlequin bug was compared with that of the closely related invasive pest, the brown marmorated stink bug, Halyomorpha halys (Stål. Quantitative analyses of harlequin bug gene expression levels, experimentally validated using quantitative real-time PCR, identified genes differentially expressed between life stages and/or sexes.

Pheromone of the banana-spotting bug, amblypelta lutescens lutescens Distant (Hemiptera: Coreidae): identification, synthesis and field bioassay

Science.gov (United States)

The banana spotting bug Amblypelta lutescens lutescens Distant (Hemiptera: Coreidae) is one of the principal pests of tree fruits and nuts across northern and eastern Australia. Apart from damage assessments in orchards, there are currently no other methods for monitoring bug activity to aid manage...

2016 All Bugs Good and Bad Webinar Series - eXtension

Science.gov (United States)

Importadas Resources Instructional Videos Publications Top of the Mound Game Fire Ant Images from BugMugs.org Breeding and Genomics Sheep Small and Backyard Flocks Small Meat Processors Sustainable Marine Fisheries

AAV capsid CD8+ T-cell epitopes are highly conserved across AAV serotypes

Directory of Open Access Journals (Sweden)

Daniel J Hui

Full Text Available Adeno-associated virus (AAV has become one of the most promising vectors in gene transfer in the last 10 years with successful translation to clinical trials in humans and even market approval for a first gene therapy product in Europe. Administration to humans, however, revealed that adaptive immune responses against the vector capsid can present an obstacle to sustained transgene expression due to the activation and expansion of capsid-specific T cells. The limited number of peripheral blood mononuclear cells (PBMCs obtained from samples within clinical trials allows for little more than monitoring of T-cell responses. We were able to identify immunodominant major histocompatibility complex (MHC class I epitopes for common human leukocyte antigen (HLA types by using spleens isolated from subjects undergoing splenectomy for non-malignant indications as a source of large numbers of lymphocytes and restimulating them with single AAV capsid peptides in vitro. Further experiments confirmed that these epitopes are naturally processed and functionally relevant. The design of more effective and less immunogenic AAV vectors, and precise immune monitoring of vector-infused subjects, are facilitated by these findings.

Bug bites and stings: When to see a dermatologist

Medline Plus

Full Text Available ... Mohs AUC MyDermPath+ Psoriasis Patient education resources Practice Management Center Coding and reimbursement Coding MACRA Fee schedule ... it’s important to take steps to reduce your risk. To help prevent bug bites, dermatologists recommend the ...

Bug bites and stings: When to see a dermatologist

Medline Plus

Full Text Available ... dress appropriately to prevent bug bites. Cover exposed skin as much as possible by wearing long-sleeved shirts, pants, socks and closed shoes instead of sandals. For additional protection, pull your socks up over your pants and ...

Electrostatic potential of human immunodeficiency virus type 2 and rhesus macaque simian immunodeficiency virus capsid proteins

Directory of Open Access Journals (Sweden)

Katarzyna eBozek

2012-06-01

Full Text Available Human immunodeficiency virus type 2 (HIV-2 and simian immunodeficiency virus isolated from a macaque monkey (SIVmac are assumed to have originated from simian immunodeficiency virus isolated from sooty mangabey (SIVsm. Despite their close similarity in genome structure, HIV-2 and SIVmac show different sensitivities to TRIM5α, a host restriction factor against retroviruses. The replication of HIV-2 strains is potently restricted by rhesus (Rh monkey TRIM5α, while that of SIVmac strain 239 (SIVmac239 is not. Viral capsid protein is the determinant of this differential sensitivity to TRIM5α, as the HIV-2 mutant carrying SIVmac239 capsid protein evaded Rh TRIM5α-mediated restriction. However, the molecular determinants of this restriction mechanism are unknown. Electrostatic potential on the protein-binding site is one of the properties regulating protein-protein interactions. In this study, we investigated the electrostatic potential on the interaction surface of capsid protein of HIV-2 strain GH123 and SIVmac239. Although HIV-2 GH123 and SIVmac239 capsid proteins share more than 87% amino acid identity, we observed a large difference between the two molecules with the HIV-2 GH123 molecule having predominantly positive and SIVmac239 predominantly negative electrostatic potential on the surface of the loop between α-helices 4 and 5 (L4/5. As L4/5 is one of the major determinants of Rh TRIM5α sensitivity of these viruses, the present results suggest that the binding site of the Rh TRIM5α may show complementarity to the HIV-2 GH123 capsid surface charge distribution.

Melt With This Kiss: Paralyzing and Liquefying Venom of The Assassin Bug Pristhesancus plagipennis (Hemiptera: Reduviidae).

Science.gov (United States)

Walker, Andrew A; Madio, Bruno; Jin, Jiayi; Undheim, Eivind A B; Fry, Bryan G; King, Glenn F

2017-04-01

Assassin bugs (Hemiptera: Heteroptera: Reduviidae) are venomous insects, most of which prey on invertebrates. Assassin bug venom has features in common with venoms from other animals, such as paralyzing and lethal activity when injected, and a molecular composition that includes disulfide-rich peptide neurotoxins. Uniquely, this venom also has strong liquefying activity that has been hypothesized to facilitate feeding through the narrow channel of the proboscis-a structure inherited from sap- and phloem-feeding phytophagous hemipterans and adapted during the evolution of Heteroptera into a fang and feeding structure. However, further understanding of the function of assassin bug venom is impeded by the lack of proteomic studies detailing its molecular composition.By using a combined transcriptomic/proteomic approach, we show that the venom proteome of the harpactorine assassin bug Pristhesancus plagipennis includes a complex suite of >100 proteins comprising disulfide-rich peptides, CUB domain proteins, cystatins, putative cytolytic toxins, triabin-like protein, odorant-binding protein, S1 proteases, catabolic enzymes, putative nutrient-binding proteins, plus eight families of proteins without homology to characterized proteins. S1 proteases, CUB domain proteins, putative cytolytic toxins, and other novel proteins in the 10-16-kDa mass range, were the most abundant venom components. Thus, in addition to putative neurotoxins, assassin bug venom includes a high proportion of enzymatic and cytolytic venom components likely to be well suited to tissue liquefaction. Our results also provide insight into the trophic switch to blood-feeding by the kissing bugs (Reduviidae: Triatominae). Although some protein families such as triabins occur in the venoms of both predaceous and blood-feeding reduviids, the composition of venoms produced by these two groups is revealed to differ markedly. These results provide insights into the venom evolution in the insect suborder

Formal solution for the fields within a beam-bug calibrator

International Nuclear Information System (INIS)

Fessenden, T J.

1998-01-01

For some time I was bothered by the fact that measurements of offsets in the various bug calibration setups never agreed with the simple formulae (2) used for determining electron beam position in the Livermore induction linacs and transport systems. About 1983 I realized that the discrepancy arises from the way the bug calibrator simulates an electron beam in a conducting pipe. At that time I solved the problem using the method presented here. Unfortunately, I did not write it up at that time. After considerable effort, I was able to repeat the calculation. Since I have little confidence that after a few years I could ever do it again, I felt obliged to write it up in some detail. Our beam bug calibrator consists of two conducting cylinders, nominally concentric, that simulate the electron beam within a drift tube. The radii of the larger cylinder is 2.3 times that of the smaller giving an electrical impedance of 50 Ohms to the coaxial combination. To simulate a beam off-axis within a drift tube, the inner tube is moved relative to the outer tube. This only approximately simulates the motion of a beam because the surface current on the inner tube redistributes in response to the translation. Fortunately, the fields of the translated inner cylinder can be found exactly using complex variable theory (1)

Solid-to-fluid DNA transition inside HSV-1 capsid close to the temperature of infection

Energy Technology Data Exchange (ETDEWEB)

Sae-Ueng, Udom; Li, Dong; Zuo, Xiaobing; Huffman, Jamie B.; Homa, Fred L.; Rau, Donald; Evilevitch, Alex

2014-10-01

DNA in the human Herpes simplex virus type 1 (HSV-1) capsid is packaged to a tight density. This leads to tens of atmospheres of internal pressure responsible for the delivery of the herpes genome into the cell nucleus. In this study we show that, despite its liquid crystalline state inside the capsid, the DNA is fluid-like, which facilitates its ejection into the cell nucleus during infection. We found that the sliding friction between closely packaged DNA strands, caused by interstrand repulsive interactions, is reduced by the ionic environment of epithelial cells and neurons susceptible to herpes infection. However, variations in the ionic conditions corresponding to neuronal activity can restrict DNA mobility in the capsid, making it more solid-like. This can inhibit intranuclear DNA release and interfere with viral replication. In addition, the temperature of the human host (37 °C) induces a disordering transition of the encapsidated herpes genome, which reduces interstrand interactions and provides genome mobility required for infection.

Chronic hepatitis B infection and HBV DNA-containing capsids: Modeling and analysis

Science.gov (United States)

Manna, Kalyan; Chakrabarty, Siddhartha P.

2015-05-01

We analyze the dynamics of chronic HBV infection taking into account both uninfected and infected hepatocytes along with the intracellular HBV DNA-containing capsids and the virions. While previous HBV models have included either the uninfected hepatocytes or the intracellular HBV DNA-containing capsids, our model accounts for both these two populations. We prove the conditions for local and global stability of both the uninfected and infected steady states in terms of the basic reproduction number. Further, we incorporate a time lag in the model to encompass the intracellular delay in the production of the infected hepatocytes and find that this delay does not affect the overall dynamics of the system. The results for the model and the delay model are finally numerically illustrated.

α-Defensin HD5 Inhibits Human Papillomavirus 16 Infection via Capsid Stabilization and Redirection to the Lysosome

Directory of Open Access Journals (Sweden)

Mayim E. Wiens

2017-01-01

Full Text Available α-Defensins are an important class of abundant innate immune effectors that are potently antiviral against a number of nonenveloped viral pathogens; however, a common mechanism to explain their ability to block infection by these unrelated viruses is lacking. We previously found that human defensin 5 (HD5 blocks a critical host-mediated proteolytic processing step required for human papillomavirus (HPV infection. Here, we show that bypassing the requirement for this cleavage failed to abrogate HD5 inhibition. Instead, HD5 altered HPV trafficking in the cell. In the presence of an inhibitory concentration of HD5, HPV was internalized and reached the early endosome. The internalized capsid became permeable to antibodies and proteases; however, HD5 prevented dissociation of the viral capsid from the genome, reduced viral trafficking to the trans-Golgi network, redirected the incoming viral particle to the lysosome, and accelerated the degradation of internalized capsid proteins. This mechanism is equivalent to the mechanism by which HD5 inhibits human adenovirus. Thus, our data support capsid stabilization and redirection to the lysosome during infection as a general antiviral mechanism of α-defensins against nonenveloped viruses.

Venture from the Interior-Herpesvirus pUL31 Escorts Capsids from Nucleoplasmic Replication Compartments to Sites of Primary Envelopment at the Inner Nuclear Membrane.

Science.gov (United States)

Bailer, Susanne M.

2017-11-25

Herpesviral capsid assembly is initiated in the nucleoplasm of the infected cell. Size constraints require that newly formed viral nucleocapsids leave the nucleus by an evolutionarily conserved vescular transport mechanism called nuclear egress. Mature capsids released from the nucleoplasm are engaged in a membrane-mediated budding process, composed of primary envelopment at the inner nuclear membrane and de-envelopment at the outer nuclear membrane. Once in the cytoplasm, the capsids receive their secondary envelope for maturation into infectious virions. Two viral proteins conserved throughout the herpesvirus family, the integral membrane protein pUL34 and the phosphoprotein pUL31, form the nuclear egress complex required for capsid transport from the infected nucleus to the cytoplasm. Formation of the nuclear egress complex results in budding of membrane vesicles revealing its function as minimal virus-encoded membrane budding and scission machinery. The recent structural analysis unraveled details of the heterodimeric nuclear egress complex and the hexagonal coat it forms at the inside of budding vesicles to drive primary envelopment. With this review, I would like to present the capsid-escort-model where pUL31 associates with capsids in nucleoplasmic replication compartments for escort to sites of primary envelopment thereby coupling capsid maturation and nuclear egress.

Computer games to teach hygiene: an evaluation of the e-Bug junior game.

Science.gov (United States)

Farrell, David; Kostkova, Patty; Weinberg, Julius; Lazareck, Lisa; Weerasinghe, Dasun; Lecky, Donna M; McNulty, Cliodna A M

2011-06-01

Handwashing, respiratory hygiene and antibiotic resistance remain major public health concerns. In order to facilitate an effective outcome when teaching the basic principles of hand and respiratory hygiene, educational interventions should first target school children. As computer games are ubiquitous in most children's lives, e-Bug developed computer games targeted at teaching children handwashing, respiratory hygiene and antibiotic resistance. The games were designed for two target audiences: junior school children (9-12 year olds); and senior school children (13-15 year olds). Between May and August 2009, the finalized junior game underwent an evaluation in three UK schools (in Glasgow, Gloucester and London), involving 62 children in the schools and ∼ 1700 players accessing the junior game online. The e-Bug junior game consists of a number of levels of play, each of which promotes a set of learning outcomes (LOs). These LOs, complementary to those in the e-Bug packs, are expressed through the game mechanics (the rules of the game) rather than through story or dialogue. Although the junior game's evaluation demonstrated a statistically significant change in the knowledge for only a small number of given LOs, because many children had the required knowledge already before playing the game, this is e-Bug's first statistical study on the junior game and the first comprehensive evaluation of its kind. Future work includes a re-examination of the quiz-style questionnaires utilized in this study and an exploration of the potential knowledge change acquired strictly through engagement.

Single and multiple in-season measurements as indicators of at-harvest cotton boll damage caused by verde plant bug (Hemiptera: Miridae).

Science.gov (United States)

Brewer, Michael J; Armstrong, J Scott; Parker, Roy D

2013-06-01

The ability to monitor verde plant bug, Creontiades signatus Distant (Hemiptera: Miridae), and the progression of cotton, Gossypium hirsutum L., boll responses to feeding and associated cotton boll rot provided opportunity to assess if single in-season measurements had value in evaluating at-harvest damage to bolls and if multiple in-season measurements enhanced their combined use. One in-season verde plant bug density measurement, three in-season plant injury measurements, and two at-harvest damage measurements were taken in 15 cotton fields in South Texas, 2010. Linear regression selected two measurements as potentially useful indicators of at-harvest damage: verde plant bug density (adjusted r2 = 0.68; P = 0.0004) and internal boll injury of the carpel wall (adjusted r2 = 0.72; P = 0.004). Considering use of multiple measurements, a stepwise multiple regression of the four in-season measurements selected a univariate model (verde plant bug density) using a 0.15 selection criterion (adjusted r2 = 0.74; P = 0.0002) and a bivariate model (verde plant bug density-internal boll injury) using a 0.25 selection criterion (adjusted r2 = 0.76; P = 0.0007) as indicators of at-harvest damage. In a validation using cultivar and water regime treatments experiencing low verde plant bug pressure in 2011 and 2012, the bivariate model performed better than models using verde plant bug density or internal boll injury separately. Overall, verde plant bug damaging cotton bolls exemplified the benefits of using multiple in-season measurements in pest monitoring programs, under the challenging situation when at-harvest damage results from a sequence of plant responses initiated by in-season insect feeding.

American Trypanosomiasis (Also Known as Chagas Disease) Triatomine Bug FAQs

Science.gov (United States)

... are typically found in the southern United States, Mexico, Central America, and South America (as far south ... feed on the blood of mammals (including humans), birds, and reptiles. Triatomine bugs live in a wide ...

Spectral sideband produced by a hemispherical concave multilayer on the African shield-bug Calidea panaethiopica (Scutelleridae)

Science.gov (United States)

Vigneron, Jean Pol; Ouedraogo, Moussa; Colomer, Jean-François; Rassart, Marie

2009-02-01

The African shield-backed bug Calidea panaethiopica is a very colorful insect which produces a range of iridescent yellow, green, and blue reflections. The cuticle of the dorsal side of the insect, on the shield, the prothorax and part of the head, is pricked of uniformly distributed hemispherical hollow cavities a few tens micrometers deep. Under normal illumination and viewing the insect’s muffin-tin shaped surface gives rise to two distinct colors: a yellow spot arising from the bottom of the well and a blue annular cloud that appears to float around the yellow spot. This effect is explained by multiple reflections on a hemispherical Bragg mirror with a mesoscopic curvature. A multiscale computing methodology was found to be needed to evaluate the reflection spectrum for such a curved multilayer. This multiscale approach is very general and should be useful for dealing with visual effects in many natural and artificial systems.

Dengue Virus Uses a Non-Canonical Function of the Host GBF1-Arf-COPI System for Capsid Protein Accumulation on Lipid Droplets.

Science.gov (United States)

Iglesias, Nestor G; Mondotte, Juan A; Byk, Laura A; De Maio, Federico A; Samsa, Marcelo M; Alvarez, Cecilia; Gamarnik, Andrea V

2015-09-01

Dengue viruses cause the most important human viral disease transmitted by mosquitoes. In recent years, a great deal has been learned about molecular details of dengue virus genome replication; however, little is known about genome encapsidation and the functions of the viral capsid protein. During infection, dengue virus capsid progressively accumulates around lipid droplets (LDs) by an unknown mechanism. Here, we examined the process by which the viral capsid is transported from the endoplasmic reticulum (ER) membrane, where the protein is synthesized, to LDs. Using different methods of intervention, we found that the GBF1-Arf1/Arf4-COPI pathway is necessary for capsid transport to LDs, while the process is independent of both COPII components and Golgi integrity. The transport was sensitive to Brefeldin A, while a drug resistant form of GBF1 was sufficient to restore capsid subcellular distribution in infected cells. The mechanism by which LDs gain or lose proteins is still an open question. Our results support a model in which the virus uses a non-canonical function of the COPI system for capsid accumulation on LDs, providing new ideas for antiviral strategies. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Structure of a Spumaretrovirus Gag Central Domain Reveals an Ancient Retroviral Capsid.

Directory of Open Access Journals (Sweden)

Neil J Ball

2016-11-01

Full Text Available The Spumaretrovirinae, or foamy viruses (FVs are complex retroviruses that infect many species of monkey and ape. Despite little sequence homology, FV and orthoretroviral Gag proteins perform equivalent functions, including genome packaging, virion assembly, trafficking and membrane targeting. However, there is a paucity of structural information for FVs and it is unclear how disparate FV and orthoretroviral Gag molecules share the same function. To probe the functional overlap of FV and orthoretroviral Gag we have determined the structure of a central region of Gag from the Prototype FV (PFV. The structure comprises two all α-helical domains NtDCEN and CtDCEN that although they have no sequence similarity, we show they share the same core fold as the N- (NtDCA and C-terminal domains (CtDCA of archetypal orthoretroviral capsid protein (CA. Moreover, structural comparisons with orthoretroviral CA align PFV NtDCEN and CtDCEN with NtDCA and CtDCA respectively. Further in vitro and functional virological assays reveal that residues making inter-domain NtDCEN-CtDCEN interactions are required for PFV capsid assembly and that intact capsid is required for PFV reverse transcription. These data provide the first information that relates the Gag proteins of Spuma and Orthoretrovirinae and suggests a common ancestor for both lineages containing an ancient CA fold.

Structure of a Spumaretrovirus Gag Central Domain Reveals an Ancient Retroviral Capsid

Science.gov (United States)

Dutta, Moumita; Pollard, Dominic J.; Goldstone, David C.; Ramos, Andres; Müllers, Erik; Stirnnagel, Kristin; Stanke, Nicole; Lindemann, Dirk; Taylor, William R.; Rosenthal, Peter B.

2016-01-01

The Spumaretrovirinae, or foamy viruses (FVs) are complex retroviruses that infect many species of monkey and ape. Despite little sequence homology, FV and orthoretroviral Gag proteins perform equivalent functions, including genome packaging, virion assembly, trafficking and membrane targeting. However, there is a paucity of structural information for FVs and it is unclear how disparate FV and orthoretroviral Gag molecules share the same function. To probe the functional overlap of FV and orthoretroviral Gag we have determined the structure of a central region of Gag from the Prototype FV (PFV). The structure comprises two all α-helical domains NtDCEN and CtDCEN that although they have no sequence similarity, we show they share the same core fold as the N- (NtDCA) and C-terminal domains (CtDCA) of archetypal orthoretroviral capsid protein (CA). Moreover, structural comparisons with orthoretroviral CA align PFV NtDCEN and CtDCEN with NtDCA and CtDCA respectively. Further in vitro and functional virological assays reveal that residues making inter-domain NtDCEN—CtDCEN interactions are required for PFV capsid assembly and that intact capsid is required for PFV reverse transcription. These data provide the first information that relates the Gag proteins of Spuma and Orthoretrovirinae and suggests a common ancestor for both lineages containing an ancient CA fold. PMID:27829070

Bullous reactions to bed bug bites reflect cutaneous vasculitis

Science.gov (United States)

This study evaluates bullous cutaneous reactions and sequential histopathology in an individual sensitized to bed bug bites in an effort to better understand the allergic response and histology associated with these bites. There was a progression of the inflammatory response across time ranging from...

Specific Inhibitors of HIV Capsid Assembly Binding to the C-Terminal Domain of the Capsid Protein: Evaluation of 2-Arylquinazolines as Potential Antiviral Compounds

Czech Academy of Sciences Publication Activity Database

Machara, A.; Lux, V.; Kožíšek, Milan; Grantz Šašková, Klára; Štěpánek, O.; Kotora, M.; Parkan, Kamil; Pávová, Marcela; Glass, B.; Sehr, P.; Lewis, J.; Müller, B.; Kräusslich, H. G.; Konvalinka, Jan

2016-01-01

Roč. 59, č. 2 (2016), s. 545-558 ISSN 0022-2623 R&D Projects: GA ČR GA13-19561S EU Projects: European Commission(XE) 201095 - HIV ACE Institutional support: RVO:61388963 Keywords : HIV -1 assembly * capsid * high-throughput screening * AlphaScreen assay Subject RIV: CE - Biochemistry Impact factor: 6.259, year: 2016

Virtual Reality Check: Teachers Use Bug-in-Ear Coaching to Practice Feedback Techniques with Student Avatars

Science.gov (United States)

Elford, Marti; Carter, Richard A., Jr.; Aronin, Sara

2013-01-01

There is not just one way to give feedback, nor is there just one kind of feedback. Bug-in-ear technology, which allows coaches to give teachers in the classroom immediate feedback, has been used successfully for 35 years. In an updated twist on this method, researchers at the University of Kansas used bug-in-ear coaching in a virtual classroom…

Comparative susceptibilities of different life stages of the tarnished plant bug (Hemiptera: miridae) to three classes of insecticide

Science.gov (United States)

Insecticidal control of the tarnished plant bug, Lygus lineolaris (Palisot de Beauvois), is targeted at the adult and nymphal stages, but there is little information on relative susceptibilities of these insects to insecticides. Tarnished plant bug adults were collected from various locations in Mi...

Climbing ability of teneral and sclerotized adult bed bugs and assessment of adhesive properties of the exoskeletal fluid using atomic force microscopy.

Directory of Open Access Journals (Sweden)

Kevin R Hinson

Full Text Available We observed that teneral adults (72 h, and sclerotized group 2 (S2 bed bugs (n = 30, height climbed = 2.64 cm at >72 h post molt. When heights from all climbing events were summed, teneral bed bugs (650.8 cm climbed differed significantly (P< 0.01 from recently sclerotized (82 cm climbed and sclerotized (group 1 = 104.6 cm climbed, group 2 = 107.8 cm climbed bed bugs. These findings suggested that the external surface of teneral bed bug exoskeletons possess an adhesive property. Using atomic force microscopy (AFM, we found that adhesion force of an exoskeletal (presumably molting fluid decreased almost five-fold from 88 to 17 nN within an hour of molting. Our findings may have implications for laboratory safety and the effectiveness of bed bug traps, barriers, and biomimetic-based adhesives.

Development and validation of novel AAV2 random libraries displaying peptides of diverse lengths and at diverse capsid positions.

Science.gov (United States)

Naumer, Matthias; Ying, Ying; Michelfelder, Stefan; Reuter, Antje; Trepel, Martin; Müller, Oliver J; Kleinschmidt, Jürgen A

2012-05-01

Libraries based on the insertion of random peptide ligands into the capsid of adeno-associated virus type 2 (AAV2) have been widely used to improve the efficiency and selectivity of the AAV vector system. However, so far only libraries of 7-mer peptide ligands have been inserted at one well-characterized capsid position. Here, we expanded the combinatorial AAV2 display system to a panel of novel AAV libraries, displaying peptides of 5, 7, 12, 19, or 26 amino acids in length at capsid position 588 or displaying 7-mer peptides at position 453, the most prominently exposed region of the viral capsid. Library selections on two unrelated cell types-human coronary artery endothelial cells and rat cardiomyoblasts-revealed the isolation of cell type-characteristic peptides of different lengths mediating strongly improved target-cell transduction, except for the 26-mer peptide ligands. Characterization of vector selectivity by transduction of nontarget cells and comparative gene-transduction analysis using a panel of 44 human tumor cell lines revealed that insertion of different-length peptides allows targeting of distinct cellular receptors for cell entry with similar efficiency, but with different selectivity. The application of such novel AAV2 libraries broadens the spectrum of targetable receptors by capsid-modified AAV vectors and provides the opportunity to choose the best suited targeting ligand for a certain application from a number of different candidates.

Seasonal and regional dynamics of M. ulcerans transmission in environmental context: deciphering the role of water bugs as hosts and vectors.

Science.gov (United States)

Marion, Estelle; Eyangoh, Sara; Yeramian, Edouard; Doannio, Julien; Landier, Jordi; Aubry, Jacques; Fontanet, Arnaud; Rogier, Christophe; Cassisa, Viviane; Cottin, Jane; Marot, Agnès; Eveillard, Matthieu; Kamdem, Yannick; Legras, Pierre; Deshayes, Caroline; Saint-André, Jean-Paul; Marsollier, Laurent

2010-07-06

Buruli ulcer, the third mycobacterial disease after tuberculosis and leprosy, is caused by the environmental mycobacterium M. ulcerans. Various modes of transmission have been suspected for this disease, with no general consensus acceptance for any of them up to now. Since laboratory models demonstrated the ability of water bugs to transmit M. ulcerans, a particular attention is focused on the transmission of the bacilli by water bugs as hosts and vectors. However, it is only through detailed knowledge of the biodiversity and ecology of water bugs that the importance of this mode of transmission can be fully assessed. It is the objective of the work here to decipher the role of water bugs in M. ulcerans ecology and transmission, based on large-scale field studies. The distribution of M. ulcerans-hosting water bugs was monitored on previously unprecedented time and space scales: a total of 7,407 water bugs, belonging to large number of different families, were collected over one year, in Buruli ulcer endemic and non endemic areas in central Cameroon. This study demonstrated the presence of M. ulcerans in insect saliva. In addition, the field results provided a full picture of the ecology of transmission in terms of biodiversity and detailed specification of seasonal and regional dynamics, with large temporal heterogeneity in the insect tissue colonization rate and detection of M. ulcerans only in water bug tissues collected in Buruli ulcer endemic areas. The large-scale detection of bacilli in saliva of biting water bugs gives enhanced weight to their role in M. ulcerans transmission. On practical grounds, beyond the ecological interest, the results concerning seasonal and regional dynamics can provide an efficient tool in the hands of sanitary authorities to monitor environmental risks associated with Buruli ulcer.

Bug bites and stings: When to see a dermatologist

Medline Plus

Full Text Available ... Private payer Medicare physician payment MACRA implementation Alternative payment models Fee schedule State policy State policy and action ... a dermatologist Why see a board-certified dermatologist? Home Public and patients Skin, hair, and nail care Injured skin Bug bites and stings "); (function () { var ...

Biochemical Assay Detects Feeding Damage to Loblolly Pine Seeds Caused by the Leaffooted Pine Seed Bug (Hemiptera: Coreidae)

Science.gov (United States)

Cameron G. Lait; Daniel R. Miller; Sarah L. Bates; John H. Borden; Allison R. Kermode

2003-01-01

A large number of proteins in salivary gland extracts of the leaffooted pine seed bug, Leptoglossus corculus Say, were strongly recognized by a polyclonal antibody-based assay developed for detecting saliva of the western conifer seed bug, Lepfoglossus occidentalis Heidemann, in lodgepole pine, Pinus contorta var...

Outer capsid proteins induce the formation of pores in epithelial cells

International Nuclear Information System (INIS)

Ruiz, M; Abad M; Michelangely, F; Charpilienne, A; Cohen, J

1995-01-01

Two mechanisms of entrance in cell of the rotavirus, during the infection, were proposed: a direct entrance through the plasmatic membrane or by means of endocytosis. In the two cases, a permeabilization mechanism of the membrane (cellular or of the endocytic vesicle, respectively) should occur. It has been shown that the rotavirus induces permeabilization of liposomes and of membrane vesicles. In this work, are studied the changes of intact cells permeability, measuring the entrance of e tide bromides. Viral particles of double capsid of the RF stump produce an increase of the cells membrane MA104 permeability, while the simple capsid ones don't induce effect. This phenomenon requires the particles trypsinization, and occurs in a means where the concentration of free Ca is lower to 1 micromolar. The temporary course of the fluorescence increase is sigmoid. The latency, the speed and the width depend on the relationship of virus / cell, and it can be observed up to 100% of permeabilization in relation to the effect of digitonin. The pores induced in the membrane by the rotavirus are irreversible. The permeabilizer effect of the rotavirus on the membrane was observed in other cellular lines as Hela and HT29, but not in the L929 ones. These results suggest that one or more proteins of the external capsid are responsible s of the effect. These could be involved in the penetration process of the virus towards the cytoplasm and could be one of the restrictive factor of the cell infection by means of the virus [es

The Bad Bugs Book Club: Science, Literacy, and Engagement

Directory of Open Access Journals (Sweden)

Joanna Verran

2013-01-01

Full Text Available The Bad Bugs Bookclub was launched in 2009. It comprises scientists and non-scientists. The aim of the Bookclub is to read and discuss novels where infectious disease forms part of the plot, in order to enhance learning about microbiology. The focus of the discussion is on appreciation of the novel, its scientific (microbiologic accuracy and relevance to contemporary microbiology. There are several potential audiences for the Bad Bugs Bookclub, for example students in a classroom setting, or in a more social environment, and/or the general public.  Meeting reports and reading guides have been posted on a dedicated website. For education purposes, additional project work for assessment is suggested for students reading each novel. Bookclub meetings may be held on particular dates in the microbiologic calendar, coupled with additional public engagement activities and student participation. The approach has significant flexibility in terms of intended audience, assessment and extension work, and provides a refreshing and stimulating alternative means for talking about microbiology.

Foot-and-mouth disease virus capsid proteins; analysis of protein processing, assembly and utility as vaccines

DEFF Research Database (Denmark)

Belsham, Graham

Foot-and-mouth disease (FMD) remains one of the most economically important infectious diseases of production animals globally. The infection is caused by foot-and-mouth disease virus (FMDV), a member of the picornavirus family. The positive sense RNA genome of the virus includes a single, large......, open reading frame that encodes a polyprotein. The intact polyprotein is never observed as it is processed, both during and after translation, to 15 different mature proteins plus a variety of precursors. The FMDV capsid protein precursor, P1-2A, is cleaved by the virus encoded 3C protease (3Cpro......) to generate VP0, VP3, VP1 and the peptide 2A. Sixty copies of each of the capsid proteins “self-assemble” into empty capsid particles or with the RNA genome into infectious viruses. These particles normally lack 2A but it is possible to construct and isolate mutant FMDVs in which the cleavage of the VP1/2A...

Verde plant bug associatioin with boll damage including cotton boll rot and potential in-season indicators of damage

Science.gov (United States)

Cotton along the Gulf Coast of south Texas has experienced loss from cotton boll rot especially during the last 10 to 15 years, and stink bugs and plant bugs (Hemiptera: Pentatomidae and Miridae) that feed on cotton bolls have been suspected in introducing the disease. A replicated grower field surv...

Retargeting of rat parvovirus H-1PV to cancer cells through genetic engineering of the viral capsid.

Science.gov (United States)

Allaume, Xavier; El-Andaloussi, Nazim; Leuchs, Barbara; Bonifati, Serena; Kulkarni, Amit; Marttila, Tiina; Kaufmann, Johanna K; Nettelbeck, Dirk M; Kleinschmidt, Jürgen; Rommelaere, Jean; Marchini, Antonio

2012-04-01

The rat parvovirus H-1PV is a promising anticancer agent given its oncosuppressive properties and the absence of known side effects in humans. H-1PV replicates preferentially in transformed cells, but the virus can enter both normal and cancer cells. Uptake by normal cells sequesters a significant portion of the administered viral dose away from the tumor target. Hence, targeting H-1PV entry specifically to tumor cells is important to increase the efficacy of parvovirus-based treatments. In this study, we first found that sialic acid plays a key role in H-1PV entry. We then genetically engineered the H-1PV capsid to improve its affinity for human tumor cells. By analogy with the resolved crystal structure of the closely related parvovirus minute virus of mice, we developed an in silico three-dimensional (3D) model of the H-1PV wild-type capsid. Based on this model, we identified putative amino acids involved in cell membrane recognition and virus entry at the level of the 2-fold axis of symmetry of the capsid, within the so-called dimple region. In situ mutagenesis of these residues significantly reduced the binding and entry of H-1PV into permissive cells. We then engineered an entry-deficient viral capsid and inserted a cyclic RGD-4C peptide at the level of its 3-fold axis spike. This peptide binds α(v)β(3) and α(v)β(5) integrins, which are overexpressed in cancer cells and growing blood vessels. The insertion of the peptide rescued viral infectivity toward cells overexpressing α(v)β(5) integrins, resulting in the efficient killing of these cells by the reengineered virus. This work demonstrates that H-1PV can be genetically retargeted through the modification of its capsid, showing great promise for a more efficient use of this virus in cancer therapy.

Insecticide assays against the brown stink bug feeding on pecan

Science.gov (United States)

The brown stink bug, Euschistus servus (Say) (Hemiptera: Pentatomidae), is an economic pest of pecan, Carya illinoinensis (Wangenh) K. Koch (Juglandaceae), and other agronomic crops across the southeastern U.S. Management of this pest is mainly via insecticides. Many commercial products indicate o...

Respiratory function of the plastron in the aquatic bug Aphelocheirus aestivalis (Hemiptera, Aphelocheiridae).

Science.gov (United States)

Seymour, Roger S; Jones, Karl K; Hetz, Stefan K

2015-09-01

The river bug Aphelocheirus aestivalis is a 40 mg aquatic insect that, as an adult, relies totally on an incompressible physical gill to exchange respiratory gases with the water. The gill (called a 'plastron') consists of a stationary layer of air held in place on the body surface by millions of tiny hairs that support a permanent air-water interface, so that the insect never has to renew the gas at the water's surface. The volume of air in the plastron is extremely small (0.14 mm(3)), under slightly negative pressure and connected to the gas-filled tracheal system through spiracles on the cuticle. Here, we measure PO2 of the water and within the plastron gas with O2-sensing fibre optics to understand the effectiveness and limitations of the gas exchanger. The difference in PO2 is highest in stagnant water and decreases with increasing convection over the surface. Respiration of bugs in water-filled vials varies between 33 and 296 pmol O2 s(-1), depending on swimming activity. The effective thickness of the boundary layer around the plastron was calculated from respiration rate, PO2 difference and plastron surface area, according to the Fick diffusion equation and verified by direct measurements with the fibre-optic probes. In stagnant water, the boundary layer is approximately 500 μm thick, which nevertheless can satisfy the demands of resting bugs, even if the PO2 of the free water decreases to half that of air saturation. Active bugs require thinner boundary layers (∼ 100 μm), which are achieved by living in moving water or by swimming. © 2015. Published by The Company of Biologists Ltd.

The Dual Role of an ESCRT-0 Component HGS in HBV Transcription and Naked Capsid Secretion.

Directory of Open Access Journals (Sweden)

Shu-Fan Chou

2015-10-01

Full Text Available The Endosomal Sorting Complex Required for Transport (ESCRT is an important cellular machinery for the sorting and trafficking of ubiquitinated cargos. It is also known that ESCRT is required for the egress of a number of viruses. To investigate the relationship between ESCRT and hepatitis B virus (HBV, we conducted an siRNA screening of ESCRT components for their potential effect on HBV replication and virion release. We identified a number of ESCRT factors required for HBV replication, and focused our study here on HGS (HRS, hepatocyte growth factor-regulated tyrosine kinase substrate in the ESCRT-0 complex. Aberrant levels of HGS suppressed HBV transcription, replication and virion secretion. Hydrodynamic delivery of HGS in a mouse model significantly suppressed viral replication in the liver and virion secretion in the serum. Surprisingly, overexpression of HGS stimulated the release of HBV naked capsids, irrespective of their viral RNA, DNA, or empty contents. Mutant core protein (HBc 1-147 containing no arginine-rich domain (ARD failed to secrete empty virions with or without HGS. In contrast, empty naked capsids of HBc 1-147 could still be promoted for secretion by HGS. HGS exerted a strong positive effect on the secretion of naked capsids, at the expense of a reduced level of virions. The association between HGS and HBc appears to be ubiquitin-independent. Furthermore, HBc is preferentially co-localized with HGS near the cell periphery, instead of near the punctate endosomes in the cytoplasm. In summary, our work demonstrated the importance of an optimum level of HGS in HBV propagation. In addition to an effect on HBV transcription, HGS can diminish the pool size of intracellular nucleocapsids with ongoing genome maturation, probably in part by promoting the secretion of naked capsids. The secretion routes of HBV virions and naked capsids can be clearly distinguished based on the pleiotropic effect of HGS involved in the ESCRT-0 complex.

The Dual Role of an ESCRT-0 Component HGS in HBV Transcription and Naked Capsid Secretion

Science.gov (United States)

Chou, Shu-Fan; Tsai, Ming-Lin; Huang, Jyun-Yuan; Chang, Ya-Shu; Shih, Chiaho

2015-01-01

The Endosomal Sorting Complex Required for Transport (ESCRT) is an important cellular machinery for the sorting and trafficking of ubiquitinated cargos. It is also known that ESCRT is required for the egress of a number of viruses. To investigate the relationship between ESCRT and hepatitis B virus (HBV), we conducted an siRNA screening of ESCRT components for their potential effect on HBV replication and virion release. We identified a number of ESCRT factors required for HBV replication, and focused our study here on HGS (HRS, hepatocyte growth factor-regulated tyrosine kinase substrate) in the ESCRT-0 complex. Aberrant levels of HGS suppressed HBV transcription, replication and virion secretion. Hydrodynamic delivery of HGS in a mouse model significantly suppressed viral replication in the liver and virion secretion in the serum. Surprisingly, overexpression of HGS stimulated the release of HBV naked capsids, irrespective of their viral RNA, DNA, or empty contents. Mutant core protein (HBc 1–147) containing no arginine-rich domain (ARD) failed to secrete empty virions with or without HGS. In contrast, empty naked capsids of HBc 1–147 could still be promoted for secretion by HGS. HGS exerted a strong positive effect on the secretion of naked capsids, at the expense of a reduced level of virions. The association between HGS and HBc appears to be ubiquitin-independent. Furthermore, HBc is preferentially co-localized with HGS near the cell periphery, instead of near the punctate endosomes in the cytoplasm. In summary, our work demonstrated the importance of an optimum level of HGS in HBV propagation. In addition to an effect on HBV transcription, HGS can diminish the pool size of intracellular nucleocapsids with ongoing genome maturation, probably in part by promoting the secretion of naked capsids. The secretion routes of HBV virions and naked capsids can be clearly distinguished based on the pleiotropic effect of HGS involved in the ESCRT-0 complex. PMID

Scaffold expulsion and genome packaging trigger stabilization of herpes simplex virus capsids

NARCIS (Netherlands)

Roos, W.H.; Radtke, K.; Kniesmeijer, E.G.R.; Geertsema, H.J.; Sodeik, B.; Wuite, G.J.L.

2009-01-01

Herpes simplex virus type 1 (HSV1) capsids undergo extensive structural changes during maturation and DNA packaging. As a result, they become more stable and competent for nuclear egress. To further elucidate this stabilization process, we used biochemical and nanoindentation approaches to analyze

Scaffold expulsion and genome packaging trigger stabilization of Herpes Simplex Virus capsids

NARCIS (Netherlands)

Roos, W.H.; Radtke, K.; Kniesmeijer, E.; Geertsema, H.J.; Sodeik, B.; Wuite, G.J.L.

2009-01-01

Herpes simplex virus type 1 (HSV1) capsids undergo extensive structural changes during maturation and DNA packaging. As a result, they become more stable and competent for nuclear egress. To further elucidate this stabilization process, we used biochemical and nanoindentation approaches to analyze

Bug bites and stings: When to see a dermatologist

Medline Plus

Full Text Available ... 0; c public SPOT Skin Cancer™ Diseases and treatments Skin, hair, and nail care Skin care Hair care / hair loss Injured skin Blisters Bug bites and stings How to remove a tick How to treat a bee sting When to see a dermatologist Burns Frostbite Splinters Treating sunburn Wound care Nail care ...

Antibody Competition Reveals Surface Location of HPV L2 Minor Capsid Protein Residues 17-36.

Science.gov (United States)

Bywaters, Stephanie M; Brendle, Sarah A; Tossi, Kerstin P; Biryukov, Jennifer; Meyers, Craig; Christensen, Neil D

2017-11-10

The currently available nonavalent human papillomavirus (HPV) vaccine exploits the highly antigenic L1 major capsid protein to promote high-titer neutralizing antibodies, but is limited to the HPV types included in the vaccine since the responses are highly type-specific. The limited cross-protection offered by the L1 virus-like particle (VLP) vaccine warrants further investigation into cross-protective L2 epitopes. The L2 proteins are yet to be fully characterized as to their precise placement in the virion. Adding to the difficulties in localizing L2, studies have suggested that L2 epitopes are not well exposed on the surface of the mature capsid prior to cellular engagement. Using a series of competition assays between previously mapped anti-L1 monoclonal antibodies (mAbs) (H16.V5, H16.U4 and H16.7E) and novel anti-L2 mAbs, we probed the capsid surface for the location of an L2 epitope (aa17-36). The previously characterized L1 epitopes together with our competition data is consistent with a proposed L2 epitope within the canyons of pentavalent capsomers.

Insecticide susceptibility of Nezara viridula (Heteroptera: Pentatomidae) and three other stink bug species composing a soybean pest complex in Japan.

Science.gov (United States)

Takeuchi, Hiroaki; Endo, Nobuyuki

2012-06-01

The susceptibility of the stink bug species Nezara viridula (L.), Nezara antennata Scott, Piezodorus hybneri (Gmelin), and Riptortus pedestris (F.) to insecticides was tested, establishing their 50% lethal dose (LD50) values as baseline data. Third instars and adults of the four species were treated by topical application with seven insecticides: fenitrothion, fenthion, etofenprox, silafluofen, dinotefuran, clothianidin, and ethiprole. The weight of the stink bug and weight of the insecticide applied to each bug were used as explanatory variables in the probit regression analysis. The effect of the body weight on the dose-response relationship, the proportional model, was not uniform among the tested insecticide-stink bug combinations. However, the basic model fit all combinations and could estimate LD50 values successfully. Therefore, LD50 values at the medium (average) weight estimated by the basic model were selected to describe the susceptibility of the stink bugs. The LD50 value of silafluofen for N. viridula adults, and that of silafluofen and etofenprox for N. antennata adults, was at least 2,338 ng greater than the other species exposed to each insecticide. Almost all of the LD50 values for adults were over 10 times greater than those of the same species' nymphs treated with the same insecticide. Thus monitoring of occurring species and their developmental stages is important for controlling effectively the stink bug pest complex by insecticides, especially by silafluofen or etofenprox. The estimated LD50 values can be used as baseline data to compare the susceptibility of the species collected in another year or location.

Bed Bug (Cimex lectularius L. Population Composition as Determined by Baited Traps

Directory of Open Access Journals (Sweden)

Elizabeth J. Schaafsma

2012-04-01

Full Text Available Two established field populations of bed bugs were sampled using host-mimicking traps baited with a combination of CO2, heat and a synthetic kairomone. The proportion of first instar nymphs (between 52% and 78% of all captured insects was significantly higher than reported in previous studies, which had employed different sampling methods. The proportion of adults was correspondingly much lower than previously reported, between 5% and 7% of total capture. As many as 120 bed bugs were captured in a single trap in one night; the variation in catches between sampling locations within the same room and between days at the same location indicates that multiple nights of trapping may be required to obtain an accurate representation of population structure.

Characterization and Comparative Analysis of Olfactory Receptor Co-Receptor Orco Orthologs Among Five Mirid Bug Species

Directory of Open Access Journals (Sweden)

Qi Wang

2018-03-01

Full Text Available The phytophagous mirid bugs of Apolygus lucorum, Lygus pratensis as well as three Adelphocoris spp., including Adelphocoris lineolatus, A. suturalis, and A. fasciaticollis are major pests of multiple agricultural crops in China, which have distinct geographical distribution and occurrence ranges. Like many insect species, these bugs heavily rely on olfactory cues to search preferred host plants, thereby investigation on functional co-evolution and divergence of olfactory genes seems to be necessary and is of great interest. In the odorant detection pathway, olfactory receptor co-receptor (Orco plays critical role in the perception of odors. In this study, we identified the full-length cDNA sequences encoding three putative Orcos (AsutOrco, AfasOrco, and LpraOrco in bug species of A. suturalis, A. fasciaticollis, and L. pratensis based on homology cloning method. Next, sequence alignment, membrane topology and gene structure analysis showed that these three Orco orthologs together with previously reported AlinOrco and AlucOrco shared high amino acid identities and similar topology structure, but had different gene structure especially at the length and insertion sites of introns. Furthermore, the evolutional estimation on the ratios of non-synonymous to synonymous (Ka/Ks revealed that Orco genes were under strong purifying selection, but the degrees of variation were significant different between genera. The results of quantitative real-time PCR experiments showed that these five Orco genes had a similar antennae-biased tissue expression pattern. Taking these data together, it is thought that Orco genes in the mirid species could share conserved olfaction roles but had different evolution rates. These findings would lay a foundation to further investigate the molecular mechanisms of evolutionary interactions between mirid bugs and their host plants, which might in turn contribute to the development of pest management strategy for mirid bugs.

Biochemical and biophysical characterization of cell-free synthesized Rift Valley fever virus nucleoprotein capsids enables in vitro screening to identify novel antivirals.

Science.gov (United States)

Broce, Sean; Hensley, Lisa; Sato, Tomoharu; Lehrer-Graiwer, Joshua; Essrich, Christian; Edwards, Katie J; Pajda, Jacqueline; Davis, Christopher J; Bhadresh, Rami; Hurt, Clarence R; Freeman, Beverly; Lingappa, Vishwanath R; Kelleher, Colm A; Karpuj, Marcela V

2016-05-14

Viral capsid assembly involves the oligomerization of the capsid nucleoprotein (NP), which is an essential step in viral replication and may represent a potential antiviral target. An in vitro transcription-translation reaction using a wheat germ (WG) extract in combination with a sandwich ELISA assay has recently been used to identify small molecules with antiviral activity against the rabies virus. Here, we examined the application of this system to viruses with capsids with a different structure, such as the Rift Valley fever virus (RVFV), the etiological agent of a severe emerging infectious disease. The biochemical and immunological characterization of the in vitro-generated RVFV NP assembly products enabled the distinction between intermediately and highly ordered capsid structures. This distinction was used to establish a screening method for the identification of potential antiviral drugs for RVFV countermeasures. These results indicated that this unique analytical system, which combines nucleoprotein oligomerization with the specific immune recognition of a highly ordered capsid structure, can be extended to various viral families and used both to study the early stages of NP assembly and to assist in the identification of potential antiviral drugs in a cost-efficient manner. Reviewed by Jeffry Skolnick and Noah Isakov. For the full reviews please go to the Reviewers' comments section.

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.

Herpes Simplex Virus 1 Us3 Deletion Mutant is Infective Despite Impaired Capsid Translocation to the Cytoplasm

Directory of Open Access Journals (Sweden)

Peter Wild

2015-01-01

Full Text Available Herpes simplex virus 1 (HSV-1 capsids are assembled in the nucleus bud at the inner nuclear membrane into the perinuclear space, acquiring envelope and tegument. In theory, these virions are de-enveloped by fusion of the envelope with the outer nuclear membrane and re-enveloped by Golgi membranes to become infective. Us3 enables the nucleus to cytoplasm capsid translocation. Nevertheless, Us3 is not essential for the production of infective progeny viruses. Determination of phenotype distribution by quantitative electron microscopy, and calculation per mean nuclear or cell volume revealed the following: (i The number of R7041(∆US3 capsids budding at the inner nuclear membrane was significantly higher than that of wild type HSV-1; (ii The mean number of R7041(∆US3 virions per mean cell volume was 2726, that of HSV-1 virions 1460 by 24 h post inoculation; (iii 98% of R7041(∆US3 virions were in the perinuclear space; (iv The number of R7041(∆US3 capsids in the cytoplasm, including those budding at Golgi membranes, was significantly reduced. Cell associated R7041(∆US3 yields were 2.37 × 108 and HSV-1 yields 1.57 × 108 PFU/mL by 24 h post inoculation. We thus conclude that R7041(∆US3 virions, which acquire envelope and tegument by budding at the inner nuclear membrane into the perinuclear space, are infective.

RNA interference of NADPH-cytochrome P450 reductase results in reduced insecticide resistance in the bed bug, Cimex lectularius.

Science.gov (United States)

Zhu, Fang; Sams, Sarah; Moural, Tim; Haynes, Kenneth F; Potter, Michael F; Palli, Subba R

2012-01-01

NADPH-cytochrome P450 reductase (CPR) plays a central role in cytochrome P450 action. The genes coding for P450s are not yet fully identified in the bed bug, Cimex lectularius. Hence, we decided to clone cDNA and knockdown the expression of the gene coding for CPR which is suggested to be required for the function of all P450s to determine whether or not P450s are involved in resistance of bed bugs to insecticides. The full length Cimex lectularius CPR (ClCPR) cDNA was isolated from a deltamethrin resistant bed bug population (CIN-1) using a combined PCR strategy. Bioinformatics and in silico modeling were employed to identify three conserved binding domains (FMN, FAD, NADP), a FAD binding motif, and the catalytic residues. The critical amino acids involved in FMN, FAD, NADP binding and their putative functions were also analyzed. No signal peptide but a membrane anchor domain with 21 amino acids which facilitates the localization of ClCPR on the endoplasmic reticulum was identified in ClCPR protein. Phylogenetic analysis showed that ClCPR is closer to the CPR from the body louse, Pediculus humanus corporis than to the CPRs from the other insect species studied. The ClCPR gene was ubiquitously expressed in all tissues tested but showed an increase in expression as immature stages develop into adults. We exploited the traumatic insemination mechanism of bed bugs to inject dsRNA and successfully knockdown the expression of the gene coding for ClCPR. Suppression of the ClCPR expression increased susceptibility to deltamethrin in resistant populations but not in the susceptible population of bed bugs. These data suggest that P450-mediated metabolic detoxification may serve as one of the resistance mechanisms in bed bugs.

Aflatoxin Contamination Detected in Nutrient and Anti-Oxidant Rich Edible Stink Bug Stored in Recycled Grain Containers.

Directory of Open Access Journals (Sweden)

Robert Musundire

Full Text Available Recently, there has been multi-agency promotion of entomophagy as an environmentally-friendly source of food for the ever increasing human population especially in the developing countries. However, food quality and safety concerns must first be addressed in this context. We addressed these concerns in the present study using the edible stink bug Encosternum delegorguei, which is widely consumed in southern Africa. We analysed for mycotoxins, and health beneficials including antioxidants, amino acids and essential fatty acids using liquid chromatography coupled to quadrupole time of flight mass spectrometry (LC-Qtof-MS and coupled gas chromatography (GC-MS. We also performed proximate analysis to determine nutritional components. We identified the human carcinogen mycotoxin (aflatoxin B1 at low levels in edible stink bugs that were stored in traditonally woven wooden dung smeared baskets and gunny bags previously used to store cereals. However, it was absent in insects stored in clean zip lock bags. On the other hand, we identified 10 fatty acids, of which 7 are considered essential fatty acids for human nutrition and health; 4 flavonoids and 12 amino acids of which two are considered the most limiting amino acids in cereal based diets. The edible stink bug also contained high crude protein and fats but was a poor source of minerals, except for phosphorus which was found in relatively high levels. Our results show that the edible stink bug is a nutrient- and antioxidant-rich source of food and health benefits for human consumption. As such, use of better handling and storage methods can help eliminate contamination of the edible stink bug with the carcinogen aflatoxin and ensure its safety as human food.

Aflatoxin Contamination Detected in Nutrient and Anti-Oxidant Rich Edible Stink Bug Stored in Recycled Grain Containers.

Science.gov (United States)

Musundire, Robert; Osuga, Isaac M; Cheseto, Xavier; Irungu, Janet; Torto, Baldwyn

2016-01-01

Recently, there has been multi-agency promotion of entomophagy as an environmentally-friendly source of food for the ever increasing human population especially in the developing countries. However, food quality and safety concerns must first be addressed in this context. We addressed these concerns in the present study using the edible stink bug Encosternum delegorguei, which is widely consumed in southern Africa. We analysed for mycotoxins, and health beneficials including antioxidants, amino acids and essential fatty acids using liquid chromatography coupled to quadrupole time of flight mass spectrometry (LC-Qtof-MS) and coupled gas chromatography (GC)-MS. We also performed proximate analysis to determine nutritional components. We identified the human carcinogen mycotoxin (aflatoxin B1) at low levels in edible stink bugs that were stored in traditonally woven wooden dung smeared baskets and gunny bags previously used to store cereals. However, it was absent in insects stored in clean zip lock bags. On the other hand, we identified 10 fatty acids, of which 7 are considered essential fatty acids for human nutrition and health; 4 flavonoids and 12 amino acids of which two are considered the most limiting amino acids in cereal based diets. The edible stink bug also contained high crude protein and fats but was a poor source of minerals, except for phosphorus which was found in relatively high levels. Our results show that the edible stink bug is a nutrient- and antioxidant-rich source of food and health benefits for human consumption. As such, use of better handling and storage methods can help eliminate contamination of the edible stink bug with the carcinogen aflatoxin and ensure its safety as human food.

Comparison of ingestion and topical application of insecticides against the common bed bug, Cimex lectularius (Hemiptera: Cimicidae)

Science.gov (United States)

Sierras, Angela; Schal, Coby

2016-01-01

Background The global prevalence of Cimex lectularius infestations has challenged current intervention efforts, as pyrethroid resistance has become ubiquitous, availability of labeled insecticides for bed bugs is limited, and non-chemical treatment options, such as heat, are often unaffordable. We evaluated representative insecticides toward the goal of developing a novel, ingestible liquid bait for hematophagous arthropods. Results LC50 values were estimated for adult males and first instar nymphs of an insecticide-susceptible strain for abamectin, clothianidin, fipronil and indoxacarb, after ingestion from an in vitro feeder. LD50 values were calculated based on the ingested blood volume. Ingested abamectin, clothianidin and fipronil caused rapid mortality in both life stages. Fipronil was ∼43-fold more effective by ingestion than by topical application. Indoxacarb and its bioactive metabolite decarbomethoxyllated JW062 (DCJW) were ineffective at causing bed bug mortality even at concentrations as high as 1000 ng mL−1 blood. Conclusions Fipronil, clothianidin and abamectin have potential for being incorporated into a liquid bait for bed bug control; indoxacarb and DCJW were not effective. Bed bugs are a good candidate for an ingestible liquid bait because systemic formulations generally require less active ingredient than residual sprays, they remain contained and more effectively target hematophagous arthropods. PMID:27766740

Novel Infectivity-Enhanced Oncolytic Adenovirus with a Capsid-Incorporated Dual-Imaging Moiety for Monitoring Virotherapy in Ovarian Cancer

Directory of Open Access Journals (Sweden)

Kristopher J. Kimball

2009-09-01

Full Text Available We sought to develop a cancer-targeted, infectivity-enhanced oncolytic adenovirus that embodies a capsid-labeling fusion for non-invasive dual-modality imaging of ovarian cancer virotherapy. A functional fusion protein composed of fluorescent and nuclear imaging tags was genetically incorporated into the capsid of an infectivity-enhanced conditionally replicative adenovirus. Incorporation of herpes simplex virus thymidine kinase (HSV-tk and monomeric red fluorescent protein 1 (mRFP1 into the viral capsid and its genomic stability were verified by molecular analyses. Replication and oncolysis were evaluated in ovarian cancer cells. Fusion functionality was confirmed by in vitro gamma camera and fluorescent microscopy imaging. Comparison of tk-mRFP virus to single-modality controls revealed similar replication efficiency and oncolytic potency. Molecular fusion did not abolish enzymatic activity of HSV-tk as the virus effectively phosphorylated thymidine both ex vivo and in vitro. In vitro fluorescence imaging demonstrated a strong correlation between the intensity of fluorescent signal and cytopathic effect in infected ovarian cancer cells, suggesting that fluorescence can be used to monitor viral replication. We have in vitro validated a new infectivity-enhanced oncolytic adenovirus with a dual-imaging modality-labeled capsid, optimized for ovarian cancer virotherapy. The new agent could provide incremental gains toward climbing the barriers for achieving conditionally replicated adenovirus efficacy in human trials.

Cross-serotype immunity induced by immunization with a conserved rhinovirus capsid protein.

Directory of Open Access Journals (Sweden)

Nicholas Glanville

Full Text Available Human rhinovirus (RV infections are the principle cause of common colds and precipitate asthma and COPD exacerbations. There is currently no RV vaccine, largely due to the existence of ∼150 strains. We aimed to define highly conserved areas of the RV proteome and test their usefulness as candidate antigens for a broadly cross-reactive vaccine, using a mouse infection model. Regions of the VP0 (VP4+VP2 capsid protein were identified as having high homology across RVs. Immunization with a recombinant VP0 combined with a Th1 promoting adjuvant induced systemic, antigen specific, cross-serotype, cellular and humoral immune responses. Similar cross-reactive responses were observed in the lungs of immunized mice after infection with heterologous RV strains. Immunization enhanced the generation of heterosubtypic neutralizing antibodies and lung memory T cells, and caused more rapid virus clearance. Conserved domains of the RV capsid therefore induce cross-reactive immune responses and represent candidates for a subunit RV vaccine.

Characterization of the invariable residue 51 mutations of human immunodeficiency virus type 1 capsid protein on in vitro CA assembly and infectivity

Directory of Open Access Journals (Sweden)

Höglund Stefan

2007-09-01

Full Text Available Abstract Background The mature HIV-1 conical core formation proceeds through highly regulated protease cleavage of the Gag precursor, which ultimately leads to substantial rearrangements of the capsid (CAp24 molecule involving both inter- and intra-molecular contacts of the CAp24 molecules. In this aspect, Asp51 which is located in the N-terminal domain of HIV-1 CAp24 plays an important role by forming a salt-bridge with the free imino terminus Pro1 following proteolytic cleavage and liberation of the CAp24 protein from the Pr55Gag precursor. Thus, previous substitution mutation of Asp51 to alanine (D51A has shown to be lethal and that this invariable residue was found essential for tube formation in vitro, virus replication and virus capsid formation. Results We extended the above investigation by introducing three different D51 substitution mutations (D51N, D51E, and D51Q into both prokaryotic and eukaryotic expression systems and studied their effects on in vitro capsid assembly and virus infectivity. Two substitution mutations (D51E and D51N had no substantial effect on in vitro capsid assembly, yet they impaired viral infectivity and particle production. In contrast, the D51Q mutant was defective both for in vitro capsid assembly and for virus replication in cell culture. Conclusion These results show that substitutions of D51 with glutamate, glutamine, or asparagine, three amino acid residues that are structurally related to aspartate, could partially rescue both in vitro capsid assembly and intra-cellular CAp24 production but not replication of the virus in cultured cells.

Formation of newly synthesized adeno-associated virus capsids in the cell nucleus.

Science.gov (United States)

Bell, Peter; Vandenberghe, Luk H; Wilson, James M

2014-06-01

Adeno-associated virus (AAV) particles inside the nucleus of a HEK 293 cell are shown by electron microscopy. Cells have been triple-transfected for vector production and were analyzed for capsid formation three days later. Newly assembled particle are visible as seemingly unstructured conglomerates or crystal-like arrays.

Structures of Adenovirus Incomplete Particles Clarify Capsid Architecture and Show Maturation Changes of Packaging Protein L1 52/55k.

Science.gov (United States)

Condezo, Gabriela N; Marabini, Roberto; Ayora, Silvia; Carazo, José M; Alba, Raúl; Chillón, Miguel; San Martín, Carmen

2015-09-01

Adenovirus is one of the most complex icosahedral, nonenveloped viruses. Even after its structure was solved at near-atomic resolution by both cryo-electron microscopy and X-ray crystallography, the location of minor coat proteins is still a subject of debate. The elaborated capsid architecture is the product of a correspondingly complex assembly process, about which many aspects remain unknown. Genome encapsidation involves the concerted action of five virus proteins, and proteolytic processing by the virus protease is needed to prime the virion for sequential uncoating. Protein L1 52/55k is required for packaging, and multiple cleavages by the maturation protease facilitate its release from the nascent virion. Light-density particles are routinely produced in adenovirus infections and are thought to represent assembly intermediates. Here, we present the molecular and structural characterization of two different types of human adenovirus light particles produced by a mutant with delayed packaging. We show that these particles lack core polypeptide V but do not lack the density corresponding to this protein in the X-ray structure, thereby adding support to the adenovirus cryo-electron microscopy model. The two types of light particles present different degrees of proteolytic processing. Their structures provide the first glimpse of the organization of L1 52/55k protein inside the capsid shell and of how this organization changes upon partial maturation. Immature, full-length L1 52/55k is poised beneath the vertices to engage the virus genome. Upon proteolytic processing, L1 52/55k disengages from the capsid shell, facilitating genome release during uncoating. Adenoviruses have been extensively characterized as experimental systems in molecular biology, as human pathogens, and as therapeutic vectors. However, a clear picture of many aspects of their basic biology is still lacking. Two of these aspects are the location of minor coat proteins in the capsid and the

Antibody Competition Reveals Surface Location of HPV L2 Minor Capsid Protein Residues 17–36

Directory of Open Access Journals (Sweden)

Stephanie M. Bywaters

2017-11-01

Full Text Available The currently available nonavalent human papillomavirus (HPV vaccine exploits the highly antigenic L1 major capsid protein to promote high-titer neutralizing antibodies, but is limited to the HPV types included in the vaccine since the responses are highly type-specific. The limited cross-protection offered by the L1 virus-like particle (VLP vaccine warrants further investigation into cross-protective L2 epitopes. The L2 proteins are yet to be fully characterized as to their precise placement in the virion. Adding to the difficulties in localizing L2, studies have suggested that L2 epitopes are not well exposed on the surface of the mature capsid prior to cellular engagement. Using a series of competition assays between previously mapped anti-L1 monoclonal antibodies (mAbs (H16.V5, H16.U4 and H16.7E and novel anti-L2 mAbs, we probed the capsid surface for the location of an L2 epitope (aa17–36. The previously characterized L1 epitopes together with our competition data is consistent with a proposed L2 epitope within the canyons of pentavalent capsomers.

Immune responses of a native and an invasive bird to Buggy Creek Virus (Togaviridae: Alphavirus and its arthropod vector, the swallow bug (Oeciacus vicarius.

Directory of Open Access Journals (Sweden)

Carol A Fassbinder-Orth

Full Text Available Invasive species often display different patterns of parasite burden and virulence compared to their native counterparts. These differences may be the result of variability in host-parasite co-evolutionary relationships, the occurrence of novel host-parasite encounters, or possibly innate differences in physiological responses to infection between invasive and native hosts. Here we examine the adaptive, humoral immune responses of a resistant, native bird and a susceptible, invasive bird to an arbovirus (Buggy Creek virus; Togaviridae: Alphavirus and its ectoparasitic arthropod vector (the swallow bug; Oeciacus vicarius. Swallow bugs parasitize the native, colonially nesting cliff swallow (Petrochelidon pyrrhonota and the introduced house sparrow (Passer domesticus that occupies nests in cliff swallow colonies. We measured levels of BCRV-specific and swallow bug-specific IgY levels before nesting (prior to swallow bug exposure and after nesting (after swallow bug exposure in house sparrows and cliff swallows in western Nebraska. Levels of BCRV-specific IgY increased significantly following nesting in the house sparrow but not in the cliff swallow. Additionally, house sparrows displayed consistently higher levels of swallow bug-specific antibodies both before and after nesting compared to cliff swallows. The higher levels of BCRV and swallow bug specific antibodies detected in house sparrows may be reflective of significant differences in both antiviral and anti-ectoparasite immune responses that exist between these two avian species. To our knowledge, this is the first study to compare the macro- and microparasite-specific immune responses of an invasive and a native avian host exposed to the same parasites.

Investigating Information Needs to Improve Cooperation Between Developers and Bug Reporters

NARCIS (Netherlands)

Breu, S.; Premraj, R.; Sillito, J.; Zimmermann, T.

2010-01-01

For many software projects, bug tracking systems play a central role in supporting collaboration between the developers and the users of the software. To better understand this collaboration and how tool support can be improved, we have quantitatively and qualitatively analysed the questions asked

Ecosystem-based incorporation of nectar-producing plants for stink bug parasitoids

Science.gov (United States)

Adult parasitoids of pest insects rely on floral resources for survival and reproduction but can be food-deprived in intensively managed agricultural systems lacking these resources. Stink bugs are serious pests of crops in southwest Georgia. Provisioning nectar-producing plants for parasitoids of s...

An Efficient Technique for Bayesian Modelling of Family Data Using the BUGS software

Directory of Open Access Journals (Sweden)

Harold T Bae

2014-11-01

Full Text Available Linear mixed models have become a popular tool to analyze continuous data from family-based designs by using random effects that model the correlation of subjects from the same family. However, mixed models for family data are challenging to implement with the BUGS (Bayesian inference Using Gibbs Sampling software because of the high-dimensional covariance matrix of the random effects. This paper describes an efficient parameterization that utilizes the singular value decomposition of the covariance matrix of random effects, includes the BUGS code for such implementation, and extends the parameterization to generalized linear mixed models. The implementation is evaluated using simulated data and an example from a large family-based study is presented with a comparison to other existing methods.

Molecular characterization of genome segments 1 and 3 encoding two capsid proteins of Antheraea mylitta cytoplasmic polyhedrosis virus

Directory of Open Access Journals (Sweden)

Chakrabarti Mrinmay

2010-08-01

Full Text Available Abstract Background Antheraea mylitta cytoplasmic polyhedrosis virus (AmCPV, a cypovirus of Reoviridae family, infects Indian non-mulberry silkworm, Antheraea mylitta, and contains 11 segmented double stranded RNA (S1-S11 in its genome. Some of its genome segments (S2 and S6-S11 have been previously characterized but genome segments encoding viral capsid have not been characterized. Results In this study genome segments 1 (S1 and 3 (S3 of AmCPV were converted to cDNA, cloned and sequenced. S1 consisted of 3852 nucleotides, with one long ORF of 3735 nucleotides and could encode a protein of 1245 amino acids with molecular mass of ~141 kDa. Similarly, S3 consisted of 3784 nucleotides having a long ORF of 3630 nucleotides and could encode a protein of 1210 amino acids with molecular mass of ~137 kDa. BLAST analysis showed 20-22% homology of S1 and S3 sequence with spike and capsid proteins, respectively, of other closely related cypoviruses like Bombyx mori CPV (BmCPV, Lymantria dispar CPV (LdCPV, and Dendrolimus punctatus CPV (DpCPV. The ORFs of S1 and S3 were expressed as 141 kDa and 137 kDa insoluble His-tagged fusion proteins, respectively, in Escherichia coli M15 cells via pQE-30 vector, purified through Ni-NTA chromatography and polyclonal antibodies were raised. Immunoblot analysis of purified polyhedra, virion particles and virus infected mid-gut cells with the raised anti-p137 and anti-p141 antibodies showed specific immunoreactive bands and suggest that S1 and S3 may code for viral structural proteins. Expression of S1 and S3 ORFs in insect cells via baculovirus recombinants showed to produce viral like particles (VLPs by transmission electron microscopy. Immunogold staining showed that S3 encoded proteins self assembled to form viral outer capsid and VLPs maintained their stability at different pH in presence of S1 encoded protein. Conclusion Our results of cloning, sequencing and functional analysis of AmCPV S1 and S3 indicate that S3

An elementary introduction to Bayesian computing using WinBUGS.

Science.gov (United States)

Fryback, D G; Stout, N K; Rosenberg, M A

2001-01-01

Bayesian statistics provides effective techniques for analyzing data and translating the results to inform decision making. This paper provides an elementary tutorial overview of the WinBUGS software for performing Bayesian statistical analysis. Background information on the computational methods used by the software is provided. Two examples drawn from the field of medical decision making are presented to illustrate the features and functionality of the software.

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

Countercurrent heat exchange and thermoregulation during blood-feeding in kissing bugs.

Science.gov (United States)

Lahondère, Chloé; Insausti, Teresita C; Paim, Rafaela Mm; Luan, Xiaojie; Belev, George; Pereira, Marcos H; Ianowski, Juan P; Lazzari, Claudio R

2017-11-21

Blood-sucking insects experience thermal stress at each feeding event on endothermic vertebrates. We used thermography to examine how kissing-bugs Rhodnius prolixus actively protect themselves from overheating. During feeding, these bugs sequester and dissipate the excess heat in their heads while maintaining an abdominal temperature close to ambient. We employed a functional-morphological approach, combining histology, µCT and X-ray-synchrotron imaging to shed light on the way these insects manage the flow of heat across their bodies. The close alignment of the circulatory and ingestion systems, as well as other morphological characteristics, support the existence of a countercurrent heat exchanger in the head of R. prolixus , which decreases the temperature of the ingested blood before it reaches the abdomen. This kind of system has never been described before in the head of an insect. For the first time, we show that countercurrent heat exchange is associated to thermoregulation during blood-feeding.

Status of Urban Bed Bug Infestations in Southern China: An Analysis of Pest Control Service Records in Shenzhen in 2012 and Dongguan in 2013.

Science.gov (United States)

Wang, Lei; Cai, Xuquan; Xu, Yijuan

2015-01-01

The recent resurgence of bed bugs (Cimex spp.) in many developed countries has drawn increasing attention worldwide. The status of urban bed bug infestations were investigated in Shenzhen and Dongguan, two major cities in southern Guangdong Province of southern China, based on pest control service records from two different companies (one during 2012 and another during 2013). The results showed that Shenzhen and Dongguan have a severe problem with bed bug infestations: the control of bed bugs is a constant concern, except during the winter. In Shenzhen, a similar number of premises were treated for bed bugs in central business districts and suburban districts. However, in Dongguan, more premises were treated for bed bugs in suburban districts than in central business districts. The treatment rate for worker sleeping quarters, apartments, hotel, and private houses in Shenzhen was 53.8, 43.0, 1.9, and 1.3%, respectively. The percentage of treated rooms was 56.1% for worker sleeping quarters and 91.1% for apartments. In Dongguan, the treatment rate for worker sleeping quarters, apartments, hotel, and private houses was 90.0, 10.0, 0.0, and 0.0%, respectively. © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Reactions of green lizards (Lacerta viridis) to major repellent compounds secreted by Graphosoma lineatum (Heteroptera: Pentatomidae).

Science.gov (United States)

Gregorovičová, Martina; Černíková, Alena

2015-06-01

The chemical defence of Heteroptera is primarily based on repellent secretions which signal the potential toxicity of the bug to its predators. We tested the aversive reactions of green lizards (Lacerta viridis) towards the major compounds of the defensive secretion of Graphosoma lineatum, specifically: (i) a mixture of three aldehydes: (E)-hex-2-enal, (E)-oct-2-enal, (E)-dec-2-enal; (ii) a mixture of these three aldehydes and tridecane; (iii) oxoaldehyde: (E)-4-oxohex-2-enal; (iv) secretion extracted from metathoracic scent glands of G. lineatum adults and (v) hexane as a non-polar solvent. All chemicals were presented on a palatable food (Tenebrio molitor larvae). The aversive reactions of the green lizards towards the mealworms were evaluated by observing the approach latencies, attack latencies and approach-attack intervals. The green lizards exhibited a strong aversive reaction to the mixture of three aldehydes. Tridecane reduced the aversive reaction to the aldehyde mixture. Oxoaldehyde caused the weakest, but still significant, aversive reaction. The secretion from whole metathoracic scent glands also clearly had an aversive effect on the green lizards. Moreover, when a living specimen of G. lineatum or Pyrrhocoris apterus (another aposematic red-and-black prey) was presented to the green lizards before the trials with the aldehyde mixture, the aversive effect of the mixture was enhanced. In conclusion, the mixture of three aldehydes had the strong aversive effect and could signal the potential toxicity of G. lineatum to the green lizards. Copyright © 2015 Elsevier GmbH. All rights reserved.

Influence of Type of Electric Bright Light on the Attraction of the African Giant Water Bug, Lethocerus indicus (Hemiptera: Belostomatidae

Directory of Open Access Journals (Sweden)

Luke Chinaru Nwosu

2012-01-01

Full Text Available This study investigated the influence of type of electric bright light (produced by fluorescent light tube and incandescent light bulb on the attraction of the African giant water bug, Lethocerus indicus (Hemiptera: Belostomatidae. Four fluorescent light tubes of 15 watts each, producing white-coloured light and four incandescent light bulbs of 60 watts each, producing yellow-coloured light, but both producing the same amount of light, were varied and used for the experiments. Collections of bugs at experimental house were done at night between the hours of 8.30 pm and 12 mid-night on daily basis for a period of four months per experiment in the years 2008 and 2009. Lethocerus indicus whose presence in any environment has certain implications was the predominant belostomatid bug in the area. Use of incandescent light bulbs in 2009 significantly attracted more Lethocerus indicus 103 (74.6% than use of fluorescent light tubes 35 (25.41% in 2008 [4.92=0.0001]. However, bug’s attraction to light source was not found sex dependent [>0.05; (>0.18=0.4286 and >0.28=0.3897]. Therefore, this study recommends the use of fluorescent light by households, campgrounds, and other recreational centres that are potentially exposed to the nuisance of the giant water bugs. Otherwise, incandescent light bulbs should be used when it is desired to attract the presence of these aquatic bugs either for food or scientific studies.

Detention of HPV L1 Capsid Protein and hTERC Gene in Screening of Cervical Cancer

Directory of Open Access Journals (Sweden)

Huang Bin

2013-06-01

Full Text Available   Objective(s: To investigate the expression of human papilloma virus (HPV L1 capsid protein, and human telomerase RNA component (hTERC in cervical cancer and the role of detection of both genes in screening of cervical cancer.   Materials and Methods: A total of 309 patients were recruited and cervical exfoliated cells were collected. Immunocytochemistry was employed to detect HPV L1 capsid protein, and fluorescent in situ hybridization (FISH was performed to detect the hTERC. Results: The expression of HPV L1 capsid protein reduced with the increase of the histological grade of cervical cells and was negatively related to the grade of cervical lesions. However, the expression of hTERC increased with the increase of the histological grade and positively associated with the grade of cervical lesions. The proportion of patients with L1(-/hTERC(+ was higher in patients with histological grade of CIN2 or higher than that in those with histological grade of CIN1. The L1(+/hTERC(- and L1(-/hTERC(- were negatively related to the grade of cervical lesions. L1(-/hTERC(+ was positively associated with the grade of cervical lesions. The L1/hTERC ratio increased. The negative predictive value of both HPV L1 and hTERC was higher than that of HPV L1 or hTERC, but there was no marked difference in the screening efficacy of cervical cancer among HPV L1, hTERC and HPV L1+hTERC. Conclusion: HPV L1 capsid protein and hTERC gene may serve as markers for the early diagnosis and prediction of cervical lesions. The increase in L1/hTERC ratio reflects the progression of cervical lesions to a certain extent.

Structures of foot and mouth disease virus pentamers: Insight into capsid dissociation and unexpected pentamer reassociation.

Directory of Open Access Journals (Sweden)

Nayab Malik

2017-09-01

Full Text Available Foot-and-mouth disease virus (FMDV belongs to the Aphthovirus genus of the Picornaviridae, a family of small, icosahedral, non-enveloped, single-stranded RNA viruses. It is a highly infectious pathogen and is one of the biggest hindrances to the international trade of animals and animal products. FMDV capsids (which are unstable below pH6.5 release their genome into the host cell from an acidic compartment, such as that of an endosome, and in the process dissociate into pentamers. Whilst other members of the family (enteroviruses have been visualized to form an expanded intermediate capsid with holes from which inner capsid proteins (VP4, N-termini (VP1 and RNA can be released, there has been no visualization of any such state for an aphthovirus, instead the capsid appears to simply dissociate into pentamers. Here we present the 8-Å resolution structure of isolated dissociated pentamers of FMDV, lacking VP4. We also found these pentamers to re-associate into a rigid, icosahedrally symmetric assembly, which enabled their structure to be solved at higher resolution (5.2 Å. In this assembly, the pentamers unexpectedly associate 'inside out', but still with their exposed hydrophobic edges buried. Stabilizing interactions occur between the HI loop of VP2 and its symmetry related partners at the icosahedral 3-fold axes, and between the BC and EF loops of VP3 with the VP2 βB-strand and the CD loop at the 2-fold axes. A relatively extensive but subtle structural rearrangement towards the periphery of the dissociated pentamer compared to that in the mature virus provides insight into the mechanism of dissociation of FMDV and the marked difference in antigenicity.

Verde plant bug (Hemiptera: Miridae) feeding injury to cotton bolls characterized by boll age, size, and damage ratings.

Science.gov (United States)

Armstrong, J Scott; Brewer, Michael J; Parker, Roy D; Adamczyk, J J

2013-02-01

The verde plant bug, Creontiades signatus (Distant), has been present in south Texas for several years but has more recently been documented as an economic threat to cultivated cotton, (Gossypium hirsutum L. Our studies over 2 yr (2009 and 2010) and two locations (Weslaco and Corpus Christi, TX) investigated feeding-injury of the verde plant bug to a range of cotton boll age classes defined by boll diameter and accumulated degree-days (anthesis to the time of infesting) for first-position cotton bolls infested with the plant bugs. The most detrimental damage to younger cotton holls from verde plant bug feeding was boll abscission. Cotton bolls verde plant bug injured bolls compared with the controls up to 162 ACDD or a mean boll diameter 2.0 cm. Cotton seed weights were significantly reduced up to 179 ACDD or a boll diameter of 2.0 cm at Weslaco in 2009, and up to 317 ACDD or boll diameter 2.6 cm for Weslaco in 2010 when compared with the noninfested controls. Lint weight per cotton boll for infested and noninfested bolls was significantly reduced up to 262 ACDD or boll diameter 2.5 for Corpus Christi in 2010 and up to 288 ACCD or boll diameter 2.6 cm for Weslaco, TX, in 2010. Damage ratings (dependant variable) regressed against infested and noninfested seed-cotton weights showed that in every instance, the infested cotton bolls had a strong and significant relationship with damage ratings for all age classes of bolls. Damage ratings for the infested cotton bolls that did not abscise by harvest showed visual signs of verde plant bug feeding injury and the subsequent development ofboll rot; however, these two forms of injury causing lint and seed mass loss are hard to differentiate from open or boll-locked cotton bolls. Based on the results of both lint and seed loss over 2 yr and four studies cotton bolls should be protected up to approximately 300 ACDD or a boll diameter of 2.5 cm. This equilibrates to bolls that are 12-14 d of age dependent upon daily maximum

The Heteroptera of the Netherlands Antilles – V Tingidae (Lace Bugs)

NARCIS (Netherlands)

Drake, Carl J.; Cobben, R.H.

1960-01-01

The present paper is based upon the lace bugs, Family Tingidae, collected by the junior author in the West Indies, on the islands of Aruba, Curaçao, Bonaire, St. Martin, Saba, and St. Eustatius. This collection of several tingids comprises 17 species, including the five new forms described below.

Metabolic Resistance in Bed Bugs

Directory of Open Access Journals (Sweden)

Omprakash Mittapalli

2011-03-01

Full Text Available Blood-feeding insects have evolved resistance to various insecticides (organochlorines, pyrethroids, carbamates, etc. through gene mutations and increased metabolism. Bed bugs (Cimex lectularius are hematophagous ectoparasites that are poised to become one of the major pests in households throughout the United States. Currently, C. lectularius has attained a high global impact status due to its sudden and rampant resurgence. Resistance to pesticides is one factor implicated in this phenomenon. Although much emphasis has been placed on target sensitivity, little to no knowledge is available on the role of key metabolic players (e.g., cytochrome P450s and glutathione S-transferases towards pesticide resistance in C. lectularius. In this review, we discuss different modes of resistance (target sensitivity, penetration resistance, behavioral resistance, and metabolic resistance with more emphasis on metabolic resistance.

Towards green loyalty: the influences of green perceived risk, green image, green trust and green satisfaction

Science.gov (United States)

Chrisjatmiko, K.

2018-01-01

The paper aims to present a comprehensive framework for the influences of green perceived risk, green image, green trust and green satisfaction to green loyalty. The paper also seeks to account explicitly for the differences in green perceived risk, green image, green trust, green satisfaction and green loyalty found among green products customers. Data were obtained from 155 green products customers. Structural equation modeling was used in order to test the proposed hypotheses. The findings show that green image, green trust and green satisfaction has positive effects to green loyalty. But green perceived risk has negative effects to green image, green trust and green satisfaction. However, green perceived risk, green image, green trust and green satisfaction also seems to be a good device to gain green products customers from competitors. The contributions of the paper are, firstly, a more complete framework of the influences of green perceived risk, green image, green trust and green satisfaction to green loyalty analyses simultaneously. Secondly, the study allows a direct comparison of the difference in green perceived risk, green image, green trust, green satisfaction and green loyalty between green products customers.

Utilization of early soybeans for food and reproduction by the tarnished plant bug (Hemiptera: Miridae) in the delta of Mississippi.

Science.gov (United States)

Snodgrass, G L; Jackson, R E; Abel, C A; Perera, O P

2010-08-01

Commercially produced maturity group (MG) IV soybeans, Glycine max L., were sampled during bloom for tarnished plant bugs, Lygus lineolaris (Palisot de Beauvois), during May and June 1999 (3 fields) and 2001 (18 fields). The adults and nymphs were found primarily in single population peaks in both years, indicating a single new generation was produced during each year. The peak mean numbers of nymphs were 0.61 and 0.84 per drop cloth sample in 1999 and 2001, respectively. Adults peaked at 3.96 (1999) and 3.76 (2001) per sweep net sample (25 sweeps). Tests using laboratory-reared and field-collected tarnished plant bugs resulted in very poor survival of nymphs on 16 different soybean varieties (MG III, one; IV, four; V, nine; VI, two). A large cage (0.06 ha) field test found that the number of nymphs produced on eight soybean varieties after mated adults were released into the cages was lower than could be expected on a suitable host. These results indicated that soybean was a marginal host for tarnished plant bugs. However, the numbers of adults and nymphs found in the commercially produced fields sampled in the study may have been high enough to cause feeding damage to the flowering soybeans. The nature of the damage and its possible economic importance were not determined. Reproduction of tarnished plant bugs in the commercially produced early soybean fields showed that the early soybeans provided tarnished plant bugs with a very abundant host at a time when only wild hosts were previously available.

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

76 FR 550 - Second National Bed Bug Summit; Notice of Public Meeting

Science.gov (United States)

2011-01-05

... identify knowledge gaps and barriers to effective community-wide bed bug control; propose the next steps in addressing knowledge gaps and eliminating barriers; and develop a framework for addressing the highest... many areas of the country. As this resilient pest has become a nationwide problem affecting hotels...

Specific interaction between hnRNP H and HPV16 L1 proteins: Implications for late gene auto-regulation enabling rapid viral capsid protein production

Energy Technology Data Exchange (ETDEWEB)

Zheng, Zi-Zheng; Sun, Yuan-Yuan; Zhao, Min; Huang, Hui [National Institute of Diagnostics and Vaccine Development in Infectious Diseases, Xiamen University, Xiamen, Fujian 361005 (China); School of Life Sciences, Xiamen University, Xiamen, Fujian 361005 (China); Zhang, Jun; Xia, Ning-Shao [National Institute of Diagnostics and Vaccine Development in Infectious Diseases, Xiamen University, Xiamen, Fujian 361005 (China); School of Life Sciences, Xiamen University, Xiamen, Fujian 361005 (China); School of Public Health, Xiamen University, Xiamen, Fujian 361005 (China); Miao, Ji, E-mail: jmiao@xmu.edu.cn [National Institute of Diagnostics and Vaccine Development in Infectious Diseases, Xiamen University, Xiamen, Fujian 361005 (China); School of Life Sciences, Xiamen University, Xiamen, Fujian 361005 (China); Zhao, Qinjian, E-mail: qinjian_zhao@xmu.edu.cn [National Institute of Diagnostics and Vaccine Development in Infectious Diseases, Xiamen University, Xiamen, Fujian 361005 (China); School of Public Health, Xiamen University, Xiamen, Fujian 361005 (China)

2013-01-18

Highlights: ► The RNA-binding hnRNP H regulates late viral gene expression. ► hnRNP H activity was inhibited by a late viral protein. ► Specific interaction between HPV L1 and hnRNP H was demonstrated. ► Co-localization of HPV L1 and hnRNP H inside cells was observed. ► Viral capsid protein production, enabling rapid capsid assembly, was implicated. -- Abstract: Heterogeneous nuclear ribonucleoproteins (hnRNPs), including hnRNP H, are RNA-binding proteins that function as splicing factors and are involved in downstream gene regulation. hnRNP H, which binds to G triplet regions in RNA, has been shown to play an important role in regulating the staged expression of late proteins in viral systems. Here, we report that the specific association between hnRNP H and a late viral capsid protein, human papillomavirus (HPV) L1 protein, leads to the suppressed function of hnRNP H in the presence of the L1 protein. The direct interaction between the L1 protein and hnRNP H was demonstrated by complex formation in solution and intracellularly using a variety of biochemical and immunochemical methods, including peptide mapping, specific co-immunoprecipitation and confocal fluorescence microscopy. These results support a working hypothesis that a late viral protein HPV16 L1, which is down regulated by hnRNP H early in the viral life cycle may provide an auto-regulatory positive feedback loop that allows the rapid production of HPV capsid proteins through suppression of the function of hnRNP H at the late stage of the viral life cycle. In this positive feedback loop, the late viral gene products that were down regulated earlier themselves disable their suppressors, and this feedback mechanism could facilitate the rapid production of capsid proteins, allowing staged and efficient viral capsid assembly.

Dengue Virus Capsid Protein Binds Core Histones and Inhibits Nucleosome Formation in Human Liver Cells

Science.gov (United States)

Colpitts, Tonya M.; Barthel, Sebastian; Wang, Penghua; Fikrig, Erol

2011-01-01

Dengue virus (DENV) is a member of the Flaviviridae and a globally (re)emerging pathogen that causes serious human disease. There is no specific antiviral or vaccine for dengue virus infection. Flavivirus capsid (C) is a structural protein responsible for gathering the viral RNA into a nucleocapsid that forms the core of a mature virus particle. Flaviviral replication is known to occur in the cytoplasm yet a large portion of capsid protein localizes to the nucleus during infection. The reasons for the nuclear presences of capsid are not completely understood. Here, we expressed mature DENV C in a tandem affinity purification assay to identify potential binding partners in human liver cells. DENV C targeted the four core histones, H2A, H2B, H3 and H4. DENV C bound recombinant histones in solution and colocalized with histones in the nucleus and cytoplasm of liver cells during DENV infection. We show that DENV C acts as a histone mimic, forming heterodimers with core histones, binding DNA and disrupting nucleosome formation. We also demonstrate that DENV infection increases the amounts of core histones in livers cells, which may be a cellular response to C binding away the histone proteins. Infection with DENV additionally alters levels of H2A phosphorylation in a time-dependent manner. The interactions of C and histones add an interesting new role for the presence of C in the nucleus during DENV infection. PMID:21909430

Breaking Symmetry in Viral Icosahedral Capsids as Seen through the Lenses of X-ray Crystallography and Cryo-Electron Microscopy

Directory of Open Access Journals (Sweden)

Kristin N. Parent

2018-02-01

Full Text Available The majority of viruses on Earth form capsids built by multiple copies of one or more types of a coat protein arranged with 532 symmetry, generating an icosahedral shell. This highly repetitive structure is ideal to closely pack identical protein subunits and to enclose the nucleic acid genomes. However, the icosahedral capsid is not merely a passive cage but undergoes dynamic events to promote packaging, maturation and the transfer of the viral genome into the host. These essential processes are often mediated by proteinaceous complexes that interrupt the shell’s icosahedral symmetry, providing a gateway through the capsid. In this review, we take an inventory of molecular structures observed either internally, or at the 5-fold vertices of icosahedral DNA viruses that infect bacteria, archea and eukaryotes. Taking advantage of the recent revolution in cryo-electron microscopy (cryo-EM and building upon a wealth of crystallographic structures of individual components, we review the design principles of non-icosahedral structural components that interrupt icosahedral symmetry and discuss how these macromolecules play vital roles in genome packaging, ejection and host receptor-binding.

Engineering bacterial surface displayed human norovirus capsid proteins: A novel system to explore interaction between norovirus and ligands

Directory of Open Access Journals (Sweden)

Mengya eNiu

2015-12-01

Full Text Available Human noroviruses (HuNoVs are major contributors to acute nonbacterial gastroenteritis outbreaks. Many aspects of HuNoVs are poorly understood due to both the current inability to culture HuNoVs, and the lack of efficient small animal models. Surrogates for HuNoVs, such as recombinant viral like particles (VLPs expressed in eukaryotic system or P particles expressed in prokaryotic system, have been used for studies in immunology and interaction between the virus and its receptors. However, it is difficult to use VLPs or P particles to collect or isolate potential ligands binding to these recombinant capsid proteins. In this study, a new strategy was used to collect HuNoVs binding ligands through the use of ice nucleation protein (INP to display recombinant capsid proteins of HuNoVs on bacterial surfaces. The viral protein-ligand complex could be easily separated by a low speed centrifugation step. This system was also used to explore interaction between recombinant capsid proteins of HuNoVs and their receptors. In this system, the VP1 capsid encoding gene (ORF2 and the protruding domain (P domain encoding gene (3’ terminal fragment of ORF2 of HuNoVs GI.1 and GII.4 were fused with 5’ terminal fragment of ice nucleation protein encoding gene (inaQn. The results demonstrated that the recombinant VP1 and P domains of HuNoVs were expressed and anchored on the surface of Escherichia coli BL21 cells after the bacteria were transformed with the corresponding plasmids. Both cell surface displayed VP1 and P domains could be recognized by HuNoVs specific antibodies and interact with the viral histo-blood group antigens receptors. In both cases, displayed P domains had better binding abilities than VP1. This new strategy of using displayed HuNoVs capsid proteins on the bacterial surface could be utilized to separate HuNoVs binding components from complex samples, to investigate interaction between the virus and its receptors, as well as to develop an

Structural determination of importin alpha in complex with beak and feather disease virus capsid nuclear localization signal

International Nuclear Information System (INIS)

Patterson, Edward I.; Dombrovski, Andrew K.; Swarbrick, Crystall M.D.; Raidal, Shane R.; Forwood, Jade K.

2013-01-01

Highlights: •Circovirus capsid proteins contain large nuclear localization signals (NLS). •A method of nuclear import has not been elucidated. •Beak and feather disease virus (BFDV) capsid NLS was crystallized with importin α. •The structure showed BFDV NLS binding to the major site of importin α. •Result shows implications for mechanism of nuclear transport for all circoviruses. -- Abstract: Circoviruses represent a rapidly increasing genus of viruses that infect a variety of vertebrates. Replication requires shuttling viral molecules into the host cell nucleus, a process facilitated by capsid-associated protein (Cap). Whilst a nuclear localization signal (NLS) has been shown to mediate nuclear translocation, the mode of nuclear transport remains to be elucidated. To better understand this process, beak and feather disease virus (BFDV) Cap NLS was crystallized with nuclear import receptor importin-α (Impα). Diffraction yielded structural data to 2.9 Å resolution, and the binding site on both Impα and BFDV Cap NLS were well resolved. The binding mechanism for the major site is likely conserved across circoviruses as supported by the similarity of NLSs in circovirus Caps. This finding illuminates a crucial step for infection of host cells by this viral family, and provides a platform for rational drug design against the binding interface

Evaluating impacts of brown marmorated stink bug on non-fruiting nursery crops

Science.gov (United States)

Halyomorpha halys, commonly known as the brown marmorated stink bug (BMSB), has become a major pest and nuisance since it arrived in the US in 1998 for both agricultural growers and homeowners. They can feed on ~200 different plant species, several of which are important ornamental crop species. The...

Promoter analysis of the Chilo iridescent virus DNA polymerase and major capsid protein genes

International Nuclear Information System (INIS)

Nalcacioglu, Remziye; Marks, Hendrik; Vlak, Just M.; Demirbag, Zihni; Oers, Monique M. van

2003-01-01

The DNA polymerase (DNApol) and major capsid protein (MCP) genes were used as models to study promoter activity in Chilo iridescent virus (CIV). Infection of Bombyx mori SPC-BM-36 cells in the presence of inhibitors of DNA or protein synthesis showed that DNApol, as well as helicase, is an immediate-early gene and confirmed that the major capsid protein (MCP) is a late gene. Transcription of DNApol initiated 35 nt upstream and that of MCP 14 nt upstream of the translational start site. In a luciferase reporter gene assay both promoters were active only when cells were infected with CIV. For DNApol sequences between position -27 and -6, relative to the transcriptional start site, were essential for promoter activity. Furthermore, mutation of a G within the sequence TTGTTTT located just upstream of the DNApol transcription initiation site reduced the promoter activity by 25%. Sequences crucial for MCP promoter activity are located between positions -53 and -29

Populations of Rice Grain Bug, Paraeuscosmetus pallicomis, (Hemiptera: Lygaeidae) in Weed-free Paddy Field, Weedy Paddy Field and Paddy Dykes.

Science.gov (United States)

Abdullah, Tamrin; Nasruddin, Andi; Agus, Nurariaty

2017-07-01

Research on the populations of rice grain bug Paraeuscosmetus pallicomis Dallas (Hemiptera: Lygaeidae) in paddy field ecosystems was performed with the aim to determine the populations of rice grain bug in weed-free paddy field, weedy paddy field, and paddy dykes. Experiment was carried out in the village of Paccellekang in the district of Patallasang of Gowa Regency in South Sulawesi, Indonesia. Observations were performed during the milky grain stage (85 days after planting), the mature grain stage (105 days after planting), and one day after harvest (115 days after transplanting). Results showed that 85 days after the transplanting, the populations of rice grain bug was significantly higher in the weedy paddy field compared to weed-free field and paddy dykes with total numbers of 1.75, 3.53, and 0.31 insects per 2 hills, respectively. Similarly, 105 days after the transplanting, 2.53, 5.53, and 0.11 insects per hill, respectively. However, one day after the harvest (115 days after transplanting) the number of insects in weed-free field decreased, while in the dykes increased, and the weedy plot still had the highest number of insects per 2 hills. Our results suggested that weeds played an important role in regulating the bug population by providing alternative shelter and foods for the insect.

Mapping the Structural Determinants Responsible for Enhanced T Cell Activation to the Immunogenic Adeno-Associated Virus Capsid from Isolate Rhesus 32.33

Science.gov (United States)

Mays, Lauren E.; Wang, Lili; Tenney, Rebeca; Bell, Peter; Nam, Hyun-Joo; Lin, Jianping; Gurda, Brittney; Van Vliet, Kim; Mikals, Kyle; Agbandje-McKenna, Mavis

2013-01-01

Avoiding activation of immunity to vector-encoded proteins is critical to the safe and effective use of adeno-associated viral (AAV) vectors for gene therapy. While commonly used serotypes, such as AAV serotypes 1, 2, 7, 8, and 9, are often associated with minimal and/or dysfunctional CD8+ T cell responses in mice, the threshold for immune activation appears to be lower in higher-order species. We have modeled this discrepancy within the mouse by identifying two capsid variants with differential immune activation profiles: AAV serotype 8 (AAV8) and a hybrid between natural rhesus isolates AAVrh32 and AAVrh33 (AAVrh32.33). Here, we aimed to characterize the structural determinants of the AAVrh32.33 capsid that augment cellular immunity to vector-encoded proteins or those of AAV8 that may induce tolerance. We hypothesized that the structural domain responsible for differential immune activation could be mapped to surface-exposed regions of the capsid, such as hypervariable regions (HVRs) I to IX of VP3. To test this, a series of hybrid AAV capsids was constructed by swapping domains between AAV8 and AAVrh32.33. By comparing their ability to generate transgene-specific T cells in vivo versus the stability of transgene expression in the muscle, we confirmed that the functional domain lies within the VP3 portion of the capsid. Our studies were able to exclude the regions of VP3 which are not sufficient for augmenting the cellular immune response, notably, HVRs I, II, and V. We have also identified HVR IV as a region of interest in conferring the efficiency and stability of muscle transduction to AAVrh32.33. PMID:23720715

Production of highly knotted DNA by means of cosmid circularization inside phage capsids

Directory of Open Access Journals (Sweden)

Trigueros Sonia

2007-12-01

Full Text Available Abstract Background The formation of DNA knots is common during biological transactions. Yet, functional implications of knotted DNA are not fully understood. Moreover, potential applications of DNA molecules condensed by means of knotting remain to be explored. A convenient method to produce abundant highly knotted DNA would be highly valuable for these studies. Results We had previously shown that circularization of the 11.2 kb linear DNA of phage P4 inside its viral capsid generates complex knots by the effect of confinement. We demonstrate here that this mechanism is not restricted to the viral genome. We constructed DNA cosmids as small as 5 kb and introduced them inside P4 capsids. Such cosmids were then recovered as a complex mixture of highly knotted DNA circles. Over 250 μg of knotted cosmid were typically obtained from 1 liter of bacterial culture. Conclusion With this biological system, DNA molecules of varying length and sequence can be shaped into very complex and heterogeneous knotted forms. These molecules can be produced in preparative amounts suitable for systematic studies and applications.

Genetically Thermo-Stabilised, Immunogenic Poliovirus Empty Capsids; a Strategy for Non-replicating Vaccines.

Directory of Open Access Journals (Sweden)

Helen Fox

2017-01-01

Full Text Available While wild type polio has been nearly eradicated there will be a need to continue immunisation programmes for some time because of the possibility of re-emergence and the existence of long term excreters of poliovirus. All vaccines in current use depend on growth of virus and most of the non-replicating (inactivated vaccines involve wild type viruses known to cause poliomyelitis. The attenuated vaccine strains involved in the eradication programme have been used to develop new inactivated vaccines as production is thought safer. However it is known that the Sabin vaccine strains are genetically unstable and can revert to a virulent transmissible form. A possible solution to the need for virus growth would be to generate empty viral capsids by recombinant technology, but hitherto such particles are so unstable as to be unusable. We report here the genetic manipulation of the virus to generate stable empty capsids for all three serotypes. The particles are shown to be extremely stable and to generate high levels of protective antibodies in animal models.

The complex subcellular distribution of satellite panicum mosaic virus capsid protein reflects its multifunctional role during infection

International Nuclear Information System (INIS)

Qi Dong; Omarov, Rustem T.; Scholthof, Karen-Beth G.

2008-01-01

Satellite panicum mosaic virus (SPMV) depends on its helper Panicum mosaic virus for replication and movement in host plants. The positive-sense single-stranded genomic RNA of SPMV encodes a 17-kDa capsid protein (CP) to form 16-nm virions. We determined that SPMV CP accumulates in both cytosolic and non-cytosolic fractions, but cytosolic accumulation of SPMV CP is exclusively associated with virions. An N-terminal arginine-rich motif (N-ARM) on SPMV CP is used to bind its cognate RNA and to form virus particles. Intriguingly, virion formation is dispensable for successful systemic SPMV RNA accumulation, yet this process still depends on an intact N-ARM. In addition, a C-terminal domain on the SPMV CP is necessary for self-interaction. Biochemical fractionation and fluorescent microscopy of green fluorescent protein-tagged SPMV CP demonstrated that the non-cytosolic SPMV CP is associated with the cell wall, the nucleus and other membranous organelles. To our knowledge, this is the first report that a satellite virus CP not only accumulates exclusively as virions in the cytosol but also is directed to the nucleolus and membranes. That SPMV CP is found both in the nucleus and the cell wall suggests its involvement in viral nuclear import and cell-to-cell transport

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...

Scale insects and mealy bugs (Homoptera: Coccoidea) attacking deciduous fruit trees in the western north coast of Alexandria, Egypt.

Science.gov (United States)

Mourad, A K; Moursi Khadiga, S; Mesbah, H A; Abdel-Razak Soad, I

2008-01-01

This investigation covered a survey of scale insects and mealy bugs infesting ten growing species of deciduous fruit trees in three localities in Alexandria govemorate. These localities were Merghem, Burg El-Arab, and El-Nahda about 50 Km. West of Alexandria under both rain-fed and irrigation system conditions. The common inspected fruit trees were fig, white mulberry, pomegranate, apple, pear, apricot, European plum, peach, almond, and persimmon. It was shown that a group of twenty scale insects and meaty bug species pertaining to fifteen genera belonging to six families of the super family: Coccoidea were collected and identified during the elapsing period from January to December, 2004. Among these species, Diaspidiotus perniciosus (Comstock) was recorded for the first time in Egypt. In the present study, many insect and non-insect parasitoids and predators were also found associated with these scale insects and mealy bugs on deciduous fruit trees in the three concerned localities throughout this investigation. These natural enemies were identified and recorded.

Toxicity of Bifenthrin and Mixtures of Bifenthrin Plus Acephate, Imidacloprid, Thiamethoxam, or Dicrotophos to Adults of Tarnished Plant Bug (Hemiptera: Miridae).

Science.gov (United States)

Jones, Moneen M; Duckworth, Jessica L; Robertson, Jacqueline

2018-04-02

To assess the toxicity of bifenthrin and four mixtures of insecticides to tarnished plant bug, we used an insecticide dip method of green bean to treat adults of a laboratory colony; mortality was assessed after 48 h. LC50s for imidacloprid, bifenthrin, acephate, thiamethoxam, and dicrotophos were 0.12, 0.39, 0.62, 0.67, and 3.96 ppm, respectively. LC75s for imidacloprid, bifenthrin, acephate, thiamethoxam, and dicrotophos were 0.61, 4.22, 5.10, 2.65, and 7.86 ppm, respectively. Based on the LC50s and LC75s, dicrotophos was much less toxic than the other chemicals tested. PoloMix software was used to determine syngerism, antagonism, or addition effects of the mixtures. Three out of four analyses of the joint action of bifenthrin plus imidacloprid or acephate or dicrotophos showed that toxicity was not independent and not correlated. For bifenthrin plus dicrotophos, observed mortality was greater than expected mortality at most concentrations suggesting synergism. Mixtures of bifenthrin plus imidacloprid and bifenthrin plus acephate showed observed mortality significantly less than expected, suggesting antagonism. LC50s for bifenthrin plus dicrotophos, acephate, imidacloprid, and thiamethoxam were 0.38, 1.06, 0.17, and 0.26 ppm, respectively. LC75s for bifenthrin plus dicrotophos, acephate, imidacloprid, and thiamethoxam were 13.61, 13.18, 0.67, and 0.80 ppm, respectively. Based on the LC50s and LC75s, bifenthrin plus acephate was 3- to 10-fold less toxic than the other chemicals tested. Bifenthrin plus acephate is frequently used in tank mixes to control tarnished plant bug and other cotton pests, and the effectiveness of each individual chemical appears to be reduced in one to one ratio mixtures.

Plant growth stage-specific injury and economic injury level for verde plant bug, Creontiades signatus (Hemiptera: Miridae), on cotton: effect of bloom period of infestation.

Science.gov (United States)

Brewer, Michael J; Anderson, Darwin J; Armstrong, J Scott

2013-10-01

Verde plant bugs, Creontiades signatus Distant (Hemiptera: Miridae), were released onto caged cotton, Cossypium hirsutum L., for a 1-wk period to characterize the effects of insect density and bloom period of infestation on cotton injury and yield in 2011 and 2012, Corpus Christi, TX. When plants were infested during early bloom (10-11 nodes above first white flower), a linear decline in fruit retention and boll load and a linear increase in boll injury were detected as verde plant bug infestation levels increased from an average of 0.5 to 4 bugs per plant. Lint and seed yield per plant showed a corresponding decline. Fruit retention, boll load, and yield were not affected on plants infested 1 wk later at peak bloom (8-9 nodes above first white flower), even though boll injury increased as infestation levels increased. Second-year testing verified boll injury but not yield loss, when infestations occurred at peak bloom. Incidence of cotton boll rot, known to be associated with verde plant bug feeding, was low to modest (verde plant bug were important contributors to yield decline, damage potential was greatest during the early bloom period of infestation, and a simple linear response best described the yield response-insect density relationship at early bloom. Confirmation that cotton after peak bloom was less prone to verde plant bug injury and an early bloom-specific economic injury level were key findings that can improve integrated pest management decision-making for dryland cotton, at least under low-rainfall growing conditions.

Promoter analysis of the Chilo iridescent virus DNA polymerase and major capsid protein genes

NARCIS (Netherlands)

Nalcacioglu, R.; Marks, H.; Vlak, J.M.; Demirbag, Z.; Oers, van M.M.

2003-01-01

The DNA polymerase (DNApol) and major capsid protein (MCP) genes were used as models to study promoter activity in Chilo iridescent virus (CIV). Infection of Bombyx mori SPC-BM-36 cells in the presence of inhibitors of DNA or protein synthesis showed that DNApol, as well as helicase, is an

The HACMS program: using formal methods to eliminate exploitable bugs.

Science.gov (United States)

Fisher, Kathleen; Launchbury, John; Richards, Raymond

2017-10-13

For decades, formal methods have offered the promise of verified software that does not have exploitable bugs. Until recently, however, it has not been possible to verify software of sufficient complexity to be useful. Recently, that situation has changed. SeL4 is an open-source operating system microkernel efficient enough to be used in a wide range of practical applications. Its designers proved it to be fully functionally correct, ensuring the absence of buffer overflows, null pointer exceptions, use-after-free errors, etc., and guaranteeing integrity and confidentiality. The CompCert Verifying C Compiler maps source C programs to provably equivalent assembly language, ensuring the absence of exploitable bugs in the compiler. A number of factors have enabled this revolution, including faster processors, increased automation, more extensive infrastructure, specialized logics and the decision to co-develop code and correctness proofs rather than verify existing artefacts. In this paper, we explore the promise and limitations of current formal-methods techniques. We discuss these issues in the context of DARPA's HACMS program, which had as its goal the creation of high-assurance software for vehicles, including quadcopters, helicopters and automobiles.This article is part of the themed issue 'Verified trustworthy software systems'. © 2017 The Authors.

The HACMS program: using formal methods to eliminate exploitable bugs

Science.gov (United States)

Launchbury, John; Richards, Raymond

2017-01-01

For decades, formal methods have offered the promise of verified software that does not have exploitable bugs. Until recently, however, it has not been possible to verify software of sufficient complexity to be useful. Recently, that situation has changed. SeL4 is an open-source operating system microkernel efficient enough to be used in a wide range of practical applications. Its designers proved it to be fully functionally correct, ensuring the absence of buffer overflows, null pointer exceptions, use-after-free errors, etc., and guaranteeing integrity and confidentiality. The CompCert Verifying C Compiler maps source C programs to provably equivalent assembly language, ensuring the absence of exploitable bugs in the compiler. A number of factors have enabled this revolution, including faster processors, increased automation, more extensive infrastructure, specialized logics and the decision to co-develop code and correctness proofs rather than verify existing artefacts. In this paper, we explore the promise and limitations of current formal-methods techniques. We discuss these issues in the context of DARPA’s HACMS program, which had as its goal the creation of high-assurance software for vehicles, including quadcopters, helicopters and automobiles. This article is part of the themed issue ‘Verified trustworthy software systems’. PMID:28871050

Natural insecticide based-leaves extract of Piper aduncum (Piperaceae in the control of stink bug brown soybean

Directory of Open Access Journals (Sweden)

Ludmila Porto Piton

2014-11-01

Full Text Available This research aimed to evaluate the insecticide potential of the leaves extract of Piper aduncum (L. (Piperaceae on development of brown stink bug, Euschistus heros (F., major pest in soybean crop in Mato Grosso State. For this a bioassay was conducted with eggs, nymphs and adults of this stink bug, using the concentrations 0.5; 1.0; 2.0; 4.0 and 8.0%, of the extract acetonic of P. aduncum and acetone for control. It was evaluated the numbers of unviable eggs, nymphs death and longevity, fecundity and fertility of the adults. In the immersing eggs bioassay, all concentrations greater than 1% unviable more than 70% of the eggs, while in the contact bioassay only 19% of the eggs were unviable in 8% concentration, differing of control. In nymphs bioassay the concentrations of 8% and 4% caused death of 72% and 52%, respectively. In adults bioassay all concentrations tested reduced significantly the survival and reproduction of the insects. Thus is evidence that P. aduncum is a potential plant for stink bug control, because it had deleterious action in all development phases on E. heros.

Controlled immobilisation of active enzymes on the cowpea mosaic virus capsid

Science.gov (United States)

Aljabali, Alaa A. A.; Barclay, J. Elaine; Steinmetz, Nicole F.; Lomonossoff, George P.; Evans, David J.

2012-08-01

Immobilisation of horseradish peroxidase (HRP) and glucose oxidase (GOX) via covalent attachment of modified enzyme carbohydrate to the exterior of the cowpea mosaic virus (CPMV) capsid gave high retention of enzymatic activity. The number of enzymes bound per virus was determined to be about eleven for HRP and 2-3 for GOX. This illustrates that relatively large biomacromolecules can be readily coupled to the virus surface using simple conjugation strategies. Virus-biomacromolecule hybrids have great potential for uses in catalysis, diagnostic assays or biosensors.Immobilisation of horseradish peroxidase (HRP) and glucose oxidase (GOX) via covalent attachment of modified enzyme carbohydrate to the exterior of the cowpea mosaic virus (CPMV) capsid gave high retention of enzymatic activity. The number of enzymes bound per virus was determined to be about eleven for HRP and 2-3 for GOX. This illustrates that relatively large biomacromolecules can be readily coupled to the virus surface using simple conjugation strategies. Virus-biomacromolecule hybrids have great potential for uses in catalysis, diagnostic assays or biosensors. Electronic supplementary information (ESI) available: Alternative conjugation strategies, agarose gel electrophoresis of CPMV and CPMV-HRP conjugates, UV-vis spectrum of HRP-ADHCPMV, agarose gel electrophoresis of GOX-ADHCPMV particles and corresponding TEM image, calibration curves for HRP-ADHCPMV and GOX-ADHCPMV, DLS data for GOX-ADHCPMV are made available. See DOI: 10.1039/c2nr31485a

Cryo-electron Microscopy Reconstruction and Stability Studies of the Wild Type and the R432A Variant of Adeno-associated Virus Type 2 Reveal that Capsid Structural Stability Is a Major Factor in Genome Packaging.

Science.gov (United States)

Drouin, Lauren M; Lins, Bridget; Janssen, Maria; Bennett, Antonette; Chipman, Paul; McKenna, Robert; Chen, Weijun; Muzyczka, Nicholas; Cardone, Giovanni; Baker, Timothy S; Agbandje-McKenna, Mavis

2016-10-01

The adeno-associated viruses (AAV) are promising therapeutic gene delivery vectors and better understanding of their capsid assembly and genome packaging mechanism is needed for improved vector production. Empty AAV capsids assemble in the nucleus prior to genome packaging by virally encoded Rep proteins. To elucidate the capsid determinants of this process, structural differences between wild-type (wt) AAV2 and a packaging deficient variant, AAV2-R432A, were examined using cryo-electron microscopy and three-dimensional image reconstruction both at an ∼5.0-Å resolution (medium) and also at 3.8- and 3.7-Å resolutions (high), respectively. The high resolution structures showed that removal of the arginine side chain in AAV2-R432A eliminated hydrogen bonding interactions, resulting in altered intramolecular and intermolecular interactions propagated from under the 3-fold axis toward the 5-fold channel. Consistent with these observations, differential scanning calorimetry showed an ∼10°C decrease in thermal stability for AAV2-R432A compared to wt-AAV2. In addition, the medium resolution structures revealed differences in the juxtaposition of the less ordered, N-terminal region of their capsid proteins, VP1/2/3. A structural rearrangement in AAV2-R432A repositioned the βA strand region under the icosahedral 2-fold axis rather than antiparallel to the βB strand, eliminating many intramolecular interactions. Thus, a single amino acid substitution can significantly alter the AAV capsid integrity to the extent of reducing its stability and possibly rendering it unable to tolerate the stress of genome packaging. Furthermore, the data show that the 2-, 3-, and 5-fold regions of the capsid contributed to producing the packaging defect and highlight a tight connection between the entire capsid in maintaining packaging efficiency. The mechanism of AAV genome packaging is still poorly understood, particularly with respect to the capsid determinants of the required capsid

A transcriptome survey spanning life stages and sexes of the Harlequin bug, Murgantia histrionica

Science.gov (United States)

The harlequin bug, Murgantia histrionica (Hahn), is an agricultural pest in the continental United States, particularly in southern states. Reliable gene sequence data are especially useful to the development of species-specific, environmentally friendly molecular biopesticides and effective biolure...

No hospital is an island when it comes to Y2K bug

Science.gov (United States)

Silversides, A

1999-01-01

If a recent meeting on the millennium bug is any indication, Canada's health care facilities have plenty to worry about as the year 2,000 approaches. Topics discussed included the state of readiness within the health care sector and potential legal issues that may arise. PMID:9951449

The Impact of Test Case Summaries on Bug Fixing Performance : An Empirical Investigation

NARCIS (Netherlands)

Panichella, S.; Panichella, A.; Beller, M.; Zaidman, A.E.; Gall, H.

2015-01-01

Automated test generation tools have been widely investigated with the goal of reducing the cost of testing activities. However, generated tests have been shown not to help developers in detecting and finding more bugs even though they reach higher structural coverage compared to manual testing. The

The Impact of Test Case Summaries on Bug Fixing Performance : An Empirical Investigation

NARCIS (Netherlands)

Panichella, Sebastiano; Panichella, A.; Beller, M.M.; Zaidman, A.E.; Gall, Harald C.

2016-01-01

Automated test generation tools have been widely investigated with the goal of reducing the cost of testing activities. However, generated tests have been shown not to help developers in detecting and finding more bugs even though they reach higher structural coverage compared to manual testing.

Expression of Aleutian mink disease parvovirus capsid proteins in defined segments: localization of immunoreactive sites and neutralizing epitopes to specific regions.

Science.gov (United States)

Bloom, M E; Martin, D A; Oie, K L; Huhtanen, M E; Costello, F; Wolfinbarger, J B; Hayes, S F; Agbandje-McKenna, M

1997-01-01

The capsid proteins of the ADV-G isolate of Aleutian mink disease parvovirus (ADV) were expressed in 10 nonoverlapping segments as fusions with maltose-binding protein in pMAL-C2 (pVP1, pVP2a through pVP2i). The constructs were designed to capture the VP1 unique sequence and the portions analogous to the four variable surface loops of canine parvovirus (CPV) in individual fragments (pVP2b, pVP2d, pVP2e, and pVP2g, respectively). The panel of fusion proteins was immunoblotted with sera from mink infected with ADV. Seropositive mink infected with either ADV-TR, ADV-Utah, or ADV-Pullman reacted preferentially against certain segments, regardless of mink genotype or virus inoculum. The most consistently immunoreactive regions were pVP2g, pVP2e, and pVP2f, the segments that encompassed the analogs of CPV surface loops 3 and 4. The VP1 unique region was also consistently immunoreactive. These findings indicated that infected mink recognize linear epitopes that localized to certain regions of the capsid protein sequence. The segment containing the hypervariable region (pVP2d), corresponding to CPV loop 2, was also expressed from ADV-Utah. An anti-ADV-G monoclonal antibody and a rabbit anti-ADV-G capsid antibody reacted exclusively with the ADV-G pVP2d segment but not with the corresponding segment from ADV-Utah. Mink infected with ADV-TR or ADV-Utah also preferentially reacted with the pVP2d sequence characteristic of that virus. These results suggested that the loop 2 region may contain a type-specific linear epitope and that the epitope may also be specifically recognized by infected mink. Heterologous antisera were prepared against the VP1 unique region and the four segments capturing the variable surface loops of CPV. The antisera against the proteins containing loop 3 or loop 4, as well as the anticapsid antibody, neutralized ADV-G infectivity in vitro and bound to capsids in immune electron microscopy. These results suggested that regions of the ADV capsid proteins

A new series of HAPs as anti-HBV agents targeting at capsid assembly.

Science.gov (United States)

Yang, Xiu-yan; Xu, Xiao-qian; Guan, Hua; Wang, Li-li; Wu, Qin; Zhao, Guo-ming; Li, Song

2014-09-01

A series of novel Heteroaryldihydropyrimidines (HAPs) derivatives were designed and synthesized as potent inhibitors of HBV capsid assembly. These compounds were prepared from efforts to optimize an earlier series of HAPs, and compounds Mo1, Mo7, Mo8, Mo10, Mo12, and Mo13 demonstrated potent inhibition of HBV DNA replication at submicromolar range. Copyright © 2014. Published by Elsevier Ltd.

Cladistics and biogeography of the assassin bug genus Rasahus Amyot & Serville (Heteroptera: Reduviidae: Peiratinae)

NARCIS (Netherlands)

Morrone, J.J.; Coscarón, M. del C.

1998-01-01

The assassin bug genus Rasahus Amyot & Serville (Heteroptera: Reduviidae: Peiratinae) comprises 26 Neotropical species. A cladistic analysis of the genus was carried out using 63 characters from external morphology, body vestiture, and male and female genitalia, with the species considered as

Side effects of kaolin particle films on apple orchard bug, beetle and spider communities

NARCIS (Netherlands)

Marko, V.; Bogya, S.; Kondorosy, E.; Blommers, L.H.M.

2010-01-01

The effects of multiple applications of hydrophobic kaolin particle film on apple orchard bug (Heteroptera), beetle (Coleoptera) and spider (Araneae) assemblages were studied in the Netherlands. Insecticide-free orchard plots served as a control. The kaolin applications significantly reduced the

First record of Sesbania punicea (Fabales: Fabaceae) as a host plant for Chinavia hilaris (Hemiptera: Pentatomidae)

Science.gov (United States)

The green stink bug, Chinavia hilaris (Say) (Hemiptera: Pentatomidae), is an economic pest of cotton, Gossypium hirsutum L. Numerous known non-crop hosts of C. hilaris that exist in field edges bordering cotton are sources of this stink bug in this crop. Sesbania punicea plants in a field border su...

Studies of gamma emitters in the alluvial deposit and in water of the Bug River

International Nuclear Information System (INIS)

Chibowski, S.; Pawlik, M.

1995-01-01

Studies of the contents of gamma emitters in samples of water and deposit taken from the Bug River (in Poland only) are presented. The total γ-activity of deposits ranges from 120-650 Bq/kg and the average value is 350 Bq/kg. The elevated activity is chiefly due to the presence of natural radionuclides. The main artificial radionuclides are Cs-137 and Cs-134, whose contribution to the total activity ranges from 0.2 to 6%. The activity of waters samples is low, namely, 1.7-3.5 Bq/kg and is due to the presence of natural radionuclides. The heterogeneity of the distribution of natural and artificial radionuclides in the studies samples reveals the effect of economical and industrial activities on contamination of the Bug. (author). 7 refs, 1 fig., 4 tabs

Requirements for capsid-binding and an effector function in TRIMCyp-mediated restriction of HIV-1

International Nuclear Information System (INIS)

Diaz-Griffero, Felipe; Vandegraaff, Nick; Li Yuan; McGee-Estrada, Kathleen; Stremlau, Matthew; Welikala, Sohanya; Si Zhihai; Engelman, Alan; Sodroski, Joseph

2006-01-01

In owl monkeys, a retrotransposition event replaced the gene encoding the retroviral restriction factor TRIM5α with one encoding TRIMCyp, a fusion between the RING, B-box 2 and coiled-coil domains of TRIM5 and cyclophilin A. TRIMCyp restricts human immunodeficiency virus (HIV-1) infection by a mechanism dependent on the interaction of the cyclophilin A moiety and the HIV-1 capsid protein. Here, we show that infection by retroviruses other than HIV-1 can be restricted by TRIMCyp, providing an explanation for the evolutionary retention of the TRIMCyp gene in owl monkey lineages. The TRIMCyp-mediated block to HIV-1 infection occurs before the earliest step of reverse transcription. TRIMCyp-mediated restriction involves at least two functions: (1) capsid binding, which occurs most efficiently for trimeric TRIMCyp proteins that retain the coiled-coil and cyclophilin A domains, and (2) an effector function that depends upon the B-box 2 domain

Gaussian fluctuation of the diffusion exponent of virus capsid in a living cell nucleus

Science.gov (United States)

Itto, Yuichi

2018-05-01

In their work [4], Bosse et al. experimentally showed that virus capsid exhibits not only normal diffusion but also anomalous diffusion in nucleus of a living cell. There, it was found that the distribution of fluctuations of the diffusion exponent characterizing them takes the Gaussian form, which is, quite remarkably, the same form for two different types of the virus. This suggests high robustness of such fluctuations. Here, the statistical property of local fluctuations of the diffusion exponent of the virus capsid in the nucleus is studied. A maximum-entropy-principle approach (originally proposed for a different virus in a different cell) is applied for obtaining the fluctuation distribution of the exponent. Largeness of the number of blocks identified with local areas of interchromatin corrals is also examined based on the experimental data. It is shown that the Gaussian distribution of the local fluctuations can be derived, in accordance with the above form. In addition, it is quantified how the fluctuation distribution on a long time scale is different from the Gaussian distribution.

Fooling the harlequin bug (Hemiptera: Pentatomidae) using synthetic volatiles to alter host plant choice

Science.gov (United States)

Harlequin bug, Murgantia histrionica (Hahn) (Hemiptera: Pentatomidae), is a widespread invasive pest that feeds on a variety of brassicaceous crops and other plants. To understand olfactory cues which mediate host-finding, and possible utility in pest management, we deployed aggregation pheromone (m...

Thermodynamic characterization of the peptide assembly inhibitor binding to HIV-1 capsid protein

Czech Academy of Sciences Publication Activity Database

Kožíšek, Milan; Durčák, Jindřich; Konvalinka, Jan

2013-01-01

Roč. 10, Suppl. 1 (2013), S37-S37 ISSN 1742-4690. [Frontiers of Retrovirology: Complex retorviruses, retroelements and their hosts. 16.09.2013-18.09.2013, Cambridge] R&D Projects: GA ČR GA13-19561S Institutional support: RVO:61388963 Keywords : HIV -1 capsid protein * CAI Subject RIV: EE - Microbiology, Virology http://www.retrovirology.com/content/10/S1/P108

Single Amino Acid Modification of Adeno-Associated Virus Capsid Changes Transduction and Humoral Immune Profiles

Science.gov (United States)

Diprimio, Nina; Bowles, Dawn E.; Hirsch, Matthew L.; Monahan, Paul E.; Asokan, Aravind; Rabinowitz, Joseph; Agbandje-McKenna, Mavis

2012-01-01

Adeno-associated virus (AAV) vectors have the potential to promote long-term gene expression. Unfortunately, humoral immunity restricts patient treatment and in addition provides an obstacle to the potential option of vector readministration. In this study, we describe a comprehensive characterization of the neutralizing antibody (NAb) response to AAV type 1 (AAV1) through AAV5 both in vitro and in vivo. These results demonstrated that NAbs generated from one AAV type are unable to neutralize the transduction of other types. We extended this observation by demonstrating that a rationally engineered, muscle-tropic AAV2 mutant containing 5 amino acid substitutions from AAV1 displayed a NAb profile different from those of parental AAV2 and AAV1. Here we found that a single insertion of Thr from AAV1 into AAV2 capsid at residue 265 preserved high muscle transduction, while also changing the immune profile. To better understand the role of Thr insertion at position 265, we replaced all 20 amino acids and evaluated both muscle transduction and the NAb response. Of these variants, 8 mutants induced higher muscle transduction than AAV2. Additionally, three classes of capsid NAb immune profile were defined based on the ability to inhibit transduction from AAV2 or mutants. While no relationship was found between transduction, amino acid properties, and NAb titer or its cross-reactivity, these studies map a critical capsid motif involved in all steps of AAV infectivity. Our results suggest that AAV types can be utilized not only as templates to generate mutants with enhanced transduction efficiency but also as substrates for repeat administration. PMID:22593151

Facilitating the use of alternative capsid control methods towards sustainable production of organic cocoa in Ghana

NARCIS (Netherlands)

Ayenor, G.K.; Huis, van A.; Obeng-Ofori, D.; Padi, B.; Röling, N.G.

2007-01-01

Cocoa (Theobroma cacao L.) is an important foreign exchange earner for Ghana. However, production is constrained by a high incidence of pests and diseases. Based on farmers' needs, this study focused on the control of capsids, mainly Sahlbergella singularis Haglund and Distantiella theobroma

Transient gene expression in serum-free suspension-growing mammalian cells for the production of foot-and-mouth disease virus empty capsids.

Directory of Open Access Journals (Sweden)

Ana Clara Mignaqui

Full Text Available Foot-and-mouth disease (FMD is a highly contagious disease of cloven-hoofed animals. It produces severe economic losses in the livestock industry. Currently available vaccines are based on inactivated FMD virus (FMDV. The use of empty capsids as a subunit vaccine has been reported to be a promising candidate because it avoids the use of virus in the vaccine production and conserves the conformational epitopes of the virus. In this report, we explored transient gene expression (TGE in serum-free suspension-growing mammalian cells for the production of FMDV recombinant empty capsids as a subunit vaccine. The recombinant proteins produced, assembled into empty capsids and induced protective immune response against viral challenge in mice. Furthermore, they were recognized by anti-FMDV bovine sera. By using this technology, we were able to achieve expression levels that are compatible with the development of a vaccine. Thus, TGE of mammalian cells is an easy to perform, scalable and cost-effective technology for the production of a recombinant subunit vaccine against FMDV.

Field Study of the Comparative Efficacy of Three Pyrethroid/Neonicotinoid Mixture Products for the Control of the Common Bed Bug, Cimex lectularius

Directory of Open Access Journals (Sweden)

Changlu Wang

2015-03-01

Full Text Available Three insecticide mixtures that contain two classes of insecticides (pyrethroid and neonicotinoid were recently developed to control bed bugs. We evaluated three integrated bed bug management strategies in apartments, each using the same non-chemical control methods and one of the three insecticide mixture products: Tandem (lambda-cyhalothrin + thiamethoxam, Temprid SC (beta-cyfluthrin + imidacloprid, and Transport Mikron (bifenthrin + acetamiprid. No insecticides were applied in the Control apartments. In all apartments, we installed vinyl mattress encasements (if not already present and applied steam to beds and other infested upholstered furniture. Insecticide sprays were applied in the three treatments. Each treatment and the Control included 8–10 occupied apartments. Re-treatment was conducted during biweekly inspections if necessary. After eight weeks, the mean (± SEM bed bug count reduction in the Tandem, Temprid SC, Transport Mikron, and Control was 89 ± 9, 87 ± 6, 98 ± 1, and 23 ± 54%, respectively. Only Tandem and Transport Mikron treatments resulted in significantly higher population reduction than the Control at eight weeks. There were no significant differences in mean percent reduction among the three treatments (Tandem, Temprid SC, Transport Mikron at eight weeks. Tandem spray caused significantly faster bed bug reduction than Temprid SC spray and Transport Mikron spray.

Evidence of environmental and vertical transmission of Burkholderia symbionts in the oriental chinch bug, Cavelerius saccharivorus (Heteroptera: Blissidae).

Science.gov (United States)

Itoh, Hideomi; Aita, Manabu; Nagayama, Atsushi; Meng, Xian-Ying; Kamagata, Yoichi; Navarro, Ronald; Hori, Tomoyuki; Ohgiya, Satoru; Kikuchi, Yoshitomo

2014-10-01

The vertical transmission of symbiotic microorganisms is omnipresent in insects, while the evolutionary process remains totally unclear. The oriental chinch bug, Cavelerius saccharivorus (Heteroptera: Blissidae), is a serious sugarcane pest, in which symbiotic bacteria densely populate the lumen of the numerous tubule-like midgut crypts that the chinch bug develops. Cloning and sequence analyses of the 16S rRNA genes revealed that the crypts were dominated by a specific group of bacteria belonging to the genus Burkholderia of the Betaproteobacteria. The Burkholderia sequences were distributed into three distinct clades: the Burkholderia cepacia complex (BCC), the plant-associated beneficial and environmental (PBE) group, and the stinkbug-associated beneficial and environmental group (SBE). Diagnostic PCR revealed that only one of the three groups of Burkholderia was present in ∼89% of the chinch bug field populations tested, while infections with multiple Burkholderia groups within one insect were observed in only ∼10%. Deep sequencing of the 16S rRNA gene confirmed that the Burkholderia bacteria specifically colonized the crypts and were dominated by one of three Burkholderia groups. The lack of phylogenetic congruence between the symbiont and the host population strongly suggested host-symbiont promiscuity, which is probably caused by environmental acquisition of the symbionts by some hosts. Meanwhile, inspections of eggs and hatchlings by diagnostic PCR and egg surface sterilization demonstrated that almost 30% of the hatchlings vertically acquire symbiotic Burkholderia via symbiont-contaminated egg surfaces. The mixed strategy of symbiont transmission found in the oriental chinch bug might be an intermediate stage in evolution from environmental acquisition to strict vertical transmission in insects. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

Detection of Reduced Susceptibility to Chlorfenapyr- and Bifenthrin-Containing Products in Field Populations of the Bed Bug (Hemiptera: Cimicidae).

Science.gov (United States)

Ashbrook, Aaron R; Scharf, Michael E; Bennett, Gary W; Gondhalekar, Ameya D

2017-06-01

Insecticide resistance is a major impediment for effective control of Cimex lectularius L. Previous resistance detection studies with bed bugs have focused on certain pyrethroid, neonicotinoid, organochlorine, organophosphate, and carbamate insecticides. Within the pyrethroid class, resistance studies have mostly been limited to deltamethrin, lambda-cyhalothrin, and alpha- and beta-cyfluthrin. The goal of this study was to develop diagnostic concentration bioassays for assessing bed bug susceptibility levels to chlorfenapyr- and bifenthrin-containing products. First, glass vial and filter paper bioassay methods were compared for their utility in susceptibility monitoring. Statistical comparison of toxicity data between bioassays indicated that the vial assay was less confounded by assay susbtrate effects, required less insecticide, and was faster, especially for chlorfenapyr. Next, using vial diagnostic concentrations (LC99) for each insecticide, 10 laboratory-adapted field strains and the Harlan lab-susceptible strain were screened for susceptibility to chlorfenapyr and bifenthrin. The results of this study reveal recent bed bug susceptibility levels to certain chlorfenapyr- and bifenthrin-containing products. Reduced susceptibility was detected in three and five field strains to chlorfenapyr and bifenthrin, respectively. Detection of reduced susceptibility suggests that certain strains may be segregating toward greater chlorfenapyr and bifenthrin resistance. These results merit continuous resistance monitoring efforts to detect chlorfenapyr and bifenthrin susceptibility shifts. Additionally, to reduce insecticide selection pressures and delay resistance development, adoption of integrated bed bug control strategies that combine chemical and nonchemical methods is recommended. © The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Cold acclimation increases cold tolerance independently of diapause programing in the bean bug, Riptortus pedestris.

Science.gov (United States)

Rozsypal, J; Moos, M; Goto, S G

2017-10-17

The bean bug (Riptortus pedestris) is a pest of soybeans and other legumes in Japan and other Asian countries. It enters a facultative adult diapause on exposure to short days. While photoperiodism and diapause are well understood in R. pedestris, knowledge of cold tolerance is very limited, as is information on the effect of diapause on cold tolerance. We examined the effect of photoperiod, cold acclimation, and feeding status on cold tolerance in R. pedestris. We found that cold acclimation significantly increased survival at -10°C in both long- and short-day adult R. pedestris. Since the difference in cold survival between long- and short-day cold-acclimated groups was only marginal, we conclude that entering diapause is not crucial for R. pedestris to successfully pass through cold acclimation and become cold tolerant. We observed similar effects in 5th instar nymphs, with both long- and short-day cold-acclimated groups surviving longer cold exposures compared with non-acclimated groups. Starvation, which was tested only in adult bugs, had only a negligible and negative impact on cold survival. Although cold tolerance significantly increased with cold acclimation in adult bugs, supercooling capacity unexpectedly decreased. Our results suggest that changes in supercooling capacity as well as in water content are unrelated to cold tolerance in R. pedestris. An analysis of metabolites revealed differences between the treatments, and while several metabolites markedly increased with cold acclimation, their concentrations were too low to have a significant effect on cold tolerance.

Formation of RNA Granule-Derived Capsid Assembly Intermediates Appears To Be Conserved between Human Immunodeficiency Virus Type 1 and the Nonprimate Lentivirus Feline Immunodeficiency Virus.

Science.gov (United States)

Reed, Jonathan C; Westergreen, Nick; Barajas, Brook C; Ressler, Dylan T B; Phuong, Daryl J; Swain, John V; Lingappa, Vishwanath R; Lingappa, Jaisri R

2018-05-01

During immature capsid assembly in cells, human immunodeficiency virus type 1 (HIV-1) Gag co-opts a host RNA granule, forming a pathway of intracellular assembly intermediates containing host components, including two cellular facilitators of assembly, ABCE1 and DDX6. A similar assembly pathway has been observed for other primate lentiviruses. Here we asked whether feline immunodeficiency virus (FIV), a nonprimate lentivirus, also forms RNA granule-derived capsid assembly intermediates. First, we showed that the released FIV immature capsid and a large FIV Gag-containing intracellular complex are unstable during analysis, unlike for HIV-1. We identified harvest conditions, including in situ cross-linking, that overcame this problem, revealing a series of FIV Gag-containing complexes corresponding in size to HIV-1 assembly intermediates. Previously, we showed that assembly-defective HIV-1 Gag mutants are arrested at specific assembly intermediates; here we identified four assembly-defective FIV Gag mutants, including three not previously studied, and demonstrated that they appear to be arrested at the same intermediate as the cognate HIV-1 mutants. Further evidence that these FIV Gag-containing complexes correspond to assembly intermediates came from coimmunoprecipitations demonstrating that endogenous ABCE1 and the RNA granule protein DDX6 are associated with FIV Gag, as shown previously for HIV-1 Gag, but are not associated with a ribosomal protein, at steady state. Additionally, we showed that FIV Gag associates with another RNA granule protein, DCP2. Finally, we validated the FIV Gag-ABCE1 and FIV Gag-DCP2 interactions with proximity ligation assays demonstrating colocalization in situ Together, these data support a model in which primate and nonprimate lentiviruses form intracellular capsid assembly intermediates derived from nontranslating host RNA granules. IMPORTANCE Like HIV-1 Gag, FIV Gag assembles into immature capsids; however, it is not known whether

Biochemical characterisation of the tissue degrading enzyme, collagenase, in the spined soldier bug, Podisus maculiventris (Hemiptera: Pentatomidae

Directory of Open Access Journals (Sweden)

Ghamari Mahboob

2014-07-01

Full Text Available Podisus maculiventris (Say is a generalist predator attacking many insect species from different orders. The bug injects saliva into its prey's body. The ingested hemolymph and liquefied internal tissues pass through the bug's alimentary tract. Collagenase working on peptide bonds of collagen and basement membrane proteins, leads to the disintegration of the prey's internal organs. As yet, there is an almost complete lack of knowledge on the collagenase activity in P. maculiventris. The collagenase activity of the salivary glands and midgut was optimum at pH 8.0 which was congruent with the optimal pH of the total proteolytic activity of the salivary glands. More collagenolytic activity was determined in the posterior lobe of the salivary glands and anterior midgut. Significant inhibition of collagenolytic activity by ethylenediaminetetraacetic acid (EDTA revealed the enzyme is a metalloproteinase. The collagenase activity notably decreased when the bug went hungry. The salivary gland collagenase is a vital enzyme in extra-oral digestion and facilitates the action of other digestive enzymes. The midgut collagenase may be involved in the digestion of the ingested muscle fibers. The collagenase probably acts as an intoxicating agent in the saliva (venom of P. maculiventris. Paralysing toxins are present in the salivary gland secretion.

A case of extensive congregation of Man-faced Stink Bug Catacanthus incarnatus (Drury (Hemiptera: Pentatomidae together with new host records from western Maharashtra, India

Directory of Open Access Journals (Sweden)

S.H. Waghmare

2015-06-01

Full Text Available A study was conducted on the congregation of Stink Bug Catacanthus incarnatus. For the first time this bug was reported at high altitude i.e., 792m. The study reports the congregation of C. incarnatus on four new host plant species viz., Ixora brachiata, Memecylon umbellatum, Glochidion ellipticum and Olea dioica. More infestation was observed on I. brachiata. 

RNA packaging of MRFV virus-like particles: The interplay between RNA pools and capsid coat protein

Science.gov (United States)

Virus-like particles (VLPs) can be produced through self-assembly of capsid protein (CP) into particles with discrete shapes and sizes and containing different types of RNA molecules. The general principle that governs particle assembly and RNA packaging is determined by unique interactions between ...

Use of Cre/loxP recombination to swap cell binding motifs on the adenoviral capsid protein IX

International Nuclear Information System (INIS)

Poulin, Kathy L.; Tong, Grace; Vorobyova, Olga; Pool, Madeline; Kothary, Rashmi; Parks, Robin J.

2011-01-01

We used Cre/loxP recombination to swap targeting ligands present on the adenoviral capsid protein IX (pIX). A loxP-flanked sequence encoding poly-lysine (pK-binds heparan sulfate proteoglycans) was engineered onto the 3'-terminus of pIX, and the resulting fusion protein allowed for routine virus propagation. Growth of this virus on Cre-expressing cells removed the pK coding sequence, generating virus that could only infect through alternative ligands, such as a tyrosine kinase receptor A (TrkA)-binding motif engineered into the capsid fibre protein for enhanced infection of neuronal cells. We used a similar approach to swap the pK motif on pIX for a sequence encoding a single-domain antibody directed towards CD66c for targeted infection of cancer cells; Cre-mediated removal of the pK-coding sequence simultaneously placed the single-domain antibody coding sequence in frame with pIX. Thus, we have developed a simple method to propagate virus lacking native viral tropism but containing cell-specific binding ligands. - Highlights: → We describe a method to grow virus lacking native tropism but containing novel cell-binding ligands. → Cre/loxP recombination was used to modify the adenovirus genome. → A targeting ligand present on capsid protein IX was removed or replaced using recombination. → Cre-loxP was also used to 'swap' the identity of the targeting ligand present on pIX.

The nuclear dimension of the millennium bug: The Syrian experience

International Nuclear Information System (INIS)

Khudeir, I.

1999-01-01

Two nuclear facilities operated by the AECS have been investigated for the Y2K compliance. Computers and associated electronic systems in the Miniature Neutron Source Reactor and the Co-60 irradiator facility were examined. Real time clocks (BIOS) of the personal computers failed Y2K bug and needed to be manually rolled over January 1st 2000. Leap year checks, however were successful. Application software used by the operators for controlling facilities and performing measurements were able to function normally

A Prime-Boost Vaccination Strategy in Cattle to Prevent Foot-and-Mouth Disease Using a "Single-Cycle" Alphavirus Vector and Empty Capsid Particles

DEFF Research Database (Denmark)

Gullberg, Maria; Lohse, Louise; Bøtner, Anette

2016-01-01

Foot-and-mouth disease (FMD) remains one of the most economically important infectious diseases of production animals globally. Vaccination can successfully control this disease, however, current vaccines are imperfect. They are made using chemically inactivated FMD virus (FMDV) that is produced...... in large-scale mammalian cell culture under high containment conditions. Here, we have expressed the FMDV capsid protein precursor (P1-2A) of strain O1 Manisa alone or with the FMDV 3C protease (3Cpro) using a "single cycle" packaged alphavirus self-replicating RNA based on Semliki Forest virus (SFV). When...... the FMDV P1-2A was expressed with 3Cpro then processing of the FMDV capsid precursor protein is observed within cells and the proteins assemble into empty capsid particles. The products interact with anti-FMDV antibodies in an ELISA and bind to the integrin αvβ6 (a cellular receptor for FMDV). In cattle...

Highly specific salt bridges govern bacteriophage P22 icosahedral capsid assembly: identification of the site in coat protein responsible for interaction with scaffolding protein.

Science.gov (United States)

Cortines, Juliana R; Motwani, Tina; Vyas, Aashay A; Teschke, Carolyn M

2014-05-01

Icosahedral virus assembly requires a series of concerted and highly specific protein-protein interactions to produce a proper capsid. In bacteriophage P22, only coat protein (gp5) and scaffolding protein (gp8) are needed to assemble a procapsid-like particle, both in vivo and in vitro. In scaffolding protein's coat binding domain, residue R293 is required for procapsid assembly, while residue K296 is important but not essential. Here, we investigate the interaction of scaffolding protein with acidic residues in the N-arm of coat protein, since this interaction has been shown to be electrostatic. Through site-directed mutagenesis of genes 5 and 8, we show that changing coat protein N-arm residue 14 from aspartic acid to alanine causes a lethal phenotype. Coat protein residue D14 is shown by cross-linking to interact with scaffolding protein residue R293 and, thus, is intimately involved in proper procapsid assembly. To a lesser extent, coat protein N-arm residue E18 is also implicated in the interaction with scaffolding protein and is involved in capsid size determination, since a cysteine mutation at this site generated petite capsids. The final acidic residue in the N-arm that was tested, E15, is shown to only weakly interact with scaffolding protein's coat binding domain. This work supports growing evidence that surface charge density may be the driving force of virus capsid protein interactions. Bacteriophage P22 infects Salmonella enterica serovar Typhimurium and is a model for icosahedral viral capsid assembly. In this system, coat protein interacts with an internal scaffolding protein, triggering the assembly of an intermediate called a procapsid. Previously, we determined that there is a single amino acid in scaffolding protein required for P22 procapsid assembly, although others modulate affinity. Here, we identify partners in coat protein. We show experimentally that relatively weak interactions between coat and scaffolding proteins are capable of driving

Rubella virus capsid protein modulation of viral genomic and subgenomic RNA synthesis

International Nuclear Information System (INIS)

Tzeng, W.-P.; Frey, Teryl K.

2005-01-01

The ratio of the subgenomic (SG) to genome RNA synthesized by rubella virus (RUB) replicons expressing the green fluorescent protein reporter gene (RUBrep/GFP) is substantially higher than the ratio of these species synthesized by RUB (4.3 for RUBrep/GFP vs. 1.3-1.4 for RUB). It was hypothesized that this modulation of the viral RNA synthesis was by one of the virus structural protein genes and it was found that introduction of the capsid (C) protein gene into the replicons as an in-frame fusion with GFP resulted in an increase of genomic RNA production (reducing the SG/genome RNA ratio), confirming the hypothesis and showing that the C gene was the moiety responsible for the modulation effect. The N-terminal one-third of the C gene was required for the effect of be exhibited. A similar phenomenon was not observed with the replicons of Sindbis virus, a related Alphavirus. Interestingly, modulation was not observed when RUBrep/GFP was co-transfected with either other RUBrep or plasmid constructs expressing the C gene, demonstrating that modulation could occur only when the C gene was provided in cis. Mutations that prevented translation of the C protein failed to modulate RNA synthesis, indicating that the C protein was the moiety responsible for modulation; consistent with this conclusion, modulation of RNA synthesis was maintained when synonymous codon mutations were introduced at the 5' end of the C gene that changed the C gene sequence without altering the amino acid sequence of the C protein. These results indicate that C protein translated in proximity of viral replication complexes, possibly from newly synthesized SG RNA, participate in regulating the replication of viral RNA

Green Transformational Leadership and Green Performance: The Mediation Effects of Green Mindfulness and Green Self-Efficacy

Directory of Open Access Journals (Sweden)

Yu-Shan Chen

2014-09-01

Full Text Available No prior literature explores the influence of green transformational leadership on green performance, thus, this study develops a novel research framework to fill the research gap. This study investigates the influence of green transformational leadership on green performance and discusses the mediation effects of green mindfulness and green self-efficacy by means of structural equation modeling (SEM. The results indicate that green transformational leadership positively influences green mindfulness, green self-efficacy, and green performance. Moreover, this study demonstrates that the positive relationship between green transformational leadership and green performance is partially mediated by the two mediators: green mindfulness and green self-efficacy. It means that green transformational leadership can not only directly affect green performance positively but also indirectly affect it positively through green mindfulness and green self-efficacy. Therefore, firms need to raise their green transformational leadership, green mindfulness, and green self-efficacy to increase their green performance.

a study on light trap catches of some rice pests in relation

African Journals Online (AJOL)

Preferred Customer

2 Crop Research Station, Masodha, N.D.U.A. and Technology, Faizabad 224133, India. ABSTRACT: The effect of meteorological factors on the population build up of green leafhopper ... green leafhopper and rice gundhi bug, besides evening ...

TensorCalculator: exploring the evolution of mechanical stress in the CCMV capsid

Science.gov (United States)

Kononova, Olga; Maksudov, Farkhad; Marx, Kenneth A.; Barsegov, Valeri

2018-01-01

A new computational methodology for the accurate numerical calculation of the Cauchy stress tensor, stress invariants, principal stress components, von Mises and Tresca tensors is developed. The methodology is based on the atomic stress approach which permits the calculation of stress tensors, widely used in continuum mechanics modeling of materials properties, using the output from the MD simulations of discrete atomic and C_α -based coarse-grained structural models of biological particles. The methodology mapped into the software package TensorCalculator was successfully applied to the empty cowpea chlorotic mottle virus (CCMV) shell to explore the evolution of mechanical stress in this mechanically-tested specific example of a soft virus capsid. We found an inhomogeneous stress distribution in various portions of the CCMV structure and stress transfer from one portion of the virus structure to another, which also points to the importance of entropic effects, often ignored in finite element analysis and elastic network modeling. We formulate a criterion for elastic deformation using the first principal stress components. Furthermore, we show that von Mises and Tresca stress tensors can be used to predict the onset of a viral capsid’s mechanical failure, which leads to total structural collapse. TensorCalculator can be used to study stress evolution and dynamics of defects in viral capsids and other large-size protein assemblies.

Uncovering Bugs in Distributed Storage Systems during Testing (not in Production!)

OpenAIRE

Deligiannis, P; McCutchen, M; Thomson, P; Chen, S; Donaldson, AF; Erickson, J; Huang, C; Lal, A; Mudduluru, R; Qadeer, S; Schulte, W

2016-01-01

Testing distributed systems is challenging due to multiple sources of nondeterminism. Conventional testing techniques, such as unit, integration and stress testing, are ineffective in preventing serious but subtle bugs from reaching production. Formal techniques, such as TLA+, can only verify high-level specifications of systems at the level of logic-based models, and fall short of checking the actual executable code. In this paper, we present a new methodology for testing distributed systems...

Capsid coding sequences of foot-and-mouth disease viruses are determinants of pathogenicity in pigs.

Science.gov (United States)

Lohse, Louise; Jackson, Terry; Bøtner, Anette; Belsham, Graham J

2012-05-24

The surface exposed capsid proteins, VP1, VP2 and VP3, of foot-and-mouth disease virus (FMDV) determine its antigenicity and the ability of the virus to interact with host-cell receptors. Hence, modification of these structural proteins may alter the properties of the virus.In the present study we compared the pathogenicity of different FMDVs in young pigs. In total 32 pigs, 7-weeks-old, were exposed to virus, either by direct inoculation or through contact with inoculated pigs, using cell culture adapted (O1K B64), chimeric (O1K/A-TUR and O1K/O-UKG) or field strain (O-UKG/34/2001) viruses. The O1K B64 virus and the two chimeric viruses are identical to each other except for the capsid coding region.Animals exposed to O1K B64 did not exhibit signs of disease, while pigs exposed to each of the other viruses showed typical clinical signs of foot-and-mouth disease (FMD). All pigs infected with the O1K/O-UKG chimera or the field strain (O-UKG/34/2001) developed fulminant disease. Furthermore, 3 of 4 in-contact pigs exposed to the O1K/O-UKG virus died in the acute phase of infection, likely from myocardial infection. However, in the group exposed to the O1K/A-TUR chimeric virus, only 1 pig showed symptoms of disease within the time frame of the experiment (10 days). All pigs that developed clinical disease showed a high level of viral RNA in serum and infected pigs that survived the acute phase of infection developed a serotype specific antibody response. It is concluded that the capsid coding sequences are determinants of FMDV pathogenicity in pigs.

Generation of neutralizing monoclonal antibodies against a conformational epitope of human adenovirus type 7 (HAdv-7 incorporated in capsid encoded in a HAdv-3-based vector.

Directory of Open Access Journals (Sweden)

Minglong Liu

Full Text Available The generation of monoclonal antibodies (MAbs by epitope-based immunization is difficult because the immunogenicity of simple peptides is poor and T cells must be potently stimulated and immunological memory elicited. A strategy in which antigen is incorporated into the adenoviral capsid protein has been used previously to develop antibody responses against several vaccine targets and may offer a solution to this problem. In this study, we used a similar strategy to develop HAdv-7-neutralizing MAbs using rAdMHE3 virions into which hexon hypervariable region 5 (HVR5 of adenovirus type 7 (HAdv-7 was incorporated. The epitope mutant rAdMHE3 was generated by replacing HVR5 of Ad3EGFP, a recombinant HAdv-3-based vector expressing enhanced green fluorescence protein, with HVR5 of HAdv-7. We immunized BALB/c mice with rAdMHE3 virions and produced 22 different MAbs against them, four of which showed neutralizing activity against HAdv-7 in vitro. Using an indirect enzyme-linked immunosorbent assay (ELISA analysis and an antibody-binding-competition ELISA with Ad3EGFP, HAdv-7, and a series of chimeric adenoviral particles containing epitope mutants, we demonstrated that the four MAbs recognize the neutralization site within HVR5 of the HAdv-7 virion. Using an immunoblotting analysis and ELISA with HAdv-7, recombinant peptides, and a synthetic peptide, we also showed that the neutralizing epitope within HVR5 of the HAdv-7 virion is a conformational epitope. These findings suggest that it is feasible to use a strategy in which antigen is incorporated into the adenoviral capsid protein to generate neutralizing MAbs. This strategy may also be useful for developing therapeutic neutralizing MAbs and designing recombinant vector vaccines against HAdv-7, and in structural analysis of adenoviruses.

BI-2 destabilizes HIV-1 cores during infection and Prevents Binding of CPSF6 to the HIV-1 Capsid.

Science.gov (United States)

Fricke, Thomas; Buffone, Cindy; Opp, Silvana; Valle-Casuso, Jose; Diaz-Griffero, Felipe

2014-12-11

The recently discovered small-molecule BI-2 potently blocks HIV-1 infection. BI-2 binds to the N-terminal domain of HIV-1 capsid. BI-2 utilizes the same capsid pocket used by the small molecule PF74. Although both drugs bind to the same pocket, it has been proposed that BI-2 uses a different mechanism to block HIV-1 infection when compared to PF74. This work demonstrates that BI-2 destabilizes the HIV-1 core during infection, and prevents the binding of the cellular factor CPSF6 to the HIV-1 core. Overall this short-form paper suggests that BI-2 is using a similar mechanism to the one used by PF74 to block HIV-1 infection.

Apocalypse deterred : oil and gas community pulls out ahead in vaccinating itself against Y2K, the millennium bug

International Nuclear Information System (INIS)

Gordon, M.

1999-01-01

Results of a survey of the membership of the Canadian Association of Petroleum Producers (CAPP) regarding efforts to deal with the threat posed by the Y2K millennium bug are reviewed. Survey results reveal that CAPP's 170 member companies take the year 2000 date rollover very seriously, and the vast majority (99 per cent) have formal action plans to deal with the threat. The vast majority of plans have components that take into account the effects of the computer glitch on inventories, compliance and risk assessment matters, correction and replacement. All are doing testing and assessments involving business partners, and are developing contingency plans to keep operating if any problems develop despite all the efforts currently underway. Test results to date show low failure rates in field production control equipment. The current focus is on contingency plans for higher risk items. The downstream sector of the industry, involving transportation, distribution and retail marketing, is making similar efforts to minimize the threat of the Y2K bug, sometimes to the point of switching to new and improved equipment to vaccinate all its major operations against the expected computer failure. The cost of coping with the millennium bug is colossal. Canadian governments expect the cost to reach three billion dollars to purge the bug from government operations alone. Estimates of world-wide cost run to 530 billion dollars, plus another trillion dollars to clean up the aftermath of infections. Nevertheless, the current assessment is that the back of the problem has been broken at least in Canada, the United States and Western Europe. Nevertheless, there is no justification for complacency just yet

Structure of the immature HIV-1 capsid in intact virus particles at 8.8 angstrom resolution

Czech Academy of Sciences Publication Activity Database

Schur, F. K. M.; Hagen, W. J. H.; Rumlová, Michaela; Ruml, T.; Müller, B.; Kräusslich, H. G.; Briggs, J. A. G.

2015-01-01

Roč. 517, č. 7535 (2015), s. 505-508 ISSN 0028-0836 R&D Projects: GA ČR(CZ) GA14-15326S Institutional support: RVO:61388963 Keywords : retrovirus * HIV * M-PMV * capsid protein * CA * assembly * immature particles Subject RIV: CE - Biochemistry Impact factor: 38.138, year: 2015

Characterization of Halyomorpha halys (brown marmorated stink bug) biogenic volatile organic compound emissions and their role in secondary organic aerosol formation.

Science.gov (United States)

Solomon, Danielle; Dutcher, Dabrina; Raymond, Timothy

2013-11-01

The formation of aerosols is a key component in understanding cloud formation in the context of radiative forcings and global climate modeling. Biogenic volatile organic compounds (BVOCs) are a significant source of aerosols, yet there is still much to be learned about their structures, sources, and interactions. The aims of this project were to identify the BVOCs found in the defense chemicals of the brown marmorated stink bug Halymorpha halys and quantify them using gas chromatography-mass spectrometry (GC/MS) and test whether oxidation of these compounds by ozone-promoted aerosol and cloud seed formation. The bugs were tested under two conditions: agitation by asphyxiation and direct glandular exposure. Tridecane, 2(5H)-furanone 5-ethyl, and (E)-2-decenal were identified as the three most abundant compounds. H. halys were also tested in the agitated condition in a smog chamber. It was found that in the presence of 100-180 ppm ozone, secondary aerosols do form. A scanning mobility particle sizer (SMPS) and a cloud condensation nuclei counter (CCNC) were used to characterize the secondary aerosols that formed. This reaction resulted in 0.23 microg/ bug of particulate mass. It was also found that these secondary organic aerosol particles could act as cloud condensation nuclei. At a supersaturation of 1%, we found a kappa value of 0.09. Once regional populations of these stink bugs stablilize and the populations estimates can be made, the additional impacts of their contribution to regional air quality can be calculated.

Doing bayesian data analysis a tutorial with R and BUGS

CERN Document Server

Kruschke, John K

2011-01-01

There is an explosion of interest in Bayesian statistics, primarily because recently created computational methods have finally made Bayesian analysis obtainable to a wide audience. Doing Bayesian Data Analysis, A Tutorial Introduction with R and BUGS provides an accessible approach to Bayesian data analysis, as material is explained clearly with concrete examples. The book begins with the basics, including essential concepts of probability and random sampling, and gradually progresses to advanced hierarchical modeling methods for realistic data. The text delivers comprehensive coverage of all

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

Directory of Open Access Journals (Sweden)

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

Enterovirus 71 viral capsid protein linear epitopes: Identification and characterization

Directory of Open Access Journals (Sweden)

Gao Fan

2012-01-01

Full Text Available Abstract Background To characterize the human humoral immune response against enterovirus 71 (EV71 infection and map human epitopes on the viral capsid proteins. Methods A series of 256 peptides spanning the capsid proteins (VP1, VP2, VP3 of BJ08 strain (genomic C4 were synthesized. An indirect enzyme-linked immunosorbent assay (ELISA was carried out to detect anti-EV71 IgM and IgG in sera of infected children in acute or recovery phase. The partially overlapped peptides contained 12 amino acids and were coated in the plate as antigen (0.1 μg/μl. Sera from rabbits immunized with inactivated BJ08 virus were also used to screen the peptide panel. Results A total of 10 human anti-EV71 IgM epitopes (vp1-14 in VP1; vp2-6, 21, 40 and 50 in VP2 and vp3-10, 12, 15, 24 and 75 in VP3 were identified in acute phase sera. In contrast, only one anti-EV71 IgG epitope in VP1 (vp1-15 was identified in sera of recovery stage. Four rabbit anti-EV71 IgG epitopes (vp1-14, 31, 54 and 71 were identified and mapped to VP1. Conclusion These data suggested that human IgM epitopes were mainly mapped to VP2 and VP3 with multi-epitope responses occurred at acute infection, while the only IgG epitope located on protein VP1 was activated in recovery phase sera. The dynamic changes of humoral immune response at different stages of infection may have public health significance in evaluation of EV71 vaccine immunogenicity and the clinical application of diagnostic reagents.

Characterization of a protein kinase activity associated with purified capsids of the granulosis virus infecting Plodia interpunctella.

Science.gov (United States)

Wilson, M E; Consigli, R A

1985-06-01

A cyclic-nucleotide independent protein kinase activity has been demonstrated in highly purified preparations of the granulosis virus infecting the Indian meal moth, Plodia interpunctella. A divalent cation was required for activity. Manganese was the preferred cation and a pH of 8.0 resulted in optimal incorporation of 32P radiolabel into acid-precipitable protein. Although both ATP and GTP could serve as phosphate donors, ATP was utilized more efficiently by the enzyme. The kinase activity was localized to purified capsids; and the basic, internal core protein, VP12, was found to be the predominant viral acceptor. Histones and protamine sulfate could also serve as acceptors for the capsid-associated kinase activity. Using acid hydrolysis and phosphoamino acid analysis of phosphorylated nucleocapsid protein and nuclear magnetic resonance of phosphorylated VP12, it was determined that the enzyme catalyzes the transfer of phosphate to both serine and arginine residues of acceptor proteins. We believe this kinase activity may play a significant role in the viral replication cycle.

Green Transformational Leadership and Green Performance: The Mediation Effects of Green Mindfulness and Green Self-Efficacy

OpenAIRE

Yu-Shan Chen; Ching-Hsun Chang; Yu-Hsien Lin

2014-01-01

No prior literature explores the influence of green transformational leadership on green performance, thus, this study develops a novel research framework to fill the research gap. This study investigates the influence of green transformational leadership on green performance and discusses the mediation effects of green mindfulness and green self-efficacy by means of structural equation modeling (SEM). The results indicate that green transformational leadership positively influences green min...

An insight into the sialotranscriptome of the seed-feeding bug, Oncopeltus fasciatus

OpenAIRE

Francischetti, Ivo M.B.; Lopes, Angela H.; Dias, Felipe A.; Pham, Van M.; Ribeiro, José M.C.

2007-01-01

The salivary transcriptome of the seed-feeding hemipteran, Oncopeltus fasciatus (milkweed bug), is described following assembly of 1,025 ESTs into 305 clusters of related sequences. Inspection of these sequences reveals abundance of low complexity, putative secreted products rich in the amino acids (aa) glycine, serine or threonine, which might function as silk or mucins and assist food canal lubrication and sealing of the feeding site around the mouthparts. Several protease inhibitors were f...

New Introductions, Spread of Existing Matrilines, and High Rates of Pyrethroid Resistance Result in Chronic Infestations of Bed Bugs (Cimex lectularius L. in Lower-Income Housing.

Directory of Open Access Journals (Sweden)

Ronald W Raab

Full Text Available Infestations of the common bed bug (Cimex lectularius L. have increased substantially in the United States in the past 10-15 years. The housing authority in Harrisonburg, Virginia, conducts heat-treatments after bed bugs are detected in a lower-income housing complex, by treating each infested unit at 60°C for 4-6 hours. However, a high frequency of recurrent infestations called into question the efficacy of this strategy. Genetic analysis using Bayesian clustering of polymorphic microsatellite loci from 123 bed bugs collected from 23 units from May 2012 to April 2013 in one building indicated that (a 16/21 (73% infestations were genetically similar, suggesting ineffective heat-treatments or reintroductions from within the building or from a common external source, followed by local spread of existing populations; and (b up to 5 of the infestations represented new genotypes, indicating that 5 new populations were introduced into this building in one year, assuming they were not missed in earlier screens. There was little to no gene flow among the 8 genetic clusters identified in the building. Bed bugs in the U.S. often possess one or both point mutations in the voltage-gated sodium channel, termed knockdown resistance (kdr, from valine to leucine (V419L and leucine to isoleucine (L925I that confer target-site resistance against pyrethroid insecticides. We found that 48/121 (40% bed bugs were homozygous for both kdr mutations (L419/I925, and a further 59% possessed at least one of the kdr mutations. We conclude that ineffective heat treatments, new introductions, reintroductions and local spread, and an exceptionally high frequency of pyrethroid resistance are responsible for chronic infestations in lower-income housing. Because heat treatments fail to protect from reintroductions, and pesticide use has not decreased the frequency of infestations, preventing new introductions and early detection are the most effective strategies to avoid bed bug

GREEN PACKAGING, GREEN PRODUCT, GREEN ADVERTISING, PERSEPSI, DAN MINAT BELI KONSUMEN

Directory of Open Access Journals (Sweden)

Imam Santoso

2016-12-01

Full Text Available Environmental problems become one of the strategic issues in achieving global competitiveness. One of the issues is products that are made from environmental friendly materials or known as green product. Furthermore, in green products marketing, the company also uses green packaging and green advertising concept. This study aimed to analyze the effect of green packaging, green products, and green advertising on consumer perception and purchasing intention. The study was conducted in Ketawanggede Village, Lowokwaru Sub-district, Malang City. The sampling method used nonprobability accidential sampling techniques. The numbers of respondents were 113 consumers in study site. Data were collected by interview using questionnaires. The method of analysis used Generalized Structured Component Analysis (GSCA. The analysis showed that the green packaging, green products, and green advertising had positive significant influence on consumer perceptions. Meanwhile, green product and consumer perception had positive significant influence on purchasing interest, but the green packaging and green advertising has not found sufficient evidence in influencing purchasing intention.

RNAi analysis of Deformed, proboscipedia and Sex combs reduced in the milkweed bug Oncopeltus fasciatus: novel roles for Hox genes in the hemipteran head.

Science.gov (United States)

Hughes, C L; Kaufman, T C

2000-09-01

Insects have evolved a large variety of specialized feeding strategies, with a corresponding variability in mouthpart morphology. We have, however, little understanding of the developmental mechanisms that underlie this diversity. Until recently it was difficult to perform any analysis of gene function outside of the genetic model insects Drosophila melanogaster and Tribolium castaneum. In this paper, we report the use of dsRNA-mediated interference (RNAi) to dissect gene function in the development of the milkweed bug Oncopeltus fasciatus, which has specialized suctorial mouthparts. The Hox genes Deformed (Dfd), proboscipedia (pb) and Sex combs reduced (Scr) have previously been shown to be expressed in the gnathal appendages of this species. Strikingly, the milkweed bug was found to have an unusual expression pattern of pb. Here, by analyzing single and combination RNAi depletions, we find that Dfd, pb and Scr are used in the milkweed bug to specify the identity of the mouthparts. The exact roles of the genes, however, are different from what is known in the two genetic model insects. The maxillary appendages in the bug are determined by the activities of the genes Dfd and Scr, rather than Dfd and pb as in the fly and beetle. The mandibular appendages are specified by Dfd, but their unique morphology in Oncopeltus suggests that Dfd's target genes are different. As in flies and beetles, the labium is specified by the combined activities of pb and Scr, but again, the function of pb appears to be different. Additionally, the regulatory control of pb by the other two genes seems to be different in the bug than in either of the other species. These novelties in Hox function, expression pattern and regulatory relationships may have been important for the evolution of the unique Hemipteran head.

Structures of the major capsid proteins of the human Karolinska Institutet and Washington University polyomaviruses.

Science.gov (United States)

Neu, Ursula; Wang, Jianbo; Macejak, Dennis; Garcea, Robert L; Stehle, Thilo

2011-07-01

The Karolinska Institutet and Washington University polyomaviruses (KIPyV and WUPyV, respectively) are recently discovered human viruses that infect the respiratory tract. Although they have not yet been linked to disease, they are prevalent in populations worldwide, with initial infection occurring in early childhood. Polyomavirus capsids consist of 72 pentamers of the major capsid protein viral protein 1 (VP1), which determines antigenicity and receptor specificity. The WUPyV and KIPyV VP1 proteins are distant in evolution from VP1 proteins of known structure such as simian virus 40 or murine polyomavirus. We present here the crystal structures of unassembled recombinant WUPyV and KIPyV VP1 pentamers at resolutions of 2.9 and 2.55 Å, respectively. The WUPyV and KIPyV VP1 core structures fold into the same β-sandwich that is a hallmark of all polyomavirus VP1 proteins crystallized to date. However, differences in sequence translate into profoundly different surface loop structures in KIPyV and WUPyV VP1 proteins. Such loop structures have not been observed for other polyomaviruses, and they provide initial clues about the possible interactions of these viruses with cell surface receptors.

Comparative Mitogenomics of Plant Bugs (Hemiptera: Miridae): Identifying the AGG Codon Reassignments between Serine and Lysine

Science.gov (United States)

Wang, Pei; Song, Fan; Cai, Wanzhi

2014-01-01

Insect mitochondrial genomes are very important to understand the molecular evolution as well as for phylogenetic and phylogeographic studies of the insects. The Miridae are the largest family of Heteroptera encompassing more than 11,000 described species and of great economic importance. For better understanding the diversity and the evolution of plant bugs, we sequence five new mitochondrial genomes and present the first comparative analysis of nine mitochondrial genomes of mirids available to date. Our result showed that gene content, gene arrangement, base composition and sequences of mitochondrial transcription termination factor were conserved in plant bugs. Intra-genus species shared more conserved genomic characteristics, such as nucleotide and amino acid composition of protein-coding genes, secondary structure and anticodon mutations of tRNAs, and non-coding sequences. Control region possessed several distinct characteristics, including: variable size, abundant tandem repetitions, and intra-genus conservation; and was useful in evolutionary and population genetic studies. The AGG codon reassignments were investigated between serine and lysine in the genera Adelphocoris and other cimicomorphans. Our analysis revealed correlated evolution between reassignments of the AGG codon and specific point mutations at the antidocons of tRNALys and tRNASer(AGN). Phylogenetic analysis indicated that mitochondrial genome sequences were useful in resolving family level relationship of Cimicomorpha. Comparative evolutionary analysis of plant bug mitochondrial genomes allowed the identification of previously neglected coding genes or non-coding regions as potential molecular markers. The finding of the AGG codon reassignments between serine and lysine indicated the parallel evolution of the genetic code in Hemiptera mitochondrial genomes. PMID:24988409

Computer games to teach hygiene: an evaluation of the e-Bug junior game

OpenAIRE

Farrell, D.; Kostkova, P.; Weinberg, J.; Lazareck, L.; Weerasinghe, D.; Lecky, D. M.; McNulty, C. A.

2011-01-01

Handwashing, respiratory hygiene and antibiotic resistance remain major public health concerns. In order to facilitate an effective outcome when teaching the basic principles of hand and respiratory hygiene, educational interventions should first target school children. As computer games are ubiquitous in most children's lives, e-Bug developed computer games targeted at teaching children handwashing, respiratory hygiene and antibiotic resistance. The games were designed for two target audienc...

Simulating the 2012 High Plains drought using three single column versions (SCM) of BUGS5

Science.gov (United States)

Medina, I. D.; Denning, S.

2013-12-01

The impact of changes in the frequency and severity of drought on fresh water sustainability is a great concern for many regions of the world. One such location is the High Plains, where the local economy is primarily driven by fresh water withdrawals from the Ogallala Aquifer, which accounts for approximately 30% of total irrigation withdrawals from all U.S. aquifers combined. Modeling studies that focus on the feedback mechanisms that control the climate and eco-hydrology during times of drought are limited, and have used conventional General Circulation Models (GCMs) with grid length scales ranging from one hundred to several hundred kilometers. Additionally, these models utilize crude statistical parameterizations of cloud processes for estimating sub-grid fluxes of heat and moisture and have a poor representation of land surface heterogeneity. For this research, we will focus on the 2012 High Plains drought and will perform numerical simulations using three single column versions (SCM) of BUGS5 (Colorado State University (CSU) GCM coupled to the Simple Biosphere Model (SiB3)) at multiple sites overlying the Ogallala Aquifer for the 2011-2012 periods. In the first version of BUGS5, the model will be used in its standard bulk setting (single atmospheric column coupled to a single instance of SiB3), secondly, the Super-Parameterized Community Atmospheric Model (SP-CAM), a cloud resolving model (CRM consists of 64 atmospheric columns), will replace the single CSU GCM atmospheric parameterization and will be coupled to a single instance of SiB3, and for the third version of BUGS5, an instance of SiB3 will be coupled to each CRM column of the SP-CAM (64 CRM columns coupled to 64 instances of SiB3). To assess the physical realism of the land-atmosphere feedbacks simulated at each site by all versions of BUGS5, differences in simulated energy and moisture fluxes will be computed between the 2011 and 2012 period and will be compared to differences calculated using

Unveiling Exception Handling Bug Hazards in Android Based on GitHub and Google Code Issues

NARCIS (Netherlands)

Coelho, R.; Almeida, L.; Gousios, G.; Van Deursen, A.

2015-01-01

This paper reports on a study mining the exception stack traces included in 159,048 issues reported on Android projects hosted in GitHub (482 projects) and Google Code (157 projects). The goal of this study is to investigate whether stack trace information can reveal bug hazards related to exception

Expression and purification of capsid proteins of Aichi virus and in vitro reassembly of empty virion

Czech Academy of Sciences Publication Activity Database

Smola, Miroslav; Dubánková, Anna; Šilhán, Jan; Bouřa, Evžen

2017-01-01

Roč. 284, Suppl 1 (2017), s. 107 ISSN 1742-464X. [FEBS Congress /42./ From Molecules to Cells and Back. 10.09.2017-14.09.2017, Jerusalem] R&D Projects: GA ČR GJ15-21030Y; GA MŠk LO1302 Institutional support: RVO:61388963 Keywords : Aichi virus * capsid proteins Subject RIV: CE - Biochemistry

Laboratory efficacy of mycoparasitic fungi (Aspergillus tubingensis and Trichoderma harzianum against tropical bed bugs (Cimex hemipterus (Hemiptera: Cimicidae

Directory of Open Access Journals (Sweden)

Zulaikha Zahran

2017-04-01

Conclusions: Approaches to the bed bugs treatment should be explored in-depth using a natural biological agent like fungus especially A. tubingensis to reduce this pest population, in order to replace chemical methods.

Milkweed (Gentianales: Apocynaceae): a farmscape resource for increasing parasitism of stink bugs (Hemiptera: Pentatomidae) and providing nectar to insect pollinators and monarch butterflies.

Science.gov (United States)

Tillman, P G; Carpenter, J E

2014-04-01

In peanut-cotton farmscapes in Georgia, the stink bugs Nezara viridula (L.) and Chinavia hilaris (Say) (Hemiptera: Pentatomidae) and the leaffooted bug, Leptoglossus phyllopus (L.) (Hemiptera: Coreidae), disperse at crop-to-crop interfaces to feed on bolls in cotton. The main objective of this study was to determine whether insecticide-free tropical milkweed (Asclepias curassavica L.), a nectar-producing plant, can increase parasitism of these bugs by Trichopoda pennipes (F.) (Diptera: Tachinidae) and provide nectar to monarch butterflies and insect pollinators in these farmscapes. Peanut-cotton plots with and without flowering milkweed plants were established in 2009 and 2010. Adult T. pennipes, monarch butterflies, honey bees, and native insect pollinators readily fed on floral nectar of milkweed. Monarch larvae feeding on milkweed vegetation successfully developed into pupae. In 2009, N. viridula was the primary host of T. pennipes in cotton, and parasitism of this pest by the parasitoid was significantly higher in milkweed cotton (61.6%) than in control cotton (13.3%). In 2010, parasitism of N. viridula, C. hilaris, and L. phyllopus by T. pennipes was significantly higher in milkweed cotton (24.0%) than in control cotton (1.1%). For both years of the study, these treatment differences were not owing to a response by the parasitoid to differences in host density, because density of hosts was not significantly different between treatments. In conclusion, incorporation of milkweed in peanut-cotton plots increased stink bug parasitism in cotton and provided nectar to insect pollinators and monarch butterflies.

Preventing Run-Time Bugs at Compile-Time Using Advanced C++

Energy Technology Data Exchange (ETDEWEB)

Neswold, Richard [Fermilab

2018-01-01

When writing software, we develop algorithms that tell the computer what to do at run-time. Our solutions are easier to understand and debug when they are properly modeled using class hierarchies, enumerations, and a well-factored API. Unfortunately, even with these design tools, we end up having to debug our programs at run-time. Worse still, debugging an embedded system changes its dynamics, making it tough to find and fix concurrency issues. This paper describes techniques using C++ to detect run-time bugs *at compile time*. A concurrency library, developed at Fermilab, is used for examples in illustrating these techniques.

GREEN PACKAGING, GREEN PRODUCT, GREEN ADVERTISING, PERSEPSI, DAN MINAT BELI KONSUMEN

OpenAIRE

Imam Santoso; Rengganis Fitriani

2016-01-01

Environmental problems become one of the strategic issues in achieving global competitiveness. One of the issues is products that are made from environmental friendly materials or known as green product. Furthermore, in green products marketing, the company also uses green packaging and green advertising concept. This study aimed to analyze the effect of green packaging, green products, and green advertising on consumer perception and purchasing intention. The study was conducted in Ketawangg...

Recovering stereo vision by squashing virtual bugs in a virtual reality environment

OpenAIRE

Vedamurthy, I; Knill, DC; Huang, SJ; Yung, A; Ding, J; Kwon, OS; Bavelier, D; Levi, DM

2016-01-01

Stereopsis is the rich impression of three-dimensionality, based on binocular disparity—the differences between the two retinal images of the same world. However, a substantial proportion of the population is stereo-deficient, and reliesmostly onmonocular cues to judge the relative depth or distance of objects in the environment. Here we trained adults who were stereo blind or stereodeficient owing to strabismus and/or amblyopia in a natural visuomotor task—a ‘bug squashing’ game—in a virtual...

The lectin from Musa paradisiaca binds with the capsid protein of tobacco mosaic virus and prevents viral infection.

Science.gov (United States)

Liu, Xiao-Yu; Li, Huan; Zhang, Wei

2014-05-04

It has been demonstrated that the lectin from Musa paradisiaca (BanLec-1) could inhibit the cellular entry of human immunodeficiency virus (HIV). In order to evaluate its effects on tobacco mosaic virus (TMV), the banlec-1 gene was cloned and transformed into Escherichia coli and tobacco, respectively. Recombinant BanLec-1 showed metal ions dependence, and higher thermal and pH stability. Overexpression of banlec-1 in tobacco resulted in decreased leaf size, and higher resistance to TMV infection, which includes reduced TMV cellular entry, more stable chlorophyll contents, and enhanced antioxidant enzymes. BanLec-1 was found to bind directly to the TMV capsid protein in vitro , and to inhibit TMV infection in a dose-dependent manner. In contrast to limited prevention in vivo , purified rBanLec-1 exhibited more significant effects on TMV infection in vitro . Taken together, our study indicated that BanLec-1 could prevent TMV infection in tobacco, probably through the interaction between BanLec-1 and TMV capsid protein.

Biological control program is being developed for brown marmorated stink bug

OpenAIRE

Lara, Jesus; Pickett, Charlie; Ingels, Chuck; Haviland, David R; Grafton-Cardwell, Elizabeth E; Doll, David A; Bethke, James A; Faber, Ben; Dara, Surendra K; Hoddle, Mark

2016-01-01

Brown marmorated stink bug (BMSB) is an invasive, polyphagous pest that has been detected in 42 U.S. states. In 2010, it caused millions of dollars in crop damages to apple growers on the East Coast, where it arrived from Asia during the 1990s. In 2002, BMSB was reported in California; since then, it has been detected in 28 counties and is established in at least nine counties. Although this pest has not yet been found on commercial crops in the state, detections of BMSB in commercial orchard...

Millennial angst? Insidious Y2K computer bug could have devastating consequences if not remedied

International Nuclear Information System (INIS)

Ross, E.

1999-01-01

The threat of the Y2K bug facing the petroleum industry was discussed. The bug can potentially affect every aspect of the industry relying on computerized and automated systems and equipment, from production to transportation. The importance for oil companies to establish contingency plans was emphasized. For example, Enbridge Energy Inc. is planning to be Y2K ready on all mission critical systems by June 1999. On the pipeline side, this means anything dealing with physical safety and the operational reliability of the pipeline. Canadian Occidental Petroleum Ltd., has tested everything dealing with environmental or public safety risk and regularly checks on how their suppliers are progressing with Y2K compliance. Companies will also face regulatory requirements, For example, the Alberta Energy and Utilities Board will be checking on 127 companies considered high risk, primarily for safety reasons. This includes operators of sour gas plants or pipelines and gas or electrical utilities. The upstream petroleum industry generally, is reported to be making good progress in addressing the issue

Greens of the European Green Capitals

Science.gov (United States)

Cömertler, Seval

2017-10-01

Well established and maintained green areas have a key role on reaching the high quality of life and sustainability in urban environments. Therefore, green areas must be carefully accounted and evaluated in the urban planning affairs. In this context, the European Green Capitals, which attach a great importance to the green areas, have a great potential to act as a role model for both small and big cities in all around the world. These leading cities (chronologically, Stockholm, Hamburg, Vitoria-Gasteiz, Nantes, Copenhagen, Bristol, Ljubljana, Essen and Nijmegen) are inspiring for the other cities which seek to achieve more sustainable and environmentally friendly places through green areas. From this point of view, the aim of this paper was to investigate the green areas of the European Green Capitals. The paper covered whole European Green Capitals, and the application form of each Green Capital was used as a primary data source. Consequently, the paper put forwarded that the European Green Capitals have considerably large amount and high proportion of green areas. Further, these cities provide an excellent access to the public green areas. As a result of abundant provision and proper distribution, the almost all citizens in most of the Green Capitals live within a distance of 300 meters to a green area. For further researches, the paper suggested that these green capitals should be investigated in terms of their efforts, measures, goals and plans, policies and implications to administer, to protect, to enhance and to expand the green areas.

How Do Visitors Relate to Biodiversity Conservation? An Analysis of London Zoo's "BUGS" Exhibit

Science.gov (United States)

Chalmin-Pui, Lauriane Suyin; Perkins, Richard

2017-01-01

Using a case study of London Zoo's BUGS (Biodiversity Underpinning Global Survival) exhibit, this article assesses the role of experiential learning in raising biodiversity knowledge, concern and potential pro-conservation actions. Using Personal Meaning Mindmapping, a novel method in visitor research, the study examines how adult visitors relate…

Recovering stereo vision by squashing virtual bugs in a virtual reality environment.

Science.gov (United States)

Vedamurthy, Indu; Knill, David C; Huang, Samuel J; Yung, Amanda; Ding, Jian; Kwon, Oh-Sang; Bavelier, Daphne; Levi, Dennis M

2016-06-19

Stereopsis is the rich impression of three-dimensionality, based on binocular disparity-the differences between the two retinal images of the same world. However, a substantial proportion of the population is stereo-deficient, and relies mostly on monocular cues to judge the relative depth or distance of objects in the environment. Here we trained adults who were stereo blind or stereo-deficient owing to strabismus and/or amblyopia in a natural visuomotor task-a 'bug squashing' game-in a virtual reality environment. The subjects' task was to squash a virtual dichoptic bug on a slanted surface, by hitting it with a physical cylinder they held in their hand. The perceived surface slant was determined by monocular texture and stereoscopic cues, with these cues being either consistent or in conflict, allowing us to track the relative weighting of monocular versus stereoscopic cues as training in the task progressed. Following training most participants showed greater reliance on stereoscopic cues, reduced suppression and improved stereoacuity. Importantly, the training-induced changes in relative stereo weights were significant predictors of the improvements in stereoacuity. We conclude that some adults deprived of normal binocular vision and insensitive to the disparity information can, with appropriate experience, recover access to more reliable stereoscopic information.This article is part of the themed issue 'Vision in our three-dimensional world'. © 2016 The Author(s).

Recovering stereo vision by squashing virtual bugs in a virtual reality environment

Science.gov (United States)

Vedamurthy, Indu; Knill, David C.; Huang, Samuel J.; Yung, Amanda; Ding, Jian; Kwon, Oh-Sang; Bavelier, Daphne

2016-01-01

Stereopsis is the rich impression of three-dimensionality, based on binocular disparity—the differences between the two retinal images of the same world. However, a substantial proportion of the population is stereo-deficient, and relies mostly on monocular cues to judge the relative depth or distance of objects in the environment. Here we trained adults who were stereo blind or stereo-deficient owing to strabismus and/or amblyopia in a natural visuomotor task—a ‘bug squashing’ game—in a virtual reality environment. The subjects' task was to squash a virtual dichoptic bug on a slanted surface, by hitting it with a physical cylinder they held in their hand. The perceived surface slant was determined by monocular texture and stereoscopic cues, with these cues being either consistent or in conflict, allowing us to track the relative weighting of monocular versus stereoscopic cues as training in the task progressed. Following training most participants showed greater reliance on stereoscopic cues, reduced suppression and improved stereoacuity. Importantly, the training-induced changes in relative stereo weights were significant predictors of the improvements in stereoacuity. We conclude that some adults deprived of normal binocular vision and insensitive to the disparity information can, with appropriate experience, recover access to more reliable stereoscopic information. This article is part of the themed issue ‘Vision in our three-dimensional world’. PMID:27269607

Sequence Analysis of the Capsid Gene during a Genotype II.4 Dominated Norovirus Season in One University Hospital

DEFF Research Database (Denmark)

Holzknecht, Barbara Juliane; Franck, Kristina Træholt; Nielsen, Rikke Thoft

2015-01-01

Norovirus (NoV) is a leading cause of gastroenteritis and genotype II.4 (GII.4) is responsible for the majority of nosocomial NoV infections. Our objective was to examine whether sequencing of the capsid gene might be a useful tool for the hospital outbreak investigation to define possible...

Cell culture adaptation mutations in foot-and-mouth disease virus serotype A capsid proteins: implications for receptor interactions

Science.gov (United States)

In this study we describe the adaptive changes fixed on the capsid of several foot-and-mouth disease virus serotype A strains during propagation in cell monolayers. Viruses passaged extensively in three cell lines (BHK-21, LFBK and IB-RS-2), consistently gained several positively charged amino acids...

Security Vulnerability Profiles of Mission Critical Software: Empirical Analysis of Security Related Bug Reports

Science.gov (United States)

Goseva-Popstojanova, Katerina; Tyo, Jacob

2017-01-01

While some prior research work exists on characteristics of software faults (i.e., bugs) and failures, very little work has been published on analysis of software applications vulnerabilities. This paper aims to contribute towards filling that gap by presenting an empirical investigation of application vulnerabilities. The results are based on data extracted from issue tracking systems of two NASA missions. These data were organized in three datasets: Ground mission IVV issues, Flight mission IVV issues, and Flight mission Developers issues. In each dataset, we identified security related software bugs and classified them in specific vulnerability classes. Then, we created the security vulnerability profiles, i.e., determined where and when the security vulnerabilities were introduced and what were the dominating vulnerabilities classes. Our main findings include: (1) In IVV issues datasets the majority of vulnerabilities were code related and were introduced in the Implementation phase. (2) For all datasets, around 90 of the vulnerabilities were located in two to four subsystems. (3) Out of 21 primary classes, five dominated: Exception Management, Memory Access, Other, Risky Values, and Unused Entities. Together, they contributed from 80 to 90 of vulnerabilities in each dataset.

Competition between the phytophagous stink bugs Euschistus heros and Piezodorus guildinii in soybeans.

Science.gov (United States)

Tuelher, Edmar S; Silva, Éder H; Hirose, Edson; Guedes, Raul Narciso C; Oliveira, Eugênio E

2016-10-01

The abundance and contribution of the neotropical brown stink bug, Euschistus heros (F.), and the redbanded stink bug, Piezodorus guildinii (West.), to the composition of insect pests of soybean, Glycine max (L.), fields have changed both spatially and temporally in neotropical soybean production areas. Therefore, we assessed the competitiveness of each species in direct competition experiments following an additive series. We performed mixed (adult) insect infestations in soybean plants and evaluated the fitness of each species and the soybean yield. While the competitive ability of E. heros was significantly compromised by increments in conspecifics and heterospecifics (i.e. P. guildinii), the competitive ability of P. guildinii was compromised by the presence of heterospecifics (i.e. E. heros). The reproductive output of P. guildinii remained unaffected by increments in E. heros or of P. guildinii. Intriguingly, despite the fact that P. guildinii apparently lost the competition with E. heros, almost no pod production was observed in any plant colonised by the former. The higher abundance of E. heros in neotropical soybean fields seems to result from higher competitive ability than its heterospecific competitor P. guildinii, which may prevent the higher losses caused by P. guildinii. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

Rapid increase of near atomic resolution virus capsid structures determined by cryo-electron microscopy.

Science.gov (United States)

Ho, Phuong T; Reddy, Vijay S

2018-01-01

The recent technological advances in electron microscopes, detectors, as well as image processing and reconstruction software have brought single particle cryo-electron microscopy (cryo-EM) into prominence for determining structures of bio-molecules at near atomic resolution. This has been particularly true for virus capsids, ribosomes, and other large assemblies, which have been the ideal specimens for structural studies by cryo-EM approaches. An analysis of time series metadata of virus structures on the methods of structure determination, resolution of the structures, and size of the virus particles revealed a rapid increase in the virus structures determined by cryo-EM at near atomic resolution since 2010. In addition, the data highlight the median resolution (∼3.0 Å) and size (∼310.0 Å in diameter) of the virus particles determined by X-ray crystallography while no such limits exist for cryo-EM structures, which have a median diameter of 508 Å. Notably, cryo-EM virus structures in the last four years have a median resolution of 3.9 Å. Taken together with minimal sample requirements, not needing diffraction quality crystals, and being able to achieve similar resolutions of the crystal structures makes cryo-EM the method of choice for current and future virus capsid structure determinations. Copyright © 2017 Elsevier Inc. All rights reserved.

From green architecture to architectural green

DEFF Research Database (Denmark)

Earon, Ofri

2011-01-01

that describes the architectural exclusivity of this particular architecture genre. The adjective green expresses architectural qualities differentiating green architecture from none-green architecture. Currently, adding trees and vegetation to the building’s facade is the main architectural characteristics...... they have overshadowed the architectural potential of green architecture. The paper questions how a green space should perform, look like and function. Two examples are chosen to demonstrate thorough integrations between green and space. The examples are public buildings categorized as pavilions. One......The paper investigates the topic of green architecture from an architectural point of view and not an energy point of view. The purpose of the paper is to establish a debate about the architectural language and spatial characteristics of green architecture. In this light, green becomes an adjective...

Structure, Immunogenicity, and Protective Mechanism of an Engineered Enterovirus 71-Like Particle Vaccine Mimicking 80S Empty Capsid.

Science.gov (United States)

Wang, Xiaoli; Ku, Zhiqiang; Zhang, Xiang; Ye, Xiaohua; Chen, Jinhuan; Liu, Qingwei; Zhang, Wei; Zhang, Chao; Fu, Zhenglin; Jin, Xia; Cong, Yao; Huang, Zhong

2018-01-01

Enterovirus 71 (EV71) is the major causative agent of severe hand, foot, and mouth disease, which affects millions of young children in the Asia-Pacific region annually. In this study, we engineered a novel EV71 virus-like particle (VLP) that lacks VP4 (therefore designated VLP ΔVP4 ) and investigated its structure, antigenicity, and vaccine potential. The cryo-electron microscopy (cryo-EM) structure of VLP ΔVP4 was reconstructed to 3.71-Å resolution. Results from structural and biochemical analyses revealed that VLP ΔVP4 resembles the end product of the viral uncoating process, the 80S empty capsid. VLP ΔVP4 is able to elicit high-titer neutralizing antibodies and to fully protect mice against lethal viral challenge. Mechanistic studies showed that, at the cellular level, the anti-VLP ΔVP4 sera exert neutralization effects at both pre- and postattachment stages by inhibiting both virus attachment and internalization, and at the molecular level, the antisera can block multiple interactions between EV71 and its key receptors. Our study gives a better understanding of EV71 capsid assembly and provides important information for the design and development of new-generation vaccines for EV71, and perhaps for other enteroviruses, as well. IMPORTANCE Enterovirus 71 (EV71) infection may lead to severe hand, foot, and mouth disease, with significant morbidity and mortality. Knowledge regarding EV71 particle assembly remains limited. Here, we report the generation and characterization of a novel EV71 virus-like particle that lacks the VP4 capsid subunit protein. This particle, termed VLP ΔVP4 , structurally mimics the 80S empty capsid, which is the end stage of EV71 uncoating. We further show that VLP ΔVP4 exhibits desirable immunogenicity and protective efficacy in proof-of-concept studies. In addition, the inhibitory mechanisms of the VLP ΔVP4 -induced antibodies are unraveled at both the cellular and molecular levels. Our work provides the first evidence of

Detection of fungi from rice black bug Paraeucosmetus pallicornis Dallas (Hemiptera: Lygaeidae) and inhibition with crude extract of Calatropis gigantea (Asclepiadaceae)

Science.gov (United States)

Sjam, S.; Surapati, U.; Adiwena; Syatri, A.; Dewi, V. S.; Rosmana, A.

2018-05-01

Rice black bug (P. palicornis) is one of the pests that attack the rice plants in the generative phase that causes the rice easily destroyed when milled and tasted bitter after cooking hence reduces the quality and quantity of rice. The bitter taste in rice may be due to the fungus associated with rice black bug. The aimed of this research was to detect the associated fungi with rice black bug P. pallicornis using some sterilization methods and inhibition with of leaf crude extract of C. gigatea. Detection of fungi from P. pallicornis was conducted using three sterilization methods and control (without sterilization) namely: (1) sterilization with aquades + alcohol 70% (5, 10, 15 and 20 times dipping) + aquades; (2) aquades + alcohol 70% (10 and 20 times dipping); (3) Aquades + alcohol 90 %+ NaCl 0.5 % +alcohol 90 % + aquades. Inhibition of fungi from P. pallicornis with crude extract of C. gigantea obtained by maceration method and then made some concentration to see the effect of its inhibition on the fungi associated with the P. pallicornis. The results showed that without sterilization, four microbe were obtained: Gliocladium sp., Aspergillus sp., black hyphae fungus and white hyphae fungus, sterilization method of Aquades + alcohol 70% with 5 times dipping in alcohol obtained Gliocladium sp., 10 and 20 times dipping found Aspergillus sp. and Gliocladium sp and 15 times dipping found Aspergillus sp. Sterilization with 10 and 20 times dipping in alcohol 70% then washing 2 times with aquades found Gliogladium sp. and Aspergillus sp. Sterilization with Aquades + alcohol 90 % + NaCl 0.5% + alcohol 90% + aquades found Gliocladium sp. Crude extract of C. gigantea had the potential to inhibit fungi Aspergillus sp. and Gliocladium sp. from rice black bug.

Cyclophilin A interacts with diverse lentiviral capsids

Directory of Open Access Journals (Sweden)

Emerman Michael

2006-10-01

Full Text Available Abstract Background The capsid (CA protein of HIV-1 binds with high affinity to the host protein cyclophilin A (CypA. This binding positively affects some early stage of the viral life-cycle because prevention of binding either by drugs that occupy that active site of cyclophilin A, by mutation in HIV-1 CA, or RNAi that knocks down intracellular CypA level diminishes viral infectivity. The closely related lentivirus, SIVcpz also binds CypA, but it was thought that this interaction was limited to the HIV-1/SIVcpz lineage because other retroviruses failed to interact with CypA in a yeast two-hybrid assay. Results We find that diverse lentiviruses, FIV and SIVagmTAN also bind to CypA. Mutagenesis of FIV CA showed that an amino acid that is in a homologous position to the proline at amino acid 90 of HIV-1 CA is essential for FIV interactions with CypA. Conclusion These results demonstrate that CypA binding to lentiviruses is more widespread than previously thought and suggest that this interaction is evolutionarily important for lentiviral infection.

The Influence of Proactive Green Innovation and Reactive Green Innovation on Green Product Development Performance: The Mediation Role of Green Creativity

Directory of Open Access Journals (Sweden)

Yu-Shan Chen

2016-09-01

Full Text Available This study fills the research gap in the exploration of the relationships between both proactive and reactive green innovations and green product development performance, and examines the mediating effect of green creativity. Structural equation modeling (SEM is utilized to test the hypotheses. From the sample of 146 valid respondents, the results show that proactive green innovation positively affects green creativity and green product development performance, and green creativity positively affects green product development performance. In addition, our findings also indicate that the relationship between proactive green innovation and green product development performance is partially mediated by green creativity. Accordingly, green creativity plays a critical role for companies to achieve a great green product development performance. However, reactive green innovation does not significantly influence green creativity and green product development performance. Companies should develop proactive green innovation rather than reactive green innovation in order to enhance their green creativity and increase their product development performance.

Exposure of bed bugs to metarhizium anisopliae, and the effect of defensive secretions on fungal growth in vitro

Science.gov (United States)

Bed bugs Cimex lectularius were treated with conidia of the entomopathogenic fungus Metarhizium anisopliae by topical, spray, and contact exposure. One week post-exposure, inconsistent mortalities were observed, averaging 30% across all treatment groups and replicates. Microscopic examination of top...

In-vitro replication of Chelonid herpesvirus 5 in organotypic skin cultures from Hawaiian green turtles (Chelonia mydas)

Science.gov (United States)

Work, Thierry M.; Dagenais, Julie; Weatherby, Tina; Ackermann, Mathias; Balazs, George H.

2017-01-01

Fibropapillomatosis (FP) is a tumor disease of marine turtles associated with Chelonid herpesvirus 5 (ChHV5) that has historically been refractory to growth in tissue culture. Here, we show for the first time de novo formation of ChHV5-positive intranuclear inclusions in cultured green turtle cells, which is indicative for active lytic replication of the virus. The minimal requirements to achieve lytic replication in cultured cells included 1) either in-vitro culturing of ChHV5-positive tumor biopsies (plugs) or organotypic cultures (rafts) consisting of ChHV5-positive turtle fibroblasts in collagen rafts seeded with turtle keratinocytes and 2) keratinocyte maturation induced by raising raft or biopsy cultures to the air-liquid interface. Virus growth was confirmed by detailed electron microscopic studies revealing intranuclear sun-shaped capsid factories, tubules, various stages of capsid formation, nuclear export by budding into the perinuclear space, tegumentation, and envelopment to complete de novo virus production. Membrane synthesis was also observed as a sign for active viral replication. Interestingly, cytoplasmic particles became associated with keratin filaments, a feature not seen in conventional monolayer cell cultures where most studies of herpesvirus replication have been performed. Our findings draw a rich and realistic picture of ChHV5 replication in cells derived from its natural host and may be crucial not only to better understand ChHV5 circulation but also to eventually complete Koch's postulates for FP. Moreover, the principles described here may serve as model to culture other viruses that are resistant to replication in conventional cell culture.

Pengaruh Green Marketing Hotel Terhadap Green Consumer Behavior

OpenAIRE

Yo Fernandez, Eunike Christe; Tjoanda, Evelyn

2017-01-01

Penelitian ini dilakukan untuk mengetahui pengaruh dari green marketing hotel terhadap green consumer behavior. Green marketing memiliki 3 dimensi, yaitu green product, green price, dan green promotion. Penelitian ini melibatkan 272 responden masyarakat Surabaya dan menggunakan metode regresi linear berganda. Hasil penelitian menunjukkan bahwa green product dan green price berpengaruh secara positif dan signifikan sedangkan green promotion berpengaruh namun tidak signifikan terhadap green con...

Impact of Bug-in-Ear Professional Development on Early Childhood Co-Teachers' Use of Communication Strategies

Science.gov (United States)

Ottley, Jennifer R.; Grygas Coogle, Christan; Rahn, Naomi L.; Spear, Caitlin F.

2017-01-01

The goal of this study was to build the capacity of early childhood teachers to implement evidence-based strategies. We investigated the efficacy of professional development with bug-in-ear peer coaching in improving teachers' use of communication strategies, the teachers' maintenance of strategies post intervention, and the social validity of the…

Characterization of TES bolometers used in 2-dimensional Backshort-Under-Grid (BUG) arrays for far-infrared astronomy

International Nuclear Information System (INIS)

Staguhn, J.G.; Allen, C.A.; Benford, D.J.; Chervenak, J.A.; Chuss, D.T.; Miller, T.M.; Moseley, S.H.; Wollack, E.J.

2006-01-01

We have produced a laboratory demonstration of our new Backshort-Under-Grid (BUG) bolometer array architecture in a monolithic, 2-dimensional, 8x8 format. The detector array is designed as a square grid of suspended, 1μm thick silicon bolometers with superconducting molybdium/gold bilayer TESs. These detectors use an additional layer of gold bars deposited on top of the bilayer, oriented transverse to the direction of the current flow, for the suppression of excess noise. This detector design has earlier been shown to provide near fundamental noise limited device performance. We present results from performance measurements of witness devices. In particular we demonstrate that the inband excess noise level of the TES detectors is less than 20% above the thermodynamic phonon noise limit and not significantly higher out of band at frequencies that cannot be attenuated by the Nyquist filter. Our 8x8 BUG arrays will be used in the near future for astronomical observations in several (sub-)millimeter instruments

Susceptibility of pest Nezara viridula (Heteroptera: Pentatomidae) and parasitoid Trichopoda pennipes (Diptera: Tachinidae) to selected insecticides.

Science.gov (United States)

Tillman, P Glynn

2006-06-01

Susceptibility of the southern green stink bug, Nezara viridula (L.) (Heteroptera: Pentatomidae), and its endoparasitoid Trichopoda pennipes (F.) (Diptera: Tachinidae) to acetamiprid, cyfluthrin, dicrotophos, indoxacarb, oxamyl, and thiamethoxam was compared in residual and oral toxicity tests. In the residual toxicity test, cyfluthrin, dicrotophos, and oxamyl were highly toxic to N. viridula. Thiamethoxam was moderately toxic to these insects. Each of the four insecticides was highly toxic to T. pennipes after prolonged tarsal contact with dried residues of these chemicals. In the oral toxicity test, where N. viridula fed on food covered with insecticide residues, none of the insecticides were toxic to adults of this stink bug, but acetamiprid, dicrotophos, and thiamethoxam were moderately toxic to the nymphs. In the oral toxicity test, where N. viridula fed on a gel-food containing insecticides, cyfluthrin, dicrotophos, oxamyl, and thiamethoxam were highly toxic to this stink bug. In an oral toxicity test using contaminated sugar water, all of the insecticides were highly toxic to T. pennipes. Because insecticides were as toxic, or more toxic, to T. pennipes than to N. viridula, it is extremely important to conserve this parasitoid by applying these insecticides for control of southern green stink bugs only when the pest reaches economic threshold.

BIOLOGICAL CHARACTERISTICS OF PRUSSIAN CARP (CARASSIUS AURATUS GIBELIO (BLOCH, 1782 COMMERCIAL STOCK OF THE DNIEPER-BUG ESTUARY

Directory of Open Access Journals (Sweden)

К. Heina

2017-09-01

Full Text Available Purpose. To provide the biological assessment of the silver Prussian carp (Carassius auratus gibelio (Bloch, 1782 commercial stock of the Dnieper-Bug estuary in the conditions of the transformed Dnieper river flow. Methodology. During the analysis of the biological state of the Prussian carp commercial stock, the main attention was given to the dynamics of age and sexual structure, length-weight growth rate, absolute fecundity and condition factor. The basic data were collected during the work of control-observation stations of the Institute of Fisheries in the Dnieper-Bug estuary during the current century. The collection and processing of ichthyological materials were performed in accordance with the generally accepted methodologies. Findings. The analysis showed that during the current century, the age structure of the Prussian carp of the Dnieper-Bug estuary was the most labile among other commercial cyprinids. It was found that as a result of an increase in the right wing of the age series, there was a gradual increase of the mean weighted age of its commercial stock. At the beginning of studies (2001-2002, the core of the stock was formed by age-3-6 fish (up ; however in subsequent years, a displacement of dominant groups toward the dominance of age-4-7 fish (more than 80% of the total stock was observed. At the same time, the relative number of age-3 fish (recruits was at a relatively high level – up to 10.6%. The linear growth varied more intensively until the age-5, but it reduced with ageing and did not show high variability. The body weight most variable was in age-4 fish (Cv=9.62%. The noted insignificant deviations in the body weight growth rate of the right wing of the age series was due to stable predominance of females in the stock structure, which were characterized by a variability of the mean weight as a result of different development of gonads. The dynamics of the age-related changes in the condition factor indicated on a

Commercial-biological characteristics of tulka (Clupeonella cultriventris Nordmann, 1840 of the Dnieper-Bug estuary system

Directory of Open Access Journals (Sweden)

K. Geina

2017-12-01

Full Text Available Purpose. To give biological assessment of the commercial stock of tulka Clupeonella cultriventris (Nordmann, 1840 of the Dnieper-Bug estuary system in the conditions of the reduced Dnieper flow. Methodology. Analysis of the biological state of the tulka stock of the Dnieper-Bug estuary system was performed based on the data on the dynamics of its age structure, linear-weight growth and condition factor. Primary materials were obtained from scientific and commercial catches done during the current century. Field and laboratory processing of the ichthyological materials were performed according to generally accepted methods. Findings. Current commercial fishery situation in the Dnieper-Bug estuary system is characterized by reduced intensity of tulka harvest. Currently, major fish harvest is concentrated in the Mykolaiv fishing ground. As a result of continuous underfishing, a gradual growth of the mean weighed age of the tulka stock is observed with the accumulation of maximum groups of the right wing of the age series. At the beginning of the study (2001-2004, when age group was represented by three groups, the nucleus of the population was formed by age-1 and age-2 fish – 95.7%. However, last years, due to an increase in the number of age-4 fish, a reduction in the percentage of modal groups (age-1 and ahe-2 fish to 76.7% was observed. At the same time, the share of age-3 and age-4 fish gradually increased from 15.8% in 2013 to 23.2% in 2017. Against the background of a relatively instable age structure, the linear growth and body weight growth rates did not change significantly. The highest variability of linear parameters was observed in age-1 fish. With a mean body length of 40.55±0.53 mm (lim=36.2-45.2 mm, deviations from the modal value (Cv were 5.10%. With an increase in the age, the linear growth rate decreased and no significant variability was observed. The highest variability of body weight was typical for age-1 and age-2 fish (Cv

Recovery strategies for fluxes affected by the Gill-Solent WindMaster-Pro "w-boost" firmware bug

Science.gov (United States)

Billesbach, D. P.; Chan, S. W.; Biraud, S.; David, C. R.

2017-12-01

In late 2015 and early 2016, work done by the AmeriFlux Tech Team helped to uncover a bug in the Gill WindMaster Pro sonic anemometers used by many researchers for eddy covariance flux measurements. Gill has addressed this issue and has since sent out a notice that the vertical wind speed component (a critical piece of all eddy covariance fluxes) was being erroneously computed and reported. The problem (known as the w-boost bug) resulted in positive (upward) wind speeds being under-reported by 16.6% and negative (downward) wind speeds being under-reported by 28.9%. This has the potential to cause similar underestimates in fluxes obtained from these instruments. While the manufacturer has offered a firmware upgrade to fix this bug, there exist many data sets that have already been affected by it. Researchers who use the affected units have contributed to numerous data archives (AmeriFlux, FluxNet, ICOS, etc.), and third-party scientists have, in turn used these data in many types of research projects. The volume of affected data over such a long period of time makes a complete reprocessing of the raw data sets impractical. To address this, the AmeriFlux Tech Team has endeavored to develop a method of correcting affected fluxes using only the downloadable data sets that are available from these archives. In a previous poster, we reported preliminary results from a pair of Arctic tundra flux towers, and showed that fluxes could be underestimated by 15% to 20%. In this poster, we present results that extend our study to include a forested site in Equatorial Africa. We also have evaluated methods to estimate flux errors without accessing the raw data sets.

Supporting Early Childhood Educators' Use of Embedded Communication Strategies by Providing Feedback via Bug-in-Ear Technology

Science.gov (United States)

Riggie, Jennifer

2013-01-01

The purpose of this study was to examine the relationship between coaching provided with bug-in-ear technology, the frequency of the early childhood educators' use of targeted communication strategies and children's expressive communication. Four multiple-baseline single-case design experiments were completed to evaluate these relationships.…

Green(ing) infrastructure

CSIR Research Space (South Africa)

Van Wyk, Llewellyn V

2014-03-01

Full Text Available the generation of electricity from renewable sources such as wind, water and solar. Grey infrastructure – In the context of storm water management, grey infrastructure can be thought of as the hard, engineered systems to capture and convey runoff..., pumps, and treatment plants.  Green infrastructure reduces energy demand by reducing the need to collect and transport storm water to a suitable discharge location. In addition, green infrastructure such as green roofs, street trees and increased...

Portal protein functions akin to a DNA-sensor that couples genome-packaging to icosahedral capsid maturation

OpenAIRE

Lokareddy, Ravi K.; Sankhala, Rajeshwer S.; Roy, Ankoor; Afonine, Pavel V.; Motwani, Tina; Teschke, Carolyn M.; Parent, Kristin N.; Cingolani, Gino

2017-01-01

Tailed bacteriophages and herpesviruses assemble infectious particles via an empty precursor capsid (or ?procapsid') built by multiple copies of coat and scaffolding protein and by one dodecameric portal protein. Genome packaging triggers rearrangement of the coat protein and release of scaffolding protein, resulting in dramatic procapsid lattice expansion. Here, we provide structural evidence that the portal protein of the bacteriophage P22 exists in two distinct dodecameric conformations: a...

Direct interaction between two viral proteins, the nonstructural protein 2C and the capsid protein VP3, is required for enterovirus morphogenesis.

Directory of Open Access Journals (Sweden)

Ying Liu

2010-08-01

Full Text Available In spite of decades-long studies, the mechanism of morphogenesis of plus-stranded RNA viruses belonging to the genus Enterovirus of Picornaviridae, including poliovirus (PV, is not understood. Numerous attempts to identify an RNA encapsidation signal have failed. Genetic studies, however, have implicated a role of the non-structural protein 2C(ATPase in the formation of poliovirus particles. Here we report a novel mechanism in which protein-protein interaction is sufficient to explain the specificity in PV encapsidation. Making use of a novel "reporter virus", we show that a quasi-infectious chimera consisting of the capsid precursor of C-cluster coxsackie virus 20 (C-CAV20 and the nonstructural proteins of the closely related PV translated and replicated its genome with wild type kinetics, whereas encapsidation was blocked. On blind passages, encapsidation of the chimera was rescued by a single mutation either in capsid protein VP3 of CAV20 or in 2C(ATPase of PV. Whereas each of the single-mutation variants expressed severe proliferation phenotypes, engineering both mutations into the chimera yielded a virus encapsidating with wild type kinetics. Biochemical analyses provided strong evidence for a direct interaction between 2C(ATPase and VP3 of PV and CAV20. Chimeras of other C-CAVs (CAV20/CAV21 or CAV18/CAV20 were blocked in encapsidation (no virus after blind passages but could be rescued if the capsid and 2C(ATPase coding regions originated from the same virus. Our novel mechanism explains the specificity of encapsidation without apparent involvement of an RNA signal by considering that (i genome replication is known to be stringently linked to translation, (ii morphogenesis is known to be stringently linked to genome replication, (iii newly synthesized 2C(ATPase is an essential component of the replication complex, and (iv 2C(ATPase has specific affinity to capsid protein(s. These conditions lead to morphogenesis at the site where newly

Modeling capsid kinetics assembly from the steady state distribution of multi-sizes aggregates

Energy Technology Data Exchange (ETDEWEB)

Hozé, Nathanaël; Holcman, David

2014-01-24

The kinetics of aggregation for particles of various sizes depends on their diffusive arrival and fusion at a specific nucleation site. We present here a mean-field approximation and a stochastic jump model for aggregates at equilibrium. This approach is an alternative to the classical Smoluchowski equations that do not have a close form and are not solvable in general. We analyze these mean-field equations and obtain the kinetics of a cluster formation. Our approach provides a simplified theoretical framework to study the kinetics of viral capsid formation, such as HIV from the self-assembly of the structural proteins Gag.

Maladaptive Plasticity Masks the Effects of Natural Selection in the Red-Shouldered Soapberry Bug.

Science.gov (United States)

Cenzer, Meredith L

2017-10-01

Natural selection can produce local adaptation, but local adaptation can be masked by maladaptive plasticity. Maladaptive plasticity may arise as a result of gene flow producing novel gene combinations that have not been exposed to selection. In the 1980s, populations of the red-shouldered soapberry bug (Jadera haematoloma) were locally adapted to feed on the seeds of a native host plant and an introduced host plant; by 2014, local differentiation in beak length had been lost, likely as a consequence of increased gene flow. In this study, I assess the relative contributions of natural selection and plasticity to beak length on these two hosts. I confirm the earlier hypothesis that the host plant seedpod drives divergent natural selection on beak length. I then demonstrate that the proximate cause of the loss of observable differentiation in beak length is maladaptive plasticity, which masks persistent genetic differences between host-associated populations. Maladaptive plasticity is highest in areas where the two plants co-occur; in combination with historical measures of plasticity in hybrids, this indicates that maladaptive plasticity may be a consequence of ongoing gene flow. Although natural selection produced locally adapted genotypes in soapberry bugs, maladaptive plasticity is masking phenotypic differences between populations in nature.

Molecular markers reveal infestation dynamics of the bed bug (Hemiptera: Cimicidae) within apartment buildings.

Science.gov (United States)

Booth, Warren; Saenz, Virna L; Santangelo, Richard G; Wang, Changlu; Schal, Coby; Vargo, Edward L

2012-05-01

The bed bug, Cimex lectularius L. (Hemiptera: Cimicidae), has experienced an extraordinary global resurgence in recent years, the reasons for which remain poorly understood. Once considered a pest of lower socioeconomic classes, bed bugs are now found extensively across all residential settings, with widespread infestations established in multiapartment buildings. Within such buildings, understanding the population genetic structure and patterns of dispersal may prove critical to the development of effective control strategies. Here, we describe the development of 24 high-resolution microsatellite markers through next generation 454 pyrosequencing and their application to elucidate infestation dynamics within three multistory apartment buildings in the United States. Results reveal contrasting characteristics potentially representative of geographic or locale differences. In Raleigh, NC, an infestation within an apartment building seemed to have started from a single introduction followed by extensive spread. In Jersey City, NJ, two or more introductions followed by spread are evident in two buildings. Populations within single apartments in all buildings were characterized by high levels of relatedness and low levels of diversity, indicative of foundation from small, genetically depauperate propagules. Regardless of the number of unique introductions, genetic data indicate that spread within buildings is extensive, supporting both active and human-mediated dispersal within and between adjacent rooms or apartments spanning multiple floors.