[Dynamic magnetotherapy use in comprehensive treatment phlegmons of maxillofacial region and mandible fractures].

PubMed

Lepilin, A V; Raĭgorodckiĭ, Iu M; Nozdrachev, V G; Erokina, N L

2007-01-01

145 patients (60 with phlegmons of submandibular and submental regions, 85--with fractures of mandible) were observed and treated with the use of moving pulse magnetic field (MPMF) produced by special apparatus (AMO-ATOS-E, , Saratov-city), 60 patients with the same pathology were treated by traditional physical methods and served as control. Use of MPMF led to quicker patient recovery: quicker reduction (if compared with traditional physical methods of treatment) of oedema and soft tissue inflammatory infiltration, quicker relief from inflammatory reaction (according to data of cytokinin profile), improvement of tissue blood supply in the region of fractures in patients with mandible fractures. As the result--we had shortening treatment terms of such patients and complication number reduction.

[Comparison of classical 2D measurement of scoliosis and 3D measurement using vertebral vectors; advantages for prognosis and treatment evaluation].

PubMed

Illés, Tamás

2011-03-01

The EOS system is a new medical imaging device based on low-dose X-rays, gaseous detectors and dedicated software for 3D reconstruction. It was developed by Nobel prizewinner Georges Charpak. A new concept--the vertebral vector--is used to facilitate the interpretation of EOS data, especially in the horizontal plane. We studied 95 cases of idiopathic scoliosis before and after surgery by means of classical methods and using vertebral vectors, in order to compare the accuracy of the two approaches. The vertebral vector permits simultaneous analysis of the scoliotic curvature in the frontal, sagittal and horizontal planes, as precisely as classical methods. The use of the vertebral vector simplifies and facilitates the interpretation of the mass of information provided by EOS. After analyzing the horizontal data, the first goal of corrective intervention would be to reduce the lateral vertebral deviation. The reduction in vertebral rotation seems less important. This is a new element in the therapeutic management of spinal deformations.

78 FR 34556 - Establishment of Class E Airspace; Tobe, CO

Federal Register 2010, 2011, 2012, 2013, 2014

2013-06-10

... facilitate vectoring of Instrument Flight Rules (IFR) aircraft under control of Denver and Albuquerque Air... Albuquerque ARTCC by vectoring aircraft from en route airspace to terminal areas. This action is necessary for...

Enhancements to NASA's Land Atmosphere Near real-time Capability for EOS (LANCE)

NASA Astrophysics Data System (ADS)

Michael, K.; Davies, D. K.; Schmaltz, J. E.; Boller, R. A.; Mauoka, E.; Ye, G.; Vermote, E.; Harrison, S.; Rinsland, P. L.; Protack, S.; Durbin, P. B.; Justice, C. O.

2016-12-01

NASA's Land, Atmosphere Near real-time Capability for EOS (LANCE) supports application users interested in monitoring a wide variety of natural and man-made phenomena. Near Real-Time (NRT) data and imagery from the AIRS, AMSR2, MISR, MLS, MODIS, OMI and VIIRS instruments are available much quicker than routine processing allows. Most data products are available within 3 hours from satellite observation. NRT imagery are generally available 3-5 hours after observation. This article describes LANCE and enhancements made to LANCE over the last year. These enhancements include: the addition of MISR L1 Georeferenced Radiance and L2 Cloud Motion Vector products, AMSR2 Unified L2B Half-Orbit 25 km EASE-Grid Surface Soil Moisture products and VIIRS VIIRS Day/Night Band, Land Surface Reflectance and Corrected Surface reflectance products. In addition, the selection of LANCE NRT imagery that can be interactively viewed through Worldview and the Global Imagery Browse Services (GIBS) has been expanded. LANCE is also working to ingest and process data from OMPS.

DIY series of genetic cassettes useful in construction of versatile vectors specific for Alphaproteobacteria.

PubMed

Dziewit, Lukasz; Adamczuk, Marcin; Szuplewska, Magdalena; Bartosik, Dariusz

2011-08-01

We have developed a DIY (Do It Yourself) series of genetic cassettes, which facilitate construction of novel versatile vectors for Alphaproteobacteria. All the cassettes are based on defined genetic modules derived from three natural plasmids of Paracoccus aminophilus JCM 7686. We have constructed over 50 DIY cassettes, which differ in structure and specific features. All of them are functional in eight strains representing three orders of Alphaproteobacteria: Rhodobacterales, Rhizobiales and Caulobacterales. Besides various replication and stabilization systems, many of the cassettes also contain selective markers appropriate for Alphaproteobacteria (40 cassettes) and genetic modules responsible for mobilization for conjugal transfer (24 cassettes). All the DIY cassettes are bordered by different types of polylinkers, which facilitate vector construction. Using these DIY cassettes, we have created a set of compatible Escherichia coli-Alphaproteobacteria mobilizable shuttle vectors (high or low copy number in E. coli), which will greatly assist the genetic manipulation of Alphaproteobacteria. Copyright © 2011 Elsevier B.V. All rights reserved.

Rapid construction of a Bacterial Artificial Chromosomal (BAC) expression vector using designer DNA fragments.

PubMed

Chen, Chao; Zhao, Xinqing; Jin, Yingyu; Zhao, Zongbao Kent; Suh, Joo-Won

2014-11-01

Bacterial artificial chromosomal (BAC) vectors are increasingly being used in cloning large DNA fragments containing complex biosynthetic pathways to facilitate heterologous production of microbial metabolites for drug development. To express inserted genes using Streptomyces species as the production hosts, an integration expression cassette is required to be inserted into the BAC vector, which includes genetic elements encoding a phage-specific attachment site, an integrase, an origin of transfer, a selection marker and a promoter. Due to the large sizes of DNA inserted into the BAC vectors, it is normally inefficient and time-consuming to assemble these fragments by routine PCR amplifications and restriction-ligations. Here we present a rapid method to insert fragments to construct BAC-based expression vectors. A DNA fragment of about 130 bp was designed, which contains upstream and downstream homologous sequences of both BAC vector and pIB139 plasmid carrying the whole integration expression cassette. In-Fusion cloning was performed using the designer DNA fragment to modify pIB139, followed by λ-RED-mediated recombination to obtain the BAC-based expression vector. We demonstrated the effectiveness of this method by rapid construction of a BAC-based expression vector with an insert of about 120 kb that contains the entire gene cluster for biosynthesis of immunosuppressant FK506. The empty BAC-based expression vector constructed in this study can be conveniently used for construction of BAC libraries using either microbial pure culture or environmental DNA, and the selected BAC clones can be directly used for heterologous expression. Alternatively, if a BAC library has already been constructed using a commercial BAC vector, the selected BAC vectors can be manipulated using the method described here to get the BAC-based expression vectors with desired gene clusters for heterologous expression. The rapid construction of a BAC-based expression vector facilitates heterologous expression of large gene clusters for drug discovery. Copyright © 2014 Elsevier Inc. All rights reserved.

Visual comparison testing of automotive paint simulation

NASA Astrophysics Data System (ADS)

Meyer, Gary; Fan, Hua-Tzu; Seubert, Christopher; Evey, Curtis; Meseth, Jan; Schnackenberg, Ryan

2015-03-01

An experiment was performed to determine whether typical industrial automotive color paint comparisons made using real physical samples could also be carried out using a digital simulation displayed on a calibrated color television monitor. A special light booth, designed to facilitate evaluation of the car paint color with reflectance angle, was employed in both the real and virtual color comparisons. Paint samples were measured using a multi-angle spectrophotometer and were simulated using a commercially available software package. Subjects performed the test quicker using the computer graphic simulation, and results indicate that there is only a small difference between the decisions made using the light booth and the computer monitor. This outcome demonstrates the potential of employing simulations to replace some of the time consuming work with real physical samples that still characterizes material appearance work in industry.

Current Laser Resurfacing Technologies: A Review that Delves Beneath the Surface

PubMed Central

Preissig, Jason; Hamilton, Kristy; Markus, Ramsey

2012-01-01

Numerous laser platforms exist that rejuvenate the skin by resurfacing its upper layers. In varying degrees, these lasers improve the appearance of lentigines and rhytides, eliminate photoaging, soften scarring due to acne and other causes, and treat dyspigmentation. Five major classes of dermatologic lasers are currently in common use: ablative and nonablative lasers in both fractionated and unfractionated forms as well as radiofrequency technologies. The gentler nonablative lasers allow for quicker healing, whereas harsher ablative lasers tend to be more effective. Fractionating either laser distributes the effect, increasing the number of treatments but minimizing downtime and complications. In this review article, the authors seek to inform surgeons about the current laser platforms available, clarify the differences between them, and thereby facilitate the identification of the most appropriate laser for their practice. PMID:23904818

Regulated Apoptosis of Genetically-Modified Hematopoietic Stem and Progenitor Cells via an Inducible Caspase-9 Suicide Gene in Rhesus Macaques

PubMed Central

Barese, Cecilia N.; Felizardo, Tania C.; Sellers, Stephanie E.; Keyvanfar, Keyvan; Di Stasi, Antonio; Metzger, Mark E.; Krouse, Allen E.; Donahue, Robert E.; Spencer, David M.; Dunbar, Cynthia E.

2014-01-01

The high risk of insertional oncogenesis reported in clinical trials utilizing integrating retroviral vectors to genetically-modify hematopoietic stem and progenitor cells (HSPC) requires the development of safety strategies to minimize risks associated with novel cell and gene therapies. The ability to ablate genetically modified cells in vivo is desirable, should an abnormal clone emerge. Inclusion of “suicide genes” in vectors to facilitate targeted ablation of vector-containing abnormal clones in vivo is one potential safety approach. We tested whether the inclusion of the “inducible Caspase-9” (iCasp9) suicide gene in a gamma-retroviral vector facilitated efficient elimination of vector-containing HSPCs and their hematopoietic progeny in vivo long-term, in an autologous non-human primate transplantation model. Following stable engraftment of iCasp9 expressing hematopoietic cells in rhesus macaques, administration of AP1903, a chemical inducer of dimerization able to activate iCasp9, specifically eliminated vector-containing cells in all hematopoietic lineages long-term, suggesting activity at the HSPC level. Between 75–94% of vector-containing cells were eliminated by well-tolerated AP1903 dosing, but lack of complete ablation was linked to lower iCasp9 expression in residual cells. Further investigation of resistance mechanisms demonstrated upregulation of Bcl-2 in hematopoietic cell lines transduced with the vector and resistant to AP1903 ablation. These results demonstrate both the potential and the limitations of safety approaches utilizing iCasp9 to HSPC-targeted gene therapy settings, in a model with great relevance to clinical development. PMID:25330775

Lentiviral gene ontology (LeGO) vectors equipped with novel drug-selectable fluorescent proteins: new building blocks for cell marking and multi-gene analysis.

PubMed

Weber, K; Mock, U; Petrowitz, B; Bartsch, U; Fehse, B

2010-04-01

Vector-encoded fluorescent proteins (FPs) facilitate unambiguous identification or sorting of gene-modified cells by fluorescence-activated cell sorting (FACS). Exploiting this feature, we have recently developed lentiviral gene ontology (LeGO) vectors (www.LentiGO-Vectors.de) for multi-gene analysis in different target cells. In this study, we extend the LeGO principle by introducing 10 different drug-selectable FPs created by fusing one of the five selection marker (protecting against blasticidin, hygromycin, neomycin, puromycin and zeocin) and one of the five FP genes (Cerulean, eGFP, Venus, dTomato and mCherry). All tested fusion proteins allowed both fluorescence-mediated detection and drug-mediated selection of LeGO-transduced cells. Newly generated codon-optimized hygromycin- and neomycin-resistance genes showed improved expression as compared with their ancestors. New LeGO constructs were produced at titers >10(6) per ml (for non-concentrated supernatants). We show efficient combinatorial marking and selection of various cells, including mesenchymal stem cells, simultaneously transduced with different LeGO constructs. Inclusion of the cytomegalovirus early enhancer/chicken beta-actin promoter into LeGO vectors facilitated robust transgene expression in and selection of neural stem cells and their differentiated progeny. We suppose that the new drug-selectable markers combining advantages of FACS and drug selection are well suited for numerous applications and vector systems. Their inclusion into LeGO vectors opens new possibilities for (stem) cell tracking and functional multi-gene analysis.

TA-GC cloning: A new simple and versatile technique for the directional cloning of PCR products for recombinant protein expression.

PubMed

Niarchos, Athanasios; Siora, Anastasia; Konstantinou, Evangelia; Kalampoki, Vasiliki; Lagoumintzis, George; Poulas, Konstantinos

2017-01-01

During the last few decades, the recombinant protein expression finds more and more applications. The cloning of protein-coding genes into expression vectors is required to be directional for proper expression, and versatile in order to facilitate gene insertion in multiple different vectors for expression tests. In this study, the TA-GC cloning method is proposed, as a new, simple and efficient method for the directional cloning of protein-coding genes in expression vectors. The presented method features several advantages over existing methods, which tend to be relatively more labour intensive, inflexible or expensive. The proposed method relies on the complementarity between single A- and G-overhangs of the protein-coding gene, obtained after a short incubation with T4 DNA polymerase, and T and C overhangs of the novel vector pET-BccI, created after digestion with the restriction endonuclease BccI. The novel protein-expression vector pET-BccI also facilitates the screening of transformed colonies for recombinant transformants. Evaluation experiments of the proposed TA-GC cloning method showed that 81% of the transformed colonies contained recombinant pET-BccI plasmids, and 98% of the recombinant colonies expressed the desired protein. This demonstrates that TA-GC cloning could be a valuable method for cloning protein-coding genes in expression vectors.

TA-GC cloning: A new simple and versatile technique for the directional cloning of PCR products for recombinant protein expression

PubMed Central

Niarchos, Athanasios; Siora, Anastasia; Konstantinou, Evangelia; Kalampoki, Vasiliki; Poulas, Konstantinos

2017-01-01

During the last few decades, the recombinant protein expression finds more and more applications. The cloning of protein-coding genes into expression vectors is required to be directional for proper expression, and versatile in order to facilitate gene insertion in multiple different vectors for expression tests. In this study, the TA-GC cloning method is proposed, as a new, simple and efficient method for the directional cloning of protein-coding genes in expression vectors. The presented method features several advantages over existing methods, which tend to be relatively more labour intensive, inflexible or expensive. The proposed method relies on the complementarity between single A- and G-overhangs of the protein-coding gene, obtained after a short incubation with T4 DNA polymerase, and T and C overhangs of the novel vector pET-BccI, created after digestion with the restriction endonuclease BccI. The novel protein-expression vector pET-BccI also facilitates the screening of transformed colonies for recombinant transformants. Evaluation experiments of the proposed TA-GC cloning method showed that 81% of the transformed colonies contained recombinant pET-BccI plasmids, and 98% of the recombinant colonies expressed the desired protein. This demonstrates that TA-GC cloning could be a valuable method for cloning protein-coding genes in expression vectors. PMID:29091919

Updating in working memory predicts greater emotion reactivity to and facilitated recovery from negative emotion-eliciting stimuli

PubMed Central

Pe, Madeline L.; Koval, Peter; Houben, Marlies; Erbas, Yasemin; Champagne, Dominique; Kuppens, Peter

2015-01-01

That emotions change in response to emotion-eliciting events is a natural part of human life. However, it is equally important for emotions to return to baseline once the emotion-eliciting events have passed. This suggests that the ability to emotionally react to and recover from emotion-eliciting events is critical for healthy psychological functioning. But why do individuals differ in their emotion reactivity and recovery? The present work postulates that the ability to update emotional information in working memory (WM) may explain individual differences in emotion reactivity and recovery. Two studies are presented, which examined whether updating ability was related to emotion reactivity and recovery. In Study 1, we assessed participants' self-reported affect as they viewed negative and positive films. Our results revealed that better updating ability was related to greater emotion reactivity and facilitated (i.e., quicker) recovery from watching negative films. In Study 2, participants recalled a recent angering event, and were then instructed to either ruminate about or reappraise the event. Results revealed that updating ability was again related to greater emotion reactivity and facilitated (i.e., successful) emotion recovery in response to the angering event, and that this was unrelated to the emotion regulation strategy used. These findings identify the ability to update emotional information in WM as a possible mechanism in emotion responding. PMID:25914655

TMV-Gate vectors: Gateway compatible tobacco mosaic virus based expression vectors for functional analysis of proteins

PubMed Central

Kagale, Sateesh; Uzuhashi, Shihomi; Wigness, Merek; Bender, Tricia; Yang, Wen; Borhan, M. Hossein; Rozwadowski, Kevin

2012-01-01

Plant viral expression vectors are advantageous for high-throughput functional characterization studies of genes due to their capability for rapid, high-level transient expression of proteins. We have constructed a series of tobacco mosaic virus (TMV) based vectors that are compatible with Gateway technology to enable rapid assembly of expression constructs and exploitation of ORFeome collections. In addition to the potential of producing recombinant protein at grams per kilogram FW of leaf tissue, these vectors facilitate either N- or C-terminal fusions to a broad series of epitope tag(s) and fluorescent proteins. We demonstrate the utility of these vectors in affinity purification, immunodetection and subcellular localisation studies. We also apply the vectors to characterize protein-protein interactions and demonstrate their utility in screening plant pathogen effectors. Given its broad utility in defining protein properties, this vector series will serve as a useful resource to expedite gene characterization efforts. PMID:23166857

Manifolds for pose tracking from monocular video

NASA Astrophysics Data System (ADS)

Basu, Saurav; Poulin, Joshua; Acton, Scott T.

2015-03-01

We formulate a simple human-pose tracking theory from monocular video based on the fundamental relationship between changes in pose and image motion vectors. We investigate the natural embedding of the low-dimensional body pose space into a high-dimensional space of body configurations that behaves locally in a linear manner. The embedded manifold facilitates the decomposition of the image motion vectors into basis motion vector fields of the tangent space to the manifold. This approach benefits from the style invariance of image motion flow vectors, and experiments to validate the fundamental theory show reasonable accuracy (within 4.9 deg of the ground truth).

Support vector machine applied to predict the zoonotic potential of E. coli O157 cattle isolates

USDA-ARS?s Scientific Manuscript database

Methods based on sequence data analysis facilitate the tracking of disease outbreaks, allow relationships between strains to be reconstructed and virulence factors to be identified. However, these methods are used postfactum after an outbreak has happened. Here, we show that support vector machine a...

Item Vector Plots for the Multidimensional Three-Parameter Logistic Model

ERIC Educational Resources Information Center

Bryant, Damon; Davis, Larry

2011-01-01

This brief technical note describes how to construct item vector plots for dichotomously scored items fitting the multidimensional three-parameter logistic model (M3PLM). As multidimensional item response theory (MIRT) shows promise of being a very useful framework in the test development life cycle, graphical tools that facilitate understanding…

A modular toolset for recombination transgenesis and neurogenetic analysis of Drosophila.

PubMed

Wang, Ji-Wu; Beck, Erin S; McCabe, Brian D

2012-01-01

Transgenic Drosophila have contributed extensively to our understanding of nervous system development, physiology and behavior in addition to being valuable models of human neurological disease. Here, we have generated a novel series of modular transgenic vectors designed to optimize and accelerate the production and analysis of transgenes in Drosophila. We constructed a novel vector backbone, pBID, that allows both phiC31 targeted transgene integration and incorporates insulator sequences to ensure specific and uniform transgene expression. Upon this framework, we have built a series of constructs that are either backwards compatible with existing restriction enzyme based vectors or utilize Gateway recombination technology for high-throughput cloning. These vectors allow for endogenous promoter or Gal4 targeted expression of transgenic proteins with or without fluorescent protein or epitope tags. In addition, we have generated constructs that facilitate transgenic splice isoform specific RNA inhibition of gene expression. We demonstrate the utility of these constructs to analyze proteins involved in nervous system development, physiology and neurodegenerative disease. We expect that these reagents will facilitate the proficiency and sophistication of Drosophila genetic analysis in both the nervous system and other tissues.

Developmental neurogenetics of sexual dimorphism in Aedes aegypti

PubMed Central

Duman-Scheel, Molly; Syed, Zainulabeuddin

2015-01-01

Sexual dimorphism, a poorly understood but crucial aspect of vector mosquito biology, encompasses sex-specific physical, physiological, and behavioral traits related to mosquito reproduction. The study of mosquito sexual dimorphism has largely focused on analysis of the differences between adult female and male mosquitoes, particularly with respect to sex-specific behaviors related to disease transmission. However, sexually dimorphic behaviors are the products of differential gene expression that initiates during development and therefore must also be studied during development. Recent technical advancements are facilitating functional genetic studies in the dengue vector Aedes aegypti, an emerging model for mosquito development. These methodologies, many of which could be extended to other non-model insect species, are facilitating analysis of the development of sexual dimorphism in neural tissues, particularly the olfactory system. These studies are providing insight into the neurodevelopmental genetic basis for sexual dimorphism in vector mosquitoes. PMID:26949699

A new lifetime estimation model for a quicker LED reliability prediction

NASA Astrophysics Data System (ADS)

Hamon, B. H.; Mendizabal, L.; Feuillet, G.; Gasse, A.; Bataillou, B.

2014-09-01

LED reliability and lifetime prediction is a key point for Solid State Lighting adoption. For this purpose, one hundred and fifty LEDs have been aged for a reliability analysis. LEDs have been grouped following nine current-temperature stress conditions. Stress driving current was fixed between 350mA and 1A and ambient temperature between 85C and 120°C. Using integrating sphere and I(V) measurements, a cross study of the evolution of electrical and optical characteristics has been done. Results show two main failure mechanisms regarding lumen maintenance. The first one is the typically observed lumen depreciation and the second one is a much more quicker depreciation related to an increase of the leakage and non radiative currents. Models of the typical lumen depreciation and leakage resistance depreciation have been made using electrical and optical measurements during the aging tests. The combination of those models allows a new method toward a quicker LED lifetime prediction. These two models have been used for lifetime predictions for LEDs.

Short-Term Safety and Efficacy of Femoral Vascular Closure after Percutaneous Coronary Intervention with Combination of the Boomerang(TM) Device and Intravenous Protamine Sulfate.

PubMed

Chen, Ching-Pei; Huang, Huang-Kai; Hsia, Chien-Hsun; Chang, Yung-Ming; Lin, Lee-Shin; Lee, Cheng-Liang

2013-11-01

The Cardiva Boomerang(TM) is a device used to perform femoral vascular closure. It facilitates passive hemostasis at the arteriotomy site, leaving no residual foreign body. We performed a controlled, randomized study of 60 patients undergoing percutaneous coronary intervention. Patients were randomized into two groups (30 per group) to undergo vascular closure with the Boomerang(TM) or the Perclose(TM) suture-based device after the intravenous administration of protamine sulfate. We compared overall success rates, patient-reported pain, length of time to achieve hemostasis and mobilization of the patient, and the frequency of complications in the two groups. Overall success rates using the Boomerang(TM) and Perclose(TM) devices were similarly high, at 93% and 97%, respectively. The Boomerang(TM) was significantly quicker to deploy than the PercloseTM, device deployment time, median (Q1-Q3), [2.00 (1.33-2.75) vs. 3.84 (2.75-4.38) mins, p < 0.001)]. The pain score was significantly lower in the Boomerang(TM) group (1.1 ± 1.7 vs. 6.4 ± 2.9, p < 0.001). The time the device remained in the artery and manual compression time were significantly longer with the Boomerang(TM) (p < 0.001), as well as the time taken to achieve hemostasis and time to ambulation. There were no major complications in either group and no significant differences between the groups in the frequency of minor complications. We conclude that when used in combination with intravenous protamine sulfate, the Boomerang(TM) device is as safe and effective as the Perclose(TM) device for femoral vascular closure, but quicker to deploy and less painful to patients. Boomerang; Percutaneous intervention; Vascular closure device.

Usability Evaluation of an Augmented Reality System for Teaching Euclidean Vectors

ERIC Educational Resources Information Center

Martin-Gonzalez, Anabel; Chi-Poot, Angel; Uc-Cetina, Victor

2016-01-01

Augmented reality (AR) is one of the emerging technologies that has demonstrated to be an efficient technological tool to enhance learning techniques. In this paper, we describe the development and evaluation of an AR system for teaching Euclidean vectors in physics and mathematics. The goal of this pedagogical tool is to facilitate user's…

VectorBase: an updated bioinformatics resource for invertebrate vectors and other organisms related with human diseases

PubMed Central

Giraldo-Calderón, Gloria I.; Emrich, Scott J.; MacCallum, Robert M.; Maslen, Gareth; Dialynas, Emmanuel; Topalis, Pantelis; Ho, Nicholas; Gesing, Sandra; Madey, Gregory; Collins, Frank H.; Lawson, Daniel

2015-01-01

VectorBase is a National Institute of Allergy and Infectious Diseases supported Bioinformatics Resource Center (BRC) for invertebrate vectors of human pathogens. Now in its 11th year, VectorBase currently hosts the genomes of 35 organisms including a number of non-vectors for comparative analysis. Hosted data range from genome assemblies with annotated gene features, transcript and protein expression data to population genetics including variation and insecticide-resistance phenotypes. Here we describe improvements to our resource and the set of tools available for interrogating and accessing BRC data including the integration of Web Apollo to facilitate community annotation and providing Galaxy to support user-based workflows. VectorBase also actively supports our community through hands-on workshops and online tutorials. All information and data are freely available from our website at https://www.vectorbase.org/. PMID:25510499

Computer-assisted propofol administration.

PubMed

O'Connor, J P A; O'Moráin, C A; Vargo, J J

2010-01-01

The use of propofol for sedation in endoscopy may allow for better quality of sedation, quicker recovery and facilitate greater throughput in endoscopy units. The cost-effectiveness and utility of propofol sedation for endoscopic procedures is contingent on the personnel and resources required to carry out the procedure. Computer-based platforms are based on the patients response to stimulation and physiologic parameters. They offer an appealing means of delivering safe and effective doses of propofol. One such means is the bispectral index where continuous EEG recordings are used to assess the degree of sedation. Another is the closed-loop target-controlled system where a set of physical parameters, such as muscle relaxation and auditory-evoked potential, determine a level of medication appropriate to achieve sedation. Patient-controlled platforms may also be used. These electronic adjuncts may help endoscopists who wish to adopt propofol sedation to change current practices with greater confidence. Copyright 2010 S. Karger AG, Basel.

Vaxvec: The first web-based recombinant vaccine vector database and its data analysis

PubMed Central

Deng, Shunzhou; Martin, Carly; Patil, Rasika; Zhu, Felix; Zhao, Bin; Xiang, Zuoshuang; He, Yongqun

2015-01-01

A recombinant vector vaccine uses an attenuated virus, bacterium, or parasite as the carrier to express a heterologous antigen(s). Many recombinant vaccine vectors and related vaccines have been developed and extensively investigated. To compare and better understand recombinant vectors and vaccines, we have generated Vaxvec (http://www.violinet.org/vaxvec), the first web-based database that stores various recombinant vaccine vectors and those experimentally verified vaccines that use these vectors. Vaxvec has now included 59 vaccine vectors that have been used in 196 recombinant vector vaccines against 66 pathogens and cancers. These vectors are classified to 41 viral vectors, 15 bacterial vectors, 1 parasitic vector, and 1 fungal vector. The most commonly used viral vaccine vectors are double-stranded DNA viruses, including herpesviruses, adenoviruses, and poxviruses. For example, Vaxvec includes 63 poxvirus-based recombinant vaccines for over 20 pathogens and cancers. Vaxvec collects 30 recombinant vector influenza vaccines that use 17 recombinant vectors and were experimentally tested in 7 animal models. In addition, over 60 protective antigens used in recombinant vector vaccines are annotated and analyzed. User-friendly web-interfaces are available for querying various data in Vaxvec. To support data exchange, the information of vaccine vectors, vaccines, and related information is stored in the Vaccine Ontology (VO). Vaxvec is a timely and vital source of vaccine vector database and facilitates efficient vaccine vector research and development. PMID:26403370

Quick fuzzy backpropagation algorithm.

PubMed

Nikov, A; Stoeva, S

2001-03-01

A modification of the fuzzy backpropagation (FBP) algorithm called QuickFBP algorithm is proposed, where the computation of the net function is significantly quicker. It is proved that the FBP algorithm is of exponential time complexity, while the QuickFBP algorithm is of polynomial time complexity. Convergence conditions of the QuickFBP, resp. the FBP algorithm are defined and proved for: (1) single output neural networks in case of training patterns with different targets; and (2) multiple output neural networks in case of training patterns with equivalued target vector. They support the automation of the weights training process (quasi-unsupervised learning) establishing the target value(s) depending on the network's input values. In these cases the simulation results confirm the convergence of both algorithms. An example with a large-sized neural network illustrates the significantly greater training speed of the QuickFBP rather than the FBP algorithm. The adaptation of an interactive web system to users on the basis of the QuickFBP algorithm is presented. Since the QuickFBP algorithm ensures quasi-unsupervised learning, this implies its broad applicability in areas of adaptive and adaptable interactive systems, data mining, etc. applications.

Large-scale adenovirus and poxvirus-vectored vaccine manufacturing to enable clinical trials.

PubMed

Kallel, Héla; Kamen, Amine A

2015-05-01

Efforts to make vaccines against infectious diseases and immunotherapies for cancer have evolved to utilize a variety of heterologous expression systems such as viral vectors. These vectors are often attenuated or engineered to safely deliver genes encoding antigens of different pathogens. Adenovirus and poxvirus vectors are among the viral vectors that are most frequently used to develop prophylactic vaccines against infectious diseases as well as therapeutic cancer vaccines. This mini-review describes the trends and processes in large-scale production of adenovirus and poxvirus vectors to meet the needs of clinical applications. We briefly describe the general principles for the production and purification of adenovirus and poxvirus viral vectors. Currently, adenovirus and poxvirus vector manufacturing methods rely on well-established cell culture technologies. Several improvements have been evaluated to increase the yield and to reduce the overall manufacturing cost, such as cultivation at high cell densities and continuous downstream processing. Additionally, advancements in vector characterization will greatly facilitate the development of novel vectored vaccine candidates. Copyright © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

A Modular Toolset for Recombination Transgenesis and Neurogenetic Analysis of Drosophila

PubMed Central

Wang, Ji-Wu; Beck, Erin S.; McCabe, Brian D.

2012-01-01

Transgenic Drosophila have contributed extensively to our understanding of nervous system development, physiology and behavior in addition to being valuable models of human neurological disease. Here, we have generated a novel series of modular transgenic vectors designed to optimize and accelerate the production and analysis of transgenes in Drosophila. We constructed a novel vector backbone, pBID, that allows both phiC31 targeted transgene integration and incorporates insulator sequences to ensure specific and uniform transgene expression. Upon this framework, we have built a series of constructs that are either backwards compatible with existing restriction enzyme based vectors or utilize Gateway recombination technology for high-throughput cloning. These vectors allow for endogenous promoter or Gal4 targeted expression of transgenic proteins with or without fluorescent protein or epitope tags. In addition, we have generated constructs that facilitate transgenic splice isoform specific RNA inhibition of gene expression. We demonstrate the utility of these constructs to analyze proteins involved in nervous system development, physiology and neurodegenerative disease. We expect that these reagents will facilitate the proficiency and sophistication of Drosophila genetic analysis in both the nervous system and other tissues. PMID:22848718

New ΦBT1 site-specific integrative vectors with neutral phenotype in Streptomyces.

PubMed

Gonzalez-Quiñonez, Nathaly; López-García, María Teresa; Yagüe, Paula; Rioseras, Beatriz; Pisciotta, Annalisa; Alduina, Rosa; Manteca, Ángel

2016-03-01

Integrative plasmids are one of the best options to introduce genes in low copy and in a stable form into bacteria. The ΦC31-derived plasmids constitute the most common integrative vectors used in Streptomyces. They integrate at different positions (attB and pseudo-attB sites) generating different mutations. The less common ΦBT1-derived vectors integrate at the unique attB site localized in the SCO4848 gene (S. coelicolor genome) or their orthologues in other streptomycetes. This work demonstrates that disruption of SCO4848 generates a delay in spore germination. SCO4848 is co-transcribed with SCO4849, and the spore germination phenotype is complemented by SCO4849. Plasmids pNG1-4 were created by modifying the ΦBT1 integrative vector pMS82 by introducing a copy of SCO4849 under the control of the promoter region of SCO4848. pNG2 and pNG4 also included a copy of the P ermE * in order to facilitate gene overexpression. pNG3 and pNG4 harboured a copy of the bla gene (ampicillin resistance) to facilitate selection in E. coli. pNG1-4 are the only integrative vectors designed to produce a neutral phenotype when they are integrated into the Streptomyces genome. The experimental approach developed in this work can be applied to create phenotypically neutral integrative plasmids in other bacteria.

Single Vector Calibration System for Multi-Axis Load Cells and Method for Calibrating a Multi-Axis Load Cell

NASA Technical Reports Server (NTRS)

Parker, Peter A. (Inventor)

2003-01-01

A single vector calibration system is provided which facilitates the calibration of multi-axis load cells, including wind tunnel force balances. The single vector system provides the capability to calibrate a multi-axis load cell using a single directional load, for example loading solely in the gravitational direction. The system manipulates the load cell in three-dimensional space, while keeping the uni-directional calibration load aligned. The use of a single vector calibration load reduces the set-up time for the multi-axis load combinations needed to generate a complete calibration mathematical model. The system also reduces load application inaccuracies caused by the conventional requirement to generate multiple force vectors. The simplicity of the system reduces calibration time and cost, while simultaneously increasing calibration accuracy.

Harnessing Integrated Vector Management for Enhanced Disease Prevention.

PubMed

Chanda, Emmanuel; Ameneshewa, Birkinesh; Bagayoko, Magaran; Govere, John M; Macdonald, Michael B

2017-01-01

The increasing global threat of emerging and re-emerging vector-borne diseases (VBDs) poses a serious health problem. The World Health Organization (WHO) recommends integrated vector management (IVM) strategy for combating VBD transmission. An IVM approach requires entomological knowledge, technical and infrastructure capacity, and systems facilitating stakeholder collaboration. In sub-Saharan Africa, successful operational IVM experience comes from relatively few countries. This article provides an update on the extent to which IVM is official national policy, the degree of IVM implementation, the level of compliance with WHO guidelines, and concordance in the understanding of IVM, and it assesses the operational impact of IVM. The future outlook encompasses rational and sustainable use of effective vector control tools and inherent improved return for investment for disease vector control. Copyright © 2016 Elsevier Ltd. All rights reserved.

Vaxvec: The first web-based recombinant vaccine vector database and its data analysis.

PubMed

Deng, Shunzhou; Martin, Carly; Patil, Rasika; Zhu, Felix; Zhao, Bin; Xiang, Zuoshuang; He, Yongqun

2015-11-27

A recombinant vector vaccine uses an attenuated virus, bacterium, or parasite as the carrier to express a heterologous antigen(s). Many recombinant vaccine vectors and related vaccines have been developed and extensively investigated. To compare and better understand recombinant vectors and vaccines, we have generated Vaxvec (http://www.violinet.org/vaxvec), the first web-based database that stores various recombinant vaccine vectors and those experimentally verified vaccines that use these vectors. Vaxvec has now included 59 vaccine vectors that have been used in 196 recombinant vector vaccines against 66 pathogens and cancers. These vectors are classified to 41 viral vectors, 15 bacterial vectors, 1 parasitic vector, and 1 fungal vector. The most commonly used viral vaccine vectors are double-stranded DNA viruses, including herpesviruses, adenoviruses, and poxviruses. For example, Vaxvec includes 63 poxvirus-based recombinant vaccines for over 20 pathogens and cancers. Vaxvec collects 30 recombinant vector influenza vaccines that use 17 recombinant vectors and were experimentally tested in 7 animal models. In addition, over 60 protective antigens used in recombinant vector vaccines are annotated and analyzed. User-friendly web-interfaces are available for querying various data in Vaxvec. To support data exchange, the information of vaccine vectors, vaccines, and related information is stored in the Vaccine Ontology (VO). Vaxvec is a timely and vital source of vaccine vector database and facilitates efficient vaccine vector research and development. Copyright © 2015 Elsevier Ltd. All rights reserved.

Engineering T7 bacteriophage as a potential DNA vaccine targeting delivery vector.

PubMed

Xu, Hai; Bao, Xi; Wang, Yiwei; Xu, Yue; Deng, Bihua; Lu, Yu; Hou, Jibo

2018-03-20

DNA delivery with bacteriophage by surface-displayed mammalian cell penetrating peptides has been reported. Although, various phages have been used to facilitate DNA transfer by surface displaying the protein transduction domain of human immunodeficiency virus type 1 Tat protein (Tat peptide), no similar study has been conducted using T7 phage. In this study, we engineeredT7 phage as a DNA targeting delivery vector to facilitate cellular internalization. We constructed recombinant T7 phages that displayed Tat peptide on their surface and carried eukaryotic expression box (EEB) as a part of their genomes (T7-EEB-Tat). We demonstrated that T7 phage harboring foreign gene insertion had packaged into infective progeny phage particles. Moreover, when mammalian cells that were briefly exposed to T7-EEB-Tat, expressed a significant higher level of the marker gene with the control cells infected with the wide type phage without displaying Tat peptides. These data suggested that the potential of T7 phage as an effective delivery vector for DNA vaccine transfer.

A new image representation for compact and secure communication

DOE Office of Scientific and Technical Information (OSTI.GOV)

Prasad, Lakshman; Skourikhine, A. N.

In many areas of nuclear materials management there is a need for communication, archival, and retrieval of annotated image data between heterogeneous platforms and devices to effectively implement safety, security, and safeguards of nuclear materials. Current image formats such as JPEG are not ideally suited in such scenarios as they are not scalable to different viewing formats, and do not provide a high-level representation of images that facilitate automatic object/change detection or annotation. The new Scalable Vector Graphics (SVG) open standard for representing graphical information, recommended by the World Wide Web Consortium (W3C) is designed to address issues of imagemore » scalability, portability, and annotation. However, until now there has been no viable technology to efficiently field images of high visual quality under this standard. Recently, LANL has developed a vectorized image representation that is compatible with the SVG standard and preserves visual quality. This is based on a new geometric framework for characterizing complex features in real-world imagery that incorporates perceptual principles of processing visual information known from cognitive psychology and vision science, to obtain a polygonal image representation of high fidelity. This representation can take advantage of all textual compression and encryption routines unavailable to other image formats. Moreover, this vectorized image representation can be exploited to facilitate automated object recognition that can reduce time required for data review. The objects/features of interest in these vectorized images can be annotated via animated graphics to facilitate quick and easy display and comprehension of processed image content.« less

Strengthening and expanding the capacity of health worker education in Zambia

PubMed Central

Michelo, Charles; Zulu, Joseph Mumba; Simuyemba, Moses; Andrews, Benjamin; Katubulushi, Max; Chi, Benjamin; Njelesani, Evariste; Vwalika, Bellington; Bowa, Kasonde; Maimbolwa, Margaret; Chipeta, James; Goma, Fastone; Nzala, Selestine; Banda, Sekelani; Mudenda, John; Ahmed, Yusuf; Hachambwa, Lotti; Wilson, Craig; Vermund, Sten; Mulla, Yakub

2017-01-01

Introduction Zambia is facing a chronic shortage of health care workers. The paper aimed at understanding how the Medical Education Partnership Initiative (MEPI) program facilitated strengthening and expanding of the national capacity and quality of medical education as well as processes for retaining faculty in Zambia. Methods Data generated through documentary review, key informant interviews and observations were analyzed using a thematic approach. Results The MEPI program triggered the development of new postgraduate programs thereby increasing student enrollment. This was achieved by leveraging of existing and new partnerships with other universities and differentiating the old Master in Public Health into specialized curriculum. Furthermore, the MEPI program improved the capacity and quality of training by facilitating installation and integration of new technology such as the eGranary digital library, E-learning methods and clinical skills laboratory into the Schools. This technology enabled easy access to relevant data or information, quicker turn around of experiments and enhanced data recording, display and analysis features for experiments. The program also facilitated transforming of the academic environment into a more conducive work place through strengthening the Staff Development program and support towards research activities. These activities stimulated work motivation and interest in research by faculty. Meanwhile, these processes were inhibited by the inability to upload all courses on to Moodle as well as inadequate operating procedures and feedback mechanisms for the Moodle. Conclusion Expansion and improvement in training processes for health care workers requires targeted investment within medical institutions and strengthening local and international partnerships. PMID:28819513

Strengthening and expanding the capacity of health worker education in Zambia.

PubMed

Michelo, Charles; Zulu, Joseph Mumba; Simuyemba, Moses; Andrews, Benjamin; Katubulushi, Max; Chi, Benjamin; Njelesani, Evariste; Vwalika, Bellington; Bowa, Kasonde; Maimbolwa, Margaret; Chipeta, James; Goma, Fastone; Nzala, Selestine; Banda, Sekelani; Mudenda, John; Ahmed, Yusuf; Hachambwa, Lotti; Wilson, Craig; Vermund, Sten; Mulla, Yakub

2017-01-01

Zambia is facing a chronic shortage of health care workers. The paper aimed at understanding how the Medical Education Partnership Initiative (MEPI) program facilitated strengthening and expanding of the national capacity and quality of medical education as well as processes for retaining faculty in Zambia. Data generated through documentary review, key informant interviews and observations were analyzed using a thematic approach. The MEPI program triggered the development of new postgraduate programs thereby increasing student enrollment. This was achieved by leveraging of existing and new partnerships with other universities and differentiating the old Master in Public Health into specialized curriculum. Furthermore, the MEPI program improved the capacity and quality of training by facilitating installation and integration of new technology such as the eGranary digital library, E-learning methods and clinical skills laboratory into the Schools. This technology enabled easy access to relevant data or information, quicker turn around of experiments and enhanced data recording, display and analysis features for experiments. The program also facilitated transforming of the academic environment into a more conducive work place through strengthening the Staff Development program and support towards research activities. These activities stimulated work motivation and interest in research by faculty. Meanwhile, these processes were inhibited by the inability to upload all courses on to Moodle as well as inadequate operating procedures and feedback mechanisms for the Moodle. Expansion and improvement in training processes for health care workers requires targeted investment within medical institutions and strengthening local and international partnerships.

Deterministic binary vectors for efficient automated indexing of MEDLINE/PubMed abstracts.

PubMed

Wahle, Manuel; Widdows, Dominic; Herskovic, Jorge R; Bernstam, Elmer V; Cohen, Trevor

2012-01-01

The need to maintain accessibility of the biomedical literature has led to development of methods to assist human indexers by recommending index terms for newly encountered articles. Given the rapid expansion of this literature, it is essential that these methods be scalable. Document vector representations are commonly used for automated indexing, and Random Indexing (RI) provides the means to generate them efficiently. However, RI is difficult to implement in real-world indexing systems, as (1) efficient nearest-neighbor search requires retaining all document vectors in RAM, and (2) it is necessary to maintain a store of randomly generated term vectors to index future documents. Motivated by these concerns, this paper documents the development and evaluation of a deterministic binary variant of RI. The increased capacity demonstrated by binary vectors has implications for information retrieval, and the elimination of the need to retain term vectors facilitates distributed implementations, enhancing the scalability of RI.

Deterministic Binary Vectors for Efficient Automated Indexing of MEDLINE/PubMed Abstracts

PubMed Central

Wahle, Manuel; Widdows, Dominic; Herskovic, Jorge R.; Bernstam, Elmer V.; Cohen, Trevor

2012-01-01

The need to maintain accessibility of the biomedical literature has led to development of methods to assist human indexers by recommending index terms for newly encountered articles. Given the rapid expansion of this literature, it is essential that these methods be scalable. Document vector representations are commonly used for automated indexing, and Random Indexing (RI) provides the means to generate them efficiently. However, RI is difficult to implement in real-world indexing systems, as (1) efficient nearest-neighbor search requires retaining all document vectors in RAM, and (2) it is necessary to maintain a store of randomly generated term vectors to index future documents. Motivated by these concerns, this paper documents the development and evaluation of a deterministic binary variant of RI. The increased capacity demonstrated by binary vectors has implications for information retrieval, and the elimination of the need to retain term vectors facilitates distributed implementations, enhancing the scalability of RI. PMID:23304369

pYEMF, a pUC18-derived XcmI T-vector for efficient cloning of PCR products.

PubMed

Gu, Jingsong; Ye, Chunjiang

2011-03-01

A 1330-bp DNA sequence with two XcmI cassettes was inserted into pUC18 to construct an efficient XcmI T-vector parent plasmid, pYEMF. The large size of the inserted DNA fragment improved T-vector cleavage efficiency, and guaranteed good separation of the molecular components after restriction digestion. The pYEMF-T-vector generated from parent plasmid pYEMF permits blue/white colony screening; cloning efficiency analysis showed that most white colonies (>75%) were putative transformants which carried the cloning product. The sequence analysis and design approach presented here will facilitate applications in the fields of molecular biology and genetic engineering.

Dorsolateral Striatal Lesions Impair Navigation Based on Landmark-Goal Vectors but Facilitate Spatial Learning Based on a "Cognitive Map"

ERIC Educational Resources Information Center

Kosaki, Yutaka; Poulter, Steven L.; Austen, Joe M.; McGregor, Anthony

2015-01-01

In three experiments, the nature of the interaction between multiple memory systems in rats solving a variation of a spatial task in the water maze was investigated. Throughout training rats were able to find a submerged platform at a fixed distance and direction from an intramaze landmark by learning a landmark-goal vector. Extramaze cues were…

Barriers and facilitators to development of standard treatment guidelines in India.

PubMed

Sharma, Sangeeta; Sethi, Gulshan R; Gupta, Usha; Chaudhury, Ranjit Roy

2015-01-01

This paper describes 15 years' experience of the development process of the first set of comprehensive standard treatment guidelines (STGs) for India and their adoption or adaptation by various state governments. The aim is to shorten the learning curve for those embarking on a similar exercise, given the key role of high-quality STGs that are accepted by the clinical community in furthering universal health coverage. The main overall obstacles to STG development are: (i) weak understanding of the concept; (ii) lack of time, enthusiasm and availability of local expertise; and (iii) managing consensus between specialists and generalists. Major concerns to prescribers are: encroachment on professional autonomy, loss of treating the patient as an individual and applying the same standards at all levels of health care. Processes to address these challenges are described. At the policy level, major threats to successful completion and focused implementation are: frequent changes in governance, shifts in priorities and discontinuity. In the authors' experience, compared with each state developing their own STGs afresh, adaptation of pre-existing valid guidelines after an active adaptation process involving local clinical leaders is not only simpler and quicker but also establishes local ownership and facilitates acceptance of a quality document. Executive orders and in-service sensitization programmes to introduce STGs further enhance their adoption in clinical practice.

Development and evaluation of an instrumented linkage system for total knee surgery.

PubMed

Walker, Peter S; Wei, Chih-Shing; Forman, Rachel E; Balicki, M A

2007-10-01

The principles and application of total knee surgery using optical tracking have been well demonstrated, but electromagnetic tracking may offer further advantages. We asked whether an instrumented linkage that attaches directly to the bone can maintain the accuracy of the optical and electromagnetic systems but be quicker, more convenient, and less expensive to use. Initial testing using a table-mounted digitizer to navigate a drill guide for placing pins to mount a cutting guide demonstrated the feasibility in terms of access and availability. A first version (called the Mark 1) instrumented linkage designed to fix directly to the bone was constructed and software was written to carry out a complete total knee replacement procedure. The results showed the system largely fulfilled these goals, but some surgeons found that using a visual display for pin placement was difficult and time consuming. As a result, a second version of a linkage system (called the K-Link) was designed to further develop the concept. User-friendly flexible software was developed for facilitating each step quickly and accurately while the placement of cutting guides was facilitated. We concluded that an instrumented linkage system could be a useful and potentially lower-cost option to the current systems for total knee replacement and could possibly have application to other surgical procedures.

The Anopheles gambiae transcriptome - a turning point for malaria control.

PubMed

Domingos, A; Pinheiro-Silva, R; Couto, J; do Rosário, V; de la Fuente, J

2017-04-01

Mosquitoes are important vectors of several pathogens and thereby contribute to the spread of diseases, with social, economic and public health impacts. Amongst the approximately 450 species of Anopheles, about 60 are recognized as vectors of human malaria, the most important parasitic disease. In Africa, Anopheles gambiae is the main malaria vector mosquito. Current malaria control strategies are largely focused on drugs and vector control measures such as insecticides and bed-nets. Improvement of current, and the development of new, mosquito-targeted malaria control methods rely on a better understanding of mosquito vector biology. An organism's transcriptome is a reflection of its physiological state and transcriptomic analyses of different conditions that are relevant to mosquito vector competence can therefore yield important information. Transcriptomic analyses have contributed significant information on processes such as blood-feeding parasite-vector interaction, insecticide resistance, and tissue- and stage-specific gene regulation, thereby facilitating the path towards the development of new malaria control methods. Here, we discuss the main applications of transcriptomic analyses in An. gambiae that have led to a better understanding of mosquito vector competence. © 2017 The Royal Entomological Society.

Re-engineering adenovirus vector systems to enable high-throughput analyses of gene function.

PubMed

Stanton, Richard J; McSharry, Brian P; Armstrong, Melanie; Tomasec, Peter; Wilkinson, Gavin W G

2008-12-01

With the enhanced capacity of bioinformatics to interrogate extensive banks of sequence data, more efficient technologies are needed to test gene function predictions. Replication-deficient recombinant adenovirus (Ad) vectors are widely used in expression analysis since they provide for extremely efficient expression of transgenes in a wide range of cell types. To facilitate rapid, high-throughput generation of recombinant viruses, we have re-engineered an adenovirus vector (designated AdZ) to allow single-step, directional gene insertion using recombineering technology. Recombineering allows for direct insertion into the Ad vector of PCR products, synthesized sequences, or oligonucleotides encoding shRNAs without requirement for a transfer vector Vectors were optimized for high-throughput applications by making them "self-excising" through incorporating the I-SceI homing endonuclease into the vector removing the need to linearize vectors prior to transfection into packaging cells. AdZ vectors allow genes to be expressed in their native form or with strep, V5, or GFP tags. Insertion of tetracycline operators downstream of the human cytomegalovirus major immediate early (HCMV MIE) promoter permits silencing of transgenes in helper cells expressing the tet repressor thus making the vector compatible with the cloning of toxic gene products. The AdZ vector system is robust, straightforward, and suited to both sporadic and high-throughput applications.

High-Throughput Functional Validation of Progression Drivers in Lung Adenocarcinoma

DTIC Science & Technology

2013-09-01

2) a novel molecular barcoding approach that facilitates cost- effective detection of driver events following in vitro and in vivo functional screens...aberration construction pipeline, which we named High-Throughput 3 Mutagenesis and Molecular Barcoding (HiTMMoB; Fig.1). We have therefore been able...lentiviral vector specially constructed for this project. This vector is compatible with our flexible molecular barcoding technology (Fig. 1), thus each

Dorsolateral striatal lesions impair navigation based on landmark-goal vectors but facilitate spatial learning based on a “cognitive map”

PubMed Central

Poulter, Steven L.; Austen, Joe M.

2015-01-01

In three experiments, the nature of the interaction between multiple memory systems in rats solving a variation of a spatial task in the water maze was investigated. Throughout training rats were able to find a submerged platform at a fixed distance and direction from an intramaze landmark by learning a landmark-goal vector. Extramaze cues were also available for standard place learning, or “cognitive mapping,” but these cues were valid only within each session, as the position of the platform moved around the pool between sessions together with the intramaze landmark. Animals could therefore learn the position of the platform by taking the consistent vector from the landmark across sessions or by rapidly encoding the new platform position on each session with reference to the extramaze cues. Excitotoxic lesions of the dorsolateral striatum impaired vector-based learning but facilitated cognitive map-based rapid place learning when the extramaze cues were relatively poor (Experiment 1) but not when they were more salient (Experiments 2 and 3). The way the lesion effects interacted with cue availability is consistent with the idea that the memory systems involved in the current navigation task are functionally cooperative yet associatively competitive in nature. PMID:25691518

Declining Prevalence of Disease Vectors Under Climate Change

NASA Astrophysics Data System (ADS)

Escobar, Luis E.; Romero-Alvarez, Daniel; Leon, Renato; Lepe-Lopez, Manuel A.; Craft, Meggan E.; Borbor-Cordova, Mercy J.; Svenning, Jens-Christian

2016-12-01

More than half of the world population is at risk of vector-borne diseases including dengue fever, chikungunya, zika, yellow fever, leishmaniasis, chagas disease, and malaria, with highest incidences in tropical regions. In Ecuador, vector-borne diseases are present from coastal and Amazonian regions to the Andes Mountains; however, a detailed characterization of the distribution of their vectors has never been carried out. We estimate the distribution of 14 vectors of the above vector-borne diseases under present-day and future climates. Our results consistently suggest that climate warming is likely threatening some vector species with extinction, locally or completely. These results suggest that climate change could reduce the burden of specific vector species. Other vector species are likely to shift and constrain their geographic range to the highlands in Ecuador potentially affecting novel areas and populations. These forecasts show the need for development of early prevention strategies for vector species currently absent in areas projected as suitable under future climate conditions. Informed interventions could reduce the risk of human exposure to vector species with distributional shifts, in response to current and future climate changes. Based on the mixed effects of future climate on human exposure to disease vectors, we argue that research on vector-borne diseases should be cross-scale and include climatic, demographic, and landscape factors, as well as forces facilitating disease transmission at fine scales.

Helminths as vectors of pathogens in vertebrate hosts: a theoretical approach.

PubMed

Perkins, Sarah E; Fenton, Andy

2006-07-01

Pathogens frequently use vectors to facilitate transmission between hosts and, for vertebrate hosts, the vectors are typically ectoparasitic arthropods. However, other parasites that are intimately associated with their hosts may also be ideal candidate vectors; namely the parasitic helminths. Here, we present empirical evidence that helminth vectoring of pathogens occurs in a range of vertebrate systems by a variety of helminth taxa. Using a novel theoretical framework we explore the dynamics of helminth vectoring and determine which host-helminth-pathogen characteristics may favour the evolution of helminth vectoring. We use two theoretical models: the first is a population dynamic model amalgamated from standard macro- and microparasite models, which serves as a framework for investigation of within-host interactions between co-infecting pathogens and helminths. The second is an evolutionary model, which we use to predict the ecological conditions under which we would expect helminth vectoring to evolve. We show that, like arthropod vectors, helminth vectors increase pathogen fitness. However, unlike arthropod vectors, helminth vectoring increases the pathogenic impact on the host and may allow the evolution of high pathogen virulence. We show that concomitant infection of a host with a helminth and pathogen are not necessarily independent of one another, due to helminth vectoring of microparasites, with profound consequences for pathogen persistence and the impact of disease on the host population.

Generation of Marker- and/or Backbone-Free Transgenic Wheat Plants via Agrobacterium-Mediated Transformation.

PubMed

Wang, Gen-Ping; Yu, Xiu-Dao; Sun, Yong-Wei; Jones, Huw D; Xia, Lan-Qin

2016-01-01

Horizontal transfer of antibiotic resistance genes to animals and vertical transfer of herbicide resistance genes to the weedy relatives are perceived as major biosafety concerns in genetically modified (GM) crops. In this study, five novel vectors which used gusA and bar as a reporter gene and a selection marker gene, respectively, were constructed based on the pCLEAN dual binary vector system. Among these vectors, 1G7B and 5G7B carried two T-DNAs located on two respective plasmids with 5G7B possessing an additional virGwt gene. 5LBTG154 and 5TGTB154 carried two T-DNAs in the target plasmid with either one or double right borders, and 5BTG154 carried the selectable marker gene on the backbone outside of the T-DNA left border in the target plasmid. In addition, 5BTG154, 5LBTG154, and 5TGTB154 used pAL154 as a helper plasmid which contains Komari fragment to facilitate transformation. These five dual binary vector combinations were transformed into Agrobacterium strain AGL1 and used to transform durum wheat cv Stewart 63. Evaluation of the co-transformation efficiencies, the frequencies of marker-free transgenic plants, and integration of backbone sequences in the obtained transgenic lines indicated that two vectors (5G7B and 5TGTB154) were more efficient in generating marker-free transgenic wheat plants with no or minimal integration of backbone sequences in the wheat genome. The vector series developed in this study for generation of marker- and/or backbone-free transgenic wheat plants via Agrobacterium -mediated transformation will be useful to facilitate the creation of "clean" GM wheat containing only the foreign genes of agronomic importance.

Towards an integrated approach in surveillance of vector-borne diseases in Europe

PubMed Central

2011-01-01

Vector borne disease (VBD) emergence is a complex and dynamic process. Interactions between multiple disciplines and responsible health and environmental authorities are often needed for an effective early warning, surveillance and control of vectors and the diseases they transmit. To fully appreciate this complexity, integrated knowledge about the human and the vector population is desirable. In the current paper, important parameters and terms of both public health and medical entomology are defined in order to establish a common language that facilitates collaboration between the two disciplines. Special focus is put on the different VBD contexts with respect to the current presence or absence of the disease, the pathogen and the vector in a given location. Depending on the context, whether a VBD is endemic or not, surveillance activities are required to assess disease burden or threat, respectively. Following a decision for action, surveillance activities continue to assess trends. PMID:21967706

Viral Vectors for In Vivo Gene Transfer in Parkinson’s disease: Properties and Clinical Grade Production

PubMed Central

Burger, Corinna; Snyder, Richard O.

2009-01-01

Because Parkinson’s disease is a progressive degenerative disorder that is mainly confined to the basal ganglia, gene transfer to deliver therapeutic molecules is an attractive treatment avenue. The present review focuses on direct in vivo gene transfer vectors that have been developed to a degree that they have been successfully used in animal model of Parkinson’s disease. Accordingly, the properties of recombinant adenovirus, recombinant adeno-associated virus, herpes simplex virus, and lentivirus are described and contrasted. In order for viral vectors to be developed into clinical grade reagents, they must be manufactured and tested to precise regulatory standards. Indeed, clinical lots of viral vectors can be produced in compliance with current Good Manufacturing Practices (cGMPs) regulations using industry accepted manufacturing methodologies, manufacturing controls, and quality systems. The viral vector properties themselves combined with physiological product formulations facilitate long-term storage and direct in vivo administration. PMID:17916354

The emerging contribution of social wasps to grape rot disease ecology

PubMed Central

Boyden, Sean D.; Soriano, Jonathan-Andrew N.; Corey, Tyler B.; Leff, Jonathan W.; Fierer, Noah; Starks, Philip T.

2017-01-01

Grape sour (bunch) rot is a polymicrobial disease of vineyards that causes millions of dollars in lost revenue per year due to decreased quality of grapes and resultant wine. The disease is associated with damaged berries infected with a community of acetic acid bacteria, yeasts, and filamentous fungi that results in rotting berries with high amounts of undesirable volatile acidity. Many insect species cause the initial grape berry damage that can lead to this disease, but most studies have focused on the role of fruit flies in facilitating symptoms and vectoring the microorganisms of this disease complex. Like fruit flies, social wasps are abundant in vineyards where they feed on ripe berries and cause significant damage, while also dispersing yeasts involved in wine fermentation. Despite this, their possible role in disease facilitation and dispersal of grape rots has not been explored. We tested the hypothesis that the paper wasp Polistes dominulus could facilitate grape sour rot in the absence of other insect vectors. Using marker gene sequencing we characterized the bacterial and fungal community of wild-caught adults. We used a sterilized foraging arena to determine if these wasps transfer viable microorganisms when foraging. We then tested if wasps harboring their native microbial community, or those inoculated with sour rot, had an effect on grape sour rot incidence and severity using a laboratory foraging arena. We found that all wasps harbor some portion of the sour rot microbial community and that they have the ability to transfer viable microorganisms when foraging. Foraging by inoculated and uninoculated wasps led to an increase in berry rot disease symptom severity and incidence. Our results indicate that paper wasps can facilitate sour rot diseases in the absence of other vectors and that the mechanism of this facilitation may include both increasing host susceptibility and transmitting these microbial communities to the grapes. Social wasps are understudied but relevant players in the sour rot ecology of vineyards. PMID:28462032

The AIST Managed Cloud Environment

NASA Astrophysics Data System (ADS)

Cook, S.

2016-12-01

ESTO is currently in the process of developing and implementing the AIST Managed Cloud Environment (AMCE) to offer cloud computing services to ESTO-funded PIs to conduct their project research. AIST will provide projects access to a cloud computing framework that incorporates NASA security, technical, and financial standards, on which project can freely store, run, and process data. Currently, many projects led by research groups outside of NASA do not have the awareness of requirements or the resources to implement NASA standards into their research, which limits the likelihood of infusing the work into NASA applications. Offering this environment to PIs will allow them to conduct their project research using the many benefits of cloud computing. In addition to the well-known cost and time savings that it allows, it also provides scalability and flexibility. The AMCE will facilitate infusion and end user access by ensuring standardization and security. This approach will ultimately benefit ESTO, the science community, and the research, allowing the technology developments to have quicker and broader applications.

Kinesthetic Imagery Provides Additive Benefits to Internal Visual Imagery on Slalom Task Performance.

PubMed

Callow, Nichola; Jiang, Dan; Roberts, Ross; Edwards, Martin G

2017-02-01

Recent brain imaging research demonstrates that the use of internal visual imagery (IVI) or kinesthetic imagery (KIN) activates common and distinct brain areas. In this paper, we argue that combining the imagery modalities (IVI and KIN) will lead to a greater cognitive representation (with more brain areas activated), and this will cause a greater slalom-based motor performance compared with using IVI alone. To examine this assertion, we randomly allocated 56 participants to one of the three groups: IVI, IVI and KIN, or a math control group. Participants performed a slalom-based driving task in a driving simulator, with average lap time used as a measure of performance. Results revealed that the IVI and KIN group achieved significantly quicker lap times than the IVI and the control groups. The discussion includes a theoretical advancement on why the combination of imagery modalities might facilitate performance, with links made to the cognitive neuroscience literature and applied practice.

The AMCE (AIST Managed Cloud Environment)

NASA Astrophysics Data System (ADS)

Cook, S.

2017-12-01

ESTO has developed and implemented the AIST Managed Cloud Environment (AMCE) to offer cloud computing services to SMD-funded PIs to conduct their project research. AIST will provide projects access to a cloud computing framework that incorporates NASA security, technical, and financial standards, on which project can freely store, run, and process data. Currently, many projects led by research groups outside of NASA do not have the awareness of requirements or the resources to implement NASA standards into their research, which limits the likelihood of infusing the work into NASA applications. Offering this environment to PIs allows them to conduct their project research using the many benefits of cloud computing. In addition to the well-known cost and time savings that it allows, it also provides scalability and flexibility. The AMCE facilitates infusion and end user access by ensuring standardization and security. This approach will ultimately benefit ESTO, the science community, and the research, allowing the technology developments to have quicker and broader applications.

A case report of laparoscopic duodenal atresia repair in a neonate using a novel miniature stapling device.

PubMed

Boo, Yoon Jung; Gödeke, Jan; Engel, Veronika; Muensterer, Oliver J

2017-01-01

Laparoscopic duodenal atresia repair is an advanced procedure performed in select pediatric surgical centers. Recently, sub-6mm endosurgical staplers have been introduced, facilitating and accelerating the creation of intracorporeal intestinal anastomoses. We performed a laparoscopic duodenojejunostomy in a one-day-old child with duodenal atresia due to annular pancreas using a novel 5.8mm articulating endostapler with excellent outcome. The technical details are reported. Laparoscopic duodenojejunostomy is a technically demanding procedure due to difficulty in hand-sewn anastomosis in a small and restricted space. With this novel 5.8mm articulating endostapler, we were able to perform a quicker and easier anastomosis. We report a case of laparoscopic duodenal atresia repair in a neonate using a novel miniature stapling device. This new technique is a safe, quick and easier way to perform laparoscopic duodenal atresia repair. Copyright © 2016 The Author(s). Published by Elsevier Ltd.. All rights reserved.

Integration process of fermentation and liquid biphasic flotation for lipase separation from Burkholderia cepacia.

PubMed

Sankaran, Revathy; Show, Pau Loke; Lee, Sze Ying; Yap, Yee Jiun; Ling, Tau Chuan

2018-02-01

Liquid Biphasic Flotation (LBF) is an advanced recovery method that has been effectively applied for biomolecules extraction. The objective of this investigation is to incorporate the fermentation and extraction process of lipase from Burkholderia cepacia using flotation system. Initial study was conducted to compare the performance of bacteria growth and lipase production using flotation and shaker system. From the results obtained, bacteria shows quicker growth and high lipase yield via flotation system. Integration process for lipase separation was investigated and the result showed high efficiency reaching 92.29% and yield of 95.73%. Upscaling of the flotation system exhibited consistent result with the lab-scale which are 89.53% efficiency and 93.82% yield. The combination of upstream and downstream processes in a single system enables the acceleration of product formation, improves the product yield and facilitates downstream processing. This integration system demonstrated its potential for biomolecules fermentation and separation that possibly open new opportunities for industrial production. Copyright © 2017 Elsevier Ltd. All rights reserved.

Collaborative mining and interpretation of large-scale data for biomedical research insights.

PubMed

Tsiliki, Georgia; Karacapilidis, Nikos; Christodoulou, Spyros; Tzagarakis, Manolis

2014-01-01

Biomedical research becomes increasingly interdisciplinary and collaborative in nature. Researchers need to efficiently and effectively collaborate and make decisions by meaningfully assembling, mining and analyzing available large-scale volumes of complex multi-faceted data residing in different sources. In line with related research directives revealing that, in spite of the recent advances in data mining and computational analysis, humans can easily detect patterns which computer algorithms may have difficulty in finding, this paper reports on the practical use of an innovative web-based collaboration support platform in a biomedical research context. Arguing that dealing with data-intensive and cognitively complex settings is not a technical problem alone, the proposed platform adopts a hybrid approach that builds on the synergy between machine and human intelligence to facilitate the underlying sense-making and decision making processes. User experience shows that the platform enables more informed and quicker decisions, by displaying the aggregated information according to their needs, while also exploiting the associated human intelligence.

Collaborative Mining and Interpretation of Large-Scale Data for Biomedical Research Insights

PubMed Central

Tsiliki, Georgia; Karacapilidis, Nikos; Christodoulou, Spyros; Tzagarakis, Manolis

2014-01-01

Biomedical research becomes increasingly interdisciplinary and collaborative in nature. Researchers need to efficiently and effectively collaborate and make decisions by meaningfully assembling, mining and analyzing available large-scale volumes of complex multi-faceted data residing in different sources. In line with related research directives revealing that, in spite of the recent advances in data mining and computational analysis, humans can easily detect patterns which computer algorithms may have difficulty in finding, this paper reports on the practical use of an innovative web-based collaboration support platform in a biomedical research context. Arguing that dealing with data-intensive and cognitively complex settings is not a technical problem alone, the proposed platform adopts a hybrid approach that builds on the synergy between machine and human intelligence to facilitate the underlying sense-making and decision making processes. User experience shows that the platform enables more informed and quicker decisions, by displaying the aggregated information according to their needs, while also exploiting the associated human intelligence. PMID:25268270

Oral Midazolam Sedation For Uncooperative Children In Outpatient Paedodontics: Time For Reappraisal.

PubMed

Kapur, Aditi; Jain, Kajal; Goyal, A; Mahoney, Greg

2016-01-01

Sedation is frequently desired to facilitate dental procedures in uncooperative paediatric patients. Oromucosal Midazolam sedation is a popular choice among paediatric dentists world wide due to its many advantages such as ease of administration, good efficacy, presence of reversal agents and a wide margin of safety. On the other hand, many investigators have reported that midazolam sedation may not be successful for carrying out all types of dental procedures. This may be attributed to diverse nature of various treatment plans coupled with the extent of behavioural changes in the child and operator's experience. Due to the heterogeneity involved in treatment of paediatric dental procedures, the specific indications for oral midazolam use that ensure its success rate, probably need to be defined. This may enable the clinicians to have a convenient and quicker option for managing the cases rather than facing sedation failure or at times, ending up giving general anaesthetics. This article therefore brings forth the possible causes of midazolam sedation failure and proposes a 'case selection criterion'.

Contribution of sleep to the repair of neuronal DNA double-strand breaks: evidence from flies and mice.

PubMed

Bellesi, Michele; Bushey, Daniel; Chini, Mattia; Tononi, Giulio; Cirelli, Chiara

2016-11-10

Exploration of a novel environment leads to neuronal DNA double-strand breaks (DSBs). These DSBs are generated by type 2 topoisomerase to relieve topological constrains that limit transcription of plasticity-related immediate early genes. If not promptly repaired, however, DSBs may lead to cell death. Since the induction of plasticity-related genes is higher in wake than in sleep, we asked whether it is specifically wake associated with synaptic plasticity that leads to DSBs, and whether sleep provides any selective advantage over wake in their repair. In flies and mice, we find that enriched wake, more than simply time spent awake, induces DSBs, and their repair in mice is delayed or prevented by subsequent wake. In both species the repair of irradiation-induced neuronal DSBs is also quicker during sleep, and mouse genes mediating the response to DNA damage are upregulated in sleep. Thus, sleep facilitates the repair of neuronal DSBs.

Ontology for Vector Surveillance and Management

PubMed Central

LOZANO-FUENTES, SAUL; BANDYOPADHYAY, ARITRA; COWELL, LINDSAY G.; GOLDFAIN, ALBERT; EISEN, LARS

2013-01-01

Ontologies, which are made up by standardized and defined controlled vocabulary terms and their interrelationships, are comprehensive and readily searchable repositories for knowledge in a given domain. The Open Biomedical Ontologies (OBO) Foundry was initiated in 2001 with the aims of becoming an “umbrella” for life-science ontologies and promoting the use of ontology development best practices. A software application (OBO-Edit; *.obo file format) was developed to facilitate ontology development and editing. The OBO Foundry now comprises over 100 ontologies and candidate ontologies, including the NCBI organismal classification ontology (NCBITaxon), the Mosquito Insecticide Resistance Ontology (MIRO), the Infectious Disease Ontology (IDO), the IDOMAL malaria ontology, and ontologies for mosquito gross anatomy and tick gross anatomy. We previously developed a disease data management system for dengue and malaria control programs, which incorporated a set of information trees built upon ontological principles, including a “term tree” to promote the use of standardized terms. In the course of doing so, we realized that there were substantial gaps in existing ontologies with regards to concepts, processes, and, especially, physical entities (e.g., vector species, pathogen species, and vector surveillance and management equipment) in the domain of surveillance and management of vectors and vector-borne pathogens. We therefore produced an ontology for vector surveillance and management, focusing on arthropod vectors and vector-borne pathogens with relevance to humans or domestic animals, and with special emphasis on content to support operational activities through inclusion in databases, data management systems, or decision support systems. The Vector Surveillance and Management Ontology (VSMO) includes >2,200 unique terms, of which the vast majority (>80%) were newly generated during the development of this ontology. One core feature of the VSMO is the linkage, through the has_vector relation, of arthropod species to the pathogenic microorganisms for which they serve as biological vectors. We also recognized and addressed a potential roadblock for use of the VSMO by the vector-borne disease community: the difficulty in extracting information from OBO-Edit ontology files (*.obo files) and exporting the information to other file formats. A novel ontology explorer tool was developed to facilitate extraction and export of information from the VSMO *.obo file into lists of terms and their associated unique IDs in *.txt or *.csv file formats. These lists can then be imported into a database or data management system for use as select lists with predefined terms. This is an important step to ensure that the knowledge contained in our ontology can be put into practical use. PMID:23427646

Ontology for vector surveillance and management.

PubMed

Lozano-Fuentes, Saul; Bandyopadhyay, Aritra; Cowell, Lindsay G; Goldfain, Albert; Eisen, Lars

2013-01-01

Ontologies, which are made up by standardized and defined controlled vocabulary terms and their interrelationships, are comprehensive and readily searchable repositories for knowledge in a given domain. The Open Biomedical Ontologies (OBO) Foundry was initiated in 2001 with the aims of becoming an "umbrella" for life-science ontologies and promoting the use of ontology development best practices. A software application (OBO-Edit; *.obo file format) was developed to facilitate ontology development and editing. The OBO Foundry now comprises over 100 ontologies and candidate ontologies, including the NCBI organismal classification ontology (NCBITaxon), the Mosquito Insecticide Resistance Ontology (MIRO), the Infectious Disease Ontology (IDO), the IDOMAL malaria ontology, and ontologies for mosquito gross anatomy and tick gross anatomy. We previously developed a disease data management system for dengue and malaria control programs, which incorporated a set of information trees built upon ontological principles, including a "term tree" to promote the use of standardized terms. In the course of doing so, we realized that there were substantial gaps in existing ontologies with regards to concepts, processes, and, especially, physical entities (e.g., vector species, pathogen species, and vector surveillance and management equipment) in the domain of surveillance and management of vectors and vector-borne pathogens. We therefore produced an ontology for vector surveillance and management, focusing on arthropod vectors and vector-borne pathogens with relevance to humans or domestic animals, and with special emphasis on content to support operational activities through inclusion in databases, data management systems, or decision support systems. The Vector Surveillance and Management Ontology (VSMO) includes >2,200 unique terms, of which the vast majority (>80%) were newly generated during the development of this ontology. One core feature of the VSMO is the linkage, through the has vector relation, of arthropod species to the pathogenic microorganisms for which they serve as biological vectors. We also recognized and addressed a potential roadblock for use of the VSMO by the vector-borne disease community: the difficulty in extracting information from OBO-Edit ontology files (*.obo files) and exporting the information to other file formats. A novel ontology explorer tool was developed to facilitate extraction and export of information from the VSMO*.obo file into lists of terms and their associated unique IDs in *.txt or *.csv file formats. These lists can then be imported into a database or data management system for use as select lists with predefined terms. This is an important step to ensure that the knowledge contained in our ontology can be put into practical use.

A vectorization of the Hess McDonnell Douglas potential flow program NUED for the STAR-100 computer

NASA Technical Reports Server (NTRS)

Boney, L. R.; Smith, R. E., Jr.

1979-01-01

The computer program NUED for analyzing potential flow about arbitrary three dimensional lifting bodies using the panel method was modified to use vector operations and run on the STAR-100 computer. A high speed of computation and ability to approximate the body surface with a large number of panels are characteristics of NUEDV. The new program shows that vector operations can be readily implemented in programs of this type to increase the computational speed on the STAR-100 computer. The virtual memory architecture of the STAR-100 facilitates the use of large numbers of panels to approximate the body surface.

Return of epidemic dengue in the United States: implications for the public health practitioner.

PubMed

Bouri, Nidhi; Sell, Tara Kirk; Franco, Crystal; Adalja, Amesh A; Henderson, D A; Hynes, Noreen A

2012-01-01

Conditions that facilitate sustained dengue transmission exist in the United States, and outbreaks have occurred during the past decade in Texas, Hawaii, and Florida. More outbreaks can also be expected in years to come. To combat dengue, medical and public health practitioners in areas with mosquito vectors that are competent to transmit the virus must be aware of the threat of reemergent dengue, and the need for early reporting and control to reduce the impact of dengue outbreaks. Comprehensive dengue control includes human and vector surveillance, vector management programs, and community engagement efforts. Public health, medical, and vector-control communities must collaborate to prevent and control disease spread. Policy makers should understand the role of mosquito abatement and community engagement in the prevention and control of the disease.

Gene silencing in Escherichia coli using antisense RNAs expressed from doxycycline-inducible vectors.

PubMed

Nakashima, N; Tamura, T

2013-06-01

Here, we report on the construction of doxycycline (tetracycline analogue)-inducible vectors that express antisense RNAs in Escherichia coli. Using these vectors, the expression of genes of interest can be silenced conditionally. The expression of antisense RNAs from the vectors was more tightly regulated than the previously constructed isopropyl-β-D-galactopyranoside-inducible vectors. Furthermore, expression levels of antisense RNAs were enhanced by combining the doxycycline-inducible promoter with the T7 promoter-T7 RNA polymerase system; the T7 RNA polymerase gene, under control of the doxycycline-inducible promoter, was integrated into the lacZ locus of the genome without leaving any antibiotic marker. These vectors are useful for investigating gene functions or altering cell phenotypes for biotechnological and industrial applications. A gene silencing method using antisense RNAs in Escherichia coli is described, which facilitates the investigation of bacterial gene function. In particular, the method is suitable for comprehensive analyses or phenotypic analyses of genes essential for growth. Here, we describe expansion of vector variations for expressing antisense RNAs, allowing choice of a vector appropriate for the target genes or experimental purpose. © 2013 The Society for Applied Microbiology.

[What makes an insect a vector?].

PubMed

Kampen, Helge

2009-01-01

Blood-feeding insects transmit numerous viruses, bacteria, protozoans and helminths to vertebrates. The developmental cycles of the microorganisms in their vectors and the mechanisms of transmission are generally extremely complex and the result of a long-lasting coevolution of vector and vectored pathogen based on mutual adaptation. The conditions necessary for an insect to become a vector are multiple but require an innate vector competence as a genetic basis. Next to the vector competence plenty of entomological, ecological and pathogen-related factors are decisive, given the availability of infection sources. The various modes of pathogen transmission by vectors are connected to the developmental routes of the microorganisms in their vectors. In particular, pathogens transmitted by saliva encounter a lot of cellular and acellular barriers during their migration from the insect's midgut through the hemocele into the salivary fluid, including components of the insect's immune system. With regard to intracellular development, receptor-mediated invasion mechanisms are of relevance. As an environmental factor, the temperature has a paramount impact on the vectorial roles of hematophagous insects. Not only has it a considerable influence on the duration of a pathogen's development in its vector (extrinsic incubation period) but it can render putatively vector-incompetent insects to vectors ("leaky gut" phenomenon). Equally crucial are behavioural aspects of both the insect and the pathogen such as blood host preferences, seasonal appearance and circadian biting activity on the vector's side and diurnal/nocturnal periodicity on the pathogen's side which facilitate a contact in the first place.

A web implementation: the good and the not-so-good.

PubMed

Bergsneider, C; Piraino, D; Fuerst, M

2001-06-01

E-commerce, e-mail, e-greeting, e-this, and e-that everywhere you turn there is a new "e" word for an internet or Web application. We, at the Cleveland Clinic Foundation, have been "e-nlightened" and will discuss in this report the implementation of a web-based radiology information system (RIS) in our radiology division or "e-radiology" division. The application, IDXRad Version 10.0 from IDX Corp, Burlington, VT, is in use at the Cleveland Clinic Foundation and has both intranet (for use in Radiology) and internet (referring physician viewing) modules. We will concentrate on the features of using a web browser for the application's front-end, including easy prototyping for screen review, easier mock-ups of demonstrations by vendors and developers, and easier training as more people become web-addicted. Project communication can be facilitated with an internal project web page, and use of the web browser can accommodate quicker turnaround of software upgrades as the software code is centrally located. Compared with other technologies, including client/server, there is a smaller roll out cost when using a standard web browser. However, the new technology requires a change and changes are never implemented without challenges. A seasoned technologist using a legacy system can enter data quicker using function keys than using a graphical user interface and pointing and clicking through a series of pop-up windows. Also, effective use of a web browser depends on intuitive design for it to be easily implemented and accepted by the user. Some software packages will not work on both of the popular web browsers and then are tailored to specific release levels. As computer-based patient records become a standard, patient confidentiality must be enforced. The technical design and application security features that support the web-based software package will be discussed. Also web technologies have their own implementation issues.

A new series of yeast shuttle vectors for the recovery and identification of multiple plasmids from Saccharomyces cerevisiae.

PubMed

Frazer, LilyAnn Novak; O'Keefe, Raymond T

2007-09-01

The availability of Saccharomyces cerevisiae yeast strains with multiple auxotrophic markers allows the stable introduction and selection of more than one yeast shuttle vector containing marker genes that complement the auxotrophic markers. In certain experimental situations there is a need to recover more than one shuttle vector from yeast. To facilitate the recovery and identification of multiple plasmids from S. cerevisiae, we have constructed a series of plasmids based on the pRS series of yeast shuttle vectors. Bacterial antibiotic resistance genes to chloramphenicol, kanamycin and zeocin have been combined with the yeast centromere sequence (CEN6), the autonomously replicating sequence (ARSH4) and one of the four yeast selectable marker genes (HIS3, TRP1, LEU2 or URA3) from the pRS series of vectors. The 12 plasmids produced differ in antibiotic resistance and yeast marker gene within the backbone of the multipurpose plasmid pBluescript II. The newly constructed vectors show similar mitotic stability to the original pRS vectors. In combination with the ampicillin-resistant pRS series of yeast shuttle vectors, these plasmids now allow the recovery and identification in bacteria of up to four different vectors from S. cerevisiae. Copyright (c) 2007 John Wiley & Sons, Ltd.

EMMA: An Extensible Mammalian Modular Assembly Toolkit for the Rapid Design and Production of Diverse Expression Vectors.

PubMed

Martella, Andrea; Matjusaitis, Mantas; Auxillos, Jamie; Pollard, Steven M; Cai, Yizhi

2017-07-21

Mammalian plasmid expression vectors are critical reagents underpinning many facets of research across biology, biomedical research, and the biotechnology industry. Traditional cloning methods often require laborious manual design and assembly of plasmids using tailored sequential cloning steps. This process can be protracted, complicated, expensive, and error-prone. New tools and strategies that facilitate the efficient design and production of bespoke vectors would help relieve a current bottleneck for researchers. To address this, we have developed an extensible mammalian modular assembly kit (EMMA). This enables rapid and efficient modular assembly of mammalian expression vectors in a one-tube, one-step golden-gate cloning reaction, using a standardized library of compatible genetic parts. The high modularity, flexibility, and extensibility of EMMA provide a simple method for the production of functionally diverse mammalian expression vectors. We demonstrate the value of this toolkit by constructing and validating a range of representative vectors, such as transient and stable expression vectors (transposon based vectors), targeting vectors, inducible systems, polycistronic expression cassettes, fusion proteins, and fluorescent reporters. The method also supports simple assembly combinatorial libraries and hierarchical assembly for production of larger multigenetic cargos. In summary, EMMA is compatible with automated production, and novel genetic parts can be easily incorporated, providing new opportunities for mammalian synthetic biology.

Low-volume application by mist-blower compared with conventional compression sprayer treatment of houses with residual pyrethroid to control the malaria vector Anopheles albimanus in Mexico.

PubMed

Villarreal, C; Rodriguez, M H; Bown, D N; Arredondo-Jiménez, J I

1995-04-01

Village-scale trials were carried out in southern Mexico to compare the efficacy of indoor-spraying of the pyrethroid insecticide lambda-cyhalothrin applied either as low-volume (LV) aqueous emulsion or as wettable-powder (WP) aqueous suspension for residual control of the principal coastal malaria vector Anopheles albimanus. Three indoor spray rounds were conducted at 3-month intervals using back-pack mist-blowers to apply lambda-cyhalothrin 12.5 mg a.i./m2 by LV, whereas the WP was applied by conventional compression sprayer at a mean rate of 26.5 mg a.i./m2. Both treatments caused mosquito mortality indoors and outdoors (collected inside house curtains) as a result of contact with treated surfaces before and after feeding, but had no significant impact on overall population density of An. albimanus resting indoors or assessed by human bait collections. Contact bioassays showed that WP and LV treatments with lambda-cyhalothrin were effective for 12-20 weeks (> 75% mortality) without causing excito-repellency. Compared to the WP treatment (8 houses/man/day), LV treatment (25 houses/man/day) was more than 3 times quicker per house, potentially saving 68% of labour costs. This is offset, however, by the much lower unit price of a compression sprayer (e.g. Hudson 'X-pert' at US$120) than a mist-blower (e.g. 'Super Jolly' at US$350), and higher running costs for LV applications. It was calculated, therefore, that LV becomes more economical than WP after 18.8 treatments/100 houses/10 men at equivalent rates of application, or after 7.6 spray rounds with half-rate LV applications.

A new radar determination of the spin vector of Venus

NASA Technical Reports Server (NTRS)

Zohar, S.; Goldstein, R. M.; Rumsey, H. C.

1980-01-01

Two radar observations of a set of three relatively small features on the surface of Venus have facilitated a refined determination of the spin vector of Venus. The period is found to be 243.019 + or 0.014 days, while the obliquity is 177.22 + or - 0.18 deg. The effects of deviations from exact sphericity on the interpretation of the measurements are discussed at length and the question of resonance with earth is reexamined.

Development of Peritoneal Tumor-Targeting Vector by In Vivo Screening with a Random Peptide-Displaying Adenovirus Library

PubMed Central

Yoshida, Kimiko; Goto, Naoko; Ohnami, Shumpei; Aoki, Kazunori

2012-01-01

The targeting of gene transfer at the cell-entry level is one of the most attractive challenges in vector development. However, attempts to redirect adenovirus vectors to alternative receptors by engineering the capsid-coding region have shown limited success, because the proper targeting ligands on the cells of interest are generally unknown. To overcome this limitation, we have constructed a random peptide library displayed on the adenoviral fiber knob, and have successfully selected targeted vectors by screening the library on cancer cell lines in vitro. The infection of targeted vectors was considered to be mediated by specific receptors on target cells. However, the expression levels and kinds of cell surface receptors may be substantially different between in vitro culture and in vivo tumor tissue. Here, we screened the peptide display-adenovirus library in the peritoneal dissemination model of AsPC-1 pancreatic cancer cells. The vector displaying a selected peptide (PFWSGAV) showed higher infectivity in the AsPC-1 peritoneal tumors but not in organs and other peritoneal tumors as compared with a non-targeted vector. Furthermore, the infectivity of the PFWSGAV-displaying vector for AsPC-1 peritoneal tumors was significantly higher than that of a vector displaying a peptide selected by in vitro screening, indicating the usefulness of in vivo screening in exploring the targeting vectors. This vector-screening system can facilitate the development of targeted adenovirus vectors for a variety of applications in medicine. PMID:23029088

GenoCAD Plant Grammar to Design Plant Expression Vectors for Promoter Analysis.

PubMed

Coll, Anna; Wilson, Mandy L; Gruden, Kristina; Peccoud, Jean

2016-01-01

With the rapid advances in prediction tools for discovery of new promoters and their cis-elements, there is a need to improve plant expression methodologies in order to facilitate a high-throughput functional validation of these promoters in planta. The promoter-reporter analysis is an indispensible approach for characterization of plant promoters. It requires the design of complex plant expression vectors, which can be challenging. Here, we describe the use of a plant grammar implemented in GenoCAD that will allow the users to quickly design constructs for promoter analysis experiments but also for other in planta functional studies. The GenoCAD plant grammar includes a library of plant biological parts organized in structural categories to facilitate their use and management and a set of rules that guides the process of assembling these biological parts into large constructs.

Evaluating the promise of recombinant transmissible vaccines

PubMed Central

Basinski, Andrew J.; Varrelman, Tanner J.; Smithson, Mark W.; May, Ryan H.; Remien, Christopher H.; Nuismer, Scott L.

2018-01-01

Transmissible vaccines have the potential to revolutionize infectious disease control by reducing the vaccination effort required to protect a population against a disease. Recent efforts to develop transmissible vaccines focus on recombinant transmissible vaccine designs (RTVs) because they pose reduced risk if intra-host evolution causes the vaccine to revert to its vector form. However, the shared antigenicity of the vaccine and vector may confer vaccine-immunity to hosts infected with the vector, thwarting the ability of the vaccine to spread through the population. We build a mathematical model to test whether a RTV can facilitate disease management in instances where reversion is likely to introduce the vector into the population or when the vector organism is already established in the host population, and the vector and vaccine share perfect cross-immunity. Our results show that a RTV can autonomously eradicate a pathogen, or protect a population from pathogen invasion, when cross-immunity between vaccine and vector is absent. If cross-immunity between vaccine and vector exists, however, our results show that a RTV can substantially reduce the vaccination effort necessary to control or eradicate a pathogen only when continuously augmented with direct manual vaccination. These results demonstrate that estimating the extent of cross-immunity between vector and vaccine is a critical step in RTV design, and that herpesvirus vectors showing facile reinfection and weak cross-immunity are promising. PMID:29279283

Transcriptome of the adult female malaria mosquito vector Anopheles albimanus.

PubMed

Martínez-Barnetche, Jesús; Gómez-Barreto, Rosa E; Ovilla-Muñoz, Marbella; Téllez-Sosa, Juan; García López, David E; Dinglasan, Rhoel R; Ubaida Mohien, Ceereena; MacCallum, Robert M; Redmond, Seth N; Gibbons, John G; Rokas, Antonis; Machado, Carlos A; Cazares-Raga, Febe E; González-Cerón, Lilia; Hernández-Martínez, Salvador; Rodríguez López, Mario H

2012-05-30

Human Malaria is transmitted by mosquitoes of the genus Anopheles. Transmission is a complex phenomenon involving biological and environmental factors of humans, parasites and mosquitoes. Among more than 500 anopheline species, only a few species from different branches of the mosquito evolutionary tree transmit malaria, suggesting that their vectorial capacity has evolved independently. Anopheles albimanus (subgenus Nyssorhynchus) is an important malaria vector in the Americas. The divergence time between Anopheles gambiae, the main malaria vector in Africa, and the Neotropical vectors has been estimated to be 100 My. To better understand the biological basis of malaria transmission and to develop novel and effective means of vector control, there is a need to explore the mosquito biology beyond the An. gambiae complex. We sequenced the transcriptome of the An. albimanus adult female. By combining Sanger, 454 and Illumina sequences from cDNA libraries derived from the midgut, cuticular fat body, dorsal vessel, salivary gland and whole body, we generated a single, high-quality assembly containing 16,669 transcripts, 92% of which mapped to the An. darlingi genome and covered 90% of the core eukaryotic genome. Bidirectional comparisons between the An. gambiae, An. darlingi and An. albimanus predicted proteomes allowed the identification of 3,772 putative orthologs. More than half of the transcripts had a match to proteins in other insect vectors and had an InterPro annotation. We identified several protein families that may be relevant to the study of Plasmodium-mosquito interaction. An open source transcript annotation browser called GDAV (Genome-Delinked Annotation Viewer) was developed to facilitate public access to the data generated by this and future transcriptome projects. We have explored the adult female transcriptome of one important New World malaria vector, An. albimanus. We identified protein-coding transcripts involved in biological processes that may be relevant to the Plasmodium lifecycle and can serve as the starting point for searching targets for novel control strategies. Our data increase the available genomic information regarding An. albimanus several hundred-fold, and will facilitate molecular research in medical entomology, evolutionary biology, genomics and proteomics of anopheline mosquito vectors. The data reported in this manuscript is accessible to the community via the VectorBase website (http://www.vectorbase.org/Other/AdditionalOrganisms/).

Surface modification via strain-promoted click reaction facilitates targeted lentiviral transduction.

PubMed

Chu, Yanjie; Oum, Yoon Hyeun; Carrico, Isaac S

2016-01-01

As a result of their ability to integrate into the genome of both dividing and non-dividing cells, lentiviruses have emerged as a promising vector for gene delivery. Targeted gene transduction of specific cells and tissues by lentiviral vectors has been a major goal, which has proven difficult to achieve. We report a novel targeting protocol that relies on the chemoselective attachment of cancer specific ligands to unnatural glycans on lentiviral surfaces. This strategy exhibits minimal perturbation on virus physiology and demonstrates remarkable flexibility. It allows for targeting but can be more broadly useful with applications such as vector purification and immunomodulation. Copyright © 2015 Elsevier Inc. All rights reserved.

Genomic approaches for understanding dengue: insights from the virus, vector, and host.

PubMed

Sim, Shuzhen; Hibberd, Martin L

2016-03-02

The incidence and geographic range of dengue have increased dramatically in recent decades. Climate change, rapid urbanization and increased global travel have facilitated the spread of both efficient mosquito vectors and the four dengue virus serotypes between population centers. At the same time, significant advances in genomics approaches have provided insights into host-pathogen interactions, immunogenetics, and viral evolution in both humans and mosquitoes. Here, we review these advances and the innovative treatment and control strategies that they are inspiring.

Compositional and phase relations among rare earth element minerals

NASA Technical Reports Server (NTRS)

Burt, D. M.

1990-01-01

This paper discusses the compositional and phase relationships among minerals in which rare earth elements (REE) occur as essential constituents (e.g., bastnaesite, monazite, xenotime, aeschynite, allanite). Particular consideration is given to the vector representation of complex coupled substitutions in selected REE-bearing minerals and to the REE partitioning between minerals as related to the acid-base tendencies and mineral stabilities. It is shown that the treatment of coupled substitutions as vector quantities facilitates graphical representation of mineral composition spaces.

Generation of arbitrary vector beams with liquid crystal polarization converters and vector-photoaligned q-plates

DOE Office of Scientific and Technical Information (OSTI.GOV)

Chen, Peng; Ji, Wei; Wei, Bing-Yan

Arbitrary vector beams (VBs) are realized by the designed polarization converters and corresponding vector-photoaligned q-plates. The polarization converter is a specific twisted nematic cell with one substrate homogeneously aligned and the other space-variantly aligned. By combining a polarization-sensitive alignment agent with a dynamic micro-lithography system, various categories of liquid crystal polarization converters are demonstrated. Besides, traditional radially/azimuthally polarized light, high-order and multi-ringed VBs, and a VB array with different orders are generated. The obtained converters are further utilized as polarization masks to implement vector-photoaligning. The technique facilitates both the volume duplication of these converters and the generation of another promisingmore » optical element, the q-plate, which is suitable for the generation of VBs for coherent lasers. The combination of proposed polarization converters and correspondingly fabricated q-plates would drastically enhance the capability of polarization control and may bring more possibilities for the design of photonic devices.« less

Automated solid-phase subcloning based on beads brought into proximity by magnetic force.

PubMed

Hudson, Elton P; Nikoshkov, Andrej; Uhlen, Mathias; Rockberg, Johan

2012-01-01

In the fields of proteomics, metabolic engineering and synthetic biology there is a need for high-throughput and reliable cloning methods to facilitate construction of expression vectors and genetic pathways. Here, we describe a new approach for solid-phase cloning in which both the vector and the gene are immobilized to separate paramagnetic beads and brought into proximity by magnetic force. Ligation events were directly evaluated using fluorescent-based microscopy and flow cytometry. The highest ligation efficiencies were obtained when gene- and vector-coated beads were brought into close contact by application of a magnet during the ligation step. An automated procedure was developed using a laboratory workstation to transfer genes into various expression vectors and more than 95% correct clones were obtained in a number of various applications. The method presented here is suitable for efficient subcloning in an automated manner to rapidly generate a large number of gene constructs in various vectors intended for high throughput applications.

Automated Solid-Phase Subcloning Based on Beads Brought into Proximity by Magnetic Force

PubMed Central

Hudson, Elton P.; Nikoshkov, Andrej; Uhlen, Mathias; Rockberg, Johan

2012-01-01

In the fields of proteomics, metabolic engineering and synthetic biology there is a need for high-throughput and reliable cloning methods to facilitate construction of expression vectors and genetic pathways. Here, we describe a new approach for solid-phase cloning in which both the vector and the gene are immobilized to separate paramagnetic beads and brought into proximity by magnetic force. Ligation events were directly evaluated using fluorescent-based microscopy and flow cytometry. The highest ligation efficiencies were obtained when gene- and vector-coated beads were brought into close contact by application of a magnet during the ligation step. An automated procedure was developed using a laboratory workstation to transfer genes into various expression vectors and more than 95% correct clones were obtained in a number of various applications. The method presented here is suitable for efficient subcloning in an automated manner to rapidly generate a large number of gene constructs in various vectors intended for high throughput applications. PMID:22624028

Development of Sendai Virus Vectors and their Potential Applications in Gene Therapy and Regenerative Medicine

PubMed Central

Nakanishi, Mahito; Otsu, Makoto

2012-01-01

Gene delivery/expression vectors have been used as fundamental technologies in gene therapy since the 1980s. These technologies are also being applied in regenerative medicine as tools to reprogram cell genomes to a pluripotent state and to other cell lineages. Rapid progress in these new research areas and expectations for their translation into clinical applications have facilitated the development of more sophisticated gene delivery/expression technologies. Since its isolation in 1953 in Japan, Sendai virus (SeV) has been widely used as a research tool in cell biology and in industry, but the application of SeV as a recombinant viral vector has been investigated only recently. Recombinant SeV vectors have various unique characteristics, such as low pathogenicity, powerful capacity for gene expression and a wide host range. In addition, the cytoplasmic gene expression mediated by this vector is advantageous for applications, in that chromosomal integration of exogenous genes can be undesirable. In this review, we introduce a brief historical background on the development of recombinant SeV vectors and describe their current applications in gene therapy. We also describe the application of SeV vectors in advanced nuclear reprogramming and introduce a defective and persistent SeV vector (SeVdp) optimized for such reprogramming. PMID:22920683

World of intelligence defense object detection-machine learning (artificial intelligence)

NASA Astrophysics Data System (ADS)

Gupta, Anitya; Kumar, Akhilesh; Bhushan, Vinayak

2018-04-01

This paper proposes a Quick Locale based Convolutional System strategy (Quick R-CNN) for question recognition. Quick R-CNN expands on past work to effectively characterize ob-ject recommendations utilizing profound convolutional systems. Com-pared to past work, Quick R-CNN utilizes a few in-novations to enhance preparing and testing speed while likewise expanding identification precision. Quick R-CNN trains the profound VGG16 arrange 9 quicker than R-CNN, is 213 speedier at test-time, and accomplishes a higher Guide on PASCAL VOC 2012. Contrasted with SPPnet, Quick R-CNN trains VGG16 3 quicker, tests 10 speedier, and is more exact. Quick R-CNN is actualized in Python and C++ (utilizing Caffe) and is accessible under the open-source MIT Permit.

The ecological foundations of transmission potential and vector-borne disease in urban landscapes.

PubMed

LaDeau, Shannon L; Allan, Brian F; Leisnham, Paul T; Levy, Michael Z

2015-07-01

Urban transmission of arthropod-vectored disease has increased in recent decades. Understanding and managing transmission potential in urban landscapes requires integration of sociological and ecological processes that regulate vector population dynamics, feeding behavior, and vector-pathogen interactions in these unique ecosystems. Vectorial capacity is a key metric for generating predictive understanding about transmission potential in systems with obligate vector transmission. This review evaluates how urban conditions, specifically habitat suitability and local temperature regimes, and the heterogeneity of urban landscapes can influence the biologically-relevant parameters that define vectorial capacity: vector density, survivorship, biting rate, extrinsic incubation period, and vector competence.Urban landscapes represent unique mosaics of habitat. Incidence of vector-borne disease in urban host populations is rarely, if ever, evenly distributed across an urban area. The persistence and quality of vector habitat can vary significantly across socio-economic boundaries to influence vector species composition and abundance, often generating socio-economically distinct gradients of transmission potential across neighborhoods.Urban regions often experience unique temperature regimes, broadly termed urban heat islands (UHI). Arthropod vectors are ectothermic organisms and their growth, survival, and behavior are highly sensitive to environmental temperatures. Vector response to UHI conditions is dependent on regional temperature profiles relative to the vector's thermal performance range. In temperate climates UHI can facilitate increased vector development rates while having countervailing influence on survival and feeding behavior. Understanding how urban heat island (UHI) conditions alter thermal and moisture constraints across the vector life cycle to influence transmission processes is an important direction for both empirical and modeling research.There remain persistent gaps in understanding of vital rates and drivers in mosquito-vectored disease systems, and vast holes in understanding for other arthropod vectored diseases. Empirical studies are needed to better understand the physiological constraints and socio-ecological processes that generate heterogeneity in critical transmission parameters, including vector survival and fitness. Likewise, laboratory experiments and transmission models must evaluate vector response to realistic field conditions, including variability in sociological and environmental conditions.

Production of SV40-derived vectors.

PubMed

Strayer, David S; Mitchell, Christine; Maier, Dawn A; Nichols, Carmen N

2010-06-01

Recombinant simian virus 40 (rSV40)-derived vectors are particularly useful for gene delivery to bone marrow progenitor cells and their differentiated derivatives, certain types of epithelial cells (e.g., hepatocytes), and central nervous system neurons and microglia. They integrate rapidly into cellular DNA to provide long-term gene expression in vitro and in vivo in both resting and dividing cells. Here we describe a protocol for production and purification of these vectors. These procedures require only packaging cells (e.g., COS-7) and circular vector genome DNA. Amplification involves repeated infection of packaging cells with vector produced by transfection. Cotransfection is not required in any step. Viruses are purified by centrifugation using discontinuous sucrose or cesium chloride (CsCl) gradients and resulting vectors are replication-incompetent and contain no detectable wild-type SV40 revertants. These approaches are simple, give reproducible results, and may be used to generate vectors that are deleted only for large T antigen (Tag), or for all SV40-coding sequences capable of carrying up to 5 kb of foreign DNA. These vectors are best applied to long-term expression of proteins normally encoded by mammalian cells or by viruses that infect mammalian cells, or of untranslated RNAs (e.g., RNA interference). The preparative approaches described facilitate application of these vectors and allow almost any laboratory to exploit their strengths for diverse gene delivery applications.

Bioinspired Star-Shaped Poly(l-lysine) Polypeptides: Efficient Polymeric Nanocarriers for the Delivery of DNA to Mesenchymal Stem Cells.

PubMed

Walsh, David P; Murphy, Robert D; Panarella, Angela; Raftery, Rosanne M; Cavanagh, Brenton; Simpson, Jeremy C; O'Brien, Fergal J; Heise, Andreas; Cryan, Sally-Ann

2018-05-07

The field of tissue engineering is increasingly recognizing that gene therapy can be employed for modulating in vivo cellular response thereby guiding tissue regeneration. However, the field lacks a versatile and biocompatible gene delivery platform capable of efficiently delivering transgenes to mesenchymal stem cells (MSCs), a cell type often refractory to transfection. Herein, we describe the extensive and systematic exploration of three architectural variations of star-shaped poly(l-lysine) polypeptide (star-PLL) with varying number and length of poly(l-lysine) arms as potential nonviral gene delivery vectors for MSCs. We demonstrate that star-PLL vectors are capable of self-assembling with pDNA to form stable, cationic nanomedicines. Utilizing high content screening, live cell imaging, and mechanistic uptake studies we confirm the intracellular delivery of pDNA by star-PLLs to MSCs is a rapid process, which likely proceeds via a clathrin-independent mechanism. We identify a star-PLL composition with 64 poly(l-lysine) arms and five l-lysine subunits per arm as a particularly efficient vector that is capable of delivering both reporter genes and the therapeutic transgenes bone morphogenetic protein-2 and vascular endothelial growth factor to MSCs. This composition facilitated a 1000-fold increase in transgene expression in MSCs compared to its linear analogue, linear poly(l-lysine). Furthermore, it demonstrated comparable transgene expression to the widely used vector polyethylenimine using a lower pDNA dose with significantly less cytotoxicity. Overall, this study illustrates the ability of the star-PLL vectors to facilitate efficient, nontoxic nucleic acid delivery to MSCs thereby functioning as an innovative nanomedicine platform for tissue engineering applications.

Exploring the role of peptides in polymer-based gene delivery.

PubMed

Sun, Yanping; Yang, Zhen; Wang, Chunxi; Yang, Tianzhi; Cai, Cuifang; Zhao, Xiaoyun; Yang, Li; Ding, Pingtian

2017-09-15

Polymers are widely studied as non-viral gene vectors because of their strong DNA binding ability, capacity to carry large payload, flexibility of chemical modifications, low immunogenicity, and facile processes for manufacturing. However, high cytotoxicity and low transfection efficiency substantially restrict their application in clinical trials. Incorporating functional peptides is a promising approach to address these issues. Peptides demonstrate various functions in polymer-based gene delivery systems, such as targeting to specific cells, breaching membrane barriers, facilitating DNA condensation and release, and lowering cytotoxicity. In this review, we systematically summarize the role of peptides in polymer-based gene delivery, and elaborate how to rationally design polymer-peptide based gene delivery vectors. Polymers are widely studied as non-viral gene vectors, but suffer from high cytotoxicity and low transfection efficiency. Incorporating short, bioactive peptides into polymer-based gene delivery systems can address this issue. Peptides demonstrate various functions in polymer-based gene delivery systems, such as targeting to specific cells, breaching membrane barriers, facilitating DNA condensation and release, and lowering cytotoxicity. In this review, we highlight the peptides' roles in polymer-based gene delivery, and elaborate how to utilize various functional peptides to enhance the transfection efficiency of polymers. The optimized peptide-polymer vectors should be able to alter their structures and functions according to biological microenvironments and utilize inherent intracellular pathways of cells, and consequently overcome the barriers during gene delivery to enhance transfection efficiency. Copyright © 2017 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Vectors, hosts, and control measures for Zika virus in the Americas

USGS Publications Warehouse

Thompson, Sarah J.; Pearce, John; Ramey, Andy M.

2017-01-01

We examine Zika virus (ZIKV) from an ecological perspective and with a focus on the Americas. We assess (1) the role of wildlife in ZIKV disease ecology, (2) how mosquito behavior and biology influence disease dynamics, and (3) how nontarget species and ecosystems may be impacted by vector control programs. Our review suggests that free-ranging, non-human primates may be involved in ZIKV transmission in the Old World; however, other wildlife species likely play a limited role in maintaining or transmitting ZIKV. In the Americas, a zoonotic cycle has not yet been definitively established. Understanding behaviors and habitat tolerances of Aedes aegypti and Aedes albopictus, two ZIKV competent vectors in the Americas, will allow more accurate modeling of disease spread and facilitate targeted and effective control efforts. Vector control efforts may have direct and indirect impacts to wildlife, particularly invertebrate feeding species; however, strategies could be implemented to limit detrimental ecological effects.

The Role of Bacterial Chaperones in the Circulative Transmission of Plant Viruses by Insect Vectors

PubMed Central

Kliot, Adi; Ghanim, Murad

2013-01-01

Persistent circulative transmission of plant viruses involves complex interactions between the transmitted virus and its insect vector. Several studies have shown that insect vector proteins are involved in the passage and the transmission of the virus. Interestingly, proteins expressed by bacterial endosymbionts that reside in the insect vector, were also shown to influence the transmission of these viruses. Thus far, the transmission of two plant viruses that belong to different virus genera was shown to be facilitated by a bacterial chaperone protein called GroEL. This protein was shown to be implicated in the transmission of Potato leafroll virus (PLRV) by the green peach aphid Myzus persicae, and the transmission of Tomato yellow leaf curl virus (TYLCV) by the sweetpotato whitefly Bemisia tabaci. These tri-trophic levels of interactions and their possible evolutionary implications are reviewed. PMID:23783810

Patient perspectives on delays in diagnosis and treatment of cancer: a qualitative analysis of free-text data.

PubMed

Parsonage, Rachel K; Hiscock, Julia; Law, Rebecca-Jane; Neal, Richard D

2017-01-01

Earlier cancer diagnosis is crucial in improving cancer survival. The International Cancer Benchmarking Partnership Module 4 (ICBP4) is a quantitative survey study that explores the reasons for delays in diagnosis and treatment of breast, colorectal, lung, and ovarian cancer. To further understand the associated diagnostic processes, it is also important to explore the patient perspectives expressed in the free-text comments. To use the free-text data provided by patients completing the ICBP4 survey to augment the understanding of patients' perspectives of their diagnostic journey. Qualitative analysis of the free-text data collected in Wales between October 2013 and December 2014 as part of the ICBP4 survey. Newly-diagnosed patients with either breast, ovarian, colorectal, or lung cancer were identified from registry data and then invited by their GPs to participate in the survey. A thematic framework was used to analyse the free-text comments provided at the end of the ICBP4 survey. Of the 905 patients who returned a questionnaire, 530 included comments. The free-text data provided information about patients' perspectives of the diagnostic journey. Analysis identified factors that acted as either barriers or facilitators at different stages of the diagnostic process. Some factors, such as screening, doctor-patient familiarity, and private treatment, acted as both barriers and facilitators depending on the context. Factors identified in this study help to explain how existing models of cancer diagnosis (for example, the Pathways to Treatment Model) work in practice. It is important that clinicians are aware of how these factors may interact with individual clinical cases and either facilitate, or act as a barrier to, subsequent cancer diagnosis. Understanding and implementing this knowledge into clinical practice may result in quicker cancer diagnoses. © British Journal of General Practice 2017.

Hybrid Adeno-Associated Viral Vectors Utilizing Transposase-Mediated Somatic Integration for Stable Transgene Expression in Human Cells

PubMed Central

Zhang, Wenli; Solanki, Manish; Müther, Nadine; Ebel, Melanie; Wang, Jichang; Sun, Chuanbo; Izsvak, Zsuzsanna; Ehrhardt, Anja

2013-01-01

Recombinant adeno-associated viral (AAV) vectors have been shown to be one of the most promising vectors for therapeutic gene delivery because they can induce efficient and long-term transduction in non-dividing cells with negligible side-effects. However, as AAV vectors mostly remain episomal, vector genomes and transgene expression are lost in dividing cells. Therefore, to stably transduce cells, we developed a novel AAV/transposase hybrid-vector. To facilitate SB-mediated transposition from the rAAV genome, we established a system in which one AAV vector contains the transposon with the gene of interest and the second vector delivers the hyperactive Sleeping Beauty (SB) transposase SB100X. Human cells were infected with the AAV-transposon vector and the transposase was provided in trans either by transient and stable plasmid transfection or by AAV vector transduction. We found that groups which received the hyperactive transposase SB100X showed significantly increased colony forming numbers indicating enhanced integration efficiencies. Furthermore, we found that transgene copy numbers in transduced cells were dose-dependent and that predominantly SB transposase-mediated transposition contributed to stabilization of the transgene. Based on a plasmid rescue strategy and a linear-amplification mediated PCR (LAM-PCR) protocol we analysed the SB100X-mediated integration profile after transposition from the AAV vector. A total of 1840 integration events were identified which revealed a close to random integration profile. In summary, we show for the first time that AAV vectors can serve as template for SB transposase mediated somatic integration. We developed the first prototype of this hybrid-vector system which with further improvements may be explored for treatment of diseases which originate from rapidly dividing cells. PMID:24116154

Which green way: description of the intervention for mobilising against Aedes aegypti under difficult security conditions in southern Mexico.

PubMed

Morales-Perez, Arcadio; Nava-Aguilera, Elizabeth; Legorreta-Soberanis, José; Paredes-Solís, Sergio; Balanzar-Martínez, Alejandro; Serrano-de Los Santos, Felipe René; Ríos-Rivera, Claudia Erika; García-Leyva, Jaime; Ledogar, Robert J; Cockcroft, Anne; Andersson, Neil

2017-05-30

Community mobilisation for prevention requires engagement with and buy in from those communities. In the Mexico state of Guerrero, unprecedented social violence related to the narcotics trade has eroded most community structures. A recent randomised controlled trial in 90 coastal communities achieved sufficient mobilisation to reduce conventional vector density indicators, self-reported dengue illness and serologically proved dengue virus infection. The Camino Verde intervention was a participatory research protocol promoting local discussion of baseline evidence and co-design of vector control solutions. Training of facilitators emphasised community authorship rather than trying to convince communities to do specific activities. Several discussion groups in each intervention community generated a loose and evolving prevention plan. Facilitators trained brigadistas, the first wave of whom received a small monthly stipend. Increasing numbers of volunteers joined the effort without pay. All communities opted to work with schoolchildren and for house-to-house visits by brigadístas. Children joined the neighbourhood vector control movements where security conditions permitted. After 6 months, a peer evaluation involved brigadista visits between intervention communities to review and to share progress. Although most communities had no active social institutions at the outset, local action planning using survey data provided a starting point for community authorship. Well-known in their own communities, brigadistas faced little security risk compared with the facilitators who visited the communities, or with governmental programmes. We believe the training focus on evidence-based dialogue and a plural community ownership through multiple design groups were key to success under challenging security conditions. ISRCTN27581154 .

[Drug vectorization or how to modulate tissular and cellular distribution of biologically active compounds].

PubMed

Couvreur, P

2001-07-01

Drug vectorization has undergone considerable development over the last few years. This review focuses on the intravenous route of administration. Colloid formulations allow a modulation of drug tissue distribution. Using liposomes and nanoparticles with unmodified surfaces, drugs can be targeted to macrophages of the reticulum endothelium system. When the liposomes or nanoparticles are covered with hydrophilic or flexible polymers, the vascular phase can be favored in order, for example, to facilitate selective extravasation at a tumor site. Therapeutic applications of these systems are presented. The development of "intelligent" vectors capable of modulating intracellular distribution of an active compounds is an equally interesting approach, for example pH-sensitive liposomes or nanoparticles decorated with folic acid capable of targeting intracellular cytoplasm.

Electromagnetic fluctuation spectra of collective oscillations in magnetized Maxwellian equal mass plasmas for low-frequency waves

DOE Office of Scientific and Technical Information (OSTI.GOV)

Vafin, S.; Schlickeiser, R.; Yoon, P. H.

Recently, the general electromagnetic fluctuation theory for magnetized plasmas has been used to study the steady-state fluctuation spectra and the total intensity of low-frequency collective weakly damped modes for parallel wave vectors in Maxwellian plasmas. Now, we address the same question with respect to an arbitrary direction of the wave-vector. Here, we analyze this problem for equal mass plasmas. These plasmas are a very good tool to study various plasma phenomena, as they considerably facilitate the theoretical consideration and at the same time provide with their clear physical picture. Finally, we compare our results in the limiting case of parallelmore » wave vectors with the previous study.« less

Integrating an infectious disease programme into the primary health care service: a retrospective analysis of Chagas disease community-based surveillance in Honduras.

PubMed

Hashimoto, Ken; Zúniga, Concepción; Nakamura, Jiro; Hanada, Kyo

2015-03-24

Integration of disease-specific programmes into the primary health care (PHC) service has been attempted mostly in clinically oriented disease control such as HIV/AIDS and tuberculosis but rarely in vector control. Chagas disease is controlled principally by interventions against the triatomine vector. In Honduras, after successful reduction of household infestation by vertical approach, the Ministry of Health implemented community-based vector surveillance at the PHC services (health centres) to prevent the resurgence of infection. This paper retrospectively analyses the effects and process of integrating a Chagas disease vector surveillance system into health centres. We evaluated the effects of integration at six pilot sites in western Honduras during 2008-2011 on; surveillance performance; knowledge, attitude and practice in schoolchildren; reports of triatomine bug infestation and institutional response; and seroprevalence among children under 15 years of age. The process of integration of the surveillance system was analysed using the PRECEDE-PROCEED model for health programme planning. The model was employed to systematically determine influential and interactive factors which facilitated the integration process at different levels of the Ministry of Health and the community. Overall surveillance performance improved from 46 to 84 on a 100 point-scale. Schoolchildren's attitude (risk awareness) score significantly increased from 77 to 83 points. Seroprevalence declined from 3.4% to 0.4%. Health centres responded to the community bug reports by insecticide spraying. As key factors, the health centres had potential management capacity and influence over the inhabitants' behaviours and living environment directly and through community health volunteers. The National Chagas Programme played an essential role in facilitating changes with adequate distribution of responsibilities, participatory modelling, training and, evaluation and advocacy. We found that Chagas disease vector surveillance can be integrated into the PHC service. Health centres demonstrated capacity to manage vector surveillance and improve performance, children's awareness, vector report-response and seroprevalence, once tasks were simplified to be performed by trained non-specialists and distributed among the stakeholders. Health systems integration requires health workers to perform beyond their usual responsibilities and acquire management skills. Integration of vector control is feasible and can contribute to strengthening the preventive capacity of the PHC service.

Sever's Disease

MedlinePlus

... immobilize the foot so that it can heal. Recovery and Recurrence One of the most important things ... The sooner Sever's disease is addressed, the quicker recovery is. Most kids can return to physical activity ...

Genome Investigations of Vector Competence in Aedes aegypti to Inform Novel Arbovirus Disease Control Approaches

PubMed Central

Severson, David W.; Behura, Susanta K.

2016-01-01

Dengue (DENV), yellow fever, chikungunya, and Zika virus transmission to humans by a mosquito host is confounded by both intrinsic and extrinsic variables. Besides virulence factors of the individual arboviruses, likelihood of virus transmission is subject to variability in the genome of the primary mosquito vector, Aedes aegypti. The “vectorial capacity” of A. aegypti varies depending upon its density, biting rate, and survival rate, as well as its intrinsic ability to acquire, host and transmit a given arbovirus. This intrinsic ability is known as “vector competence”. Based on whole transcriptome analysis, several genes and pathways have been predicated to have an association with a susceptible or refractory response in A. aegypti to DENV infection. However, the functional genomics of vector competence of A. aegypti is not well understood, primarily due to lack of integrative approaches in genomic or transcriptomic studies. In this review, we focus on the present status of genomics studies of DENV vector competence in A. aegypti as limited information is available relative to the other arboviruses. We propose future areas of research needed to facilitate the integration of vector and virus genomics and environmental factors to work towards better understanding of vector competence and vectorial capacity in natural conditions. PMID:27809220

Tailor-made gene silencing of Staphylococcus aureus clinical isolates by CRISPR interference

PubMed Central

Sato’o, Yusuke; Hisatsune, Junzo; Yu, Liansheng; Sakuma, Tetsushi; Yamamoto, Takashi

2018-01-01

Preparing the genetically modified organisms have required much time and labor, making it the rate-limiting step but CRISPR/Cas9 technology appearance has changed this difficulty. Although reports on CRISPR/Cas9 technology such as genome editing and CRISPR interference (CRISPRi) in eukaryotes increased, those in prokaryotes especially in Staphylococci were limited. Thus, its potential in the bacteriology remains unexplored. This is attributed to ecological difference between eukaryotes and prokaryotes. Here, we constructed a novel CRISPRi plasmid vector, pBACi for Staphylococcus aureus. The transformation efficiency of S. aureus was ~104 CFU/μg DNA using a vector extracted from dcm negative, which encoded one of DNA modification genes, E. coli. Further, pBACi was introduced into various clinical isolates including that not accepting the conventional temperature-sensitive vector. dcas9 in the vector was expressed throughout the growth phases of S. aureus and this vector decreased various gene mRNA expressions based on the crRNA targeting sequences and altered the knockdown strains’ phenotypes. The targeted genes included various virulence and antibiotic resistant genes. Bioinformatics suggest this vector can be introduced into wide range of low-GC Gram-positive bacteria. Because this new CRISPR/Cas9-based vector can easily prepare knockdown strains, we believe the novel vector will facilitate the characterization of the function of genes from S. aureus and other Gram-positive bacteria. PMID:29377933

An Improved Single-Step Cloning Strategy Simplifies the Agrobacterium tumefaciens-Mediated Transformation (ATMT)-Based Gene-Disruption Method for Verticillium dahliae.

PubMed

Wang, Sheng; Xing, Haiying; Hua, Chenlei; Guo, Hui-Shan; Zhang, Jie

2016-06-01

The soilborne fungal pathogen Verticillium dahliae infects a broad range of plant species to cause severe diseases. The availability of Verticillium genome sequences has provided opportunities for large-scale investigations of individual gene function in Verticillium strains using Agrobacterium tumefaciens-mediated transformation (ATMT)-based gene-disruption strategies. Traditional ATMT vectors require multiple cloning steps and elaborate characterization procedures to achieve successful gene replacement; thus, these vectors are not suitable for high-throughput ATMT-based gene deletion. Several advancements have been made that either involve simplification of the steps required for gene-deletion vector construction or increase the efficiency of the technique for rapid recombinant characterization. However, an ATMT binary vector that is both simple and efficient is still lacking. Here, we generated a USER-ATMT dual-selection (DS) binary vector, which combines both the advantages of the USER single-step cloning technique and the efficiency of the herpes simplex virus thymidine kinase negative-selection marker. Highly efficient deletion of three different genes in V. dahliae using the USER-ATMT-DS vector enabled verification that this newly-generated vector not only facilitates the cloning process but also simplifies the subsequent identification of fungal homologous recombinants. The results suggest that the USER-ATMT-DS vector is applicable for efficient gene deletion and suitable for large-scale gene deletion in V. dahliae.

An integrated vector system for cellular studies of phage display-derived peptides.

PubMed

Voss, Stephan D; DeGrand, Alec M; Romeo, Giulio R; Cantley, Lewis C; Frangioni, John V

2002-09-15

Peptide phage display is a method by which large numbers of diverse peptides can be screened for binding to a target of interest. Even when successful, the rate-limiting step is usually validation of peptide bioactivity using living cells. In this paper, we describe an integrated system of vectors that expedites both the screening and the characterization processes. Library construction and screening is performed using an optimized type 3 phage display vector, mJ(1), which is shown to accept peptide libraries of at least 23 amino acids in length. Peptide coding sequences are shuttled from mJ(1) into one of three families of mammalian expression vectors for cell physiological studies. The vector pAL(1) expresses phage display-derived peptides as Gal4 DNA binding domain fusion proteins for transcriptional activation studies. The vectors pG(1), pG(1)N, and pG(1)C express phage display-derived peptides as green fluorescent protein fusions targeted to the entire cell, nucleus, or cytoplasm, respectively. The vector pAP(1) expresses phage display-derived peptides as fusions to secreted placental alkaline phosphatase. Such enzyme fusions can be used as highly sensitive affinity reagents for high-throughput assays and for cloning of peptide-binding cell surface receptors. Taken together, this system of vectors should facilitate the development of phage display-derived peptides into useful biomolecules.

Application of satellite data in variational analysis for global cyclonic systems

NASA Technical Reports Server (NTRS)

Achtemeier, G. L.

1987-01-01

The research goal was a variational data assimilation method that incorporates as dynamical constraints, the primitive equations for a moist, convectively unstable atmosphere and the radiative transfer equation. Variables to be adjusted include the three-dimensional vector wind, height, temperature, and moisture from rawinsonde data, and cloud-wind vectors, moisture, and radiance from satellite data. This presents a formidable mathematical problem. In order to facilitate thorough analysis of each of the model components, four variational models that divide the problem naturally according to increasing complexity are defined. Each model is summarized.

Enhanced green fluorescent protein (egfp) gene expression in Tetraselmis subcordiformis chloroplast with endogenous regulators.

PubMed

Cui, Yulin; Zhao, Jialin; Hou, Shichang; Qin, Song

2016-05-01

On the basis of fundamental genetic transformation technologies, the goal of this study was to optimize Tetraselmis subcordiformis chloroplast transformation through the use of endogenous regulators. The genes rrn16S, rbcL, psbA, and psbC are commonly highly expressed in chloroplasts, and the regulators of these genes are often used in chloroplast transformation. For lack of a known chloroplast genome sequence, the genome-walking method was used here to obtain full sequences of T. subcordiformis endogenous regulators. The resulting regulators, including three promoters, two terminators, and a ribosome combination sequence, were inserted into the previously constructed plasmid pPSC-R, with the egfp gene included as a reporter gene, and five chloroplast expression vectors prepared. These vectors were successfully transformed into T. subcordiformis by particle bombardment and the efficiency of each vector tested by assessing EGFP fluorescence via microscopy. The results showed that these vectors exhibited higher efficiency than the former vector pPSC-G carrying exogenous regulators, and the vector pRFA with Prrn, psbA-5'RE, and TpsbA showed the highest efficiency. This research provides a set of effective endogenous regulators for T. subcordiformis and will facilitate future fundamental studies of this alga.

Genome scaffolding and annotation for the pathogen vector Ixodes ricinus by ultra-long single molecule sequencing.

PubMed

Cramaro, Wibke J; Hunewald, Oliver E; Bell-Sakyi, Lesley; Muller, Claude P

2017-02-08

Global warming and other ecological changes have facilitated the expansion of Ixodes ricinus tick populations. Ixodes ricinus is the most important carrier of vector-borne pathogens in Europe, transmitting viruses, protozoa and bacteria, in particular Borrelia burgdorferi (sensu lato), the causative agent of Lyme borreliosis, the most prevalent vector-borne disease in humans in the Northern hemisphere. To faster control this disease vector, a better understanding of the I. ricinus tick is necessary. To facilitate such studies, we recently published the first reference genome of this highly prevalent pathogen vector. Here, we further extend these studies by scaffolding and annotating the first reference genome by using ultra-long sequencing reads from third generation single molecule sequencing. In addition, we present the first genome size estimation for I. ricinus ticks and the embryo-derived cell line IRE/CTVM19. 235,953 contigs were integrated into 204,904 scaffolds, extending the currently known genome lengths by more than 30% from 393 to 516 Mb and the N50 contig value by 87% from 1643 bp to a N50 scaffold value of 3067 bp. In addition, 25,263 sequences were annotated by comparison to the tick's North American relative Ixodes scapularis. After (conserved) hypothetical proteins, zinc finger proteins, secreted proteins and P450 coding proteins were the most prevalent protein categories annotated. Interestingly, more than 50% of the amino acid sequences matching the homology threshold had 95-100% identity to the corresponding I. scapularis gene models. The sequence information was complemented by the first genome size estimation for this species. Flow cytometry-based genome size analysis revealed a haploid genome size of 2.65Gb for I. ricinus ticks and 3.80 Gb for the cell line. We present a first draft sequence map of the I. ricinus genome based on a PacBio-Illumina assembly. The I. ricinus genome was shown to be 26% (500 Mb) larger than the genome of its American relative I. scapularis. Based on the genome size of 2.65 Gb we estimated that we covered about 67% of the non-repetitive sequences. Genome annotation will facilitate screening for specific molecular pathways in I. ricinus cells and provides an overview of characteristics and functions.

Taub-NUT Spacetime in the (A)dS/CFT and M-Theory [electronic resource

NASA Astrophysics Data System (ADS)

Clarkson, Richard

In the following thesis, I will conduct a thermodynamic analysis of the Taub-NUT spacetime in various dimensions, as well as show uses for Taub-NUT and other Hyper-Kahler spacetimes. Thermodynamic analysis (by which I mean the calculation of the entropy and other thermodynamic quantities, and the analysis of these quantities) has in the past been done by use of background subtraction. The recent derivation of the (A)dS/CFT correspondences from String theory has allowed for easier and quicker analysis. I will use Taub-NUT space as a template to test these correspondences against the standard thermodynamic calculations (via the N?ether method), with (in the Taub-NUT-dS case especially) some very interesting results. There is also interest in obtaining metrics in eleven dimensions that can be reduced down to ten dimensional string theory metrics. Taub-NUT and other Hyper-Kahler metrics already possess the form to easily facilitate the Kaluza-Klein reduction, and embedding such metricsinto eleven dimensional metrics containing M2 or M5 branes produces metrics with interesting Dp-brane results.

Memory updating in sub-clinical eating disorder: differential effects with food and body shape words.

PubMed

Fenton, Olivia; Ecker, Ullrich K H

2015-04-01

The present study investigated how eating disorder (ED) relevant information is updated in working memory in people with high vs. low scores on a measure of eating disorder pathology (the Eating Disorder Examination Questionnaire, EDE-Q). Participants performed two memory updating tasks. One was a neutral control task using digits; the other task involved food words and words relating to body-shape, and provided measures of updating speed and post-updating recall. We found that high EDE-Q participants (1) showed no sign of general memory updating impairment as indicated by performance in the control task; (2) showed a general recall deficit in the task involving ED-relevant stimuli, suggesting a general distraction of cognitive resources in the presence of ED-related items; (3) showed a relative facilitation in the recall of food words; and (4) showed quicker updating toward food words and relatively slower updating toward body-shape-related words. Results are discussed in the context of cognitive theories of eating disorders. Copyright © 2015 Elsevier Ltd. All rights reserved.

Single-incision laparoscopic distal gastrectomy for early gastric cancer through a homemade single port access device.

PubMed

Jiang, Zhi-Wei; Zhang, Shu; Wang, Gang; Zhao, Kun; Liu, Jiang; Ning, Li; Li, Jieshou

2015-01-01

We presented a series of single-incision laparoscopic distal gastrectomies for early gastric cancer patients through a type of homemade single port access device and some other conventional laparoscopic instruments. A single-incision laparoscopic distal gastrectomy with D1 + α lymph node dissection was performed on a 46 years old male patient who had an early gastric cancer. This single port access device has facilitated the conventional laparoscopic instruments to accomplish the surgery and we made in only 6 minutes. Total operating time for this surgery was 240 minutes. During the operation, there were about 100 milliliters of blood loss, and 17 lymph-nodes were retrieved. This homemade single port access device shows its superiority in economy and convenience for complex single-incision surgeries. Single-incision laparoscopic distal gastrectomy for early gastric cancer can be conducted by experienced laparoscopic surgeons. Fully take advantage of both SILS and fast track surgery plan can bring to successful surgeries with minimal postoperative pain, quicker mobilization, early recovery of intestinal function, and better cosmesis effect for the patients.

Amygdala activity for the modulation of goal-directed behavior in emotional contexts

PubMed Central

Kunimatsu, Jun; Hikosaka, Okihide

2018-01-01

Choosing valuable objects and rewarding actions is critical for survival. While such choices must be made in a way that suits the animal’s circumstances, the neural mechanisms underlying such context-appropriate behavior are unclear. To address this question, we devised a context-dependent reward-seeking task for macaque monkeys. Each trial started with the appearance of one of many visual scenes containing two or more objects, and the monkey had to choose the good object by saccade to get a reward. These scenes were categorized into two dimensions of emotional context: dangerous versus safe and rich versus poor. We found that many amygdala neurons were more strongly activated by dangerous scenes, by rich scenes, or by both. Furthermore, saccades to target objects occurred more quickly in dangerous than in safe scenes and were also quicker in rich than in poor scenes. Thus, amygdala neuronal activity and saccadic reaction times were negatively correlated in each monkey. These results suggest that amygdala neurons facilitate targeting saccades predictably based on aspects of emotional context, as is necessary for goal-directed and social behavior. PMID:29870524

Plant Virus–Insect Vector Interactions: Current and Potential Future Research Directions

PubMed Central

Dietzgen, Ralf G.; Mann, Krin S.; Johnson, Karyn N.

2016-01-01

Acquisition and transmission by an insect vector is central to the infection cycle of the majority of plant pathogenic viruses. Plant viruses can interact with their insect host in a variety of ways including both non-persistent and circulative transmission; in some cases, the latter involves virus replication in cells of the insect host. Replicating viruses can also elicit both innate and specific defense responses in the insect host. A consistent feature is that the interaction of the virus with its insect host/vector requires specific molecular interactions between virus and host, commonly via proteins. Understanding the interactions between plant viruses and their insect host can underpin approaches to protect plants from infection by interfering with virus uptake and transmission. Here, we provide a perspective focused on identifying novel approaches and research directions to facilitate control of plant viruses by better understanding and targeting virus–insect molecular interactions. We also draw parallels with molecular interactions in insect vectors of animal viruses, and consider technical advances for their control that may be more broadly applicable to plant virus vectors. PMID:27834855

Plant Virus-Insect Vector Interactions: Current and Potential Future Research Directions.

PubMed

Dietzgen, Ralf G; Mann, Krin S; Johnson, Karyn N

2016-11-09

Acquisition and transmission by an insect vector is central to the infection cycle of the majority of plant pathogenic viruses. Plant viruses can interact with their insect host in a variety of ways including both non-persistent and circulative transmission; in some cases, the latter involves virus replication in cells of the insect host. Replicating viruses can also elicit both innate and specific defense responses in the insect host. A consistent feature is that the interaction of the virus with its insect host/vector requires specific molecular interactions between virus and host, commonly via proteins. Understanding the interactions between plant viruses and their insect host can underpin approaches to protect plants from infection by interfering with virus uptake and transmission. Here, we provide a perspective focused on identifying novel approaches and research directions to facilitate control of plant viruses by better understanding and targeting virus-insect molecular interactions. We also draw parallels with molecular interactions in insect vectors of animal viruses, and consider technical advances for their control that may be more broadly applicable to plant virus vectors.

Vectorcardiographic diagnostic & prognostic information derived from the 12-lead electrocardiogram: Historical review and clinical perspective.

PubMed

Man, Sumche; Maan, Arie C; Schalij, Martin J; Swenne, Cees A

2015-01-01

In the course of time, electrocardiography has assumed several modalities with varying electrode numbers, electrode positions and lead systems. 12-lead electrocardiography and 3-lead vectorcardiography have become particularly popular. These modalities developed in parallel through the mid-twentieth century. In the same time interval, the physical concepts underlying electrocardiography were defined and worked out. In particular, the vector concept (heart vector, lead vector, volume conductor) appeared to be essential to understanding the manifestations of electrical heart activity, both in the 12-lead electrocardiogram (ECG) and in the 3-lead vectorcardiogram (VCG). Not universally appreciated in the clinic, the vectorcardiogram, and with it the vector concept, went out of use. A revival of vectorcardiography started in the 90's, when VCGs were mathematically synthesized from standard 12-lead ECGs. This facilitated combined electrocardiography and vectorcardiography without the need for a special recording system. This paper gives an overview of these historical developments, elaborates on the vector concept and seeks to define where VCG analysis/interpretation can add diagnostic/prognostic value to conventional 12-lead ECG analysis. Copyright © 2015 Elsevier Inc. All rights reserved.

Kidney removal

MedlinePlus

... the surgical cut is located. Recovery after a laparoscopic procedure is most often quicker, with less pain. Outlook (Prognosis) The outcome is most often good when a single kidney is removed. If both kidneys are removed, ...

A multi-dimensional Smolyak collocation method in curvilinear coordinates for computing vibrational spectra

DOE Office of Scientific and Technical Information (OSTI.GOV)

Avila, Gustavo, E-mail: Gustavo-Avila@telefonica.net; Carrington, Tucker, E-mail: Tucker.Carrington@queensu.ca

In this paper, we improve the collocation method for computing vibrational spectra that was presented in Avila and Carrington, Jr. [J. Chem. Phys. 139, 134114 (2013)]. Using an iterative eigensolver, energy levels and wavefunctions are determined from values of the potential on a Smolyak grid. The kinetic energy matrix-vector product is evaluated by transforming a vector labelled with (nondirect product) grid indices to a vector labelled by (nondirect product) basis indices. Both the transformation and application of the kinetic energy operator (KEO) scale favorably. Collocation facilitates dealing with complicated KEOs because it obviates the need to calculate integrals of coordinatemore » dependent coefficients of differential operators. The ideas are tested by computing energy levels of HONO using a KEO in bond coordinates.« less

Co-feeding transmission facilitates strain coexistence in Borrelia burgdorferi, the Lyme disease agent.

PubMed

States, S L; Huang, C I; Davis, S; Tufts, D M; Diuk-Wasser, M A

2017-06-01

Coexistence of multiple tick-borne pathogens or strains is common in natural hosts and can be facilitated by resource partitioning of the host species, within-host localization, or by different transmission pathways. Most vector-borne pathogens are transmitted horizontally via systemic host infection, but transmission may occur in the absence of systemic infection between two vectors feeding in close proximity, enabling pathogens to minimize competition and escape the host immune response. In a laboratory study, we demonstrated that co-feeding transmission can occur for a rapidly-cleared strain of Borrelia burgdorferi, the Lyme disease agent, between two stages of the tick vector Ixodes scapularis while feeding on their dominant host, Peromyscus leucopus. In contrast, infections rapidly became systemic for the persistently infecting strain. In a field study, we assessed opportunities for co-feeding transmission by measuring co-occurrence of two tick stages on ears of small mammals over two years at multiple sites. Finally, in a modeling study, we assessed the importance of co-feeding on R 0 , the basic reproductive number. The model indicated that co-feeding increases the fitness of rapidly-cleared strains in regions with synchronous immature tick feeding. Our results are consistent with increased diversity of B. burgdorferi in areas of higher synchrony in immature feeding - such as the midwestern United States. A higher relative proportion of rapidly-cleared strains, which are less human pathogenic, would also explain lower Lyme disease incidence in this region. Finally, if co-feeding transmission also occurs on refractory hosts, it may facilitate the emergence and persistence of new pathogens with a more limited host range. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

A superhydrophobic cone to facilitate the xenomonitoring of filarial parasites, malaria, and trypanosomes using mosquito excreta/feces.

PubMed

Cook, Darren A N; Pilotte, Nils; Minetti, Corrado; Williams, Steven A; Reimer, Lisa J

2017-11-06

Background: Molecular xenomonitoring (MX), the testing of insect vectors for the presence of human pathogens, has the potential to provide a non-invasive and cost-effective method for monitoring the prevalence of disease within a community. Current MX methods require the capture and processing of large numbers of mosquitoes, particularly in areas of low endemicity, increasing the time, cost and labour required. Screening the excreta/feces (E/F) released from mosquitoes, rather than whole carcasses, improves the throughput by removing the need to discriminate vector species since non-vectors release ingested pathogens in E/F. It also enables larger numbers of mosquitoes to be processed per pool. However, this new screening approach requires a method of efficiently collecting E/F. Methods: We developed a cone with a superhydrophobic surface to allow for the efficient collection of E/F. Using mosquitoes exposed to either Plasmodium falciparum , Brugia malayi or Trypanosoma brucei brucei, we tested the performance of the superhydrophobic cone alongside two other collection methods. Results: All collection methods enabled the detection of DNA from the three parasites. Using the superhydrophobic cone to deposit E/F into a small tube provided the highest number of positive samples (16 out of 18) and facilitated detection of parasite DNA in E/F from individual mosquitoes. Further tests showed that following a simple washing step, the cone can be reused multiple times, further improving its cost-effectiveness. Conclusions: Incorporating the superhydrophobic cone into mosquito traps or holding containers could provide a simple and efficient method for collecting E/F. Where this is not possible, swabbing the container or using the washing method facilitates the detection of the three parasites used in this study.

Quicker Veterans Benefits Delivery Act of 2013

THOMAS, 113th Congress

Sen. Franken, Al [D-MN

2013-05-13

Senate - 06/12/2013 Committee on Veterans' Affairs. Hearings held. Hearings printed: S.Hrg. 113-111. (All Actions) Tracker: This bill has the status IntroducedHere are the steps for Status of Legislation:

The pandemonium system of reflective agents.

PubMed

Smieja, F

1996-01-01

The Pandemonium system of reflective MINOS agents solves problems by automatic dynamic modularization of the input space. The agents contain feedforward neural networks which adapt using the backpropagation algorithm. We demonstrate the performance of Pandemonium on various categories of problems. These include learning continuous functions with discontinuities, separating two spirals, learning the parity function, and optical character recognition. It is shown how strongly the advantages gained from using a modularization technique depend on the nature of the problem. The superiority of the Pandemonium method over a single net on the first two test categories is contrasted with its limited advantages for the second two categories. In the first case the system converges quicker with modularization and is seen to lead to simpler solutions. For the second case the problem is not significantly simplified through flat decomposition of the input space, although convergence is still quicker.

Circadian rhythms in insect disease vectors

PubMed Central

Meireles-Filho, Antonio Carlos Alves; Kyriacou, Charalambos Panayiotis

2013-01-01

Organisms from bacteria to humans have evolved under predictable daily environmental cycles owing to the Earth’s rotation. This strong selection pressure has generated endogenous circadian clocks that regulate many aspects of behaviour, physiology and metabolism, anticipating and synchronising internal time-keeping to changes in the cyclical environment. In haematophagous insect vectors the circadian clock coordinates feeding activity, which is important for the dynamics of pathogen transmission. We have recently witnessed a substantial advance in molecular studies of circadian clocks in insect vector species that has consolidated behavioural data collected over many years, which provided insights into the regulation of the clock in the wild. Next generation sequencing technologies will facilitate the study of vector genomes/transcriptomes both among and within species and illuminate some of the species-specific patterns of adaptive circadian phenotypes that are observed in the field and in the laboratory. In this review we will explore these recent findings and attempt to identify potential areas for further investigation. PMID:24473802

Pullulan-protamine as efficient haemocompatible gene delivery vector: synthesis and in vitro characterization.

PubMed

Priya, S S; Rekha, M R; Sharma, Chandra P

2014-02-15

Biodegradable non-viral vectors with good transfection efficiency is essential for successful gene delivery. The purpose of this study was to design a non-viral vector by conjugating protamine to pullulan and elucidate the potential use of pullulan protamine conjugate (PPA) as an effective, non toxic and haemocompatible gene delivery system. The particle size and surface charge were measured using Nanosizer. Derivatization was confirmed by NMR, FTIR and DSC analyses. Acid base titration revealed the buffering behaviour of the conjugate. The protection of DNA from nuclease enzyme and interaction of plasma components on the stability of nanoplexes were also analysed. The uptake studies confirmed the plasmid delivery into the nucleus and the inhibitor studies determined the uptake mechanism. Transfection experiments revealed the capability of PPA to cellular uptake in C6 cells and facilitate high gene expression. Thus, PPA proves to be a promising non-viral vector. Copyright © 2013 Elsevier Ltd. All rights reserved.

Currentless reversal of Néel vector in antiferromagnets

NASA Astrophysics Data System (ADS)

Semenov, Yuriy; Li, Xilai; Kim, Ki Wook

The bias driven perpendicular magnetic anisotropy is a magneto-electric effect that can realize 900 magnetization rotation and even 1800 flip along the easy axis in the ferromagnets with a minimal energy consumption. This study theoretically demonstrates a similar phenomenon of the Néel vector reversal via a short electrical pulse that can mediate perpendicular magnetic anisotropy in the antiferromagnets. The analysis based on the dynamical equations as well as the micromagnetic simulations reveals the important role of the inertial behavior in the antiferromagnets that facilitates the Néel vector to overcome the barrier between two free-energy minima of the bistable states along the easy axis. In contrast to the ferromagnets, this Néel vector reversal does not accompany angular moment transfer to the environment, leading to acceleration in the dynamical response by a few orders of magnitude. Further, a small switching energy requirement of a few attojoules illustrates an added advantage of the phenomenon in low-power spintronic applications.

Plasimids containing the gene for DNA polymerase I from Streptococcus pneumoniae

DOEpatents

Lacks, Sanford A.; Martinez, Susana; Lopez, Paloma; Espinosa, Manuel

1991-01-01

A method is disclosed for cloning the gene which encodes a DNA polymerase-exonuclease of Streptococcus pneumoniae. Plasmid pSM22, the vector containing the pneumocccal polA gene, facilitates the expression of 50-fold greater amounts of the PolI enzyme.

The Birds and the Bees...and the Bats.

ERIC Educational Resources Information Center

Foote, MaryAnn

1990-01-01

Pollination vectors of a variety of types are described including beetles, bees, flies, moths, birds, bats, and the wind. Some of the adaptations of plants designed to help facilitate pollination are discussed. Strategies for incorporating this information into a lesson plan are suggested. (CW)

77 FR 67782 - Proposed Amendment of Class E Airspace; Casper, WY

Federal Register 2010, 2011, 2012, 2013, 2014

2012-11-14

..., Natrona County International Airport, Casper, WY, to facilitate vectoring of Instrument Flight Rules (IFR... Range Tactical Air Navigation (VORTAC) has made reconfiguration necessary for the safety and management of aircraft operations at Casper, Natrona County International Airport, Casper, WY. DATES: Comments...

Practical utilization of recombinant AAV vector reference standards: focus on vector genomes titration by free ITR qPCR.

PubMed

D'Costa, Susan; Blouin, Veronique; Broucque, Frederic; Penaud-Budloo, Magalie; François, Achille; Perez, Irene C; Le Bec, Christine; Moullier, Philippe; Snyder, Richard O; Ayuso, Eduard

2016-01-01

Clinical trials using recombinant adeno-associated virus (rAAV) vectors have demonstrated efficacy and a good safety profile. Although the field is advancing quickly, vector analytics and harmonization of dosage units are still a limitation for commercialization. AAV reference standard materials (RSMs) can help ensure product safety by controlling the consistency of assays used to characterize rAAV stocks. The most widely utilized unit of vector dosing is based on the encapsidated vector genome. Quantitative polymerase chain reaction (qPCR) is now the most common method to titer vector genomes (vg); however, significant inter- and intralaboratory variations have been documented using this technique. Here, RSMs and rAAV stocks were titered on the basis of an inverted terminal repeats (ITRs) sequence-specific qPCR and we found an artificial increase in vg titers using a widely utilized approach. The PCR error was introduced by using single-cut linearized plasmid as the standard curve. This bias was eliminated using plasmid standards linearized just outside the ITR region on each end to facilitate the melting of the palindromic ITR sequences during PCR. This new "Free-ITR" qPCR delivers vg titers that are consistent with titers obtained with transgene-specific qPCR and could be used to normalize in-house product-specific AAV vector standards and controls to the rAAV RSMs. The free-ITR method, including well-characterized controls, will help to calibrate doses to compare preclinical and clinical data in the field.

Construction of new cloning, lacZ reporter and scarless-markerless suicide vectors for genetic studies in Aggregatibacter actinomycetemcomitans

PubMed Central

Juárez-Rodríguez, María Dolores; Torres-Escobar, Ascención; Demuth, Donald R.

2013-01-01

To elucidate the putative function of a gene, effective tools are required for genetic characterization that facilitate its inactivation, deletion or modification on the bacterial chromosome. In the present study, the nucleotide sequence of the Escherichia coli/Aggregatibacter actinomycetemcomitans shuttle vector pYGK was determined, allowing us to redesign and construct a new shuttle cloning vector, pJT4, and promoterless lacZ transcriptional/translational fusion plasmids, pJT3 and pJT5. Plasmids pJT4 and pJT5 contain the origin of replication necessary to maintain shuttle vector replication. In addition, a new suicide vector, pJT1, was constructed for the generation of scarless and markerless deletion mutations of genes in the oral pathogen A. actinomycetemcomitans. Plasmid pJT1 is a pUC-based suicide vector that is counter-selectable for sucrose sensitivity. This vector does not leave antibiotic markers or scars on the chromosome after gene deletion and thus provides the option to combine several mutations in the same genetic background. The effectiveness of pJT1 was demonstrated by the construction of A. actinomycetemcomitans isogenic qseB single deletion (ΔqseB) mutant and lsrRK double deletion mutants (ΔlsrRK). These new vectors may offer alternatives for genetic studies in A. actinomycetemcomitans and other members of the HACEK (Haemophilus spp., A. actinomycetemcomitans, Cardiobacterium hominis, Eikenella corrodens, and Kingella kingae) group of Gram-negative bacteria. PMID:23353051

Construction and applications of exon-trapping gene-targeting vectors with a novel strategy for negative selection.

PubMed

Saito, Shinta; Ura, Kiyoe; Kodama, Miho; Adachi, Noritaka

2015-06-30

Targeted gene modification by homologous recombination provides a powerful tool for studying gene function in cells and animals. In higher eukaryotes, non-homologous integration of targeting vectors occurs several orders of magnitude more frequently than does targeted integration, making the gene-targeting technology highly inefficient. For this reason, negative-selection strategies have been employed to reduce the number of drug-resistant clones associated with non-homologous vector integration, particularly when artificial nucleases to introduce a DNA break at the target site are unavailable or undesirable. As such, an exon-trap strategy using a promoterless drug-resistance marker gene provides an effective way to counterselect non-homologous integrants. However, constructing exon-trapping targeting vectors has been a time-consuming and complicated process. By virtue of highly efficient att-mediated recombination, we successfully developed a simple and rapid method to construct plasmid-based vectors that allow for exon-trapping gene targeting. These exon-trap vectors were useful in obtaining correctly targeted clones in mouse embryonic stem cells and human HT1080 cells. Most importantly, with the use of a conditionally cytotoxic gene, we further developed a novel strategy for negative selection, thereby enhancing the efficiency of counterselection for non-homologous integration of exon-trap vectors. Our methods will greatly facilitate exon-trapping gene-targeting technologies in mammalian cells, particularly when combined with the novel negative selection strategy.

Double-stranded RNA innate immune response activation from long-term adeno-associated virus vector transduction.

PubMed

Shao, Wenwei; Earley, Lauriel F; Chai, Zheng; Chen, Xiaojing; Sun, Junjiang; He, Ting; Deng, Meng; Hirsch, Matthew L; Ting, Jenny; Samulski, R Jude; Li, Chengwen

2018-06-21

Data from clinical trials for hemophilia B using adeno-associated virus (AAV) vectors have demonstrated decreased transgenic coagulation factor IX (hFIX) expression 6-10 weeks after administration of a high vector dose. While it is likely that capsid-specific cytotoxic T lymphocytes eliminate vector-transduced hepatocytes, thereby resulting in decreased hFIX, this observation is not intuitively consistent with restored hFIX levels following prednisone application. Although the innate immune response is immediately activated following AAV vector infection via TLR pathways, no studies exist regarding the role of the innate immune response at later time points after AAV vector transduction. Herein, activation of the innate immune response in cell lines, primary human hepatocytes, and hepatocytes in a human chimeric mouse model was observed at later time points following AAV vector transduction. Mechanistic analysis demonstrated that the double-stranded RNA (dsRNA) sensor MDA5 was necessary for innate immune response activation and that transient knockdown of MDA5, or MAVS, decreased IFN-β expression while increasing transgene production in AAV-transduced cells. These results both highlight the role of the dsRNA-triggered innate immune response in therapeutic transgene expression at later time points following AAV transduction and facilitate the execution of effective strategies to block the dsRNA innate immune response in future clinical trials.

Vector production in an academic environment: a tool to assess production costs.

PubMed

Boeke, Aaron; Doumas, Patrick; Reeves, Lilith; McClurg, Kyle; Bischof, Daniela; Sego, Lina; Auberry, Alisha; Tatikonda, Mohan; Cornetta, Kenneth

2013-02-01

Generating gene and cell therapy products under good manufacturing practices is a complex process. When determining the cost of these products, researchers must consider the large number of supplies used for manufacturing and the personnel and facility costs to generate vector and maintain a cleanroom facility. To facilitate cost estimates, the Indiana University Vector Production Facility teamed with the Indiana University Kelley School of Business to develop a costing tool that, in turn, provides pricing. The tool is designed in Microsoft Excel and is customizable to meet the needs of other core facilities. It is available from the National Gene Vector Biorepository. The tool allows cost determinations using three different costing methods and was developed in an effort to meet the A21 circular requirements for U.S. core facilities performing work for federally funded projects. The costing tool analysis reveals that the cost of vector production does not have a linear relationship with batch size. For example, increasing the production from 9 to18 liters of a retroviral vector product increases total costs a modest 1.2-fold rather than doubling in total cost. The analysis discussed in this article will help core facilities and investigators plan a cost-effective strategy for gene and cell therapy production.

Multiplexing clonality: combining RGB marking and genetic barcoding

PubMed Central

Cornils, Kerstin; Thielecke, Lars; Hüser, Svenja; Forgber, Michael; Thomaschewski, Michael; Kleist, Nadja; Hussein, Kais; Riecken, Kristoffer; Volz, Tassilo; Gerdes, Sebastian; Glauche, Ingmar; Dahl, Andreas; Dandri, Maura; Roeder, Ingo; Fehse, Boris

2014-01-01

RGB marking and DNA barcoding are two cutting-edge technologies in the field of clonal cell marking. To combine the virtues of both approaches, we equipped LeGO vectors encoding red, green or blue fluorescent proteins with complex DNA barcodes carrying color-specific signatures. For these vectors, we generated highly complex plasmid libraries that were used for the production of barcoded lentiviral vector particles. In proof-of-principle experiments, we used barcoded vectors for RGB marking of cell lines and primary murine hepatocytes. We applied single-cell polymerase chain reaction to decipher barcode signatures of individual RGB-marked cells expressing defined color hues. This enabled us to prove clonal identity of cells with one and the same RGB color. Also, we made use of barcoded vectors to investigate clonal development of leukemia induced by ectopic oncogene expression in murine hematopoietic cells. In conclusion, by combining RGB marking and DNA barcoding, we have established a novel technique for the unambiguous genetic marking of individual cells in the context of normal regeneration as well as malignant outgrowth. Moreover, the introduction of color-specific signatures in barcodes will facilitate studies on the impact of different variables (e.g. vector type, transgenes, culture conditions) in the context of competitive repopulation studies. PMID:24476916

Vector Production in an Academic Environment: A Tool to Assess Production Costs

PubMed Central

Boeke, Aaron; Doumas, Patrick; Reeves, Lilith; McClurg, Kyle; Bischof, Daniela; Sego, Lina; Auberry, Alisha; Tatikonda, Mohan

2013-01-01

Abstract Generating gene and cell therapy products under good manufacturing practices is a complex process. When determining the cost of these products, researchers must consider the large number of supplies used for manufacturing and the personnel and facility costs to generate vector and maintain a cleanroom facility. To facilitate cost estimates, the Indiana University Vector Production Facility teamed with the Indiana University Kelley School of Business to develop a costing tool that, in turn, provides pricing. The tool is designed in Microsoft Excel and is customizable to meet the needs of other core facilities. It is available from the National Gene Vector Biorepository. The tool allows cost determinations using three different costing methods and was developed in an effort to meet the A21 circular requirements for U.S. core facilities performing work for federally funded projects. The costing tool analysis reveals that the cost of vector production does not have a linear relationship with batch size. For example, increasing the production from 9 to18 liters of a retroviral vector product increases total costs a modest 1.2-fold rather than doubling in total cost. The analysis discussed in this article will help core facilities and investigators plan a cost-effective strategy for gene and cell therapy production. PMID:23360377

On sufficient statistics of least-squares superposition of vector sets.

PubMed

Konagurthu, Arun S; Kasarapu, Parthan; Allison, Lloyd; Collier, James H; Lesk, Arthur M

2015-06-01

The problem of superposition of two corresponding vector sets by minimizing their sum-of-squares error under orthogonal transformation is a fundamental task in many areas of science, notably structural molecular biology. This problem can be solved exactly using an algorithm whose time complexity grows linearly with the number of correspondences. This efficient solution has facilitated the widespread use of the superposition task, particularly in studies involving macromolecular structures. This article formally derives a set of sufficient statistics for the least-squares superposition problem. These statistics are additive. This permits a highly efficient (constant time) computation of superpositions (and sufficient statistics) of vector sets that are composed from its constituent vector sets under addition or deletion operation, where the sufficient statistics of the constituent sets are already known (that is, the constituent vector sets have been previously superposed). This results in a drastic improvement in the run time of the methods that commonly superpose vector sets under addition or deletion operations, where previously these operations were carried out ab initio (ignoring the sufficient statistics). We experimentally demonstrate the improvement our work offers in the context of protein structural alignment programs that assemble a reliable structural alignment from well-fitting (substructural) fragment pairs. A C++ library for this task is available online under an open-source license.

Plasmids containing the gene for DNA polymerase I from Streptococcus pneumoniae

DOEpatents

Lacks, S.A.; Martinez, S.; Lopez, P.; Espinosa, M.

1991-03-26

A method is disclosed for cloning the gene which encodes a DNA polymerase-exonuclease of Streptococcus pneumoniae. Plasmid pSM22, the vector containing the pneumocccal polA gene, facilitates the expression of 50-fold greater amounts of the PolI enzyme. 1 figure.

Surface coating of siRNA-peptidomimetic nano-self-assemblies with anionic lipid bilayers: enhanced gene silencing and reduced adverse effects in vitro

NASA Astrophysics Data System (ADS)

Zeng, Xianghui; de Groot, Anne Marit; Sijts, Alice J. A. M.; Broere, Femke; Oude Blenke, Erik; Colombo, Stefano; van Eden, Willem; Franzyk, Henrik; Nielsen, Hanne Mørck; Foged, Camilla

2015-11-01

Cationic vectors have demonstrated the potential to facilitate intracellular delivery of therapeutic oligonucleotides. However, enhanced transfection efficiency is usually associated with adverse effects, which also proves to be a challenge for vectors based on cationic peptides. In this study a series of proteolytically stable palmitoylated α-peptide/β-peptoid peptidomimetics with a systematically varied number of repeating lysine and homoarginine residues was shown to self-assemble with small interfering RNA (siRNA). The resulting well-defined nanocomplexes were coated with anionic lipids giving rise to net anionic liposomes. These complexes and the corresponding liposomes were optimized towards efficient gene silencing and low adverse effects. The optimal anionic liposomes mediated a high silencing effect, which was comparable to that of the control (cationic Lipofectamine 2000), and did not display any noticeable cytotoxicity and immunogenicity in vitro. In contrast, the corresponding nanocomplexes mediated a reduced silencing effect with a more narrow safety window. The surface coating with anionic lipid bilayers led to partial decomplexation of the siRNA-peptidomimetic nanocomplex core of the liposomes, which facilitated siRNA release. Additionally, the optimal anionic liposomes showed efficient intracellular uptake and endosomal escape. Therefore, these findings suggest that a more efficacious and safe formulation can be achieved by surface coating of the siRNA-peptidomimetic nano-self-assemblies with anionic lipid bilayers.Cationic vectors have demonstrated the potential to facilitate intracellular delivery of therapeutic oligonucleotides. However, enhanced transfection efficiency is usually associated with adverse effects, which also proves to be a challenge for vectors based on cationic peptides. In this study a series of proteolytically stable palmitoylated α-peptide/β-peptoid peptidomimetics with a systematically varied number of repeating lysine and homoarginine residues was shown to self-assemble with small interfering RNA (siRNA). The resulting well-defined nanocomplexes were coated with anionic lipids giving rise to net anionic liposomes. These complexes and the corresponding liposomes were optimized towards efficient gene silencing and low adverse effects. The optimal anionic liposomes mediated a high silencing effect, which was comparable to that of the control (cationic Lipofectamine 2000), and did not display any noticeable cytotoxicity and immunogenicity in vitro. In contrast, the corresponding nanocomplexes mediated a reduced silencing effect with a more narrow safety window. The surface coating with anionic lipid bilayers led to partial decomplexation of the siRNA-peptidomimetic nanocomplex core of the liposomes, which facilitated siRNA release. Additionally, the optimal anionic liposomes showed efficient intracellular uptake and endosomal escape. Therefore, these findings suggest that a more efficacious and safe formulation can be achieved by surface coating of the siRNA-peptidomimetic nano-self-assemblies with anionic lipid bilayers. Electronic supplementary information (ESI) available: Non-fusogenic liposomes; cytotoxicity of naked siRNA and the empty vector; immunogenicity; low-magnification images; DOPE/DPPC liposomes. See DOI: 10.1039/c5nr04807a

Cre-lox Univector acceptor vectors for functional screening in protoplasts: analysis of Arabidopsis donor cDNAs encoding ABSCISIC ACID INSENSITIVE1-Like protein phosphatases

PubMed Central

Jia, Fan; Gampala, Srinivas S.L.; Mittal, Amandeep; Luo, Qingjun; Rock, Christopher D.

2009-01-01

The 14,200 available full length Arabidopsis thaliana cDNAs in the Universal Plasmid System (UPS) donor vector pUNI51 should be applied broadly and efficiently to leverage a “functional map-space” of homologous plant genes. We have engineered Cre-lox UPS host acceptor vectors (pCR701- 705) with N-terminal epitope tags in frame with the loxH site and downstream from the maize Ubiquitin promoter for use in transient protoplast expression assays and particle bombardment transformation of monocots. As an example of the utility of these vectors, we recombined them with several Arabidopsis cDNAs encoding Ser/Thr protein phosphatase type 2C (PP2Cs) known from genetic studies or predicted by hierarchical clustering meta-analysis to be involved in ABA and stress responses. Our functional results in Zea mays mesophyll protoplasts on ABA-inducible expression effects on the Late Embryogenesis Abundant promoter ProEm:GUS reporter were consistent with predictions and resulted in identification of novel activities of some PP2Cs. Deployment of these vectors can facilitate functional genomics and proteomics and identification of novel gene activities. PMID:19499346

New Drug Formulary Will Help Expedite Use of Agents in Clinical Trials

Cancer.gov

NCI launched the “NCI Formulary” that will enable investigators at NCI-designated Cancer Centers to have quicker access to approved and investigational agents for use in preclinical studies and cancer clinical trials.

Use of mobile devices to answer online surveys: implications for research.

PubMed

Cunningham, John A; Neighbors, Clayton; Bertholet, Nicolas; Hendershot, Christian S

2013-07-08

There is a growing use of mobile devices to access the Internet. We examined whether participants who used a mobile device to access a brief online survey were quicker to respond to the survey but also, less likely to complete it than participants using a traditional web browser. Using data from a recently completed online intervention trial, we found that participants using mobile devices were quicker to access the survey but less likely to complete it compared to participants using a traditional web browser. More concerning, mobile device users were also less likely to respond to a request to complete a six week follow-up survey compared to those using traditional web browsers. With roughly a third of participants using mobile devices to answer an online survey in this study, the impact of mobile device usage on survey completion rates is a concern. ClinicalTrials.gov: NCT01521078.

Large spot transpupillary thermotherapy: a quicker laser for treatment of high risk prethreshold retinopathy of prematurity - a randomized study.

PubMed

Shah, Parag K; Narendran, V; Kalpana, N

2011-01-01

To compare structural and functional outcome and time efficiency between standard spot sized conventional pulsed mode diode laser and continuous mode large spot transpupillary thermotherapy (LS TTT) for treatment of high risk prethreshold retinopathy of prematurity (ROP). Ten eyes of five preterm babies having bilateral symmetrical high risk prethreshold ROP were included in this study. One eye of each baby was randomized to get either standard spot sized conventional pulsed mode diode laser or continuous mode LS TTT. There was no significant difference between structural or functional outcome in either group. The mean time taken for conventional diode laser was 20.07 minutes, while that for LS TTT was 12.3 minutes. LS TTT was 40% more time efficient than the conventional laser. It may be better suited for the very small fragile premature infants as it is quicker than the conventional laser.

Plasmids containing the gene for DNA polymerase I from Streptococcus pneumoniae

DOEpatents

Lacks, S.A.; Martinez, S.; Lopez, P.; Espinosa, M.

1987-08-28

A method is disclosed for cloning the gene which encodes a DNA polymerase-exonuclease of /und Streptococcus/ /und pneumoniae/. Plasmid pSM22, the vector containing the pneumococcal polA gene, facilitates the expression of 50-fold greater amounts of the PolI enzyme. 1 fig., 1 tab.

The C-terminus of the Polerovirus P5 readthrough domain limits virus infection to the phloem

USDA-ARS?s Scientific Manuscript database

Unlike most plant viruses, poleroviruses are restricted to vascular phloem tissues, from which they are transmitted by their aphid vectors. Phloem limitation has been attributed to the absence of virus proteins facilitating movement or counteracting plant defense. The polerovirus capsid is composed ...

78 FR 65556 - Establishment of Class E Airspace; Cut Bank, MT

Federal Register 2010, 2011, 2012, 2013, 2014

2013-11-01

...-0532; Airspace Docket No. 13-ANM-21] Establishment of Class E Airspace; Cut Bank, MT AGENCY: Federal... at the Cut Bank VHF Omni-Directional Radio Range Tactical Air Navigational Aid (VORTAC) navigation aid, Cut Bank, MT, to facilitate vectoring of Instrument Flight Rules (IFR) aircraft under control of...

Manipulating the Gradient

ERIC Educational Resources Information Center

Gaze, Eric C.

2005-01-01

We introduce a cooperative learning, group lab for a Calculus III course to facilitate comprehension of the gradient vector and directional derivative concepts. The lab is a hands-on experience allowing students to manipulate a tangent plane and empirically measure the effect of partial derivatives on the direction of optimal ascent. (Contains 7…

78 FR 78298 - Proposed Establishment of Class E Airspace; Phoenix, AZ

Federal Register 2010, 2011, 2012, 2013, 2014

2013-12-26

...-0956; Airspace Docket No. 13-AWP-17] Proposed Establishment of Class E Airspace; Phoenix, AZ AGENCY... rulemaking (NPRM). SUMMARY: This action proposes to establish Class E airspace at the Phoenix VHF Omni-Directional Radio Range Tactical Air Navigation Aid (VORTAC), Phoenix, AZ, to facilitate vectoring of...

A hitchhiker's guide to the Maritimes: anthropogenic transport facilitates long-distance dispersal of an invasive marine crab to Newfoundland

EPA Science Inventory

We analyzed mitochondrial and microsatellite genetic data from European and Northwest Atlantic populations sampled during 1999-2002 to determine probable source locations and vectors for the Placentia Bay introduction discovered in 2007. We also analyzed Placentia Bay demographi...

78 FR 78299 - Proposed Establishment of Class E Airspace; Truth or Consequences, NM

Federal Register 2010, 2011, 2012, 2013, 2014

2013-12-26

...-0995; Airspace Docket No. 13-ASW-30] Proposed Establishment of Class E Airspace; Truth or Consequences... Truth or Consequences VHF Omni-Directional Radio Range Tactical Air Navigation Aid (VORTAC), Truth or Consequences, NM, to facilitate vectoring of Instrument Flight Rules (IFR) aircraft under control of...

78 FR 65555 - Establishment of Class E Airspace; Salmon, ID

Federal Register 2010, 2011, 2012, 2013, 2014

2013-11-01

...-0531; Airspace Docket No. 13-ANM-20] Establishment of Class E Airspace; Salmon, ID AGENCY: Federal... at the Salmon VHF Omni-Directional Radio Range/Distance Measuring Equipment (VOR/DME) navigation aid, Salmon, ID, to facilitate vectoring of Instrument Flight Rules (IFR) aircraft under control of Salt Lake...

78 FR 45478 - Proposed Establishment of Class E Airspace; Salmon, ID

Federal Register 2010, 2011, 2012, 2013, 2014

2013-07-29

...-0531; Airspace Docket No. 13-ANM-20] Proposed Establishment of Class E Airspace; Salmon, ID AGENCY... action proposes to establish Class E airspace at the Salmon VHF Omni-Directional Radio Range/Distance Measuring Equipment (VOR/DME) navigation aid, Salmon, ID, to facilitate vectoring of Instrument Flight Rules...

Simplified production and concentration of HIV-1-based lentiviral vectors using HYPERFlask vessels and anion exchange membrane chromatography

PubMed Central

Kutner, Robert H; Puthli, Sharon; Marino, Michael P; Reiser, Jakob

2009-01-01

Background During the past twelve years, lentiviral (LV) vectors have emerged as valuable tools for transgene delivery because of their ability to transduce nondividing cells and their capacity to sustain long-term transgene expression in target cells in vitro and in vivo. However, despite significant progress, the production and concentration of high-titer, high-quality LV vector stocks is still cumbersome and costly. Methods Here we present a simplified protocol for LV vector production on a laboratory scale using HYPERFlask vessels. HYPERFlask vessels are high-yield, high-performance flasks that utilize a multilayered gas permeable growth surface for efficient gas exchange, allowing convenient production of high-titer LV vectors. For subsequent concentration of LV vector stocks produced in this way, we describe a facile protocol involving Mustang Q anion exchange membrane chromatography. Results Our results show that unconcentrated LV vector stocks with titers in excess of 108 transduction units (TU) per ml were obtained using HYPERFlasks and that these titers were higher than those produced in parallel using regular 150-cm2 tissue culture dishes. We also show that up to 500 ml of an unconcentrated LV vector stock prepared using a HYPERFlask vessel could be concentrated using a single Mustang Q Acrodisc with a membrane volume of 0.18 ml. Up to 5.3 × 1010 TU were recovered from a single HYPERFlask vessel. Conclusion The protocol described here is easy to implement and should facilitate high-titer LV vector production for preclinical studies in animal models without the need for multiple tissue culture dishes and ultracentrifugation-based concentration protocols. PMID:19220915

Modification and identification of a vector for making a large phage antibody library.

PubMed

Zhang, Guo-min; Chen, Yü-ping; Guan, Yuan-zhi; Wang, Yan; An, Yun-qing

2007-11-20

The large phage antibody library is used to obtain high-affinity human antibody, and the Loxp/cre site-specific recombination system is a potential method for constructing a large phage antibody library. In the present study, a phage antibody library vector pDF was reconstructed to construct diabody more quickly and conveniently without injury to homologous recombination and the expression function of the vector and thus to integrate construction of the large phage antibody library with the preparation of diabodies. scFv was obtained by overlap polymerase chain reaction (PCR) amplification with the newly designed VL and VH extension primers. loxp511 was flanked by VL and VH and the endonuclease ACC III encoding sequences were introduced on both sides of loxp511. scFv was cloned into the vector pDF to obtain the vector pDscFv. The vector expression function was identified and the feasibility of diabody preparation was evaluated. A large phage antibody library was constructed in pDscFv. Several antigens were used to screen the antibody library and the quality of the antibody library was evaluated. The phage antibody library expression vector pDscFv was successfully constructed and confirmed to express functional scFv. The large phage antibody library constructed using this vector was of high diversity. Screening of the library on 6 antigens confirmed the generation of specific antibodies to these antigens. Two antibodies were subjected to enzymatic digestion and were prepared into diabody with functional expression. The reconstructed vector pDscFv retains its recombination capability and expression function and can be used to construct large phage antibody libraries. It can be used as a convenient and quick method for preparing diabodies after simple enzymatic digestion, which facilitates clinical trials and application of antibody therapy.

3-minute diagnosis: Researchers develop new method to recognize pathogens

DOE Office of Scientific and Technical Information (OSTI.GOV)

Beer, Reg

Imagine knowing precisely why you feel sick ... before the doctor's exam is over. Lawrence Livermore researcher Reg Beer and his engineering colleagues have developed a new method to recognize disease-causing pathogens quicker than ever before.

3-minute diagnosis: Researchers develop new method to recognize pathogens

ScienceCinema

Beer, Reg

2018-01-16

Imagine knowing precisely why you feel sick ... before the doctor's exam is over. Lawrence Livermore researcher Reg Beer and his engineering colleagues have developed a new method to recognize disease-causing pathogens quicker than ever before.

Thermodynamic Analysis of a Coupled Chemical Reaction.

ERIC Educational Resources Information Center

Trimm, Harold; And Others

1979-01-01

Describes a typical relaxation kinetic experiment using a sudden increase in the temperature of the system. Time involved is described as minimal and the approach as quicker, more accurate, sensitive, and producing simultaneous determination of several thermodynamic parameters. (Author/SA)

Hysteroscopic Sterilization

MedlinePlus

... sterilization? Sterilization is a permanent form of birth control. What is tubal sterilization? Sterilization procedures for women are ... is quicker than from other types of sterilization. What are the risks of ... on for birth control. • There is a risk of injury to the ...

Turbulent Recirculating Flows in Isothermal Combustor Geometries

NASA Technical Reports Server (NTRS)

Lilley, D.; Rhode, D.

1985-01-01

Computer program developed that provides mathematical solution to design and construction of combustion chambers for jet engines. Improved results in areas of combustor flow fields accomplished by this computerprogram solution, cheaper and quicker than experiments involving real systems for models.

Tyrosine-phosphorylation of AAV2 vectors and its consequences on viral intracellular trafficking and transgene expression

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zhong Li; Powell Gene Therapy Center, University of Florida College of Medicine, Gainesville, FL; Genetics Institute, University of Florida College of Medicine, Gainesville, FL

2008-11-25

We have documented that epidermal growth factor receptor protein tyrosine kinase (EGFR-PTK) signaling negatively affects intracellular trafficking and transduction efficiency of recombinant adeno-associated virus 2 (AAV2) vectors. Specifically, inhibition of EGFR-PTK signaling leads to decreased ubiquitination of AAV2 capsid proteins, which in turn, facilitates viral nuclear transport by limiting proteasome-mediated degradation of AAV2 vectors. In the present studies, we observed that AAV capsids can indeed be phosphorylated at tyrosine residues by EGFR-PTK in in vitro phosphorylation assays and that phosphorylated AAV capsids retain their structural integrity. However, although phosphorylated AAV vectors enter cells as efficiently as their unphosphorylated counterparts, theirmore » transduction efficiency is significantly reduced. This reduction is not due to impaired viral second-strand DNA synthesis since transduction efficiency of both single-stranded AAV (ssAAV) and self-complementary AAV (scAAV) vectors is decreased by {approx} 68% and {approx} 74%, respectively. We also observed that intracellular trafficking of tyrosine-phosphorylated AAV vectors from cytoplasm to nucleus is significantly decreased, which results from ubiquitination of AAV capsids followed by proteasome-mediated degradation, although downstream consequences of capsid ubiquitination may also be affected by tyrosine-phosphorylation. These studies provide new insights into the role of tyrosine-phosphorylation of AAV capsids in various steps in the virus life cycle, which has implications in the optimal use of recombinant AAV vectors in human gene therapy.« less

The Extinction of Dengue through Natural Vulnerability of Its Vectors

PubMed Central

Williams, Craig R.; Bader, Christie A.; Kearney, Michael R.; Ritchie, Scott A.; Russell, Richard C.

2010-01-01

Background Dengue is the world's most important mosquito-borne viral illness. Successful future management of this disease requires an understanding of the population dynamics of the vector, especially in the context of changing climates. Our capacity to predict future dynamics is reflected in our ability to explain the significant historical changes in the distribution and abundance of the disease and its vector. Methodology/Principal Findings Here we combine daily weather records with simulation modelling techniques to explain vector (Aedes aegypti (L.)) persistence within its current and historic ranges in Australia. We show that, in regions where dengue presently occurs in Australia (the Wet Tropics region of Far North Queensland), conditions are persistently suitable for year-round adult Ae. aegypti activity and oviposition. In the historic range, however, the vector is vulnerable to periodic extinction due to the combined influence of adult activity constraints and stochastic loss of suitable oviposition sites. Conclusions/Significance These results, together with changes in water-storage behaviour by humans, can explain the observed historical range contraction of the disease vector. For these reasons, future eradication of dengue in wet tropical regions will be extremely difficult through classical mosquito control methods alone. However, control of Ae. aegypti in sub-tropical and temperate regions will be greatly facilitated by government policy regulating domestic water-storage. Exploitation of the natural vulnerabilities of dengue vectors (e.g., habitat specificity, climatic limitations) should be integrated with the emerging novel transgenic and symbiotic bacterial control techniques to develop future control and elimination strategies. PMID:21200424

Vector-based genetically modified vaccines: Exploiting Jenner's legacy.

PubMed

Ramezanpour, Bahar; Haan, Ingrid; Osterhaus, Ab; Claassen, Eric

2016-12-07

The global vaccine market is diverse while facing a plethora of novel developments. Genetic modification (GM) techniques facilitate the design of 'smarter' vaccines. For many of the major infectious diseases of humans, like AIDS and malaria, but also for most human neoplastic disorders, still no vaccines are available. It may be speculated that novel GM technologies will significantly contribute to their development. While a promising number of studies is conducted on GM vaccines and GM vaccine technologies, the contribution of GM technology to newly introduced vaccines on the market is disappointingly limited. In this study, the field of vector-based GM vaccines is explored. Data on currently available, actually applied, and newly developed vectors is retrieved from various sources, synthesised and analysed, in order to provide an overview on the use of vector-based technology in the field of GM vaccine development. While still there are only two vector-based vaccines on the human vaccine market, there is ample activity in the fields of patenting, preclinical research, and different stages of clinical research. Results of this study revealed that vector-based vaccines comprise a significant part of all GM vaccines in the pipeline. This study further highlights that poxviruses and adenoviruses are among the most prominent vectors in GM vaccine development. After the approval of the first vectored human vaccine, based on a flavivirus vector, vaccine vector technology, especially based on poxviruses and adenoviruses, holds great promise for future vaccine development. It may lead to cheaper methods for the production of safe vaccines against diseases for which no or less perfect vaccines exist today, thus catering for an unmet medical need. After the introduction of Jenner's vaccinia virus as the first vaccine more than two centuries ago, which eventually led to the recent eradication of smallpox, this and other viruses may now be the basis for constructing vectors that may help us control other major scourges of mankind. Copyright © 2016 The Author(s). Published by Elsevier Ltd.. All rights reserved.

Mouse mammary tumor virus-based vector transduces non-dividing cells, enters the nucleus via a TNPO3-independent pathway and integrates in a less biased fashion than other retroviruses.

PubMed

Konstantoulas, Constantine James; Indik, Stanislav

2014-04-30

Mouse mammary tumor virus (MMTV) is a complex, milk-born betaretrovirus, which preferentially infects dendritic cells (DC) in the gastrointestinal tract and then spreads to T and B lymphocytes and finally to the mammary gland. It is not clear how the prototypic betaretrovirus infects mucosal DCs and naïve lymphocytes as these cells are considered to be non-proliferative. Studies of MMTV biology have been hampered by the difficulty of obtaining sufficient virus/vector titers after transfection of a molecular clone in cultured cells. To surmount this barrier we developed a novel MMTV-based vector system with a split genome design containing potent posttranscriptional regulatory functions. Using this system, vector particles were produced to markedly greater titers (>1000-fold) than those obtained previously. The titers (>106 transduction units /ml) were comparable to those achieved with lentiviral or gammaretroviral vectors. Importantly, the vector transduced the enhanced green fluorescence protein gene into the chromosomes of non-dividing cells, such as cells arrested at the G2/M phase of the cell cycle and unstimulated hematopoietic progenitor cells, at an efficiency similar to that obtained with the HIV-1-based vector. In contrast to HIV-1, MMTV transductions were not affected by knocking down the expression of a factor involved in nuclear import of the HIV-1 pre-integration complexes, TNPO3. In contrast to HIV-1, the MMTV-based vector did not preferentially integrate in transcription units. Additionally, no preference for integration near transcription start sites, the regions preferentially targeted by gammaretroviral vectors, was observed. The vector derived from MMTV exhibits a random integration pattern. Overall, the betaretroviral vector system should facilitate molecular virology studies of the prototypic betaretrovirus as well as studies attempting to elucidate fundamental cellular processes such as nuclear import pathways. Random integration in cycling and non-cycling cells may be applicable in unbiased gene delivery.

Vector Observation-Aided/Attitude-Rate Estimation Using Global Positioning System Signals

NASA Technical Reports Server (NTRS)

Oshman, Yaakov; Markley, F. Landis

1997-01-01

A sequential filtering algorithm is presented for attitude and attitude-rate estimation from Global Positioning System (GPS) differential carrier phase measurements. A third-order, minimal-parameter method for solving the attitude matrix kinematic equation is used to parameterize the filter's state, which renders the resulting estimator computationally efficient. Borrowing from tracking theory concepts, the angular acceleration is modeled as an exponentially autocorrelated stochastic process, thus avoiding the use of the uncertain spacecraft dynamic model. The new formulation facilitates the use of aiding vector observations in a unified filtering algorithm, which can enhance the method's robustness and accuracy. Numerical examples are used to demonstrate the performance of the method.

Data catalog for JPL Physical Oceanography Distributed Active Archive Center (PO.DAAC)

NASA Technical Reports Server (NTRS)

Digby, Susan

1995-01-01

The Physical Oceanography Distributed Active Archive Center (PO.DAAC) archive at the Jet Propulsion Laboratory contains satellite data sets and ancillary in-situ data for the ocean sciences and global-change research to facilitate multidisciplinary use of satellite ocean data. Geophysical parameters available from the archive include sea-surface height, surface-wind vector, surface-wind speed, surface-wind stress vector, sea-surface temperature, atmospheric liquid water, integrated water vapor, phytoplankton pigment concentration, heat flux, and in-situ data. PO.DAAC is an element of the Earth Observing System Data and Information System and is the United States distribution site for TOPEX/POSEIDON data and metadata.

Phagemid vectors for phage display: properties, characteristics and construction.

PubMed

Qi, Huan; Lu, Haiqin; Qiu, Hua-Ji; Petrenko, Valery; Liu, Aihua

2012-03-30

Phagemids are filamentous-phage-derived vectors containing the replication origin of a plasmid. Phagemids usually encode no or only one kind of coat proteins. Other structural and functional proteins necessary to accomplish the life cycle of phagemid are provided by the helper phage. In addition, other elements such as molecular tags and selective markers are introduced into the phagemids to facilitate the subsequent operations, such as gene manipulation and protein purification. This review summarizes the elements of the phagemids and their corresponding functions. Finally, the possible trends and future direction to improve the characteristics of the phagemids are highlighted. Copyright © 2012 Elsevier Ltd. All rights reserved.

Spatial inventory integrating raster databases and point sample data. [Geographic Information System for timber inventory

NASA Technical Reports Server (NTRS)

Strahler, A. H.; Woodcock, C. E.; Logan, T. L.

1983-01-01

A timber inventory of the Eldorado National Forest, located in east-central California, provides an example of the use of a Geographic Information System (GIS) to stratify large areas of land for sampling and the collection of statistical data. The raster-based GIS format of the VICAR/IBIS software system allows simple and rapid tabulation of areas, and facilitates the selection of random locations for ground sampling. Algorithms that simplify the complex spatial pattern of raster-based information, and convert raster format data to strings of coordinate vectors, provide a link to conventional vector-based geographic information systems.

Assessment of ICount software, a precise and fast egg counting tool for the mosquito vector Aedes aegypti.

PubMed

Gaburro, Julie; Duchemin, Jean-Bernard; Paradkar, Prasad N; Nahavandi, Saeid; Bhatti, Asim

2016-11-18

Widespread in the tropics, the mosquito Aedes aegypti is an important vector of many viruses, posing a significant threat to human health. Vector monitoring often requires fecundity estimation by counting eggs laid by female mosquitoes. Traditionally, manual data analyses have been used but this requires a lot of effort and is the methods are prone to errors. An easy tool to assess the number of eggs laid would facilitate experimentation and vector control operations. This study introduces a built-in software called ICount allowing automatic egg counting of the mosquito vector, Aedes aegypti. ICount egg estimation compared to manual counting is statistically equivalent, making the software effective for automatic and semi-automatic data analysis. This technique also allows rapid analysis compared to manual methods. Finally, the software has been used to assess p-cresol oviposition choices under laboratory conditions in order to test the system with different egg densities. ICount is a powerful tool for fast and precise egg count analysis, freeing experimenters from manual data processing. Software access is free and its user-friendly interface allows easy use by non-experts. Its efficiency has been tested in our laboratory with oviposition dual choices of Aedes aegypti females. The next step will be the development of a mobile application, based on the ICount platform, for vector monitoring surveys in the field.

Plasmid Vectors for Xylella fastidiosa Utilizing a Toxin-Antitoxin System for Stability in the Absence of Antibiotic Selection.

PubMed

Burbank, Lindsey P; Stenger, Drake C

2016-08-01

The phytopathogen Xylella fastidiosa causes disease in a variety of important crop and landscape plants. Functional genetic studies have led to a broader understanding of virulence mechanisms used by this pathogen in the grapevine host. Plasmid shuttle vectors are important tools in studies of bacterial genetics but there are only a limited number of plasmid vectors available that replicate in X. fastidiosa, and even fewer that are retained without antibiotic selection. Two plasmids are described here that show stable replication in X. fastidiosa and are effective for gene complementation both in vitro and in planta. Plasmid maintenance is facilitated by incorporation of the PemI/PemK plasmid addiction system, consisting of PemK, an endoribonuclease toxin, and its cognate antitoxin, PemI. Vector pXf20pemIK utilizes a native X. fastidiosa replication origin as well as a high-copy-number pUC origin for propagation in Escherichia coli cloning strains. Broad-host-range vector pBBR5pemIK is a medium- to low-copy-number plasmid based on the pBBR1 backbone. Both plasmids are maintained for extended periods of time in the absence of antibiotic selection, as well as up to 14 weeks in grapevine, without affecting bacterial fitness. These plasmids present an alternative to traditional complementation and expression vectors which rely on antibiotic selection for plasmid retention.

Development of a pan-rickettsial molecular diagnostic test based on recombinase polymerase amplification assay.

PubMed

Kissenkötter, Jonas; Hansen, Sören; Böhlken-Fascher, Susanne; Ademowo, Olusegun George; Oyinloye, Oladapo Elijah; Bakarey, Adeleye Solomon; Dobler, Gerhard; Tappe, Dennis; Patel, Pranav; Czerny, Claus-Peter; Abd El Wahed, Ahmed

2018-03-01

Rickettsioses are zoonotic vector-transmitted bacterial infections leading to flu-like symptoms and can progress to severe illness in humans. The gold standard for diagnosis of rickettsial infections is the indirect immunofluorescence assay, a serological method which is not suitable for pathogen identification during the acute phase of the disease. Therefore, several real-time PCR assays were developed. These assays are very sensitive, but require high-equipped laboratories and well-trained personnel. Hence, in this study, a rapid point-of-need detection method was developed to detect all Rickettsia species. The 23S and 16S rRNA genes were targeted to develop a recombinase polymerase amplification (RPA) assay. Both 23S and 16S_RPA assays required between seven to ten minutes to amplify and detect one or ten DNA molecules/reaction, respectively. The 16S_RPA assay detected all tested species, whereas the 23S_RPA assay identified only species of the spotted fever and transitional rickettsial groups. All results were compared with real-time PCR assays directed against the same rickettsial genes. The RPA assays are easy to handle and produced quicker results in comparison to real-time PCRs. Both RPA assays were implemented in a mobile suitcase laboratory to ease the use in rural areas. This method can help to provide rapid management of rickettsial infections. Copyright © 2017 Elsevier Inc. All rights reserved.

Electrochemical Measurement of the β-Galactosidase Reporter from Live Cells: A Comparison to the Miller Assay.

PubMed

Tschirhart, Tanya; Zhou, Xinyi Y; Ueda, Hana; Tsao, Chen-Yu; Kim, Eunkyoung; Payne, Gregory F; Bentley, William E

2016-01-15

In order to match our ability to conceive of and construct cells with enhanced function, we must concomitantly develop facile, real-time methods for elucidating performance. With these, new designs can be tested in silico and steps in construction incrementally validated. Electrochemical monitoring offers the above advantages largely because signal transduction stems from direct electron transfer, allowing for potentially quicker and more integrated measurements. One of the most common genetic reporters, β-galactosidase, can be measured both spectrophotometrically (Miller assay) and electrochemically. However, since the relationship between the two is not well understood, the electrochemical methods have not yet garnered the attention of biologists. With the aim of demonstrating the utility of an electrochemical measurement to the synthetic biology community, we created a genetic construct that interprets and reports (with β-galactosidase) on the concentration of the bacterial quorum sensing molecule autoinducer-2. In this work, we provide a correlation between electrochemical measurements and Miller Units. We show that the electrochemical assay works with both lysed and whole cells, allowing for the prediction of one from the other, and for continuous monitoring of cell response. We further present a conceptually simple and generalized mathematical model for cell-based β-galactosidase reporter systems that could aid in building and predicting a variety of synthetic biology constructs. This first-ever in-depth comparison and analysis aims to facilitate the use of electrochemical real-time monitoring in the field of synthetic biology as well as to facilitate the creation of constructs that can more easily communicate information to electronic systems.

Fuzzy support vector machines for adaptive Morse code recognition.

PubMed

Yang, Cheng-Hong; Jin, Li-Cheng; Chuang, Li-Yeh

2006-11-01

Morse code is now being harnessed for use in rehabilitation applications of augmentative-alternative communication and assistive technology, facilitating mobility, environmental control and adapted worksite access. In this paper, Morse code is selected as a communication adaptive device for persons who suffer from muscle atrophy, cerebral palsy or other severe handicaps. A stable typing rate is strictly required for Morse code to be effective as a communication tool. Therefore, an adaptive automatic recognition method with a high recognition rate is needed. The proposed system uses both fuzzy support vector machines and the variable-degree variable-step-size least-mean-square algorithm to achieve these objectives. We apply fuzzy memberships to each point, and provide different contributions to the decision learning function for support vector machines. Statistical analyses demonstrated that the proposed method elicited a higher recognition rate than other algorithms in the literature.

Vectorization with SIMD extensions speeds up reconstruction in electron tomography.

PubMed

Agulleiro, J I; Garzón, E M; García, I; Fernández, J J

2010-06-01

Electron tomography allows structural studies of cellular structures at molecular detail. Large 3D reconstructions are needed to meet the resolution requirements. The processing time to compute these large volumes may be considerable and so, high performance computing techniques have been used traditionally. This work presents a vector approach to tomographic reconstruction that relies on the exploitation of the SIMD extensions available in modern processors in combination to other single processor optimization techniques. This approach succeeds in producing full resolution tomograms with an important reduction in processing time, as evaluated with the most common reconstruction algorithms, namely WBP and SIRT. The main advantage stems from the fact that this approach is to be run on standard computers without the need of specialized hardware, which facilitates the development, use and management of programs. Future trends in processor design open excellent opportunities for vector processing with processor's SIMD extensions in the field of 3D electron microscopy.

Assessing the impact of climate change on vector-borne viruses in the EU through the elicitation of expert opinion.

PubMed

Gale, P; Brouwer, A; Ramnial, V; Kelly, L; Kosmider, R; Fooks, A R; Snary, E L

2010-02-01

Expert opinion was elicited to undertake a qualitative risk assessment to estimate the current and future risks to the European Union (EU) from five vector-borne viruses listed by the World Organization for Animal Health. It was predicted that climate change will increase the risk of incursions of African horse sickness virus (AHSV), Crimean-Congo haemorrhagic fever virus (CCHFV) and Rift Valley fever virus (RVFV) into the EU from other parts of the world, with African swine fever virus (ASFV) and West Nile virus (WNV) being less affected. Currently the predicted risks of incursion were lowest for RVFV and highest for ASFV. Risks of incursion were considered for six routes of entry (namely vectors, livestock, meat products, wildlife, pets and people). Climate change was predicted to increase the risk of incursion from entry of vectors for all five viruses to some degree, the strongest effects being predicted for AHSV, CCHFV and WNV. This work will facilitate identification of appropriate risk management options in relation to adaptations to climate change.

Construction of two Lactococcus lactis expression vectors combining the Gateway and the NIsin Controlled Expression systems.

PubMed

Douillard, François P; Mahony, Jennifer; Campanacci, Valérie; Cambillau, Christian; van Sinderen, Douwe

2011-09-01

Over the last 10 years, the NIsin Controlled Expression (NICE) system has been extensively used in the food-grade bacterium Lactococcus lactis subsp. cremoris to produce homologous and heterologous proteins for academic and biotechnological purposes. Although various L. lactis molecular tools have been developed, no expression vectors harboring the popular Gateway recombination system are currently available for this widely used cloning host. In this study, we constructed two expression vectors that combine the NICE and the Gateway recombination systems and we tested their applicability by recombining and over-expressing genes encoding structural proteins of lactococcal phages Tuc2009 and TP901-1. Over-expressed phage proteins were analyzed by immunoblotting and purified by His-tag affinity chromatography with protein productions yielding 2.8-3.7 mg/l of culture. This therefore is the first description of L. lactis NICE expression vectors which integrate the Gateway cloning technology and which are suitable for the production of sufficient amounts of proteins to facilitate subsequent structural and functional analyses. Copyright © 2011 Elsevier Inc. All rights reserved.

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

PubMed Central

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. PMID:26445723

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

PubMed

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.

A multicolor panel of TALE-KRAB based transcriptional repressor vectors enabling knockdown of multiple gene targets

PubMed Central

Zhang, Zhonghui; Wu, Elise; Qian, Zhijian; Wu, Wen-Shu

2014-01-01

Stable and efficient knockdown of multiple gene targets is highly desirable for dissection of molecular pathways. Because it allows sequence-specific DNA binding, transcription activator-like effector (TALE) offers a new genetic perturbation technique that allows for gene-specific repression. Here, we constructed a multicolor lentiviral TALE-Kruppel-associated box (KRAB) expression vector platform that enables knockdown of multiple gene targets. This platform is fully compatible with the Golden Gate TALEN and TAL Effector Kit 2.0, a widely used and efficient method for TALE assembly. We showed that this multicolor TALE-KRAB vector system when combined together with bone marrow transplantation could quickly knock down c-kit and PU.1 genes in hematopoietic stem and progenitor cells of recipient mice. Furthermore, our data demonstrated that this platform simultaneously knocked down both c-Kit and PU.1 genes in the same primary cell populations. Together, our results suggest that this multicolor TALE-KRAB vector platform is a promising and versatile tool for knockdown of multiple gene targets and could greatly facilitate dissection of molecular pathways. PMID:25475013

A multicolor panel of TALE-KRAB based transcriptional repressor vectors enabling knockdown of multiple gene targets.

PubMed

Zhang, Zhonghui; Wu, Elise; Qian, Zhijian; Wu, Wen-Shu

2014-12-05

Stable and efficient knockdown of multiple gene targets is highly desirable for dissection of molecular pathways. Because it allows sequence-specific DNA binding, transcription activator-like effector (TALE) offers a new genetic perturbation technique that allows for gene-specific repression. Here, we constructed a multicolor lentiviral TALE-Kruppel-associated box (KRAB) expression vector platform that enables knockdown of multiple gene targets. This platform is fully compatible with the Golden Gate TALEN and TAL Effector Kit 2.0, a widely used and efficient method for TALE assembly. We showed that this multicolor TALE-KRAB vector system when combined together with bone marrow transplantation could quickly knock down c-kit and PU.1 genes in hematopoietic stem and progenitor cells of recipient mice. Furthermore, our data demonstrated that this platform simultaneously knocked down both c-Kit and PU.1 genes in the same primary cell populations. Together, our results suggest that this multicolor TALE-KRAB vector platform is a promising and versatile tool for knockdown of multiple gene targets and could greatly facilitate dissection of molecular pathways.

Perceiving patterns of play in dynamic sport tasks: investigating the essential information underlying skilled performance.

PubMed

Willams, A Mark; Hodges, Nicola J; North, Jamie S; Barton, Gabor

2006-01-01

The perceptual-cognitive information used to support pattern-recognition skill in soccer was examined. In experiment 1, skilled players were quicker and more accurate than less-skilled players at recognising familiar and unfamiliar soccer action sequences presented on film. In experiment 2, these action sequences were converted into point-light displays, with superficial display features removed and the positions of players and the relational information between them made more salient. Skilled players were more accurate than less-skilled players in recognising sequences presented in point-light form, implying that each pattern of play can be defined by the unique relations between players. In experiment 3, various offensive and defensive players were occluded for the duration of each trial in an attempt to identify the most important sources of information underpinning successful performance. A decrease in response accuracy was observed under occluded compared with non-occluded conditions and the expertise effect was no longer observed. The relational information between certain key players, team-mates and their defensive counterparts may provide the essential information for effective pattern-recognition skill in soccer. Structural feature analysis, temporal phase relations, and knowledge-based information are effectively integrated to facilitate pattern recognition in dynamic sport tasks.

Delegation within municipal health care.

PubMed

Bystedt, Maria; Eriksson, Maria; Wilde-Larsson, Bodil

2011-05-01

To describe how registered nurses (RNs) perceive delegation to unlicensed personnel (UP) in a municipal healthcare context in Sweden. Within municipal health care RNs often delegate tasks to UP. The latter have practical training, but lack formal competence. Twelve RNs were interviewed and the material was analysed using a phenomenographic approach. Owing to a shortage of RNs, delegation is seen as a prerequisite for a functioning organization. This necessity also involves a number of perceived contradictions in three areas: (1) the work situation of RNs - facilitation and relief vs. lack of control, powerlessness, vagueness regarding responsibility, and resignation; (2) the relationship with unlicensed personnel - stimulation, possibility for mentoring, use of UP competence and the creation of fairness vs. questioning UP competence; and (3) The patients - increase in continuity, quicker treatment, and increased security vs. insecurity (with respect to, for example, the handling of medicine). Registered nurses perceptions of delegation within municipal healthcare involve their own work situation, the UP and the patients. Registered nurses who delegate to UP must be given time for mentoring such that the nursing care is safe care of high quality. © 2011 The Authors. Journal compilation © 2011 Blackwell Publishing Ltd.

78 FR 18268 - Proposed Establishment of Class E Airspace; Blue Mesa, CO

Federal Register 2010, 2011, 2012, 2013, 2014

2013-03-26

...-0193; Airspace Docket No. 13-ANM-9] Proposed Establishment of Class E Airspace; Blue Mesa, CO AGENCY... action proposes to establish Class E airspace at the Blue Mesa VHF Omni-Directional Radio Range/Distance Measuring Equipment (VOR/DME), Blue Mesa, CO to facilitate vectoring of Instrument Flight Rules (IFR...

78 FR 45474 - Proposed Establishment of Class E Airspace; Cut Bank, MT

Federal Register 2010, 2011, 2012, 2013, 2014

2013-07-29

...-0532; Airspace Docket No. 13-ANM-21] Proposed Establishment of Class E Airspace; Cut Bank, MT AGENCY... action proposes to establish Class E airspace at the Cut Bank VHF Omni-Directional Radio Range Tactical Air Navigational Aid (VORTAC) navigation aid, Cut Bank, MT, to facilitate vectoring of Instrument...

Behavioral responses of male Diaphorina citri (Hemiptera: Liviidae) to mating communication signals from vibration traps in citrus (Sapindales: Rutaceae) trees

USDA-ARS?s Scientific Manuscript database

The Asian citrus psyllid, Diaphorina citri Kuwayama, vectors the bacterium causing citrus greening disease, which has devastated citrus production worldwide wherever it has been introduced. To help monitor and target D citri populations in commercial groves, thereby facilitating more effective manag...

Hfq Regulates Biofilm Gut Blockage That Facilitates Flea-Borne Transmission of Yersinia pestis

PubMed Central

Rempe, Katherine A.; Hinz, Angela K.

2012-01-01

The plague bacillus Yersinia pestis can achieve transmission by biofilm blockage of the foregut proventriculus of its flea vector. Hfq is revealed to be essential for biofilm blockage formation and acquisition and fitness of Y. pestis during flea gut infection, consistent with posttranscriptional regulatory mechanisms in plague transmission. PMID:22328669

Baculoviral delivery of CRISPR/Cas9 facilitates efficient genome editing in human cells

PubMed Central

Hindriksen, Sanne; Bramer, Arne J.; Truong, My Anh; Vromans, Martijn J. M.; Post, Jasmin B.; Verlaan-Klink, Ingrid; Snippert, Hugo J.; Lens, Susanne M. A.

2017-01-01

The CRISPR/Cas9 system is a highly effective tool for genome editing. Key to robust genome editing is the efficient delivery of the CRISPR/Cas9 machinery. Viral delivery systems are efficient vehicles for the transduction of foreign genes but commonly used viral vectors suffer from a limited capacity in the genetic information they can carry. Baculovirus however is capable of carrying large exogenous DNA fragments. Here we investigate the use of baculoviral vectors as a delivery vehicle for CRISPR/Cas9 based genome-editing tools. We demonstrate transduction of a panel of cell lines with Cas9 and an sgRNA sequence, which results in efficient knockout of all four targeted subunits of the chromosomal passenger complex (CPC). We further show that introduction of a homology directed repair template into the same CRISPR/Cas9 baculovirus facilitates introduction of specific point mutations and endogenous gene tags. Tagging of the CPC recruitment factor Haspin with the fluorescent reporter YFP allowed us to study its native localization as well as recruitment to the cohesin subunit Pds5B. PMID:28640891

A versatile and efficient high-throughput cloning tool for structural biology.

PubMed

Geertsma, Eric R; Dutzler, Raimund

2011-04-19

Methods for the cloning of large numbers of open reading frames into expression vectors are of critical importance for challenging structural biology projects. Here we describe a system termed fragment exchange (FX) cloning that facilitates the high-throughput generation of expression constructs. The method is based on a class IIS restriction enzyme and negative selection markers. FX cloning combines attractive features of established recombination- and ligation-independent cloning methods: It allows the straightforward transfer of an open reading frame into a variety of expression vectors and is highly efficient and very economic in its use. In addition, FX cloning avoids the common but undesirable feature of significantly extending target open reading frames with cloning related sequences, as it leaves a minimal seam of only a single extra amino acid to either side of the protein. The method has proven to be very robust and suitable for all common pro- and eukaryotic expression systems. It considerably speeds up the generation of expression constructs compared to traditional methods and thus facilitates a broader expression screening.

Malaria in Kakuma refugee camp, Turkana, Kenya: facilitation of Anopheles arabiensis vector populations by installed water distribution and catchment systems

PubMed Central

2011-01-01

Background Malaria is a major health concern for displaced persons occupying refugee camps in sub-Saharan Africa, yet there is little information on the incidence of infection and nature of transmission in these settings. Kakuma Refugee Camp, located in a dry area of north-western Kenya, has hosted ca. 60,000 to 90,000 refugees since 1992, primarily from Sudan and Somalia. The purpose of this study was to investigate malaria prevalence and attack rate and sources of Anopheles vectors in Kakuma refugee camp, in 2005-2006, after a malaria epidemic was observed by staff at camp clinics. Methods Malaria prevalence and attack rate was estimated from cases of fever presenting to camp clinics and the hospital in August 2005, using rapid diagnostic tests and microscopy of blood smears. Larval habitats of vectors were sampled and mapped. Houses were sampled for adult vectors using the pyrethrum knockdown spray method, and mapped. Vectors were identified to species level and their infection with Plasmodium falciparum determined. Results Prevalence of febrile illness with P. falciparum was highest among the 5 to 17 year olds (62.4%) while malaria attack rate was highest among the two to 4 year olds (5.2/1,000/day). Infected individuals were spatially concentrated in three of the 11 residential zones of the camp. The indoor densities of Anopheles arabiensis, the sole malaria vector, were similar during the wet and dry seasons, but were distributed in an aggregated fashion and predominantly in the same zones where malaria attack rates were high. Larval habitats and larval populations were also concentrated in these zones. Larval habitats were man-made pits of water associated with tap-stands installed as the water delivery system to residents with year round availability in the camp. Three percent of A. arabiensis adult females were infected with P. falciparum sporozoites in the rainy season. Conclusions Malaria in Kakuma refugee camp was due mainly to infection with P. falciparum and showed a hyperendemic age-prevalence profile, in an area with otherwise low risk of malaria given prevailing climate. Transmission was sustained by A. arabiensis, whose populations were facilitated by installation of man-made water distribution and catchment systems. PMID:21639926

Tick-Pathogen Interactions and Vector Competence: Identification of Molecular Drivers for Tick-Borne Diseases

PubMed Central

de la Fuente, José; Antunes, Sandra; Bonnet, Sarah; Cabezas-Cruz, Alejandro; Domingos, Ana G.; Estrada-Peña, Agustín; Johnson, Nicholas; Kocan, Katherine M.; Mansfield, Karen L.; Nijhof, Ard M.; Papa, Anna; Rudenko, Nataliia; Villar, Margarita; Alberdi, Pilar; Torina, Alessandra; Ayllón, Nieves; Vancova, Marie; Golovchenko, Maryna; Grubhoffer, Libor; Caracappa, Santo; Fooks, Anthony R.; Gortazar, Christian; Rego, Ryan O. M.

2017-01-01

Ticks and the pathogens they transmit constitute a growing burden for human and animal health worldwide. Vector competence is a component of vectorial capacity and depends on genetic determinants affecting the ability of a vector to transmit a pathogen. These determinants affect traits such as tick-host-pathogen and susceptibility to pathogen infection. Therefore, the elucidation of the mechanisms involved in tick-pathogen interactions that affect vector competence is essential for the identification of molecular drivers for tick-borne diseases. In this review, we provide a comprehensive overview of tick-pathogen molecular interactions for bacteria, viruses, and protozoa affecting human and animal health. Additionally, the impact of tick microbiome on these interactions was considered. Results show that different pathogens evolved similar strategies such as manipulation of the immune response to infect vectors and facilitate multiplication and transmission. Furthermore, some of these strategies may be used by pathogens to infect both tick and mammalian hosts. Identification of interactions that promote tick survival, spread, and pathogen transmission provides the opportunity to disrupt these interactions and lead to a reduction in tick burden and the prevalence of tick-borne diseases. Targeting some of the similar mechanisms used by the pathogens for infection and transmission by ticks may assist in development of preventative strategies against multiple tick-borne diseases. PMID:28439499

Mosquito Oviposition Behavior and Vector Control

PubMed Central

Day, Jonathan F.

2016-01-01

The burden of gene transfer from one mosquito generation to the next falls on the female and her eggs. The selection of an oviposition site that guarantees egg and larval survival is a critical step in the reproductive process. The dangers associated with ephemeral aquatic habitats, lengthy droughts, freezing winters, and the absence of larval nutrition makes careful oviposition site selection by a female mosquito extremely important. Mosquito species exhibit a remarkable diversity of oviposition behaviors that ensure eggs are deposited into microenvironments conducive for successful larval development and the emergence of the next mosquito generation. An understanding of mosquito oviposition behavior is necessary for the development of surveillance and control opportunities directed against specific disease vectors. For example, Aedes aegypti Linnaeus is the vector of viruses causing important human diseases including yellow fever, dengue, chikungunya, and Zika. The preference of this species to oviposit in natural and artificial containers has facilitated the development of Ae. aegypti-specific surveillance and toxic oviposition traps designed to detect and control this important vector species in and around disease foci. A better understanding of the wide diversity of mosquito oviposition behavior will allow the development of new and innovative surveillance and control devices directed against other important mosquito vectors of human and animal disease. PMID:27869724

Improvement of a yeast self-excising integrative vector by prevention of expression leakage of the intronated Cre recombinase gene during plasmid maintenance in Escherichia coli.

PubMed

Agaphonov, Michael O

2017-12-01

The use of plasmids possessing a regulatable gene coding for a site-specific recombinase together with its recognition sequences significantly facilitates genome manipulations since it allows self-excision of the portion of the genetic construct integrated into the host genome. Stable maintenance of such plasmids in Escherichia coli, which is used for plasmid preparation, requires prevention of recombinase synthesis in this host, which can be achieved by interrupting the recombinase gene with an intron. Based on this approach, Saccharomyces cerevisiae and Hansenula polymorpha self-excising vectors possessing intronated gene for Cre recombinase and its recognition sites (LoxP) were previously constructed. However, this work shows instability of the H. polymorpha vectors during plasmid maintenance in E. coli cells. This could be due to recombination between the loxP sites caused by residual expression of the cre gene. Prevention of translation reinitiation on an internal methionine codon completely solved this problem. A similar modification was made in a self-excising vector designed for S. cerevisiae. Apart from substantial improvement of yeast self-excising vectors, the obtained results also narrow down the essential part of Cre sequence. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Technology in International Admissions

ERIC Educational Resources Information Center

White, Elizabeth

2012-01-01

In a relatively short time, technology applications have become an essential feature of the admissions business. They make the jobs of international admissions professionals easier in many ways, allowing for more robust communication with applicants and counselors, a streamlined application process, and quicker access to information about…

Systems Biology and Biomarkers of Early Effects for Occupational Exposure Limit Setting

EPA Science Inventory

AbstractIn a recent National Research Council document, new strategies for risk assessment were described to enable more accurate and quicker assessments (()(1)()). This report suggested that evaluating individual responses through increased use of biomonitoring could improve dos...

Moving Genetic Biomonitoring from a Concept to a Tool

EPA Science Inventory

Molecular genetic techniques like DNA barcoding and environmental DNA have been proposed as tools for aquatic biomonitoring for nearly a decade, but have yet to break through into widespread acceptance. The potential benefits of these methods, such as quicker, cheaper, more detai...

Open resource metagenomics: a model for sharing metagenomic libraries.

PubMed

Neufeld, J D; Engel, K; Cheng, J; Moreno-Hagelsieb, G; Rose, D R; Charles, T C

2011-11-30

Both sequence-based and activity-based exploitation of environmental DNA have provided unprecedented access to the genomic content of cultivated and uncultivated microorganisms. Although researchers deposit microbial strains in culture collections and DNA sequences in databases, activity-based metagenomic studies typically only publish sequences from the hits retrieved from specific screens. Physical metagenomic libraries, conceptually similar to entire sequence datasets, are usually not straightforward to obtain by interested parties subsequent to publication. In order to facilitate unrestricted distribution of metagenomic libraries, we propose the adoption of open resource metagenomics, in line with the trend towards open access publishing, and similar to culture- and mutant-strain collections that have been the backbone of traditional microbiology and microbial genetics. The concept of open resource metagenomics includes preparation of physical DNA libraries, preferably in versatile vectors that facilitate screening in a diversity of host organisms, and pooling of clones so that single aliquots containing complete libraries can be easily distributed upon request. Database deposition of associated metadata and sequence data for each library provides researchers with information to select the most appropriate libraries for further research projects. As a starting point, we have established the Canadian MetaMicroBiome Library (CM(2)BL [1]). The CM(2)BL is a publicly accessible collection of cosmid libraries containing environmental DNA from soils collected from across Canada, spanning multiple biomes. The libraries were constructed such that the cloned DNA can be easily transferred to Gateway® compliant vectors, facilitating functional screening in virtually any surrogate microbial host for which there are available plasmid vectors. The libraries, which we are placing in the public domain, will be distributed upon request without restriction to members of both the academic research community and industry. This article invites the scientific community to adopt this philosophy of open resource metagenomics to extend the utility of functional metagenomics beyond initial publication, circumventing the need to start from scratch with each new research project.

Open resource metagenomics: a model for sharing metagenomic libraries

PubMed Central

Neufeld, J.D.; Engel, K.; Cheng, J.; Moreno-Hagelsieb, G.; Rose, D.R.; Charles, T.C.

2011-01-01

Both sequence-based and activity-based exploitation of environmental DNA have provided unprecedented access to the genomic content of cultivated and uncultivated microorganisms. Although researchers deposit microbial strains in culture collections and DNA sequences in databases, activity-based metagenomic studies typically only publish sequences from the hits retrieved from specific screens. Physical metagenomic libraries, conceptually similar to entire sequence datasets, are usually not straightforward to obtain by interested parties subsequent to publication. In order to facilitate unrestricted distribution of metagenomic libraries, we propose the adoption of open resource metagenomics, in line with the trend towards open access publishing, and similar to culture- and mutant-strain collections that have been the backbone of traditional microbiology and microbial genetics. The concept of open resource metagenomics includes preparation of physical DNA libraries, preferably in versatile vectors that facilitate screening in a diversity of host organisms, and pooling of clones so that single aliquots containing complete libraries can be easily distributed upon request. Database deposition of associated metadata and sequence data for each library provides researchers with information to select the most appropriate libraries for further research projects. As a starting point, we have established the Canadian MetaMicroBiome Library (CM2BL [1]). The CM2BL is a publicly accessible collection of cosmid libraries containing environmental DNA from soils collected from across Canada, spanning multiple biomes. The libraries were constructed such that the cloned DNA can be easily transferred to Gateway® compliant vectors, facilitating functional screening in virtually any surrogate microbial host for which there are available plasmid vectors. The libraries, which we are placing in the public domain, will be distributed upon request without restriction to members of both the academic research community and industry. This article invites the scientific community to adopt this philosophy of open resource metagenomics to extend the utility of functional metagenomics beyond initial publication, circumventing the need to start from scratch with each new research project. PMID:22180823

Genomic resources for gene discovery, functional genome annotation, and evolutionary studies of maize and its close relatives.

PubMed

Wang, Chao; Shi, Xue; Liu, Lin; Li, Haiyan; Ammiraju, Jetty S S; Kudrna, David A; Xiong, Wentao; Wang, Hao; Dai, Zhaozhao; Zheng, Yonglian; Lai, Jinsheng; Jin, Weiwei; Messing, Joachim; Bennetzen, Jeffrey L; Wing, Rod A; Luo, Meizhong

2013-11-01

Maize is one of the most important food crops and a key model for genetics and developmental biology. A genetically anchored and high-quality draft genome sequence of maize inbred B73 has been obtained to serve as a reference sequence. To facilitate evolutionary studies in maize and its close relatives, much like the Oryza Map Alignment Project (OMAP) (www.OMAP.org) bacterial artificial chromosome (BAC) resource did for the rice community, we constructed BAC libraries for maize inbred lines Zheng58, Chang7-2, and Mo17 and maize wild relatives Zea mays ssp. parviglumis and Tripsacum dactyloides. Furthermore, to extend functional genomic studies to maize and sorghum, we also constructed binary BAC (BIBAC) libraries for the maize inbred B73 and the sorghum landrace Nengsi-1. The BAC/BIBAC vectors facilitate transfer of large intact DNA inserts from BAC clones to the BIBAC vector and functional complementation of large DNA fragments. These seven Zea Map Alignment Project (ZMAP) BAC/BIBAC libraries have average insert sizes ranging from 92 to 148 kb, organellar DNA from 0.17 to 2.3%, empty vector rates between 0.35 and 5.56%, and genome equivalents of 4.7- to 8.4-fold. The usefulness of the Parviglumis and Tripsacum BAC libraries was demonstrated by mapping clones to the reference genome. Novel genes and alleles present in these ZMAP libraries can now be used for functional complementation studies and positional or homology-based cloning of genes for translational genomics.

Facile manipulation of protein localization in fission yeast through binding of GFP-binding protein to GFP.

PubMed

Chen, Ying-Hui; Wang, Gao-Yuan; Hao, Hao-Chao; Chao, Chun-Jiang; Wang, Yamei; Jin, Quan-Wen

2017-03-01

GFP-binding protein (or GBP) has been recently developed in various systems and organisms as an efficient tool to purify GFP-fusion proteins. Due to the high affinity between GBP and GFP or GFP variants, this GBP-based approach is also ideally suited to alter the localization of functional proteins in live cells. In order to facilitate the wide use of the GBP-targeting approach in the fission yeast Schizosaccharomyces pombe , we developed a set of pFA6a-, pJK148- and pUC119-based vectors containing GBP- or GBP-mCherry-coding sequences and variants of inducible nmt1 or constitutive adh1 promoters that result in different levels of expression. The GBP or GBP-mCherry fragments can serve as cassettes for N- or C-terminal genomic tagging of genes of interest. We illustrated the application of these vectors in the construction of yeast strains with Dma1 or Cdc7 tagged with GBP-mCherry and efficient targeting of Dma1- or Cdc7-GBP-mCherry to the spindle pole body by Sid4-GFP. This series of vectors should help to facilitate the application of the GBP-targeting approach in manipulating protein localization and the analysis of gene function in fission yeast, at the level of single genes, as well as at a systematic scale. © 2017. Published by The Company of Biologists Ltd.

Better Together: Association With 'Candidatus Liberibacter Asiaticus' Increases the Reproductive Fitness of Its Insect Vector, Diaphorina citri (Hemiptera: Liviidae).

PubMed

Pelz-Stelinski, K S; Killiny, N

2016-05-01

The duration of the evolutionary association between a pathogen and vector can be inferred based on the strength of their mutualistic interactions. A well-adapted pathogen is likely to confer some benefit or, at a minimum, exhibit low pathogenicity toward its host vector. Coevolution of the two toward a mutually beneficial association appears to have occurred between the citrus greening disease pathogen, Candidatus Liberibacter asiaticus (Las), and its insect vector, the Asian citrus psyllid, Diaphorina citri (Kuwayama). To better understand the dynamics facilitating transmission, we evaluated the effects of Las infection on the fitness of its vector. Diaphorina citri harboring Las were more fecund than their uninfected counterparts; however, their nymphal development rate and adult survival were comparatively reduced. The finite rate of population increase and net reproductive rate were both greater among Las-infected D. citri as compared with uninfected counterparts, indicating that overall population fitness of infected psyllids was improved given the greater number of offspring produced. Previous reports of transovarial transmission, in conjunction with increased fecundity and population growth rates of Las-positive D. citri found in the current investigation, suggest a long evolutionary relationship between pathogen and vector. The survival of Las-infected adult D. citri was lower compared with uninfected D. citri , which suggests that there may be a fitness trade-off in response to Las infection. A beneficial effect of a plant pathogen on vector fitness may indicate that the pathogen developed a relationship with the insect before secondarily moving to plants.

Better Together: Association With ‘Candidatus Liberibacter Asiaticus’ Increases the Reproductive Fitness of Its Insect Vector, Diaphorina citri (Hemiptera: Liviidae)

PubMed Central

Pelz-Stelinski, K. S.; Killiny, N.

2016-01-01

The duration of the evolutionary association between a pathogen and vector can be inferred based on the strength of their mutualistic interactions. A well-adapted pathogen is likely to confer some benefit or, at a minimum, exhibit low pathogenicity toward its host vector. Coevolution of the two toward a mutually beneficial association appears to have occurred between the citrus greening disease pathogen, Candidatus Liberibacter asiaticus (Las), and its insect vector, the Asian citrus psyllid, Diaphorina citri (Kuwayama). To better understand the dynamics facilitating transmission, we evaluated the effects of Las infection on the fitness of its vector. Diaphorina citri harboring Las were more fecund than their uninfected counterparts; however, their nymphal development rate and adult survival were comparatively reduced. The finite rate of population increase and net reproductive rate were both greater among Las-infected D. citri as compared with uninfected counterparts, indicating that overall population fitness of infected psyllids was improved given the greater number of offspring produced. Previous reports of transovarial transmission, in conjunction with increased fecundity and population growth rates of Las-positive D. citri found in the current investigation, suggest a long evolutionary relationship between pathogen and vector. The survival of Las-infected adult D. citri was lower compared with uninfected D. citri, which suggests that there may be a fitness trade-off in response to Las infection. A beneficial effect of a plant pathogen on vector fitness may indicate that the pathogen developed a relationship with the insect before secondarily moving to plants. PMID:27418697

Improving protein–protein interactions prediction accuracy using protein evolutionary information and relevance vector machine model

PubMed Central

An, Ji‐Yong; Meng, Fan‐Rong; Chen, Xing; Yan, Gui‐Ying; Hu, Ji‐Pu

2016-01-01

Abstract Predicting protein–protein interactions (PPIs) is a challenging task and essential to construct the protein interaction networks, which is important for facilitating our understanding of the mechanisms of biological systems. Although a number of high‐throughput technologies have been proposed to predict PPIs, there are unavoidable shortcomings, including high cost, time intensity, and inherently high false positive rates. For these reasons, many computational methods have been proposed for predicting PPIs. However, the problem is still far from being solved. In this article, we propose a novel computational method called RVM‐BiGP that combines the relevance vector machine (RVM) model and Bi‐gram Probabilities (BiGP) for PPIs detection from protein sequences. The major improvement includes (1) Protein sequences are represented using the Bi‐gram probabilities (BiGP) feature representation on a Position Specific Scoring Matrix (PSSM), in which the protein evolutionary information is contained; (2) For reducing the influence of noise, the Principal Component Analysis (PCA) method is used to reduce the dimension of BiGP vector; (3) The powerful and robust Relevance Vector Machine (RVM) algorithm is used for classification. Five‐fold cross‐validation experiments executed on yeast and Helicobacter pylori datasets, which achieved very high accuracies of 94.57 and 90.57%, respectively. Experimental results are significantly better than previous methods. To further evaluate the proposed method, we compare it with the state‐of‐the‐art support vector machine (SVM) classifier on the yeast dataset. The experimental results demonstrate that our RVM‐BiGP method is significantly better than the SVM‐based method. In addition, we achieved 97.15% accuracy on imbalance yeast dataset, which is higher than that of balance yeast dataset. The promising experimental results show the efficiency and robust of the proposed method, which can be an automatic decision support tool for future proteomics research. For facilitating extensive studies for future proteomics research, we developed a freely available web server called RVM‐BiGP‐PPIs in Hypertext Preprocessor (PHP) for predicting PPIs. The web server including source code and the datasets are available at http://219.219.62.123:8888/BiGP/. PMID:27452983

Disclosing the Parameters Leading to High Productivity of Retroviral Producer Cells Lines: Evaluating Random Versus Targeted Integration.

PubMed

Bandeira, Vanessa S; Tomás, Hélio A; Alici, Evren; Carrondo, Manuel J T; Coroadinha, Ana S

2017-04-01

Gammaretrovirus and lentivirus are the preferred viral vectors to genetically modify T and natural killer cells to be used in immune cell therapies. The transduction efficiency of hematopoietic and T cells is more efficient using gibbon ape leukemia virus (GaLV) pseudotyping. In this context gammaretroviral vector producer cells offer competitive higher titers than transient lentiviral vectors productions. The main aim of this work was to identify the key parameters governing GaLV-pseudotyped gammaretroviral vector productivity in stable producer cells, using a retroviral vector expression cassette enabling positive (facilitating cell enrichment) and negative cell selection (allowing cell elimination). The retroviral vector contains a thymidine kinase suicide gene fused with a ouabain-resistant Na + ,K + -ATPase gene, a potential safer and faster marker. The establishment of retroviral vector producer cells is traditionally performed by randomly integrating the retroviral vector expression cassette codifying the transgene. More recently, recombinase-mediated cassette exchange methodologies have been introduced to achieve targeted integration. Herein we compared random and targeted integration of the retroviral vector transgene construct. Two retroviral producer cell lines, 293 OuaS and 293 FlexOuaS, were generated by random and targeted integration, respectively, producing high titers (on the order of 10 7 infectious particles·ml -1 ). Results showed that the retroviral vector transgene cassette is the key retroviral vector component determining the viral titers notwithstanding, single-copy integration is sufficient to provide high titers. The expression levels of the three retroviral constructs (gag-pol, GaLV env, and retroviral vector transgene) were analyzed. Although gag-pol and GaLV env gene expression levels should surpass a minimal threshold, we found that relatively modest expression levels of these two expression cassettes are required. Their levels of expression should not be maximized. We concluded, to establish a high producer retroviral vector cell line only the expression level of the genomic retroviral RNA, that is, the retroviral vector transgene cassette, should be maximized, both through (1) the optimization of its design (i.e., genetic elements composition) and (2) the selection of high expressing chromosomal locus for its integration. The use of methodologies identifying and promoting integration into high-expression loci, as targeted integration or high-throughput screening are in this perspective highly valuable.

Simple and Effective Algorithms: Computer-Adaptive Testing.

ERIC Educational Resources Information Center

Linacre, John Michael

Computer-adaptive testing (CAT) allows improved security, greater scoring accuracy, shorter testing periods, quicker availability of results, and reduced guessing and other undesirable test behavior. Simple approaches can be applied by the classroom teacher, or other content specialist, who possesses simple computer equipment and elementary…

Local area networking: Ames centerwide network

NASA Technical Reports Server (NTRS)

Price, Edwin

1988-01-01

A computer network can benefit the user by making his/her work quicker and easier. A computer network is made up of seven different layers with the lowest being the hardware, the top being the user, and the middle being the software. These layers are discussed.

Relatively Certain! Comparative Thinking Reduces Uncertainty

ERIC Educational Resources Information Center

Mussweiler, Thomas; Posten, Ann-Christin

2012-01-01

Comparison is one of the most ubiquitous and versatile mechanisms in human information processing. Previous research demonstrates that one consequence of comparative thinking is increased judgmental efficiency: Comparison allows for quicker judgments without a loss in accuracy. We hypothesised that a second potential consequence of comparative…

Thin plastic sheet eliminates need for expensive plating

NASA Technical Reports Server (NTRS)

Stremel, R. L.

1967-01-01

Gasket of a commercially available plastic material is interposed between the mating surfaces in axial joints where a hard and a soft metal are in intimate contact under stress conditions. This eliminates the fretting problem and is quicker and less expensive than the plating process.

Student Mobility & Credit Transfer: A National and Global Survey

ERIC Educational Resources Information Center

Junor, Sean; Usher, Alex

2008-01-01

Technological advances in communication and transportation have increased educational opportunities for post-secondary education students around the globe. Individuals and information now travel quicker and cheaper between countries and continents. Post-secondary institutions no longer have a local, jurisdictional or even domestic focus; their…

Wavelet Transform Analysis of the Power Spectrum of Centre of Pressure Signals to Detect the Critical Point Interval of Postural Control

NASA Astrophysics Data System (ADS)

Singh, Neeraj Kumar; Snoussi, Hichem; Hewson, David; Duchêne, Jacques

The aim of this study was to develop a method to detecting the critical point interval (CPI) when sensory feedback is used as part of a closed-loop postural control strategy. Postural balance was evaluated using centre of pressure (COP) displacements from a force plate for 17 control and 10 elderly subjects under eyes open, eyes closed, and vibration conditions. A modified local-maximum-modulus wavelet transform analysis using the power spectrum of COP signals was used to calculate CPI. Lower CPI values indicate increased closed-loop postural control with a quicker response to sensory input. Such a strategy requires greater energy expenditure due to the repeated muscular interventions to remain stable. The CPI for elderly occurred significantly quicker than for controls, indicating tighter control of posture. Similar results were observed for eyes closed and vibration conditions. The CPI parameter can be used to detect differences in postural control due to ageing.

A series of vectors to construct lacZ fusions for the study of gene expression in Schizosaccharomyces pombe.

PubMed

Lafuente, M J; Petit, T; Gancedo, C

1997-12-22

We have constructed a series of plasmids to facilitate the fusion of promoters with or without coding regions of genes of Schizosaccharomyces pombe to the lacZ gene of Escherichia coli. These vectors carry a multiple cloning region in which fission yeast DNA may be inserted in three different reading frames with respect to the coding region of lacZ. The plasmids were constructed with the ura4+ or the his3+ marker of S. pombe. Functionality of the plasmids was tested measuring in parallel the expression of fructose 1,6-bisphosphatase and beta-galactosidase under the control of the fbp1+ promoter in different conditions.

Cloning of a very virulent plus, 686 strain of Marek’s disease virus as a bacterial artificial chromosome

USDA-ARS?s Scientific Manuscript database

Bacterial artificial chromosome (BAC) vectors were first developed to facilitate propagation and manipulation of large DNA fragments. This technology was later used to clone full-length genomes of large DNA viruses to study viral gene function. Marek’s disease virus (MDV) is a highly oncogenic herpe...

78 FR 58158 - Establishment of Class E Airspace; Wasatch, UT

Federal Register 2010, 2011, 2012, 2013, 2014

2013-09-23

...., long. 111[deg]07'28'' W.; to Lat. 39[deg]03'55'' N., long. 110[deg]37'49'' W.; to Lat. 38[deg]28'51'' N... Aviation Administration (FAA), DOT. ACTION: Final rule. SUMMARY: This action establishes Class E airspace..., Wasatch, UT, to facilitate vectoring of Instrument Flight Rules (IFR) aircraft under control of Salt Lake...

Introduction of plasmid DNA into an ST398 livestock-associated methicillin-resistant Staphylococcus aureus strain

USDA-ARS?s Scientific Manuscript database

MRS926 is a livestock-associated methicillin-resistant Staphylococcus aureus (MRSA) strain of sequence type (ST) 398. In order to facilitate in vitro and in vivo studies of this strain, we sought to tag it with a fluorescent marker. We cloned a codon-optimized gene for TurboGFP into a shuttle vector...

Rule-based fuzzy vector median filters for 3D phase contrast MRI segmentation

NASA Astrophysics Data System (ADS)

Sundareswaran, Kartik S.; Frakes, David H.; Yoganathan, Ajit P.

2008-02-01

Recent technological advances have contributed to the advent of phase contrast magnetic resonance imaging (PCMRI) as standard practice in clinical environments. In particular, decreased scan times have made using the modality more feasible. PCMRI is now a common tool for flow quantification, and for more complex vector field analyses that target the early detection of problematic flow conditions. Segmentation is one component of this type of application that can impact the accuracy of the final product dramatically. Vascular segmentation, in general, is a long-standing problem that has received significant attention. Segmentation in the context of PCMRI data, however, has been explored less and can benefit from object-based image processing techniques that incorporate fluids specific information. Here we present a fuzzy rule-based adaptive vector median filtering (FAVMF) algorithm that in combination with active contour modeling facilitates high-quality PCMRI segmentation while mitigating the effects of noise. The FAVMF technique was tested on 111 synthetically generated PC MRI slices and on 15 patients with congenital heart disease. The results were compared to other multi-dimensional filters namely the adaptive vector median filter, the adaptive vector directional filter, and the scalar low pass filter commonly used in PC MRI applications. FAVMF significantly outperformed the standard filtering methods (p < 0.0001). Two conclusions can be drawn from these results: a) Filtering should be performed after vessel segmentation of PC MRI; b) Vector based filtering methods should be used instead of scalar techniques.

Tetrahedral DNA Nanoparticle Vector for Intracellular Delivery of Targeted Peptide Nucleic Acid Antisense Agents to Restore Antibiotic Sensitivity in Cefotaxime-Resistant Escherichia coli.

PubMed

Readman, John Benedict; Dickson, George; Coldham, Nick G

2017-06-01

The bacterial cell wall presents a barrier to the uptake of unmodified synthetic antisense oligonucleotides, such as peptide nucleic acids, and so is one of the greatest obstacles to the development of their use as therapeutic anti-bacterial agents. Cell-penetrating peptides have been covalently attached to antisense agents, to facilitate penetration of the bacterial cell wall and deliver their cargo into the cytoplasm. Although they are an effective vector for antisense oligonucleotides, they are not specific for bacterial cells and can exhibit growth inhibitory properties at higher doses. Using a bacterial cell growth assay in the presence of cefotaxime (CTX 16 mg/L), we have developed and evaluated a self-assembling non-toxic DNA tetrahedron nanoparticle vector incorporating a targeted anti-bla CTX-M-group 1 antisense peptide nucleic acid (PNA4) in its structure for penetration of the bacterial cell wall. A dose-dependent CTX potentiating effect was observed when PNA4 (0-40 μM) was incorporated into the structure of a DNA tetrahedron vector. The minimum inhibitory concentration (to CTX) of an Escherichia coli field isolate harboring a plasmid carrying bla CTX-M-3 was reduced from 35 to 16 mg/L in the presence of PNA4 carried by the DNA tetrahedron vector (40 μM), contrasting with no reduction in MIC in the presence of PNA4 alone. No growth inhibitory effects of the DNA tetrahedron vector alone were observed.

Implication of vector characteristics of Phlebotomus argentipes in the kala-azar elimination programme in the Indian sub-continent.

PubMed

Chowdhury, Rajib; Kumar, Vijay; Mondal, Dinesh; Das, Murari Lal; Das, Pradeep; Dash, Aditya Prasad; Kroeger, Axel

2016-05-01

Visceral leishmaniasis (VL), also known as kala-azar in the Indian sub-continent (ISC), is a major public health concern in Bangladesh, India, and Nepal, where it is caused by Leishmania donovani transmitted by the sand fly Phlebotomus argentipes. Various ecological parameters including air temperature, rainfall, wind speed, relative humidity, soil moisture, pH, and organic carbon are known to influence the oviposition of female sand flies, as well as the survival and development of larvae. However, more detailed knowledge on vector behavior, such as biting times, breeding places, and preferred hosts are needed to design optimal evidence-based vector control interventions. In order to facilitate rational decisions regarding VL vector control, a systematic review was conducted to identify the prevailing practice and knowledge gaps in relation to vector bionomics and behavior. Search terms included 'sand fly bionomics', 'habitat', and 'visceral leishmaniasis/kala-azar vector control' using the Boolean operator AND to identify the country of interest, namely: Bangladesh, India, and Nepal. Both PubMed and Google search engines were used. Additional unpublished documents in the three countries were also analyzed. Information on the life cycle of VL vectors, their breeding behavior, infection rate with L. donovani, feeding behavior, and seasonal variation are useful for designing vector control operations. Unfortunately, none of the studies on the life cycle of P. argentipes was conducted in field settings of the ISC, so the publications from other locations had to be used for determining the duration of life cycle and development from egg to adult. However, information about breeding places, seasonal variation of vector densities, and 47 out of the selected 51 papers are available from the ISC and can be used for intelligent design of control operations. Vector control services should undertake routine insecticide resistance monitoring and adapt indoor residual spraying rounds to the seasonality of vector densities. Further research is needed on potential animal reservoirs for L. donovani, on the breeding habitat, and life cycle of sand flies in the ISC.

Implication of vector characteristics of Phlebotomus argentipes in the kala-azar elimination programme in the Indian sub-continent

PubMed Central

Chowdhury, Rajib; Kumar, Vijay; Mondal, Dinesh; Das, Murari Lal; Das, Pradeep; Dash, Aditya Prasad

2016-01-01

Background Visceral leishmaniasis (VL), also known as kala-azar in the Indian sub-continent (ISC), is a major public health concern in Bangladesh, India, and Nepal, where it is caused by Leishmania donovani transmitted by the sand fly Phlebotomus argentipes. Various ecological parameters including air temperature, rainfall, wind speed, relative humidity, soil moisture, pH, and organic carbon are known to influence the oviposition of female sand flies, as well as the survival and development of larvae. However, more detailed knowledge on vector behavior, such as biting times, breeding places, and preferred hosts are needed to design optimal evidence-based vector control interventions. Methods In order to facilitate rational decisions regarding VL vector control, a systematic review was conducted to identify the prevailing practice and knowledge gaps in relation to vector bionomics and behavior. Search terms included ‘sand fly bionomics’, ‘habitat’, and ‘visceral leishmaniasis/kala-azar vector control’ using the Boolean operator AND to identify the country of interest, namely: Bangladesh, India, and Nepal. Both PubMed and Google search engines were used. Additional unpublished documents in the three countries were also analyzed. Results Information on the life cycle of VL vectors, their breeding behavior, infection rate with L. donovani, feeding behavior, and seasonal variation are useful for designing vector control operations. Unfortunately, none of the studies on the life cycle of P. argentipes was conducted in field settings of the ISC, so the publications from other locations had to be used for determining the duration of life cycle and development from egg to adult. However, information about breeding places, seasonal variation of vector densities, and 47 out of the selected 51 papers are available from the ISC and can be used for intelligent design of control operations. Conclusion Vector control services should undertake routine insecticide resistance monitoring and adapt indoor residual spraying rounds to the seasonality of vector densities. Further research is needed on potential animal reservoirs for L. donovani, on the breeding habitat, and life cycle of sand flies in the ISC. PMID:27376500

Strengthening tactical planning and operational frameworks for vector control: the roadmap for malaria elimination in Namibia.

PubMed

Chanda, Emmanuel; Ameneshewa, Birkinesh; Angula, Hans A; Iitula, Iitula; Uusiku, Pentrina; Trune, Desta; Islam, Quazi M; Govere, John M

2015-08-05

Namibia has made tremendous gains in malaria control and the epidemiological trend of the disease has changed significantly over the past years. In 2010, the country reoriented from the objective of reducing disease morbidity and mortality to the goal of achieving malaria elimination by 2020. This manuscript outlines the processes undertaken in strengthening tactical planning and operational frameworks for vector control to facilitate expeditious malaria elimination in Namibia. The information sources for this study included all available data and accessible archived documentary records on malaria vector control in Namibia. A methodical assessment of published and unpublished documents was conducted via a literature search of online electronic databases, Google Scholar, PubMed and WHO, using a combination of search terms. To attain the goal of elimination in Namibia, systems are being strengthened to identify and clear all infections, and significantly reduce human-mosquito contact. Particularly, consolidating vector control for reducing transmission at the identified malaria foci will be critical for accelerated malaria elimination. Thus, guarding against potential challenges and the need for evidence-based and sustainable vector control instigated the strengthening of strategic frameworks by: adopting the integrated vector management (IVM) strategy; initiating implementation of the global plan for insecticide resistance management (GPIRM); intensifying malaria vector surveillance; improving data collection and reporting systems on DDT; updating the indoor residual spraying (IRS) data collection and reporting tool; and, improving geographical reconnaissance using geographical information system-based satellite imagery. Universal coverage with IRS and long-lasting insecticidal nets, supplemented by larval source management in the context of IVM and guided by vector surveillance coupled with rational operationalization of the GPIRM, will enable expeditious attainment of elimination in Namibia. However, national capacity to plan, implement, monitor and evaluate interventions will require adequate and sustained support for technical, physical infrastructure, and human and financial resources for entomology and vector control operations.

How effective is integrated vector management against malaria and lymphatic filariasis where the diseases are transmitted by the same vector?

PubMed

Stone, Christopher M; Lindsay, Steve W; Chitnis, Nakul

2014-12-01

The opportunity to integrate vector management across multiple vector-borne diseases is particularly plausible for malaria and lymphatic filariasis (LF) control where both diseases are transmitted by the same vector. To date most examples of integrated control targeting these diseases have been unanticipated consequences of malaria vector control, rather than planned strategies that aim to maximize the efficacy and take the complex ecological and biological interactions between the two diseases into account. We developed a general model of malaria and LF transmission and derived expressions for the basic reproductive number (R0) for each disease. Transmission of both diseases was most sensitive to vector mortality and biting rate. Simulating different levels of coverage of long lasting-insecticidal nets (LLINs) and larval control confirms the effectiveness of these interventions for the control of both diseases. When LF was maintained near the critical density of mosquitoes, minor levels of vector control (8% coverage of LLINs or treatment of 20% of larval sites) were sufficient to eliminate the disease. Malaria had a far greater R0 and required a 90% population coverage of LLINs in order to eliminate it. When the mosquito density was doubled, 36% and 58% coverage of LLINs and larval control, respectively, were required for LF elimination; and malaria elimination was possible with a combined coverage of 78% of LLINs and larval control. Despite the low level of vector control required to eliminate LF, simulations suggest that prevalence of LF will decrease at a slower rate than malaria, even at high levels of coverage. If representative of field situations, integrated management should take into account not only how malaria control can facilitate filariasis elimination, but strike a balance between the high levels of coverage of (multiple) interventions required for malaria with the long duration predicted to be required for filariasis elimination.

Attraction Toward the Model and Model's Competence as Determinants of Adult Imitative Behavior

ERIC Educational Resources Information Center

Baron, Robert A.

1970-01-01

Suggests that adults are quicker to learn to match the performance of a model similar to themselves in attitude if he is competent. Similarity of the model interferes with rate of learning if he is incompetent. Tables, graph, and bibliography. (RW)

An Assessment of Differential Reinforcement Procedures for Learners with Autism Spectrum Disorder

ERIC Educational Resources Information Center

Johnson, Kate A.; Vladescu, Jason C.; Kodak, Tiffany; Sidener, Tina M.

2017-01-01

Differential reinforcement procedures may promote unprompted correct responding, resulting in a quicker transfer of stimulus control than nondifferential reinforcement. Recent studies that have compared reinforcement arrangements have found that the most effective arrangement may differ across participants. The current study conducted an…

Flavivirus NS1 protein in infected host sera enhances viral acquisition by mosquitoes.

PubMed

Liu, Jianying; Liu, Yang; Nie, Kaixiao; Du, Senyan; Qiu, Jingjun; Pang, Xiaojing; Wang, Penghua; Cheng, Gong

2016-06-20

The arbovirus life cycle involves viral transfer between a vertebrate host and an arthropod vector, and acquisition of virus from an infected mammalian host by a vector is an essential step in this process. Here, we report that flavivirus nonstructural protein-1 (NS1), which is abundantly secreted into the serum of an infected host, plays a critical role in flavivirus acquisition by mosquitoes. The presence of dengue virus (DENV) and Japanese encephalitis virus NS1s in the blood of infected interferon-α and γ receptor-deficient mice (AG6) facilitated virus acquisition by their native mosquito vectors because the protein enabled the virus to overcome the immune barrier of the mosquito midgut. Active immunization of AG6 mice with a modified DENV NS1 reduced DENV acquisition by mosquitoes and protected mice against a lethal DENV challenge, suggesting that immunization with NS1 could reduce the number of virus-carrying mosquitoes as well as the incidence of flaviviral diseases. Our study demonstrates that flaviviruses utilize NS1 proteins produced during their vertebrate phases to enhance their acquisition by vectors, which might be a result of flavivirus evolution to adapt to multiple host environments.

Flavivirus NS1 protein in infected host sera enhances viral acquisition by mosquitoes

PubMed Central

Liu, Jianying; Liu, Yang; Nie, Kaixiao; Du, Senyan; Qiu, Jingjun; Pang, Xiaojing; Wang, Penghua; Cheng, Gong

2016-01-01

Summary The arbovirus life cycle involves viral transfer between a vertebrate host and an arthropod vector, and acquisition of virus from an infected mammalian host by a vector is an essential step in this process. Here, we report that flavivirus nonstructural protein-1 (NS1), which is abundantly secreted into the serum of an infected host, plays a critical role in flavivirus acquisition by mosquitoes. The presence of dengue virus (DENV) and Japanese encephalitis virus (JEV) NS1s in the blood of infected interferon alpha and gamma receptor-deficient mice (AG6) facilitated virus acquisition by their native mosquito vectors because the protein enabled the virus to overcome the immune barrier of the mosquito midgut. Active immunization of AG6 mice with a modified DENV NS1 reduced DENV acquisition by mosquitoes and protected mice against a lethal DENV challenge, suggesting that immunization with NS1 could reduce the number of virus-carrying mosquitoes as well as the incidence of flaviviral diseases. Our study demonstrates that flaviviruses utilize NS1 proteins produced during their vertebrate phases to enhance their acquisition by vectors, which might be a result of flavivirus evolution to adapt to multiple host environments. PMID:27562253

Influence of molecular weight upon mannosylated bio-synthetic hybrids for targeted antigen presenting cell gene delivery

PubMed Central

Jones, Charles H.; Gollakota, Akhila; Chen, Mingfu; Chung, Tai-Chun; Ravikrishnan, Anitha; Zhang, Guojian; Pfeifer, Blaine A.

2015-01-01

Given the rise of antibiotic resistant microbes, genetic vaccination is a promising prophylactic strategy that enables rapid design and manufacture. Facilitating this process is the choice of vector, which is often situationally-specific and limited in engineering capacity. Furthermore, these shortcomings are usually tied to an incomplete understanding of the structure-function relationships driving vector-mediated gene delivery. Building upon our initial report of a hybrid bacterial-biomaterial gene delivery vector, a comprehensive structure-function assessment was completed using a class of mannosylated poly(beta-amino esters). Through a top-down screening methodology, an ideal polymer was selected on the basis of gene delivery efficacy and then used for the synthesis of a stratified molecular weight polymer library. By eliminating contributions of polymer chemical background, we were able to complete an in-depth assessment of gene delivery as a function of (1) polymer molecular weight, (2) relative mannose content, (3) polymer-membrane biophysical properties, (4) APC uptake specificity, and (5) serum inhibition. In summary, the flexibility and potential of the hybrid design featured in this work highlights the ability to systematically probe vector-associated properties for the development of translational gene delivery candidates. PMID:25941787

Rapid production of functionalized recombinant proteins: marrying ligation independent cloning and in vitro protein ligation.

PubMed

Kushnir, Susanna; Marsac, Yoann; Breitling, Reinhard; Granovsky, Igor; Brok-Volchanskaya, Vera; Goody, Roger S; Becker, Christian F W; Alexandrov, Kirill

2006-01-01

Functional genomics and proteomics have been very active fields since the sequencing of several genomes was completed. To assign a physiological role to the newly discovered coding genes with unknown function, new generic methods for protein production, purification, and targeted functionalization are needed. This work presents a new vector, pCYSLIC, that allows rapid generation of Escherichia coli expression constructs via ligation-independent cloning (LIC). The vector is designed to facilitate protein purification by either Ni-NTA or GSH affinity chromatography. Subsequent proteolytic removal of affinity tags liberates an N-terminal cysteine residue that is then used for covalent modification of the target protein with different biophysical probes via protein ligation. The described system has been tested on 36 mammalian Rab GTPases, and it was demonstrated that recombinant GTPases produced with pCYSLIC could be efficiently modified with fluorescein or biotin in vitro. Finally, LIC was compared with the recently developed In-Fusion cloning method, and it was demonstrated that In-Fusion provides superior flexibility in choice of expression vector. By the application of In-Fusion cloning Cys-Rab6A GTPase with an N-terminal cysteine residue was generated employing unmodified pET30a vector and TVMV protease.

The C Terminus of the Polerovirus P5 Readthrough Domain Limits Virus Infection to the Phloem▿

PubMed Central

Peter, Kari A.; Gildow, Frederick; Palukaitis, Peter; Gray, Stewart M.

2009-01-01

Poleroviruses are restricted to vascular phloem tissues from which they are transmitted by their aphid vectors and are not transmissible mechanically. Phloem limitation has been attributed to the absence of virus proteins either facilitating movement or counteracting plant defense. The polerovirus capsid is composed of two forms of coat protein, the major P3 protein and the minor P3/P5 protein, a translational readthrough of P3. P3/P5 is required for insect transmission and acts in trans to facilitate long-distance virus movement in phloem tissue. Specific potato leafroll virus mutants lacking part or all of the P5 domain moved into and infected nonvascular mesophyll tissue when the source-sink relationship of the plant (Solanum sarrachoides) was altered by pruning, with the progeny virus now being transmissible mechanically. However, in a period of months, a phloem-specific distribution of the virus was reestablished in the absence of aphid transmission. Virus from the new phloem-limited infection showed compensatory mutations that would be expected to restore the production of full-length P3/P5 as well as the loss of mechanical transmissibility. The data support our hypothesis that phloem limitation in poleroviruses presumably does not result from a deficiency in the repertoire of virus genes but rather results from P3/P5 accumulation under selection in the infected plant, with the colateral effect of facilitating transmission by phloem-feeding aphid vectors. PMID:19297484

The C terminus of the polerovirus p5 readthrough domain limits virus infection to the phloem.

PubMed

Peter, Kari A; Gildow, Frederick; Palukaitis, Peter; Gray, Stewart M

2009-06-01

Poleroviruses are restricted to vascular phloem tissues from which they are transmitted by their aphid vectors and are not transmissible mechanically. Phloem limitation has been attributed to the absence of virus proteins either facilitating movement or counteracting plant defense. The polerovirus capsid is composed of two forms of coat protein, the major P3 protein and the minor P3/P5 protein, a translational readthrough of P3. P3/P5 is required for insect transmission and acts in trans to facilitate long-distance virus movement in phloem tissue. Specific potato leafroll virus mutants lacking part or all of the P5 domain moved into and infected nonvascular mesophyll tissue when the source-sink relationship of the plant (Solanum sarrachoides) was altered by pruning, with the progeny virus now being transmissible mechanically. However, in a period of months, a phloem-specific distribution of the virus was reestablished in the absence of aphid transmission. Virus from the new phloem-limited infection showed compensatory mutations that would be expected to restore the production of full-length P3/P5 as well as the loss of mechanical transmissibility. The data support our hypothesis that phloem limitation in poleroviruses presumably does not result from a deficiency in the repertoire of virus genes but rather results from P3/P5 accumulation under selection in the infected plant, with the colateral effect of facilitating transmission by phloem-feeding aphid vectors.

Targeted Screening Strategies to Detect Trypanosoma cruzi Infection in Children

PubMed Central

Levy, Michael Z.; Kawai, Vivian; Bowman, Natalie M.; Waller, Lance A.; Cabrera, Lilia; Pinedo-Cancino, Viviana V.; Seitz, Amy E.; Steurer, Frank J.; Cornejo del Carpio, Juan G.; Cordova-Benzaquen, Eleazar; Maguire, James H.; Gilman, Robert H.; Bern, Caryn

2007-01-01

Background Millions of people are infected with Trypanosoma cruzi, the causative agent of Chagas disease in Latin America. Anti-trypanosomal drug therapy can cure infected individuals, but treatment efficacy is highest early in infection. Vector control campaigns disrupt transmission of T. cruzi, but without timely diagnosis, children infected prior to vector control often miss the window of opportunity for effective chemotherapy. Methods and Findings We performed a serological survey in children 2–18 years old living in a peri-urban community of Arequipa, Peru, and linked the results to entomologic, spatial and census data gathered during a vector control campaign. 23 of 433 (5.3% [95% CI 3.4–7.9]) children were confirmed seropositive for T. cruzi infection by two methods. Spatial analysis revealed that households with infected children were very tightly clustered within looser clusters of households with parasite-infected vectors. Bayesian hierarchical mixed models, which controlled for clustering of infection, showed that a child's risk of being seropositive increased by 20% per year of age and 4% per vector captured within the child's house. Receiver operator characteristic (ROC) plots of best-fit models suggest that more than 83% of infected children could be identified while testing only 22% of eligible children. Conclusions We found evidence of spatially-focal vector-borne T. cruzi transmission in peri-urban Arequipa. Ongoing vector control campaigns, in addition to preventing further parasite transmission, facilitate the collection of data essential to identifying children at high risk of T. cruzi infection. Targeted screening strategies could make integration of diagnosis and treatment of children into Chagas disease control programs feasible in lower-resource settings. PMID:18160979

Human Bocavirus Type-1 Capsid Facilitates the Transduction of Ferret Airways by Adeno-Associated Virus Genomes.

PubMed

Yan, Ziying; Feng, Zehua; Sun, Xingshen; Zhang, Yulong; Zou, Wei; Wang, Zekun; Jensen-Cody, Chandler; Liang, Bo; Park, Soo-Yeun; Qiu, Jianming; Engelhardt, John F

2017-08-01

Human bocavirus type-1 (HBoV1) has a high tropism for the apical membrane of human airway epithelia. The packaging of a recombinant adeno-associated virus 2 (rAAV2) genome into HBoV1 capsid produces a chimeric vector (rAAV2/HBoV1) that also efficiently transduces human airway epithelia. As such, this vector is attractive for use in gene therapies to treat lung diseases such as cystic fibrosis. However, preclinical development of rAAV2/HBoV1 vectors has been hindered by the fact that humans are the only known host for HBoV1 infection. This study reports that rAAV2/HBoV1 vector is capable of efficiently transducing the lungs of both newborn (3- to 7-day-old) and juvenile (29-day-old) ferrets, predominantly in the distal airways. Analyses of in vivo, ex vivo, and in vitro models of the ferret proximal airway demonstrate that infection of this particular region is less effective than it is in humans. Studies of vector binding and endocytosis in polarized ferret proximal airway epithelial cultures revealed that a lack of effective vector endocytosis is the main cause of inefficient transduction in vitro. While transgene expression declined proportionally with growth of the ferrets following infection at 7 days of age, reinfection of ferrets with rAAV2/HBoV1 at 29 days gave rise to approximately 5-fold higher levels of transduction than observed in naive infected 29-day-old animals. The findings presented here lay the foundation for clinical development of HBoV1 capsid-based vectors for lung gene therapy in cystic fibrosis using ferret models.

Water-soluble polymers bearing phosphorylcholine group and other zwitterionic groups for carrying DNA derivatives.

PubMed

Lin, Xiaojie; Ishihara, Kazuhiko

2014-01-01

Water-soluble polymers with equal positive and negative charges in the same monomer unit, such as the phosphorylcholine group and other zwitterionic groups, exhibit promising potential in gene delivery with appreciable transfection efficiency, compared with the traditional poly(ethylene glycol)-based polycation-gene complexes. These zwitterionic polymers with various architectural structures and properties have been synthesized by various polymerization methods, such as conventional radical polymerization, atom-transfer radical-polymerization, reversible addition-fragmentation chain-transfer polymerization, and nitroxide-mediated radical polymerization. These techniques have been used to efficiently facilitate gene therapy by fabrication of non-viral vectors with high cytocompatibility, large gene-carrying capacity, effective cell-membrane permeability, and in vivo gene-loading/releasing functionality. Zwitterionic polymer-based gene delivery vectors systems can be categorized into soluble-polymer/gene mixing, molecular self-assembly, and polymer-gene conjugation systems. This review describes the preparation and characterization of various zwitterionic polymer-based gene delivery vectors, specifically water-soluble phospholipid polymers for carrying gene derivatives.

The Gene Therapy Resource Program: A Decade of Dedication to Translational Research by the National Heart, Lung, and Blood Institute.

PubMed

Flotte, Terence R; Daniels, Eric; Benson, Janet; Bevett-Rose, Jeneé M; Cornetta, Kenneth; Diggins, Margaret; Johnston, Julie; Sepelak, Susan; van der Loo, Johannes C M; Wilson, James M; McDonald, Cheryl L

2017-12-01

Over a 10-year period, the Gene Therapy Resource Program (GTRP) of the National Heart Lung and Blood Institute has provided a set of core services to investigators to facilitate the clinical translation of gene therapy. These services have included a preclinical (research-grade) vector production core; current Good Manufacturing Practice clinical-grade vector cores for recombinant adeno-associated virus and lentivirus vectors; a pharmacology and toxicology core; and a coordinating center to manage program logistics and to provide regulatory and financial support to early-phase clinical trials. In addition, the GTRP has utilized a Steering Committee and a Scientific Review Board to guide overall progress and effectiveness and to evaluate individual proposals. These resources have been deployed to assist 82 investigators with 172 approved service proposals. These efforts have assisted in clinical trial implementation across a wide range of genetic, cardiac, pulmonary, and blood diseases. Program outcomes and potential future directions of the program are discussed.

CRISPR-Cas9 vectors for genome editing and host engineering in the baculovirus-insect cell system.

PubMed

Mabashi-Asazuma, Hideaki; Jarvis, Donald L

2017-08-22

The baculovirus-insect cell system (BICS) has been widely used to produce many different recombinant proteins for basic research and is being used to produce several biologics approved for use in human or veterinary medicine. Early BICS were technically complex and constrained by the relatively primordial nature of insect cell protein glycosylation pathways. Since then, recombination has been used to modify baculovirus vectors-which has simplified the system-and transform insect cells, which has enhanced its protein glycosylation capabilities. Now, CRISPR-Cas9 tools for site-specific genome editing are needed to facilitate further improvements in the BICS. Thus, in this study, we used various insect U6 promoters to construct CRISPR-Cas9 vectors and assessed their utility for site-specific genome editing in two insect cell lines commonly used as hosts in the BICS. We demonstrate the use of CRISPR-Cas9 to edit an endogenous insect cell gene and alter protein glycosylation in the BICS.

THE DEVELOPMENT AND TREND OF INSECTICIDE-RESISTANCE IN ANOPHELES ACONITUS DOENITZ AND ANOPHELES SUNDAICUS RODENWALDT.

PubMed

SOERONO, M; DAVIDSON, G; MUIR, D A

1965-01-01

The spread of insecticide-resistance in the two main vectors of malaria in Java, Anopheles aconitus Dönitz and A. sundaicus Rodenwaldt, has been of concern to workers in the Indonesian malaria eradication programme since DDT-resistance was first detected, in A. sundaicus, in 1954. The history of development of resistance in these two vectors has been summarized in this paper by assembling the available records in chronological order, thus facilitating reference.The picture which emerges is that of somewhat erratic occurrence of DDT-resistance in A. sundaicus and more stable, expanding areas of DDT-resistant A. aconitus. Dieldrin-resistance is more widespread than DDT-resistance in both species and preceded the onset of DDT-resistance in A. aconitus but not in A. sundaicus. These features should not be regarded as species characteristics, however, since selection pressures on the two species have been far from uniform.Malathion appears to be a promising alternative to DDT and dieldrin for the control of the two vectors in the circumstances described.

Genetic assignment methods for gaining insight into the management of infectious disease by understanding pathogen, vector, and host movement.

PubMed

Remais, Justin V; Xiao, Ning; Akullian, Adam; Qiu, Dongchuan; Blair, David

2011-04-01

For many pathogens with environmental stages, or those carried by vectors or intermediate hosts, disease transmission is strongly influenced by pathogen, host, and vector movements across complex landscapes, and thus quantitative measures of movement rate and direction can reveal new opportunities for disease management and intervention. Genetic assignment methods are a set of powerful statistical approaches useful for establishing population membership of individuals. Recent theoretical improvements allow these techniques to be used to cost-effectively estimate the magnitude and direction of key movements in infectious disease systems, revealing important ecological and environmental features that facilitate or limit transmission. Here, we review the theory, statistical framework, and molecular markers that underlie assignment methods, and we critically examine recent applications of assignment tests in infectious disease epidemiology. Research directions that capitalize on use of the techniques are discussed, focusing on key parameters needing study for improved understanding of patterns of disease.

Bartonella quintana Deploys Host and Vector Temperature-Specific Transcriptomes

PubMed Central

Previte, Domenic; Yoon, Kyong S.; Clark, J. Marshall; DeRisi, Joseph L.; Koehler, Jane E.

2013-01-01

The bacterial pathogen Bartonella quintana is passed between humans by body lice. B. quintana has adapted to both the human host and body louse vector niches, producing persistent infection with high titer bacterial loads in both the host (up to 105 colony-forming units [CFU]/ml) and vector (more than 108 CFU/ml). Using a novel custom microarray platform, we analyzed bacterial transcription at temperatures corresponding to the host (37°C) and vector (28°C), to probe for temperature-specific and growth phase-specific transcriptomes. We observed that transcription of 7% (93 genes) of the B. quintana genome is modified in response to change in growth phase, and that 5% (68 genes) of the genome is temperature-responsive. Among these transcriptional changes in response to temperature shift and growth phase was the induction of known B. quintana virulence genes and several previously unannotated genes. Hemin binding proteins, secretion systems, response regulators, and genes for invasion and cell attachment were prominent among the differentially-regulated B. quintana genes. This study represents the first analysis of global transcriptional responses by B. quintana. In addition, the in vivo experiments provide novel insight into the B. quintana transcriptional program within the body louse environment. These data and approaches will facilitate study of the adaptation mechanisms employed by Bartonella during the transition between human host and arthropod vector. PMID:23554923

Clinical value of patient-specific three-dimensional printing of congenital heart disease: Quantitative and qualitative assessments

PubMed Central

Lau, Ivan Wen Wen; Liu, Dongting; Xu, Lei; Fan, Zhanming

2018-01-01

Objective Current diagnostic assessment tools remain suboptimal in demonstrating complex morphology of congenital heart disease (CHD). This limitation has posed several challenges in preoperative planning, communication in medical practice, and medical education. This study aims to investigate the dimensional accuracy and the clinical value of 3D printed model of CHD in the above three areas. Methods Using cardiac computed tomography angiography (CCTA) data, a patient-specific 3D model of a 20-month-old boy with double outlet right ventricle was printed in Tango Plus material. Pearson correlation coefficient was used to evaluate correlation of the quantitative measurements taken at analogous anatomical locations between the CCTA images pre- and post-3D printing. Qualitative analysis was conducted by distributing surveys to six health professionals (two radiologists, two cardiologists and two cardiac surgeons) and three medical academics to assess the clinical value of the 3D printed model in these three areas. Results Excellent correlation (r = 0.99) was noted in the measurements between CCTA and 3D printed model, with a mean difference of 0.23 mm. Four out of six health professionals found the model to be useful in facilitating preoperative planning, while all of them thought that the model would be invaluable in enhancing patient-doctor communication. All three medical academics found the model to be helpful in teaching, and thought that the students will be able to learn the pathology quicker with better understanding. Conclusion The complex cardiac anatomy can be accurately replicated in flexible material using 3D printing technology. 3D printed heart models could serve as an excellent tool in facilitating preoperative planning, communication in medical practice, and medical education, although further studies with inclusion of more clinical cases are needed. PMID:29561912

Robot Faces that Follow Gaze Facilitate Attentional Engagement and Increase Their Likeability.

PubMed

Willemse, Cesco; Marchesi, Serena; Wykowska, Agnieszka

2018-01-01

Gaze behavior of humanoid robots is an efficient mechanism for cueing our spatial orienting, but less is known about the cognitive-affective consequences of robots responding to human directional cues. Here, we examined how the extent to which a humanoid robot (iCub) avatar directed its gaze to the same objects as our participants affected engagement with the robot, subsequent gaze-cueing, and subjective ratings of the robot's characteristic traits. In a gaze-contingent eyetracking task, participants were asked to indicate a preference for one of two objects with their gaze while an iCub avatar was presented between the object photographs. In one condition, the iCub then shifted its gaze toward the object chosen by a participant in 80% of the trials (joint condition) and in the other condition it looked at the opposite object 80% of the time (disjoint condition). Based on the literature in human-human social cognition, we took the speed with which the participants looked back at the robot as a measure of facilitated reorienting and robot-preference, and found these return saccade onset times to be quicker in the joint condition than in the disjoint condition. As indicated by results from a subsequent gaze-cueing tasks, the gaze-following behavior of the robot had little effect on how our participants responded to gaze cues. Nevertheless, subjective reports suggested that our participants preferred the iCub following participants' gaze to the one with a disjoint attention behavior, rated it as more human-like and as more likeable. Taken together, our findings show a preference for robots who follow our gaze. Importantly, such subtle differences in gaze behavior are sufficient to influence our perception of humanoid agents, which clearly provides hints about the design of behavioral characteristics of humanoid robots in more naturalistic settings.

Use of photogrammetry as a tool for accident investigation and reconstruction : a review of the literature and state of the practice.

DOT National Transportation Integrated Search

2007-01-01

One of the key goals of incident management is the rapid clearance of traffic incidents. Many transportation agencies focus their incident management on quicker and more reliable detection techniques. Another focus, however, should be the reduction i...

Emergency Medical Services (EMS) Public Information, Education and Relations (PIER): National Standard Curriculum (Administrator's Guide)

DOT National Transportation Integrated Search

1997-01-01

Imagine a transportation network linked by information and technology that makes travel quicker, safer and easier. Imagine being able to plot your route before you leave your home or office and know exactly how long it will take you - by car, subway ...

Savvy Consumers through Science

ERIC Educational Resources Information Center

Kahn, Sami

2005-01-01

Is Bounty the "quicker picker-upper?" Are expensive shampoos better? Are all antacids the same? The authors' fourth-grade students posed and answered these questions and many more during their recent "Consumer Product Testing" unit in which they designed experiments to assess these products' qualities and learned to question the advertising that…

Depletion of penicillin G residues in sows after intramuscular injection

USDA-ARS?s Scientific Manuscript database

In 2011, the Food Safety Inspection Service (FSIS) switched from using the Fast Antimicrobial Screen Test (FAST) for screening animal tissues for penicillin to using the Charm-Kidney Inhibition Swab test (KIS). The switch provided a quicker test and lower detection limits for penicillin when used o...

Brief Report: Signals Enhance the Suppressive Effects of Noncontingent Reinforcement

ERIC Educational Resources Information Center

Ringdahl, Joel E.; Call, Nathan A.; Christensen, Tory; Boelter, Eric W.

2010-01-01

The effects of noncontingent reinforcement (NCR) schedules on responding were assessed across two parameters: presence of signal and schedule density. Results indicated that signaled NCR schedules were correlated with greater overall reductions in responding and quicker reductions relative to NCR schedules without a signal. The clinical…

Chickenpox

MedlinePlus

... do get it after they've gotten the shot often get less severe cases, which means they get better quicker. What Happens When You Have Chickenpox? Chickenpox may start out seeming like a cold: You might have a runny or stuffy nose, sneezing, and a cough. But 1 to 2 days later, the rash begins, often ...

76 FR 52664 - Request for Information: Announcement of Carcinogen and Recommended Exposure Limit (REL) Policy...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-08-23

... in one or more experimental mammalian species as the result of any oral, respiratory or dermal... scientific evidence. In contrast, other organizations, such as the International Agency for Research on... need for more efficient and quicker means of classifying chemicals. Qualitative and semi- quantitative...

A Microgenetic Analysis of Strategic Variability in Gifted and Average-Ability Children

ERIC Educational Resources Information Center

Steiner, Hillary Hettinger

2006-01-01

Many researchers have described cognitive differences between gifted and average-performing children. Regarding strategy use, the gifted advantage is often associated with differences such as greater knowledge of strategies, quicker problem solving, and the ability to use strategies more appropriately. The current study used microgenetic methods…

Use of Google Earth to strengthen public health capacity and facilitate management of vector-borne diseases in resource-poor environments.

PubMed

Lozano-Fuentes, Saul; Elizondo-Quiroga, Darwin; Farfan-Ale, Jose Arturo; Loroño-Pino, Maria Alba; Garcia-Rejon, Julian; Gomez-Carro, Salvador; Lira-Zumbardo, Victor; Najera-Vazquez, Rosario; Fernandez-Salas, Ildefonso; Calderon-Martinez, Joaquin; Dominguez-Galera, Marco; Mis-Avila, Pedro; Morris, Natashia; Coleman, Michael; Moore, Chester G; Beaty, Barry J; Eisen, Lars

2008-09-01

Novel, inexpensive solutions are needed for improved management of vector-borne and other diseases in resource-poor environments. Emerging free software providing access to satellite imagery and simple editing tools (e.g. Google Earth) complement existing geographic information system (GIS) software and provide new opportunities for: (i) strengthening overall public health capacity through development of information for city infrastructures; and (ii) display of public health data directly on an image of the physical environment. We used freely accessible satellite imagery and a set of feature-making tools included in the software (allowing for production of polygons, lines and points) to generate information for city infrastructure and to display disease data in a dengue decision support system (DDSS) framework. Two cities in Mexico (Chetumal and Merida) were used to demonstrate that a basic representation of city infrastructure useful as a spatial backbone in a DDSS can be rapidly developed at minimal cost. Data layers generated included labelled polygons representing city blocks, lines representing streets, and points showing the locations of schools and health clinics. City blocks were colour-coded to show presence of dengue cases. The data layers were successfully imported in a format known as shapefile into a GIS software. The combination of Google Earth and free GIS software (e.g. HealthMapper, developed by WHO, and SIGEpi, developed by PAHO) has tremendous potential to strengthen overall public health capacity and facilitate decision support system approaches to prevention and control of vector-borne diseases in resource-poor environments.

Using a Novel Spatial Tool to Inform Invasive Species Early Detection and Rapid Response Efforts

NASA Astrophysics Data System (ADS)

Davidson, Alisha D.; Fusaro, Abigail J.; Kashian, Donna R.

2015-07-01

Management of invasive species has increasingly emphasized the importance of early detection and rapid response (EDRR) programs in limiting introductions, establishment, and impacts. These programs require an understanding of vector and species spatial dynamics to prioritize monitoring sites and efficiently allocate resources. Yet managers often lack the empirical data necessary to make these decisions. We developed an empirical mapping tool that can facilitate development of EDRR programs through identifying high-risk locations, particularly within the recreational boating vector. We demonstrated the utility of this tool in the Great Lakes watershed. We surveyed boaters to identify trips among water bodies and to quantify behaviors associated with high likelihood of species transfer (e.g., not removing organic materials from boat trailers) during that trip. We mapped water bodies with high-risk inbound and outbound boater movements using ArcGIS. We also tested for differences in high-risk behaviors based on demographic variables to understand risk differences among boater groups. Incorporation of boater behavior led to identification of additional high-risk water bodies compared to using the number of trips alone. Therefore, the number of trips itself may not fully reflect the likelihood of invasion. This tool can be broadly applied in other geographic contexts and with different taxa, and can be adjusted according to varying levels of information concerning the vector or species of interest. The methodology is straightforward and can be followed after a basic introduction to ArcGIS software. The visual nature of the mapping tool will facilitate site prioritization by managers and stakeholders from diverse backgrounds.

Direct Cloning of Yeast Genes from an Ordered Set of Lambda Clones in Saccharomyces Cerevisiae by Recombination in Vivo

PubMed Central

Erickson, J. R.; Johnston, M.

1993-01-01

We describe a technique that facilitates the isolation of yeast genes that are difficult to clone. This technique utilizes a plasmid vector that rescues lambda clones as yeast centromere plasmids. The source of these lambda clones is a set of clones whose location in the yeast genome has been determined by L. Riles et al. in 1993. The Esherichia coli-yeast shuttle plasmid carries URA3, ARS4 and CEN6, and contains DNA fragments from the lambda vector that flank the cloned yeast insert. When yeast is cotransformed with linearized plasmid and lambda clone DNA, Ura(+) transformants are obtained by a recombination event between the lambda clone and the plasmid vector that generates an autonomously replicating plasmid containing the cloned yeast DNA sequences. Genes whose genetic map positions are known can easily be identified and recovered in this plasmid by testing only those lambda clones that map to the relevant region of the yeast genome for their ability to complement the mutant phenotype. This technique facilitates the isolation of yeast genes that resist cloning either because (1) they are underrepresented in yeast genomic libraries amplified in E. coli, (2) they provide phenotypes that are too marginal to allow selection of the gene by genetic complementation or (3) they provide phenotypes that are laborious to score. We demonstrate the utility of this technique by isolating three genes, GAL83, SSN2 and MAK7, each of which presents one of these problems for cloning. PMID:8514124

Synthesis of an Uncharged Tetra-cyclopeptide Acting as a Transmembrane Carrier: Enhanced Cellular and Nuclear Uptake.

PubMed

Francisco Hilário, Flaviane; Traoré, Mohamed Dit Mady; Zwick, Vincent; Berry, Laurence; Simões-Pires, Claudia A; Cuendet, Muriel; Fantozzi, Nicolas; Pereira de Freitas, Rossimiriam; Maynadier, Marjorie; Wein, Sharon; Vial, Henri; Wong, Yung-Sing

2017-02-03

A small uncharged cyclopeptide scaffold inspired by a natural product and designed to undergo postfunctionalizations was used as a new transmembrane vector. A bioactive and fluorescent triazole aminocoumarin was bound to this carrier to facilitate its moving across cell and subcellular membranes, and this led to an increase in its cell toxicity.

78 FR 34554 - Establishment of Class E Airspace; Blue Mesa, CO

Federal Register 2010, 2011, 2012, 2013, 2014

2013-06-10

...This action establishes Class E airspace at Blue Mesa VHF Omni-Directional Radio Range/Distance Measuring Equipment (VOR/DME), Blue Mesa, CO, to facilitate vectoring of Instrument Flight Rules (IFR) aircraft under control of Denver and Albuquerque Air Route Traffic Control Centers (ARTCCs). This improves the safety and management of IFR operations within the National Airspace System.

Computational Approaches to Image Understanding.

DTIC Science & Technology

1981-10-01

represnting points, edges, surfaces, and volumes to facilitate display. The geometry or perspective and parailcl (or orthographic) projection has...of making the image forming process explicit. This in turn leads to a concern with geometry , such as the properties f the gradient, stereographic, and...dual spaces. Combining geometry and smoothness leads naturally to multi-variate vector analysis, and to differential geometry . For the most part, a

Facilitating protein solubility by use of peptide extensions

DOEpatents

Freimuth, Paul I; Zhang, Yian-Biao; Howitt, Jason

2013-09-17

Expression vectors for expression of a protein or polypeptide of interest as a fusion product composed of the protein or polypeptide of interest fused at one terminus to a solubility enhancing peptide extension are provided. Sequences encoding the peptide extensions are provided. The invention further comprises antibodies which bind specifically to one or more of the solubility enhancing peptide extensions.

Oak wilt: a potential threat to southern and western oak forests

Treesearch

A. Dan Wilson

2001-01-01

Oak wilt is recognized as one of the most destructive diseases to afflict oak species in the United States. The distribution and development of oak wilt in our eastern and midwestern oak forests has been closely linked to changes in forest stand composition, forest management practices, and pathogen dissemination facilitated by human and vector activity. The potential...

Automated astatination of biomolecules – a stepping stone towards multicenter clinical trials

PubMed Central

Aneheim, Emma; Albertsson, Per; Bäck, Tom; Jensen, Holger; Palm, Stig; Lindegren, Sture

2015-01-01

To facilitate multicentre clinical studies on targeted alpha therapy, it is necessary to develop an automated, on-site procedure for conjugating rare, short-lived, alpha-emitting radionuclides to biomolecules. Astatine-211 is one of the few alpha-emitting nuclides with appropriate chemical and physical properties for use in targeted therapies for cancer. Due to the very short range of the emitted α-particles, this therapy is particularly suited to treating occult, disseminated cancers. Astatine is not intrinsically tumour-specific; therefore, it requires an appropriate tumour-specific targeting vector, which can guide the radiation to the cancer cells. Consequently, an appropriate method is required for coupling the nuclide to the vector. To increase the availability of astatine-211 radiopharmaceuticals for targeted alpha therapy, their production should be automated. Here, we present a method that combines dry distillation of astatine-211 and a synthesis module for producing radiopharmaceuticals into a process platform. This platform will standardize production of astatinated radiopharmaceuticals, and hence, it will facilitate large clinical studies focused on this promising, but chemically challenging, alpha-emitting radionuclide. In this work, we describe the process platform, and we demonstrate the production of both astaine-211, for preclinical use, and astatine-211 labelled antibodies. PMID:26169786

Clustering, climate and dengue transmission.

PubMed

Junxiong, Pang; Yee-Sin, Leo

2015-06-01

Dengue is currently the most rapidly spreading vector-borne disease, with an increasing burden over recent decades. Currently, neither a licensed vaccine nor an effective anti-viral therapy is available, and treatment largely remains supportive. Current vector control strategies to prevent and reduce dengue transmission are neither efficient nor sustainable as long-term interventions. Increased globalization and climate change have been reported to influence dengue transmission. In this article, we reviewed the non-climatic and climatic risk factors which facilitate dengue transmission. Sustainable and effective interventions to reduce the increasing threat from dengue would require the integration of these risk factors into current and future prevention strategies, including dengue vaccination, as well as the continuous support and commitment from the political and environmental stakeholders.

A robust gene-stacking method utilizing yeast assembly for plant synthetic biology

DOE Office of Scientific and Technical Information (OSTI.GOV)

Shih, Patrick M.; Vuu, Khanh; Mansoori, Nasim

The advent and growth of synthetic biology has demonstrated its potential as a promising avenue of research to address many societal needs. But, plant synthetic biology efforts have been hampered by a dearth of DNA part libraries, versatile transformation vectors and efficient assembly strategies. We describe a versatile system (named jStack) utilizing yeast homologous recombination to efficiently assemble DNA into plant transformation vectors. We also demonstrate how this method can facilitate pathway engineering of molecules of pharmaceutical interest, production of potential biofuels and shuffling of disease-resistance traits between crop species. Our approach provides a powerful alternative to conventional strategies formore » stacking genes and traits to address many impending environmental and agricultural challenges.« less

A robust gene-stacking method utilizing yeast assembly for plant synthetic biology

DOE PAGES

Shih, Patrick M.; Vuu, Khanh; Mansoori, Nasim; ...

2016-10-26

The advent and growth of synthetic biology has demonstrated its potential as a promising avenue of research to address many societal needs. But, plant synthetic biology efforts have been hampered by a dearth of DNA part libraries, versatile transformation vectors and efficient assembly strategies. We describe a versatile system (named jStack) utilizing yeast homologous recombination to efficiently assemble DNA into plant transformation vectors. We also demonstrate how this method can facilitate pathway engineering of molecules of pharmaceutical interest, production of potential biofuels and shuffling of disease-resistance traits between crop species. Our approach provides a powerful alternative to conventional strategies formore » stacking genes and traits to address many impending environmental and agricultural challenges.« less

The developmental transcriptome of the mosquito Aedes aegypti, an invasive species and major arbovirus vector.

PubMed

Akbari, Omar S; Antoshechkin, Igor; Amrhein, Henry; Williams, Brian; Diloreto, Race; Sandler, Jeremy; Hay, Bruce A

2013-09-04

Mosquitoes are vectors of a number of important human and animal diseases. The development of novel vector control strategies requires a thorough understanding of mosquito biology. To facilitate this, we used RNA-seq to identify novel genes and provide the first high-resolution view of the transcriptome throughout development and in response to blood feeding in a mosquito vector of human disease, Aedes aegypti, the primary vector for Dengue and yellow fever. We characterized mRNA expression at 34 distinct time points throughout Aedes development, including adult somatic and germline tissues, by using polyA+ RNA-seq. We identify a total of 14,238 novel new transcribed regions corresponding to 12,597 new loci, as well as many novel transcript isoforms of previously annotated genes. Altogether these results increase the annotated fraction of the transcribed genome into long polyA+ RNAs by more than twofold. We also identified a number of patterns of shared gene expression, as well as genes and/or exons expressed sex-specifically or sex-differentially. Expression profiles of small RNAs in ovaries, early embryos, testes, and adult male and female somatic tissues also were determined, resulting in the identification of 38 new Aedes-specific miRNAs, and ~291,000 small RNA new transcribed regions, many of which are likely to be endogenous small-interfering RNAs and Piwi-interacting RNAs. Genes of potential interest for transgene-based vector control strategies also are highlighted. Our data have been incorporated into a user-friendly genome browser located at www.Aedes.caltech.edu, with relevant links to Vectorbase (www.vectorbase.org).

Improved Production Efficiency of Virus-Like Particles by the Baculovirus Expression Vector System

PubMed Central

Bárcena, Juan; Nuñez, Maria del Carmen; Martínez-Alonso, Diego; Dudognon, Benoit; Guijarro, Eva; Escribano, José M.

2015-01-01

Vaccines based on virus-like particles (VLPs) have proven effective in humans and animals. In this regard, the baculovirus expression vector system (BEVS) is one of the technologies of choice to generate such highly immunogenic vaccines. The extended use of these vaccines for human and animal populations is constrained because of high production costs, therefore a significant improvement in productivity is crucial to ensure their commercial viability. Here we describe the use of the previously described baculovirus expression cassette, called TB, to model the production of two VLP-forming vaccine antigens in insect cells. Capsid proteins from porcine circovirus type 2 (PCV2 Cap) and from the calicivirus that causes rabbit hemorrhagic disease (RHDV VP60) were expressed in insect cells using baculoviruses genetically engineered with the TB expression cassette. Productivity was compared to that obtained using standard counterpart vectors expressing the same proteins under the control of the polyhedrin promoter. Our results demonstrate that the use of the TB expression cassette increased the production yields of these vaccine antigens by around 300% with respect to the standard vectors. The recombinant proteins produced by TB-modified vectors were fully functional, forming VLPs identical in size and shape to those generated by the standard baculoviruses, as determined by electron microscopy analysis. The use of the TB expression cassette implies a simple modification of the baculovirus vectors that significantly improves the cost efficiency of VLP-based vaccine production, thereby facilitating the commercial viability and broad application of these vaccines for human and animal health. PMID:26458221

Diagnosis of nutrient imbalances with vector analysis in agroforestry systems.

PubMed

Isaac, Marney E; Kimaro, Anthony A

2011-01-01

Agricultural intensification has had unintended environmental consequences, including increased nutrient leaching and surface runoff and other agrarian-derived pollutants. Improved diagnosis of on-farm nutrient dynamics will have the advantage of increasing yields and will diminish financial and environmental costs. To achieve this, a management support system that allows for site-specific rapid evaluation of nutrient production imbalances and subsequent management prescriptions is needed for agroecological design. Vector diagnosis, a bivariate model to depict changes in yield and nutritional response simultaneously in a single graph, facilitates identification of nutritional status such as growth dilution, deficiency, sufficiency, luxury uptake, and toxicity. Quantitative data from cocoa agroforestry systems and pigeonpea intercropping trials in Ghana and Tanzania, respectively, were re-evaluated with vector analysis. Relative to monoculture, biomass increase in cocoa ( L.) under shade (35-80%) was accompanied by a 17 to 25% decline in P concentration, the most limiting nutrient on this site. Similarly, increasing biomass with declining P concentrations was noted for pigeonpea [ (L). Millsp.] in response to soil moisture availability under intercropping. Although vector analysis depicted nutrient responses, the current vector model does not consider non-nutrient resource effects on growth, such as ameliorated light and soil moisture, which were particularly active in these systems. We revisit and develop vector analysis into a framework for diagnosing nutrient and non-nutrient interactions in agroforestry systems. Such a diagnostic technique advances management decision-making by increasing nutrient precision and reducing environmental issues associated with agrarian-derived soil contamination. American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America.

Improved Production Efficiency of Virus-Like Particles by the Baculovirus Expression Vector System.

PubMed

López-Vidal, Javier; Gómez-Sebastián, Silvia; Bárcena, Juan; Nuñez, Maria del Carmen; Martínez-Alonso, Diego; Dudognon, Benoit; Guijarro, Eva; Escribano, José M

2015-01-01

Vaccines based on virus-like particles (VLPs) have proven effective in humans and animals. In this regard, the baculovirus expression vector system (BEVS) is one of the technologies of choice to generate such highly immunogenic vaccines. The extended use of these vaccines for human and animal populations is constrained because of high production costs, therefore a significant improvement in productivity is crucial to ensure their commercial viability. Here we describe the use of the previously described baculovirus expression cassette, called TB, to model the production of two VLP-forming vaccine antigens in insect cells. Capsid proteins from porcine circovirus type 2 (PCV2 Cap) and from the calicivirus that causes rabbit hemorrhagic disease (RHDV VP60) were expressed in insect cells using baculoviruses genetically engineered with the TB expression cassette. Productivity was compared to that obtained using standard counterpart vectors expressing the same proteins under the control of the polyhedrin promoter. Our results demonstrate that the use of the TB expression cassette increased the production yields of these vaccine antigens by around 300% with respect to the standard vectors. The recombinant proteins produced by TB-modified vectors were fully functional, forming VLPs identical in size and shape to those generated by the standard baculoviruses, as determined by electron microscopy analysis. The use of the TB expression cassette implies a simple modification of the baculovirus vectors that significantly improves the cost efficiency of VLP-based vaccine production, thereby facilitating the commercial viability and broad application of these vaccines for human and animal health.

Visualization of Microbiota in Tick Guts by Whole-mount In Situ Hybridization.

PubMed

Moss, Caitlin E; Robson, Andrew; Fikrig, Erol; Narasimhan, Sukanya

2018-06-01

Infectious diseases transmitted by arthropod vectors continue to pose a significant threat to human health worldwide. The pathogens causing these diseases, do not exist in isolation when they colonize the vector; rather, they likely engage in interactions with resident microorganisms in the gut lumen. The vector microbiota has been demonstrated to play an important role in pathogen transmission for several vector-borne diseases. Whether resident bacteria in the gut of the Ixodes scapularis tick, the vector of several human pathogens including Borrelia burgdorferi, influence tick transmission of pathogens is not determined. We require methods for characterizing the composition of the bacteria associated with the tick gut to facilitate a better understanding of potential interspecies interactions in the tick gut. Using whole-mount in situ hybridization to visualize RNA transcripts associated with particular bacterial species allows for the collection of qualitative data regarding the abundance and distribution of the microbiota in intact tissue. This technique can be used to examine changes in the gut microbiota milieu over the course of tick feeding and can also be applied to analyze expression of tick genes. Staining of whole tick guts yield information about the gross spatial distribution of target RNA in the tissue without the need for three-dimensional reconstruction and is less affected by environmental contamination, which often confounds the sequencing-based methods frequently used to study complex microbial communities. Overall, this technique is a valuable tool that can be used to better understand vector-pathogen-microbiota interactions and their role in disease transmission.

Diptera vectors of avian Haemosporidian parasites: untangling parasite life cycles and their taxonomy.

PubMed

Santiago-Alarcon, Diego; Palinauskas, Vaidas; Schaefer, Hinrich Martin

2012-11-01

Haemosporida is a large group of vector-borne intracellular parasites that infect amphibians, reptiles, birds, and mammals. This group includes the different malaria parasites (Plasmodium spp.) that infect humans around the world. Our knowledge on the full life cycle of these parasites is most complete for those parasites that infect humans and, to some extent, birds. However, our current knowledge on haemosporidian life cycles is characterized by a paucity of information concerning the vector species responsible for their transmission among vertebrates. Moreover, our taxonomic and systematic knowledge of haemosporidians is far from complete, in particular because of insufficient sampling in wild vertebrates and in tropical regions. Detailed experimental studies to identify avian haemosporidian vectors are uncommon, with only a few published during the last 25 years. As such, little knowledge has accumulated on haemosporidian life cycles during the last three decades, hindering progress in ecology, evolution, and systematic studies of these avian parasites. Nonetheless, recently developed molecular tools have facilitated advances in haemosporidian research. DNA can now be extracted from vectors' blood meals and the vertebrate host identified; if the blood meal is infected by haemosporidians, the parasite's genetic lineage can also be identified. While this molecular tool should help to identify putative vector species, detailed experimental studies on vector competence are still needed. Furthermore, molecular tools have helped to refine our knowledge on Haemosporida taxonomy and systematics. Herein we review studies conducted on Diptera vectors transmitting avian haemosporidians from the late 1800s to the present. We also review work on Haemosporida taxonomy and systematics since the first application of molecular techniques and provide recommendations and suggest future research directions. Because human encroachment on natural environments brings human populations into contact with novel parasite sources, we stress that the best way to avoid emergent and reemergent diseases is through a program encompassing ecological restoration, environmental education, and enhanced understanding of the value of ecosystem services. © 2012 The Authors. Biological Reviews © 2012 Cambridge Philosophical Society.

Enrichment of human hematopoietic stem/progenitor cells facilitates transduction for stem cell gene therapy.

PubMed

Baldwin, Kismet; Urbinati, Fabrizia; Romero, Zulema; Campo-Fernandez, Beatriz; Kaufman, Michael L; Cooper, Aaron R; Masiuk, Katelyn; Hollis, Roger P; Kohn, Donald B

2015-05-01

Autologous hematopoietic stem cell (HSC) gene therapy for sickle cell disease has the potential to treat this illness without the major immunological complications associated with allogeneic transplantation. However, transduction efficiency by β-globin lentiviral vectors using CD34-enriched cell populations is suboptimal and large vector production batches may be needed for clinical trials. Transducing a cell population more enriched for HSC could greatly reduce vector needs and, potentially, increase transduction efficiency. CD34(+) /CD38(-) cells, comprising ∼1%-3% of all CD34(+) cells, were isolated from healthy cord blood CD34(+) cells by fluorescence-activated cell sorting and transduced with a lentiviral vector expressing an antisickling form of beta-globin (CCL-β(AS3) -FB). Isolated CD34(+) /CD38(-) cells were able to generate progeny over an extended period of long-term culture (LTC) compared to the CD34(+) cells and required up to 40-fold less vector for transduction compared to bulk CD34(+) preparations containing an equivalent number of CD34(+) /CD38(-) cells. Transduction of isolated CD34(+) /CD38(-) cells was comparable to CD34(+) cells measured by quantitative PCR at day 14 with reduced vector needs, and average vector copy/cell remained higher over time for LTC initiated from CD34(+) /38(-) cells. Following in vitro erythroid differentiation, HBBAS3 mRNA expression was similar in cultures derived from CD34(+) /CD38(-) cells or unfractionated CD34(+) cells. In vivo studies showed equivalent engraftment of transduced CD34(+) /CD38(-) cells when transplanted in competition with 100-fold more CD34(+) /CD38(+) cells. This work provides initial evidence for the beneficial effects from isolating human CD34(+) /CD38(-) cells to use significantly less vector and potentially improve transduction for HSC gene therapy. © 2015 AlphaMed Press.

National malaria vector control policy: an analysis of the decision to scale-up larviciding in Nigeria.

PubMed

Tesfazghi, Kemi; Hill, Jenny; Jones, Caroline; Ranson, Hilary; Worrall, Eve

2016-02-01

New vector control tools are needed to combat insecticide resistance and reduce malaria transmission. The World Health Organization (WHO) endorses larviciding as a supplementary vector control intervention using larvicides recommended by the WHO Pesticides Evaluation Scheme (WHOPES). The decision to scale-up larviciding in Nigeria provided an opportunity to investigate the factors influencing policy adoption and assess the role that actors and evidence play in the policymaking process, in order to draw lessons that help accelerate the uptake of new methods for vector control. A retrospective policy analysis was carried out using in-depth interviews with national level policy stakeholders to establish normative national vector control policy or strategy decision-making processes and compare these with the process that led to the decision to scale-up larviciding. The interviews were transcribed, then coded and analyzed using NVivo10. Data were coded according to pre-defined themes from an analytical policy framework developed a priori. Stakeholders reported that the larviciding decision-making process deviated from the normative vector control decision-making process. National malaria policy is normally strongly influenced by WHO recommendations, but the potential of larviciding to contribute to national economic development objectives through larvicide production in Nigeria was cited as a key factor shaping the decision. The larviciding decision involved a restricted range of policy actors, and notably excluded actors that usually play advisory, consultative and evidence generation roles. Powerful actors limited the access of some actors to the policy processes and content. This may have limited the influence of scientific evidence in this policy decision. This study demonstrates that national vector control policy change can be facilitated by linking malaria control objectives to wider socioeconomic considerations and through engaging powerful policy champions to drive policy change and thereby accelerate access to new vector control tools. © The Author 2015. Published by Oxford University Press in association with The London School of Hygiene and Tropical Medicine.

An unsupervised machine learning model for discovering latent infectious diseases using social media data.

PubMed

Lim, Sunghoon; Tucker, Conrad S; Kumara, Soundar

2017-02-01

The authors of this work propose an unsupervised machine learning model that has the ability to identify real-world latent infectious diseases by mining social media data. In this study, a latent infectious disease is defined as a communicable disease that has not yet been formalized by national public health institutes and explicitly communicated to the general public. Most existing approaches to modeling infectious-disease-related knowledge discovery through social media networks are top-down approaches that are based on already known information, such as the names of diseases and their symptoms. In existing top-down approaches, necessary but unknown information, such as disease names and symptoms, is mostly unidentified in social media data until national public health institutes have formalized that disease. Most of the formalizing processes for latent infectious diseases are time consuming. Therefore, this study presents a bottom-up approach for latent infectious disease discovery in a given location without prior information, such as disease names and related symptoms. Social media messages with user and temporal information are extracted during the data preprocessing stage. An unsupervised sentiment analysis model is then presented. Users' expressions about symptoms, body parts, and pain locations are also identified from social media data. Then, symptom weighting vectors for each individual and time period are created, based on their sentiment and social media expressions. Finally, latent-infectious-disease-related information is retrieved from individuals' symptom weighting vectors. Twitter data from August 2012 to May 2013 are used to validate this study. Real electronic medical records for 104 individuals, who were diagnosed with influenza in the same period, are used to serve as ground truth validation. The results are promising, with the highest precision, recall, and F 1 score values of 0.773, 0.680, and 0.724, respectively. This work uses individuals' social media messages to identify latent infectious diseases, without prior information, quicker than when the disease(s) is formalized by national public health institutes. In particular, the unsupervised machine learning model using user, textual, and temporal information in social media data, along with sentiment analysis, identifies latent infectious diseases in a given location. Copyright © 2016 Elsevier Inc. All rights reserved.

Development of inducer-free expression plasmids based on IPTG-inducible promoters for Bacillus subtilis.

PubMed

Tran, Dinh Thi Minh; Phan, Trang Thi Phuong; Huynh, Thanh Kieu; Dang, Ngan Thi Kim; Huynh, Phuong Thi Kim; Nguyen, Tri Minh; Truong, Tuom Thi Tinh; Tran, Thuoc Linh; Schumann, Wolfgang; Nguyen, Hoang Duc

2017-07-25

Besides Escherichia coli, Bacillus subtilis is an important bacterial species for the production of recombinant proteins. Recombinant genes are inserted into shuttle expression vectors which replicate in both E. coli and in B. subtilis. The ligation products are first transformed into E. coli cells, analyzed for correct insertions, and the correct recombinant plasmids are then transformed into B. subtilis. A major problem using E. coli cells can be the strong basal level of expression of the recombinant protein which may interfere with the stability of the cells. To minimize this problem, we developed strong expression vectors being repressed in E. coli and inducer-free in B. subtilis. In general, induction of IPTG-inducible expression vectors is determined by the regulatory lacI gene encoding the LacI repressor in combination with the lacO operator on the promoter. To investigate the inducer-free properties of the vectors, we constructed inducer-free expression plasmids by removing the lacI gene and characterized their properties. First, we examined the ability to repress a reporter gene in E. coli, which is a prominent property facilitating the construction of the expression vectors carrying a target gene. The β-galactosidase (bgaB gene) basal levels expressed from Pgrac01-bgaB could be repressed at least twice in the E. coli cloning strain. Second, the inducer-free production of BgaB from four different plasmids with the Pgrac01 promoter in B. subtilis was investigated. As expected, BgaB expression levels of inducer-free constructs are at least 37 times higher than that of the inducible constructs in the absence of IPTG, and comparable to those in the presence of the inducer. Third, using efficient IPTG-inducible expression vectors containing the strong promoter Pgrac100, we could convert them into inducer-free expression plasmids. The BgaB production levels from the inducer-free plasmid in the absence of the inducer were at least 4.5 times higher than that of the inducible vector using the same promoter. Finally, we used gfp as a reporter gene in combination with the two promoters Pgrac01 and Pgrac100 to test the new vector types. The GFP expression levels could be repressed at least 1.5 times for the Pgrac01-gfp+ inducer-free construct in E. coli. The inducer-free constructs Pgrac01-gfp+ and Pgrac100-gfp+ allowed GFP expression at high levels from 23 × 10 4 to 32 × 10 4 RFU units and 9-13% of total intracellular proteins. We could reconfirm the two major advantages of the new inducer-free expression plasmids: (1) Strong repression of the target gene expression in the E. coli cloning strain, and (2) production of the target protein at high levels in B. subtilis in the absence of the inducer. We propose a general strategy to generate inducer-free expression vector by using IPTG-inducible vectors, and more specifically we developed inducer-free expression plasmids using IPTG-inducible promoters in the absence of the LacI repressor. These plasmids could be an excellent choice for high-level production of recombinant proteins in B. subtilis without the addition of inducer and at the same time maintaining a low basal level of the recombinant proteins in E. coli. The repression of the recombinant gene expression would facilitate cloning of genes that potentially inhibit the growth of E. coli cloning strains. The inducer-free expression plasmids will be extended versions of the current available IPTG-inducible expression vectors for B. subtilis, in which all these vectors use the same cognate promoters. These inducer-free and previously developed IPTG-inducible expression plasmids will be a useful cassette to study gene expression at a small scale up to a larger scale up for the production of recombinant proteins.

Effect of ionic strength on the interfacial viscoelasticity and stability of silk fibroin at the oil/water interface.

PubMed

Tang, Xiaoxiao; Qiao, Xiuying; Miller, Reinhard; Sun, Kang

2016-12-01

The amphiphilic character and surface activity endows silk fibroin with the ability to reside at fluid interfaces and effectively stabilize emulsions. However, the influence of relevant factors and their actual effect on the interfacial viscoelasticity and stability of silk fibroin at the oil/water interface has received less attention. In the present study, the effect of ionic strength on the interfacial viscoelasticity, emulsification effectiveness and stability of silk fibroin at the oil/water interface was investigated in detail. A higher ion concentration facilitates greater adsorption, stronger molecular interaction and faster structure reorganization of silk fibroin at the oil/water interface, thus causing quicker interfacial saturation adsorption, greater interfacial strength and lower interfacial structural fracture on large deformation. However, the presence of concentrated ions screens the charges in silk fibroin molecules and the zeta potential decreases as a result of electrostatic screening and ion-binding effects, which may result in emulsion droplet coalescence and a decrease in emulsion stability. The positively-charged ions significantly affect the interfacial elasticity and stability of silk fibroin layers at the oil/water interface as a result of the strong electrostatic interactions between counter-ions and the negatively-charged groups of silk fibroin. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

Use of an absorbable membrane to position biologically inductive materials in the periprosthetic space of cemented joints.

PubMed

DiResta, Gene R; Brown, Holly; Aiken, Sean; Doty, Steven; Schneider, Robert; Wright, Timothy; Healey, John H

2006-01-01

A device is presented that positions ultrahigh molecular weight polyethylene (UHMWPE) debris against periprosthetic bone surfaces. This can facilitate the study of aseptic loosening associated with cemented joint prostheses by speeding the appearance of this debris within the periprosthetic space. The device, composed of a 100 microm thick bioabsorbable membrane impregnated with 1.4 x 10(9) sub-micron particles of UHMWPE debris, is positioned on the endosteum of the bone prior to the insertion of the cemented orthopedic implant. An in vitro pullout study and an in vivo canine pilot study were performed to investigate its potential to accelerate "time to aseptic loosening" of cemented prosthetic joints. Pullout studies characterized the influence of the membrane on initial implant fixation. The tensile stresses (mean+/-std.dev.) required to withdraw a prosthesis cemented into canine femurs with and without the membrane were 1.15+/-0.3 and 1.54+/-0.01 MPa, respectively; these findings were not significantly different (p > 0.4). The in vivo pilot study, involving five dogs, was performed to evaluate the efficacy of the debris to accelerate loosening in a canine cemented hip arthroplasty. Aseptic loosening and lameness occurred within 12 months, quicker than the 30 months reported in a retrospective clinical review of canine hip arthroplasty.

Influence of tetracycline on tetracycline-resistant heterotrophs and tet genes in activated sludge process.

PubMed

Yu, Jie; Liu, Dongfang; Li, Kexun

2015-03-01

The concentrations of tetracycline-intermediate resistant, tetracycline-resistant heterotrophic bacteria, and total heterotrophic bacteria were examined to assess the influence of tetracycline on tetracycline-resistant heterotrophs by the R2A agar cultivation method in the tetracycline fortified activated sludge process and in the natural background. Results showed that the percentages of both tetracycline-intermediate resistant and tetracycline-resistant heterotrophic bacteria in total heterotrophic bacteria were significantly increased, after tetracycline was fed to activated sludge for a 3 months period under four different operating conditions, as compared with the background. In order to investigate the mechanism of activated sludge resistance to tetracycline, polymerase chain reaction experiments were carried out to analyze the existence and evolution of tet genes in the presence of tetracycline. Results revealed that only tet A and tet B genes out of the 11 target tet genes were observed in tetracycline treated activated sludge while no tet gene was detected in background. This indicated that tet A gene could accumulate in activated sludge with slower and continuous influent, while the accumulation of tet B gene could be attributed to shorter hydraulic retention time. Therefore, it was proposed in this study that tetracycline-resistant genes created by efflux pumps spread earlier and quicker to encode resistance to tetracycline, which facilitated the increase in tetracycline-resistance.

Plant pathogen nanodiagnostic techniques: forthcoming changes?

PubMed Central

Khiyami, Mohammad A.; Almoammar, Hassan; Awad, Yasser M.; Alghuthaymi, Mousa A.; Abd-Elsalam, Kamel A.

2014-01-01

Plant diseases are among the major factors limiting crop productivity. A first step towards managing a plant disease under greenhouse and field conditions is to correctly identify the pathogen. Current technologies, such as quantitative polymerase chain reaction (Q-PCR), require a relatively large amount of target tissue and rely on multiple assays to accurately identify distinct plant pathogens. The common disadvantage of the traditional diagnostic methods is that they are time consuming and lack high sensitivity. Consequently, developing low-cost methods to improve the accuracy and rapidity of plant pathogens diagnosis is needed. Nanotechnology, nano particles and quantum dots (QDs) have emerged as essential tools for fast detection of a particular biological marker with extreme accuracy. Biosensor, QDs, nanostructured platforms, nanoimaging and nanopore DNA sequencing tools have the potential to raise sensitivity, specificity and speed of the pathogen detection, facilitate high-throughput analysis, and to be used for high-quality monitoring and crop protection. Furthermore, nanodiagnostic kit equipment can easily and quickly detect potential serious plant pathogens, allowing experts to help farmers in the prevention of epidemic diseases. The current review deals with the application of nanotechnology for quicker, more cost-effective and precise diagnostic procedures of plant diseases. Such an accurate technology may help to design a proper integrated disease management system which may modify crop environments to adversely affect crop pathogens. PMID:26740775

Proposal for Using a Studio Format to Enhance Institutional Advancement

ERIC Educational Resources Information Center

Chance, Shannon

2008-01-01

Universities today need to become quicker on their toes. They must continually scan the environment and seize emerging opportunities--and institutional advancement must lead this effort. An unfortunate number of institutional advancement operations are ill equipped for the task at hand. Many suffer from high staff turnover and overly hierarchical…

Thirty Billion Quarters Can't Be Wrong--Or Can They?

ERIC Educational Resources Information Center

Needham, Nancy R.

1982-01-01

Social consequences likely to result from the popularity of video game arcades among teenagers are considered. Views of social theorists are summarized; social and financial problems created by the games are discussed, as are possible advantages, such as development of quicker reflexes. Attempts to regulate arcade hours are noted. (PP)

Mouse Click Plagiarism: Can Technology Help to Fight Back?

ERIC Educational Resources Information Center

Tulley Pitchford, Kay

2012-01-01

Many students arrive at university accustomed to adopting the internet as their primary source of information, but with no prior experience of referencing. This raises issues of the reliability and validity of digital sources, as well as bringing new opportunities for cheating. The internet has made plagiarism quicker and easier; a student simply…

PNNL Tests Fish Passage System

ScienceCinema

Colotelo, Alison

2018-01-16

Scientists from PNNL are testing a fish transportation system developed by Whooshh Innovations. The Whooshh system uses a flexible tube that works a bit like a vacuum, guiding fish over hydroelectric dams or other structures. Compared to methods used today, this system could save money while granting fish quicker, safer passage through dams and hatcheries.

Expanding the Acculturation Gap-Distress Model: An Integrative Review of Research

ERIC Educational Resources Information Center

Telzer, Eva H.

2010-01-01

The acculturation gap-distress model purports that immigrant children acculturate to their new culture at a quicker pace than their parents, leading to family conflict and youth maladjustment. This article reviews literature on the acculturation gap-distress model, showing that acculturation gaps function in unique ways depending on many social…

PNNL Tests Fish Passage System

DOE Office of Scientific and Technical Information (OSTI.GOV)

Colotelo, Alison

2015-03-13

Scientists from PNNL are testing a fish transportation system developed by Whooshh Innovations. The Whooshh system uses a flexible tube that works a bit like a vacuum, guiding fish over hydroelectric dams or other structures. Compared to methods used today, this system could save money while granting fish quicker, safer passage through dams and hatcheries.

Spatial Processes in Linear Ordering

ERIC Educational Resources Information Center

von Hecker, Ulrich; Klauer, Karl Christoph; Wolf, Lukas; Fazilat-Pour, Masoud

2016-01-01

Memory performance in linear order reasoning tasks (A > B, B > C, C > D, etc.) shows quicker, and more accurate responses to queries on wider (AD) than narrower (AB) pairs on a hypothetical linear mental model (A -- B -- C -- D). While indicative of an analogue representation, research so far did not provide positive evidence for spatial…

Combined ultraviolet and water exposure as a preconditioning method in laboratory fungal durability testing

Treesearch

Rebecca E. Ibach; Craig M. Clemons; Nicole M. Stark

2003-01-01

During outdoor exposure, woodfiber-plastic composites (WPC) are subject to biological, moisture, and ultraviolet (UV) degradation. The purpose of laboratory evaluations is to simulate outdoor conditions and accelerate the testing for quicker results. Traditionally, biological, moisture, and W laboratory tests are done separately, and only combined in outdoor field...

Taming the Saxophone

ERIC Educational Resources Information Center

Murphy, Sean

2013-01-01

The saxophone section of a wind ensemble can easily be one of the most frustrating to work with when it comes to producing a clear, characteristic tone. Sometimes, the road to an improved sound can be a long path of daily diligence and practice; however, there are many quicker solutions that will drastically improve a student's tone. This article…

Hyperspectral remote sensing of postfire soil properties

Treesearch

Sarah A. Lewis; Peter R. Robichaud; William J. Elliot; Bruce E. Frazier; Joan Q. Wu

2004-01-01

Forest fires may induce changes in soil organic properties that often lead to water repellent conditions within the soil profile that decrease soil infiltration capacity. The remote detection of water repellent soils after forest fires would lead to quicker and more accurate assessment of erosion potential. An airborne hyperspectral image was acquired over the Hayman...

Bilingual Welsh-English Children's Acquisition of Vocabulary and Reading: Implications for Bilingual Education

ERIC Educational Resources Information Center

Rhys, Mirain; Thomas, Enlli Môn

2013-01-01

Previous studies have highlighted early differences in bilinguals' rate of language acquisition in comparison with monolinguals. However, these differences seem to disappear with increasing age and exposure to the language, and do so quicker in dominant community languages than in minority status languages. This study aimed to replicate these…

Busting the New Breed of Plagiarist.

ERIC Educational Resources Information Center

Bugeja, Michael

2000-01-01

The new breed of plagiarists knows that stealing from the World Wide Web is quicker than stealing from the library at universities that typically provide online services. The new plagiarists have been weaned on chat rooms, guest books, news groups, mailing lists, MOOs, and MUDs--myriad online ways to procrastinate when final papers are due. The…

76 FR 71346 - Public Meeting and Request for Information: Carcinogen and Recommended Exposure Limit (REL...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-11-17

... experimental mammalian species as the result of any oral, respiratory or dermal exposure, or any other exposure... contrast, other organizations, such as the International Agency for Research on Cancer (IARC) and the... and quicker means of classifying chemicals. Qualitative and semi- quantitative approaches such as...

Block versus Random Amphiphilic Glycopolymer Nanopaticles as Glucose-Responsive Vehicles.

PubMed

Guo, Qianqian; Zhang, Tianqi; An, Jinxia; Wu, Zhongming; Zhao, Yu; Dai, Xiaomei; Zhang, Xinge; Li, Chaoxing

2015-10-12

To explore the effect of polymer structure on their self-assembled aggregates and their unique characteristics, this study was devoted to developing a series of amphiphilic block and random phenylboronic acid-based glycopolymers by RAFT polymerization. The amphiphilic glycopolymers were successfully self-assembled into spherically shaped nanoparticles with narrow size distribution in aqueous solution. For block and random copolymers with similar monomer compositions, block copolymer nanoparticles exhibited a more regular transmittance change with the increasing glucose level, while a more evident variation of size and quicker decreasing tendency in I/I0 behavior in different glucose media were observed for random copolymer nanoparticles. Cell viability of all the polymer nanoparticles investigated by MTT assay was higher than 80%, indicating that both block and random copolymers had good cytocompatibility. Insulin could be encapsulated into both nanoparticles, and insulin release rate for random glycopolymer was slightly quicker than that for the block ones. We speculate that different chain conformations between block and random glycopolymers play an important role in self-assembled nanoaggregates and underlying glucose-sensitive behavior.

Improving protein-protein interactions prediction accuracy using protein evolutionary information and relevance vector machine model.

PubMed

An, Ji-Yong; Meng, Fan-Rong; You, Zhu-Hong; Chen, Xing; Yan, Gui-Ying; Hu, Ji-Pu

2016-10-01

Predicting protein-protein interactions (PPIs) is a challenging task and essential to construct the protein interaction networks, which is important for facilitating our understanding of the mechanisms of biological systems. Although a number of high-throughput technologies have been proposed to predict PPIs, there are unavoidable shortcomings, including high cost, time intensity, and inherently high false positive rates. For these reasons, many computational methods have been proposed for predicting PPIs. However, the problem is still far from being solved. In this article, we propose a novel computational method called RVM-BiGP that combines the relevance vector machine (RVM) model and Bi-gram Probabilities (BiGP) for PPIs detection from protein sequences. The major improvement includes (1) Protein sequences are represented using the Bi-gram probabilities (BiGP) feature representation on a Position Specific Scoring Matrix (PSSM), in which the protein evolutionary information is contained; (2) For reducing the influence of noise, the Principal Component Analysis (PCA) method is used to reduce the dimension of BiGP vector; (3) The powerful and robust Relevance Vector Machine (RVM) algorithm is used for classification. Five-fold cross-validation experiments executed on yeast and Helicobacter pylori datasets, which achieved very high accuracies of 94.57 and 90.57%, respectively. Experimental results are significantly better than previous methods. To further evaluate the proposed method, we compare it with the state-of-the-art support vector machine (SVM) classifier on the yeast dataset. The experimental results demonstrate that our RVM-BiGP method is significantly better than the SVM-based method. In addition, we achieved 97.15% accuracy on imbalance yeast dataset, which is higher than that of balance yeast dataset. The promising experimental results show the efficiency and robust of the proposed method, which can be an automatic decision support tool for future proteomics research. For facilitating extensive studies for future proteomics research, we developed a freely available web server called RVM-BiGP-PPIs in Hypertext Preprocessor (PHP) for predicting PPIs. The web server including source code and the datasets are available at http://219.219.62.123:8888/BiGP/. © 2016 The Authors Protein Science published by Wiley Periodicals, Inc. on behalf of The Protein Society.

Bioinformatic prediction of G protein-coupled receptor encoding sequences from the transcriptome of the foreleg, including the Haller’s organ, of the cattle tick, Rhipicephalus australis

USDA-ARS?s Scientific Manuscript database

The cattle tick of Australia, Rhipicephalus australis, is a vector for microbial parasites that cause serious bovine diseases. The Haller's organ, located in the tick's forelegs, is crucial for host detection and mating. To facilitate the development of new technologies for better control of this ag...

Microsatellite characterization of subspecies and their hybrids in Culex pipiens complex mosquitoes along a north-south transect in the central United States of America

USDA-ARS?s Scientific Manuscript database

Mosquitoes in the Culex pipiens complex, Cx. p. pipiens L. (Cpp) and Cx. p. quinquefasciatus Say (Cpq), are morphologically similar and important vectors of the West Nile and St. Louis Encephalitis viruses in the US. They hybridize when found in sympatry, which could facilitate the transfer of adva...

Institutional Interpretations of the Relationship between Sport-Related Disciplines and Their Reference Disciplines: The Case of Sociology of Sport

ERIC Educational Resources Information Center

Ciomaga, Bogdan

2014-01-01

The sociology of sport literature is now sufficiently broad to allow a general analysis of research patterns in this field. To facilitate the identification of these vectors of expansion, sources referenced in articles published in "Sociology of Sport Journal and Journal of Sport and Social Issues" between 2003 and 2011 are examined in…

78 FR 78794 - Proposed Establishment of Class E Airspace; Flagstaff, AZ

Federal Register 2010, 2011, 2012, 2013, 2014

2013-12-27

...This action proposes to establish Class E airspace at the Flagstaff VHF Omni-Directional Radio Range/Distance Measuring Equipment (VOR/DME) navigation aid, Flagstaff, AZ, to facilitate vectoring of Instrument Flight Rules (IFR) aircraft under control of Albuquerque Air Route Traffic Control Center (ARTCC). The FAA is proposing this action to enhance the safety and management of aircraft operations within the National Airspace System.

[Vector transmitted diseases and climate changes in Europe].

PubMed

Rossati, Antonella; Bargiacchi, Olivia; Kroumova, Vesselina; Garavelli, Pietro Luigi

2014-09-01

The increase in temperatures recorded since the mid-nineteenth century is unprecedented in the history of mankind. The consequences of climate changes are numerous and can affect human health through direct (extreme events, natural disasters) or indirect (alteration of the ecosystem) mechanisms. Climate changes have repercussions on ecosystems, agriculture, social conditions, migration, conflicts and the transmission mode of infectious diseases. Vector-borne diseases are infections transmitted by the bite of infected arthropods such as mosquitoes, ticks, triatomines, sand flies and flies. Epidemiological cornerstones of vector-borne diseases are: the ecology and behaviour of the host, the ecology and behaviour of the vector, and the population's degree of immunity. Mosquito vectors related to human diseases mainly belong to the genus Culex, Aedes and Mansonia. Climate changes in Europe have increased the spread of new vectors, such as Aedes albopictus, and in some situations have made it possible to sustain the autochthonous transmission of some diseases (outbreak of Chukungunya virus in northern Italy in 2007, cases of dengue in the South of France and in Croatia). Despite the eradication of malaria from Europe, anopheline carriers are still present, and they may allow the transmission of the disease if the climatic conditions favour the development of the vectors and their contacts with plasmodium carriers. The tick Ixodes ricinus is a vector whose expansion has been documented both in latitude and in altitude in relation to the temperature increase; at the same time the related main viral and bacterial infections have increased. In northern Italy and Germany, the appearance of Leishmaniasis has been associated to climatic conditions that favour the development of the vector Phlebotomus papatasi and the maturation of the parasite within the vector, although the increase of cases of visceral leishmaniasis is also related to host immune factors, particularly immunodepression caused by the human immunodeficiency virus (HIV). Despite the importance of global warming in facilitating the transmission of certain infectious diseases, due consideration must be taken of the role played by other variables, such as the increase in international travel, migration and trade, with the risk of importing parasites and vectors with the goods. In addition, the control of certain infections was possible in the past through improvements in socio-economic conditions of affected populations. However, the reduction in resources allocated to health care has recently led to the re-emergence of diseases that were considered eradicated.

Utilization of Virus ϕCh1 Elements To Establish a Shuttle Vector System for Halo(alkali)philic Archaea via Transformation of Natrialba magadii

PubMed Central

Mayrhofer-Iro, M.; Ladurner, A.; Meissner, C.; Derntl, C.; Reiter, M.; Haider, F.; Dimmel, K.; Rössler, N.; Klein, R.; Baranyi, U.; Scholz, H.

2013-01-01

In the study described here, we successfully developed a transformation system for halo(alkali)philic members of the Archaea. This transformation system comprises a series of Natrialba magadii/Escherichia coli shuttle vectors based on a modified method to transform halophilic members of the Archaea and genomic elements of the N. magadii virus ϕCh1. The shuttle vector pRo-5, based on the repH-containing region of ϕCh1, stably replicated in E. coli and N. magadii and in several halophilic and haloalkaliphilic members of the Archaea not transformable so far. The ϕCh1 operon ORF53/ORF54 (repH) was essential for pRo-5 replication and was thus identified as the minimal replication origin. The plasmid allowed homologous and heterologous gene expression, as exemplified by the expression of ϕCh1 ORF3452, which encodes a structural protein, and the reporter gene bgaH of Haloferax lucentense in N. magadii. The new transformation/vector system will facilitate genetic studies within N. magadii and other haloalkaliphilic archaea and will allow the detailed characterization of the gene functions of N. magadii virus ϕCh1 in their extreme environments. PMID:23416999

UDE-based control of variable-speed wind turbine systems

NASA Astrophysics Data System (ADS)

Ren, Beibei; Wang, Yeqin; Zhong, Qing-Chang

2017-01-01

In this paper, the control of a PMSG (permanent magnet synchronous generator)-based variable-speed wind turbine system with a back-to-back converter is considered. The uncertainty and disturbance estimator (UDE)-based control approach is applied to the regulation of the DC-link voltage and the control of the RSC (rotor-side converter) and the GSC (grid-side converter). For the rotor-side controller, the UDE-based vector control is developed for the RSC with PMSG control to facilitate the application of the MPPT (maximum power point tracking) algorithm for the maximum wind energy capture. For the grid-side controller, the UDE-based vector control is developed to control the GSC with the power reference generated by a UDE-based DC-link voltage controller. Compared with the conventional vector control, the UDE-based vector control can achieve reliable current decoupling control with fast response. Moreover, the UDE-based DC-link voltage regulation can achieve stable DC-link voltage under model uncertainties and external disturbances, e.g. wind speed variations. The effectiveness of the proposed UDE-based control approach is demonstrated through extensive simulation studies in the presence of coupled dynamics, model uncertainties and external disturbances under varying wind speeds. The UDE-based control is able to generate more energy, e.g. by 5% for the wind profile tested.

Influence of molecular weight upon mannosylated bio-synthetic hybrids for targeted antigen presenting cell gene delivery.

PubMed

Jones, Charles H; Gollakota, Akhila; Chen, Mingfu; Chung, Tai-Chun; Ravikrishnan, Anitha; Zhang, Guojian; Pfeifer, Blaine A

2015-07-01

Given the rise of antibiotic resistant microbes, genetic vaccination is a promising prophylactic strategy that enables rapid design and manufacture. Facilitating this process is the choice of vector, which is often situationally-specific and limited in engineering capacity. Furthermore, these shortcomings are usually tied to an incomplete understanding of the structure-function relationships driving vector-mediated gene delivery. Building upon our initial report of a hybrid bacterial-biomaterial gene delivery vector, a comprehensive structure-function assessment was completed using a class of mannosylated poly(beta-amino esters). Through a top-down screening methodology, an ideal polymer was selected on the basis of gene delivery efficacy and then used for the synthesis of a stratified molecular weight polymer library. By eliminating contributions of polymer chemical background, we were able to complete an in-depth assessment of gene delivery as a function of (1) polymer molecular weight, (2) relative mannose content, (3) polymer-membrane biophysical properties, (4) APC uptake specificity, and (5) serum inhibition. In summary, the flexibility and potential of the hybrid design featured in this work highlights the ability to systematically probe vector-associated properties for the development of translational gene delivery candidates. Copyright © 2015 Elsevier Ltd. All rights reserved.

Weighted K-means support vector machine for cancer prediction.

PubMed

Kim, SungHwan

2016-01-01

To date, the support vector machine (SVM) has been widely applied to diverse bio-medical fields to address disease subtype identification and pathogenicity of genetic variants. In this paper, I propose the weighted K-means support vector machine (wKM-SVM) and weighted support vector machine (wSVM), for which I allow the SVM to impose weights to the loss term. Besides, I demonstrate the numerical relations between the objective function of the SVM and weights. Motivated by general ensemble techniques, which are known to improve accuracy, I directly adopt the boosting algorithm to the newly proposed weighted KM-SVM (and wSVM). For predictive performance, a range of simulation studies demonstrate that the weighted KM-SVM (and wSVM) with boosting outperforms the standard KM-SVM (and SVM) including but not limited to many popular classification rules. I applied the proposed methods to simulated data and two large-scale real applications in the TCGA pan-cancer methylation data of breast and kidney cancer. In conclusion, the weighted KM-SVM (and wSVM) increases accuracy of the classification model, and will facilitate disease diagnosis and clinical treatment decisions to benefit patients. A software package (wSVM) is publicly available at the R-project webpage (https://www.r-project.org).

Gateway Vectors for Efficient Artificial Gene Assembly In Vitro and Expression in Yeast Saccharomyces cerevisiae

PubMed Central

Giuraniuc, Claudiu V.; MacPherson, Murray; Saka, Yasushi

2013-01-01

Construction of synthetic genetic networks requires the assembly of DNA fragments encoding functional biological parts in a defined order. Yet this may become a time-consuming procedure. To address this technical bottleneck, we have created a series of Gateway shuttle vectors and an integration vector, which facilitate the assembly of artificial genes and their expression in the budding yeast Saccharomyces cerevisiae. Our method enables the rapid construction of an artificial gene from a promoter and an open reading frame (ORF) cassette by one-step recombination reaction in vitro. Furthermore, the plasmid thus created can readily be introduced into yeast cells to test the assembled gene’s functionality. As flexible regulatory components of a synthetic genetic network, we also created new versions of the tetracycline-regulated transactivators tTA and rtTA by fusing them to the auxin-inducible degron (AID). Using our gene assembly approach, we made yeast expression vectors of these engineered transactivators, AIDtTA and AIDrtTA and then tested their functions in yeast. We showed that these factors can be regulated by doxycycline and degraded rapidly after addition of auxin to the medium. Taken together, the method for combinatorial gene assembly described here is versatile and would be a valuable tool for yeast synthetic biology. PMID:23675537

Bottlenecks in domestic animal populations can facilitate the emergence of Trypanosoma cruzi, the aetiological agent of Chagas disease

PubMed Central

Levy, Michael Z.; Tustin, Aaron; Castillo-Neyra, Ricardo; Mabud, Tarub S.; Levy, Katelyn; Barbu, Corentin M.; Quispe-Machaca, Victor R.; Ancca-Juarez, Jenny; Borrini-Mayori, Katty; Naquira-Velarde, Cesar; Ostfeld, Richard S.

2015-01-01

Faeces-mediated transmission of Trypanosoma cruzi (the aetiological agent of Chagas disease) by triatomine insects is extremely inefficient. Still, the parasite emerges frequently, and has infected millions of people and domestic animals. We synthesize here the results of field and laboratory studies of T. cruzi transmission conducted in and around Arequipa, Peru. We document the repeated occurrence of large colonies of triatomine bugs (more than 1000) with very high infection prevalence (more than 85%). By inoculating guinea pigs, an important reservoir of T. cruzi in Peru, and feeding triatomine bugs on them weekly, we demonstrate that, while most animals quickly control parasitaemia, a subset of animals remains highly infectious to vectors for many months. However, we argue that the presence of these persistently infectious hosts is insufficient to explain the observed prevalence of T. cruzi in vector colonies. We posit that seasonal rains, leading to a fluctuation in the price of guinea pig food (alfalfa), leading to annual guinea pig roasts, leading to a concentration of vectors on a small subpopulation of animals maintained for reproduction, can propel T. cruzi through vector colonies and create a considerable force of infection for a pathogen whose transmission might otherwise fizzle out. PMID:26085582

Globally invasive, withdrawing at home: Aedes albopictus and Aedes japonicus facing the rise of Aedes flavopictus

NASA Astrophysics Data System (ADS)

Chaves, Luis Fernando

2016-11-01

It has been suggested that climate change may have facilitated the global expansion of invasive disease vectors, since several species have expanded their range as temperatures have warmed. Here, we present results from observations on two major global invasive mosquito vectors (Diptera: Culicidae), Aedes albopictus (Skuse) and Aedes japonicus (Theobald), across the altitudinal range of Mt. Konpira, Nagasaki, Japan, a location within their native range, where Aedes flavopictus Yamada, formerly a rare species, has now become dominant. Spatial abundance patterns of the three species suggest that temperature is an important factor influencing their adult distribution across the altitudinal range of Mt. Konpira. Temporal abundance patterns, by contrast, were associated with rainfall and showed signals of density-dependent regulation in the three species. The spatial and temporal analysis of abundance patterns showed that Ae. flavopictus and Ae. albopictus were negatively associated, even when accounting for differential impacts of weather and other environmental factors in their co-occurrence patterns. Our results highlight a contingency in the expansion of invasive vectors, the potential emergence of changes in their interactions with species in their native communities, and raise the question of whether these changes might be useful to predict the emergence of future invasive vectors.

Amiloride-enhanced gene transfection of octa-arginine functionalized calcium phosphate nanoparticles.

PubMed

Vanegas Sáenz, Juan Ramón; Tenkumo, Taichi; Kamano, Yuya; Egusa, Hiroshi; Sasaki, Keiichi

2017-01-01

Nanoparticles represent promising gene delivery systems in biomedicine to facilitate prolonged gene expression with low toxicity compared to viral vectors. Specifically, nanoparticles of calcium phosphate (nCaP), the main inorganic component of human bone, exhibit high biocompatibility and good biodegradability and have been reported to have high affinity for protein or DNA, having thus been used as gene transfer vectors. On the other hand, Octa-arginine (R8), which has a high permeability to cell membrane, has been reported to improve intracellular delivery systems. Here, we present an optimized method for nCaP-mediated gene delivery using an octa-arginine (R8)-functionalized nCaP vector containing a marker or functional gene construct. nCaP particle size was between 220-580 nm in diameter and all R8-functionalized nCaPs carried a positive charge. R8 concentration significantly improved nCaP transfection efficiency with high cell compatibility in human mesenchymal stem cells (hMSC) and human osteoblasts (hOB) in particular, suggesting nCaPs as a good option for non-viral vector gene delivery. Furthermore, pre-treatment with different endocytosis inhibitors identified that the endocytic pathway differed among cell lines and functionalized nanoparticles, with amiloride increasing transfection efficiency of R8-functionalized nCaPs in hMSC and hOB.

Bottlenecks in domestic animal populations can facilitate the emergence of Trypanosoma cruzi, the aetiological agent of Chagas disease.

PubMed

Levy, Michael Z; Tustin, Aaron; Castillo-Neyra, Ricardo; Mabud, Tarub S; Levy, Katelyn; Barbu, Corentin M; Quispe-Machaca, Victor R; Ancca-Juarez, Jenny; Borrini-Mayori, Katty; Naquira-Velarde, Cesar; Ostfeld, Richard S

2015-07-07

Faeces-mediated transmission of Trypanosoma cruzi (the aetiological agent of Chagas disease) by triatomine insects is extremely inefficient. Still, the parasite emerges frequently, and has infected millions of people and domestic animals. We synthesize here the results of field and laboratory studies of T. cruzi transmission conducted in and around Arequipa, Peru. We document the repeated occurrence of large colonies of triatomine bugs (more than 1000) with very high infection prevalence (more than 85%). By inoculating guinea pigs, an important reservoir of T. cruzi in Peru, and feeding triatomine bugs on them weekly, we demonstrate that, while most animals quickly control parasitaemia, a subset of animals remains highly infectious to vectors for many months. However, we argue that the presence of these persistently infectious hosts is insufficient to explain the observed prevalence of T. cruzi in vector colonies. We posit that seasonal rains, leading to a fluctuation in the price of guinea pig food (alfalfa), leading to annual guinea pig roasts, leading to a concentration of vectors on a small subpopulation of animals maintained for reproduction, can propel T. cruzi through vector colonies and create a considerable force of infection for a pathogen whose transmission might otherwise fizzle out. © 2015 The Author(s) Published by the Royal Society. All rights reserved.

Microneedle-mediated immunization of an adenovirus-based malaria vaccine enhances antigen-specific antibody immunity and reduces anti-vector responses compared to the intradermal route.

PubMed

Carey, John B; Vrdoljak, Anto; O'Mahony, Conor; Hill, Adrian V S; Draper, Simon J; Moore, Anne C

2014-08-21

Substantial effort has been placed in developing efficacious recombinant attenuated adenovirus-based vaccines. However induction of immunity to the vector is a significant obstacle to its repeated use. Here we demonstrate that skin-based delivery of an adenovirus-based malaria vaccine, HAdV5-PyMSP1₄₂, to mice using silicon microneedles induces equivalent or enhanced antibody responses to the encoded antigen, however it results in decreased anti-vector responses, compared to intradermal delivery. Microneedle-mediated vaccine priming and resultant induction of low anti-vector antibody titres permitted repeated use of the same adenovirus vaccine vector. This resulted in significantly increased antigen-specific antibody responses in these mice compared to ID-treated mice. Boosting with a heterologous vaccine; MVA-PyMSP1₄₂ also resulted in significantly greater antibody responses in mice primed with HAdV5-PyMSP1₄₂ using MN compared to the ID route. The highest protection against blood-stage malaria challenge was observed when a heterologous route of immunization (MN/ID) was used. Therefore, microneedle-mediated immunization has potential to both overcome some of the logistic obstacles surrounding needle-and-syringe-based immunization as well as to facilitate the repeated use of the same adenovirus vaccine thereby potentially reducing manufacturing costs of multiple vaccines. This could have important benefits in the clinical ease of use of adenovirus-based immunization strategies.

Legal aspects of public health: difficulties in controlling vector-borne and zoonotic diseases in Brazil.

PubMed

Mendes, Marcílio S; de Moraes, Josué

2014-11-01

In recent years, vector-borne and zoonotic diseases have become a major challenge for public health. Dengue fever and leptospirosis are the most important communicable diseases in Brazil based on their prevalence and the healthy life years lost from disability. The primary strategy for preventing human exposure to these diseases is effective insect and rodent control in and around the home. However, health authorities have difficulties in controlling vector-borne and zoonotic diseases because residents often refuse access to their homes. This study discusses aspects related to the activities performed by Brazilian health authorities to combat vector-borne and zoonotic diseases, particularly difficulties in relation to the legal aspect, which often impede the quick and effective actions of these professionals. How might it be possible to reconcile the need to preserve public health and the rule on the inviolability of the home, especially in the case of abandoned properties or illegal residents and the refusal of residents to allow the health authority access? Do residents have the right to hinder the performance of health workers even in the face of a significant and visible focus of disease transmission? This paper argues that a comprehensive legal plan aimed at the control of invasive vector-borne and zoonotic diseases including synanthropic animals of public health importance should be considered. In addition, this paper aims to bridge the gap between lawyers and public health professionals and to facilitate communication between them. Copyright © 2014 Elsevier B.V. All rights reserved.

Adapter-directed display: a modular design for shuttling display on phage surfaces.

PubMed

Wang, Kevin Caili; Wang, Xinwei; Zhong, Pingyu; Luo, Peter Peizhi

2010-02-05

A novel adapter-directed phage display system was developed with modular features. In this system, the target protein is expressed as a fusion protein consisting of adapter GR1 from the phagemid vector, while the recombinant phage coat protein is expressed as a fusion protein consisting of adapter GR2 in the helper phage vector. Surface display of the target protein is accomplished through specific heterodimerization of GR1 and GR2 adapters, followed by incorporation of the heterodimers into phage particles. A series of engineered helper phages were constructed to facilitate both display valency and formats, based on various phage coat proteins. As the target protein is independent of a specific phage coat protein, this modular system allows the target protein to be displayed on any given phage coat protein and allows various display formats from the same vector without the need for reengineering. Here, we demonstrate the shuttling display of a single-chain Fv antibody on phage surfaces between multivalent and monovalent formats, as well as the shuttling display of an antigen-binding fragment molecule on phage coat proteins pIII, pVII, and pVIII using the same phagemid vectors combined with different helper phage vectors. This adapter-directed display concept has been applied to eukaryotic yeast surface display and to a novel cross-species display that can shuttle between prokaryotic phage and eukaryotic yeast systems. Copyright 2009 Elsevier Ltd. All rights reserved.

First record of Stegomyia albopicta in Turkey determined by active ovitrap surveillance and DNA barcoding.

PubMed

Oter, Kerem; Gunay, Filiz; Tuzer, Erkut; Linton, Yvonne-Marie; Bellini, Romeo; Alten, Bulent

2013-10-01

Despite its confirmed establishment in neighboring Greece and Bulgaria, the presence of the Oriental invasive species Stegomyia albopicta (Skuse) (=Aedes albopictus) has never been confirmed in Turkey. Active surveillance for this container-breeding species was carried out using oviposition traps at 15 discrete sites in the towns of Ipsala (n=8 sites), Kesan (n=5) (Edirne District), and Malkara (n=2) (Tekirdag District) in the Thrace region of northwestern Turkey, from May 23 through November 10, 2011. Eggs collected were reared to the fourth larval instar and adult stages where possible to facilitate integrated morphological and molecular species identification. DNA barcodes (658 bp of the mitochondrial cytochrome c oxidase I [COI] gene) were compared with all four potentially invasive Stegomyia species: St. aegypti, St. albopicta, St. cretina, and St. japonica. Sequences generated for samples collected in Thrace Region were herein confirmed as St. albopicta, the first record of this vector species in Turkey. Eggs of St. albopicta were detected in two discrete localities: (1) In the grounds of a restaurant in Kesan (in week 36), and (2) in the customs area of the Turkish-Greek border at Ipsala (in weeks 32 and 38). Multiple detection of St. albopicta eggs indicates the possible establishment of the species in northwestern Turkey. Finding this important disease vector has implications for public health and requires the implementation of active vector monitoring programs and targeted vector suppression strategies to limit the spread of this invasive vector species in Turkey.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Gokumakulapalle, Madhuri; Mei, Ya-Fang, E-mail: ya-fang.mei@umu.se

The use of continuous cell lines derived from the African green monkey kidney (AGMK) has led to major advances in virus vaccine development. However, to date, these cells have not been used to facilitate the creation of human adenoviruses because most human adenoviruses undergo abortive infections in them. Here, we report the susceptibility of AGMK-derived cells to adenovirus 11p (Ad11p) infection. First, we showed that CD46 molecules, which act as receptors for Ad11p, are expressed in AGMK cells. We then monitored Ad11p replication by measuring GFP expression as an indicator of viral transcription. We found that AGMK-derived cells were asmore » capable as carcinoma cells at propagating full-length replication-competent Ad11p (RCAd11p) DNA. Of the AGMK cell lines tested, Vero cells had the greatest capacity for adenovirus production. Thus, AGMK cells can be used to evaluate RCAd11p-mediated gene delivery, and Vero cells can be used for the production of RCAd11pGFP vectors at relatively high yields. - Highlights: • Africa green monkey cell lines were monitored for human adenovirus 11p GFP vector infection. • Human CD46 molecules were detectable in these monkey cell lines. • Adenovirus 11p GFP vector can be propagated in Vero cells increases the safety of Ad11p-based vectors for clinical trials. • To use Vero cells for preparation of Ad11p vector avoids the potential inclusion of oncogenes from tumor cells.« less

Microneedle-mediated immunization of an adenovirus-based malaria vaccine enhances antigen-specific antibody immunity and reduces anti-vector responses compared to the intradermal route

PubMed Central

Carey, John B.; Vrdoljak, Anto; O'Mahony, Conor; Hill, Adrian V. S.; Draper, Simon J.; Moore, Anne C.

2014-01-01

Substantial effort has been placed in developing efficacious recombinant attenuated adenovirus-based vaccines. However induction of immunity to the vector is a significant obstacle to its repeated use. Here we demonstrate that skin-based delivery of an adenovirus-based malaria vaccine, HAdV5-PyMSP142, to mice using silicon microneedles induces equivalent or enhanced antibody responses to the encoded antigen, however it results in decreased anti-vector responses, compared to intradermal delivery. Microneedle-mediated vaccine priming and resultant induction of low anti-vector antibody titres permitted repeated use of the same adenovirus vaccine vector. This resulted in significantly increased antigen-specific antibody responses in these mice compared to ID-treated mice. Boosting with a heterologous vaccine; MVA-PyMSP142 also resulted in significantly greater antibody responses in mice primed with HAdV5-PyMSP142 using MN compared to the ID route. The highest protection against blood-stage malaria challenge was observed when a heterologous route of immunization (MN/ID) was used. Therefore, microneedle-mediated immunization has potential to both overcome some of the logistic obstacles surrounding needle-and-syringe-based immunization as well as to facilitate the repeated use of the same adenovirus vaccine thereby potentially reducing manufacturing costs of multiple vaccines. This could have important benefits in the clinical ease of use of adenovirus-based immunization strategies. PMID:25142082

Molecular Identification of Two Vector Species, Cacopsylla melanoneura and Cacopsylla picta (Hemiptera: Psyllidae), of Apple Proliferation Disease and Further Common Psyllids of Northern Italy.

PubMed

Oettl, Sabine; Schlink, Katja

2015-10-01

The psyllid species Cacopsylla melanoneura (Förster) and Cacopsylla picta (Förster) are vectors of 'Candidatus Phytoplasma mali', the causal agent of apple proliferation, one of the economically most important apple diseases in Europe. Both vectors are present in apple orchards of South Tyrol and Trentino provinces in Northern Italy. As no direct treatment of the disease is possible, monitoring of the psyllids provides information about the vector presence in the orchards and enables targeted control. Thus, fast and reliable identification of the various psyllids occurring in the apple orchards is required. Morphological differentiation is problematic due to extensive resemblance of some psyllid species especially among females and is error-prone for nymphs. Here we present a rapid and cost-effective polymerase chain reaction-restriction fragment length polymorphism method based on the cytochrome c oxidase subunit I region for the molecular identification of the vector species as well as eight further Cacopsylla species present in the orchards. This method was verified through 98.9% consensus with morphologically identified males, through sequencing and subsequent phylogenetic analysis. In case of doubtful morphological identification of females, the method was able to provide a refined species assignment and could also remarkably facilitate the identification of nymphs. © 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.

A method to facilitate and monitor expression of exogenous genes in the rat kidney using plasmid and viral vectors

PubMed Central

Corridon, Peter R.; Rhodes, George J.; Leonard, Ellen C.; Basile, David P.; Gattone, Vincent H.; Bacallao, Robert L.

2013-01-01

Gene therapy has been proposed as a novel alternative to treat kidney disease. This goal has been hindered by the inability to reliably deliver transgenes to target cells throughout the kidney, while minimizing injury. Since hydrodynamic forces have previously shown promising results, we optimized this approach and designed a method that utilizes retrograde renal vein injections to facilitate transgene expression in rat kidneys. We show, using intravital fluorescence two-photon microscopy, that fluorescent albumin and dextrans injected into the renal vein under defined conditions of hydrodynamic pressure distribute broadly throughout the kidney in live animals. We found injection parameters that result in no kidney injury as determined by intravital microscopy, histology, and serum creatinine measurements. Plasmids, baculovirus, and adenovirus vectors, designed to express EGFP, EGFP-actin, EGFP-occludin, EGFP-tubulin, tdTomato-H2B, or RFP-actin fusion proteins, were introduced into live kidneys in a similar fashion. Gene expression was then observed in live and ex vivo kidneys using two-photon imaging and confocal laser scanning microscopy. We recorded widespread fluorescent protein expression lasting more than 1 mo after introduction of transgenes. Plasmid and adenovirus vectors provided gene transfer efficiencies ranging from 50 to 90%, compared with 10–50% using baculovirus. Using plasmids and adenovirus, fluorescent protein expression was observed 1) in proximal and distal tubule epithelial cells; 2) within glomeruli; and 3) within the peritubular interstitium. In isolated kidneys, fluorescent protein expression was observed from the cortex to the papilla. These results provide a robust approach for gene delivery and the study of protein function in live mammal kidneys. PMID:23467422

Application of satellite data in variational analysis for global cyclonic systems

NASA Technical Reports Server (NTRS)

Achtemeier, G. L.

1988-01-01

The goal of the research is a variational data assimilation method that incorporates as dynamical constraints, the primitive equations for a moist, convectively unstable atmosphere and the radiative transfer equation. Variables to be adjusted include the three-dimensional vector wind, height, temperature, and moisture from rawinsonde data, and cloud-wind vectors, moisture, and radiance from satellite data. In order to facilitate thorough analysis of each of the model components, four variational models that divide the problem naturally according to increasing complexity were defined. The research performed during the second year fall into four areas: sensitivity studies involving Model 1; evaluation of Model 2; reformation of Model 1 for greater compatibility with Model 2; development of Model 3 (radiative transfer equation); and making the model more responsive to the observations.

Predicting and mapping malaria under climate change scenarios: the potential redistribution of malaria vectors in Africa.

PubMed

Tonnang, Henri E Z; Kangalawe, Richard Y M; Yanda, Pius Z

2010-04-23

Malaria is rampant in Africa and causes untold mortality and morbidity. Vector-borne diseases are climate sensitive and this has raised considerable concern over the implications of climate change on future disease risk. The problem of malaria vectors (Anopheles mosquitoes) shifting from their traditional locations to invade new zones is an important concern. The vision of this study was to exploit the sets of information previously generated by entomologists, e.g. on geographical range of vectors and malaria distribution, to build models that will enable prediction and mapping the potential redistribution of Anopheles mosquitoes in Africa. The development of the modelling tool was carried out through calibration of CLIMEX parameters. The model helped estimate the potential geographical distribution and seasonal abundance of the species in relation to climatic factors. These included temperature, rainfall and relative humidity, which characterized the living environment for Anopheles mosquitoes. The same parameters were used in determining the ecoclimatic index (EI). The EI values were exported to a GIS package for special analysis and proper mapping of the potential future distribution of Anopheles gambiae and Anophles arabiensis within the African continent under three climate change scenarios. These results have shown that shifts in these species boundaries southward and eastward of Africa may occur rather than jumps into quite different climatic environments. In the absence of adequate control, these predictions are crucial in understanding the possible future geographical range of the vectors and the disease, which could facilitate planning for various adaptation options. Thus, the outputs from this study will be helpful at various levels of decision making, for example, in setting up of an early warning and sustainable strategies for climate change and climate change adaptation for malaria vectors control programmes in Africa.

Insulated piggyBac vectors for insect transgenesis

PubMed Central

Sarkar, Abhimanyu; Atapattu, Asela; Belikoff, Esther J; Heinrich, Jörg C; Li, Xuelei; Horn, Carsten; Wimmer, Ernst A; Scott, Maxwell J

2006-01-01

Background Germ-line transformation of insects is now a widely used method for analyzing gene function and for the development of genetically modified strains suitable for pest control programs. The most widely used transposable element for the germ-line transformation of insects is piggyBac. The site of integration of the transgene can influence gene expression due to the effects of nearby transcription enhancers or silent heterochromatic regions. Position effects can be minimized by flanking a transgene with insulator elements. The scs/scs' and gypsy insulators from Drosophila melanogaster as well as the chicken β-globin HS4 insulator function in both Drosophila and mammalian cells. Results To minimize position effects we have created a set of piggyBac transformation vectors that contain either the scs/scs', gypsy or chicken β-globin HS4 insulators. The vectors contain either fluorescent protein or eye color marker genes and have been successfully used for germ-line transformation of Drosophila melanogaster. A set of the scs/scs' vectors contains the coral reef fluorescent protein marker genes AmCyan, ZsGreen and DsRed that have not been optimized for translation in human cells. These marker genes are controlled by a combined GMR-3xP3 enhancer/promoter that gives particularly strong expression in the eyes. This is also the first report of the use of the ZsGreen and AmCyan reef fluorescent proteins as transformation markers in insects. Conclusion The insulated piggyBac vectors should protect transgenes against position effects and thus facilitate fine control of gene expression in a wide spectrum of insect species. These vectors may also be used for transgenesis in other invertebrate species. PMID:16776846

Gene Therapy for Neurologic Manifestations of Mucopolysaccharidoses

PubMed Central

Wolf, Daniel A.; Banerjee, Sharbani; Hackett, Perry B.; Whitley, Chester B.; McIvor, R. Scott; Low, Walter C.

2015-01-01

Introduction Mucopolysaccharidoses are a family of lysosomal disorders caused by mutations in genes that encode enzymes involved in the catabolism of glycoaminoglycans. These mutations affect multiple organ systems and can be particularly deleterious to the nervous system. At the present time, enzyme replacement therapy and hematopoietic stem-cell therapy are used to treat patients with different forms of these disorders. However, to a great extent the nervous system is not adequately responsive to current therapeutic approaches. Areas Covered Recent advances in gene therapy show great promise for treating mucopolysaccharidoses. This article reviews the current state of the art for routes of delivery in developing genetic therapies for treating the neurologic manifestations of mucopolysaccharidoses. Expert Opinion Gene therapy for treating neurological manifestations of mucopolysaccharidoses can be achieved by intraventricular, intrathecal, intranasal, and systemic administration. The intraventricular route of administration appears to provide the most wide-spread distribution of gene therapy vectors to the brain. The intrathecal route of delivery results in predominant distribution to the caudal areas of the brain while the intranasal route of delivery results in good distribution to the rostral areas of brain. The systemic route of delivery via intravenous delivery can also achieve wide spread delivery to the CNS, however, the distribution to the brain is greatly dependent on the vector system. Intravenous delivery using lentiviral vectors appear to be less effective than adeno-associated viral (AAV) vectors. Moreover, some subtypes of AAV vectors are more effective than others in crossing the blood-brain-barrier. In summary, the recent advances in gene vector technology and routes of delivery to the CNS will facilitate the clinical translation of gene therapy for the treatment of the neurological manifestations of mucopolysaccharidoses. PMID:25510418

Virus immobilization on biomaterial scaffolds through biotin-avidin interaction for improving bone regeneration.

PubMed

Hu, Wei-Wen; Wang, Zhuo; Krebsbach, Paul H

2016-02-01

To spatially control therapeutic gene delivery for potential tissue engineering applications, a biotin-avidin interaction strategy was applied to immobilize viral vectors on biomaterial scaffolds. Both adenoviral vectors and gelatin sponges were biotinylated and avidin was applied to link them in a virus-biotin-avidin-biotin-material (VBABM) arrangement. The tethered viral particles were stably maintained within scaffolds and SEM images illustrated that viral particles were evenly distributed in three-dimensional (3D) gelatin sponges. An in vivo study demonstrated that transgene expression was restricted to the implant sites only and transduction efficiency was improved using this conjugation method. For an orthotopic bone regeneration model, adenovirus encoding BMP-2 (AdBMP2) was immobilized to gelatin sponges before implanting into critical-sized bone defects in rat calvaria. Compared to gelatin sponges with AdBMP2 loaded in a freely suspended form, the VBABM method enhanced gene transfer and bone regeneration was significantly improved. These results suggest that biotin-avidin immobilization of viral vectors to biomaterial scaffolds may be an effective strategy to facilitate tissue regeneration. Copyright © 2013 John Wiley & Sons, Ltd.

Targeting vector construction through recombineering.

PubMed

Malureanu, Liviu A

2011-01-01

Gene targeting in mouse embryonic stem cells is an essential, yet still very expensive and highly time-consuming, tool and method to study gene function at the organismal level or to create mouse models of human diseases. Conventional cloning-based methods have been largely used for generating targeting vectors, but are hampered by a number of limiting factors, including the variety and location of restriction enzymes in the gene locus of interest, the specific PCR amplification of repetitive DNA sequences, and cloning of large DNA fragments. Recombineering is a technique that exploits the highly efficient homologous recombination function encoded by λ phage in Escherichia coli. Bacteriophage-based recombination can recombine homologous sequences as short as 30-50 bases, allowing manipulations such as insertion, deletion, or mutation of virtually any genomic region. The large availability of mouse genomic bacterial artificial chromosome (BAC) libraries covering most of the genome facilitates the retrieval of genomic DNA sequences from the bacterial chromosomes through recombineering. This chapter describes a successfully applied protocol and aims to be a detailed guide through the steps of generation of targeting vectors through recombineering.

Microplastics as vectors for environmental contaminants: Exploring sorption, desorption, and transfer to biota.

PubMed

Hartmann, Nanna B; Rist, Sinja; Bodin, Julia; Jensen, Louise Hs; Schmidt, Stine N; Mayer, Philipp; Meibom, Anders; Baun, Anders

2017-05-01

The occurrence and effects of microplastics (MPs) in the aquatic environment are receiving increasing attention. In addition to their possible direct adverse effects on biota, the potential role of MPs as vectors for hydrophobic organic chemicals (HOCs), compared to natural pathways, is a topic of much debate. It is evident, however, that temporal and spatial variations of MP occurrence do (and will) occur. To further improve the estimations of the role of MPs as vectors for HOC transfer into biota under varying MP concentrations and environmental conditions, it is important to identify and understand the governing processes. Here, we explore HOC sorption to and desorption from MPs and the underlying principles for their interactions. We discuss intrinsic and extrinsic parameters influencing these processes and focus on the importance of the exposure route for diffusive mass transfer. Also, we outline research needed to fill knowledge gaps and improve model-based calculations of MP-facilitated HOC transfer in the environment. Integr Environ Assess Manag 2017;13:488-493. © 2017 SETAC. © 2017 SETAC.

Seasonal dynamics of thrips (Thrips tabaci) (Thysanoptera: Thripidae) transmitters of iris yellow spot virus: a serious viral pathogen of onion bulb and seed crops.

PubMed

Bag, Sudeep; Rondon, Silvia I; Druffel, Keri L; Riley, David G; Pappu, Hanu R

2014-02-01

Thrips-transmitted Iris yellow spot virus (IYSV) is an important economic constraint to the production of bulb and seed onion crops in the United States and many other parts of the world. Because the virus is exclusively spread by thrips, the ability to rapidly detect the virus in thrips vectors would facilitate studies on the role of thrips in virus epidemiology, and thus formulation of better vector management strategies. Using a polyclonal antiserum produced against the recombinant, Escherichia coli-expressed nonstructural protein coded by the small (S) RNA of IYSV, an enzyme linked immunosorbent assay was developed for detecting IYSV in individual as well as groups of adult thrips. The approach enabled estimating the proportion of potential thrips transmitters in a large number of field-collected thrips collected from field-grown onion plants. Availability of a practical and inexpensive test to identify viruliferous thrips would be useful in epidemiological studies to better understand the role of thrips vectors in outbreaks of this economically important virus of onion.

The neural network classification of false killer whale (Pseudorca crassidens) vocalizations.

PubMed

Murray, S O; Mercado, E; Roitblat, H L

1998-12-01

This study reports the use of unsupervised, self-organizing neural network to categorize the repertoire of false killer whale vocalizations. Self-organizing networks are capable of detecting patterns in their input and partitioning those patterns into categories without requiring that the number or types of categories be predefined. The inputs for the neural networks were two-dimensional characterization of false killer whale vocalization, where each vocalization was characterized by a sequence of short-time measurements of duty cycle and peak frequency. The first neural network used competitive learning, where units in a competitive layer distributed themselves to recognize frequently presented input vectors. This network resulted in classes representing typical patterns in the vocalizations. The second network was a Kohonen feature map which organized the outputs topologically, providing a graphical organization of pattern relationships. The networks performed well as measured by (1) the average correlation between the input vectors and the weight vectors for each category, and (2) the ability of the networks to classify novel vocalizations. The techniques used in this study could easily be applied to other species and facilitate the development of objective, comprehensive repertoire models.

Cracking Open the Curriculum: Lessons from a Community College

ERIC Educational Resources Information Center

Hanson, Chad; Amelotte, Patrick

2013-01-01

Community colleges comprise the largest single sector of the US higher education network. Forty percent of undergraduates attend one of our two-year schools. Some estimates suggest that, since the turnover is quicker than on four-year campuses, two-thirds of the students who attend a college at all attend a community college. For many of those…

Ultrafast Direct Modulation of a Single-Mode Photonic Crystal Nanocavity Light-Emitting Diode

DTIC Science & Technology

2011-11-15

nanocavity laser with world record low threshold of 208 nW based on a lateral p-i-n junction defined by ion implantation in gallium arsenide6. This...recombination effects are mini- mized. In contrast, at room temperature, thermal excitation of car- riers depopulates the quantum dots much quicker than does Pur

Computer Adaptive Testing, Big Data and Algorithmic Approaches to Education

ERIC Educational Resources Information Center

Thompson, Greg

2017-01-01

This article critically considers the promise of computer adaptive testing (CAT) and digital data to provide better and quicker data that will improve the quality, efficiency and effectiveness of schooling. In particular, it uses the case of the Australian NAPLAN test that will become an online, adaptive test from 2016. The article argues that…

Analog design of a new neural network for optical character recognition.

PubMed

Morns, I P; Dlay, S S

1999-01-01

An electronic circuit is presented for a new type of neural network, which gives a recognition rate of over 100 kHz. The network is used to classify handwritten numerals, presented as Fourier and wavelet descriptors, and has been shown to train far quicker than the popular backpropagation network while maintaining classification accuracy.

Teaching with an Ever-Spinning Web: Instructional Uses of Web 2.0

ERIC Educational Resources Information Center

Teahan, John

2006-01-01

In little more than a decade, the Web has become an essential medium for information, entertainment, and commerce, making its adoption among consumers quicker than any other medium before it. The majority of American classrooms have become connected to the Internet in the same short time span. Previously, science or mathematics students might have…

Integrating CALL in ESOL Classrooms: Understanding Teachers' Perspectives and Meeting Students' Needs

ERIC Educational Resources Information Center

Nia, Sara Farshad; Davey, Ronnie

2014-01-01

In line with the needs of the 21st century learners and dramatic improvements in schools' technological infrastructures, it is expected that the integration of digital tools into language learning courses would take a quicker pace and a smoother path. However, current research indicates that although this might be the case for foreign language…

Foundations of Neuromorphic Computing

DTIC Science & Technology

2013-05-01

make informed decisions quicker than our adversaries. 2.0 INTRODUCTION The increasing resolution and speed of today’s advanced sensor ...limited information about the location, access to global positioning satellite information (GPS) to aid in navigation is impeded, and communications...more autonomous capability. This is where neuromorphic computing and other bio -inspired technologies for SWaP constrained environments can play a

The Utilization of a Clinical Decision Support System to Manage Adult Type 2 Diabetes: A Correlational Study

ERIC Educational Resources Information Center

Faught, I. Charie

2012-01-01

While the Institute of Medicine (2001) has promoted health information technology to improve the process of care such as compliance with clinical practice guidelines and quicker access to clinical information, diagnostic tests, and treatment results, very little was known about how a clinical decision support system can contribute to diabetes…

78 FR 16823 - Proposed Establishment of Class E Airspace; Tuba City, AZ

Federal Register 2010, 2011, 2012, 2013, 2014

2013-03-19

...., long. 109[deg]35'55'' W.; to lat. 37[deg]04'41'' N., long. 109[deg]38'16'' W.; to lat. 36[deg]57'10'' N...., long. 110[deg]38'05'' W.; to lat. 39[deg]03'55'' N., long. 110[deg]37'49'' W.; thence to the point of... Air Navigational Aid (VORTAC), Tuba City, AZ to facilitate vectoring of Instrument Flight Rules (IFR...

Back to BAC: The Use of Infectious Clone Technologies for Viral Mutagenesis

PubMed Central

Hall, Robyn N.; Meers, Joanne; Fowler, Elizabeth; Mahony, Timothy

2012-01-01

Bacterial artificial chromosome (BAC) vectors were first developed to facilitate the propagation and manipulation of large DNA fragments in molecular biology studies for uses such as genome sequencing projects and genetic disease models. To facilitate these studies, methodologies have been developed to introduce specific mutations that can be directly applied to the mutagenesis of infectious clones (icBAC) using BAC technologies. This has resulted in rapid identification of gene function and expression at unprecedented rates. Here we review the major developments in BAC mutagenesis in vitro. This review summarises the technologies used to construct and introduce mutations into herpesvirus icBAC. It also explores developing technologies likely to provide the next leap in understanding these important viruses. PMID:22470833

Comparison of scoliosis measurements based on three-dimensional vertebra vectors and conventional two-dimensional measurements: advantages in evaluation of prognosis and surgical results.

PubMed

Illés, Tamás; Somoskeöy, Szabolcs

2013-06-01

A new concept of vertebra vectors based on spinal three-dimensional (3D) reconstructions of images from the EOS system, a new low-dose X-ray imaging device, was recently proposed to facilitate interpretation of EOS 3D data, especially with regard to horizontal plane images. This retrospective study was aimed at the evaluation of the spinal layout visualized by EOS 3D and vertebra vectors before and after surgical correction, the comparison of scoliotic spine measurement values based on 3D vertebra vectors with measurements using conventional two-dimensional (2D) methods, and an evaluation of horizontal plane vector parameters for their relationship with the magnitude of scoliotic deformity. 95 patients with adolescent idiopathic scoliosis operated according to the Cotrel-Dubousset principle were subjected to EOS X-ray examinations pre- and postoperatively, followed by 3D reconstructions and generation of vertebra vectors in a calibrated coordinate system to calculate vector coordinates and parameters, as published earlier. Differences in values of conventional 2D Cobb methods and methods based on vertebra vectors were evaluated by means comparison T test and relationship of corresponding parameters was analysed by bivariate correlation. Relationship of horizontal plane vector parameters with the magnitude of scoliotic deformities and results of surgical correction were analysed by Pearson correlation and linear regression. In comparison to manual 2D methods, a very close relationship was detectable in vertebra vector-based curvature data for coronal curves (preop r 0.950, postop r 0.935) and thoracic kyphosis (preop r 0.893, postop r 0.896), while the found small difference in L1-L5 lordosis values (preop r 0.763, postop r 0.809) was shown to be strongly related to the magnitude of corresponding L5 wedge. The correlation analysis results revealed strong correlation between the magnitude of scoliosis and the lateral translation of apical vertebra in horizontal plane. The horizontal plane coordinates of the terminal and initial points of apical vertebra vectors represent this (r 0.701; r 0.667). Less strong correlation was detected in the axial rotation of apical vertebras and the magnitudes of the frontal curves (r 0.459). Vertebra vectors provide a key opportunity to visualize spinal deformities in all three planes simultaneously. Measurement methods based on vertebral vectors proved to be just as accurate and reliable as conventional measurement methods for coronal and sagittal plane parameters. In addition, the horizontal plane display of the curves can be studied using the same vertebra vectors. Based on the vertebra vectors data, during the surgical treatment of spinal deformities, the diminution of the lateral translation of the vertebras seems to be more important in the results of the surgical correction than the correction of the axial rotation.

Self-entanglement of long linear DNA vectors using transient non-B-DNA attachment points: a new concept for improvement of non-viral therapeutic gene delivery.

PubMed

Tolmachov, Oleg E

2012-05-01

The cell-specific and long-term expression of therapeutic transgenes often requires a full array of native gene control elements including distal enhancers, regulatory introns and chromatin organisation sequences. The delivery of such extended gene expression modules to human cells can be accomplished with non-viral high-molecular-weight DNA vectors, in particular with several classes of linear DNA vectors. All high-molecular-weight DNA vectors are susceptible to damage by shear stress, and while for some of the vectors the harmful impact of shear stress can be minimised through the transformation of the vectors to compact topological configurations by supercoiling and/or knotting, linear DNA vectors with terminal loops or covalently attached terminal proteins cannot be self-compacted in this way. In this case, the only available self-compacting option is self-entangling, which can be defined as the folding of single DNA molecules into a configuration with mutual restriction of molecular motion by the individual segments of bent DNA. A negatively charged phosphate backbone makes DNA self-repulsive, so it is reasonable to assume that a certain number of 'sticky points' dispersed within DNA could facilitate the entangling by bringing DNA segments into proximity and by interfering with the DNA slipping away from the entanglement. I propose that the spontaneous entanglement of vector DNA can be enhanced by the interlacing of the DNA with sites capable of mutual transient attachment through the formation of non-B-DNA forms, such as interacting cruciform structures, inter-segment triplexes, slipped-strand DNA, left-handed duplexes (Z-forms) or G-quadruplexes. It is expected that the non-B-DNA based entanglement of the linear DNA vectors would consist of the initial transient and co-operative non-B-DNA mediated binding events followed by tight self-ensnarement of the vector DNA. Once in the nucleoplasm of the target human cells, the DNA can be disentangled by type II topoisomerases. The technology for such self-entanglement can be an avenue for the improvement of gene delivery with high-molecular-weight naked DNA using therapeutically important methods associated with considerable shear stress. Priority applications include in vivo muscle electroporation and sonoporation for Duchenne muscular dystrophy patients, aerosol inhalation to reach the target lung cells of cystic fibrosis patients and bio-ballistic delivery to skin melanomas with the vector DNA adsorbed on gold or tungsten projectiles. Copyright © 2012 Elsevier Ltd. All rights reserved.

Bioreducible Fluorinated Peptide Dendrimers Capable of Circumventing Various Physiological Barriers for Highly Efficient and Safe Gene Delivery.

PubMed

Cai, Xiaojun; Jin, Rongrong; Wang, Jiali; Yue, Dong; Jiang, Qian; Wu, Yao; Gu, Zhongwei

2016-03-09

Polymeric vectors have shown great promise in the development of safe and efficient gene delivery systems; however, only a few have been developed in clinical settings due to poor transport across multiple physiological barriers. To address this issue and promote clinical translocation of polymeric vectors, a new type of polymeric vector, bioreducible fluorinated peptide dendrimers (BFPDs), was designed and synthesized by reversible cross-linking of fluorinated low generation peptide dendrimers. Through masterly integration all of the features of reversible cross-linking, fluorination, and polyhedral oligomeric silsesquioxane (POSS) core-based peptide dendrimers, this novel vector exhibited lots of unique features, including (i) inactive surface to resist protein interactions; (ii) virus-mimicking surface topography to augment cellular uptake; (iii) fluorination-mediated efficient cellular uptake, endosome escape, cytoplasm trafficking, and nuclear entry, and (iv) disulfide-cleavage-mediated polyplex disassembly and DNA release that allows efficient DNA transcription. Noteworthy, all of these features are functionally important and can synergistically facilitate DNA transport from solution to the nucleus. As a consequences, BFPDs showed excellent gene transfection efficiency in several cell lines (∼95% in HEK293 cells) and superior biocompatibility compared with polyethylenimine (PEI). Meanwhile BFPDs provided excellent serum resistance in gene delivery. More importantly, BFPDs offer considerable in vivo gene transfection efficiency (in muscular tissues and in HepG2 tumor xenografts), which was approximately 77-fold higher than that of PEI in luciferase activity. These results suggest bioreducible fluorinated peptide dendrimers are a new class of highly efficient and safe gene delivery vectors and should be used in clinical settings.

Generation of monoclonal antibodies to vertebrate albumins for analysis of arthropod blood meals.

PubMed

Schwab, Lori Kae; Nardi, James B; Holly, Theresa; Wang, Liping; Frye, Janie; Novak, Robert J

2011-06-01

An immunoassay using monoclonal antibodies (MAbs) that are specific for different vertebrate taxa (from class to species) has been developed that simplifies and facilitates analysis of vertebrate blood meals from arthropod vectors. The MAbs have been prepared against the single protein albumin, the most abundant protein in vertebrate sera. A panel of these antibodies has been generated against albumins from 33 species of vertebrates, representing four classes, 15 orders, and 25 families. Immunoreactivity of albumin in mosquito blood meals can be detected as late as 48 h after feeding. Immunoassays with MAbs can be carried out in the field as well as the laboratory. Used in conjunction with nucleic acid assays or used alone with an appropriate assortment of antibodies, the assay is simple, sensitive, and unambiguous. © 2011 The Society for Vector Ecology.

Tick-Borne Viruses.

PubMed

Shi, Junming; Hu, Zhihong; Deng, Fei; Shen, Shu

2018-02-01

Ticks are important vectors for the transmission of pathogens including viruses. The viruses carried by ticks also known as tick-borne viruses (TBVs), contain a large group of viruses with diverse genetic properties and are concluded in two orders, nine families, and at least 12 genera. Some members of the TBVs are notorious agents causing severe diseases with high mortality rates in humans and livestock, while some others may pose risks to public health that are still unclear to us. Herein, we review the current knowledge of TBVs with emphases on the history of virus isolation and identification, tick vectors, and potential pathogenicity to humans and animals, including assigned species as well as the recently discovered and unassigned species. All these will promote our understanding of the diversity of TBVs, and will facilitate the further investigation of TBVs in association with both ticks and vertebrate hosts.

Folic acid-decorated polyamidoamine dendrimer mediates selective uptake and high expression of genes in head and neck cancer cells.

PubMed

Xu, Leyuan; Kittrell, Shannon; Yeudall, W Andrew; Yang, Hu

2016-11-01

Folic acid (FA)-decorated polyamidoamine dendrimer G4 (G4-FA) was synthesized and studied for targeted delivery of genes to head and neck cancer cells expressing high levels of folate receptors (FRs). Cellular uptake, targeting specificity, cytocompatibility and transfection efficiency were evaluated. G4-FA competes with free FA for the same binding site. G4-FA facilitates the cellular uptake of DNA plasmids in a FR-dependent manner and selectively delivers plasmids to FR-high cells, leading to enhanced gene expression. G4-FA is a suitable vector to deliver genes selectively to head and neck cancer cells. The fundamental understandings of G4-FA as a vector and its encouraging transfection results for head and neck cancer cells provided support for its further testing in vivo.

Steganalysis of recorded speech

NASA Astrophysics Data System (ADS)

Johnson, Micah K.; Lyu, Siwei; Farid, Hany

2005-03-01

Digital audio provides a suitable cover for high-throughput steganography. At 16 bits per sample and sampled at a rate of 44,100 Hz, digital audio has the bit-rate to support large messages. In addition, audio is often transient and unpredictable, facilitating the hiding of messages. Using an approach similar to our universal image steganalysis, we show that hidden messages alter the underlying statistics of audio signals. Our statistical model begins by building a linear basis that captures certain statistical properties of audio signals. A low-dimensional statistical feature vector is extracted from this basis representation and used by a non-linear support vector machine for classification. We show the efficacy of this approach on LSB embedding and Hide4PGP. While no explicit assumptions about the content of the audio are made, our technique has been developed and tested on high-quality recorded speech.

Library Resources for Bac End Sequencing. Final Technical Report

DOE Office of Scientific and Technical Information (OSTI.GOV)

Pieter J. de Jong

2000-10-01

Studies directed towards the specific aims outlined for this research award are summarized. The RPCI II Human Bac Library has been expanded by the addition of 6.9-fold genomic coverage. This segment has been generated from a MBOI partial digest of the same anonymous donor DNA used for the rest of the library. A new cloning vector, pTARBAC1, has been constructed and used in the construction of RPCI-II segment 5. This new cloning vector provides a new strategy in identifying targeted genomic regions and will greatly facilitate a large-scale analysis for positional cloning. A new maleCS7BC/6J mouse BAC library has beenmore » constructed. RPCI-23 contain 576 plates (approx 210,000 clones) and represents approximately 11-fold coverage of the mouse genome.« less

Spatial data analytics on heterogeneous multi- and many-core parallel architectures using python

USGS Publications Warehouse

Laura, Jason R.; Rey, Sergio J.

2017-01-01

Parallel vector spatial analysis concerns the application of parallel computational methods to facilitate vector-based spatial analysis. The history of parallel computation in spatial analysis is reviewed, and this work is placed into the broader context of high-performance computing (HPC) and parallelization research. The rise of cyber infrastructure and its manifestation in spatial analysis as CyberGIScience is seen as a main driver of renewed interest in parallel computation in the spatial sciences. Key problems in spatial analysis that have been the focus of parallel computing are covered. Chief among these are spatial optimization problems, computational geometric problems including polygonization and spatial contiguity detection, the use of Monte Carlo Markov chain simulation in spatial statistics, and parallel implementations of spatial econometric methods. Future directions for research on parallelization in computational spatial analysis are outlined.

Geminiviruses for biotechnology: the art of parasite taming.

PubMed

Lozano-Durán, Rosa

2016-04-01

Viruses are intracellular pathogens that have evolved efficient strategies for replication and expression of their proteins in the host cells. Geminiviruses - plant viruses with small circular single-stranded DNA genomes - effectively manipulate plant cell processes for viral functions, entailing great potential for biotechnological applications. This potentiality has been realized in the form of protein expression and gene-silencing vectors, and, more recently, vectors for genome editing - a technology that these viruses seem particularly well-suited to facilitate. This insight offers an overview of the biological properties of geminiviruses, with emphasis on those leveraging development of geminivirus-based replicons. It illustrates the basis for engineering geminivirus-based replicons and their applications. Furthermore, it discusses the reported use and future perspectives of geminivirus-based replicons for genome editing. © 2015 The Authors. New Phytologist © 2015 New Phytologist Trust.

NMR-based urine analysis in rats: prediction of proximal tubule kidney toxicity and phospholipidosis.

PubMed

Lienemann, Kai; Plötz, Thomas; Pestel, Sabine

2008-01-01

The aim of safety pharmacology is early detection of compound-induced side-effects. NMR-based urine analysis followed by multivariate data analysis (metabonomics) identifies efficiently differences between toxic and non-toxic compounds; but in most cases multiple administrations of the test compound are necessary. We tested the feasibility of detecting proximal tubule kidney toxicity and phospholipidosis with metabonomics techniques after single compound administration as an early safety pharmacology approach. Rats were treated orally, intravenously, inhalatively or intraperitoneally with different test compounds. Urine was collected at 0-8 h and 8-24 h after compound administration, and (1)H NMR-patterns were recorded from the samples. Variation of post-processing and feature extraction methods led to different views on the data. Support Vector Machines were trained on these different data sets and then aggregated as experts in an Ensemble. Finally, validity was monitored with a cross-validation study using a training, validation, and test data set. Proximal tubule kidney toxicity could be predicted with reasonable total classification accuracy (85%), specificity (88%) and sensitivity (78%). In comparison to alternative histological studies, results were obtained quicker, compound need was reduced, and very importantly fewer animals were needed. In contrast, the induction of phospholipidosis by the test compounds could not be predicted using NMR-based urine analysis or the previously published biomarker PAG. NMR-based urine analysis was shown to effectively predict proximal tubule kidney toxicity after single compound administration in rats. Thus, this experimental design allows early detection of toxicity risks with relatively low amounts of compound in a reasonably short period of time.

Paint it Black: One-Step Etch Cuts Solar Cell Costs - Continuum Magazine

Science.gov Websites

quicker, cheaper way to produce large volumes of high-performance PV devices. Cost is a major obstacle for propel PV toward cost-competitiveness. A New Approach to Antireflection Any light reflected from a solar etching large pyramids into the cell surface, add considerable cost to a solar cell-and they succeed only

Two vs. One Keep Away: A Simple Game of Beginning Tactics

ERIC Educational Resources Information Center

Belka, David E.

2005-01-01

In the last decade or so, there has been a shift toward small-sided games played in smaller spaces, with limited rules and an emphasis on tactics. The goal of the current instruction is to use less complex game settings to develop players who then learn to make more appropriate and quicker tactical decisions. In this article, the author presents…

Quality Assurance for Rapid Airfield Construction

DTIC Science & Technology

2008-05-01

necessary to conduct a volume-replacement density test for in-place soil. This density test, which was developed during this investigation, involves...the test both simpler and quicker. The Clegg hammer results are the primary means of judging compaction; thus, the requirements for density tests are...minimized through a stepwise acceptance procedure. Statistical criteria for evaluating Clegg hammer and density measurements are also included

Employing Replay Connectors for SIEM Operator Education

DTIC Science & Technology

2013-09-01

BLANK xiii LIST OF ACRONYMS AND ABBREVIATIONS CORR Correlation Optimized Retention and Retrieval CII Critical Information Infrastructure GLBA...vast distances is now quicker and easier with the advancement in mobile computing devices and more ubiquitous connectivity and bandwidth. As a result...breakdown of the Critical Information Infrastructure (CII) is one of the core risks facing the international economy. The World Economic Forum

An Electronic Indicator for Angular Velocity and Acceleration

DTIC Science & Technology

1944-08-01

n’cl | hce (f -l) r>iitlil rl Id, ~3/.ui., , JO. f NACÄ •ft! to AUG8 1947 WASHINGTON riACA WART1MK REPORTS are reprints of papere -r’./inally...ar top suggested tijo aeed for a stapler ml quicker method of obtaining data fron the machine than that provided by the manufacturer, tu the four

A Study of the Utility of Results from the 1992 Trial State Assessment (TSA) in Reading for State-Level Administrators of Assessment.

ERIC Educational Resources Information Center

Bullock, Cheryl Davis; DeStefano, Lizanne

1998-01-01

The usefulness of the 1992 TSA in reading was studied using interviews from 26 state directors of assessment. Perceptions about TSA credibility and orientation of test components and current use of TSA results were examined. The directors suggested involving more teachers in assessment, modifying descriptors, disseminating results quicker, and…

21st Century Readiness Skills for Career and Technical Education

ERIC Educational Resources Information Center

Bennett, Tyrone L.

2012-01-01

The 21st century has embraced the technological age by storm and has approached us quicker than most have expected, leaving many still stuck in the 20th century. Technical careers require very skilled workers and their education has sprung as a strong vehicle all over the country but still most of our employed citizens, especially the youth, have…

Delays in Global Disease Outbreak Responses: Lessons from H1N1, Ebola, and Zika

PubMed Central

Silverberg, Sarah L.

2018-01-01

In global disease outbreaks, there are significant time delays between the source of an outbreak and collective action. Some delay is necessary, but recent delays have been extended by insufficient surveillance capacity and time-consuming efforts to mobilize action. Three public health emergencies of international concern (PHEICs)—H1N1, Ebola, and Zika—allow us to identify and compare sources of delays and consider seven hypotheses about what influences the length of delays. These hypotheses can then motivate further research that empirically tests them. The three PHEICs suggest that deferred global mobilization is a greater source of delay than is poor surveillance capacity. These case study outbreaks support hypotheses that we see quicker responses for novel diseases when outbreaks do not coincide with holidays and when US citizens are infected. They do not support hypotheses that we see quicker responses for more severe outbreaks or those that threaten larger numbers of people. Better understanding the reason for delays can help target policy interventions and identify the kind of global institutional changes needed to reduce the spread and severity of future PHEICs. PMID:29345996

A Bioinformatics Approach for Integrated Transcriptomic and Proteomic Comparative Analyses of Model and Non-sequenced Anopheline Vectors of Human Malaria Parasites*

PubMed Central

Mohien, Ceereena Ubaida; Colquhoun, David R.; Mathias, Derrick K.; Gibbons, John G.; Armistead, Jennifer S.; Rodriguez, Maria C.; Rodriguez, Mario Henry; Edwards, Nathan J.; Hartler, Jürgen; Thallinger, Gerhard G.; Graham, David R.; Martinez-Barnetche, Jesus; Rokas, Antonis; Dinglasan, Rhoel R.

2013-01-01

Malaria morbidity and mortality caused by both Plasmodium falciparum and Plasmodium vivax extend well beyond the African continent, and although P. vivax causes between 80 and 300 million severe cases each year, vivax transmission remains poorly understood. Plasmodium parasites are transmitted by Anopheles mosquitoes, and the critical site of interaction between parasite and host is at the mosquito's luminal midgut brush border. Although the genome of the “model” African P. falciparum vector, Anopheles gambiae, has been sequenced, evolutionary divergence limits its utility as a reference across anophelines, especially non-sequenced P. vivax vectors such as Anopheles albimanus. Clearly, technologies and platforms that bridge this substantial scientific gap are required in order to provide public health scientists with key transcriptomic and proteomic information that could spur the development of novel interventions to combat this disease. To our knowledge, no approaches have been published that address this issue. To bolster our understanding of P. vivax–An. albimanus midgut interactions, we developed an integrated bioinformatic-hybrid RNA-Seq-LC-MS/MS approach involving An. albimanus transcriptome (15,764 contigs) and luminal midgut subproteome (9,445 proteins) assembly, which, when used with our custom Diptera protein database (685,078 sequences), facilitated a comparative proteomic analysis of the midgut brush borders of two important malaria vectors, An. gambiae and An. albimanus. PMID:23082028

A bioinformatics approach for integrated transcriptomic and proteomic comparative analyses of model and non-sequenced anopheline vectors of human malaria parasites.

PubMed

Ubaida Mohien, Ceereena; Colquhoun, David R; Mathias, Derrick K; Gibbons, John G; Armistead, Jennifer S; Rodriguez, Maria C; Rodriguez, Mario Henry; Edwards, Nathan J; Hartler, Jürgen; Thallinger, Gerhard G; Graham, David R; Martinez-Barnetche, Jesus; Rokas, Antonis; Dinglasan, Rhoel R

2013-01-01

Malaria morbidity and mortality caused by both Plasmodium falciparum and Plasmodium vivax extend well beyond the African continent, and although P. vivax causes between 80 and 300 million severe cases each year, vivax transmission remains poorly understood. Plasmodium parasites are transmitted by Anopheles mosquitoes, and the critical site of interaction between parasite and host is at the mosquito's luminal midgut brush border. Although the genome of the "model" African P. falciparum vector, Anopheles gambiae, has been sequenced, evolutionary divergence limits its utility as a reference across anophelines, especially non-sequenced P. vivax vectors such as Anopheles albimanus. Clearly, technologies and platforms that bridge this substantial scientific gap are required in order to provide public health scientists with key transcriptomic and proteomic information that could spur the development of novel interventions to combat this disease. To our knowledge, no approaches have been published that address this issue. To bolster our understanding of P. vivax-An. albimanus midgut interactions, we developed an integrated bioinformatic-hybrid RNA-Seq-LC-MS/MS approach involving An. albimanus transcriptome (15,764 contigs) and luminal midgut subproteome (9,445 proteins) assembly, which, when used with our custom Diptera protein database (685,078 sequences), facilitated a comparative proteomic analysis of the midgut brush borders of two important malaria vectors, An. gambiae and An. albimanus.

A Comprehensive Optimization Strategy for Real-time Spatial Feature Sharing and Visual Analytics in Cyberinfrastructure

NASA Astrophysics Data System (ADS)

Li, W.; Shao, H.

2017-12-01

For geospatial cyberinfrastructure enabled web services, the ability of rapidly transmitting and sharing spatial data over the Internet plays a critical role to meet the demands of real-time change detection, response and decision-making. Especially for the vector datasets which serve as irreplaceable and concrete material in data-driven geospatial applications, their rich geometry and property information facilitates the development of interactive, efficient and intelligent data analysis and visualization applications. However, the big-data issues of vector datasets have hindered their wide adoption in web services. In this research, we propose a comprehensive optimization strategy to enhance the performance of vector data transmitting and processing. This strategy combines: 1) pre- and on-the-fly generalization, which automatically determines proper simplification level through the introduction of appropriate distance tolerance (ADT) to meet various visualization requirements, and at the same time speed up simplification efficiency; 2) a progressive attribute transmission method to reduce data size and therefore the service response time; 3) compressed data transmission and dynamic adoption of a compression method to maximize the service efficiency under different computing and network environments. A cyberinfrastructure web portal was developed for implementing the proposed technologies. After applying our optimization strategies, substantial performance enhancement is achieved. We expect this work to widen the use of web service providing vector data to support real-time spatial feature sharing, visual analytics and decision-making.

Hypoxia-inducible vascular endothelial growth factor gene therapy using the oxygen-dependent degradation domain in myocardial ischemia.

PubMed

Kim, Hyun Ah; Lim, Soyeon; Moon, Hyung-Ho; Kim, Sung Wan; Hwang, Ki-Chul; Lee, Minhyung; Kim, Sun Hwa; Choi, Donghoon

2010-10-01

A hypoxia-inducible VEGF expression system with the oxygen-dependent degradation (ODD) domain was constructed and tested to be used in gene therapy for ischemic myocardial disease. Luciferase and VEGF expression vector systems were constructed with or without the ODD domain: pEpo-SV-Luc (or pEpo-SV-VEGF) and pEpo-SV-Luc-ODD (or pEpo-SV-VEGF-ODD). In vitro gene expression efficiency of each vector type was evaluated in HEK 293 cells under both hypoxic and normoxic conditions. The amount of VEGF protein was estimated by ELISA. The VEGF expression vectors with or without the ODD domain were injected into ischemic rat myocardium. Fibrosis, neovascularization, and cardiomyocyte apoptosis were assessed using Masson's trichrome staining, α-smooth muscle actin (α-SMA) immunostaining, and the TUNEL assay, respectively. The plasmid vectors containing ODD significantly improved the expression level of VEGF protein in hypoxic conditions. The enhancement of VEGF protein production was attributed to increased protein stability due to oxygen deficiency. In a rat model of myocardial ischemia, the pEpo-SV-VEGF-ODD group exhibited less myocardial fibrosis, higher microvessel density, and less cardiomyocyte apoptosis compared to the control groups (saline and pEpo-SV-VEGF treatments). An ODD-mediated VEGF expression system that facilitates VEGF-production under hypoxia may be useful in the treatment of ischemic heart disease.

Development of Third-generation Cocal Envelope Producer Cell Lines for Robust Lentiviral Gene Transfer into Hematopoietic Stem Cells and T-cells.

PubMed

Humbert, Olivier; Gisch, Don W; Wohlfahrt, Martin E; Adams, Amie B; Greenberg, Phil D; Schmitt, Tom M; Trobridge, Grant D; Kiem, Hans-Peter

2016-08-01

Lentiviral vectors (LVs) pseudotyped with vesicular stomatitis virus envelope glycoprotein (VSV-G) have demonstrated great promise in gene therapy trials employing hematopoietic stem cell and T-cells. The VSV-G envelope confers broad tropism and stability to the vector but is toxic when constitutively expressed, which has impeded efforts to generate stable producer cell lines. We previously showed that cocal pseudotyped LVs offer an excellent alternative to VSV-G vectors because of their broad tropism and resistance to human serum inactivation. In this study, we demonstrate that cocal LVs transduce CD34(+) and CD4(+) T-cells more efficiently than VSV-G LVs and share the same receptor(s) for cell entry. 293T-cells stably expressing the cocal envelope produced significantly higher LV titers than VSV-G expressing cells. We developed cocal pseudotyped, third-generation, self-inactivating LV producer cell lines for a GFP reporter and for a WT1 tumor-specific T-cell receptor, which achieved concentrated titers above 10(8) IU/ml and were successfully adapted for growth in suspension, serum-free culture. The resulting LVs were at least as effective as standard LVs in transducing CD34(+) and CD4(+) T-cells. Our stable cocal LV producer cell lines should facilitate the production of large-scale, high titer clinical grade vectors.

Amiloride-enhanced gene transfection of octa-arginine functionalized calcium phosphate nanoparticles

PubMed Central

Tenkumo, Taichi; Kamano, Yuya; Egusa, Hiroshi; Sasaki, Keiichi

2017-01-01

Nanoparticles represent promising gene delivery systems in biomedicine to facilitate prolonged gene expression with low toxicity compared to viral vectors. Specifically, nanoparticles of calcium phosphate (nCaP), the main inorganic component of human bone, exhibit high biocompatibility and good biodegradability and have been reported to have high affinity for protein or DNA, having thus been used as gene transfer vectors. On the other hand, Octa-arginine (R8), which has a high permeability to cell membrane, has been reported to improve intracellular delivery systems. Here, we present an optimized method for nCaP-mediated gene delivery using an octa-arginine (R8)-functionalized nCaP vector containing a marker or functional gene construct. nCaP particle size was between 220–580 nm in diameter and all R8-functionalized nCaPs carried a positive charge. R8 concentration significantly improved nCaP transfection efficiency with high cell compatibility in human mesenchymal stem cells (hMSC) and human osteoblasts (hOB) in particular, suggesting nCaPs as a good option for non-viral vector gene delivery. Furthermore, pre-treatment with different endocytosis inhibitors identified that the endocytic pathway differed among cell lines and functionalized nanoparticles, with amiloride increasing transfection efficiency of R8-functionalized nCaPs in hMSC and hOB. PMID:29145481

Endoscopic and laparoscopic treatment of gastroesophageal reflux.

PubMed

Watson, David I; Immanuel, Arul

2010-04-01

Gastroesophageal reflux is extremely common in Western countries. For selected patients, there is an established role for the surgical treatment of reflux, and possibly an emerging role for endoscopic antireflux procedures. Randomized trials have compared medical versus surgical management, laparoscopic versus open surgery and partial versus total fundoplications. However, the evidence base for endoscopic procedures is limited to some small sham-controlled studies, and cohort studies with short-term follow-up. Laparoscopic fundoplication has been shown to be an effective antireflux operation. It facilitates quicker convalescence and is associated with fewer complications, but has a similar longer term outcome compared with open antireflux surgery. In most randomized trials, antireflux surgery achieves at least as good control of reflux as medical therapy, and these studies support a wider application of surgery for the treatment of moderate-to-severe reflux. Laparoscopic partial fundoplication is an effective surgical procedure with fewer side effects, and it may achieve high rates of patient satisfaction at late follow-up. Many of the early endoscopic antireflux procedures have failed to achieve effective reflux control, and they have been withdrawn from the market. Newer procedures have the potential to fashion a surgical fundoplication. However, at present there is insufficient evidence to establish the safety and efficacy of endoscopic procedures for the treatment of gastroesophageal reflux, and no endoscopic procedure has achieved equivalent reflux control to that achieved by surgical fundoplication.

Failsafe automation of Phase II clinical trial interim monitoring for stopping rules.

PubMed

Day, Roger S

2010-02-01

In Phase II clinical trials in cancer, preventing the treatment of patients on a study when current data demonstrate that the treatment is insufficiently active or too toxic has obvious benefits, both in protecting patients and in reducing sponsor costs. Considerable efforts have gone into experimental designs for Phase II clinical trials with flexible sample size, usually implemented by early stopping rules. The intended benefits will not ensue, however, if the design is not followed. Despite the best intentions, failures can occur for many reasons. The main goal is to develop an automated system for interim monitoring, as a backup system supplementing the protocol team, to ensure that patients are protected. A secondary goal is to stimulate timely recording of patient assessments. We developed key concepts and performance needs, then designed, implemented, and deployed a software solution embedded in the clinical trials database system. The system has been in place since October 2007. One clinical trial tripped the automated monitor, resulting in e-mails that initiated statistician/investigator review in timely fashion. Several essential contributing activities still require human intervention, institutional policy decisions, and institutional commitment of resources. We believe that implementing the concepts presented here will provide greater assurance that interim monitoring plans are followed and that patients are protected from inadequate response or excessive toxicity. This approach may also facilitate wider acceptance and quicker implementation of new interim monitoring algorithms.

Children's accuracy of portion size estimation using digital food images: effects of interface design and size of image on computer screen.

PubMed

Baranowski, Tom; Baranowski, Janice C; Watson, Kathleen B; Martin, Shelby; Beltran, Alicia; Islam, Noemi; Dadabhoy, Hafza; Adame, Su-heyla; Cullen, Karen; Thompson, Debbe; Buday, Richard; Subar, Amy

2011-03-01

To test the effect of image size and presence of size cues on the accuracy of portion size estimation by children. Children were randomly assigned to seeing images with or without food size cues (utensils and checked tablecloth) and were presented with sixteen food models (foods commonly eaten by children) in varying portion sizes, one at a time. They estimated each food model's portion size by selecting a digital food image. The same food images were presented in two ways: (i) as small, graduated portion size images all on one screen or (ii) by scrolling across large, graduated portion size images, one per sequential screen. Laboratory-based with computer and food models. Volunteer multi-ethnic sample of 120 children, equally distributed by gender and ages (8 to 13 years) in 2008-2009. Average percentage of correctly classified foods was 60·3 %. There were no differences in accuracy by any design factor or demographic characteristic. Multiple small pictures on the screen at once took half the time to estimate portion size compared with scrolling through large pictures. Larger pictures had more overestimation of size. Multiple images of successively larger portion sizes of a food on one computer screen facilitated quicker portion size responses with no decrease in accuracy. This is the method of choice for portion size estimation on a computer.

[The development and operation of a package inserts service system for electronic medical records].

PubMed

Yamada, Hidetoshi; Nishimura, Sachiho; Shimamori, Yoshimitsu; Sato, Seiji; Hayase, Yukitoshi

2003-03-01

To promote the appropriate use of pharmaceuticals and to prevent side effects, physicians need package inserts on medicinal drugs as soon as possible. A medicinal drug information service system was established for electronic medical records to speed up and increase the efficiency of package insert communications within a medical institution. Development of this system facilitates access to package inserts by, for example, physicians. The time required to maintain files of package inserts was shortened, and the efficiency of the drug information service increased. As a source of package inserts for this system, package inserts using a standard generalized markup language (SGML) form were used, which are accessible to the public on the homepage of the Organization for Pharmaceutical Safety and Research (OPSR). This study found that a delay occurred in communicating revised package inserts from pharmaceutical companies to the OPSR. Therefore a pharmaceutical department page was set up as part of the homepage of the medical institution for electronic medical records to shorten the delay in the revision of package inserts posted on the medicinal drug information service homepage of the OPSR. The usefulness of this package insert service system for electronic medical records is clear. For more effective use of this system based on the OPSR homepage pharmaceutical companies have been requested to provide quicker updating of package inserts.

Using viral vectors as gene transfer tools (Cell Biology and Toxicology Special Issue: ETCS-UK 1 day meeting on genetic manipulation of cells).

PubMed

Howarth, Joanna L; Lee, Youn Bok; Uney, James B

2010-02-01

In recent years, the development of powerful viral gene transfer techniques has greatly facilitated the study of gene function. This review summarises some of the viral delivery systems routinely used to mediate gene transfer into cell lines, primary cell cultures and in whole animal models. The systems described were originally discussed at a 1-day European Tissue Culture Society (ETCS-UK) workshop that was held at University College London on 1st April 2009. Recombinant-deficient viral vectors (viruses that are no longer able to replicate) are used to transduce dividing and post-mitotic cells, and they have been optimised to mediate regulatable, powerful, long-term and cell-specific expression. Hence, viral systems have become very widely used, especially in the field of neurobiology. This review introduces the main categories of viral vectors, focusing on their initial development and highlighting modifications and improvements made since their introduction. In particular, the use of specific promoters to restrict expression, translational enhancers and regulatory elements to boost expression from a single virion and the development of regulatable systems is described.

Enduring high-efficiency in vivo transfection of neurons with non-viral magnetoparticles in the rat visual cortex for optogenetic applications.

PubMed

Soto-Sánchez, Cristina; Martínez-Navarrete, Gema; Humphreys, Lawrence; Puras, Gustavo; Zarate, Jon; Pedraz, José Luis; Fernández, Eduardo

2015-05-01

This work demonstrates the successful long-term transfection in vivo of a DNA plasmid vector in rat visual cortex neurons using the magnetofection technique. The transfection rates reached values of up to 97% of the neurons after 30days, comparable to those achieved by viral vectors. Immunohistochemical treatment with anti-EGFP antibodies enhanced the detection of the EYFP-channelrhodopsin expression throughout the dendritic trees and cell bodies. These results show that magnetic nanoparticles offer highly efficient and enduring in vivo high-rate transfection in identified neurons of an adult mammalian brain and suggest that the magnetotechnique facilitates the introduction of large functional genetic material like channelrhodopsin with safe non-viral vectors using minimally invasive approaches. Gene therapy may be one of the treatment modalities for neurological diseases in the future. The use of viral transfection remains a concern due to restrictions to the size limit of the genetic material able to be packed, as well as safety issues. In this work, the authors evaluated magnetoplexes as an alternative vehicle. The results showed very promising data in that these nanoparticles could offer high transfection efficiency. Copyright © 2015 Elsevier Inc. All rights reserved.

Agent Collaborative Target Localization and Classification in Wireless Sensor Networks

PubMed Central

Wang, Xue; Bi, Dao-wei; Ding, Liang; Wang, Sheng

2007-01-01

Wireless sensor networks (WSNs) are autonomous networks that have been frequently deployed to collaboratively perform target localization and classification tasks. Their autonomous and collaborative features resemble the characteristics of agents. Such similarities inspire the development of heterogeneous agent architecture for WSN in this paper. The proposed agent architecture views WSN as multi-agent systems and mobile agents are employed to reduce in-network communication. According to the architecture, an energy based acoustic localization algorithm is proposed. In localization, estimate of target location is obtained by steepest descent search. The search algorithm adapts to measurement environments by dynamically adjusting its termination condition. With the agent architecture, target classification is accomplished by distributed support vector machine (SVM). Mobile agents are employed for feature extraction and distributed SVM learning to reduce communication load. Desirable learning performance is guaranteed by combining support vectors and convex hull vectors. Fusion algorithms are designed to merge SVM classification decisions made from various modalities. Real world experiments with MICAz sensor nodes are conducted for vehicle localization and classification. Experimental results show the proposed agent architecture remarkably facilitates WSN designs and algorithm implementation. The localization and classification algorithms also prove to be accurate and energy efficient.

How Many Words Is a Picture Worth? Integrating Visual Literacy in Language Learning with Photographs

ERIC Educational Resources Information Center

Baker, Lottie

2015-01-01

Cognitive research has shown that the human brain processes images quicker than it processes words, and images are more likely than text to remain in long-term memory. With the expansion of technology that allows people from all walks of life to create and share photographs with a few clicks, the world seems to value visual media more than ever…

Accounts receivable cash management: it all happens at the point-of-service.

PubMed

Hajny, T

2000-01-01

If your accounts receivable is not generating the cash your practice needs in a timely and effective manner, look first at your front-end processes. Nine times out of ten, this is where the system is failing. The improvements suggested here will not only generate cash quicker, but also will enhance customer service and lower overall costs by reducing the need for rework.

Alternate method of source preparation for alpha spectrometry: No electrodeposition, no hydrofluoric acid

DOE PAGES

Kurosaki, Hiromu; Mueller, Rebecca J.; Lambert, Susan B.; ...

2016-07-15

An alternate method of preparing actinide alpha counting sources was developed in place of electrodeposition or lanthanide fluoride micro-precipitation. The method uses lanthanide hydroxide micro-precipitation to avoid the use of hazardous hydrofluoric acid. Lastly, it provides a quicker, simpler, and safer way of preparing actinide alpha counting sources in routine, production-type laboratories that process many samples daily.

Malaria Capacity Building in Liberia: The US Navy Joins Forces to Defeat a Deadly Foe

DTIC Science & Technology

2014-01-01

Naval Medical Research Unit No. 3 (NAMRU-3) and the Liberian Institute for Biomedical Research (LIBR) to deliver a two-week Public Health and Vector...3. This facilitated maneu- vering throughout the buildings, Figure 3: Liberians living on EBK military barracks removed their furniture and...with Liberia over four years ago, there have been enormous strides by the Liberians to turn the tide against malaria after long and brutal civil wars

A multisite gateway-based toolkit for targeted gene expression and hairpin RNA silencing in tomato fruits.

PubMed

Estornell, Leandro Hueso; Orzáez, Diego; López-Peña, Lucas; Pineda, Benito; Antón, María Teresa; Moreno, Vicente; Granell, Antonio

2009-04-01

A collection of fruit promoters, reporter genes and protein tags has been constructed in a triple-gateway format, a recombination-based cloning system that facilitates the tandem assembly of three DNA fragments into plant expression vectors. The new pENFRUIT collection includes, among others, the classical tomato-ripening promoters E8 and 2A11 and a set of six new tomato promoters. The new promoter activities were characterized in both transient assays and stable transgenic plants. The range of expression of the new promoters comprises strong (PNH, PLI), medium (PLE, PFF, PHD) and weak (PSN) promoters driving gene expression preferentially in the fruit, and covering a wide range of tissues and developmental stages. Together, a total of 78 possible combinations for the expression of a gene of interest in the fruit, plus a set of five reporters for new promoter analysis, was made available in the current collection. Moreover, the pENFRUIT promoter collection is adaptable to hairpin RNA strategies aimed at tissue/organ-specific gene silencing with only an additional cloning step. The pENFRUIT toolkit broadens the spectrum of promoter activities available for fruit biotechnology and fundamental research, and bypasses technical difficulties of current ligase-dependent cloning techniques in the construction of fruit expression cassettes. The pENFRUIT vector collection is available for the research community in a plasmid repository, facilitating its accessibility.

The crystal structures of two salivary cystatins from the tick Ixodes scapularis and the effect of these inhibitors on the establishment of Borrelia burgdorferi infection in a murine model

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kotsyfakis, Michalis; Horka, Helena; Salat, Jiri

2010-11-17

We have previously demonstrated that two salivary cysteine protease inhibitors from the Borrelia burgdorferi (Lyme disease) vector Ixodes scapularis - namely sialostatins L and L2 - play an important role in tick biology, as demonstrated by the fact that silencing of both sialostatins in tandem results in severe feeding defects. Here we show that sialostatin L2 - but not sialostatin L - facilitates the growth of B. burgdorferi in murine skin. To examine the structural basis underlying these differential effects of the two sialostatins, we have determined the crystal structures of both sialostatin L and L2. This is the firstmore » structural analysis of cystatins from an invertebrate source. Sialostatin L2 crystallizes as a monomer with an 'unusual' conformation of the N-terminus, while sialostatin L crystallizes as a domain-swapped dimer with an N-terminal conformation similar to other cystatins. Deletion of the 'unusual' N-terminal five residues of sialostatin L2 results in marked changes in its selectivity, suggesting that this region is a particularly important determinant of the biochemical activity of sialostatin L2. Collectively, our results reveal the structure of two tick salivary components that facilitate vector blood feeding and that one of them also supports pathogen transmission to the vertebrate host.« less

[Prokaryotic expression systems].

PubMed

Porowińska, Dorota; Wujak, Magdalena; Roszek, Katarzyna; Komoszyński, Michał

2013-03-01

For overproduction of recombinant proteins both eukaryotic and prokaryotic expression systems are used. Choosing the right system depends, among other things, on the growth rate and culture of host cells, level of the target gene expression and posttranslational processing of the synthesized protein. Regardless of the type of expression system, its basic elements are the vector and the expression host. The most widely used system for protein overproduction, both on a laboratory and industrial scale, is the prokaryotic system. This system is based primarily on the bacteria E. coli, although increasingly often Bacillus species are used. The prokaryotic system allows one to obtain large quantities of recombinant proteins in a short time. A simple and inexpensive bacterial cell culture and well-known mechanisms of transcription and translation facilitate the use of these microorganisms. The simplicity of genetic modifications and the availability of many bacterial mutants are additional advantages of the prokaryotic system. In this article we characterize the structural elements of prokaryotic expression vectors. Also strategies for preparation of the target protein gene that increase productivity, facilitate detection and purification of recombinant protein and provide its activity are discussed. Bacterial strains often used as host cells in expression systems as well as the potential location of heterologous proteins are characterized. Knowledge of the basic elements of the prokaryotic expression system allows for production of biologically active proteins in a short time and in satisfactory quantities. 

Transesophageal versus transcranial motor evoked potentials to monitor spinal cord ischemia.

PubMed

Tsuda, Kazumasa; Shiiya, Norihiko; Takahashi, Daisuke; Ohkura, Kazuhiro; Yamashita, Katsushi; Kando, Yumi; Arai, Yoshifumi

2016-02-01

We have previously reported that transesophageal motor evoked potential is feasible and more stable than transcranial motor evoked potential. This study aimed to investigate the efficacy of transesophageal motor evoked potential to monitor spinal cord ischemia. Transesophageal and transcranial motor evoked potentials were recorded in 13 anesthetized dogs at the bilateral forelimbs, anal sphincters, and hindlimbs. Spinal cord ischemia was induced by aortic balloon occlusion at the 8th to 10th thoracic vertebra level. In the 12 animals with motor evoked potential disappearance, occlusion was maintained for 10 minutes (n = 6) or 40 minutes (n = 6) after motor evoked potential disappearance. Neurologic function was evaluated by Tarlov score at 24 and 48 hours postoperatively. Time to disappearance of bilateral motor evoked potentials was quicker in transesophageal motor evoked potentials than in transcranial motor evoked potentials at anal sphincters (6.9 ± 3.1 minutes vs 8.3 ± 3.4 minutes, P = .02) and hindlimbs (5.7 ± 1.9 minutes vs 7.1 ± 2.7 minutes, P = .008). Hindlimb function was normal in all dogs in the 10-minute occlusion group, and motor evoked potentials recovery (>75% on both sides) after reperfusion was quicker in transesophageal motor evoked potentials than transcranial motor evoked potentials at hindlimbs (14.8 ± 5.6 minutes vs 24.7 ± 8.2 minutes, P = .001). At anal sphincters, transesophageal motor evoked potentials always reappeared (>25%), but transcranial motor evoked potentials did not in 3 of 6 dogs. In the 40-minute occlusion group, hindlimb motor evoked potentials did not reappear in 4 dogs with paraplegia. Among the 2 remaining dogs, 1 with paraparesis (Tarlov 3) showed delayed recovery (>75%) of hindlimb motor evoked potentials without reappearance of anal sphincter motor evoked potentials. In another dog with spastic paraplegia, transesophageal motor evoked potentials from the hindlimbs remained less than 20%, whereas transcranial motor evoked potentials showed recovery (>75%). Transesophageal motor evoked potentials may be superior to transcranial motor evoked potentials in terms of quicker response to spinal cord ischemia and better prognostic value. Copyright © 2016 The American Association for Thoracic Surgery. Published by Elsevier Inc. All rights reserved.

Shift schedule, work-family relationships, marital communication, job satisfaction and health among transport service shift workers.

PubMed

Iskra-Golec, Irena; Smith, Lawrence; Wilczek-Rużyczka, Ewa; Siemiginowska, Patrycja; Wątroba, Joanna

2017-02-21

Existing research has documented that shiftwork consequences may depend on the shift system parameters. Fast rotating systems (1-3 shifts of the same kind in a row) and day work have been found to be less disruptive biologically and socially than slower rotating systems and afternoon and night work. The aim of this study was to compare day workers and shift workers of different systems in terms of rotation speed and shifts worked with regard to work-family and family-work positive and negative spillover, marital communication style, job satisfaction and health. Employees (N = 168) of the maintenance workshops of transportation service working different shift systems (day shift, weekly rotating 2 and 3‑shift system, and fast rotating 3-shift system) participated in the study. They completed the Work- Family Spillover Questionnaire, Marital Communication Questionnaire, Minnesota Job Satisfaction Questionnaire and the Physical Health Questionnaire (a part of the Standard Shiftwork Index). The workers of quicker rotating 3-shift systems reported significantly higher scores of family-to-work facilitation (F(3, 165) = 4.175, p = 0.007) and a higher level of constructive style of marital communication (Engagement F(3, 165) = 2.761, p = 0.044) than the workers of slower rotating 2-shift systems. There were no differences between the groups of workers with regard to health and job satisfaction. A higher level of work-family facilitation and a more constructive style of marital communication were found among the workers of faster rotating 3-shift system when compared to the workers of a slower rotating 2-shift system (afternoon, night). This may indicate that the fast rotating shift system in contrary to the slower rotating one is more friendly for the work and family domains and for the relationship between them. Int J Occup Med Environ Health 2017;30(1):121-131. This work is available in Open Access model and licensed under a CC BY-NC 3.0 PL license.

Regional-Scale Chemical Transport Modeling in Support of the Ace-Asia Experiment

NASA Astrophysics Data System (ADS)

Guttikunda, S.; Uno, I.; Carmichael, G.; Streets, D. G.; Tang, Y.; Yienger, J. J.; Woo, J.; Thongboonchoo, N.; Clarke, A.; Blomquist, B.; McNaughton, C.

2002-12-01

Chemical Transport Models (CTMs) are playing increasingly important roles in the design, execution, and analysis of large-scale atmospheric chemistry field studies. They are being used in forecast-mode to enhance flight planning by enabling the representation of important three-dimensional atmospheric chemical structures (such as dust storm plumes, polluted air masses associated with large cities, and widespread biomass burning events) and how they evolve over time. CTM forecasts play the additional important roles of providing a 4-dimensional contextual representation of the experiment, and facilitating a quicker analysis of the field results. CTMs also facilitate the integration of the different measurements and measurement platforms (e.g., aircraft, ground stations and satellite observations). Finally, CTMs play an important role in evaluating and helping to improve emission estimates. We developed the Chemical weather FORecasting System (CFORS) to assist in the analysis of field experiments. CFORS was applied in the design and execution of the TRACE-P, ACE-Asia and the ITCT-Y2K intensive field experiments. In this paper we present a brief overview of the CFORS system, and focus on the use of the CTM on the analysis of the Ace-Asia data. ACE-Asia produced a high density of data from multiple independent measurements, and a wide variety of different types of data. These include: a) in-situ measurements from ground stations, on board ships, and on various aircraft platforms; b) remote sensed data from satellites; and c) vertical distributions from ground based lidars. This data spans vast temporal and spatial scales, and illustrates the needs and challenges of integrating these data into a consistent data analysis set. In this paper we plan to illustrate efforts to integrate measured and modeled data into a comprehensive data set for analysis by measurers and modelers. We will also outline future directions and challenges.

Robot Faces that Follow Gaze Facilitate Attentional Engagement and Increase Their Likeability

PubMed Central

Willemse, Cesco; Marchesi, Serena; Wykowska, Agnieszka

2018-01-01

Gaze behavior of humanoid robots is an efficient mechanism for cueing our spatial orienting, but less is known about the cognitive–affective consequences of robots responding to human directional cues. Here, we examined how the extent to which a humanoid robot (iCub) avatar directed its gaze to the same objects as our participants affected engagement with the robot, subsequent gaze-cueing, and subjective ratings of the robot’s characteristic traits. In a gaze-contingent eyetracking task, participants were asked to indicate a preference for one of two objects with their gaze while an iCub avatar was presented between the object photographs. In one condition, the iCub then shifted its gaze toward the object chosen by a participant in 80% of the trials (joint condition) and in the other condition it looked at the opposite object 80% of the time (disjoint condition). Based on the literature in human–human social cognition, we took the speed with which the participants looked back at the robot as a measure of facilitated reorienting and robot-preference, and found these return saccade onset times to be quicker in the joint condition than in the disjoint condition. As indicated by results from a subsequent gaze-cueing tasks, the gaze-following behavior of the robot had little effect on how our participants responded to gaze cues. Nevertheless, subjective reports suggested that our participants preferred the iCub following participants’ gaze to the one with a disjoint attention behavior, rated it as more human-like and as more likeable. Taken together, our findings show a preference for robots who follow our gaze. Importantly, such subtle differences in gaze behavior are sufficient to influence our perception of humanoid agents, which clearly provides hints about the design of behavioral characteristics of humanoid robots in more naturalistic settings. PMID:29459842

Group-velocity-locked vector soliton molecules in fiber lasers.

PubMed

Luo, Yiyang; Cheng, Jianwei; Liu, Bowen; Sun, Qizhen; Li, Lei; Fu, Songnian; Tang, Dingyuan; Zhao, Luming; Liu, Deming

2017-05-24

Physics phenomena of multi-soliton complexes have enriched the life of dissipative solitons in fiber lasers. By developing a birefringence-enhanced fiber laser, we report the first experimental observation of group-velocity-locked vector soliton (GVLVS) molecules. The birefringence-enhanced fiber laser facilitates the generation of GVLVSs, where the two orthogonally polarized components are coupled together to form a multi-soliton complex. Moreover, the interaction of repulsive and attractive forces between multiple pulses binds the particle-like GVLVSs together in time domain to further form compound multi-soliton complexes, namely GVLVS molecules. By adopting the polarization-resolved measurement, we show that the two orthogonally polarized components of the GVLVS molecules are both soliton molecules supported by the strongly modulated spectral fringes and the double-humped intensity profiles. Additionally, GVLVS molecules with various soliton separations are also observed by adjusting the pump power and the polarization controller.

Next generation tools for high-throughput promoter and expression analysis employing single-copy knock-ins at the Hprt1 locus.

PubMed

Yang, G S; Banks, K G; Bonaguro, R J; Wilson, G; Dreolini, L; de Leeuw, C N; Liu, L; Swanson, D J; Goldowitz, D; Holt, R A; Simpson, E M

2009-03-01

We have engineered a set of useful tools that facilitate targeted single copy knock-in (KI) at the hypoxanthine guanine phosphoribosyl transferase 1 (Hprt1) locus. We employed fine scale mapping to delineate the precise breakpoint location at the Hprt1(b-m3) locus allowing allele specific PCR assays to be established. Our suite of tools contains four targeting expression vectors and a complementing series of embryonic stem cell lines. Two of these vectors encode enhanced green fluorescent protein (EGFP) driven by the human cytomegalovirus immediate-early enhancer/modified chicken beta-actin (CAG) promoter, whereas the other two permit flexible combinations of a chosen promoter combined with a reporter and/or gene of choice. We have validated our tools as part of the Pleiades Promoter Project (http://www.pleiades.org), with the generation of brain-specific EGFP positive germline mouse strains.

Support vector machines and generalisation in HEP

NASA Astrophysics Data System (ADS)

Bevan, Adrian; Gamboa Goñi, Rodrigo; Hays, Jon; Stevenson, Tom

2017-10-01

We review the concept of Support Vector Machines (SVMs) and discuss examples of their use in a number of scenarios. Several SVM implementations have been used in HEP and we exemplify this algorithm using the Toolkit for Multivariate Analysis (TMVA) implementation. We discuss examples relevant to HEP including background suppression for H → τ + τ - at the LHC with several different kernel functions. Performance benchmarking leads to the issue of generalisation of hyper-parameter selection. The avoidance of fine tuning (over training or over fitting) in MVA hyper-parameter optimisation, i.e. the ability to ensure generalised performance of an MVA that is independent of the training, validation and test samples, is of utmost importance. We discuss this issue and compare and contrast performance of hold-out and k-fold cross-validation. We have extended the SVM functionality and introduced tools to facilitate cross validation in TMVA and present results based on these improvements.

Implant Composed of Demineralized Bone and Mesenchymal Stem Cells Genetically Modified with AdBMP2/AdBMP7 for the Regeneration of Bone Fractures in Ovis aries.

PubMed

Hernandez-Hurtado, Adelina A; Borrego-Soto, Gissela; Marino-Martinez, Ivan A; Lara-Arias, Jorge; Romero-Diaz, Viktor J; Abrego-Guerra, Adalberto; Vilchez-Cavazos, Jose F; Elizondo-Riojas, Guillermo; Martinez-Rodriguez, Herminia G; Espinoza-Juarez, Marcela A; Lopez-Romero, Gloria C; Robles-Zamora, Alejandro; Mendoza Lemus, Oscar F; Ortiz-Lopez, Rocio; Rojas-Martinez, Augusto

2016-01-01

Adipose-derived mesenchymal stem cells (ADMSCs) are inducible to an osteogenic phenotype by the bone morphogenetic proteins (BMPs). This facilitates the generation of implants for bone tissue regeneration. This study evaluated the in vitro osteogenic differentiation of ADMSCs transduced individually and in combination with adenoviral vectors expressing BMP2 and BMP7. Moreover, the effectiveness of the implant containing ADMSCs transduced with the adenoviral vectors AdBMP2/AdBMP7 and embedded in demineralized bone matrix (DBM) was tested in a model of tibial fracture in sheep. This graft was compared to ewes implanted with untransduced ADMSCs embedded in the same matrix and with injured but untreated animals. In vivo results showed accelerated osteogenesis in the group treated with the AdBMP2/AdBMP7 transduced ADMSC graft, which also showed improved restoration of the normal bone morphology.

Application of inertial instruments for DSN antenna pointing and tracking

NASA Technical Reports Server (NTRS)

Eldred, D. B.; Nerheim, N. M.; Holmes, K. G.

1990-01-01

The feasibility of using inertial instruments to determine the pointing attitude of the NASA Deep Space Network antennas is examined. The objective is to obtain 1 mdeg pointing knowledge in both blind pointing and tracking modes to facilitate operation of the Deep Space Network 70 m antennas at 32 GHz. A measurement system employing accelerometers, an inclinometer, and optical gyroscopes is proposed. The initial pointing attitude is established by determining the direction of the local gravity vector using the accelerometers and the inclinometer, and the Earth's spin axis using the gyroscopes. Pointing during long-term tracking is maintained by integrating the gyroscope rates and augmenting these measurements with knowledge of the local gravity vector. A minimum-variance estimator is used to combine measurements to obtain the antenna pointing attitude. A key feature of the algorithm is its ability to recalibrate accelerometer parameters during operation. A survey of available inertial instrument technologies is also given.

Immune responses to baculovirus-displayed enterovirus 71 VP1 antigen.

PubMed

Kiener, Tanja K; Premanand, Balraj; Kwang, Jimmy

2013-04-01

The increased distribution and neurovirulence of enterovirus 71 is an important health threat for young children in Asia Pacific. Vaccine design has concentrated on inactivated virus with the most advanced undergoing Phase III clinical trials. By using a subunit vaccine approach, production costs could be reduced by lowering the need for biocontainment. In addition, novel mutations could be rapidly incorporated to reflect the emergence of new enterovirus 71 subgenogroups. To circumvent the problems associated with conventional subunit vaccines, the antigen can be displayed on a viral vector that conveys stability and facilitates purification. Additional advantages of viral-vectored subunit vaccines are their ability to stimulate the innate immune system by transducing cells and the possibility of oral or nasal delivery, which dispenses with the need for syringes and medical personnel. Baculovirus-displayed VP1 combines all these benefits with protection that is as efficient as inactivated virus.

Use of the vector index and geographic information system to prospectively inform West Nile virus interventions.

PubMed

Jones, Roderick C; Weaver, Kingsley N; Smith, Shamika; Blanco, Claudia; Flores, Cristina; Gibbs, Kevin; Markowski, Daniel; Mutebi, John-Paul

2011-09-01

We sought to estimate West Nile virus (WNV) activity in mosquito populations weekly at the census tract level in Chicago, IL, and to provide this information graphically. Each week we calculated a vector index (VI) for each mosquito trap then generated tract estimates using geographic information systems. During June 29-September 13, 2008, a median of 527 (60%) of 874 possible tracts per week had a VI value. Overall, 94% of the weekly VI tract estimates were 0; among those with a VI estimate greater than 0, the median was 0.33 (range 0.003-3.5). Officials deemed risk levels and weather conditions appropriate for adulticide treatments on 3 occasions, resulting in the treatment of approximately 252 linear kilometers of residential streets and alleys. Our analysis successfully converted complex, raw surveillance data into a format that highlighted areas of elevated WNV activity and facilitated the determination of appropriate response procedures.

Design of magnetic polyplexes taken up efficiently by dendritic cell for enhanced DNA vaccine delivery.

PubMed

Nawwab Al-Deen, F M; Selomulya, C; Kong, Y Y; Xiang, S D; Ma, C; Coppel, R L; Plebanski, M

2014-02-01

Dendritic cells (DC) targeting vaccines require high efficiency for uptake, followed by DC activation and maturation. We used magnetic vectors comprising polyethylenimine (PEI)-coated superparamagnetic iron oxide nanoparticles, with hyaluronic acid (HA) of different molecular weights (<10 and 900 kDa) to reduce cytotoxicity and to facilitate endocytosis of particles into DCs via specific surface receptors. DNA encoding Plasmodium yoelii merozoite surface protein 1-19 and a plasmid encoding yellow fluorescent gene were added to the magnetic complexes with various % charge ratios of HA: PEI. The presence of magnetic fields significantly enhanced DC transfection and maturation. Vectors containing a high-molecular-weight HA with 100% charge ratio of HA: PEI yielded a better transfection efficiency than others. This phenomenon was attributed to their longer molecular chains and higher mucoadhesive properties aiding DNA condensation and stability. Insights gained should improve the design of more effective DNA vaccine delivery systems.

Integrated vector management: the Zambian experience.

PubMed

Chanda, Emmanuel; Masaninga, Fred; Coleman, Michael; Sikaala, Chadwick; Katebe, Cecilia; Macdonald, Michael; Baboo, Kumar S; Govere, John; Manga, Lucien

2008-08-27

The Zambian Malaria Control Programme with the Roll Back Malaria (RBM) partners have developed the current National Malaria Strategic Plan (NMSP 2006-2011) which focuses on prevention based on the Integrated Vector Management (IVM) strategy. The introduction and implementation of an IVM strategy was planned in accordance with the World Health Organization (WHO) steps towards IVM implementation namely Introduction Phase, Consolidation Phase and Expansion Phase. IVM has created commitment for Legal and Regulatory policy review, monitoring, Research and a strong stewardship by the chemical suppliers. It has also leveraged additional resources, improved inter-sectoral collaboration, capacity building and enhanced community participation which facilitated a steady scaling up in coverage and utilisation of key preventive interventions. Thus, markedly reducing malaria incidence and case fatalities in the country. Zambia has successfully introduced, consolidated and expanded IVM activities. Resulting in increased coverage and utilization of interventions and markedly reducing malaria-related morbidity and mortality while ensuring a better protection of the environment.

Correcting systematic errors in high-sensitivity deuteron polarization measurements

NASA Astrophysics Data System (ADS)

Brantjes, N. P. M.; Dzordzhadze, V.; Gebel, R.; Gonnella, F.; Gray, F. E.; van der Hoek, D. J.; Imig, A.; Kruithof, W. L.; Lazarus, D. M.; Lehrach, A.; Lorentz, B.; Messi, R.; Moricciani, D.; Morse, W. M.; Noid, G. A.; Onderwater, C. J. G.; Özben, C. S.; Prasuhn, D.; Levi Sandri, P.; Semertzidis, Y. K.; da Silva e Silva, M.; Stephenson, E. J.; Stockhorst, H.; Venanzoni, G.; Versolato, O. O.

2012-02-01

This paper reports deuteron vector and tensor beam polarization measurements taken to investigate the systematic variations due to geometric beam misalignments and high data rates. The experiments used the In-Beam Polarimeter at the KVI-Groningen and the EDDA detector at the Cooler Synchrotron COSY at Jülich. By measuring with very high statistical precision, the contributions that are second-order in the systematic errors become apparent. By calibrating the sensitivity of the polarimeter to such errors, it becomes possible to obtain information from the raw count rate values on the size of the errors and to use this information to correct the polarization measurements. During the experiment, it was possible to demonstrate that corrections were satisfactory at the level of 10 -5 for deliberately large errors. This may facilitate the real time observation of vector polarization changes smaller than 10 -6 in a search for an electric dipole moment using a storage ring.

Self-focusing therapeutic gene delivery with intelligent gene vector swarms: intra-swarm signalling through receptor transgene expression in targeted cells.

PubMed

Tolmachov, Oleg E

2015-01-01

Gene delivery in vivo that is tightly focused on the intended target cells is essential to maximize the benefits of gene therapy and to reduce unwanted side-effects. Cell surface markers are immediately available for probing by therapeutic gene vectors and are often used to direct gene transfer with these vectors to specific target cell populations. However, it is not unusual for the choice of available extra-cellular markers to be too scarce to provide a reliable definition of the desired therapeutically relevant set of target cells. Therefore, interrogation of intra-cellular determinants of cell-specificity, such as tissue-specific transcription factors, can be vital in order to provide detailed cell-guiding information to gene vector particles. An important improvement in cell-specific gene delivery can be achieved through auto-buildup in vector homing efficiency using intelligent 'self-focusing' of swarms of vector particles on target cells. Vector self-focusing was previously suggested to rely on the release of diffusible chemo-attractants after a successful target-specific hit by 'scout' vector particles. I hypothesize that intelligent self-focusing behaviour of swarms of cell-targeted therapeutic gene vectors can be accomplished without the employment of difficult-to-use diffusible chemo-attractants, instead relying on the intra-swarm signalling through cells expressing a non-diffusible extra-cellular receptor for the gene vectors. In the proposed model, cell-guiding information is gathered by the 'scout' gene vector particles, which: (1) attach to a variety of cells via a weakly binding (low affinity) receptor; (2) successfully facilitate gene transfer into these cells; (3) query intra-cellular determinants of cell-specificity with their transgene expression control elements and (4) direct the cell-specific biosynthesis of a vector-encoded strongly binding (high affinity) cell-surface receptor. Free members of the vector swarm loaded with therapeutic cargo are then attracted to and internalized into the intended target cells via the expressed cognate strongly binding extra-cellular receptor, causing escalation of gene transfer into these cells and increasing the copy number of the therapeutic gene expression modules. Such self-focusing swarms of gene vectors can be either homogeneous, with 'scout' and 'therapeutic' members of the swarm being structurally identical, or, alternatively, heterogeneous (split), with 'scout' and 'therapeutic' members of the swarm being structurally specialized. It is hoped that the proposed self-focusing cell-targeted gene vector swarms with receptor-mediated intra-swarm signalling could be particularly effective in 'top-up' gene delivery scenarios, achieving high-level and sustained expression of therapeutic transgenes that are prone to shut-down through degradation and silencing. Crucially, in contrast to low-precision 'general location' vector guidance by diffusible chemo-attractants, ear-marking non-diffusible receptors can provide high-accuracy targeting of therapeutic vector particles to the specific cell, which has undergone a 'successful cell-specific hit' by a 'scout' vector particle. Opportunities for cell targeting could be expanded, since in the proposed model of self-focusing it could be possible to probe a broad selection of intra-cellular determinants of cell-specificity and not just to rely exclusively on extra-cellular markers of cell-specificity. By employing such self-focusing gene vectors for the improvement of cell-targeted delivery of therapeutic genes, e.g., in cancer therapy or gene addition therapy of recessive genetic diseases, it could be possible to broaden a leeway for the reduction of the vector load and, consequently, to minimize undesired vector cytotoxicity, immune reactions, and the risk of inadvertent genetic modification of germline cells in genetic treatment in vivo. Copyright © 2014 Elsevier B.V. All rights reserved.

Dengue virus type 2: replication and tropisms in orally infected Aedes aegypti mosquitoes.

PubMed

Salazar, Ma Isabel; Richardson, Jason H; Sánchez-Vargas, Irma; Olson, Ken E; Beaty, Barry J

2007-01-30

To be transmitted by its mosquito vector, dengue virus (DENV) must infect midgut epithelial cells, replicate and disseminate into the hemocoel, and finally infect the salivary glands, which is essential for transmission. The extrinsic incubation period (EIP) is very relevant epidemiologically and is the time required from the ingestion of virus until it can be transmitted to the next vertebrate host. The EIP is conditioned by the kinetics and tropisms of virus replication in its vector. Here we document the virogenesis of DENV-2 in newly-colonized Aedes aegypti mosquitoes from Chetumal, Mexico in order to understand better the effect of vector-virus interactions on dengue transmission. After ingestion of DENV-2, midgut infections in Chetumal mosquitoes were characterized by a peak in virus titers between 7 and 10 days post-infection (dpi). The amount of viral antigen and viral titers in the midgut then declined, but viral RNA levels remained stable. The presence of DENV-2 antigen in the trachea was positively correlated with virus dissemination from the midgut. DENV-2 antigen was found in salivary gland tissue in more than a third of mosquitoes at 4 dpi. Unlike in the midgut, the amount of viral antigen (as well as the percent of infected salivary glands) increased with time. DENV-2 antigen also accumulated and increased in neural tissue throughout the EIP. DENV-2 antigen was detected in multiple tissues of the vector, but unlike some other arboviruses, was not detected in muscle. Our results suggest that the EIP of DENV-2 in its vector may be shorter that the previously reported and that the tracheal system may facilitate DENV-2 dissemination from the midgut. Mosquito organs (e.g. midgut, neural tissue, and salivary glands) differed in their response to DENV-2 infection.

Potential distribution of mosquito vector species in a primary malaria endemic region of Colombia

PubMed Central

Altamiranda-Saavedra, Mariano; Arboleda, Sair; Parra, Juan L.; Peterson, A. Townsend

2017-01-01

Rapid transformation of natural ecosystems changes ecological conditions for important human disease vector species; therefore, an essential task is to identify and understand the variables that shape distributions of these species to optimize efforts toward control and mitigation. Ecological niche modeling was used to estimate the potential distribution and to assess hypotheses of niche similarity among the three main malaria vector species in northern Colombia: Anopheles nuneztovari, An. albimanus, and An. darlingi. Georeferenced point collection data and remotely sensed, fine-resolution satellite imagery were integrated across the Urabá –Bajo Cauca–Alto Sinú malaria endemic area using a maximum entropy algorithm. Results showed that An. nuneztovari has the widest geographic distribution, occupying almost the entire study region; this niche breadth is probably related to the ability of this species to colonize both, natural and disturbed environments. The model for An. darlingi showed that most suitable localities for this species in Bajo Cauca were along the Cauca and Nechí river. The riparian ecosystems in this region and the potential for rapid adaptation by this species to novel environments, may favor the establishment of populations of this species. Apparently, the three main Colombian Anopheles vector species in this endemic area do not occupy environments either with high seasonality, or with low seasonality and high NDVI values. Estimated overlap in geographic space between An. nuneztovari and An. albimanus indicated broad spatial and environmental similarity between these species. An. nuneztovari has a broader niche and potential distribution. Dispersal ability of these species and their ability to occupy diverse environmental situations may facilitate sympatry across many environmental and geographic contexts. These model results may be useful for the design and implementation of malaria species-specific vector control interventions optimized for this important malaria region. PMID:28594942

Gene trapping in differentiating cell lines: regulation of the lysosomal protease cathepsin B in skeletal myoblast growth and fusion.

PubMed

Gogos, J A; Thompson, R; Lowry, W; Sloane, B F; Weintraub, H; Horwitz, M

1996-08-01

To identify genes regulated during skeletal muscle differentiation, we have infected mouse C2C12 myoblasts with retroviral gene trap vectors, containing a promoterless marker gene with a 5' splice acceptor signal. Integration of the vector adjacent to an actively transcribed gene places the marker under the transcriptional control of the endogenous gene, while the adjacent vector sequences facilitate cloning. The vector insertionally mutates the trapped locus and may also form fusion proteins with the endogenous gene product. We have screened several hundred clones, each containing a trapping vector integrated into a different endogenous gene. In agreement with previous estimates based on hybridization kinetics, we find that a large proportion of all genes expressed in myoblasts are regulated during differentiation. Many of these genes undergo unique temporal patterns of activation or repression during cell growth and myotube formation, and some show specific patterns of subcellular localization. The first gene we have identified with this strategy is the lysosomal cysteine protease cathepsin B. Expression from the trapped allele is upregulated during early myoblast fusion and downregulated in myotubes. A direct role for cathepsin B in myoblast growth and fusion is suggested by the observation that the trapped cells deficient in cathepsin B activity have an unusual morphology and reduced survival in low-serum media and undergo differentiation with impaired cellular fusion. The phenotype is reproduced by antisense cathepsin B expression in parental C2C12 myoblasts. The cellular phenotype is similar to that observed in cultured myoblasts from patients with I cell disease, in which there is diminished accumulation of lysosomal enzymes. This suggests that a specific deficiency of cathepsin B could contribute to the myopathic component of this illness.

Towards a resource-based habitat approach for spatial modelling of vector-borne disease risks.

PubMed

Hartemink, Nienke; Vanwambeke, Sophie O; Purse, Bethan V; Gilbert, Marius; Van Dyck, Hans

2015-11-01

Given the veterinary and public health impact of vector-borne diseases, there is a clear need to assess the suitability of landscapes for the emergence and spread of these diseases. Current approaches for predicting disease risks neglect key features of the landscape as components of the functional habitat of vectors or hosts, and hence of the pathogen. Empirical-statistical methods do not explicitly incorporate biological mechanisms, whereas current mechanistic models are rarely spatially explicit; both methods ignore the way animals use the landscape (i.e. movement ecology). We argue that applying a functional concept for habitat, i.e. the resource-based habitat concept (RBHC), can solve these issues. The RBHC offers a framework to identify systematically the different ecological resources that are necessary for the completion of the transmission cycle and to relate these resources to (combinations of) landscape features and other environmental factors. The potential of the RBHC as a framework for identifying suitable habitats for vector-borne pathogens is explored and illustrated with the case of bluetongue virus, a midge-transmitted virus affecting ruminants. The concept facilitates the study of functional habitats of the interacting species (vectors as well as hosts) and provides new insight into spatial and temporal variation in transmission opportunities and exposure that ultimately determine disease risks. It may help to identify knowledge gaps and control options arising from changes in the spatial configuration of key resources across the landscape. The RBHC framework may act as a bridge between existing mechanistic and statistical modelling approaches. © 2014 The Authors. Biological Reviews published by John Wiley & Sons Ltd on behalf of Cambridge Philosophical Society.

Floral Scent Mimicry and Vector-Pathogen Associations in a Pseudoflower-Inducing Plant Pathogen System

PubMed Central

McArt, Scott H.; Miles, Timothy D.; Rodriguez-Saona, Cesar; Schilder, Annemiek; Adler, Lynn S.; Grieshop, Matthew J.

2016-01-01

Several fungal plant pathogens induce ‘pseudoflowers’ on their hosts to facilitate insect-mediated transmission of gametes and spores. When spores must be transmitted to host flowers to complete the fungal life cycle, we predict that pseudoflowers should evolve traits that mimic flowers and attract the most effective vectors in the flower-visiting community. We quantified insect visitation to flowers, healthy leaves and leaves infected with Monilinia vaccinii-corymbosi (Mvc), the causative agent of mummy berry disease of blueberry. We developed a nested PCR assay for detecting Mvc spores on bees, flies and other potential insect vectors. We also collected volatiles from blueberry flowers, healthy leaves and leaves infected with Mvc, and experimentally manipulated specific pathogen-induced volatiles to assess attractiveness to potential vectors. Bees and flies accounted for the majority of contacts with flowers, leaves infected with Mvc and healthy leaves. Flowers were contacted most often, while there was no difference between bee or fly contacts with healthy and infected leaves. While bees contacted flowers more often than flies, flies contacted infected leaves more often than bees. Bees were more likely to have Mvc spores on their bodies than flies, suggesting that bees may be more effective vectors than flies for transmitting Mvc spores to flowers. Leaves infected with Mvc had volatile profiles distinct from healthy leaves but similar to flowers. Two volatiles produced by flowers and infected leaves, cinnamyl alcohol and cinnamic aldehyde, were attractive to bees, while no volatiles manipulated were attractive to flies or any other insects. These results suggest that Mvc infection of leaves induces mimicry of floral volatiles, and that transmission occurs primarily via bees, which had the highest likelihood of carrying Mvc spores and visited flowers most frequently. PMID:27851747

Hydrologic variability and the dynamics of West Nile virus transmission

NASA Astrophysics Data System (ADS)

Shaman, J. L.

2011-12-01

West Nile virus (WNV) first emerged in North America in New York City during 1999 and since that time has spread throughout the continent and settled into a pattern of local endemicity in which outbreaks of variable size develop in some years but not others. Predicting where and when these outbreaks will develop is an issue of considerable public health importance. Spillover transmission of WNV to humans typically occurs when infection rates among vector mosquitoes are elevated. Mosquito infection rates are not constant through time but instead increase when newly emergent mosquitoes can more readily acquire WNV by blood-meal feeding on available, infected animal hosts. Such an increase of vector mosquito infection rates is termed amplification and is facilitated for WNV by intense zoonotic transmission of the virus among vector mosquitoes and avian hosts. Theory, observation and model simulations indicate that amplification is favored when mosquito breeding habitats and bird nesting and roosting habitats overlap. Both vector mosquitoes and vertebrate hosts depend on water resources; mosquitoes are critically dependent on the availability of standing water, as the first 3 stages of the mosquito life cycle, egg, larvae, pupae, are aquatic. Here it is shown that hydrologic variability often determines where and when vector mosquitoes and avian hosts congregate together, and when the amplification of WNV is more likely. Measures of land surface wetness and pooling, from ground observation, satellite observation, or numerical modeling, can provide reliable estimates of where and when WNV transmission hotspots will arise. Examples of this linkage between hydrology and WNV activity are given for Florida, Colorado and New York, and an operational system for monitoring and forecasting WNV risk in space and time is presented for Florida.

Highly efficient gene transfer in naive human T cells with a murine leukemia virus-based vector.

PubMed

Dardalhon, V; Jaleco, S; Rebouissou, C; Ferrand, C; Skander, N; Swainson, L; Tiberghien, P; Spits, H; Noraz, N; Taylor, N

2000-08-01

Retroviral vectors based on the Moloney murine leukemia virus (MuLV) have become the primary tool for gene delivery into hematopoietic cells, but clinical trials have been hampered by low transduction efficiencies. Recently, we and others have shown that gene transfer of MuLV-based vectors into T cells can be significantly augmented using a fibronectin-facilitated protocol. Nevertheless, the relative abilities of naive (CD45RA(+)) and memory (CD45RO(+)) lymphocyte subsets to be transduced has not been assessed. Although naive T cells demonstrate a restricted cytokine profile following antigen stimulation and a decreased susceptibility to infection with human immunodeficiency virus, it was not clear whether they could be efficiently infected with a MuLV vector. This study describes conditions that permitted gene transfer of an enhanced green fluorescent protein-expressing retroviral vector in more than 50% of naive umbilical cord (UC) blood and peripheral blood (PB) T cells following CD3/CD28 ligation. Moreover, treatment of naive T cells with interleukin-7 resulted in the maintenance of a CD45RA phenotype and gene transfer levels approached 20%. Finally, it was determined that parameters for optimal transduction of CD45RA(+) T cells isolated from PB and UC blood differed: transduction of the UC cells was significantly increased by the presence of autologous mononuclear cells (24.5% versus 56.5%). Because naive T cells harbor a receptor repertoire that allows them to respond to novel antigens, the development of protocols targeting their transduction is crucial for gene therapy applications. This approach will also allow the functions of exogenous genes to be evaluated in primary nontransformed naive T cells.

Floral Scent Mimicry and Vector-Pathogen Associations in a Pseudoflower-Inducing Plant Pathogen System.

PubMed

McArt, Scott H; Miles, Timothy D; Rodriguez-Saona, Cesar; Schilder, Annemiek; Adler, Lynn S; Grieshop, Matthew J

2016-01-01

Several fungal plant pathogens induce 'pseudoflowers' on their hosts to facilitate insect-mediated transmission of gametes and spores. When spores must be transmitted to host flowers to complete the fungal life cycle, we predict that pseudoflowers should evolve traits that mimic flowers and attract the most effective vectors in the flower-visiting community. We quantified insect visitation to flowers, healthy leaves and leaves infected with Monilinia vaccinii-corymbosi (Mvc), the causative agent of mummy berry disease of blueberry. We developed a nested PCR assay for detecting Mvc spores on bees, flies and other potential insect vectors. We also collected volatiles from blueberry flowers, healthy leaves and leaves infected with Mvc, and experimentally manipulated specific pathogen-induced volatiles to assess attractiveness to potential vectors. Bees and flies accounted for the majority of contacts with flowers, leaves infected with Mvc and healthy leaves. Flowers were contacted most often, while there was no difference between bee or fly contacts with healthy and infected leaves. While bees contacted flowers more often than flies, flies contacted infected leaves more often than bees. Bees were more likely to have Mvc spores on their bodies than flies, suggesting that bees may be more effective vectors than flies for transmitting Mvc spores to flowers. Leaves infected with Mvc had volatile profiles distinct from healthy leaves but similar to flowers. Two volatiles produced by flowers and infected leaves, cinnamyl alcohol and cinnamic aldehyde, were attractive to bees, while no volatiles manipulated were attractive to flies or any other insects. These results suggest that Mvc infection of leaves induces mimicry of floral volatiles, and that transmission occurs primarily via bees, which had the highest likelihood of carrying Mvc spores and visited flowers most frequently.

A stable shuttle vector system for efficient genetic complementation of Helicobacter pylori strains by transformation and conjugation.

PubMed

Heuermann, D; Haas, R

1998-03-01

A versatile plasmid shuttle vector system was constructed, which is useful for genetic complementation of Helicobacter pylori strains or mutants with cloned genes of homologous or heterologous origin. The individual plasmid vectors consist of the minimal essential genetic elements, including an origin of replication for Escherichia coli, a H. pylori-specific replicon originally identified on a small cryptic H. pylori plasmid, an oriT sequence and a multiple cloning site. Shuttle plasmid pHel2 carries a chloramphenicol resistance cassette (catGC) and pHel3 contains a kanamycin resistance gene (aphA-3) as the selectable marker; both are functional in E. coli and H. pylori. The shuttle plasmids were introduced into the H. pylori strain P1 by natural transformation. A efficiency of 7.0 x 10(-7) and 4.7 x 10(-7) transformants per viable recipient was achieved with pHel2 and pHel3, respectively, and both vectors showed stable, autonomous replication in H. pylori. An approximately 100-fold higher H. pylori transformation rate was obtained when the shuttle vectors for transformation were isolated from the homologous H. pylori strain, rather than E. coli, indicating that DNA restriction and modification mechanisms play a crucial role in plasmid transformation. Interestingly, both shuttle vectors could also be mobilized efficiently from E. coli into different H. pylori recipients, with pHel2 showing an efficiency of 2.0 x 10(-5) transconjugants per viable H. pylori P1 recipient. Thus, DNA restriction seems to be strongly reduced or absent during conjugal transfer. The functional complementation of a recA-deficient H. pylori mutant by the cloned H. pylori recA+ gene, and the expression of the heterologous green fluorescent protein (GFP) in H. pylori demonstrate the general usefulness of this system, which will significantly facilitate the molecular analysis of H. pylori virulence factors in the future.

Parasite killing in malaria non-vector mosquito Anopheles culicifacies species B: implication of nitric oxide synthase upregulation.

PubMed

Vijay, Sonam; Rawat, Manmeet; Adak, Tridibes; Dixit, Rajnikant; Nanda, Nutan; Srivastava, Harish; Sharma, Joginder K; Prasad, Godavarthi B K S; Sharma, Arun

2011-04-04

Anopheles culicifacies, the main vector of human malaria in rural India, is a complex of five sibling species. Despite being phylogenetically related, a naturally selected subgroup species B of this sibling species complex is found to be a poor vector of malaria. We have attempted to understand the differences between vector and non-vector Anopheles culicifacies mosquitoes in terms of transcriptionally activated nitric oxide synthase (AcNOS) physiologies to elucidate the mechanism of refractoriness. Identification of the differences between genes and gene products that may impart refractory phenotype can facilitate development of novel malaria transmission blocking strategies. We conducted a study on phylogenetically related susceptible (species A) and refractory (species B) sibling species of An. culicifacies mosquitoes to characterize biochemical and molecular differences in AcNOS gene and gene elements and their ability to inhibit oocyst growth. We demonstrate that in species B, AcNOS specific activity and nitrite/nitrates in mid-guts and haemolymph were higher as compared to species A after invasion of the mid-gut by P. vivax at the beginning and during the course of blood feeding. Semiquantitative RT-PCR and real time PCR data of AcNOS concluded that this gene is more abundantly expressed in midgut of species B than in species A and is transcriptionally upregulated post blood meals. Dietary feeding of L-NAME along with blood meals significantly inhibited midgut AcNOS activity leading to an increase in oocyst production in An. culicifacies species B. We hypothesize that upregulation of mosquito innate cytotoxicity due to NOS in refractory strain to Plasmodium vivax infection may contribute to natural refractoriness in An. culicifacies mosquito population. This innate capacity of refractory mosquitoes could represent the ancestral function of the mosquito immune system against the parasite and could be utilized to understand the molecular basis of refractoriness in planning effective vector control strategies.

Parasite Killing in Malaria Non-Vector Mosquito Anopheles culicifacies Species B: Implication of Nitric Oxide Synthase Upregulation

PubMed Central

Vijay, Sonam; Rawat, Manmeet; Adak, Tridibes; Dixit, Rajnikant; Nanda, Nutan; Srivastava, Harish; Sharma, Joginder K.; Prasad, Godavarthi B. K. S.; Sharma, Arun

2011-01-01

Background Anopheles culicifacies, the main vector of human malaria in rural India, is a complex of five sibling species. Despite being phylogenetically related, a naturally selected subgroup species B of this sibling species complex is found to be a poor vector of malaria. We have attempted to understand the differences between vector and non-vector Anopheles culicifacies mosquitoes in terms of transcriptionally activated nitric oxide synthase (AcNOS) physiologies to elucidate the mechanism of refractoriness. Identification of the differences between genes and gene products that may impart refractory phenotype can facilitate development of novel malaria transmission blocking strategies. Methodology/Principal Findings We conducted a study on phylogenetically related susceptible (species A) and refractory (species B) sibling species of An. culicifacies mosquitoes to characterize biochemical and molecular differences in AcNOS gene and gene elements and their ability to inhibit oocyst growth. We demonstrate that in species B, AcNOS specific activity and nitrite/nitrates in mid-guts and haemolymph were higher as compared to species A after invasion of the mid-gut by P. vivax at the beginning and during the course of blood feeding. Semiquantitative RT-PCR and real time PCR data of AcNOS concluded that this gene is more abundantly expressed in midgut of species B than in species A and is transcriptionally upregulated post blood meals. Dietary feeding of L-NAME along with blood meals significantly inhibited midgut AcNOS activity leading to an increase in oocyst production in An. culicifacies species B. Conclusions/Significance We hypothesize that upregulation of mosquito innate cytotoxicity due to NOS in refractory strain to Plasmodium vivax infection may contribute to natural refractoriness in An. culicifacies mosquito population. This innate capacity of refractory mosquitoes could represent the ancestral function of the mosquito immune system against the parasite and could be utilized to understand the molecular basis of refractoriness in planning effective vector control strategies. PMID:21483693

Annotated Differentially Expressed Salivary Proteins of Susceptible and Insecticide-Resistant Mosquitoes of Anopheles stephensi

PubMed Central

Vijay, Sonam; Rawal, Ritu; Kadian, Kavita; Raghavendra, Kamaraju; Sharma, Arun

2015-01-01

Vector control is one of the major global strategies for control of malaria. However, the major obstacle for vector control is the development of multiple resistances to organochlorine, organophosphorus insecticides and pyrethroids that are currently being used in public health for spraying and in bednets. Salivary glands of vectors are the first target organ for human-vector contact during biting and parasite-vector contact prior to parasite development in the mosquito midguts. The salivary glands secrete anti-haemostatic, anti-inflammatory biologically active molecules to facilitate blood feeding from the host and also inadvertently inject malaria parasites into the vertebrate host. The Anopheles stephensi mosquito, an urban vector of malaria to both human and rodent species has been identified as a reference laboratory model to study mosquito—parasite interactions. In this study, we adopted a conventional proteomic approach of 2D-electrophoresis coupled with MALDI-TOF mass spectrometry and bioinformatics to identify putative differentially expressed annotated functional salivary proteins between An. stephensi susceptible and multiresistant strains with same genetic background. Our results show 2D gel profile and MALDI-TOF comparisons that identified 31 differentially expressed putative modulated proteins in deltamethrin/DDT resistant strains of An. stephensi. Among these 15 proteins were found to be upregulated and 16 proteins were downregulated. Our studies interpret that An. stephensi (multiresistant) caused an upregulated expression of proteins and enzymes like cytochrome 450, short chain dehyrdogenase reductase, phosphodiesterase etc that may have an impact in insecticide resistance and xenobiotic detoxification. Our study elucidates a proteomic response of salivary glands differentially regulated proteins in response to insecticide resistance development which include structural, redox and regulatory enzymes of several pathways. These identified proteins may play a role in regulating mosquito biting behavior patterns and may have implications in the development of malaria parasites in resistant mosquitoes during parasite transmission. PMID:25742511

Annotated differentially expressed salivary proteins of susceptible and insecticide-resistant mosquitoes of Anopheles stephensi.

PubMed

Vijay, Sonam; Rawal, Ritu; Kadian, Kavita; Raghavendra, Kamaraju; Sharma, Arun

2015-01-01

Vector control is one of the major global strategies for control of malaria. However, the major obstacle for vector control is the development of multiple resistances to organochlorine, organophosphorus insecticides and pyrethroids that are currently being used in public health for spraying and in bednets. Salivary glands of vectors are the first target organ for human-vector contact during biting and parasite-vector contact prior to parasite development in the mosquito midguts. The salivary glands secrete anti-haemostatic, anti-inflammatory biologically active molecules to facilitate blood feeding from the host and also inadvertently inject malaria parasites into the vertebrate host. The Anopheles stephensi mosquito, an urban vector of malaria to both human and rodent species has been identified as a reference laboratory model to study mosquito-parasite interactions. In this study, we adopted a conventional proteomic approach of 2D-electrophoresis coupled with MALDI-TOF mass spectrometry and bioinformatics to identify putative differentially expressed annotated functional salivary proteins between An. stephensi susceptible and multiresistant strains with same genetic background. Our results show 2D gel profile and MALDI-TOF comparisons that identified 31 differentially expressed putative modulated proteins in deltamethrin/DDT resistant strains of An. stephensi. Among these 15 proteins were found to be upregulated and 16 proteins were downregulated. Our studies interpret that An. stephensi (multiresistant) caused an upregulated expression of proteins and enzymes like cytochrome 450, short chain dehyrdogenase reductase, phosphodiesterase etc that may have an impact in insecticide resistance and xenobiotic detoxification. Our study elucidates a proteomic response of salivary glands differentially regulated proteins in response to insecticide resistance development which include structural, redox and regulatory enzymes of several pathways. These identified proteins may play a role in regulating mosquito biting behavior patterns and may have implications in the development of malaria parasites in resistant mosquitoes during parasite transmission.

Estimating dengue vector abundance in the wet and dry season: implications for targeted vector control in urban and peri-urban Asia

PubMed Central

Wai, Khin Thet; Arunachalam, Natarajan; Tana, Susilowati; Espino, Fe; Kittayapong, Pattamaporn; Abeyewickreme, W; Hapangama, Dilini; Tyagi, Brij Kishore; Htun, Pe Than; Koyadun, Surachart; Kroeger, Axel; Sommerfeld, Johannes; Petzold, Max

2012-01-01

Background Research has shown that the classical Stegomyia indices (or “larval indices”) of the dengue vector Aedes aegypti reflect the absence or presence of the vector but do not provide accurate measures of adult mosquito density. In contrast, pupal indices as collected in pupal productivity surveys are a much better proxy indicator for adult vector abundance. However, it is unknown when it is most optimal to conduct pupal productivity surveys, in the wet or in the dry season or in both, to inform control services about the most productive water container types and if this pattern varies among different ecological settings. Methods A multi-country study in randomly selected twelve to twenty urban and peri-urban neighborhoods (“clusters”) of six Asian countries, in which all water holding containers were examined for larvae and pupae of Aedes aegypti during the dry season and the wet season and their productivity was characterized by water container types. In addition, meteorological data and information on reported dengue cases were collected. Findings The study reconfirmed the association between rainfall and dengue cases (“dengue season”) and underlined the importance of determining through pupal productivity surveys the “most productive containers types”, responsible for the majority (>70%) of adult dengue vectors. The variety of productive container types was greater during the wet than during the dry season, but included practically all container types productive in the dry season. Container types producing pupae were usually different from those infested by larvae indicating that containers with larval infestations do not necessarily foster pupal development and thus the production of adult Aedes mosquitoes. Conclusion Pupal productivity surveys conducted during the wet season will identify almost all of the most productive container types for both the dry and wet seasons and will therefore facilitate cost-effective targeted interventions. PMID:23318235

A simple method of equine limb force vector analysis and its potential applications.

PubMed

Hobbs, Sarah Jane; Robinson, Mark A; Clayton, Hilary M

2018-01-01

Ground reaction forces (GRF) measured during equine gait analysis are typically evaluated by analyzing discrete values obtained from continuous force-time data for the vertical, longitudinal and transverse GRF components. This paper describes a simple, temporo-spatial method of displaying and analyzing sagittal plane GRF vectors. In addition, the application of statistical parametric mapping (SPM) is introduced to analyse differences between contra-lateral fore and hindlimb force-time curves throughout the stance phase. The overall aim of the study was to demonstrate alternative methods of evaluating functional (a)symmetry within horses. GRF and kinematic data were collected from 10 horses trotting over a series of four force plates (120 Hz). The kinematic data were used to determine clean hoof contacts. The stance phase of each hoof was determined using a 50 N threshold. Vertical and longitudinal GRF for each stance phase were plotted both as force-time curves and as force vector diagrams in which vectors originating at the centre of pressure on the force plate were drawn at intervals of 8.3 ms for the duration of stance. Visual evaluation was facilitated by overlay of the vector diagrams for different limbs. Summary vectors representing the magnitude (VecMag) and direction (VecAng) of the mean force over the entire stance phase were superimposed on the force vector diagram. Typical measurements extracted from the force-time curves (peak forces, impulses) were compared with VecMag and VecAng using partial correlation (controlling for speed). Paired samples t -tests (left v. right diagonal pair comparison and high v. low vertical force diagonal pair comparison) were performed on discrete and vector variables using traditional methods and Hotelling's T 2 tests on normalized stance phase data using SPM. Evidence from traditional statistical tests suggested that VecMag is more influenced by the vertical force and impulse, whereas VecAng is more influenced by the longitudinal force and impulse. When used to evaluate mean data from the group of ten sound horses, SPM did not identify differences between the left and right contralateral limb pairs or between limb pairs classified according to directional asymmetry. When evaluating a single horse, three periods were identified during which differences in the forces between the left and right forelimbs exceeded the critical threshold ( p  < .01). Traditional statistical analysis of 2D GRF peak values, summary vector variables and visual evaluation of force vector diagrams gave harmonious results and both methods identified the same inter-limb asymmetries. As alpha was more tightly controlled using SPM, significance was only found in the individual horse although T 2 plots followed the same trends as discrete analysis for the group. The techniques of force vector analysis and SPM hold promise for investigations of sidedness and asymmetry in horses.

Seismic transmission operator reciprocity - II: impedance-operator symmetry via elastic lateral modes

NASA Astrophysics Data System (ADS)

Thomson, C. J.

2015-08-01

The properties of the overburden transmission response are of particular interest for the analysis of reflectivity illumination or blurring in seismic depth imaging. The first step to showing a transmission-operator reciprocity property is to identify the symmetry of the so-called displacement-to-traction operators. The latter are analogous to Dirichlet-to-Neumann operators and they may also be called impedance operators. Their symmetry is deduced here after development of a formal spectral or modal theory of lateral wavefunctions in a laterally heterogeneous generally anisotropic elastic medium. The elastic lateral modes are displacement-traction 6-vectors and they are built from two auxiliary 3-vector lateral-mode bases. These auxiliary modes arise from Hermitian and anti-Hermitian operators, so they have familiar properties such as orthogonality. There is no assumption of down/up symmetry of the elasticity tensor, but basic assumptions are made about the existence and completeness of the elastic modes. A point-symmetry property appears and plays a central role. The 6-vector elastic modes have a symplectic orthogonality property, which facilitates the development of modal expansions for 6-vector functions of the lateral coordinates when completeness is assumed. While the elastic modal theory is consistent with the laterally homogeneous case, numerical work would provide confidence that it is correct in general. An appendix contains an introductory overview of acoustic lateral modes that were studied by other authors, given from the perspective of this new work. A distinction is drawn between unit normalization of scalar auxiliary modes and a separate energy-flux normalization of 2-vector acoustic modes. Neither is crucial to the form of acoustic pressure-to-velocity or impedance operators. This statement carries over to the elastic case for the 3-vector auxiliary- and 6-vector elastic-mode normalizations. The modal theory is used to construct the kernel of the elastic displacement-to-traction or impedance operator. Symmetry properties of this operator are then deduced, which is the main goal of this paper. The implications of elastic impedance-operator symmetry and the symplectic property for the transmission and reflection responses of finite regions are described in a companion paper.

Triatominae-Trypanosoma cruzi/T. rangeli: Vector-parasite interactions.

PubMed

Vallejo, G A; Guhl, F; Schaub, G A

2009-01-01

Of the currently known 140 species in the family Reduviidae, subfamily Triatominae, those which are most important as vectors of the aetiologic agent of Chagas disease, Trypanosoma cruzi, belong to the tribes Triatomini and Rhodniini. The latter not only transmit T. cruzi but also Trypanosoma rangeli, which is considered apathogenic for the mammalian host but can be pathogenic for the vectors. Using different molecular methods, two main lineages of T. cruzi have been classified, T. cruzi I and T. cruzi II. Within T. cruzi II, five subdivisions are recognized, T. cruzi IIa-IIe, according to the variability of the ribosomal subunits 24Salpha rRNA and 18S rRNA. In T. rangeli, differences in the organization of the kinetoplast DNA separate two forms denoted T. rangeli KP1+ and KP1-, although differences in the intergenic mini-exon gene and of the small subunit rRNA (SSU rRNA) suggest four subpopulations denoted T. rangeli A, B, C and D. The interactions of these subpopulations of the trypanosomes with different species and populations of Triatominae determine the epidemiology of the human-infecting trypanosomes in Latin America. Often, specific subpopulations of the trypanosomes are transmitted by specific vectors in a particular geographic area. Studies centered on trypanosome-triatomine interaction may allow identification of co-evolutionary processes, which, in turn, could consolidate hypotheses of the evolution and the distribution of T. cruzi/T. rangeli-vectors in America, and they may help to identify the mechanisms that either facilitate or impede the transmission of the parasites in different vector species. Such mechanisms seem to involve intestinal bacteria, especially the symbionts which are needed by the triatomines to complete nymphal development and to produce eggs. Development of the symbionts is regulated by the vector. T. cruzi and T. rangeli interfere with this system and induce the production of antibacterial substances. Whereas T. cruzi is only subpathogenic for the insect host, T. rangeli strongly affects species of the genus Rhodnius and this pathogenicity seems based on a reduction of the number of symbionts.

Role of cellular FKBP52 protein in intracellular trafficking of recombinant adeno-associated virus 2 vectors

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zhao Weihong; Wu Jianqing; Zhong Li

2006-09-30

We have reported that tyrosine-phosphorylated forms of a cellular protein, FKBP52, inhibit the second-strand DNA synthesis of adeno-associated virus 2 (AAV), leading to inefficient transgene expression from recombinant AAV vectors. To further explore the role of FKBP52 in AAV-mediated transduction, we established murine embryo fibroblasts (MEFs) cultures from FKBP52 wild-type (WT), heterozygous (HE), and knockout (KO) mice. Conventional AAV vectors failed to transduce WT MEFs efficiently, and the transduction efficiency was not significantly increased in HE or KO MEFs. AAV vectors failed to traffic efficiently to the nucleus in these cells. Treatment with hydroxyurea (HU) increased the transduction efficiency ofmore » conventional AAV vectors by {approx}25-fold in WT MEFs, but only by {approx}4-fold in KO MEFs. The use of self-complementary AAV (scAAV) vectors, which bypass the requirement of viral second-strand DNA synthesis, revealed that HU treatment increased the transduction efficiency {approx}23-fold in WT MEFs, but only {approx}4-fold in KO MEFs, indicating that the lack of HU treatment-mediated increase in KO MEFs was not due to failure of AAV to undergo viral second-strand DNA synthesis. Following HU treatment, {approx}59% of AAV genomes were present in the nuclear fraction from WT MEFs, but only {approx}28% in KO MEFs, indicating that the pathway by which HU treatment mediates nuclear transport of AAV was impaired in KO MEFs. When KO MEFs were stably transfected with an FKBP52 expression plasmid, HU treatment-mediated increase in the transduction efficiency was restored in these cells, which correlated directly with improved intracellular trafficking. Intact AAV particles were also shown to interact with FKBP52 as well as with dynein, a known cellular protein involved in AAV trafficking. These studies suggest that FKBP52, being a cellular chaperone protein, facilitates intracellular trafficking of AAV, which has implications in the optimal use of recombinant AAV vectors in human gene therapy.« less

Cascading effect of economic globalization on human risks of scrub typhus and tick-borne rickettsial diseases.

PubMed

Kuo, Chi-Chien; Huang, Jing-Lun; Shu, Pei-Yun; Lee, Pei-Lung; Kelt, Douglas A; Wang, Hsi-Chieh

2012-09-01

The increase in global travel and trade has facilitated the dissemination of disease vectors. Globalization can also indirectly affect vector-borne diseases through the liberalization of cross-border trade, which has far-reaching, worldwide effects on agricultural practices and may in turn influence vectors through the modification of the ecological landscape. While the cascading effect of economic globalization on vector-borne diseases, sometimes acting synergistically with regional agricultural policy, could be substantial and have significant economic, agricultural, and public health implications, research into this remains very limited. We evaluated how abandonment of rice paddies in Taiwan after joining the World Trade Organization, along with periodic plowing, an agricultural policy to reduce farm pests in abandoned fields can unexpectedly influence risks to diseases transmitted by ticks and chiggers (larval trombiculid mites), which we collected from their small-mammal hosts. Sampling was limited to abandoned (fallow) and plowed fields due to the challenge of trapping small mammals in flooded rice paddies. Striped field mice (Apodemus agrarius) are the main hosts for both vectors. They harbored six times more ticks and three times more chiggers in fallow than in plowed plots. The proportion of ticks infected with Rickettsia spp. (etiologic agent of spotted fever) was three times higher in fallow plots, while that of Orientia tsutsugamushi (scrub typhus) in chiggers was similar in both treatments. Fallow plots had more ground cover and higher vegetation than plowed ones. Moreover, ticks and chiggers in both field types were dominated by species known to infest humans. Because ticks and chiggers should exhibit very low survival in flooded rice paddies, we propose that farm abandonment in Taiwan, driven by globalization, may have inadvertently led to increased risks of spotted fever and scrub typhus. However, periodic plowing can unintentionally mitigate vector burdens. Economic globalization can have unexpected consequences on disease risk through modification of the agricultural landscape, but the outcome may also be influenced by agricultural policies, calling for further research on vector-borne diseases and their control from broader perspectives.

Impact of the Central Polypurine Tract on the Kinetics of Human Immunodeficiency Virus Type 1 Vector Transduction

PubMed Central

Van Maele, Bénédicte; De Rijck, Jan; De Clercq, Erik; Debyser, Zeger

2003-01-01

Lentiviral vectors derived from human immunodeficiency virus type 1 (HIV-1) show great promise as gene carriers for future gene therapy. Insertion of a fragment containing the central polypurine tract (cPPT) in HIV-1 vector constructs is known to enhance transduction efficiency drastically, reportedly by facilitating the nuclear import of HIV-1 cDNA through a central DNA flap. We have studied the impact of the cPPT on the kinetics of HIV-1 vector transduction by real-time PCR. The kinetics of total HIV-1 DNA, two-long-terminal-repeat (2-LTR) circles, and, by an Alu-PCR, integrated proviral DNA were monitored. About 6 to 12 h after transduction, the total HIV-1 DNA reached a maximum level, followed by a steep decrease. The 2-LTR circles peaked after 24 to 48 h and were diluted upon cell division. Integration of HIV-1 DNA was first detected at 12 h postinfection. When HIV-1 vectors that contained the cPPT were used, DNA synthesis was similar but a threefold higher amount of 2-LTR circles was detected, confirming the impact on nuclear import. Moreover, a 10-fold increase in the amount of integrated DNA was observed in the presence of the cPPT. Only in the absence of the cPPT was a saturation in 2-LTR circle formation seen at a high multiplicity of infection, suggesting a role for the cPPT in overcoming a barrier to the nuclear import of HIV-1 DNA. A major effect of the central DNA flap on the juxtaposition of both LTRs is unlikely, since transduction with HIV-1 vectors containing ectopic cPPT fragments resulted in increased amounts of 2-LTR circles as well as integrated DNA. Inhibitors of transduction by cPPT-containing HIV vectors were also studied by real-time PCR. The reverse transcriptase inhibitor azidothymidine (AZT) and the nonnucleoside reverse transcriptase inhibitor α-APA clearly inhibited viral DNA synthesis, whereas integrase inhibitors such as the diketo acid L-708,906 and the pyranodipyrimidine V-165 specifically inhibited integration. PMID:12663775

Effects of the deletion of early region 4 (E4) open reading frame 1 (orf1), orf1-2, orf1-3 and orf1-4 on virus-host cell interaction, transgene expression, and immunogenicity of replicating adenovirus HIV vaccine vectors.

PubMed

Thomas, Michael A; Song, Rui; Demberg, Thorsten; Vargas-Inchaustegui, Diego A; Venzon, David; Robert-Guroff, Marjorie

2013-01-01

The global health burden engendered by human immunodeficiency virus (HIV)-induced acquired immunodeficiency syndrome (AIDS) is a sobering reminder of the pressing need for a preventative vaccine. In non-human primate models replicating adenovirus (Ad)-HIV/SIV recombinant vaccine vectors have been shown to stimulate potent immune responses culminating in protection against challenge exposures. Nonetheless, an increase in the transgene carrying capacity of these Ad vectors, currently limited to approximately 3000 base pairs, would greatly enhance their utility. Using a replicating, E3-deleted Ad type 5 host range mutant (Ad5 hr) encoding full-length single-chain HIVBaLgp120 linked to the D1 and D2 domains of rhesus macaque CD4 (rhFLSC) we systematically deleted the genes encoding early region 4 open reading frame 1 (E4orf1) through E4orf4. All the Ad-rhFLSC vectors produced similar levels of viral progeny. Cell cycle analysis of infected human and monkey cells revealed no differences in virus-host interaction. The parental and E4-deleted viruses expressed comparable levels of the transgene with kinetics similar to Ad late proteins. Similar levels of cellular immune responses and transgene-specific antibodies were elicited in vaccinated mice. However, differences in recognition of Ad proteins and induced antibody subtypes were observed, suggesting that the E4 gene products might modulate antibody responses by as yet unknown mechanisms. In short, we have improved the transgene carrying capacity by one thousand base pairs while preserving the replicability, levels of transgene expression, and immunogenicity critical to these vaccine vectors. This additional space allows for flexibility in vaccine design that could not be obtained with the current vector and as such should facilitate the goal of improving vaccine efficacy. To the best of our knowledge, this is the first report describing the effects of these E4 deletions on transgene expression and immunogenicity in a replicating Ad vector.

Effects of the Deletion of Early Region 4 (E4) Open Reading Frame 1 (orf1), orf1-2, orf1-3 and orf1-4 on Virus-Host Cell Interaction, Transgene Expression, and Immunogenicity of Replicating Adenovirus HIV Vaccine Vectors

PubMed Central

Thomas, Michael A.; Song, Rui; Demberg, Thorsten; Vargas-Inchaustegui, Diego A.; Venzon, David; Robert-Guroff, Marjorie

2013-01-01

The global health burden engendered by human immunodeficiency virus (HIV)-induced acquired immunodeficiency syndrome (AIDS) is a sobering reminder of the pressing need for a preventative vaccine. In non-human primate models replicating adenovirus (Ad)-HIV/SIV recombinant vaccine vectors have been shown to stimulate potent immune responses culminating in protection against challenge exposures. Nonetheless, an increase in the transgene carrying capacity of these Ad vectors, currently limited to approximately 3000 base pairs, would greatly enhance their utility. Using a replicating, E3-deleted Ad type 5 host range mutant (Ad5 hr) encoding full-length single-chain HIVBaLgp120 linked to the D1 and D2 domains of rhesus macaque CD4 (rhFLSC) we systematically deleted the genes encoding early region 4 open reading frame 1 (E4orf1) through E4orf4. All the Ad-rhFLSC vectors produced similar levels of viral progeny. Cell cycle analysis of infected human and monkey cells revealed no differences in virus-host interaction. The parental and E4-deleted viruses expressed comparable levels of the transgene with kinetics similar to Ad late proteins. Similar levels of cellular immune responses and transgene-specific antibodies were elicited in vaccinated mice. However, differences in recognition of Ad proteins and induced antibody subtypes were observed, suggesting that the E4 gene products might modulate antibody responses by as yet unknown mechanisms. In short, we have improved the transgene carrying capacity by one thousand base pairs while preserving the replicability, levels of transgene expression, and immunogenicity critical to these vaccine vectors. This additional space allows for flexibility in vaccine design that could not be obtained with the current vector and as such should facilitate the goal of improving vaccine efficacy. To the best of our knowledge, this is the first report describing the effects of these E4 deletions on transgene expression and immunogenicity in a replicating Ad vector. PMID:24143187

Microgels produced using microfluidic on-chip polymer blending for controlled released of VEGF encoding lentivectors.

PubMed

Madrigal, Justin L; Sharma, Shonit N; Campbell, Kevin T; Stilhano, Roberta S; Gijsbers, Rik; Silva, Eduardo A

2018-03-15

Alginate hydrogels are widely used as delivery vehicles due to their ability to encapsulate and release a wide range of cargos in a gentle and biocompatible manner. The release of encapsulated therapeutic cargos can be promoted or stunted by adjusting the hydrogel physiochemical properties. However, the release from such systems is often skewed towards burst-release or lengthy retention. To address this, we hypothesized that the overall magnitude of burst release could be adjusted by combining microgels with distinct properties and release behavior. Microgel suspensions were generated using a process we have termed on-chip polymer blending to yield composite suspensions of a range of microgel formulations. In this manner, we studied how alginate percentage and degradation relate to the release of lentivectors. Whereas changes in alginate percentage had a minimal impact on lentivector release, microgel degradation led to a 3-fold increase, and near complete release, over 10 days. Furthermore, by controlling the amount of degradable alginate present within microgels the relative rate of release can be adjusted. A degradable formulation of microgels was used to deliver vascular endothelial growth factor (VEGF)-encoding lentivectors in the chick chorioallantoic membrane (CAM) assay and yielded a proangiogenic response in comparison to the same lentivectors delivered in suspension. The utility of blended microgel suspensions may provide an especially appealing platform for the delivery of lentivectors or similarly sized therapeutics. Genetic therapeutics hold considerable potential for the treatment of diseases and disorders including ischemic cardiovascular diseases. To realize this potential, genetic vectors must be precisely and efficiently delivered to targeted regions of the body. However, conventional methods of delivery do not provide sufficient spatial and temporal control. Here, we demonstrate how alginate microgels provide a basis for developing systems for controlled genetic vector release. We adjust the physiochemical properties of alginate for quicker or slower release, and we demonstrate how combining distinct formulations of microgels can tune the release of the overall composite microgel suspension. These composite suspensions are generated using a straightforward and powerful application of droplet microfluidics which allows for the real-time generation of a composite suspension. Copyright © 2018 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Identification of Conserved Moieties in Metabolic Networks by Graph Theoretical Analysis of Atom Transition Networks

DOE Office of Scientific and Technical Information (OSTI.GOV)

Haraldsdóttir, Hulda S.; Fleming, Ronan M. T.

Conserved moieties are groups of atoms that remain intact in all reactions of a metabolic network. Identification of conserved moieties gives insight into the structure and function of metabolic networks and facilitates metabolic modelling. All moiety conservation relations can be represented as nonnegative integer vectors in the left null space of the stoichiometric matrix corresponding to a biochemical network. Algorithms exist to compute such vectors based only on reaction stoichiometry but their computational complexity has limited their application to relatively small metabolic networks. Moreover, the vectors returned by existing algorithms do not, in general, represent conservation of a specific moietymore » with a defined atomic structure. Here, we show that identification of conserved moieties requires data on reaction atom mappings in addition to stoichiometry. We present a novel method to identify conserved moieties in metabolic networks by graph theoretical analysis of their underlying atom transition networks. Our method returns the exact group of atoms belonging to each conserved moiety as well as the corresponding vector in the left null space of the stoichiometric matrix. It can be implemented as a pipeline of polynomial time algorithms. Our implementation completes in under five minutes on a metabolic network with more than 4,000 mass balanced reactions. The scalability of the method enables extension of existing applications for moiety conservation relations to genome-scale metabolic networks. Finally, we also give examples of new applications made possible by elucidating the atomic structure of conserved moieties.« less

Simple algorithm for improved security in the FDDI protocol

NASA Astrophysics Data System (ADS)

Lundy, G. M.; Jones, Benjamin

1993-02-01

We propose a modification to the Fiber Distributed Data Interface (FDDI) protocol based on a simple algorithm which will improve confidential communication capability. This proposed modification provides a simple and reliable system which exploits some of the inherent security properties in a fiber optic ring network. This method differs from conventional methods in that end to end encryption can be facilitated at the media access control sublayer of the data link layer in the OSI network model. Our method is based on a variation of the bit stream cipher method. The transmitting station takes the intended confidential message and uses a simple modulo two addition operation against an initialization vector. The encrypted message is virtually unbreakable without the initialization vector. None of the stations on the ring will have access to both the encrypted message and the initialization vector except the transmitting and receiving stations. The generation of the initialization vector is unique for each confidential transmission and thus provides a unique approach to the key distribution problem. The FDDI protocol is of particular interest to the military in terms of LAN/MAN implementations. Both the Army and the Navy are considering the standard as the basis for future network systems. A simple and reliable security mechanism with the potential to support realtime communications is a necessary consideration in the implementation of these systems. The proposed method offers several advantages over traditional methods in terms of speed, reliability, and standardization.

Construction of a Shuttle Vector for Heterologous Expression of a Novel Fungal α-Amylase Gene in Aspergillus oryzae.

PubMed

Yin, Yanchen; Mao, Youzhi; Yin, Xiaolie; Gao, Bei; Wei, Dongzhi

2015-07-01

The filamentous fungus Aspergillus oryzae is a well-known expression host used to express homologous and heterologous proteins in a number of industrial applications. To facilitate higher yields of proteins of interest, we constructed the pAsOP vector to express heterologous proteins in A. oryzae. pAsOP carries a selectable marker, pyrG, derived from Aspergillus nidulans, and a strong promoter and a terminator of the amyB gene derived from A. oryzae. pAsOP transformed A. oryzae efficiently via the PEG-CaCl2-mediated transformation method. As proof of concept, green fluorescent protein (GFP) was successfully expressed in A. oryzae transformed by pAsOP-GFP. Additionally, we identified a novel fungal α-amylase (PcAmy) gene from Penicillium sp. and cloned the gene into the vector. After transformation by pAsOPPcAmy, the α-amylase PcAmy from Penicillium sp. was successfully expressed in a heterologous host system for the first time. The α-amylase activity in the A. oryzae transformant was increased by 62.3% compared with the untransformed A. oryzae control. The PcAmy protein produced in the system had an optimum pH of 5.0 and optimum temperature of 30°C. As a cold-adapted enzyme, PcAmy shows potential value in industrial applications because of its high catalytic activity at low temperature. Furthermore, the expression vector reported in this study provides promising utility for further scientific research and biotechnological applications.

Construction of small plasmid vectors for use in genetic improvement of the extremely acidophilic Acidithiobacillus caldus.

PubMed

Meng, Jianzhou; Wang, Huiyan; Liu, Xiangmei; Lin, Jianqun; Pang, Xin; Lin, Jianqiang

2013-10-01

The genetic improvement of biomining bacteria including Acidithiobacillus caldus could facilitate the bioleaching process of sulfur-containing minerals. However, the available vectors for use in A. caldus are very scanty and limited to relatively large broad-host-range IncQ plasmids. In this study, a set of small, mobilizable plasmid vectors (pBBR1MCS-6, pMSD1 and pMSD2) were constructed based on plasmid pBBR1MCS-2, which does not belong to the IncQ, IncW, or IncP groups. The function of the tac promoter on 5.8-kb pMSD2 was determined by inserting a kanamycin-resistant reporter gene. The resulting recombinant pMSD2-Km was successfully transferred by conjugation into A. caldus MTH-04 with transfer frequency of 1.38±0.64×10(-5). The stability and plasmid copy number of pMSD2-Km in A. caldus MTH-04 were 75±2.7% and 5-6 copies per cell, respectively. By inserting an arsABC operon into pMSD2, an arsenic-resistant recombinant pMSD2-As was constructed and transferred into A. caldus MTH-04 by conjugation. The arsenic tolerance of A. caldus MTH-04 containing pMSD2-As was obviously increased up to 45mM of NaAsO2. These vectors could be applied in genetic improvement of A. caldus as well as other bioleaching bacteria. Copyright © 2013 Elsevier GmbH. All rights reserved.

Identification of Conserved Moieties in Metabolic Networks by Graph Theoretical Analysis of Atom Transition Networks

PubMed Central

Haraldsdóttir, Hulda S.; Fleming, Ronan M. T.

2016-01-01

Conserved moieties are groups of atoms that remain intact in all reactions of a metabolic network. Identification of conserved moieties gives insight into the structure and function of metabolic networks and facilitates metabolic modelling. All moiety conservation relations can be represented as nonnegative integer vectors in the left null space of the stoichiometric matrix corresponding to a biochemical network. Algorithms exist to compute such vectors based only on reaction stoichiometry but their computational complexity has limited their application to relatively small metabolic networks. Moreover, the vectors returned by existing algorithms do not, in general, represent conservation of a specific moiety with a defined atomic structure. Here, we show that identification of conserved moieties requires data on reaction atom mappings in addition to stoichiometry. We present a novel method to identify conserved moieties in metabolic networks by graph theoretical analysis of their underlying atom transition networks. Our method returns the exact group of atoms belonging to each conserved moiety as well as the corresponding vector in the left null space of the stoichiometric matrix. It can be implemented as a pipeline of polynomial time algorithms. Our implementation completes in under five minutes on a metabolic network with more than 4,000 mass balanced reactions. The scalability of the method enables extension of existing applications for moiety conservation relations to genome-scale metabolic networks. We also give examples of new applications made possible by elucidating the atomic structure of conserved moieties. PMID:27870845

Cloning-independent plasmid construction for genetic studies in streptococci

PubMed Central

Xie, Zhoujie; Qi, Fengxia; Merritt, Justin

2013-01-01

Shuttle plasmids are among the few routinely utilized tools in the Streptococcus mutans genetic system that still require the use of classical cloning methodologies and intermediate hosts for genetic manipulation. Accordingly, it typically requires considerably less time and effort to introduce mutations onto the S. mutans chromosome than it does to construct shuttle vectors for expressing genes in trans. Occasionally, shuttle vector constructs also exhibit toxicity in E. coli, which prevents their proper assembly. To circumvent these limitations, we modified a prolonged overlap extension PCR (POE-PCR) protocol to facilitate direct plasmid assembly in S. mutans. Using solely PCR, we created the reporter vector pZX7, which contains a single minimal streptococcal replication origin and harbors a spectinomycin resistance cassette and the gusA gene encoding β-glucuronidase. We compared the efficiency of pZX7 assembly using multiple strains of S. mutans and were able to obtain from 5×103 – 2×105 CFU/μg PCR product. Likewise, we used pZX7 to further demonstrate that Streptococcus sanguinis and Streptococcus gordonii are also excellent hosts for cloning-independent plasmid assembly, which suggests that this system is likely to function in numerous other streptococci. Consequently, it should be possible to completely forgo the use of E. coli – Streptococcus shuttle vectors in many streptococcal species, thereby decreasing the time and effort required to assemble constructs and eliminating any toxicity issues associated with intermediate hosts. PMID:23673081

Cloning-independent plasmid construction for genetic studies in streptococci.

PubMed

Xie, Zhoujie; Qi, Fengxia; Merritt, Justin

2013-08-01

Shuttle plasmids are among the few routinely utilized tools in the Streptococcus mutans genetic system that still require the use of classical cloning methodologies and intermediate hosts for genetic manipulation. Accordingly, it typically requires considerably less time and effort to introduce mutations onto the S. mutans chromosome than it does to construct shuttle vectors for expressing genes in trans. Occasionally, shuttle vector constructs also exhibit toxicity in Escherichia coli, which prevents their proper assembly. To circumvent these limitations, we modified a prolonged overlap extension PCR (POE-PCR) protocol to facilitate direct plasmid assembly in S. mutans. Using solely PCR, we created the reporter vector pZX7, which contains a single minimal streptococcal replication origin and harbors a spectinomycin resistance cassette and the gusA gene encoding β-glucuronidase. We compared the efficiency of pZX7 assembly using multiple strains of S. mutans and were able to obtain from 5 × 10³ to 2 × 10⁵ CFU/μg PCR product. Likewise, we used pZX7 to further demonstrate that Streptococcus sanguinis and Streptococcus gordonii are also excellent hosts for cloning-independent plasmid assembly, which suggests that this system is likely to function in numerous other streptococci. Consequently, it should be possible to completely forgo the use of E. coli-Streptococcus shuttle vectors in many streptococcal species, thereby decreasing the time and effort required to assemble constructs and eliminating any toxicity issues associated with intermediate hosts. Copyright © 2013 Elsevier B.V. All rights reserved.

Polymerase chain reaction (PCR) identification of rodent blood meals confirms host sharing by flea vectors of plague.

PubMed

Franklin, Heather A; Stapp, Paul; Cohen, Amybeth

2010-12-01

Elucidating feeding relationships between hosts and parasites remains a significant challenge in studies of the ecology of infectious diseases, especially those involving small or cryptic vectors. Black-tailed prairie dogs (Cynomys ludovicianus) are a species of conservation importance in the North American Great Plains whose populations are extirpated by plague, a flea-vectored, bacterial disease. Using polymerase chain reaction (PCR) assays, we determined that fleas (Oropsylla hirsuta) associated with prairie dogs feed upon northern grasshopper mice (Onychomys leucogaster), a rodent that has been implicated in the transmission and maintenance of plague in prairie-dog colonies. Our results definitively show that grasshopper mice not only share fleas with prairie dogs during plague epizootics, but also provide them with blood meals, offering a mechanism by which the pathogen, Yersinia pestis, may be transmitted between host species and maintained between epizootics. The lack of identifiable host DNA in a significant fraction of engorged Oropsylla hirsuta collected from animals (47%) and prairie-dog burrows (100%) suggests a rapid rate of digestion and feeding that may facilitate disease transmission during epizootics but also complicate efforts to detect feeding on alternative hosts. Combined with other analytical approaches, e.g., stable isotope analysis, molecular genetic techniques can provide novel insights into host-parasite feeding relationships and improve our understanding of the role of alternative hosts in the transmission and maintenance of disease. © 2010 The Society for Vector Ecology.

Identification of Conserved Moieties in Metabolic Networks by Graph Theoretical Analysis of Atom Transition Networks

DOE PAGES

Haraldsdóttir, Hulda S.; Fleming, Ronan M. T.

2016-11-21

Conserved moieties are groups of atoms that remain intact in all reactions of a metabolic network. Identification of conserved moieties gives insight into the structure and function of metabolic networks and facilitates metabolic modelling. All moiety conservation relations can be represented as nonnegative integer vectors in the left null space of the stoichiometric matrix corresponding to a biochemical network. Algorithms exist to compute such vectors based only on reaction stoichiometry but their computational complexity has limited their application to relatively small metabolic networks. Moreover, the vectors returned by existing algorithms do not, in general, represent conservation of a specific moietymore » with a defined atomic structure. Here, we show that identification of conserved moieties requires data on reaction atom mappings in addition to stoichiometry. We present a novel method to identify conserved moieties in metabolic networks by graph theoretical analysis of their underlying atom transition networks. Our method returns the exact group of atoms belonging to each conserved moiety as well as the corresponding vector in the left null space of the stoichiometric matrix. It can be implemented as a pipeline of polynomial time algorithms. Our implementation completes in under five minutes on a metabolic network with more than 4,000 mass balanced reactions. The scalability of the method enables extension of existing applications for moiety conservation relations to genome-scale metabolic networks. Finally, we also give examples of new applications made possible by elucidating the atomic structure of conserved moieties.« less

Identification of Conserved Moieties in Metabolic Networks by Graph Theoretical Analysis of Atom Transition Networks.

PubMed

Haraldsdóttir, Hulda S; Fleming, Ronan M T

2016-11-01

Conserved moieties are groups of atoms that remain intact in all reactions of a metabolic network. Identification of conserved moieties gives insight into the structure and function of metabolic networks and facilitates metabolic modelling. All moiety conservation relations can be represented as nonnegative integer vectors in the left null space of the stoichiometric matrix corresponding to a biochemical network. Algorithms exist to compute such vectors based only on reaction stoichiometry but their computational complexity has limited their application to relatively small metabolic networks. Moreover, the vectors returned by existing algorithms do not, in general, represent conservation of a specific moiety with a defined atomic structure. Here, we show that identification of conserved moieties requires data on reaction atom mappings in addition to stoichiometry. We present a novel method to identify conserved moieties in metabolic networks by graph theoretical analysis of their underlying atom transition networks. Our method returns the exact group of atoms belonging to each conserved moiety as well as the corresponding vector in the left null space of the stoichiometric matrix. It can be implemented as a pipeline of polynomial time algorithms. Our implementation completes in under five minutes on a metabolic network with more than 4,000 mass balanced reactions. The scalability of the method enables extension of existing applications for moiety conservation relations to genome-scale metabolic networks. We also give examples of new applications made possible by elucidating the atomic structure of conserved moieties.

Observation of Interactions in Adolescent Group Therapy: A Mixed Methods Study

PubMed Central

Arias-Pujol, Eulàlia; Anguera, M. Teresa

2017-01-01

Group psychotherapy is a useful clinical practice for adolescents with mental health issues. Groups typically consist of young people of similar ages but with different personalities, and this results in a complex communication network. The goal of group psychoanalytic psychotherapy is to improve participants' mentalization abilities, facilitating interactions between peers and their therapist in a safe, containing environment. The main aim of this study was to analyze conversation turn-taking between a lead therapist, a co-therapist, and six adolescents over the course of 24 treatment sessions divided into four blocks over 8 months. We employed a mixed-methods design based on systematic observation, which we consider to be a mixed method itself, as the qualitative data collected in the initial observation phase is transformed into quantitative data and subsequently interpreted qualitatively with the aid of clinical vignettes. The observational methodology design was nomothetic, follow-up, and multidimensional. The choice of methodology is justified as we used an ad-hoc observation instrument combining a field format and a category system. Interobserver agreement was analyzed quantitatively by Cohen's kappa using the free QSEQ5 software program. Once we had confirmed the reliability of the data, these were analyzed by polar coordinate analysis, which is a powerful data reduction technique that provides a vector representation of relationships between categories. The results show significant relationships between the therapist and (1) the activation of turn-taking by the participants and the co-therapist and silence and (2) conversation-facilitating interventions and interventions designed to improve mentalization abilities. Detailed analysis of questions demonstrating interest in others showed how the communication changed from radial interactions stemming from the therapist at the beginning of therapy to circular interactions half way through. Repetition was found to be a powerful conversation facilitator. The results also illustrate the role of the therapist, who (1) did not facilitate interventions by all participants equally, (2) encouraged turn-taking from more inhibited members of the group, (3) stimulated conversation from the early stages of therapy, and (4) favored mentalization toward the end. Despite its complexity, polar coordinate analysis produces easy-to-interpret results in the form of vector maps. PMID:28790937

Enhanced Central Nervous System Transduction with Lentiviral Vectors Pseudotyped with RVG/HIV-1gp41 Chimeric Envelope Glycoproteins

PubMed Central

Trabalza, Antonio; Eleftheriadou, Ioanna; Sgourou, Argyro; Liao, Ting-Yi; Patsali, Petros; Lee, Heyne

2014-01-01

ABSTRACT To investigate the potential benefits which may arise from pseudotyping the HIV-1 lentiviral vector with its homologous gp41 envelope glycoprotein (GP) cytoplasmic tail (CT), we created chimeric RVG/HIV-1gp41 GPs composed of the extracellular and transmembrane sequences of RVG and either the full-length gp41 CT or C terminus gp41 truncations sequentially removing existing conserved motifs. Lentiviruses (LVs) pseudotyped with the chimeric GPs were evaluated in terms of particle release (physical titer), biological titers, infectivity, and in vivo central nervous system (CNS) transduction. We report here that LVs carrying shorter CTs expressed higher levels of envelope GP and showed a higher average infectivity than those bearing full-length GPs. Interestingly, complete removal of GP CT led to vectors with the highest transduction efficiency. Removal of all C-terminal gp41 CT conserved motifs, leaving just 17 amino acids (aa), appeared to preserve infectivity and resulted in a significantly increased physical titer. Furthermore, incorporation of these 17 aa in the RVG CT notably enhanced the physical titer. In vivo stereotaxic delivery of LV vectors exhibiting the best in vitro titers into rodent striatum facilitated efficient transduction of the CNS at the site of injection. A particular observation was the improved retrograde transduction of neurons in connected distal sites that resulted from the chimeric envelope R5 which included the “Kennedy” sequence (Ken) and lentivirus lytic peptide 2 (LLP2) conserved motifs in the CT, and although it did not exhibit a comparable high titer upon pseudotyping, it led to a significant increase in distal retrograde transduction of neurons. IMPORTANCE In this study, we have produced novel chimeric envelopes bearing the extracellular domain of rabies fused to the cytoplasmic tail (CT) of gp41 and pseudotyped lentiviral vectors with them. Here we report novel effects on the transduction efficiency and physical titer of these vectors, depending on CT length and context. We also managed to achieve increased neuronal transduction in vivo in the rodent CNS, thus demonstrating that the efficiency of these vectors can be enhanced following merely CT manipulation. We believe that this paper is a novel contribution to the field and opens the way for further attempts to surface engineer lentiviral vectors and make them more amenable for applications in human disease. PMID:24371049

A Translational Pathway Toward a Clinical Trial Using the Second-Generation AAV Micro-Dystrophin Vector

DTIC Science & Technology

2016-09-01

minidystrophin gene (a gift from Dr Jeffrey Chamberlain at the University of Washington, Seattle, WA) and the bovine growth hormone polyadenylation...full-length micro-dystrophin protein. Dys-2 is a short peptide in the wild-type full-length dystrophin. It can be recognized by the Dys-2...muscle. In one approach, a muscle homing peptide is inserted on the surface of the capsid to facilitate the entry of AAV into muscle cells. In the

Gene therapy: charting a future course--summary of a National Institutes of Health Workshop, April 12, 2013.

PubMed

O'Reilly, Marina; Federoff, Howard J; Fong, Yuman; Kohn, Donald B; Patterson, Amy P; Ahmed, Nabil; Asokan, Aravind; Boye, Shannon E; Crystal, Ronald G; De Oliveira, Satiro; Gargiulo, Linda; Harper, Scott Q; Ikeda, Yasuhiro; Jambou, Robert; Montgomery, Maureen; Prograis, Lawrence; Rosenthal, Eugene; Sterman, Daniel H; Vandenberghe, Luk H; Zoloth, Laurie; Abedi, Mehrdad; Adair, Jennifer; Adusumilli, Prasad S; Goins, William F; Gray, Jhanelle; Monahan, Paul; Popplewell, Leslie; Sena-Esteves, Miguel; Tannous, Bakhos; Weber, Thomas; Wierda, William; Gopal-Srivastava, Rashmi; McDonald, Cheryl L; Rosenblum, Daniel; Corrigan-Curay, Jacqueline

2014-06-01

Recently, the gene therapy field has begun to experience clinical successes in a number of different diseases using various approaches and vectors. The workshop Gene Therapy: Charting a Future Course, sponsored by the National Institutes of Health (NIH) Office of Biotechnology Activities, brought together early and mid-career researchers to discuss the key scientific challenges and opportunities, ethical and communication issues, and NIH and foundation resources available to facilitate further clinical advances.

Gene Therapy: Charting a Future Course—Summary of a National Institutes of Health Workshop, April 12, 2013

PubMed Central

O'Reilly, Marina; Federoff, Howard J.; Fong, Yuman; Kohn, Donald B.; Patterson, Amy P.; Ahmed, Nabil; Asokan, Aravind; Boye, Shannon E.; Crystal, Ronald G.; De Oliveira, Satiro; Gargiulo, Linda; Harper, Scott Q.; Ikeda, Yasuhiro; Jambou, Robert; Montgomery, Maureen; Prograis, Lawrence; Rosenthal, Eugene; Sterman, Daniel H.; Vandenberghe, Luk H.; Zoloth, Laurie; Abedi, Mehrdad; Adair, Jennifer; Adusumilli, Prasad S.; Goins, William F.; Gray, Jhanelle; Monahan, Paul; Popplewell, Leslie; Sena-Esteves, Miguel; Tannous, Bakhos; Weber, Thomas; Wierda, William; Gopal-Srivastava, Rashmi; McDonald, Cheryl L.; Rosenblum, Daniel

2014-01-01

Abstract Recently, the gene therapy field has begun to experience clinical successes in a number of different diseases using various approaches and vectors. The workshop Gene Therapy: Charting a Future Course, sponsored by the National Institutes of Health (NIH) Office of Biotechnology Activities, brought together early and mid-career researchers to discuss the key scientific challenges and opportunities, ethical and communication issues, and NIH and foundation resources available to facilitate further clinical advances. PMID:24773122

Project ISIAH - Experiment on the effects of micro-gravity on hornets' nest building and activity

NASA Astrophysics Data System (ADS)

Brull, Lily

1992-10-01

An Israel Space Agency Investigation About Hornets (ISIAH) aimed at determining whether hornets are capable of retaining their unique ability of orientation under microgravity conditions is described. The Oriental Hornets used in the experiment are capable of building combs in the direction of the gravitational vector and detecting minute changes in gravitational force. Data obtained may be used to facilitate human adaptation to space conditions as well as rehabilitation after returning to earth.

NATO’s Warsaw Summit: In Brief

DTIC Science & Technology

2016-11-14

Congressional Research Service 9 met the NATO guideline to spend 20% of defense budgets on new equipment and defense modernization and 23 allies spent... budget for FY2017, the Administration requested a fourfold increase in funding for the ERI, primarily to enable “a quicker and more robust response...European allies’ commitment given continued strains on European defense budgets . In addition, some analysts express concern about an apparently growing

Research for the Community: Bates Students Learn as They Work Magic for Lewiston and Auburn

ERIC Educational Resources Information Center

Harward, Donald W.

2001-01-01

When a fire breaks out in the twin cities of Lewiston and Auburn, Maine, the firefighter response time is just a bit quicker these days, thanks to student research at Bates College. When the alarm sounds, firefighters click on a computer map that tells them the size of the burning structure, the quickest route to the fire, the closest hydrant and…

Afghanistan 2012 and Beyond: Clearing, Holding, and Building with Transition Teams

DTIC Science & Technology

2012-05-10

Afghan population, towards achieving those goals. However, given the time already spent in Afghanistan, the constant changes of command and rotation...messages from the White House and apathy of the Afghan government, the pressure is on commanders to expedite the COIN strategy. One problem that...prevents commanders from moving quicker towards transitioning to ANSF is that the definition of success in Afghanistan is difficult to define. In 2009

Lean Manufacturing Improves Emergency Department Throughput and Patient Satisfaction.

PubMed

Kane, Marlena; Chui, Kristen; Rimicci, Janet; Callagy, Patrice; Hereford, James; Shen, Sam; Norris, Robert; Pickham, David

2015-09-01

A multidisciplinary team led by nursing leadership and physicians developed a plan to meet increasing demand and improve the patient experience in the ED without expanding the department's current resources. The approach included Lean tools and engaged frontline staff and physicians. Applying Lean management principles resulted in quicker service, improved patient satisfaction, increased capacity, and reduced resource utilization. Incorporating continuous daily management is necessary for sustainment of continuous improvement activities.

Comparative study of the ''Misgav Ladach'' and traditional Pfannenstiel surgical techniques for cesarean section.

PubMed

Belci, D; Kos, M; Zoricić, D; Kuharić, L; Slivar, A; Begić-Razem, E; Grdinić, I

2007-06-01

The aim of this study was to evaluate the advantages of the Misgav Ladach surgical technique compared to traditional cesarean section. A prospective randomized trial of 111 women undergoing cesarean section was carried out in the Pula General Hospital. Forty-nine operations were performed using the Pfannenstiel method of cesarean section, 55 by the Misgav Ladach method and 7 by lower midline laparotomy. It was proved that the cases where the Misgav Ladach method was implemented, compared to the Pfannenstiel method, showed a significantly shorter delivery/extraction and operative time (P=0.0009), the incision pain on the second postoperative day was significantly lower (0.021), we recorded a quicker stand up and walking time (P=0.013), significantly fewer analgesic injections and a shorter duration of analgesia were required (P=0.0009) and the bowel function was restored to normal sooner (P=0.001). The Misgav Ladach method of cesarean section has advantages over the Pfannenstiel method in so far as it is significantly quicker to perform, with diminished postoperative pain and less use of postoperative analgesics. The recovery of physiologic function is faster. No differences were found in intraoperative bleeding, maternal morbidity, scar appearance, uterus postoperative involution and the assessment of the inflammation response to the operative technique.

Differential Effects of 7 and 16 Groups of Muscle Relaxation Training Following Repeated Submaximal Intensity Exercise in Young Football Players.

PubMed

Sharifah Maimunah, S M P; Hashim, H A

2016-02-01

This study compares two versions of progressive muscle relaxation (PMR) training (7 and 16 muscle groups) on oxygen consumption (VO2), heart rates, rating of perceived exertion and choice reaction time. Football (soccer) players (N = 26; M age = 13.4 yr., SD = 0.5) were randomly assigned to either 7 muscle groups PMR, 16 muscle groups PMR, or a control group. PMR training requires the participants to tense a muscle, hold the muscle contraction, and then relax it. Measurement was conducted prior to and after the completion of 12 sessions of PMR. The dependent variables were measured following four bouts of intermittent exercise consisting of 12 min. of running at 60% VO2max for 10 min. followed by running at 90% VO2max for 2 min. with a 3-min. rest for each bout. Lower VO2, heart rate, perceived exertion, and quicker reaction time were expected in both relaxation groups compared to the control group. The results revealed a significant reduction in heart rates and choice reaction time for both relaxation groups, but the longer version produced significantly quicker choice reaction time. © The Author(s) 2016.

Study on design of light-weight super-abrasive wheel

NASA Astrophysics Data System (ADS)

Nohara, K.; Yanagihara, K.; Ogawa, M.

2018-01-01

Fixed-abrasive tool, also called a grinding wheel, is produced by furnacing abrasive compound which contains abrasive grains and binding powder such as vitrified materials or resins. Fixed-abrasive tool is installed on spindle of grinding machine. And it is given 1,800-2,000 min-1 of spindle rotation for the usage. The centrifugal fracture of the compound of fixed- abrasive tool is one of the careful respects in designing. In recent years, however, super-abrasive wheel as a fixed-abrasive tool has been developed and applied widely. One of the most characteristic respects is that metal is applied for the body of grinding-wheel. The strength to hold abrasive grain and the rigidity of wheel become stronger than those of general grinding wheel, also the lifespan of fixed-abrasive tool becomes longer. The weight of fixed-abrasive tool, however, becomes heavier. Therefore, when the super-abrasive wheel is used, the power consumption of spindle motor becomes larger. It also becomes difficult for the grinding-wheel to respond to sudden acceleration or deceleration. Thus, in order to reduce power consumption in grinding and to obtain quicker frequency response of super-abrasive wheel, the new wheel design is proposed. The design accomplishes 46% weight reduction. Acceleration that is one second quicker than that of conventional grinding wheel is obtained.

Salivary habituation to food stimuli in successful weight loss maintainers, obese and normal-weight adults

PubMed Central

Bond, DS; Raynor, HA; McCaffery, JM; Wing, RR

2017-01-01

Objective Research shows that slower habituation of salivary responses to food stimuli is related to greater energy intake and that obese (Ob) individuals habituate slower than those of normal weight (NW). No study has examined habituation rates in weight loss maintainers (WLMs) who have reduced from obese to normal weight, relative to those who are Ob or NW. Design Salivation to two baseline water trials and 10 lemon-flavored lollipop trials were studied in 14 WLMs, 15 Ob and 18 NW individuals comparable in age, gender and ethnicity. Linear mixed models were used to compare WLMs with Ob and NW groups. Results Salivation in the WLM and NW groups decreased significantly (for both P <0.005) across trials, indicative of habituation. Salivary responses in the Ob group did not habituate (P=0.46). When compared with Ob group, WLMs showed a quicker reduction in salivation (P<0.05). WLM and NW groups did not differ in habituation rate (P=0.49). Conclusions WLMs have habituation rates that are comparable to NW individuals without previous history of obesity, and show quicker habituation than those who are currently obese. These results suggest that physiological responses to food may ‘normalize’ with successful weight loss maintenance. PMID:20010900

Enhanced appetitive learning and reversal learning in a mouse model for Prader-Willi syndrome.

PubMed

Relkovic, Dinko; Humby, Trevor; Hagan, Jim J; Wilkinson, Lawrence S; Isles, Anthony R

2012-06-01

Prader-Willi syndrome (PWS) is caused by lack of paternally derived gene expression from the imprinted gene cluster on human chromosome 15q11-q13. PWS is characterized by severe hypotonia, a failure to thrive in infancy and, on emerging from infancy, evidence of learning disabilities and overeating behavior due to an abnormal satiety response and increased motivation by food. We have previously shown that an imprinting center deletion mouse model (PWS-IC) is quicker to acquire a preference for, and consume more of a palatable food. Here we examined how the use of this palatable food as a reinforcer influences learning in PWS-IC mice performing a simple appetitive learning task. On a nonspatial maze-based task, PWS-IC mice acquired criteria much quicker, making fewer errors during initial acquisition and also reversal learning. A manipulation where the reinforcer was devalued impaired wild-type performance but had no effect on PWS-IC mice. This suggests that increased motivation for the reinforcer in PWS-IC mice may underlie their enhanced learning. This supports previous findings in PWS patients and is the first behavioral study of an animal model of PWS in which the motivation of behavior by food rewards has been examined. © 2012 American Psychological Association

Elastic and acoustic wavefield decompositions and application to reverse time migrations

NASA Astrophysics Data System (ADS)

Wang, Wenlong

P- and S-waves coexist in elastic wavefields, and separation between them is an essential step in elastic reverse-time migrations (RTMs). Unlike the traditional separation methods that use curl and divergence operators, which do not preserve the wavefield vector component information, we propose and compare two vector decomposition methods, which preserve the same vector components that exist in the input elastic wavefield. The amplitude and phase information is automatically preserved, so no amplitude or phase corrections are required. The decoupled propagation method is extended from elastic to viscoelastic wavefields. To use the decomposed P and S vector wavefields and generate PP and PS images, we create a new 2D migration context for isotropic, elastic RTM which includes PS vector decomposition; the propagation directions of both incident and reflected P- and S-waves are calculated directly from the stress and particle velocity definitions of the decomposed P- and S-wave Poynting vectors. Then an excitation-amplitude image condition that scales the receiver wavelet by the source vector magnitude produces angle-dependent images of PP and PS reflection coefficients with the correct polarities, polarization, and amplitudes. It thus simplifies the process of obtaining PP and PS angle-domain common-image gathers (ADCIGs); it is less effort to generate ADCIGs from vector data than from scalar data. Besides P- and S-waves decomposition, separations of up- and down-going waves are also a part of processing of multi-component recorded data and propagating wavefields. A complex trace based up/down separation approach is extended from acoustic to elastic, and combined with P- and S-wave decomposition by decoupled propagation. This eliminates the need for a Fourier transform over time, thereby significantly reducing the storage cost and improving computational efficiency. Wavefield decomposition is applied to both synthetic elastic VSP data and propagating wavefield snapshots. Poynting vectors obtained from the particle-velocity and stress fields after P/S and up/down decompositions are much more accurate than those without. The up/down separation algorithm is also applicable in acoustic RTMs, where both (forward-time extrapolated) source and (reverse-time extrapolated) receiver wavefields are decomposed into up-going and down-going parts. Together with the crosscorrelation imaging condition, four images (down-up, up-down, up-up and down-down) are generated, which facilitate the analysis of artifacts and the imaging ability of the four images. Artifacts may exist in all the decomposed images, but their positions and types are different. The causes of artifacts in different images are explained and illustrated with sketches and numerical tests.

Modified mesenchymal stem cells using miRNA transduction alter lung injury in a bleomycin model.

PubMed

Huleihel, Luai; Sellares, Jacobo; Cardenes, Nayra; Álvarez, Diana; Faner, Rosa; Sakamoto, Koji; Yu, Guoying; Kapetanaki, Maria G; Kaminski, Naftali; Rojas, Mauricio

2017-07-01

Although different preclinical models have demonstrated a favorable role for bone marrow-derived mesenchymal stem cells (B-MSC) in preventing fibrosis, this protective effect is not observed with late administration of these cells, when fibrotic changes are consolidated. We sought to investigate whether the late administration of B-MSCs overexpressing microRNAs (miRNAs) let-7d (antifibrotic) or miR-154 (profibrotic) could alter lung fibrosis in a murine bleomycin model. Using lentiviral vectors, we transduced miRNAs (let-7d or miR-154) or a control sequence into human B-MSCs. Overexpression of let-7d or miR-154 was associated with changes in the mesenchymal properties of B-MSCs and in their cytokine expression. Modified B-MSCs were intravenously administered to mice at day 7 after bleomycin instillation, and the mice were euthanized at day 14 Bleomycin-injured animals that were treated with let-7d cells were found to recover quicker from the initial weight loss compared with the other treatment groups. Interestingly, animals treated with miR-154 cells had the lowest survival rate. Although a slight reduction in collagen mRNA levels was observed in lung tissue from let-7d mice, no significant differences were observed in Ashcroft score and OH-proline. However, the distinctive expression in cytokines and CD45-positive cells in the lung suggests that the differential effects observed in both miRNA mice groups were related to an effect on the immunomodulation function. Our results establish the use of miRNA-modified mesenchymal stem cells as a potential future research in lung fibrosis. Copyright © 2017 the American Physiological Society.

Differential tumor biological role of the tumor suppressor KAI1 and its splice variant in human breast cancer cells

PubMed Central

Miller, Julia; Dreyer, Tobias F.; Bächer, Anne Sophie; Sinner, Eva-Kathrin; Heinrich, Christine; Benge, Anke; Gross, Eva; Preis, Sarah; Rother, Jan; Roberts, Anthony; Nelles, Gabriele; Miteva, Tzenka; Reuning, Ute

2018-01-01

The tetraspanin and tumor suppressor KAI1 is downregulated or lost in many cancers which correlates with poor prognosis. KAI1 acts via physical/functional crosstalk with other membrane receptors. Also, a splice variant of KAI1 (KAI1-SP) has been identified indicative of poor prognosis. We here characterized differential effects of the two KAI1 variants on tumor biological events involving integrin (αvß3) and/or epidermal growth factor receptor (EGF-R). In MDA-MB-231 and -435 breast cancer cells, differential effects were documented on the expression levels of the tumor biologically relevant integrin αvß3 which colocalized with KAI1-WT but not with KAI1-SP. Cellular motility was assessed by video image processing, including motion detection and vector analysis for the quantification and visualization of cell motion parameters. In MDA-MB-231 cells, KAI1-SP provoked a quicker wound gap closure and higher closure rates than KAI1-WT, also reflected by different velocities and average motion amplitudes of singular cells. KAI1-SP induced highest cell motion adjacent to the wound gap borders, whereas in MDA-MB-435 cells a comparable induction of both KAI1 variants was noticed. Moreover, while KAI1-WT reduced cell growth, KAI1-SP significantly increased it going along with a pronounced EGF-R upregulation. KAI1-SP-induced cell migration and proliferation was accompanied by the activation of the focal adhesion and Src kinase. Our findings suggest that splicing of KAI1 does not only abrogate its tumor suppressive functions, but even more, promotes tumor biological effects in favor of cancer progression and metastasis. PMID:29464079

Who Uses the Internet as a Source of Nutrition and Dietary Information? An Australian Population Perspective

PubMed Central

2015-01-01

Background The Internet contains a plethora of nutrition information. Health organizations are increasingly using the Internet to deliver population-wide health information and interventions. Effective interventions identify their target population and their needs; however, little is known about use of the Internet as a source of nutrition information. Objective The aim was to assess the change in prevalence and demographic characteristics of Western Australian adults accessing the Internet as a source of nutrition information and identify specific information needs. Methods Data were pooled from the Western Australian Department of Health’s 3-yearly Nutrition Monitoring Survey Series telephone survey between 1995 and 2012 of 7044 participants aged 18 to 64 years. Outcome variables were the main sources of nutrition information used in the last year and yes/no responses to 4 suggestions to what would make it easier to eat a healthy diet. Sociodemographic variables were collected. Results The proportion of respondents using the Internet for nutrition information increased from <1% in 1995-2001 to 9.1% in 2004 and 33.7% in 2012. Compared to 2004, logistic regression showed that the odds of using the Internet for this information increased significantly in 2009 (OR 2.84, 95% CI 2.07-3.88) and 2012 (OR 5.20, 95% CI 3.86-7.02, P<.001). Respondents using the Internet as a source were more likely to be female (OR 1.30, 95% CI 1.05-1.60, P=.02), live in a metropolitan area (OR 1.26, 95% CI 1.03-1.54, P=.03), born in countries other than Australia/UK/Ireland (OR 1.41, 95% CI 1.07-1.85, P=.02), more educated (university: OR 2.46, 95% CI 1.77-3.42, P<.001), and were less likely to be older (55-64 years: OR 0.38, 95% CI 0.25-0.57, P<.001). The majority of respondents agreed the following information would assist them to make healthier choices: more ways to prepare healthy foods (72.0%, 95% CI 70.7-73.3), quicker ways to prepare healthy foods (79.0%, 95% CI 77.8-80.1), how to choose healthy foods (68.8%, 95% CI 67.5-70.1), and knowing more about cooking (54.7%, 95% CI 53.3-56.1). Those using the Internet for nutrition information were more likely than nonusers to want to know quicker ways to prepare healthy foods (83.0% vs 78.1%, P=.005) and information on choosing healthy foods (76.3% vs 67.3%, P<.001). Conclusions Use of the Internet as a main source of nutrition information has grown rapidly since 2004; one-third of Western Australian adults reported using the Internet for this purpose in 2012. Information on preparing healthy foods (ideas, quicker ways), choosing ingredients, and knowing more about cooking would make it easier to eat a healthy diet. For Internet users, emphasis should be on quicker ways and choosing ingredients. These finding have implications for policy makers and practitioners and suggest that traditional health promotion tactics should continue to be used to reach the broader population. PMID:26310192

Who Uses the Internet as a Source of Nutrition and Dietary Information? An Australian Population Perspective.

PubMed

Pollard, Christina Mary; Pulker, Claire Elizabeth; Meng, Xingqiong; Kerr, Deborah Anne; Scott, Jane Anne

2015-08-26

The Internet contains a plethora of nutrition information. Health organizations are increasingly using the Internet to deliver population-wide health information and interventions. Effective interventions identify their target population and their needs; however, little is known about use of the Internet as a source of nutrition information. The aim was to assess the change in prevalence and demographic characteristics of Western Australian adults accessing the Internet as a source of nutrition information and identify specific information needs. Data were pooled from the Western Australian Department of Health's 3-yearly Nutrition Monitoring Survey Series telephone survey between 1995 and 2012 of 7044 participants aged 18 to 64 years. Outcome variables were the main sources of nutrition information used in the last year and yes/no responses to 4 suggestions to what would make it easier to eat a healthy diet. Sociodemographic variables were collected. The proportion of respondents using the Internet for nutrition information increased from <1% in 1995-2001 to 9.1% in 2004 and 33.7% in 2012. Compared to 2004, logistic regression showed that the odds of using the Internet for this information increased significantly in 2009 (OR 2.84, 95% CI 2.07-3.88) and 2012 (OR 5.20, 95% CI 3.86-7.02, P<.001). Respondents using the Internet as a source were more likely to be female (OR 1.30, 95% CI 1.05-1.60, P=.02), live in a metropolitan area (OR 1.26, 95% CI 1.03-1.54, P=.03), born in countries other than Australia/UK/Ireland (OR 1.41, 95% CI 1.07-1.85, P=.02), more educated (university: OR 2.46, 95% CI 1.77-3.42, P<.001), and were less likely to be older (55-64 years: OR 0.38, 95% CI 0.25-0.57, P<.001). The majority of respondents agreed the following information would assist them to make healthier choices: more ways to prepare healthy foods (72.0%, 95% CI 70.7-73.3), quicker ways to prepare healthy foods (79.0%, 95% CI 77.8-80.1), how to choose healthy foods (68.8%, 95% CI 67.5-70.1), and knowing more about cooking (54.7%, 95% CI 53.3-56.1). Those using the Internet for nutrition information were more likely than nonusers to want to know quicker ways to prepare healthy foods (83.0% vs 78.1%, P=.005) and information on choosing healthy foods (76.3% vs 67.3%, P<.001). Use of the Internet as a main source of nutrition information has grown rapidly since 2004; one-third of Western Australian adults reported using the Internet for this purpose in 2012. Information on preparing healthy foods (ideas, quicker ways), choosing ingredients, and knowing more about cooking would make it easier to eat a healthy diet. For Internet users, emphasis should be on quicker ways and choosing ingredients. These finding have implications for policy makers and practitioners and suggest that traditional health promotion tactics should continue to be used to reach the broader population.

Use of elevator instruments when luxating and extracting teeth in dentistry: clinical techniques

PubMed Central

2017-01-01

In dentistry, elevator instruments are used to luxate teeth, and this technique imparts forces to tooth particles that sever the periodontal ligament around tooth roots inside the socket and expand alveolar bone around tooth particles. These effects can result in extraction of the tooth particles or facilitate systematic forceps extraction of the tooth particles. This article presents basic oral surgery techniques for applying elevators to luxate teeth. Determination of the optimal luxation technique requires understanding of the functions of the straight elevator and the Cryer elevator, the concept of purchase points, how the design elements of elevator working ends and tips influence the functionality of an elevator, application of forces to tooth particles, sectioning teeth at furcations, and bone removal to facilitate luxation. The effectiveness of tooth particle luxation is influenced by elevator tip shape and size, the magnitude and the vectors of forces applied to the tooth particle by the tip, and sectioning and bone removal within the operating field. Controlled extraction procedures are facilitated by a dental operating microscope or the magnification of binocular surgical loupes telescopes, combined with co-axial illumination. PMID:28770164

Automatic classification of 6-month-old infants at familial risk for language-based learning disorder using a support vector machine.

PubMed

Zare, Marzieh; Rezvani, Zahra; Benasich, April A

2016-07-01

This study assesses the ability of a novel, "automatic classification" approach to facilitate identification of infants at highest familial risk for language-learning disorders (LLD) and to provide converging assessments to enable earlier detection of developmental disorders that disrupt language acquisition. Network connectivity measures derived from 62-channel electroencephalogram (EEG) recording were used to identify selected features within two infant groups who differed on LLD risk: infants with a family history of LLD (FH+) and typically-developing infants without such a history (FH-). A support vector machine was deployed; global efficiency and global and local clustering coefficients were computed. A novel minimum spanning tree (MST) approach was also applied. Cross-validation was employed to assess the resultant classification. Infants were classified with about 80% accuracy into FH+ and FH- groups with 89% specificity and precision of 92%. Clustering patterns differed by risk group and MST network analysis suggests that FH+ infants' EEG complexity patterns were significantly different from FH- infants. The automatic classification techniques used here were shown to be both robust and reliable and should provide valuable information when applied to early identification of risk or clinical groups. The ability to identify infants at highest risk for LLD using "automatic classification" strategies is a novel convergent approach that may facilitate earlier diagnosis and remediation. Copyright © 2016 International Federation of Clinical Neurophysiology. Published by Elsevier Ireland Ltd. All rights reserved.

The Impact of Commodity Councils on the Internal Structures of Purchasing Organizations in the United States Air Force

DTIC Science & Technology

2004-12-01

product or service) (Tyndall 133). Purchasing in bulk allows companies to take advantage of volume discounts, while at the same time streamlining...for industry. They included, financial advantage , strategic global sourcing, quicker response to marketplace changes, a paperless environment for...purchasing, and an increased competitive advantage (Richter). He cautioned the Air Force to avoid the trap of focusing only on efficiency in creating

Mission Analysis for Multiple Rendezvous of Near-Earth Asteroids Using Earth Gravity Assist

DTIC Science & Technology

2010-03-01

devices. Finding solutions with this approach leads to a quicker timeline for possible missions since one does not have to wait for the propulsion...in this research. The discussion focuses on their approach to the problem and the applicability to this research. The headings are the titles of... approach the problem utilizing conventional impulsive thrust propulsion systems and utilize data presented from the JPL website for locating the

Smart Microgrid Energy Management Controls for Improved Energy Efficiency and Renewables Integration at DoD Installations

DTIC Science & Technology

2013-04-01

products for energy generation, including solar, wind , and gas turbines , energy storage, power conversion, grid integration, and software for...potential security advantages over centralized systems • DER promote fuel diversity (e.g., biomass, landfill gas, flare gas, wind , solar) and...therefore reduce overall energy price volatility • Renewable DER such as wind and solar photovoltaics provide emissions-free energy • DER offer a quicker

The Effect of Age on the Susceptibility and Severity of Demyelination

DTIC Science & Technology

2015-10-01

Animal facilities (completed) Task 2d- Breed the offspring and ascertain that the transgenic trait is present – (completed) Task 3e... transgenic mouse the consequence was that these mice became sicker much quicker than a cohort of mice with normal levels of neurofascin. Therefore we...disease. In addition we challenged transgenic mice that had greatly diminished amounts of the molecule which links myelin to the axon (neurofascin) to

Technology Strategy in Irregular Warfare: High-Tech Versus Right-Tech

DTIC Science & Technology

2015-12-01

ny-countys-confiscated-gun-policy/. 88 newest AH-64E. It has upgraded engines and rotor blades that enable the attack helicopter to have a quicker...was the extent of their defensive operations. Aircraft, helicopter gunships, armored vehicles, and artillery were directly used by Soviet forces...it into raw numbers, the Soviet Air Force had approximately 6,894 fixed-wing aircraft, and 3,320 helicopters .126 The Soviet Army had five times

Development and Testing of a Dual Accelerometer Vector Sensor for AUV Acoustic Surveys †

PubMed Central

Mantouka, Agni; Felisberto, Paulo; Santos, Paulo; Zabel, Friedrich; Saleiro, Mário; Jesus, Sérgio M.; Sebastião, Luís

2017-01-01

This paper presents the design, manufacturing and testing of a Dual Accelerometer Vector Sensor (DAVS). The device was built within the activities of the WiMUST project, supported under the Horizon 2020 Framework Programme, which aims to improve the efficiency of the methodologies used to perform geophysical acoustic surveys at sea by the use of Autonomous Underwater Vehicles (AUVs). The DAVS has the potential to contribute to this aim in various ways, for example, owing to its spatial filtering capability, it may reduce the amount of post processing by discriminating the bottom from the surface reflections. Additionally, its compact size allows easier integration with AUVs and hence facilitates the vehicle manoeuvrability compared to the classical towed arrays. The present paper is focused on results related to acoustic wave azimuth estimation as an example of its spatial filtering capabilities. The DAVS device consists of two tri-axial accelerometers and one hydrophone moulded in one unit. Sensitivity and directionality of these three sensors were measured in a tank, whilst the direction estimation capabilities of the accelerometers paired with the hydrophone, forming a vector sensor, were evaluated on a Medusa Class AUV, which was sailing around a deployed sound source. Results of these measurements are presented in this paper. PMID:28594342

Development and Testing of a Dual Accelerometer Vector Sensor for AUV Acoustic Surveys.

PubMed

Mantouka, Agni; Felisberto, Paulo; Santos, Paulo; Zabel, Friedrich; Saleiro, Mário; Jesus, Sérgio M; Sebastião, Luís

2017-06-08

This paper presents the design, manufacturing and testing of a Dual Accelerometer Vector Sensor (DAVS). The device was built within the activities of the WiMUST project, supported under the Horizon 2020 Framework Programme, which aims to improve the efficiency of the methodologies used to perform geophysical acoustic surveys at sea by the use of Autonomous Underwater Vehicles (AUVs). The DAVS has the potential to contribute to this aim in various ways, for example, owing to its spatial filtering capability, it may reduce the amount of post processing by discriminating the bottom from the surface reflections. Additionally, its compact size allows easier integration with AUVs and hence facilitates the vehicle manoeuvrability compared to the classical towed arrays. The present paper is focused on results related to acoustic wave azimuth estimation as an example of its spatial filtering capabilities. The DAVS device consists of two tri-axial accelerometers and one hydrophone moulded in one unit. Sensitivity and directionality of these three sensors were measured in a tank, whilst the direction estimation capabilities of the accelerometers paired with the hydrophone, forming a vector sensor, were evaluated on a Medusa Class AUV, which was sailing around a deployed sound source. Results of these measurements are presented in this paper.

You can hide but you have to run: direct detection with vector mediators

NASA Astrophysics Data System (ADS)

D'Eramo, Francesco; Kavanagh, Bradley J.; Panci, Paolo

2016-08-01

We study direct detection in simplified models of Dark Matter (DM) in which interactions with Standard Model (SM) fermions are mediated by a heavy vector boson. We consider fully general, gauge-invariant couplings between the SM, the mediator and both scalar and fermion DM. We account for the evolution of the couplings between the energy scale of the mediator mass and the nuclear energy scale. This running arises from virtual effects of SM particles and its inclusion is not optional. We compare bounds on the mediator mass from direct detection experiments with and without accounting for the running. In some cases the inclusion of these effects changes the bounds by several orders of magnitude, as a consequence of operator mixing which generates new interactions at low energy. We also highlight the importance of these effects when translating LHC limits on the mediator mass into bounds on the direct detection cross section. For an axial-vector mediator, the running can alter the derived bounds on the spin-dependent DM-nucleon cross section by a factor of two or more. Finally, we provide tools to facilitate the inclusion of these effects in future studies: general approximate expressions for the low energy couplings and a public code runDM to evolve the couplings between arbitrary energy scales.

Sex-biased avian host use by arbovirus vectors.

PubMed

Burkett-Cadena, Nathan D; Bingham, Andrea M; Unnasch, Thomas R

2014-11-01

Prevalence of arthropod-borne parasites often differs drastically between host sexes. This sex-related disparity may be related to physiological (primarily hormonal) differences that facilitate or suppress replication of the pathogen in host tissues. Alternately, differences in pathogen prevalence between host sexes may be owing to differential exposure to infected vectors. Here, we report on the use of PCR-based assays recognizing bird sex chromosomes to investigate sex-related patterns of avian host use from field-collected female mosquitoes from Florida, USA. Mosquitoes took more bloodmeals from male birds (64.0% of 308 sexed samples) than female birds (36.0%), deviating significantly from a hypothetical 1:1 sex ratio. In addition, male-biased host use was consistent across mosquito species (Culex erraticus (64.4%); Culex nigripalpus (61.0%) and Culiseta melanura (64.9%)). Our findings support the hypothesis that sex-biased exposure to vector-borne pathogens contributes to disparities in parasite/pathogen prevalence between the sexes. While few studies have yet to investigate sex-biased host use by mosquitoes, the methods used here could be applied to a variety of mosquito-borne disease systems, including those that affect health of humans, domestic animals and wildlife. Understanding the mechanisms that drive sex-based disparities in host use may lead to novel strategies for interrupting pathogen/parasite transmission.

Broad-host-range vector system for synthetic biology and biotechnology in cyanobacteria

PubMed Central

Taton, Arnaud; Unglaub, Federico; Wright, Nicole E.; Zeng, Wei Yue; Paz-Yepes, Javier; Brahamsha, Bianca; Palenik, Brian; Peterson, Todd C.; Haerizadeh, Farzad; Golden, Susan S.; Golden, James W.

2014-01-01

Inspired by the developments of synthetic biology and the need for improved genetic tools to exploit cyanobacteria for the production of renewable bioproducts, we developed a versatile platform for the construction of broad-host-range vector systems. This platform includes the following features: (i) an efficient assembly strategy in which modules released from 3 to 4 donor plasmids or produced by polymerase chain reaction are assembled by isothermal assembly guided by short GC-rich overlap sequences. (ii) A growing library of molecular devices categorized in three major groups: (a) replication and chromosomal integration; (b) antibiotic resistance; (c) functional modules. These modules can be assembled in different combinations to construct a variety of autonomously replicating plasmids and suicide plasmids for gene knockout and knockin. (iii) A web service, the CYANO-VECTOR assembly portal, which was built to organize the various modules, facilitate the in silico construction of plasmids, and encourage the use of this system. This work also resulted in the construction of an improved broad-host-range replicon derived from RSF1010, which replicates in several phylogenetically distinct strains including a new experimental model strain Synechocystis sp. WHSyn, and the characterization of nine antibiotic cassettes, four reporter genes, four promoters, and a ribozyme-based insulator in several diverse cyanobacterial strains. PMID:25074377

Development of Transcriptional Fusions to Assess Leptospira interrogans Promoter Activity

PubMed Central

Cerqueira, Gustavo M.; Souza, Natalie M.; Araújo, Eduardo R.; Barros, Aline T.; Morais, Zenaide M.; Vasconcellos, Sílvio A.; Nascimento, Ana L. T. O.

2011-01-01

Background Leptospirosis is a zoonotic infectious disease that affects both humans and animals. The existing genetic tools for Leptospira spp. have improved our understanding of the biology of this spirochete as well as the interaction of pathogenic leptospires with the mammalian host. However, new tools are necessary to provide novel and useful information to the field. Methodology and Principal Findings A series of promoter-probe vectors carrying a reporter gene encoding green fluorescent protein (GFP) were constructed for use in L. biflexa. They were tested by constructing transcriptional fusions between the lipL41, Leptospiral Immunoglobulin-like A (ligA) and Sphingomielynase 2 (sph2) promoters from L. interrogans and the reporter gene. ligA and sph2 promoters were the most active, in comparison to the lipL41 promoter and the non-induced controls. The results obtained are in agreement with LigA expression from the L. interrogans Fiocruz L1-130 strain. Conclusions The novel vectors facilitated the in vitro evaluation of L. interrogans promoter activity under defined growth conditions which simulate the mammalian host environment. The fluorescence and rt-PCR data obtained closely reflected transcriptional regulation of the promoters, thus demonstrating the suitability of these vectors for assessing promoter activity in L. biflexa. PMID:21445252

A novel approach for dimension reduction of microarray.

PubMed

Aziz, Rabia; Verma, C K; Srivastava, Namita

2017-12-01

This paper proposes a new hybrid search technique for feature (gene) selection (FS) using Independent component analysis (ICA) and Artificial Bee Colony (ABC) called ICA+ABC, to select informative genes based on a Naïve Bayes (NB) algorithm. An important trait of this technique is the optimization of ICA feature vector using ABC. ICA+ABC is a hybrid search algorithm that combines the benefits of extraction approach, to reduce the size of data and wrapper approach, to optimize the reduced feature vectors. This hybrid search technique is facilitated by evaluating the performance of ICA+ABC on six standard gene expression datasets of classification. Extensive experiments were conducted to compare the performance of ICA+ABC with the results obtained from recently published Minimum Redundancy Maximum Relevance (mRMR) +ABC algorithm for NB classifier. Also to check the performance that how ICA+ABC works as feature selection with NB classifier, compared the combination of ICA with popular filter techniques and with other similar bio inspired algorithm such as Genetic Algorithm (GA) and Particle Swarm Optimization (PSO). The result shows that ICA+ABC has a significant ability to generate small subsets of genes from the ICA feature vector, that significantly improve the classification accuracy of NB classifier compared to other previously suggested methods. Copyright © 2017 Elsevier Ltd. All rights reserved.

Anopheles plumbeus (Diptera: Culicidae) in Germany: updated geographic distribution and public health impact of a nuisance and vector mosquito.

PubMed

Heym, Eva C; Kampen, Helge; Fahle, Marcus; Hohenbrink, Tobias L; Schäfer, Mandy; Scheuch, Dorothee E; Walther, Doreen

2017-01-01

The aim of this study was to map the current spatial distribution of Anopheles plumbeus in Germany, a potential vector of malaria parasites and West Nile virus. Reports of mass occurrence and nuisance connected with artificial breeding site usage by this species were analysed. Distribution data were collected from 2011 to 2014 mainly through trapping and submissions of adult mosquito specimens to a citizen science project. In the framework of the latter, additional information was gathered on recent nuisance incidents caused by An. plumbeus, including a longitudinal analysis of mosquito occurrence and the impact of management measures at a nuisance site in south-western Germany. Based on the most comprehensive set of collection data obtained during the last decades, An. plumbeus is shown to be widely distributed over Germany. The data also indicate a continuing extension of the breeding site repertoire of the species from natural to artificial habitats that facilitate mass development. Increasing incidents of persistent nuisance suggest that this mosquito species is rarely diagnosed correctly and managed adequately. As An. plumbeus is both a serious nuisance pest and a potential vector species, awareness of this species and the public health problems linked to it should be raised among pest managers and public health personnel. © 2016 John Wiley & Sons Ltd.

Gene Delivery into Plant Cells for Recombinant Protein Production

PubMed Central

Chen, Qiang

2015-01-01

Recombinant proteins are primarily produced from cultures of mammalian, insect, and bacteria cells. In recent years, the development of deconstructed virus-based vectors has allowed plants to become a viable platform for recombinant protein production, with advantages in versatility, speed, cost, scalability, and safety over the current production paradigms. In this paper, we review the recent progress in the methodology of agroinfiltration, a solution to overcome the challenge of transgene delivery into plant cells for large-scale manufacturing of recombinant proteins. General gene delivery methodologies in plants are first summarized, followed by extensive discussion on the application and scalability of each agroinfiltration method. New development of a spray-based agroinfiltration and its application on field-grown plants is highlighted. The discussion of agroinfiltration vectors focuses on their applications for producing complex and heteromultimeric proteins and is updated with the development of bridge vectors. Progress on agroinfiltration in Nicotiana and non-Nicotiana plant hosts is subsequently showcased in context of their applications for producing high-value human biologics and low-cost and high-volume industrial enzymes. These new advancements in agroinfiltration greatly enhance the robustness and scalability of transgene delivery in plants, facilitating the adoption of plant transient expression systems for manufacturing recombinant proteins with a broad range of applications. PMID:26075275

Transposon mutagenesis of Xylella fastidiosa by electroporation of Tn5 synaptic complexes.

PubMed

Guilhabert, M R; Hoffman, L M; Mills, D A; Kirkpatrick, B C

2001-06-01

Pierce's disease, a lethal disease of grapevine, is caused by Xylella fastidiosa, a gram-negative, xylem-limited bacterium that is transmitted from plant to plant by xylem-feeding insects. Strains of X. fastidiosa also have been associated with diseases that cause tremendous losses in many other economically important plants, including citrus. Although the complete genome sequence of X. fastidiosa has recently been determined, the inability to transform or produce transposon mutants of X. fastidiosa has been a major impediment to understanding pathogen-, plant-, and insect-vector interactions. We evaluated the ability of four different suicide vectors carrying either Tn5 or Tn10 transposons as well as a preformed Tn5 transposase-transposon synaptic complex (transposome) to transpose X. fastidiosa. The four suicide vectors failed to produce any detectable transposition events. Electroporation of transposomes, however, yielded 6 x 10(3) and 4 x 10(3) Tn5 mutants per microg of DNA in two different grapevine strains of X. fastidiosa. Molecular analysis showed that the transposition insertions were single, independent, stable events. Sequence analysis of the Tn5 insertion sites indicated that the transpositions occur randomly in the X. fastidiosa genome. Transposome-mediated mutagenesis should facilitate the identification of X. fastidiosa genes that mediate plant pathogenicity and insect transmission.

Peripheral infrastructure vectors and an extended set of plant parts for the Modular Cloning system

PubMed Central

Kretschmer, Carola; Gruetzner, Ramona; Löfke, Christian; Dagdas, Yasin; Bürstenbinder, Katharina; Marillonnet, Sylvestre

2018-01-01

Standardized DNA assembly strategies facilitate the generation of multigene constructs from collections of building blocks in plant synthetic biology. A common syntax for hierarchical DNA assembly following the Golden Gate principle employing Type IIs restriction endonucleases was recently developed, and underlies the Modular Cloning and GoldenBraid systems. In these systems, transcriptional units and/or multigene constructs are assembled from libraries of standardized building blocks, also referred to as phytobricks, in several hierarchical levels and by iterative Golden Gate reactions. Here, a toolkit containing further modules for the novel DNA assembly standards was developed. Intended for use with Modular Cloning, most modules are also compatible with GoldenBraid. Firstly, a collection of approximately 80 additional phytobricks is provided, comprising e.g. modules for inducible expression systems, promoters or epitope tags. Furthermore, DNA modules were developed for connecting Modular Cloning and Gateway cloning, either for toggling between systems or for standardized Gateway destination vector assembly. Finally, first instances of a “peripheral infrastructure” around Modular Cloning are presented: While available toolkits are designed for the assembly of plant transformation constructs, vectors were created to also use coding sequence-containing phytobricks directly in yeast two hybrid interaction or bacterial infection assays. The presented material will further enhance versatility of hierarchical DNA assembly strategies. PMID:29847550

Engineering nanoparticle-coated bacteria as oral DNA vaccines for cancer immunotherapy.

PubMed

Hu, Qinglian; Wu, Min; Fang, Chun; Cheng, Changyong; Zhao, Mengmeng; Fang, Weihuan; Chu, Paul K; Ping, Yuan; Tang, Guping

2015-04-08

Live attenuated bacteria are of increasing importance in biotechnology and medicine in the emerging field of cancer immunotherapy. Oral DNA vaccination mediated by live attenuated bacteria often suffers from low infection efficiency due to various biological barriers during the infection process. To this end, we herein report, for the first time, a new strategy to engineer cationic nanoparticle-coated bacterial vectors that can efficiently deliver oral DNA vaccine for efficacious cancer immunotherapy. By coating live attenuated bacteria with synthetic nanoparticles self-assembled from cationic polymers and plasmid DNA, the protective nanoparticle coating layer is able to facilitate bacteria to effectively escape phagosomes, significantly enhance the acid tolerance of bacteria in stomach and intestines, and greatly promote dissemination of bacteria into blood circulation after oral administration. Most importantly, oral delivery of DNA vaccines encoding autologous vascular endothelial growth factor receptor 2 (VEGFR2) by this hybrid vector showed remarkable T cell activation and cytokine production. Successful inhibition of tumor growth was also achieved by efficient oral delivery of VEGFR2 with nanoparticle-coated bacterial vectors due to angiogenesis suppression in the tumor vasculature and tumor necrosis. This proof-of-concept work demonstrates that coating live bacterial cells with synthetic nanoparticles represents a promising strategy to engineer efficient and versatile DNA vaccines for the era of immunotherapy.

A simple method of equine limb force vector analysis and its potential applications

PubMed Central

Robinson, Mark A.; Clayton, Hilary M.

2018-01-01

Background Ground reaction forces (GRF) measured during equine gait analysis are typically evaluated by analyzing discrete values obtained from continuous force-time data for the vertical, longitudinal and transverse GRF components. This paper describes a simple, temporo-spatial method of displaying and analyzing sagittal plane GRF vectors. In addition, the application of statistical parametric mapping (SPM) is introduced to analyse differences between contra-lateral fore and hindlimb force-time curves throughout the stance phase. The overall aim of the study was to demonstrate alternative methods of evaluating functional (a)symmetry within horses. Methods GRF and kinematic data were collected from 10 horses trotting over a series of four force plates (120 Hz). The kinematic data were used to determine clean hoof contacts. The stance phase of each hoof was determined using a 50 N threshold. Vertical and longitudinal GRF for each stance phase were plotted both as force-time curves and as force vector diagrams in which vectors originating at the centre of pressure on the force plate were drawn at intervals of 8.3 ms for the duration of stance. Visual evaluation was facilitated by overlay of the vector diagrams for different limbs. Summary vectors representing the magnitude (VecMag) and direction (VecAng) of the mean force over the entire stance phase were superimposed on the force vector diagram. Typical measurements extracted from the force-time curves (peak forces, impulses) were compared with VecMag and VecAng using partial correlation (controlling for speed). Paired samples t-tests (left v. right diagonal pair comparison and high v. low vertical force diagonal pair comparison) were performed on discrete and vector variables using traditional methods and Hotelling’s T2 tests on normalized stance phase data using SPM. Results Evidence from traditional statistical tests suggested that VecMag is more influenced by the vertical force and impulse, whereas VecAng is more influenced by the longitudinal force and impulse. When used to evaluate mean data from the group of ten sound horses, SPM did not identify differences between the left and right contralateral limb pairs or between limb pairs classified according to directional asymmetry. When evaluating a single horse, three periods were identified during which differences in the forces between the left and right forelimbs exceeded the critical threshold (p < .01). Discussion Traditional statistical analysis of 2D GRF peak values, summary vector variables and visual evaluation of force vector diagrams gave harmonious results and both methods identified the same inter-limb asymmetries. As alpha was more tightly controlled using SPM, significance was only found in the individual horse although T2 plots followed the same trends as discrete analysis for the group. Conclusions The techniques of force vector analysis and SPM hold promise for investigations of sidedness and asymmetry in horses. PMID:29492341

Some ways of plants wastes utilization in bioregenerative life support systems

NASA Astrophysics Data System (ADS)

Kovaleva, N. P.; Tikhomirov, A. A.; Tirranen, L. S.; Ushakova, S. A.; Zolotukhin, I. G.; Anischenko, O. V.

In works on experimental modeling of bioregenerative life support systems BLSS carried out at Institute of Biophysics Russian Academy of Science Siberian Branch SB RAS the possibility of increase of a system closure degree under the condition of inedible plant biomass return into the organic matter turnover was demonstrated At the same time when radish inedible biomass was subjected to biological oxidation in soil-like substrate SLS after its drying then wheat straw was subjected to stepwise processing including mushrooms growing stage Mushrooms cultivation facilitated to lignin destruction and quicker straw decomposition On the other hand mushrooms growing required additional technological procedures leading to complication of a technological chain of straw processing The purpose of this work is to study the possibility of exclusion of mushrooms growing stage under straw pretreatment for its further use as an equivalent of radish edible biomass grown on SLS To solve the problem put by the radish cenosis in a conveyer regime was grown The conveyer included radish four ages with the conveyer step equal to 7 days The experiment consisted of two successive stages On the first stage radish was grown without straw addition into SLS control To return mineral elements into SLS the biomass grown was restored in SLS On the second stage inedible radish biomass and wheat straw were returned into SLS in the quantity equivalent to edible biomass The possibility of the method described was estimated according to plant productivity microbiological

Visual processing of emotional expressions in mixed anxious-depressed subclinical state: an event-related potential study on a female sample.

PubMed

Rossignol, M; Philippot, P; Crommelinck, M; Campanella, S

2008-10-01

Controversy remains about the existence and the nature of a specific bias in emotional facial expression processing in mixed anxious-depressed state (MAD). Event-related potentials were recorded in the following three types of groups defined by the Spielberger state and trait anxiety inventory (STAI) and the Beck depression inventory (BDI): a group of anxious participants (n=12), a group of participants with depressive and anxious tendencies (n=12), and a control group (n=12). Participants were confronted with a visual oddball task in which they had to detect, as quickly as possible, deviant faces amongst a train of standard neutral faces. Deviant stimuli changed either on identity, or on emotion (happy or sad expression). Anxiety facilitated emotional processing and the two anxious groups produced quicker responses than control participants; these effects were correlated with an earlier decisional wave (P3b) for anxious participants. Mixed anxious-depressed participants showed enhanced visual processing of deviant stimuli and produced higher amplitude in attentional complex (N2b/P3a), both for identity and emotional trials. P3a was also particularly increased for emotional faces in this group. Anxious state mainly influenced later decision processes (shorter latency of P3b), whereas mixed anxious-depressed state acted on earlier steps of emotional processing (enhanced N2b/P3a complex). Mixed anxious-depressed individuals seemed more reactive to any visual change, particularly emotional change, without displaying any valence bias.

Fluorescent protein vectors for pancreatic islet cell identification in live-cell imaging.

PubMed

Shuai, Hongyan; Xu, Yunjian; Yu, Qian; Gylfe, Erik; Tengholm, Anders

2016-10-01

The islets of Langerhans contain different types of endocrine cells, which are crucial for glucose homeostasis. β- and α-cells that release insulin and glucagon, respectively, are most abundant, whereas somatostatin-producing δ-cells and particularly pancreatic polypeptide-releasing PP-cells are more scarce. Studies of islet cell function are hampered by difficulties to identify the different cell types, especially in live-cell imaging experiments when immunostaining is unsuitable. The aim of the present study was to create a set of vectors for fluorescent protein expression with cell-type-specific promoters and evaluate their applicability in functional islet imaging. We constructed six adenoviral vectors for expression of red and green fluorescent proteins controlled by the insulin, preproglucagon, somatostatin, or pancreatic polypeptide promoters. After transduction of mouse and human islets or dispersed islet cells, a majority of the fluorescent cells also immunostained for the appropriate hormone. Recordings of the sub-plasma membrane Ca(2+) and cAMP concentrations with a fluorescent indicator and a protein biosensor, respectively, showed that labeled cells respond to glucose and other modulators of secretion and revealed a striking variability in Ca(2+) signaling among α-cells. The measurements allowed comparison of the phase relationship of Ca(2+) oscillations between different types of cells within intact islets. We conclude that the fluorescent protein vectors allow easy identification of specific islet cell types and can be used in live-cell imaging together with organic dyes and genetically encoded biosensors. This approach will facilitate studies of normal islet physiology and help to clarify molecular defects and disturbed cell interactions in diabetic islets.

AAV-mediated targeting of gene expression to the peri-infarct region in rat cortical stroke model.

PubMed

Mätlik, Kert; Abo-Ramadan, Usama; Harvey, Brandon K; Arumäe, Urmas; Airavaara, Mikko

2014-10-30

For stroke patients the recovery of cognitive and behavioral functions is often incomplete. Functional recovery is thought to be mediated largely by connectivity rearrangements in the peri-infarct region. A method for manipulating gene expression in this region would be useful for identifying new recovery-enhancing treatments. We have characterized a way of targeting adeno-associated virus (AAV) vectors to the peri-infarct region of cortical ischemic lesion in rats 2days after middle cerebral artery occlusion (MCAo). We used magnetic resonance imaging (MRI) to show that the altered properties of post-ischemic brain tissue facilitate the spreading of intrastriatally injected nanoparticles toward the infarct. We show that subcortical injection of green fluorescent protein-encoding dsAAV7-GFP resulted in transduction of cells in and around the white matter tract underlying the lesion, and in the cortex proximal to the lesion. A similar result was achieved with dsAAV7 vector encoding the cerebral dopamine neurotrophic factor (CDNF), a protein with therapeutic potential. Viral vector-mediated intracerebral gene delivery has been used before in rodent models of ischemic injury. However, the method of targeting gene expression to the peri-infarct region, after the initial phase of ischemic cell death, has not been described before. We demonstrate a straightforward and robust way to target AAV vector-mediated over-expression of genes to the peri-infarct region in a rat stroke model. This method will be useful for studying the action of specific proteins in peri-infarct region during the recovery process. Copyright © 2014 Elsevier B.V. All rights reserved.

Effects of down-regulating ornithine decarboxylase upon putrescine-associated metabolism and growth in Nicotiana tabacum L.

PubMed Central

Dalton, Heidi L.; Blomstedt, Cecilia K.; Neale, Alan D.; Gleadow, Ros; DeBoer, Kathleen D.; Hamill, John D.

2016-01-01

Transgenic plants of Nicotiana tabacum L. homozygous for an RNAi construct designed to silence ornithine decarboxylase (ODC) had significantly lower concentrations of nicotine and nornicotine, but significantly higher concentrations of anatabine, compared with vector-only controls. Silencing of ODC also led to significantly reduced concentrations of polyamines (putrescine, spermidine and spermine), tyramine and phenolamides (caffeoylputrescine and dicaffeoylspermidine) with concomitant increases in concentrations of amino acids ornithine, arginine, aspartate, glutamate and glutamine. Root transcript levels of S-adenosyl methionine decarboxylase, S-adenosyl methionine synthase and spermidine synthase (polyamine synthesis enzymes) were reduced compared with vector controls, whilst transcript levels of arginine decarboxylase (putrescine synthesis), putrescine methyltransferase (nicotine production) and multi-drug and toxic compound extrusion (alkaloid transport) proteins were elevated. In contrast, expression of two other key proteins required for alkaloid synthesis, quinolinic acid phosphoribosyltransferase (nicotinic acid production) and a PIP-family oxidoreductase (nicotinic acid condensation reactions), were diminished in roots of odc-RNAi plants relative to vector-only controls. Transcriptional and biochemical differences associated with polyamine and alkaloid metabolism were exacerbated in odc-RNAi plants in response to different forms of shoot damage. In general, apex removal had a greater effect than leaf wounding alone, with a combination of these injury treatments producing synergistic responses in some cases. Reduced expression of ODC appeared to have negative effects upon plant growth and vigour with some leaves of odc-RNAi lines being brittle and bleached compared with vector-only controls. Together, results of this study demonstrate that ornithine decarboxylase has important roles in facilitating both primary and secondary metabolism in Nicotiana. PMID:27126795

Simulated Transmission of the Dengue Virus Across the US-Mexico Border Using Remotely Sensed and Ground Based Weather Data

NASA Technical Reports Server (NTRS)

Morin, Cory; Quattrochi, Dale A.

2015-01-01

Incidence of dengue fever, caused by a mosquito transmitted virus, have increased in the Americas during recent decades. In the US, local transmission has been reported in southern Texas and Florida. However, despite its close proximity to dengue endemic areas in Mexico and the presence of a primary mosquito vector, there are no reports of local transmission in Arizona. Many studies have demonstrated that weather influences dengue virus transmission by regulating vector development rates, vector habitat availability, and the duration of the virus extrinsic incubation period (EIP). The EIP, the period between mosquito infection and the ability for it to retransmit the virus, is especially important given its high sensitivity to temperature and the short lifespan of mosquitoes. Other studies, however, have suggested that human related factors such as socioeconomic status and herd immunity may explain much of the disparity in dengue incidence in the US-Mexico border region. Using a meteorologically driven model of vector population dynamics and virus transmission we compare simulations of dengue fever cases in southern Arizona and northern Mexico. A Monte Carlo approach is employed to select parameter values by evaluating simulations in Hermosillo Mexico with reported dengue fever case data. Simulations that replicate the case data best are retained and rerun using remotely sensed climate data from other Arizona and Mexico locations to determine the relative influence of weather on virus transmission. Although human and environmental factors undoubtedly influence dengue transmission in the US-Mexico border regions, weather is a major facilitator of the transmission process.

Modified Vaccinia Virus Ankara: History, Value in Basic Research, and Current Perspectives for Vaccine Development.

PubMed

Volz, A; Sutter, G

2017-01-01

Safety tested Modified Vaccinia virus Ankara (MVA) is licensed as third-generation vaccine against smallpox and serves as a potent vector system for development of new candidate vaccines against infectious diseases and cancer. Historically, MVA was developed by serial tissue culture passage in primary chicken cells of vaccinia virus strain Ankara, and clinically used to avoid the undesirable side effects of conventional smallpox vaccination. Adapted to growth in avian cells MVA lost the ability to replicate in mammalian hosts and lacks many of the genes orthopoxviruses use to conquer their host (cell) environment. As a biologically well-characterized mutant virus, MVA facilitates fundamental research to elucidate the functions of poxvirus host-interaction factors. As extremely safe viral vectors MVA vaccines have been found immunogenic and protective in various preclinical infection models. Multiple recombinant MVA currently undergo clinical testing for vaccination against human immunodeficiency viruses, Mycobacterium tuberculosis or Plasmodium falciparum. The versatility of the MVA vector vaccine platform is readily demonstrated by the swift development of experimental vaccines for immunization against emerging infections such as the Middle East Respiratory Syndrome. Recent advances include promising results from the clinical testing of recombinant MVA-producing antigens of highly pathogenic avian influenza virus H5N1 or Ebola virus. This review summarizes our current knowledge about MVA as a unique strain of vaccinia virus, and discusses the prospects of exploiting this virus as research tool in poxvirus biology or as safe viral vector vaccine to challenge existing and future bottlenecks in vaccinology. © 2017 Elsevier Inc. All rights reserved.

Hyaluronic acid enhances gene delivery into the cochlea.

PubMed

Shibata, Seiji B; Cortez, Sarah R; Wiler, James A; Swiderski, Donald L; Raphael, Yehoash

2012-03-01

Cochlear gene therapy can be a new avenue for the treatment of severe hearing loss by inducing regeneration or phenotypic rescue. One necessary step to establish this therapy is the development of a safe and feasible inoculation surgery, ideally without drilling the bony cochlear wall. The round window membrane (RWM) is accessible in the middle-ear space, but viral vectors placed on this membrane do not readily cross the membrane to the cochlear tissues. In an attempt to enhance permeability of the RWM, we applied hyaluronic acid (HA), a nontoxic and biodegradable reagent, onto the RWM of guinea pigs, prior to delivering an adenovirus carrying enhanced green fluorescent protein (eGFP) reporter gene (Ad-eGFP) at the same site. We examined distribution of eGFP in the cochlea 1 week after treatment, comparing delivery of the vector via the RWM, with or without HA, to delivery by a cochleostomy into the perilymph. We found that cochlear tissue treated with HA-assisted delivery of Ad-eGFP demonstrated wider expression of transgenes in cochlear cells than did tissue treated by cochleostomy injection. HA-assisted vector delivery facilitated expression in cells lining the scala media, which are less accessible and not transduced after perilymphatic injection. We assessed auditory function by measuring auditory brainstem responses and determined that thresholds were significantly better in the ears treated with HA-assisted Ad-eGFP placement on the RWM as compared with cochleostomy. Together, these data demonstrate that HA-assisted delivery of viral vectors provides an atraumatic and clinically feasible method to introduce transgenes into cochlear cells, thereby enhancing both research methods and future clinical application.

Bad-good constraints on a polarity correspondence account for the spatial-numerical association of response codes (SNARC) and markedness association of response codes (MARC) effects.

PubMed

Leth-Steensen, Craig; Citta, Richie

2016-01-01

Performance in numerical classification tasks involving either parity or magnitude judgements is quicker when small numbers are mapped onto a left-sided response and large numbers onto a right-sided response than for the opposite mapping (i.e., the spatial-numerical association of response codes or SNARC effect). Recent research by Gevers et al. [Gevers, W., Santens, S., Dhooge, E., Chen, Q., Van den Bossche, L., Fias, W., & Verguts, T. (2010). Verbal-spatial and visuospatial coding of number-space interactions. Journal of Experimental Psychology: General, 139, 180-190] suggests that this effect also arises for vocal "left" and "right" responding, indicating that verbal-spatial coding has a role to play in determining it. Another presumably verbal-based, spatial-numerical mapping phenomenon is the linguistic markedness association of response codes (MARC) effect whereby responding in parity tasks is quicker when odd numbers are mapped onto left-sided responses and even numbers onto right-sided responses. A recent account of both the SNARC and MARC effects is based on the polarity correspondence principle [Proctor, R. W., & Cho, Y. S. (2006). Polarity correspondence: A general principle for performance of speeded binary classification tasks. Psychological Bulletin, 132, 416-442]. This account assumes that stimulus and response alternatives are coded along any number of dimensions in terms of - and + polarities with quicker responding when the polarity codes for the stimulus and the response correspond. In the present study, even-odd parity judgements were made using either "left" and "right" or "bad" and "good" vocal responses. Results indicated that a SNARC effect was indeed present for the former type of vocal responding, providing further evidence for the sufficiency of the verbal-spatial coding account for this effect. However, the decided lack of an analogous SNARC-like effect in the results for the latter type of vocal responding provides an important constraint on the presumed generality of the polarity correspondence account. On the other hand, the presence of robust MARC effects for "bad" and "good" but not "left" and "right" vocal responses is consistent with the view that such effects are due to conceptual associations between semantic codes for odd-even and bad-good (but not necessarily left-right).

A Plasmodium falciparum strain expressing GFP throughout the parasite's life-cycle.

PubMed

Talman, Arthur M; Blagborough, Andrew M; Sinden, Robert E

2010-02-10

The human malaria parasite Plasmodium falciparum is responsible for the majority of malaria-related deaths. Tools allowing the study of the basic biology of P. falciparum throughout the life cycle are critical to the development of new strategies to target the parasite within both human and mosquito hosts. We here present 3D7HT-GFP, a strain of P. falciparum constitutively expressing the Green Fluorescent Protein (GFP) throughout the life cycle, which has retained its capacity to complete sporogonic development. The GFP expressing cassette was inserted in the Pf47 locus. Using this transgenic strain, parasite tracking and population dynamics studies in mosquito stages and exo-erythrocytic schizogony is greatly facilitated. The development of 3D7HT-GFP will permit a deeper understanding of the biology of parasite-host vector interactions, and facilitate the development of high-throughput malaria transmission assays and thus aid development of new intervention strategies against both parasite and mosquito.

Super Normal Vector for Human Activity Recognition with Depth Cameras.

PubMed

Yang, Xiaodong; Tian, YingLi

2017-05-01

The advent of cost-effectiveness and easy-operation depth cameras has facilitated a variety of visual recognition tasks including human activity recognition. This paper presents a novel framework for recognizing human activities from video sequences captured by depth cameras. We extend the surface normal to polynormal by assembling local neighboring hypersurface normals from a depth sequence to jointly characterize local motion and shape information. We then propose a general scheme of super normal vector (SNV) to aggregate the low-level polynormals into a discriminative representation, which can be viewed as a simplified version of the Fisher kernel representation. In order to globally capture the spatial layout and temporal order, an adaptive spatio-temporal pyramid is introduced to subdivide a depth video into a set of space-time cells. In the extensive experiments, the proposed approach achieves superior performance to the state-of-the-art methods on the four public benchmark datasets, i.e., MSRAction3D, MSRDailyActivity3D, MSRGesture3D, and MSRActionPairs3D.

Tandem SUMO fusion vectors for improving soluble protein expression and purification.

PubMed

Guerrero, Fernando; Ciragan, Annika; Iwaï, Hideo

2015-12-01

Availability of highly purified proteins in quantity is crucial for detailed biochemical and structural investigations. Fusion tags are versatile tools to facilitate efficient protein purification and to improve soluble overexpression of proteins. Various purification and fusion tags have been widely used for overexpression in Escherichia coli. However, these tags might interfere with biological functions and/or structural investigations of the protein of interest. Therefore, an additional purification step to remove fusion tags by proteolytic digestion might be required. Here, we describe a set of new vectors in which yeast SUMO (SMT3) was used as the highly specific recognition sequence of ubiquitin-like protease 1, together with other commonly used solubility enhancing proteins, such as glutathione S-transferase, maltose binding protein, thioredoxin and trigger factor for optimizing soluble expression of protein of interest. This tandem SUMO (T-SUMO) fusion system was tested for soluble expression of the C-terminal domain of TonB from different organisms and for the antiviral protein scytovirin. Copyright © 2015 Elsevier Inc. All rights reserved.

Implant Composed of Demineralized Bone and Mesenchymal Stem Cells Genetically Modified with AdBMP2/AdBMP7 for the Regeneration of Bone Fractures in Ovis aries

PubMed Central

Hernandez-Hurtado, Adelina A.; Lara-Arias, Jorge; Romero-Diaz, Viktor J.; Abrego-Guerra, Adalberto; Vilchez-Cavazos, Jose F.; Elizondo-Riojas, Guillermo; Martinez-Rodriguez, Herminia G.; Espinoza-Juarez, Marcela A.; Mendoza Lemus, Oscar F.

2016-01-01

Adipose-derived mesenchymal stem cells (ADMSCs) are inducible to an osteogenic phenotype by the bone morphogenetic proteins (BMPs). This facilitates the generation of implants for bone tissue regeneration. This study evaluated the in vitro osteogenic differentiation of ADMSCs transduced individually and in combination with adenoviral vectors expressing BMP2 and BMP7. Moreover, the effectiveness of the implant containing ADMSCs transduced with the adenoviral vectors AdBMP2/AdBMP7 and embedded in demineralized bone matrix (DBM) was tested in a model of tibial fracture in sheep. This graft was compared to ewes implanted with untransduced ADMSCs embedded in the same matrix and with injured but untreated animals. In vivo results showed accelerated osteogenesis in the group treated with the AdBMP2/AdBMP7 transduced ADMSC graft, which also showed improved restoration of the normal bone morphology. PMID:27818692

Recombinant soluble adenovirus receptor

DOEpatents

Freimuth, Paul I.

2002-01-01

Disclosed are isolated polypeptides from human CAR (coxsackievirus and adenovirus receptor) protein which bind adenovirus. Specifically disclosed are amino acid sequences which corresponds to adenovirus binding domain D1 and the entire extracellular domain of human CAR protein comprising D1 and D2. In other aspects, the disclosure relates to nucleic acid sequences encoding these domains as well as expression vectors which encode the domains and bacterial cells containing such vectors. Also disclosed is an isolated fusion protein comprised of the D1 polypeptide sequence fused to a polypeptide sequence which facilitates folding of D1 into a functional, soluble domain when expressed in bacteria. The functional D1 domain finds application for example in a therapeutic method for treating a patient infected with a virus which binds to D1, and also in a method for identifying an antiviral compound which interferes with viral attachment. Also included is a method for specifically targeting a cell for infection by a virus which binds to D1.

The Use of Synthetic Carriers in Malaria Vaccine Design

PubMed Central

Powles, Liam; Xiang, Sue D.; Selomulya, Cordelia; Plebanski, Magdalena

2015-01-01

Malaria vaccine research has been ongoing since the 1980s with limited success. However, recent improvements in our understanding of the immune responses required to combat each stage of infection will allow for intelligent design of both antigens and their associated delivery vaccine vehicles/vectors. Synthetic carriers (also known as vectors) are usually particulate and have multiple properties, which can be varied to control how an associated vaccine interacts with the host, and consequently how the immune response develops. This review comprehensively analyzes both historical and recent studies in which synthetic carriers are used to deliver malaria vaccines. Furthermore, the requirements for a synthetic carrier, such as size, charge, and surface chemistry are reviewed in order to understand the design of effective particle-based vaccines against malaria, as well as providing general insights. Synthetic carriers have the ability to alter and direct the immune response, and a better control of particle properties will facilitate improved vaccine design in the near future. PMID:26529028

The semantic representation of prejudice and stereotypes.

PubMed

Bhatia, Sudeep

2017-07-01

We use a theory of semantic representation to study prejudice and stereotyping. Particularly, we consider large datasets of newspaper articles published in the United States, and apply latent semantic analysis (LSA), a prominent model of human semantic memory, to these datasets to learn representations for common male and female, White, African American, and Latino names. LSA performs a singular value decomposition on word distribution statistics in order to recover word vector representations, and we find that our recovered representations display the types of biases observed in human participants using tasks such as the implicit association test. Importantly, these biases are strongest for vector representations with moderate dimensionality, and weaken or disappear for representations with very high or very low dimensionality. Moderate dimensional LSA models are also the best at learning race, ethnicity, and gender-based categories, suggesting that social category knowledge, acquired through dimensionality reduction on word distribution statistics, can facilitate prejudiced and stereotyped associations. Copyright © 2017 Elsevier B.V. All rights reserved.

A survey of mosquitoes breeding in used tires in Spain for the detection of imported potential vector species.

PubMed

Roiz, D; Eritja, R; Escosa, R; Lucientes, J; Marquès, E; Melero-Alcíbar, R; Ruiz, S; Molina, R

2007-06-01

The used tire trade has facilitated the introduction, spread, and establishment of the Asian tiger mosquito, Aedes albopictus, and other mosquito species in several countries of America, Africa, Oceania, and Europe. A strategy for detecting these imported mosquito vectors was developed in Spain during 2003-2004 by EVITAR (multidisciplinary network for the study of viruses transmitted by arthropods and rodents). A survey in 45 locations found no invasive species. Eight autochthonous species of mosquitoes were detected in used tires, including Culex pipiens, Cx. hortensis, Cx. modestus, Anopheles atroparvus, An. claviger, Culiseta longiareolata, Cs. annulata, and Aedes caspius. Dominant species were Cx. pipiens and Cs. longiareolata. Aedes caspius was found in only once, near its natural breeding habitat. Considering the recent discovery of an established population of Ae. albopictus in Catalonia, the increasing commerce of used tires in Spain for recycling, storage, and recapping might greatly contribute to the rapid spread of this species across the Iberian Peninsula.

The national assessment of shoreline change: A GIS compilation of vector shorelines and associated shoreline change data for the New England and Mid-Atlantic Coasts

USGS Publications Warehouse

Himmelstoss, Emily A.; Kratzmann, Meredith G.; Hapke, Cheryl; Thieler, E. Robert; List, Jeffrey

2010-01-01

Sandy ocean beaches are a popular recreational destination, often surrounded by communities containing valuable real estate. Development is on the rise despite the fact that coastal infrastructure is subjected to flooding and erosion. As a result, there is an increased demand for accurate information regarding past and present shoreline changes. The U.S. Geological Survey's National Assessment of Shoreline Change Project has compiled a comprehensive database of digital vector shorelines and shoreline-change rates for the New England and Mid-Atlantic Coasts. There is currently no widely accepted standard for analyzing shoreline change. Existing measurement and rate-calculation methods vary from study to study and preclude combining results into statewide or regional assessments. The impetus behind the National Assessment project was to develop a standardized method that is consistent from coast to coast for measuring changes in shoreline position. The goal was to facilitate the process of periodically and systematically updating the results in an internally consistent manner.

Nucleic acid sequences encoding D1 and D1/D2 domains of human coxsackievirus and adenovirus receptor (CAR)

DOEpatents

Freimuth, Paul I.

2010-04-06

The invention provides recombinant human CAR (coxsackievirus and adenovirus receptor) polypeptides which bind adenovirus. Specifically, polypeptides corresponding to adenovirus binding domain D1 and the entire extracellular domain of human CAR protein comprising D1 and D2 are provided. In another aspect, the invention provides nucleic acid sequences encoding these domains and expression vectors for producing the domains and bacterial cells containing such vectors. The invention also includes an isolated fusion protein comprised of the D1 polypeptide fused to a polypeptide which facilitates folding of D1 when expressed in bacteria. The functional D1 domain finds application in a therapeutic method for treating a patient infected with a CAR D1-binding virus, and also in a method for identifying an antiviral compound which interferes with viral attachment. The invention also provides a method for specifically targeting a cell for infection by a virus which binds to D1.

Silencing urease: a key evolutionary step that facilitated the adaptation of Yersinia pestis to the flea-borne transmission route.

PubMed

Chouikha, Iman; Hinnebusch, B Joseph

2014-12-30

The arthropod-borne transmission route of Yersinia pestis, the bacterial agent of plague, is a recent evolutionary adaptation. Yersinia pseudotuberculosis, the closely related food-and water-borne enteric species from which Y. pestis diverged less than 6,400 y ago, exhibits significant oral toxicity to the flea vectors of plague, whereas Y. pestis does not. In this study, we identify the Yersinia urease enzyme as the responsible oral toxin. All Y. pestis strains, including those phylogenetically closest to the Y. pseudotuberculosis progenitor, contain a mutated ureD allele that eliminated urease activity. Restoration of a functional ureD was sufficient to make Y. pestis orally toxic to fleas. Conversely, deletion of the urease operon in Y. pseudotuberculosis rendered it nontoxic. Enzymatic activity was required for toxicity. Because urease-related mortality eliminates 30-40% of infective flea vectors, ureD mutation early in the evolution of Y. pestis was likely subject to strong positive selection because it significantly increased transmission potential.

Silencing urease: A key evolutionary step that facilitated the adaptation of Yersinia pestis to the flea-borne transmission route

PubMed Central

Chouikha, Iman; Hinnebusch, B. Joseph

2014-01-01

The arthropod-borne transmission route of Yersinia pestis, the bacterial agent of plague, is a recent evolutionary adaptation. Yersinia pseudotuberculosis, the closely related food-and water-borne enteric species from which Y. pestis diverged less than 6,400 y ago, exhibits significant oral toxicity to the flea vectors of plague, whereas Y. pestis does not. In this study, we identify the Yersinia urease enzyme as the responsible oral toxin. All Y. pestis strains, including those phylogenetically closest to the Y. pseudotuberculosis progenitor, contain a mutated ureD allele that eliminated urease activity. Restoration of a functional ureD was sufficient to make Y. pestis orally toxic to fleas. Conversely, deletion of the urease operon in Y. pseudotuberculosis rendered it nontoxic. Enzymatic activity was required for toxicity. Because urease-related mortality eliminates 30–40% of infective flea vectors, ureD mutation early in the evolution of Y. pestis was likely subject to strong positive selection because it significantly increased transmission potential. PMID:25453069

A fusion of minicircle DNA and nanoparticle delivery technologies facilitates therapeutic genetic engineering of autologous canine olfactory mucosal cells.

PubMed

Delaney, Alexander M; Adams, Christopher F; Fernandes, Alinda R; Al-Shakli, Arwa F; Sen, Jon; Carwardine, Darren R; Granger, Nicolas; Chari, Divya M

2017-06-29

Olfactory ensheathing cells (OECs) promote axonal regeneration and improve locomotor function when transplanted into the injured spinal cord. A recent clinical trial demonstrated improved motor function in domestic dogs with spinal injury following autologous OEC transplantation. Their utility in canines offers promise for human translation, as dogs are comparable to humans in terms of clinical management and genetic/environmental variation. Moreover, the autologous, minimally invasive derivation of OECs makes them viable for human spinal injury investigation. Genetic engineering of transplant populations may augment their therapeutic potential, but relies heavily on viral methods which have several drawbacks for clinical translation. We present here the first proof that magnetic particles deployed with applied magnetic fields and advanced DNA minicircle vectors can safely bioengineer OECs to secrete a key neurotrophic factor, with an efficiency approaching that of viral vectors. We suggest that our alternative approach offers high translational potential for the delivery of augmented clinical cell therapies.

A new technique for rapid assessment of eutrophication status of coastal waters using a support vector machine

NASA Astrophysics Data System (ADS)

Kong, Xianyu; Che, Xiaowei; Su, Rongguo; Zhang, Chuansong; Yao, Qingzhen; Shi, Xiaoyong

2017-05-01

There is an urgent need to develop efficient evaluation tools that use easily measured variables to make rapid and timely eutrophication assessments, which are important for marine health management, and to implement eutrophication monitoring programs. In this study, an approach for rapidly assessing the eutrophication status of coastal waters with three easily measured parameters (turbidity, chlorophyll a and dissolved oxygen) was developed by the grid search (GS) optimized support vector machine (SVM), with trophic index TRIX classification results as the reference. With the optimized penalty parameter C =64 and the kernel parameter γ =1, the classification accuracy rates reached 89.3% for the training data, 88.3% for the cross-validation, and 88.5% for the validation dataset. Because the developed approach only used three easy-to-measure variables, its application could facilitate the rapid assessment of the eutrophication status of coastal waters, resulting in potential cost savings in marine monitoring programs and assisting in the provision of timely advice for marine management.

Dual Insect specific virus infection limits Arbovirus replication in Aedes mosquito cells.

PubMed

Schultz, Michaela J; Frydman, Horacio M; Connor, John H

2018-05-01

Aedes mosquitoes are vectors for many pathogenic viruses. Cell culture systems facilitate the investigation of virus growth in the mosquito vector. We found Zika virus (ZIKV) growth to be consistent in A. albopictus cells but hypervariable in A. aegypti cell lines. As a potential explanation of this variability, we tested the hypothesis that our cells harbored opportunistic viruses. We screened Aedes cell lines for the presence of insect specific viruses (ISVs), Cell-fusing agent virus (CFAV) and Phasi charoen-like virus (PCLV). PCLV was present in the ZIKV-growth-variable A. aegypti cell lines but absent in A. albopictus lines, suggesting that these ISVs may interfere with ZIKV growth. In support of this hypothesis, PCLV infection of CFAV-positive A. albopictus cells inhibited the growth of ZIKV, dengue virus and La Crosse virus. These data suggest ISV infection of cell lines can impact arbovirus growth leading to significant changes in cell permissivity to arbovirus infection. Copyright © 2018 Elsevier Inc. All rights reserved.

Live Virus Vaccines Based on a Yellow Fever Vaccine Backbone: Standardized Template with Key Considerations for a Risk/Benefit Assessment*

PubMed Central

Monath, Thomas P.; Seligman, Stephen J.; Robertson, James S.; Guy, Bruno; Hayes, Edward B.; Condit, Richard C.; Excler, Jean Louis; Mac, Lisa Marie; Carbery, Baevin; Chen, Robert T

2015-01-01

The Brighton Collaboration Viral Vector Vaccines Safety Working Group (V3SWG) was formed to evaluate the safety of live, recombinant viral vaccines incorporating genes from heterologous viruses inserted into the backbone of another virus (so-called “chimeric virus vaccines”). Many viral vector vaccines are in advanced clinical trials. The first such vaccine to be approved for marketing (to date in Australia, Thailand, Malaysia, and the Philippines) is a vaccine against the flavivirus Japanese encephalitis (JE), which employs a licensed vaccine (yellow fever 17D) as a vector. In this vaccine, two envelope proteins (prM-E) of YF 17D virus were replaced by the corresponding genes of JE virus, with additional attenuating mutations incorporated into the JE gene inserts. Similar vaccines have been constructed by inserting prM-E genes of dengue and West Nile into YF 17D virus and are in late stage clinical studies. The dengue vaccine is, however, more complex in that it requires a mixture of four live vectors each expressing one of the four dengue serotypes. This vaccine has been evaluated in multiple clinical trials. No significant safety concerns have been found. The Phase 3 trials met their endpoints in terms of overall reduction of confirmed dengue fever, and, most importantly a significant reduction in severe dengue and hospitalization due to dengue. However, based on results that have been published so far, efficacy in preventing serotype 2 infection is less than that for the other three serotypes. In the development of these chimeric vaccines, an important series of comparative studies of safety and efficacy were made using the parental YF 17D vaccine virus as a benchmark. In this paper, we use a standardized template describing the key characteristics of the novel flavivirus vaccine vectors, in comparison to the parental YF 17D vaccine. The template facilitates scientific discourse among key stakeholders by increasing the transparency and comparability of information. The Brighton Collaboration V3SWG template may also be useful as a guide to the evaluation of other recombinant viral vector vaccines. PMID:25446819

Attraction of the cutaneous leishmaniasis vector Nyssomyia neivai (Diptera: Psychodidae) to host odour components in a wind tunnel

PubMed Central

2012-01-01

Background Laboratory studies of host-seeking olfactory behaviour in sandflies have largely been restricted to the American visceral leishmaniasis vector Lutzomyia longipalpis. In comparison, almost nothing is known about the chemical ecology of related species, which transmit American cutaneous leishmaniasis (ACL), due in part to difficulties in raising these insects in the laboratory. Understanding how ACL vectors locate their hosts will be essential to developing new vector control strategies to combat this debilitating disease. Methods This study examined host-odour seeking behaviour of the ACL vector Nyssomyia neivai (Pinto) (=Lutzomyia neivai) using a wind tunnel olfactometer. The primary aim was to determine whether field-collected female N. neivai would respond to host odours in the laboratory, thereby eliminating the need to maintain colonies of these insects for behavioural experiments. Responses to two key host odour components, 1-octen-3-ol and lactic acid, and a commercially-available mosquito lure (BG-Lure™) were assessed and compared relative to an air control. We also tested whether trials could be conducted outside of the normal evening activity period of N. neivai without impacting on fly behaviour, and whether the same flies could be used to assess baseline responses to air without affecting responses to octenol, thereby reducing the number of flies required for experiments. Results Octenol was found to both activate host-seeking behaviour and attract female N. neivai in the wind tunnel, while lactic acid elicited weaker responses of activation and attractiveness under identical conditions. The BG-Lure did not activate or attract N. neivai under test conditions. Further experiments showed that sandfly behaviour in the wind tunnel was not affected by time of day, such that experiments need not be restricted to nocturnal hours. Moreover, using the same flies to measure both baseline responses to air and attraction to test compounds did not affect odour-seeking behaviour. Conclusions The results of this study demonstrate that N. neivai taken from the field are suitable for use in laboratory olfactometer experiments. It is hoped this work will facilitate further research into chemical ecology of this species, and other ACL vectors. PMID:23009099

Attraction of the cutaneous leishmaniasis vector Nyssomyia neivai (Diptera: Psychodidae) to host odour components in a wind tunnel.

PubMed

Pinto, Mara C; Bray, Daniel P; Eiras, Alvaro E; Carvalheira, Henrique P; Puertas, Camila P

2012-09-25

Laboratory studies of host-seeking olfactory behaviour in sandflies have largely been restricted to the American visceral leishmaniasis vector Lutzomyia longipalpis. In comparison, almost nothing is known about the chemical ecology of related species, which transmit American cutaneous leishmaniasis (ACL), due in part to difficulties in raising these insects in the laboratory. Understanding how ACL vectors locate their hosts will be essential to developing new vector control strategies to combat this debilitating disease. This study examined host-odour seeking behaviour of the ACL vector Nyssomyia neivai (Pinto) (=Lutzomyia neivai) using a wind tunnel olfactometer. The primary aim was to determine whether field-collected female N. neivai would respond to host odours in the laboratory, thereby eliminating the need to maintain colonies of these insects for behavioural experiments. Responses to two key host odour components, 1-octen-3-ol and lactic acid, and a commercially-available mosquito lure (BG-Lure™) were assessed and compared relative to an air control. We also tested whether trials could be conducted outside of the normal evening activity period of N. neivai without impacting on fly behaviour, and whether the same flies could be used to assess baseline responses to air without affecting responses to octenol, thereby reducing the number of flies required for experiments. Octenol was found to both activate host-seeking behaviour and attract female N. neivai in the wind tunnel, while lactic acid elicited weaker responses of activation and attractiveness under identical conditions. The BG-Lure did not activate or attract N. neivai under test conditions. Further experiments showed that sandfly behaviour in the wind tunnel was not affected by time of day, such that experiments need not be restricted to nocturnal hours. Moreover, using the same flies to measure both baseline responses to air and attraction to test compounds did not affect odour-seeking behaviour. The results of this study demonstrate that N. neivai taken from the field are suitable for use in laboratory olfactometer experiments. It is hoped this work will facilitate further research into chemical ecology of this species, and other ACL vectors.

Live virus vaccines based on a yellow fever vaccine backbone: standardized template with key considerations for a risk/benefit assessment.

PubMed

Monath, Thomas P; Seligman, Stephen J; Robertson, James S; Guy, Bruno; Hayes, Edward B; Condit, Richard C; Excler, Jean Louis; Mac, Lisa Marie; Carbery, Baevin; Chen, Robert T

2015-01-01

The Brighton Collaboration Viral Vector Vaccines Safety Working Group (V3SWG) was formed to evaluate the safety of live, recombinant viral vaccines incorporating genes from heterologous viruses inserted into the backbone of another virus (so-called "chimeric virus vaccines"). Many viral vector vaccines are in advanced clinical trials. The first such vaccine to be approved for marketing (to date in Australia, Thailand, Malaysia, and the Philippines) is a vaccine against the flavivirus, Japanese encephalitis (JE), which employs a licensed vaccine (yellow fever 17D) as a vector. In this vaccine, two envelope proteins (prM-E) of YF 17D virus were exchanged for the corresponding genes of JE virus, with additional attenuating mutations incorporated into the JE gene inserts. Similar vaccines have been constructed by inserting prM-E genes of dengue and West Nile into YF 17D virus and are in late stage clinical studies. The dengue vaccine is, however, more complex in that it requires a mixture of four live vectors each expressing one of the four dengue serotypes. This vaccine has been evaluated in multiple clinical trials. No significant safety concerns have been found. The Phase 3 trials met their endpoints in terms of overall reduction of confirmed dengue fever, and, most importantly a significant reduction in severe dengue and hospitalization due to dengue. However, based on results that have been published so far, efficacy in preventing serotype 2 infection is less than that for the other three serotypes. In the development of these chimeric vaccines, an important series of comparative studies of safety and efficacy were made using the parental YF 17D vaccine virus as a benchmark. In this paper, we use a standardized template describing the key characteristics of the novel flavivirus vaccine vectors, in comparison to the parental YF 17D vaccine. The template facilitates scientific discourse among key stakeholders by increasing the transparency and comparability of information. The Brighton Collaboration V3SWG template may also be useful as a guide to the evaluation of other recombinant viral vector vaccines. Copyright © 2014. Published by Elsevier Ltd.

Transmission of Rift Valley fever virus from European-breed lambs to Culex pipiens mosquitoes.

PubMed

Vloet, Rianka P M; Vogels, Chantal B F; Koenraadt, Constantianus J M; Pijlman, Gorben P; Eiden, Martin; Gonzales, Jose L; van Keulen, Lucien J M; Wichgers Schreur, Paul J; Kortekaas, Jeroen

2017-12-01

Rift Valley fever virus (RVFV) is a mosquito-borne bunyavirus of the genus Phlebovirus that is highly pathogenic to ruminants and humans. The disease is currently confined to Africa and the Arabian Peninsula, but globalization and climate change may facilitate introductions of the virus into currently unaffected areas via infected animals or mosquitoes. The consequences of such an introduction will depend on environmental factors, the availability of susceptible ruminants and the capacity of local mosquitoes to transmit the virus. We have previously demonstrated that lambs native to the Netherlands are highly susceptible to RVFV and we here report the vector competence of Culex (Cx.) pipiens, the most abundant and widespread mosquito species in the country. Vector competence was first determined after artificial blood feeding of laboratory-reared mosquitoes using the attenuated Clone 13 strain. Subsequently, experiments with wild-type RVFV and mosquitoes hatched from field-collected eggs were performed. Finally, the transmission of RVFV from viremic lambs to mosquitoes was studied. Artificial feeding experiments using Clone 13 demonstrated that indigenous, laboratory-reared Cx. pipiens mosquitoes are susceptible to RVFV and that the virus can be transmitted via their saliva. Experiments with wild-type RVFV and mosquitoes hatched from field-collected eggs confirmed the vector competence of Cx. pipiens mosquitoes from the Netherlands. To subsequently investigate transmission of the virus under more natural conditions, mosquitoes were allowed to feed on RVFV-infected lambs during the viremic period. We found that RVFV is efficiently transmitted from lambs to mosquitoes, although transmission was restricted to peak viremia. Interestingly, in the mosquito-exposed skin samples, replication of RVFV was detected in previously unrecognized target cells. We here report the vector competence of Cx. pipiens mosquitoes from the Netherlands for RVFV. Both laboratory-reared mosquitoes and well as those hatched from field-collected eggs were found to be competent vectors. Moreover, RVFV was transmitted efficiently from indigenous lambs to mosquitoes, although the duration of host infectivity was found to be shorter than previously assumed. Interestingly, analysis of mosquito-exposed skin samples revealed previously unidentified target cells of the virus. Our findings underscore the value of including natural target species in vector competence experiments.

Contents of the JPL Distributed Active Archive Center (DAAC) archive, version 2-91

NASA Technical Reports Server (NTRS)

Smith, Elizabeth A. (Editor); Lassanyi, Ruby A. (Editor)

1991-01-01

The Distributed Active Archive Center (DAAC) archive at the Jet Propulsion Laboratory (JPL) includes satellite data sets for the ocean sciences and global change research to facilitate multidisciplinary use of satellite ocean data. Parameters include sea surface height, surface wind vector, sea surface temperature, atmospheric liquid water, and surface pigment concentration. The Jet Propulsion Laboratory DAAC is an element of the Earth Observing System Data and Information System (EOSDIS) and will be the United States distribution site for the Ocean Topography Experiment (TOPEX)/POSEIDON data and metadata.

JPL Physical Oceanography Distributed Active Archive Center (PO.DAAC) data availability, version 1-94

NASA Technical Reports Server (NTRS)

1994-01-01

The Physical Oceanography Distributed Active Archive Center (PO.DAAC) archive at the Jet Propulsion Laboratory (JPL) includes satellite data sets for the ocean sciences and global-change research to facilitate multidisciplinary use of satellite ocean data. Parameters include sea-surface height, surface-wind vector, sea-surface temperature, atmospheric liquid water, and integrated water vapor. The JPL PO.DAAC is an element of the Earth Observing System Data and Information System (EOSDIS) and is the United States distribution site for Ocean Topography Experiment (TOPEX)/POSEIDON data and metadata.

Human habitat positioning system for NASA's space flight environmental simulator

NASA Technical Reports Server (NTRS)

Caldwell, W. F.; Tucker, J.; Keas, P.

1998-01-01

Artificial gravity by centrifugation offers an effective countermeasure to the physiologic deconditioning of chronic exposure to microgravity; however, the system requirements of rotational velocity, radius of rotation, and resultant centrifugal acceleration require thorough investigation to ascertain the ideal human-use centrifuge configuration. NASA's Space Flight Environmental Simulator (SFES), a 16-meter (52-foot) diameter, animal-use centrifuge, was recently modified to accommodate human occupancy. This paper describes the SFES Human Habitat Positioning System, the mechanism that facilitates radius of rotation variability and alignment of the centrifuge occupants with the artificial gravity vector.

Implementation of Gravity Model to Estimation of Transportation Market Shares

NASA Astrophysics Data System (ADS)

Krata, Przemysław

2010-03-01

The theoretical consideration presented in the paper is inspired by market gravity models, as an interesting attitude towards operations research on a market. The transportation market issues are emphasized. The mathematical model of relations, taking place between transportation companies and their customers on the market, which is applied in the course of the research is based on continuous functions characteristics. This attitude enables the use of the field theory notions. The resultant vector-type utility function facilitates obtaining of competitive advantage areas for all transportation companies located on the considered transportation market.